MSC SabryAbdallah

TOXICOLOGICAL STUDIES OF SOME PESTICIDES IN RELATION TO

THEIR SIDE EFFECTS.

by

SABRY ABDEL-MONEM ABD-ELAAL ABD-ALLAH

B.Sc. Agric.,(Pesticides), Tanta Univ.,1991

THESIS

Submitted in partial fulfilment of the requirements for the degree of Master of Science

In

Department of Pesticides Faculty of Agriculture, Kafer El-Sheikh

Tanta University

(1998)

TOXICOLOGICAL STUDIES OF SOME PESTICIDES IN RELATION

TO THEIR SIDE EFFECTS.

by

Sabry Abdel-Monem Abd-El-Aal Abd-Allah

For the degree of

M. Sc. in Pesticides.

Examiners committee Approved by

1-Prof. Dr. Mohamed Aly Ashry.

Prof. of pesticides chemistry, and Vice Dean

of the Facaculty of Agriculure., Kafr El-

Sheikh, Tanta University.

---------------------------

2-Prof. Dr. Ahmed El-Sayed Salama.

Prof. of pesticides, and Chairman of pesticides

Dept. Faculty of Agriculure, Kafr El-Sheikh,

Tanta University.

---------------------------

3-Prof. Dr.Moustafa A. Abbassy.

Prof. of pesticides and Dean of the Faculty of

Agriculure., Damanhour, Alex. University.

---------------------------

4- Dr. Sherif E. El-Hamady.

Asso. Prof. of pesticides, Faculty of

Agriculure., Kafr El-Sheikh

Tanta University.

---------------------------

Date 8 / 3 /1998

Advisors Committee

1-Prof. Dr.Mohamed Aly Ashry.

Prof. of Pesticides, and Vice Dean of the Fac. of Agric., Kafr El-Sheikh,

Tanta University.

2-Prof. Dr. Mohamed A. Abd-Elbaki

Prof. of Pesticides, Fac. of Agric., Kafr El-Sheikh, Tanta University

3- Dr. Sherif E. El-Hamady.

Associate Prof. of Pesticides, Fac. of Agric., Kafr El-Sheikh, Tanta

University.

CONTENTS

CONTENTS

ACKNOWLEDGMENT.

INTRODUCTION. 1

REVIEW OF LITERATURE. 3

1-Pesticidal efficiency against pests and their natural enemies

especially of vegetable crops.

3

1.1. Cotton leafworm Spodoptera littoralis. 4

1.2. Aphids. 13

1.3. Whitefly, Bemisia tabaci. 29

1.4. Spider mites. 35

1.5. Beneficial arthropodes. 41

2- Mammalian toxicity of pesticides. 50

2.1. The effect on AChE and esterases. 51

2.2. The effect on liver function and other biochemical

parameters.

63

2.2.1. Transaminase. 64

2. 2. 2. Alkaline phosphatase 73

2. 2. 3 Cholesterol, bilirubin, total protien and albumin 78

2.3. Kidney function. 84

2.4. The effect on body weight. 86

2. 5. Histopathological studies. 91

MATERIALS AND METHODS. 95

1- Test insects. 95

1.1. The cotton leafworm: Spodoptera littoralis (Boisd). 95

1.2. Aphids. 95

1.3. Spider mite Tetranychus cinnabarinus (boisduval). 96

1.4. The predator, paederus alfierii (Kock). 97

2-Test animals. 97

I

CONTENTS

3-Pesticides. 97

4-Laboratory-toxicity tests. 101

4.1. The cotton leaf worm, Spodoptera littoralis. 101

4.2. Aphids. 101

4.3. Mites. 102

4.4. The predator, P. alfierii. 102

4.5. L.D.P lines and statistical analysis. 103

5-Field studies. 103

6- Acute toxicity to rats. 104

7-Daily oral dose treatment. 105

7.1. Experimental. 105

7.2. Serum preparation. 106

8- Biochemical analysis. 106

8.1. Determanation of total protien. 106

8.2. Determination of non-specific esterases activity. 107

8.3.Determination of acetylcholinestrase activity. 108

8.4.Determination of transaminases. 110

8.5.Determination of alkaline phosphatase. 112

8.6.Determination of cholesterol. 115

8.7.Determination of total bilirubin. 116

8.8.Detemination of Albumin. 117

8.9.Determination of creatinine. 118

8.10.Determination of uric acid. 119

9.Histopathological studies. 120

10-Statical analysis 123

RESULTS AND DISCUSSION. 124

1-Pesticidal activity of tested chemicals. 124

1.1. Laboratory tests. 124

II

CONTENTS

1.1.1. Toxicity to the cotton leafworm, S. littoralis. 124

1.1.2. Toxicity to aphids. 125

1.1.3 Toxicity to the spider mite T. cinnabarinns. 132

1.1.4. Toxicity to the predator Paederus alfirii. 133

1.2. Field studies. 137

1.2.1 The cotton leafworm, S. littoralis. 137

1.2.2 The whitefly, Bemisia tabaci. 142

1.2.3. Aphids, A. gossypii. 156

2-Toxicity of chloropyrifos-methyl and pirimicarb to white

rats.

161

2.1. Acute toxicity. 161

2.2. Sub-chronic toxicities. 164

2.2.1 Clinical symptoms and moralities through test period. 164

2.2.2. Effects on ChE and esterases specific activities. 165

2.2.3 The effect on liver function. 167

2.2.4 The effect on kidney function. 190

2.2.5 The effect on body weight gain. 192

2.2.6 Histopathological examination. 195

2.2.6.1 Histopathological effect in livers of treated rats. 195

2.2.6.2 Histopathological effect in kidneys of treated rats. 195

CONCLUSION. 203

ENGLISH SUMMARY. 204

REFERANCES. 209

ARABIC SUMMARY. -------

III

ACKNOWLEDGMENT

I strongly owe my thanks to ALLAH for lighting me the way and

directing me across every success.

The auther wishes to express his thank fulness to Dr. M. A. Ashry prof.

of chemistry of pesticides and Vice Dean of the Fac. of Agric., Tanta

Univ., for his supervision during the course of these studies and during

revision of the manuscript.

I wish to express my deep gratitude to Dr. Ahmed E. Salama Chairman

of the Chemistry of pesticides, Fac. of Agric., Tanta Univ., for his kind

help and advice throut this study.

Deepest and sincere gratitude to Dr. Mohamed A. Abd-Elbaki Prof.

Chemistry of pesticides. Fac. of Agric., Tanta Univ., for suggesting the

problem, supervising the work, desiging the experiment and providing

technical help and valuable scientific assistance.

The authour wishes also to thank Dr. Sherif E. El-Hamady. Associate

Prof. of pesticides, Fac. of Agric., Kafr El-Sheikh, Tanta University, for,

his valuable supervision, scientific suggestions, guidance, and kind help

during the period of the work, efforts in revising the manuscript and

discussing data deserve my gratitude and indebitedness.

Many thanks are also due to Dr. A. Rawash, Lecturer of Fac. of

Veterinary Medecine Kafr El-Sheikh, Tanta Univ., for his kind help in the

part of histopathological study.

Deep thank are also to all members in pesticides Dep., Fac. of Agric.,

Kafr El-Sheikh, Tanta Univ, for their continaous encouragement and

offering all facilities throught this work.

INTRODUCTIONINTRODUCTION

INTRODUCTION

The use of pesticides has enormously increased during the last 30

years in the world causing an obvious increase in the agricultural

production. Chemical control measures are usually effective, potentially

cover the control of most pests and diseases and may be used effectively

under a wide variety of conditions. However they can cause undesirable

non-target effects and leave undesirable residues (Oomen 1992). The

widely spread range of continuously developing insecticides has been

regularly reported as one of the most detrimental agents to human (Zidan

et al, 1996).

Indeed, chemical control measures are the least preferred of all

effective measures. Nevertheless, these measures and non-chemical ones

are very much complementary and there is no reason to expect a major

change in this situation (Oomen 1992).

Different organophosphate and carbamate insecticides are widely

appled in Egypt to control economic pests especially on vegetable for

their broad insecticidal spectrum and short persistence. However, certain

profiles of these pesticides are required to cover the needs of integrated

control particularly on vegetable crops. Pesticides chosen to be applied

should have both desired and undesired effects. Desired means effective

control or prevention of the pest whereas undesired are adverse effects on

beneficial organisms (e.g. natural enemies), on the consumers, (pesticides

residues on the product and their toxicological effects and on the workers

in crop growing (exposure to chemicals and their effects). Thus, the

availability of a number of different pesticides and reliable information

on their activity spectra and their negative side effects to beneficial

INTRODUCTION

arthropods and mammals is the subject of this thesis. Therefore the

following experiments were carried out:

1-Laboratory comparative studies for the toxicity of eight

organophosphate and carbamate insecticides to certain pests (i.e. sucking

pests, cotton leafworm and the beneficial predator, Paederus alfierii )

2- Field studies for evaluating the efficiency of the tested

insecticides against pests on some vegetable crops (i.e. Okra and squash).

3- The most efficient insecticides were subjected to mammalian

toxicological studies. In this respect, acute and subchronic toxicity

studies were performed on white rats.

2

REVIEWREVIEW OFOF

LITERATURELITERATURE

REVIEW OF LITERATURE

1-Pesticidal efficiency against pests and their

natural enemies especially of vegetable crops.

Vegetable crops (e.g. squash and okra) are subjected to severe

infestation with various arthropodal pests. Among of which are sucking

pests (e. g. whitefly, aphids and mites) and lepidopterus insects like

cotton leafworm, Spodoptera littoralis. In Egypt, the whitefly Bemisia

tabaci has been considered as an economic and serious pest which attacks

traditional or protected vegetable crops (Zidan et al. 1994). The whitefly

transmit plant virus that reduces the yield and quality of vegetables. Due

to the extensive application of pesticides against the whitefly, it has

become resistant to many groups of insecticides (Prodbhaker et al. 1985,

& El-Maghraby et al. 1997). Also, Aphis gossypii, A. craccivora and

Myzus persicae are among the most important and destructive pests that

attack several vegetable plants in Egypt. Economic importance of aphids

is attributable to their high reproductive potential and ability to transmit

plant pathogenic viruses (Mansour et al. 1977; Harris and

Maramorosch, 1977 and Darwish et al, 1994).

The phytophagous mites, Tetranychus sp (T. urticae and T.

cinnabarinus) attack the leaves of vegetables especially those of

Cucurbitaceae and as a result of infestation, plants are weakened and give

a very poor yield. Various lepidopteran insects (among of which is S.

littoralis) also attack vegetable plants leading to severe damage in the

foliage.

Continuous usage of several insecticides for controlling agricultural

arthropod pests has inevitably been followed by target pest resurgence,


secondary pest outbreaks, and the development of insecticide resistance

in target pests. All of these consequences are related to the disruption of

the natural enemies.

Hence, over the last 40 years, many investigators had annually

bioassayed hundreds of chemical insecticides belonging to different

groups against the perforated pests and also studied the discriminating of

it between pests and their predators , either under field or laboratory

conditions. Some of the above-mentioned studies are reviewed here:

1-1 Cotton leafworm, Spodoptera littoralis

Tantawy et al. (1974), studied the comparative toxicity using

topical application technique for formulated and technical insecticides,

Cyolane, Cytrolane, Nuvacron, and Dursban against Spodoptera littoralis

larvae. They indicated that formulated material are always more toxic

than the corresponding technical ones, and Dursban was the most toxic.

EL-Gayar et al. (1979), evaluated the initial and residual toxic

effect of single and mixture insecticides under field conditions against the

2nd and 4th instar larvae of S. littoralis. According to the average initial

mortality and average mortality during 9 days from spraying, the

insecticides can be grouped into the following groups : (1) Extremely

potent insecticides, i.e. Phosvel, Cyolane, Tamaron + Gusathion, Gardona

SP., and Cytrolane; (2) Highly potent insecticides, i.e. Tamaron, Orthene,

Elsan + EPN, Folaton, and Lannate; (3) Potent insecticides, i.e Gardona

W.P.; 4) Moderately potent insecticides, i.e. CRD; (5) Less potent

insecticides, i.e. Dipterex, Diazinon, Elsan, Actelic, Nuvacron, Galecron

SP, and Galecron EC. The first group showed highly residual effect and

was suggested for the control of S. littoralis in cotton fields. While

Lannate, Folaton, and Dipterex which showed highly initial effect and

4


low residual action were suggested for the control of the same pest in

vegetable plantations.

Watson and Guirguis. (1983), studied the efficiency of granular

insecticides against cotton pests. A granular formulations of aldicarb at

0.38 and 0.5, isazofos at 0.38 and 0.5, profenofos at 0.25-0.42, phosfolan

at 0.1-0.17, chlorpyrifos at 0.25-0.42 and chlorfenvinphos at 0.5-0.76 kg

a.i./ha were applied in the laboratory and field against pests of cotton, The

descending order of effectiveness of the compounds against larvae of S.

littoralis was phosfolan, chlorpyrifos, profenofos, isazofos, aldicarb and

chlorfenvinphos.

Klein (1984), compared the activity of a commercial formulation of

chlorpyrifos (Dursban) with that of 5 of its new slow-release

formulations (C-20A, C-20B, 48H, GGA1 and GGA2) against larvae of

Spodoptera littoralis in the laboratory and against eggs and small larvae

in cotton fields. He found that in the laboratory the commercial

formulation was effective for a maximum of 1 day against young larvae

only (1st- and 2nd-instars). All the slow-release formulations except 48h

gave good results against both small and large larvae, for periods between

3-5 days (for 5th-

and 6th-instar larvae) and 7-9 days (for 2nd-instar larvae).

However, in the field, the slow-release compounds were not superior to

the commercial formulation.

Kassem et al.(1985), found that all tested insecticides gave initial

kill to cotton leafworm of 95-100%, except AC222705, Bactospiene,

Bolstar+GM(0.7 1+1.2 1/f, tank mix), Dowco, KZ143, Padan, PH 66-13,

SIR 455 and Tamaron GM. The descending order of the insecticides

which had high residual activity against cotton leafworm between 70% -

90% mortality after 12 days was CGA74055, AC7802, Sumicidin ( 0.75

5


1/f ) CCN52( 0.6 1/f ), SH 488, Hostathion, Bolstar + GM ( 0.91 + 0.8 1/f

,tank mix ) and Bolstar 720.

Ghattas and EL-Keie (1986), investigated the extent of resistance

to certain organophosphorus insecticides namely sulprofos, chlopyrifos,

phosfolan, tiazophos, and profenofos on a field population of the cotton

leaf-worm collected from Minufiya Governorate, Egypt. They indicated

that the initial kill of all tested insecticides was not less than 90%

mortality during the five control seasons. They observed that the extent of

resistance varied considerably among the different cases investigated, the

population was obviously of high level of resistance by time to sulprofos

and chlopyrifos, moderate to profenofos and triazophos and less

pronounced to phosfolan and mephosfolan.

Zidan et al. (1987),studied the ovicidal efficiency of certain

organic insecticides and three mineral oils against the egg stage of

Spodoptera littoralis under laboratory and field conditions. They proved

that the evaluated organic insecticides varied tremendously in their

ovicidal potencies and cypermethrin seemed to be the most effective

materials, but thiodicarb was the least one in this respect. The mineral oil

KZ 4 showed the highest efficiency among the tested oil preparations.

Most of the tested materials gave satisfactory ovicidal effect under

laboratory conditions, while the efficiency was drastically decreased

under field conditions. Most of the evaluated insecticides-mineral oil

mixtures potentiate the activity in varying degrees. KZ 2 was the most

strong potenting agent among the tested organic insecticides in relation to

their ovicidal activities against S. littoralis. They also showed that adding

mineral oil to each of the most evaluated organic insecticides,

significantly increased the ovicidal effect under field conditions.

6


El-Ghareeb and Mannaa (1989), studied the insecticidal potency

and development of resistance in the cotton leafworm in upper Egypt.

They reported that the synthetic pyrethroids, deltamethrin, cypermethrin,

fenvalerate and flucythrinate had respectively the most toxic effect on the

two strains (Sohag and El-Minia) during 1986, 1987, and 1988 seasons.

Among the anticholinestrase insecticides, the oxime-carbamate,

methomyl was more toxic than the organophosphorus insecticides

(chlorpyrifos, phosfolan, profenofos, and sulprofos). In general, the two

strains exhibited higher level of resistance to the tested organophosphates

and methomyl. This was correlated with a slow rate of developing

resistance to the synthetic pyrethroids.

Rizk et al. (1990), investigated the effects of various insecticides

on eggs and larvae of Spodoptera littoralis. Treated cotton leaves were

fed to larvae in the laboratory and egg masses in the field were sprayed

with the insecticides. Hexaflumuron and diflubenzuron were ineffective

as ovicides. Chlorpyrifos (480 g a.i./fed.) gave complete controlthe pest,

irrespective of the age of the eggs to which it was applied. Residues of

hexaflumuron and diflubenzuron persisted longer than residues of

chlorpyrifos and had the greatest residual toxicity to larvae.

El-Dahan. (1991), studied the resistance status of Spodoptera

littoralis Boisd. to insecticides in Kafr El-Sheikh Governorate during

1984-1988 cotton growing seasons. He found that Spodoptera littoralis

field strain developed high resistance to pyrethroids, organophosphates

exemplified by chlorpyrifos, methamidophos and profenofos which were

comparatively more effective . On the other hand, the oxime carbamates

thiodicarb and methomyl were less potent than organophosphate and

produced levels of resistance that exceeded the allowed critical level (10-

folds).

7


Marenco et al.(1991), evaluated the residual activity of methomyl

and thiodicarb against fall armyworm; Spodoptera frugiperda larvae at

various post-application intervals in laboratory. Bioassay was done using

leaves of field-treated sweet corn . They reported that mortality of second

and fifth instar larvae were not significantly different from either of the

two insecticides. Foliage fed to larvae 3 hrs. after application (day 0)

resulted in 50-60% mortality for methomyl and >95% mortality for

thiodicarb. Mortality resulting from methomyl residues ranged from 5-

50% and 1-15% at 1 and 2 days post-application, respectively. Larvae

exposed to thiodicarb-treated foliage suffered >90% mortality for the first

4-5 day and 70-80% on day 9. thiodicarb residues induced about 50%

mortality of fifth instar larvae on day 14.

Mourad et al .(1991), studied the efficacy of eighteen insecticides

against the second and fourth instar larvae of the cotton leafworm

Spodoptera littoralis Bosid. in the field using two sprays. They found that

tested pyrethroids were the most toxic based on residual activity,

followed by organophosphorus and carbamate insecticides, Karate was

the most effective pyrethroid while Cybolt was the least in residual

activity. Hostathion was the most effective organophosphorus insecticide

but Dursban was the least, Cyolane, Cytrolane and Curacron were

moderately effective.

The efficacy of pesticides against cotton pests was evaluated by

Shalaby et al.(1991), according to the official methods of the Egyptian

Ministry of Agriculture and Soil Reclamation. They found that Omega

10% (cypermethrin) at 0.3 L/fed. gave the highest mean residual effect for

3 weeks, beside an excellent initial kill, The IGR- traditional insecticides

mixtures showed highly promising effects against 2nd and 4th instar larvae.

Mon 516, furathiocarb + IGR and methidothion + IGR gave excellent

8


initial and moderate residual effects, being higher than 90%. Field

evaluation indicated clearly the superiority of the IGR-profenofos

treatment.

Van Laecke and Degheele. (1991), studied the synergism of

diflubenzuron and teflubenzuron in larvae of Beet armyworm Spodoptera

exigua (Lepidoptera : Noctuidae). The results obtained showed that LC50

for fourth and fifth instars treated with diflubenzuron were 295 and 16

mg/liter, respectively ; for teflubenzuron, LC50

` s were 42 and 7 mg/liter,

respectively . The synergists profenofos and s,s,s-tributylphosphoro-

trithioate (DEF) enhanced the toxicity of diflubenzuron in fourth instars

nine and six folds , respectively. diethylmaleate decreased the LC50

five

folds, whereas piperonyl butoxide was responsible for only a three-folds

reduction

Abou-Kahla et al. (1992), studied the impact of several sequences

of insecticides against certain cotton pests and the associated predators in

cotton fields. They found that the sequence which includes Tamaron

combi, Cyanox and Larvin liquid gave the highest reduction percent

against the cotton leafworm (81.93%).The insecticides sequence

programs (Decis, Cyanox, Larvin liquid and Tamaron combi) and (Larvin

liquid, Tamaron combi, Decis and Cyanox) were the leading programs, as

they induced more efficient pest control, low efficacy on predator insects

and gave high cotton yield.

El-Ghareeb. (1992), studied the synergism of five insecticides

representing the pyrethroids, deltamethrin and flucythrinate and

anticholinesterase insecticides, methomyl, phosfolan and profenofos in

both larvae of Spodoptera littoralis (Boisd.) and adult of Coccinella

undecimpunctata L. by two inhibitors of detoxifying enzyme, the oxidase

9


inhibitor piperonyl butoxide (pb) and the esterase inhibitor TPP (triphenyl

phosphate). He indicated that, for the pyrethroid insecticide, deltamethrin,

it was the most toxic insecticide tested against the two species, although

there was no considerable difference in synergistic ratio between the two

pyrethroids on the same species, by using the same synergist. For the

anticholinesterase insecticides, on S. littoralis, methomyl was about 16.0

and 14.0 fold more potent as AChE inhibitor than phosfolan and

profenofos, respectively. Although methomyl was more susceptible for

detoxifying enzymes than phosfolan and profenofos in S. littoralis.

Methomyl exhibited higher toxicity than the two OP insecticides tested to

C. undecimpunctata.

El-Ghareeb and Mannaa.(1992), applied pyrethroids (trans-

permethrin, cis-permethrin, deltamethrin, cypermethrin, fenvalerate and

flucythrinate), an organophosphate (chlorpyrifos) and a carbamate

(methomyl) in the laboratory to 4th-instar larvae of Spodoptera littoralis.

Treated larvae were held at 22, 30 or 38°C and mortalities were recorded

after 24, 48 and 72h. They found that the toxicity of all the pyrethroids

except flucythrinate increased with decreasing temperature. It is

concluded that cyclopropane carboxylic acid esters have a larger negative

temperature coefficient than the methylbutyric acid esters. Chlorpyrifos

and methomyl showed positive temperature-toxicity relationships.

Mannaa and El-Ghareeb. (1992), studied the comparative

efficacy of variable and constant temperatures on the toxicity of certain

insecticides against cotton leafworm. They evidenced that the insecticides

tested, under the constant post-treatment temperatures (22°C, 30°C , 38°C)

and under the four post-treatment temperatures cycles, [each cycle

included four temperature exposure periods , each period took 6 hr's.

cycle I (22°C, 30°C, 38°C, and 30°C), cycle II (30°C, 38°C, 30°C, and

10


22°C), cycle III (38°C, 30°C, 22°C, 30°C), cycle IV ( 30°C, 22°C, 30°C,

38°C),] could be arranged in descending manner of toxicity as follows:

deltamethrin, cypermethrin, cis-permethrin, trans-permethrin, fenvalerate,

flucythrinate, chlorpyrifos and methomyl.

Flucythrinate (cybolt), which had been shown to be strongly

repellent to Spodoptera littoralis (Boisd.) and highly effective against

Pectinophora gossypiella (Saund.) and Earias insulana (Boisd.), gave

excellent control at application rates of 54-107 g a.i./ha against Aphis

gossypii Glov., Jacobiasca lybica (de Berg.) (Empoasca lybica),

Tetranychus cinnabarinus (Boisd.) and Thrips tabaci Lind. in several

vegetable crops (Abdel-Fattah et al. 1993).

Van Laecke and Degheele .(1993), carried out an extended

laboratory test with second instar larvae of Spodoptera exigua on Vicia

faba plants to determine the influence of synergists on the biological

activity of three benzoyl phenyl urea (BPUs).They found that the co-

application of piperonyl, an oxidase inhibitor, did not increase the activity

of diflubenzuron, teflubenzuron (at a concentration which resulted in 43%

survival) with diethyl maleate or dimethoate gave only 6.2 and 8.9%

surviving larvae, respectively. In addition, develotfourth instar larvae was

inhibited. The more stable teflubenzuron was synergized by both

compounds to a much lesser extent than diflubenzuron .None of the

synergists had a significant effect on the activity of hexaflumuron.

Hossain et al. (1994), tested binary mixtures of abamectin with

methidation, prothiofos, sulprofos, quinalphos, pirimiphos-ethyl and

chloropyrifos-methyl, respectively in ratio of 9:1, 4:1, 1.5:1 and 1:1

insecticide : abamectin for synergistic activity against the 4th

inster larvae

of cotton leafworm, Spodoptera littoralis (Bosid.). They found that

11


synergistic action was exhibited by all tested mixtures except the ratio 4:1

The dosage response data recorded after longer exposure period, 72hr.

post treatment revealed improvement in binary mixtures performance .

Binary mixtures of abamectin with either of prothiofos or/and

chloropyrifos-methyl exhibited remarkable antagonism at 24 and 72hr.

post treatment.

Rizk et al. (1994), studied the variation in susceptibility to

pyrethroids of Spodoptera littoralis (Boisd.) fed on different host plants.

They indicated that larvae reared on bindweed proved to be more

susceptible than those reared on all other hosts. Among the tested

pyrethroides, deltamethrin was the most potent insectcide, exhibiting

LD50

values ranging from 0.0960 to 0.0261 ug/larva. Based on the

relative susceptibility of larvae to pyrethroides, the hosts could be

arranged descendingly : bindweed > cabbage > clover > cotton.

1.2. Aphids

El-Sebae and Saleh. (1970), studied the aphicidial properties of

safer insecticides against Aphis craccivora on cowpea crop . They found

that Sevin, Malathion, Thiocron, and Dimethoate were efficient in

keeping the plants free of infestation for 10 days after spraying.

Mesbah et al. (1981), studied the toxicity of surfactant and

cypermethin surfactant combinations to the cabbage aphid, Brevicoryne

brassicae (L.). They reported that the activity of surfactants alone varied

and resulted in LC50

values ranged between 38.3-22000 ppm and 16.8-

1687 ppm at 24 and 48 h. , post treatment, respectively. The most active

compound was egyptal followed by nestapon, spane 20 and hamadol

whereas especrin and tween 20 exhibited the least aphicidal activity .

12


Tests with surfactant/insecticide combinations indicated higher

synergistic action in their aphicidal activity.

Neubauer et al .(1982), studied the foliar residues and toxicity of

three systemic insecticides to Aphis citricola, applied to soil in a citrus

grove . They found that the effectiveness of the treatment was affected

mainly by the rate of accumulation of toxicant in the leaves. At 18 g a.i.

per tree, the greatest residues found in the leaves were 106, 12.2 and 1.3

ug/g fresh weight for aldicarb, ethiofencarb and dimethoate, respectively,

ethiofencarb was the least toxic to the aphids.

Zein et al. (1982), tested nine insecticides related to different

chemical groups against Aphis gossypii (Glov.). They found that insect

location affected its susceptibility to the tested compounds. Also the

susceptible strain of the insect reared on different hosts, varied in its

sensitivity towards the tested insecticides. The temperature also affected

the susceptibility of the insect to the nine tested insecticides.

Jacob and Nair. (1983), showed that cypermethrin at 0.01 and

0.02%, metasystox [demeton-S-methyl] and monocrotophos both at 0.03

and 0.05% were highly effective and persistent against Aphis craccivora

infesting hollyhock. Lindene, phenthoate and phosalone were less

effective.

Kady, et al. (1983) evaluated six formulated insecticides under

laboratory conditions against Aphis gossypii. cypermethrin proved to be

the most toxic compound followed by triazophos , phosalone, methomyl

(Lannate), vamidothion and methomyl (Nudrin) in a descending order.

Two weeks after field application, cypermethrin, triazophos and

phosalone were maintaining excellent aphid moralities although

13


methomyl (Nudrin) and vamidothion were apparently losing

effectiveness.

Shanab et al. (1983), evaluated the joint effect of six herbicides

and five insecticides on the population density of Aphis gossypii and

Thrips tabaci under field conditions. They found that the application of

herbicides alone slightly decreased the population density of both insect

while insecticide treatment greatly suppressed the infestation and all

insecticides seemed to be highly effective. In addition, it was obvious that

the best combined effect on aphids population could be obtained in the

case of pendimethalin as herbicide and cypermethrin, phosalone and

triazophos as insecticides..

Singh and Sircar. (1983), studied the toxicity of insecticides

against 8 species of aphids . The most toxic compounds against Aphis

craccivora, A. gossypii, Brevicoryne brassicae, Dactynotus carthami

[Uroleucon carthami], Lip Aphis erysimi, Myzus persicae and

Rhopalosiphum maidis were phorate, dimethoate, phosphamidon,

dimethoate and carbaryl, respectively. Some evidence of resistance to

insecticides was found, and susceptibility varied with the food-plant.

Sagar and Jindla. (1984), tested various compounds as 0.005%

sprays applied from a knapsack sprayer. The quickest results were

obtained with monocrotophos, which killed 98% of the aphids within 24

h. of treatment, but chlorpyrifos, dimethoate and demeton-s-methyl

(Metasystox) also eliminated the aphid populations within a week.

Malathion and dichlorvos gave less satisfactory results.

Thakkur et al. (1984),studied the chemical control of aphid, Aphis

craccivora Koch. They found that all the insecticides were effective

against the pest as compared with untreated plots. The most effective of

14


the compounds 72 h. after treatment were dimethoate and phosphamidon,

while demeton-methyl and fenvalerate had an early knockdown effect.

Dimethoate remained the most effective 7 days after treatment.

The chemical control of spotted alfalfa aphid Therio Aphis trifolii

(Monell) on lucerne with reference to conservation of coccinellid

predators was studied by Sandhu. (1986), He found that five days after

spraying, oxydemeton-methyl at 75 and 150 g a.i./ha. and dimethoate and

phosphamidon at 150 g a.i./ha. were effective. Malathion was relatively

ineffective. The insecticides did not appear to be very toxic to adults of

the predatory coccinellids, Menochilus sexmaculatus, Brumus suturalis

[Brumoides suturalis] and Coccinella septempunctata.

Zein et al.(1987), carried out laboratory and field experiments to

evaluate the toxicity of seven pesticides against red spider mites,

Tetranychus cinnabrinus and aphids, Aphis gossypii. Results of the

laboratory and field studies showed that fenitrothion proved to be more

potent to aphids.

Kansouh et al. (1988), found that the application of oleodiazinon

(10% diazinon + 90 % mineral oil ) at rates of 5,10 and 20 ml /gal. water

was highly effective for controlling aphids and led to the reduction of

mosaic disease on squash plants.

Kassem et al.(1988), carried out a field evaluation of thirteen

insecticides treatments against the cotton thrips, Thrips tabaci (Lind.)and

the cotton aphid , Aphis gossypii (Glover) on the cotton seedlings . They

classified the efficacy of these insecticides to four groups. The first group

had very high effect (90 % reduction),the second group had high effect

(80% reduction ),the third group had moderate effect (70% reduction),

and the fourth group had low effect (less than 70% reduction).Omethoate

15


gave very highly effect against the cotton aphid, also vamidothion and

thiometon caused high effect while the other insecticides gave moderate

and low effectiveness.

Zidan et al. (1988a), studied the foliar fertilization affecting the

performance of certain aphicides and acaricides under laboratory and field

conditions. They found that the insecticides pirimicarb and malathion

were shown to be very effective against both nymphs and adults of A.

gossypii. The foliar fertilizers greatly reduced the aphicidal activity of

pirimicarb and malathion.

Hussein and Fong. (1989), found that the LC50

s (in mg/ml) of

dimethoate and (in brackets) malathion were 14.8 (17.8) for Menochilus

sexmacu [Cheilomenes sexmaculata], 75.9 (95.5) for Aphis craccivora

and 206.3 (239.8) for A. gossypii. Both insecticides reduced the fecundity

and adult life span of the predator. In the presence of dimethoate, the

level of prey population was influenced by the predator-prey ratio and the

dosage of insecticide used.

Zidan et al.(1989a),studied the relative efficiency of certain

insecticides on different developmental stages of the oat cherry bird

aphid, Rhopalosiphum padi (L.). They found that the synthetic pyrethroid

fenvalerate seemed to be the most superior compound against the

different nymphs and adult stages of oat cherry brid aphid R. padi (L.),

followed by the OP, monocrotophos and dimethoate. Methamidophos was

the inferior insecticide in this respect.

Zidan et al.(1989b), studied the effect of sublethal concentrations

of certain insecticides on the biotic potential of S. graminum during

successive generations. They obtained that, the tested insecticides had no

considerable effects on the nymph duration of S. graminum however , the

16


adult longevity was generally decreased at different rates according to the

chemical nature of tested compounds. Pirimicarb seemed to be the most

effective insecticide followed by pirimiphos-methyl, but fenvalerate was

the least one in this respect. They proved that also fenvalerate was the

superior insecticide in reducing the reproductive capacity of S.

graminum aphid , both of pirimiphos-methyl and pirimicarb had no

significant influence in this respect.

Ascher and Nemmy. (1990), determined the ovicidal activity of 61

organophosphorus (OPs), 10 carbamate and 13 other insecticides in a

primary screen against 0- to 1-day-old eggs of the Spodoptera littoralis

using an egg-dip bioassay (at 100 mg a.i./litter). Eleven of the OPs

(including Bolstar [sulprofos], Dursban [chlorpyrifos] and Guthion

[azinphos-methyl]) and 2 carbamates (CGA 45156 [thiodicarb] and

Larvin [thiodicarb]) gave 100% mortality. Insecticides which gave >90%

mortality were Baythion [phoxim] and Curacron [profenofos]. Most of the

compounds which were active in the primary screen were retested at

lower concentration ; the best results were obtained with Dursban.

Darwish et al. (1990), studied the efficiency of five chemical

compounds against the duranta aphid, Aphis punicae (Pass.) (Homoptera,

Aphididae) and its predator Cydonaia vicina var. nilotica (Muls.)

(Coleoptera. Coccinellidae). They found that profenofos (Curacron) and

fenvalerate (Sumicidin) were the most pronounced toxicants, XRD 473

was moderate while thuringienisn and phenoxycarb were the least

effective ones. There was a positive relationship between the toxicity of

pesticides used and the effect on the reproductive and survival potentiality

of Aphis punicae (Pass.).

17


Hogmire et al.(1990), reported that Aphis pomi was more

susceptible to esfenvalerate and methomyl but less susceptible to

azinphos-methyl than was A. spiraecola. The susceptibility to endosulfan

and chlorpyrifos was not significantly different between the 2 species. For

A. pomi, order of insecticide toxicity was esfenvalerate > methomyl >

endosulfan > chlorpyrifos > azinphos-methyl. For A. spiraecola, order of

toxicity was esfenvalerate > methomyl > azinphos-methyl > chlorpyrifos

> endosulfan.

Mcpherson and Bass (1990), investigated the control of red and

green morphs of tobacco aphids Myzus nicotianae (Homoptera:

Aphidididae) in flue-cured tobacco in field and labortory. In field

conditions acephate provided a good control of both morphs at 0.75 and

0.5 lb/acre, and fair to poor control of both morphs at 0.25 and 0.125

lb/acre. Methomyl, endosulfan, and endocide plus (endosulfan plus

parathion) also provided effective control of both morphs. Malathion,

oxamyl, and tralomethrin provided significantly better control of the

green morph than the red morph. Micro-encapsulated methyl parathion

was totally ineffective in controlling either green or red color forms . In

laboratory tests, the residual activity of acephate provided high mortality

and a reduction in live nymphs produced for up to 14 days after

application for both morphs, although these effects were more

pronounced with green aphids.

Rajput et al. (1990), studied the efficacy of systemic insecticides

against aphid on mustard crop. They found that emulsifiable concentrate,

Dimecron and Metasystox were significantly more effective when

compared with Anthio and Pirimor. Temik was comparatively more toxic

and persistent followed by Thimet and Solvirex in the granular

formulation.

18


Zaman. (1990), studied the evaluation of soil-applied granular

systemic insecticides against the cotton aphid on okra in Swat. Aldicarb

G (2.0 Kg/ha), mephosfolan G (0.9) and thiofanox G (1.25) were

reapplied as side-dressing just at the middle level of ridges after 73 days

of sowing of okra (cv. T-13). All the insecticides significantly reduced the

cotton aphid, Aphis gossypii, population during the season. Aldicarb was

superior to the other insecticides in its effect against the aphids.

Zeitoun et al. (1990), determined the effectiveness of 5 insecticides

i.e. malathion, dimethoate, profenofos, primiphos- methyl, omethoate and

mixture of pirimiphos-methyl and dimethoate to control sucking pests of

potatoes. They showed that profenofos at rate of 750 cm/F. proved to be

the potent on the sucking pests Aphis gossypii, Bemisia tabaci and

Empoasca lypica with an average general reduction of 85.06%. The other

compound gave reductions ranged from 69.7 to 82.5%.

Khattak and Hammed.(1991), studied some chemical control trial

against rape aphid, Brevicoyne brassicae L. They found that the

effectiveness of insecticides against rape aphid (B. brassicae) followed

the order: Atabron > Azodrin >Decis-D > Monitor > Trebon . highest

yield of Brassica napus was obtained with Atabron.

Mcleod. (1991) , indicated that at 24°C , application of five

systemic insecticides to the top surface of spinach leaves resulted in rapid

mortality of green peach aphid, Myzus persicae (Sulzer), on the ventral

side of treated leaves. Mortalities were generally delayed and reduced at

lower temperatures of 16 and 8° C. Systemic activity of dimethoate,

pirimicarb, phosphamidon, and acephate was low. Dimethoate failed to

provide statisfactory aphid supperssion at any tested temperature.

19


RH7988 appeared to be translocated rapidly at each tested temperature

and was effective in suppressing aphid on non treated leaves.

Rizk and Kamel. (1991), evaluated the effectiveness of Reldan

bioencapsulated formulation, Reldan EC, Dursban, and Dennet in cotton

field . They found that Dennet and Reldan bioencapsulated at rate of 400

g.a.i./fed. gave an immediate effect on aphids after 3-days of application

in comparison with Reldan EC (reduction percent values of 100, 99.8,

and 67.6 were recorded respectively).

Shalaby et al. (1991), evaluated the efficacy of pesticides against

cotton pests according to the official methods of the Egyptian Ministry of

Agriculture and Soil Reclamation . They found that Marshal 25 % at 300

g/fed and 900g/fed controlled excellently aphids and thrips (in early

season). However Polo 50% at 200 ml/fed. gave excellent control to

aphids and acceptable control to whitefly in the late season, but failed to

reach an acceptable efficacy level against aphids in the early season.

Sharma et al. (1991), determined the effect of the insecticides,

dimethoate (applied as Rogor at 0.40 kg a.i/ha), endosulfan (applied as

Thiodan at 0.70 kg a.i./ha), monocrotophos (applied as Nuvacron at 0.40

kg a.i./ha), fenvalerate (applied as Sumicidin at 0.20 kg a.i./ha) and

cypermethrin (applied as Ripcord at 0.04 kg a.i./ha) on Aphis craccivora

infesting Lathyrus sativus, lentils and chickpeas, and on its associated

coccinellid predator complex. Cypermethrin was the most effective

insecticide against A. craccivora on all 3 crops, whereas dimethoate was

the most toxic insecticide to coccinellid larvae on all 3 crops. Dimethoate

applied to L. sativus, dimethoate and monocrotophos applied to lentil, and

monocrotophos applied to chickpeas were all highly toxiccoccinellid

adults. Fenvalerate, followed by endosulfan applied to L. sativus,

20


cypermethrin and endosulfan (applied to lentils), and endosulfan (applied

to chickpeas) were considered to be the most selective insecticides for

aphid control.

Srivastava et al. (1991), evaluated six insecticides viz ,

phosphamidon, methamidophos, monocrotophos, pirimacarb, formothion,

and phenthoate (each at 0.025% concentration) for their relative efficacy

against mustard aphid Lip Aphis erysimi in field condition . They reported

that monocrotophos gave maximum yield followed by phosphamidon.

Maximum cost benefit ratio was recorded in phosphamidon 1:55:39 for

the first year and 1:45:52 for the second year. Therefore,

Phosphamidonmay be recommended as one of the cheapest insecticides

to combat mustard aphid L. erysimi.

Abdel-Wahab and Mohamed. (1992), evaluated the toxicity of

malathion, pirimiphos-methly, profenofos, carbosulfan and pirimicarb

against field and laboratory strains of the greenbug, Schizaphi graminum

(Rondani). They found that profenofos and malathion were the most

potent compounds against the field population, whereas pirimicarb was

the least toxic one.



cotton fields. They found that the sequence which contain

organophosphorus compounds had the higher effects against Empoasca

lybica and Aphis gossypii. while Bemisia tabaci was more affected by

treatments of Tamaron combi, Decis and Cyanox. The insecticides

sequence programs (Decis, Cyanox, Larvin liquid and Tamaron combi)

and (Larvin liquid, Tamaron combi, Decis and Cyanox) were the leading

21


programs, as they induced more efficient pest control, low efficacy on

predator insects and gave high cotton yield.

Bulut and Kedici. (1992), tested oxydemeton-methyl (Metasystox

25 EC), parathion-methyl (Folidol M-35) and pirimicarb (Pirimor 50 WP)

against Aphis pomi on apples in Turkey during 1988-90. Oxydemeton-

methyl, parathion-methyl and pirimicarb were effective.

Dimetry and Marei. (1992), studied the susceptibility of adults of

the cabbage aphid, Brevicoryne brassicae and two of the most important

predators, Coccinella undecimpunctata and Chrysopa carnea to

pesticides . Most of the organophosphorus and carbamate compounds

tested were toxic to the aphids. Profenophos was highly toxic to the adult

virginoparous aphid at LC50

level. Malathion demonstrated the highest

toxicity to the aphids. The application of profenophos for the cabbage

aphid at LC50

level exhibited no toxicity for Coccinella but was highly

toxic for Chrysopa. Pirimicarb proved to be the most selective of the

compounds tested for the control of aphids when it was used at a

concentration that caused 50 per cent mortality as it was found to be

unharmful to both Chrysopa and Coccinella. Dimethoate, on the other

hand, was found to be very harmful and highly toxic to both Coccinella

and Chrysopa adults and would probably be destructive to those natural

enemies in the field. Laboratory evaluation indicated that both pirimicarb

and malathion have potentials for use in IPM program for vegetable pest

control.

El-Defrawi and Abd El-Azim. (1992), studied the chemical

control of cowpea aphid ,Aphis craccivora Koch on Faba bean crop .

They found that , Pirimor (pirimicarp) with the two recommended rates,

Sumithion and Ekation displayed similar adequate bioactivity and were

22


significantly superior to all the tested compounds. The two insecticides ,

Actellic and Oki revealed lower toxicity, Malathion was the least toxic

compound and exhibited poor aphicidal action.

El-Sayed and El- Ghar. (1992), tested five insecticides at standard

and reduced rates against Bemisia tabaci and Aphis gossypii on

cucumbers, and the non target effects on some natural enemies were

assessed. They reported that The aphelinid parasitoids Eretmocerus

mundus and Prospaltella lutea [Encarsia lutea] were the most important

primary parasitoids of pupae of B. tabaci, and percentage parasitism was

slightly affected by insecticide application. However, all insecticides and

dosages caused severe suppression of the emergence of adult parasitoids,

and adult life span was heavily reduced. Populations of the aphid were

extremely reduced, especially by ethiofencarb and diafenthiuron.

Populations of the predators Chrysopa carnea [Chrysoperla carnea],

Coccinella septempunctata and Syrphus spp. were reduced in all treated

plots. However, ethiofencarb at rates as low as 208.4 g a.i./100 liters

provided equally as effective aphid control and conserved predator

numbers. Prothiofos and chlorpyrifos-methyl as low as 166.7 g a.i./100

liters kept aphid numbers below those in untreated plots and caused a

smaller reduction in the combined population of insect predators.

Gubran et al. (1992), studied the resistante in cotton aphid Aphis

gossypii (Glov) in the Sudan Gezira. They reported that the aphids were

found to be resistant to eight insecticides tested. There was no

enhancement of carboxylesterase activity and thus it was not a cause of

resistance in these species. First-order inhibition Kinetics of AchE with

pirimicarb revealed that resistance towards primicarb in Sudanese strain

was caused by modified AchE which had a reduced affinity (higher K_

value) and poor carbamylation ability (lower K2 value) for pirimicarb.

23


Kerns and Gaylor.(1992), studied the behavior of cotton aphid

exposed to sub lethal doses of three insecticides. They found that cotton

aphids, Aphis gossypii, that were not exposed to insecticides spent a

greater percentage of time feeding than walking. Aphids, treated topically

or exposed to residues of LC10

doses of cypermethrin or dicrotophos,

spent equal percentages of time walking and feeding. Topical application

and residual exposure to LC10

concentration of sulprofos increased

aphid activity with more spent walking than feeding.

Konar and Rai. (1992),studied the persistence of some

insecticides against insect pests of okra. They found that permethrin and

dimethoate proved to be more effective and persistent than other

insecticides against jassid and aphid, respectively followed by

dimethoate, carbaryl and malathion against jassid. and permethrin,

carbaryl and malathion against aphid.

Nielsen and King. (1992), showed that Pageant DF (active

ingredient of chlorpyrifos) was as effective for controlling Aphis pomi,

Chionaspis heterophyllae, Fenusa pusilla, larvae of Pyrrhalta luteola and

Neodiprion sertifer, and lepidopteran larvae feeding externally on

ornamental plants as Dursban 4E (another formulation of chlorpyrifos).

Shalaby and Ramadan. (1992), studied the efficiency of

Sumicidin, Meothrin, Cymbush and Palythrin against Aphis gossypii

(Glov.) and Bemisia tabaci (Genn) on okra plants. They found that all the

tested compounds exhibited reasonable insecticidal action against the two

insect species especially against aphids as compared with the whitefly

which was more tolerant. Percentages of reduction in infestation ranged

between 68% - 100% and 55% 81% in case of aphids and whitefly,

24


respectively, after 11 days form the onset of insecticidal application after

which sharp decrease in activity took place. Meothrin revealed the highest

bioactivity against the two insect species.

Wiles and Jepson. (1992), studied the susceptibility of a cereal

aphid pest and its natural enemies to deltamethrin .They found that the

range of LD50

values varied between 0.8 and 232 ng/arthropod and 0.76

and 66.17 mg/g body weight for the species tested. The linyphiid spider

Erigone atra (Black wall) was the most intrinsically susceptibility

predator species. There were significant differences in susceptible

between species of different orders, i.e .the Araneae, Hemiptera and

Coleoptera. Differences within the Coleoptera were closely related to

body weight, with the exception of the carabid beetle Demetrias

atricapillus (L.),which seemed to be tolerant to deltamethrin . The aphid

did , however , demonstrate a broader tolerance distribution than most of

the predators.

Abo El-Ghar et al.(1993), determined the effectiveness of 14

selected insecticides representing four insecticide groups

(organophosphates, carbamates, pyrethroides and Insect growth regulator

IGRs ) and four insecticide / IGR mixtures on cowpea insect pests and

their effects on certain beneficial insects . They found that after 3 days of

treatment , with phenthoate , isoxathion , cyanophos, carbaryl and

cypermethrin used at recommended rates . All insecticide treatments

significantly reduced the Bean Aphid, Aphis craccivora Kock, numbers

below that of the control. The prothiofos, isoxathion, pirimicarb and

fenpropathrin treatments provided continuing control to aphids through 21

days after spraying.

25


El-Hamady (1993) studied the compatibility of insecticides

(Actellic, Reldan, Selecron and Malathion) and chitin synthesis inhibitors

CSI (IKI17899, XRD473, Dowco 439 and SH777) and their interaction in

cowpea aphid Aphis craccivora Koch. He found that apart from SH777

all insecticides/CSI mixtures produced synergistic action at the three

mixing ratios (i.e. 2:1, 1:1 and 1:2 insecticide: CSI) against the tested

insect. However, Malathion was synergized when combined with all the

tested CSIs. A striking result is that, Malathion was strongly synergized

when mixed with IKI7899 or XRD473 at ratio 2:1 (LC50

value was

lowered from 480 to 65 and 69 ppm, respectively). In all cases, the

highest level of synergism was achieved at the mixing ratio 2:1 Apart

from malathion, all the tested insecticide/SH777 mixtures exhibited either

additive or antagonistic effect.

El-Maghraby et al.(1993a), studied the effect of pesticidal

application during the early and late season on the abundance of certain

predators associated with cotton plants . They found that application of

kelthane-s and methomyl against mites and aphids slightly reduced the

population of Orius spp, Chrysoperla carnea, Coccinella

undecmpunctata and Scymnus spp, percent reduction ranged between

10.6-17.9%. Also, they found that Orius spp was the most susceptible

species tolerated the action of the used insecticides.

El-Zemaity et al.(1993), studied the toxic effect and efficiency of

synthetic pyrethroids Sumi-alpha (contain Aa isomer) and Sumicidin

(included 4 isomers; BB, Ba, AB and Aa) against Aphis faba. Also,

compared their efficiency by carbamate insecticide, Pirimor. They

indicated that Sumi-alpha was the most effective insecticide followed by

Sumicidin then pirimor in the bioassay experiments .

26


Hussein et al. (1993a), studied the selective toxicity of certain

common insecticides used in controlling aphids to the predator,

Coccinella undecimpunctata and its prey cereal aphid, Rhopalosiphum sp.

they reported that the contact toxicity based on LC50

values demonstrated

the selectivity of profenofos, omethoate and alphamethrin to C.

undecimpunctata with selectivity factor (S.F) of 2.75, 1.42, and 1.166,

respectively. Methomyl showed slight selectivity (S.F 1.048). The relative

susceptibility of the predator and the aphid varied considerably among the

tested insecticides. The adult of C. undecimpunctata was more

susceptible to chlorpyrifos, malathion, pirimiphos-methyl and pirimicarb.

So they suggested that profenofos, omethoate, and alphamethrin could be

used in development of integrated management.

Park et al. (1993), tested five combinations of insecticides with

acaricides as wettable powders for their properties and use against mites

and aphids on apple trees in Korea Republic. Propargite plus chlorpyrifos,

propargite plus acephate, azocyclotin plus acephate, chlorpyrifos plus

clofentezine and chlorpyrifos plus bifenthrin, at 20 and 10%, 25 and 15%,

10 and 15%, 15 and 15% and 15 and 1.2%, respectively., chlorpyrifos

plus bifenthrin was found to be effective against Panonychus ulmi,

Tetranychus urticae, Myzus malisuctus [Ovatus malisuctus] and Aphis

spiraecola on apple in 1990-92.

El-Ghareeb and Nasser. (1994), demonstrated the toxicity of

twelve insecticides to five species of aphids collected from the field

during 1993 season. These species were Aphis gossypii (Glover),

Schizophis graminum (Rondoni), Hyalopterus pruni (Geoffroy), Aphis

craccivora (Kock) and Aphis neril (Boywr de fonscolomb). They found

that the toxicity of insecticides tested was different form one species to

another with chlorpyrifos-methyl, profenofos and parathion as the most

27


toxic compounds and methamidophos as the least toxic ones . The rest of

insecticides carbofuran, furathiocab, methomyl, pirimcarb, chlorpyrifos,

pirimiphos-methyl and flucythrinate occupied a moderate position.

Hyalopterus pruni was the most tolerant species followed by A. gossypii,

S. graminum, A. craccivora and A. nerii an ascending order of

susceptibility.

Abo-Sholoa et al.(1995), evaluated the effect of the application

numbers of three insecticides ; thiodicarb , chlorpyrifos and fenvalerate at

late cotton season against mainly bollworms besides sucking insects;

cotton aphid , Aphis gossypii (Glov), whitefly, Bimesia tabaci (Genn) and

jassed, Emposca lypica (Deberg) in two successive cotton seasons of

1992 and 1993. Each insecticide was applied once ,twice ,triple at two

weeks interval. They showed that both thiodicarb and chlorpyrifos had a

weak to moderate toxic effect and this effect was significantly affected by

the spray numbers . Fenvalerate failed completely as an aphicide.

Ibrahim.(1995), studied the development of insecticide resistance

in field populations of the cotton aphid (Aphis gossypii) Glover. He found

that resistance ratios and slopes of the probit regressions were variable

among years within insecticides, and resistance was not always consistent

among insecticides within an insecticide class. he indicated also higher

carboxylesterase activity in field population aphids compared with

susceptible aphids.

1.3. Whitefly, Bemisia tabaci :

Yein. (1983), found that applicaof 0.75 kg endosulfan,

monocrotophos, chlorpyrifos, dimethoate or quinalphos/ha. to Vigna

mungo plants was most effective againboth Bemisia tabaci and Pagria

signata [Colposcelis signata], while 0.75 kg phosalone or carbaryl/ha was

28


effective against Colposcelis signata only. Endosulfan was the most

effective and gave the highest increases in seed yield.

Salama et al. (1984), tested methomyl, decamethrin [deltamethrin],

diflubenzuron, chlorpyrifos and dimethoate in the laboratory , in different

sequences against sucking pests of cotton (Bemisia tabaci, Empoasca

lybica [Jacobiasca lybica], cicadellids, Tetranychus cinnabarinus,

aleyrodids, Aphis gossypii and other aphids) and some of their predators

(Amblyseius spp., Scymnus spp., Coccinella undecimpunctata and

Paederus alfierii). They reported that although more than 1 application of

the same insecticide is not permitted in cotton fields in Egypt for practical

pest control, 3 applications of the same compound were made

experimentally in field tests to ensure that the insecticides would affect

the predators. All combinations of insecticides (especially the repeated

ones) markedly reduced at least some species of predators. In general, P.

alfierii and Scymnus spp. were the most susceptible non-target species,

and deltamethrin was the compound most toxic to the predators. Against

the pests, the most effective combination was that of dimethoate,

methomyl and deltamethrin in separate successive applications, which

caused only slight mortality of predators.

Abdallah, et al. (1985), studied the effect of certain pesticides

against some non-target sucking pests infesting cotton .They found that

treatment of DC 702 ( in three sprays ) was the most effective one against

Tetranychus urticae (Koch.), whitefly , Bemisia tabaci (Gennandius),

and the jassid, Emopasca lybica (De Berg). Treatment of Curacron (in

three sprays) was as effective as that of DC 702 against the jassid and

whitefly, but was the least effective one against the spider mite.

29


El-Serwiy et al.(1985), studied the comparative efficacy of

different insecticides against some stages of whitefly Bemisia tabaci

(Genn) on Cucurbits. They reported that Supracid was more effective than

Actillic or Malathion on adults or eggs and larvae after 7 and 4 days post-

treatment, respectively with significant differences between the means of

their population reductions %.Eggs reduction may be attributed to the

direct effect of the insecticide against adults and newly hatched larvae

from eggs . Supracid spraying with concentration 0.2% four times at

intervals of 15 days led to an increase of 8607.5 Kg/ha. in fall cucumber

yield.

Fahmy et al. (1985), studied the effect of certain pesticides against

some non-target sucking pests infesting cotton. They found that all tested

treatments showed significant reduction in numbers of the considered

pests : the spider mite, Tetranychus utricae (koch.), the whitefly, Bemisia

tabaci (gennandius) and the jassid, Emopasca lybica (De Berg).

Treatment with DC 702 (in the three sprays) was the most effective one

against the examined pests. Treatment of Curacron (in the three sprays )

was as effective as that of DC 702 against the jassid and the whitefly.

Zanaty and El-Hawary.(1988), proved that pyrethroids

diminished the population levels of the whitefly , leaf hopper , aphids and

red spider mite and their predators in cotton fields . The oxime

carbamate , thiodicarb , was moderately effective against the sucking pest

and less effective against their predators.

Zidan et al.(1989c), studied the role of different insecticides

regime on the population density of mature and immature stage of cotton

whitefly , Bemisia tabaci (Genn.) in nursery and permanent field, with

special references to phytotoxicity and virus infection. In this respect,

30


methamidophos and fenvalerate/dimethoate treatment proved superior

influences / performances among the tested compounds.

Zidan et al.(1989d), studied the bio-residual efficiency of

methomidophos, butacarboxim, fenvalerate / dimethoate and fenvalerate /

fenitrothion mixtures against the adults of cotton whitefly Bemisia tabaci

on caged tomato seedlings in laboratory. They proved that the mixture

fenvalerate/dimethoate superior knock down as well as residual activities,

followed by methamidophos showing the LC50

s of 39.81 and 134.9 ppm,

respectively. Fenvalerate/fenitrothion mixture and butacarboxim showed

lower efficacy revealing the LC50

s of 436.5 & 457.1 ppm, respectively.

Darwish and Farghal. (1990), evaluated nine insecticides against

the cotton whitefly, Bemisia tabaci and associated natural enemies in

cotton fields of Assiut. They found that Buldock, Cyanox, EMA 2784 and

Tamaron combi showed the highest toxic effect against the whitefly.

Birlane and Marshal were the least effective ones. As for natural enemies,

Marshal and Tamaron combi were the least toxic compounds whereas,

Ekalux and Cyanox were the highest toxic ones. According to the relative

toxicity and residual effect of the tested insecticides against the whitefly

and associated natural enemies, the pyrethroid insecticide, Buldock gave

the best results.

The effectiveness of Reldan bioencapsulated formulation, Reldan

EC, Dursban, and Dennet in cotton field was evaluated by Rizk and

Kamel. (1991), They indicated that the bioencapsulated formulation of

Reldan gave good control against larval stage of B. tabaci (48.1) and

better than the recommended rate of Dennet (2.4) four weeks postspray.

The residual activity of the bioencapsulated formulation was reduced

considerably against the pupal stage of the whitefly and was similar to

31


Dennet through the four weeks post spray (Average reduction values were

58.8 and 55.3%).

Sadhakar and Paul. (1991), studied the efficacy of conventional

insecticides for controlling of cotton whitefly (Bemisia tabaci) and gram-

pod borer (Heliocoverpa armigera ) on cotton (Gossypium species).They

found that triazophos 0.05% and amitraz 0.05% effectively controlled

whitefly and gram-pod borer resulting in 53% more yield of raw cotton

compared with the control.

El-Sayed and El-Ghar. (1992), tested five insecticides at standard

and reduced rates against Bemisia tabaci and Aphis gossypii on

cucumbers. They reported that eggs of the aleyrodid appeared to be less

susceptible than larvae and pupae, and experienced a maximum of 66%

reduction; larval and pupal populations were significantly reduced in all

treated plots. For example, 10 days after the application of ethiofencarb,

diafenthiuron and chlorpyrifos-methyl, larval populations were reduced to

67, 50 and 68%, respectively., and pupal populations by 68, 69 and 75%.

Abbassy et al. (1993) studied the efficiency of certain natural and

synthetic pesticides in controlling white flies and mites on cucumber

grown under plastic tunnels. They found that Actellic and Reldan, were

the most effective pesticides against white flies followed by the

carbamate insecticides.

Abdel-Fattah et al. (1993), showed that flucythrinate (Cybolt)

gave excellent control at application rates of 54-107 g a.i./ha of Bemisia

tabaci (Gennadius) on soyabeans.

Abo El-Ghar et al.(1993), studied the impact of several

insecticides and insect growth regulators against certain insect pests of

32


cowpea and the associated beneficial insects. They found that most

insecticides treatments were not effective against cotton whitefly, Bemisia

tabaci (Genn.).The best control of the whitefly immature was obtained

after 3 days of spraying in plots received thiodicarb (76%) and

fenpropathrin (60%) .All selected insecticides and rates used had very

low residual effect against B. tabaci immature by 3 days post treatment.

Hussein et al. (1993b), investigated the toxicity of three

pyrethroids i.e. cypermethrin, fenvalerate and alfhamethrin, three

organophosphours insecticides (sulprofos, profenofos, chlorpyrifos) and

one carbamate (carbosulfan) to the adult and larval stages of laboratory

and field populations of Bemisia tabaci (Genn). They found that synthetic

pyrethroids (cypermethrin and fenvalerate ) were most effective against

adult and larval stages of both strains (adult and larvastages) and

appeared more resistante to all tested insecticides except carbosulfan.

Salem (1993), tested six insecticides against immature stages of B.

tabaci. The best control results were achieved with Danitol alone or

Applaud mixed with Actellic followed by Polo and Applaud separate.

Farrag et al. (1994), evaluated the relative toxicity of three

insecticides, Marshal 25% WP, Actellic 50% EC and Trebon 30% EC in

single continuous and alternating applications against the adult of

whitefly Bemisia tabaci (Gennadius) on cabbage plants. They found that

the percentage of reduction (%R) were 41.02, 42.60, and 87.37 with

Marshal, Actellic and Trebon, respectively after 7 days of the first

application. B. tabaci was more susceptible to Trebon in the

2ndapplication than Marshal and Actellic. Also, response of B. tabaci to

Marshal in the 2nd application declined after Trebon in the 1st one. No

33


reduction was obtained after 5 days of continuous use of Actellic in the

2nd

application.

Zidan et al. (1994) studied the bio-residual activity of certain

insecticides against the population density of whitefly, Bemisia tabaci

infesting cucumber plants in plastic houses . They indicated that the

important role of insecticide type and post-treatment period in

determining the population density of whiteflies . They classified the

insecticides into three groups based on their initial and residual

efficiencies against whitefly as follows : Actellic and Prempt which

proved to be the highly effective compounds , revealed 95.10 and 93.13%

reduction percentage, respectively . The second group includes admiral

and polo which gave reduction percentage of 71.12 and 67.3%,

respectively . The third group represent natural oil showing 58.18

reduction percentage. Efficacy of natural oil and insect growth regulator

increased by the elapse of time due to their latent effect.

El-maghraby et al. (1997). compared evisect natural insecticide

with recommended insecticides, Selecron 72% EC, Osbac 50% EC,

Admire 20% SL, Actellic 50% EC and Peldan 40% EC against whitefly

Bemisia tabaci on tomato plants. They found that All insecticides gave

hight efficiency against all whitefly stage ( mean %reduction ranged

between, 87.41 : 91.40- 90.17 : 93.09 and 88.32: 95.85 for adult,

immature and egg stage, respectively), also there were no siginificant

differenceis between insecticides.

1.4. Spider mites

El-Sebae et al. (1978), studied the toxicity of certain pesticides and

their combination against the red mite species Tetranychus

34


cucurbitacearum (Sayed). They found that of the chlorinated hydrocarbon

acaricides tested, dicofol was the most effective followed by

chlorobenzilate while tetradifon had the least toxic effect. Tamaron was

the most effective of the organophosphorus material followed by

dimethoate and then zolone which gave the least toxic action. Fundal and

Galecron were two formulation for the same active ingredient, however,

Galecron was nearly 1.5 times more toxic than Fundal. Cyolane was the

most effective insecticide and leptophos was the least while carbaryl

showed no toxicity. Karathan was found to be the most active fungicide

followed by benomyl and then morestan which gave the least among this

group. The two fungicides diathane M-45 and bavistin were non-toxic

and having no acaricidal activity. The higher potentation of pesticides

pairs on T. cucurbitacearum was leptophos + monocrotophos followed by

cyolane + chlorobenzilate and then by monocrotophos + tetradifon in a

descending order.

Mostafa.(1982), determined the susceptibility of eggs and adult

females of a strain of T. urtica originated from Rafah north Sinai

Governorate to seven pesticides. They observed that synthetic pyrethroids

( cypermethrin, decamethrin , and cybolt ) were the most effective as

adulticides, the LC50

S for adult females were 0.22, 0.34 and 1.06 ppm.,

respectively .The organophosphorus compound, Birlane showed the

highest ovicidal effect, Kelthane was intermediate in its toxicity to both

tested stages. The other organophosphorus compounds (Sumithion and

Rogor) showed low toxicity to the both stages.

Eissa et al. (1985),studied the impact of six different pesticides

(triflumuron, IKI-7899, isoxathion, Bacillus thuringiensis, carbophos and

phenthoate/oil) upon major citrus trees phytophagous and predatory mites

in Shebin El-Kom , Minufiya Governorate. They revealed that Bacillus

35


thuringiensis at 0.084%, phenthoate/oil at 1%, IKI-7899 at 0.375% and

triflumuron at 0.375% provided a good control (74-85% reduction) of

phytophagous mite, Tetranychus cinnabarinus (Boisd.). However, less

reduction of 68-75% in population of Brevipalpus californicus was

manifested by phenthoate/oil at 1%, isoxation at 0.2% and B.

thuringiensis at 0.084%. In general, B. thuringiensis, triflumuron,

phenthoate/oil and isoxathion at the tested rates were less toxic to the

predatory mites than the phytophagous mites.

Zein et al.(1987), carried out laboratory and field experiments to

evaluate the toxicity of seven pesticides against red spider mites ,

Tetranychus cinnabrinus and Aphids, Aphis gossypii . Results of the

laboratory and field studies showed that dicofol was more effective than

other tested compounds against mites.

Zidan et al. (1988a), studied the foliar fertilization affecting the

performance of certain aphicides and acaricides under laboratory and field

conditions. They found that adults of T. urticae were more susceptible

than eggs to dicofol. Tetradifon had a high ovicidal activity. Foliar

fertilizers reduced the performance of the acaricides.

Shoukry et al.(1989), studied the effects of 15 compounds from

different chemical groups against the different developmental stages of

standard colony of spider mite Tetranychus urtica. They found that: (1)

Tetradifon was the most potent compound against all ages of eggs

followed by dicofol. (2) Triflumuron was the most effective one against

the chrysalis stages followed by chlorofluazurn and flufenoxuron,

respectively.(3) Dicofol had higher effect against the adult stage followed

by profenofos.

36


The efficiency and persistence of certain insecticides on soybean

and broad bean plants was studied by Abdel-All et al. (1990), They found

that combinations of either chlorpyrifos or methomyl with two insect

IGRs tested (diflubenzuron and dowco-439) provided longer residual

activity for plant protection than IGRs alone. The efficiency of

chlorpyrifos plus IGRs was higher at 7 and 14 days after application on

aphids. The effectiveness of tested mixtures was in general lower on T.

urticae, at 21 days following application.

Labonowska, B. H. (1990a), studied the effectiveness of new

acaricides and some synthetic pyrethroids in the control of the two-

spotted spider mite (Tetranychus urticae koch) on strawberries.

Propargite WP formulation, cyhexatin, azocyclotin, bromopropylate and

standard acaricide-tetradifon showed the highest and satisfactory

effectiveness in the control of mites on strawberries. Fenpropathrin,

flucythrinate, flubenzimine, clofentezine and fenbutatin oxide (1 Kg/ha)

showed satisfactory activity, but within shorter time than tetradifon.

Labonowska, B. H. (1990b), reported that cyhexatin, azocyclotin

and fenpropathrin appeared to be very efficient in the control of spider

mites , even with very numerous populations of mites . Propargite ,

bromopropylate and flubenzimine gave better or similar results as the

standard acaricides, tetradifon, but a single spraying of this acaricide on

the bushes with very numerous population of adult mite did not gave

satisfactory reduction of mite population.

Labonowska, G. S. and Jesiotr.(1990), evaluated twenty-six

acaricides for the control of T.urticae on roses . They found that the most

effective specific acaricides were dienochlor, flubenzimine and

bromopropylate; and the non specific acaricides metamidophos.

37


Labonowska, G. S. et al.(1990), evaluated twenty-six pesticides

for the control of spider mites on gerbera. Dienochlor, cyhexatin,

bromopropylate and propagite applied 2-4 times during 90-114 days

controlled the spider mites better than standard acaricides.

Richardson and Lembright. (1990), compared new formulation of

chlorpyrifos (lock-on) at 0.5 lb a.i./acre with Dorsban 4E (chlorpyrifos) at

1.0 lb a.i./acre and Pounce (permethrin) at 0.1 lb a.i./afor the control of

Lygus sp., whitefly; Bemisia tabaci, Bucculatrix thurberiella,

Tetranychus spp. and Heliothis spp. In 1986 Lock-on was compared with

Lorsban, Pydrin (fenvalerate) and Ammo (cypermethrin) for the control

of pink bollworm, Pectinophora gossypiella, Lygus sp. and Tetranychus

spp. Control of Lygus sp., P. gossypiella and Heliothis spp. with Lock-on

was similar to that with the pyrethroids, but Lock-on gave better control

of B. thurberiella.

Smolarz et al . (1990), studied the control of the fruit tree, red

spider mite Panonychus ulmi Koch with fenpropathrin and flucythrinate

in apple orchard. They reported that both fenpropathrin and flucythrimate

showed a strong miticidal activity, which was comparable to that of

organotin acaricides. Thus, both might be recommended for controlling

insect pests in the orchard with no danger of increased mite injury.

Nowakowski and Szufa. (1990), studied the effects of insecticides

on the fruit tree red spider mite population (Panonychus ulmi Kock ).

They found that mite population was much more numerous on

pyrethroids permethrin, flucthrinate, fenpropathrin - treated plots than on

the other insecticide-treated ones. The number of eggs was the greatest in

permethrin-treated plots.

38


Kadir and Knowles . (1991), studied the toxicity of the thiourea

diafenthiuron to diamondback moths (Lepidoptera: Yponomeutidae), two

spotted spider mites (Acari: Tetranychidae), and bulb mites (Acari:

Acaridae). They found that diafenthiuron possessed low toxicity to two

spotted spider mites Tetranychus urticae ( 41.7% mortality at 500 ppm )

and was inactive against bulb mites, Rhizoglyphus echinopus (0 %

mortality at 1000 ppm). On the other hand they found that 3-(2,6-

diisopropyl-4-phenoxyphenyl)-1-tert-butylcarbodiimide(DFCD),

(diafenthiuron photodegredation product) was highly toxic to two spotted

spider mites ( LC50 8.9 ppm ), and bulb mites ( LC50 106.6 ppm ). Bulb

mites converted diafenthiuron-C14 to DFCD and to 3-(2,6-diisopropyl-4-

phenoxyphenyl) -1-tert-butyl urea which was non toxic.

Lawson and Weires. (1991), studied the management of European

red mite (Acari: Tetranychidae) and several aphid species on apple with

petroleum oils and an insecticidal soap . They reported that Sunspary 6E

and Volck supreme oils caused the greatest mortality of all material

tested. Sunspray 6E+ caused less mortality than did the Sunspray 6E and

Volck supreme but greater mortality than safer insecticidal soap

concentration, which caused only slightly greater mortality than the

distillated water check against over wintering Panonychus ulmi eggs .

Field application of the same materials by airblast sprayer and high-

pressure handgun caused less mortality of over wintering eggs than in the

laboratory study. Summer application of these materials were tested for

their ability to suppress mite and aphid populations throughout the

growing season. Applications of all materials provided significant control

of populations of P. ulmi, rose apple aphid, Aphis Plantaginea, and the

apple aphid, Aphis pomi, and the spirea aphid, Aphis Spiraecola.

39


Abbassy et al. (1993) studied the efficiency of certain natural and

synthetic pesticides in controlling white flies and mites on cucumber

grown under plastic tunnels. They found that from all of the tested

synthetic insecticides, Reldan only exhibited a considerable potential

acaricidal effect. As a specific acaricide, Tedifol was more potent than

Kelthane against the tested mites.

Fouly et al. (1995), studied the effect of different pesticide

application methods on mites inhabiting apple trees. They found that

spraying was the most suitable method to control the mites infesting plant

leaves where Cidial gave the highest level of effectiveness followed by

Basudin, Atabron and Selecron.

1.5 Beneficial arthropods.

Salman and Abd-El-Raof. (1978), studied the effect of certain

pesticides used against cotton pests on three predaceous insects and

honey bee workers. They found that all insecticides tested were more

toxic to the predaceous insects and honeybees (from 11 to 935 times as

toxic) than to S. littoralis. Similarly, 4 of the acaricides showed greater

toxicity to honeybees than to T. urticae although Curacron [profenofos],

Ekatin [thiometon], Kelthane [dicofol] and SAN 1551 (5-dimethylamino-

1,2,3-trithiane hydrogenoxalate) were more toxic to T. urticae. All tested

acaricides except Carbicron [dicrotophos] were more toxic to

Euteranychus orientalis than to honeybees.

Adashkevich and Kiriyak. (1983), studied the effects of various

insecticides on 2 natural enemies of the peach aphid Myzus persicae

(Sulz.) infesting tobacco and transmitting virus diseases to the crop. The

natural enemies were the seven-spotted ladybird [Coccinella

septempunctata L.] and Diaeretiella sp., and the insecticides were

40


menazon (Sayfos), phosalone, parathion-methyl (Metaphos), pirimicarb

(Pirimor), dimethoate (Bi 58) and fenitrothion (Metathion). Menazon,

pirimicarb and phosalone were least toxic to adults of C. septempunctata,

killing 24, 29 and 33% of individuals, respectively, after 10 days. After

10 h, fenitrothion had killed 83% of beetles, and by the 10th day had

caused total mortality; parathion-methyl and dimethoate caused 23 and

62% and 16 and 88% mortality, respectively.

Broadley. (1983), studied the contact toxicity of insecticides to

adults of the coccinellids Coccinella repanda and Harmonia

octomaculata, The toxicity in ascending order was: endosulfan,

chlorpyrifos, profenofos, methidathion, methomyl, monocrotophos,

cypermethrin and deltamethrin. The ascending order of toxicity for the 6

insecticides tested on Harmonia octomaculata was permethrin, sulprofos,

methidathion, chlorpyrifos, cypermethrin and deltamethrin. H.

octomaculata was less susceptible to methidathion.

Singh and Sircar. (1983), studied the toxicity of insecticides

against 8 species of aphids and the predacious coccinellid, Coccinella

septempunctata. They found that endosulfan, lindane and aphidan [S-

[(ethylsulfinyl)methyl] O,O-bis(1-methylethyl) phosphorodithioate] were

effective against aphids and relatively safe against C. septempunctata.

Krishnamoorthy. (1985), studied the toxicity of 15 pesticides used

at the recommended field rates to eggs, larvae and adults of Chrysopa

scelestes [Brinckochrysa scelestes], Eggs were sprayed directly, while

larvae and adults were exposed to dried residues on filter paper for 6 h. In

general, eggs were unaffected by the pesticides, but newly-hatched larvae

were susceptible to organophosphate and carbamate compounds.

41


Quinalphos, chlorpyrifos, malathion and dichlorvos were highly toxic to

both larvae and adults.

Patel and Vyas. (1985), evaluated the comparative toxicity of 10

insecticides to eggs of the neuropteran Chrysopa scelestes Brinckochrysa

scelestes, a predator of the noctuids Heliothis and Spodoptera, in the

laboratory . The decreasing order of toxicity of the compounds was

0.15% carbaryl > 0.05% malathion > 0.05% phenthoate > 0.05%

quinalphos > 0.07% endosulfan > 0.0125% permethrin > 0.01%

fenvalerate > 0.05% phosalone > 0.04% monocrotophos > 0.01%

cypermethrin.

Pree and Hagley. (1985), studied the toxicity of pesticides to

Chrysopa oculata Say (Neuroptera: Chrysopidae). They found that most

organophosphorus insecticides except phosalone were toxic to both stages

(first instars and adults). The synthetic pyrethroids, with the exception of

fenvalerate and flucythrinate, were generally more toxic than

organophosphorus insecticides. Adults and first instars were equally

susceptible to most insecticides. The acaricides and fungicides were not

toxic at the level tested. Methomyl and carbaryl were toxic to both stages,

but pirimicarb was almost non toxic. DDT and endosulfan were toxic

only at high concentrations. In the field, azinphos-methyl and phosmet

were toxic to larvae for at least 14 days. Permethrin killed larvae caged

on foliage for 10 days, whereas most survived on phosalone-treated

foliage 1 day after app.

Sandhu. (1986), studied the chemical control of spotted alfalfa

aphid Aphis trifolii (Monell) on lucerne with reference to conservation of

coccinellid predators. He found five days after spraying, oxydemeton-

methyl at 75 and 150 g a.i./ha and dimethoate and phosphamidon at 150 g

42


a.i./ha were effective. Malathion was relatively ineffective. The

insecticides did not appear to be very toxic to adults of the predatory

coccinellids Menochilus sexmaculatus, Brumus suturalis [Brumoides

suturalis] and Coccinella septempunctata.

Upadhyay and Vyas. (1986), studied the comparative toxicity of

various systemic insecticides and Malathion to the predatory coccinellids

Menochilus sexmaculatus and Coccinella septempunctata associated with

sucking pests Aphis craccivora and Empoasca kerri on groundnut in field

trials. A spray of thiometon applied against these pests at 0.03 and 0.05%

was found to be the least harmful to the coccinellids, followed by

demeton-O-methyl. Malathion at 0.05% was highly toxic to both

coccinellids. The comparative toxicity to coccinellids in ascending order

was thiometon < demeton-O-methyl < phosphamidon < monocrotophos <

dimethoate < malathion.

Wiktelius. (1986), tested the insecticides Pirimor G [pirimicarb],

Metasystox R100 [oxydemeton-methyl], Sumicidin 10 FW [fenvalerate],

Cymbush dg [cypermethrin] and Karate [[1a(S),3alpha(Z)]-isomer of

cyhalothrin] in the laboratory against predators of cereal aphids,

especially Erythraeus spp., lycosid spiders, the staphylinids Tachyporus

spp., the carabids Bembidion lampros, Harpalus rufipes and Pterostichus

melanarius and the Coccinellid Coccinella septempunctata. Pirimicarb

was the least toxic. The effects of the other insecticides differed between

test insects.

Bull et al. (1987), compared the toxicity of several insecticides

applied topically to adults of Microplitis croceipes (Cresson) , a larval

parasite of Heliothis spp . LC50

ranged from 0.008 to 0.658 ugm per

43


insect ; order of toxicity was: methyl parathion > dieldrin > malathion >

dimethoate > cis-permethrin > DDT > fenvalerate .

Zheng and Wing. (1987), studied the toxicity of pirimicarb,

fenvalerate, dimethoate and g-BHC [Lindane] to the cereal aphid Sitobion

avenae, its predator Coccinella septempunctata and bracing parasitoid

Aphidius avenae [A. picipes], as well as the effectiveness of the

insecticides against the aphid in the field. The LC50

s for pirimicarb,

fenvalerate, dimethoate and lindane were 0.99, 3.03, 6.53 and 109.2

p.p.m., respectively. Pirimicarb showed the highest selective ratio (natural

enemies LC50

:aphid LC50

) of the 4 insecticides tested. Field trials

indicated that pirimicarb at 105 g a.i./ha gave the best control of the aphid

and was the least harmful to the natural enemies, while fenvalerate at 60 g

a.i./ha was also effective in controlling the aphid and did little damage to

the natural enemies. Dimethoate at 180 g a.i./ha was effective in

controlling the aphid but was harmful to the natural enemies.

Applications of lindane at 225 g a.i./ha were harmful to the natural

enemies and the compound was not recommended for control of the

aphid.

Fayad and Ibrahim. (1988), examined the effect of insecticides on

predator numbers in cotton fields. Coccinella undecimpunctata was

highly affected by the 3 insecticides tested, Rup 962 [deltamethrin], DC

702 [chlorpyrifos and diflubenzuron], and Curacron [profenofos]. The 1st

insecticide was the most critical in the disturbance of Scymnus and Orius

spp. No significant differences were noted between the insecticides or the

interval between treatments. Paederus alfierii and spiders appeared to

tolerate the effects of the insecticides and were encountered in moderate

44


numbers throughout the study. Chrysopa vulgaris [Chrysoperla carnea]

was highly susceptible to all tested insecticides.

Baspinar and Uygun. (1990), studied the effects of some

insecticides on larvae, pupae and adults of Cryptolaemus montrouzieri

and pupae and adults of Coccinella septempunctata in the laboratory.

Methidathion, carbosulfan, furathiocarb, pirimicarb, fluvalinate and

summer petroleum oil were applied by direct spraying, dipping and film

methods to adults and by direct spraying only to larvae and pupae.

Methidathion, carbosulfan and furathiocarb were toxic, fluvalinate was

slightly harmful, and pirimicarb and summer petroleum oil were harmless

to all stages of both species. Larvae and pupae of Coccinella

septempunctata were more resistant to the insecticides than the adults of

both species.

Darwish and Farghal. (1990), evaluated nine insecticides against

the cotton whitefly, Bemisia tabaci and associated natural enemies in

cotton fields of Assiut. They found that Buldock, Cyanox, EMA 2784 and

Tamaron combi showed the highest toxic effect against the whitefly.

Birlane and Marshal were the least effective ones. As for natural enemies,

Marshal and Tamaron combi were the least toxic compounds whereas,

Ekalux and Cyanox were the highest toxic ones. According to the relative

toxicity and residual effect of the tested insecticides against the whitefly

and associated natural enemies, the pyrethroid insecticide, Buldock gave

the best results.

The efficiency of five chemical compounds against the duranta

aphid, Aphis punicae (Pass.) (Homoptera, Aphididae) and its predator

Cydonaia vicina var. nilotica (Muls.) (Coleoptera. Coccinellidae) were

studied by Darwish et al. (1990). They found that the application of the

45


tested compounds on the third larval instar of lady-birds, Cydonaia vicina

var. nilotica (Muls.) ensured that Sumicidin and XRD473 could be used

with a lethal concentration on the pest at 0.1 and 5 ppm., respectively

without any effect on the predator.

Hao et al. (1990) tested the eggs, 4th -instar larvae, pupae and

adults of Coccinella septempunctata with pirimicarb, phosalone and

monocrotophos at various concentrations using the leaf-soaking method.

The results showed that the eggs, larvae, pupae and adults reacted

similarly to the 3 insecticides. The toxicity index of pirimicarb and

phosalone in comparison with that of monocrotophos was 0.0483 and

0.3187, respectively., to eggs at 120h., 0.01088 and 0.4056 to 4th -instar

larvae at 24h., 0.0540 and 0.5990 to pupae at 216h., and 0.1406 and

0.0623 to adults at 24h., respectively., suggesting that pirimicarb and

phosalone were highly selective to different developmental stages of the

coccinellid.

Shukla et al. (1990), studied the comparative toxicity of some

insecticides to Coccinella septempunctata Linn. (Coleoptera:

Coccinellidae). They found that oxydemeton-methyl 25 EC at a

concentration of 0.040% was the most toxic and endosulfan 35 EC at a

concentration of 0.070% was the least toxic to both larvae and adults of

the coccinellid. The 4 organophosphate insecticides tended to be more

toxic to the larvae, whereas endosulfan was more toxic to the adults.



cotton fields. They found that all predators Coccinella sp., Paederus

aliferii (Koch.) and Metasyrphus corollae (F.) were greatly affected by

the tested insecticides except Chrysoperia carnea (Steph.), The

46


insecticides sequence programs (Decis, Cyanox, Larvin liquid and

Tamaron combi) and (Larvin liquid, Tamaron combi, Decis and Cyanox)

were the leading programs, as they induced more efficient pest control,

low efficacy on predator insects and gave high cotton yield.

Dimetry and Marei. (1992), studied the susceptibility of adults of

the cabbage aphid, Brevicoryne brassicae and two of the most important

predators, Coccinella undecimpunctata and Chrysopa carnea to

pesticides. The application of profenophos for the cabbage aphid at LC50

level exhibited no toxicity for Coccinella but was highly toxic for

Chrysopa. Pirimicarb proved to be the most selective of the compounds

tested for the control of aphids when it was used at a concentration that

caused 50 per cent mortality as it was found to be unharmful to both

Chrysopa and Coccinella. Dimethoate, on the other hand, was fto be very

harmful and highly toxic to both Coccinella and Chrysopa adults and

would probably be destructive to those natural enemies in the field.

Laboratory evaluation indicates that both pirimicarb and malathion have

potentials for use in IPM program for vegetable pest control.

El-Maghraby et al.(1993a), studied the effect of pesticidal

application during the early and late season on the abundance of certain

predators associated with cotton plants . They found that application of

kelthane-s and methomyl against mites and aphids slightly reduced the

population of Orius spp, Chrysopa carnea , Coccinella undecmpunctata

and Scymnus spp , percent reduction ranged between 10.6-17.9% also ,

they found that Orius spp was the most susceptible species tolerated the

action of the used insecticides.

El-Maghraby et al.(1993b), studied the toxicity of cypermethrin,

deltamethrin, fenpropathrin, fenvalerate, methomyl and kelthane-s against

47


the egg stage and the 1st larval instar of Chrysoperla carnea (Steph).They

revealed that deltamethrin exhibited magnitude bad side effect against the

eggs(the LC90

was 3.35 ppm) as compared with the safest compound

kelthane-s (the LC50

was 144 ppm.). The figures of the other tested

toxicants ranged between 17.6 to 109.2 ppm .They found that also the

larval stage needed higher concentration to achieve similar mortality

percentages; LC90

values ranged between 72.2 - 1176 ppm . Ranking of

toxicity was similar to that of the ovicidal action. The LC50

for the first

instar were 20.84, 18.63, 12.75, 6.32, 12.02 and 16.49 fold more tolerant

than eggs to delta-methrin, cypermeithrin, fenvalerate, fenpropathrin,

methomyl and kelthane-s, respectively. The corresponding number of

folds based on LC90

were 20.93, 20.98, 11.67, 4.83, 8.97 and 8.17 .

The selective toxicity of certain common insecticides used in

controlling aphids to the predator Coccinella undecimpuncata and its prey

cereal aphid, Rhopalosiphum spp was studied by Hussein et al. (1993a),

they reported that the contact toxicity based on LC50

values demonstrated

the selectivity of profenofos, omethoate and alphamethrin to C.

undecimpunctata with selectivity factor (S.F) of 2.75, 1.42, and 1.166,

respectively. Methmoyl showed slight selectivity (S.F 1.048). The relative

susceptibility of the predator and the aphid varied considerably among the

tested insecticides, The adult of C. undecimpunctata was more

susceptible to chlorpyrifos, malathion, pirimiphos-methyl and pirimicarb.

Fouly et al. (1995),evaluated the toxicity of Cidial and Ataboron

on the predaceous mite species Euseius scutalis (A-H.) and Agisternus

exsertus Gonzalez. The stigmaeid mite A. exsertus was more susceptible

han the phytoselid mite E. scutalis. Moreover, atabron was more effective

on both mite species.

48


Kaakeh et al.(1996),assessed the relative toxicities of two synthetic

pyrethroids (cypermethrin and fenvalerate), two organophosphorus

insecticides (chlorpyrifos, diazinon), three carbamates (propoxur,

carbaryl, bendiocarb), a phenylpyrazole representative (fipronil), and a

heterocyclic nitromethylene representative (imidacloprid or NTN 33893)

with topical bioassays in the laboratory against the convergent lady

beetle, Hippodamia convergens Guerin Meneville. They found that LD50

values decreased (i.e., toxicity increased) with an increased time after

application of a specific insecticide. The differences between the LD50

values caused by various insecticides were significant. Among tested

insecticides, cypermethrin and bendiocarb were the most toxic; fipronil

was the least toxic. H. convergens responded differently to different

insecticides within the same class. Beetles exhibited similar responses to

both organophosphorothionates chlorpyrifos and diazinon 24 to 72 h.

after application. Of the carbamates, propoxur was 2.4 and 3.5 times less

toxic than carbaryl and bendiocarb, respectively. Of the pyrethroids

tested, cypermethrin was significantly more toxic than fenvalerate. At

800 ppm, cypermethrin and bendiocarb were the fastest in killing H.

convergens among the tested insecticides. The ranking of insecticides in

decreasing order of LT50

values was as follows: fipronil > diazinon >

chlorpyrifos > propoxur > carbaryl > fenvalerate and imidacloprid.

2- Mammalian toxicity of pesticides:

Toxicity of pesticides to mammals is one of the most adverse

effects on the environment. Two main types of studies are carried out in

this respect (i. e. acute and subchronic toxicity ). Pharmacologically, the

pesticides (e.g. organophoshorus, carbamate, organochlorine and

pyrethroids) are neuropoisons which effect certain neurological targets.

49


For example, organophosphorus and carbamates esters may be classified

as anticholinesterases (Heath 1961, and O`Brien 1969) referring to their

ability to bind with AChE. However, it is known that OP compounds are

not only cholinergic agents but also they produce a variety of other

pharmacological and toxicological effects (Eto, 1947). Such effects

include those on liver and kidney functions which could be confirmed by

biochemical and histopathological studies. Many studies have been

frequently reported in this respect.

2.1. The effect on AChE and esterases.

Salem et al. (1979), studied the biochemical disorders due to

subchronic toxicity of Dursban active ingredient. They found that after

the continuous ingestion of Dursban active ingredient at dose levels of

0,40 or 80 ppm to male albino rats for three months. The poisoned

animals suffered from a successive depression in the activity of the

whole-blood AChE.

Abbassy et al. (1980), studied the toxicity from feeding

pirimphos-methyl to fayoumi laying hens. They found that no hen

mortality occurred due to treatment but blood plasma cholinesterase

activity was inhibited.

Abbassy et al. , (1981) , studied the toxicity in hens fed single

oral doses of chlorpyrifos (32 mg/kg), and pirimiphos-methyl (35 mg/kg).

These doses produced toxic effects in hens similar to that reported for

other organophosphorus insecticides. Plasma cholinesterase (ChE)

activities were significantly decreased in treated birds, but cholinesterase

activity increased after 7 days from treatment .

50


Tiefenbach and Wichner. (1985), investigated the acute action

of different doses of the organophoshorus pesticide, methamidophos on

the immune system in mice. After treatment with 1/2 and 1/10th of LD50

.

It was observed a decreasing of organ weights. There was also an

inhibition of cholinesterase in plasma and brain.

Kobayashi et al. (1986), studied the cholinergic system of brain

tissue in rats poisoned with the organophosphate, o,o-dimethyl o-(2,2-

dichlorovinyl) phosphate (DDVP),(6 mg/kg,sc , with saline as a control).

They found that actylcholinesterase (AChE)activity decreased to between

12 and 43% of the control over a period of 5 to 180 min. and recovered

almost completely within 24 hr after injection.

Sheremeta. (1986), reported that cholinesterase activity in the

blood, brain and liver of 30-day-old juveniles of Huso huso and Acipenser

stilettos exposed over 96 h to organophosphorus, organochlorine and

carbamate pesticides. Phosphamide [Dimethoate] at 1.4 and 2.1 g/liter,

2M-4X [of unstated composition] at 3.2 and 4.8 mg/liter, and benthanal

[of unstated composition] at 16.0 and 24.0 mg/litter, were found to

produce strong inhibitory effects in all tissues of both species. Primary

changes took place within as little as 3h in the blood, during the 1st day in

the liver, and during the 2nd day of poisoning in the case of brain tissue.

The results showed that total cholinesterase activity was inhibited in the

various tissues. The highest and lowest inhibitory effects were exhibited

by carbamate and organophosphorus pesticides, respectively, with

organochlorine pesticides in an intermediate position.

Khorunova and Baranova. (1987), reported that in carp exposed

to one-tenth or one-twentieth of the lethal concentration of the insecticide

51


pirimiphos-methyl for 30 days there was inhibition of serum

cholinesterase.

Singh and Drewes. (1987) studied the neurotoxic effect of low-

level chronic exposure to acephate in rats. They found that no significant

inhibition in the activity of brain AChE at doses of 1.0 or 10.0 mg/kg.

day. Low levels of Acephate exposure (1.0 mg/kg . day), which did not

alter plasma cholinesterase or RBC actylcholinesterase activity levels,

resulted in a significant elevation of plasma epinephrine and

norepinephrine levels. Decreased GABA, dopamine, and tyrosine levels

and glutamic acid decarboxylase activity in brains of these rats. Similar

changes occurred in rats exposed to 10 mg of acephate/kg day; however,

plasma chlinesterase and RBC acetylcholinesterase activities were

inhibited.

Abbassy et al. (1988), studied the side effects of profenophos and

leptophos on hen plasma. Oral administration of profenophos or

leptophos to laying hens as a single oral does, caused significant

inhibition of plasma cholinesterase (ChE). While plasma cholinesterase

activity of hens given daily small doses of each compound was

significantly reduced at the end of experiment.

Guthathakurta and Bhattacharya. (1988), treated Channa.

punctatus cultivated under paddy-cum-fish culture programs, with 176

p.p.b. phenthoate, 333 p.p.b. carbofuran, or a mixture of 88 p.p.b.

phenthoate plus 250 p.p.b. carbofuran for 15 days. The rate of mortality

was 10% in all treatments. These xenobiotics caused significant inhibition

of brain acetylcholinesterase.

Abbassy et al. (1989a), studied the effects of organophosphorus

insecticide sulprofos on serum enzymes of white rates. The acute toxicity

52


in white rats to suplrofos is moderate. The clinical biochemistry of serum

indicated significant changes the activities of µ-naphthyl esterase and

cholinesterase (ChE).

Guthathakurta and Bhattacharya. (1989), reported that in vitro

inhibition of goat cerebellar acetylcholinesterase (AChE) by pure and

commercial pesticides clearly indicates a remarkably high inhibitory

effect of commercial carbamate (e.g. carbaryl) and organophosphate (e.g.

phenthoate) pesticides containing a lower percentage of the respective

active ingredients comparable to that of the known anti cholinesterase

agents such as diisopropyl fluorophosphate and physostigmine.

Jimmerson et al. (1989) examined the inhibition of cholinesterase

(ChE) activity in the central nervous system of the rat by the potent

organophosphorus compound, soman. They found that at soman doses

greater than 55 mg/kg s.c.(0.5 LD50

), there were: (1) dose-related

inhibition of ChE activity in brain regions; (2) variability in the degree of

ChE inhibition at each soman dose in each brain region. and (3)

variability in the severity of signs of intoxication at each dose.

The chronic effects of a sublethal dose (150 mg/kg body weight)

of dimethoate on blood constituents in rats after exposure of 15 and 30

days was studied by Reena et al. (1989). They found that the activities of

acid phosphatase and cholinesterase were significantly decreased.

Lock and Johnson. (1990), studied delayed neuropathy and acute

toxicity with pirimiphos-methyl in the hen. Hens were given a single

dose of 100 mg/kg pirimiphos-methyl, which was followed by a repeated

dose after 21 days. Tri-o-cresyl phosphate (TOCP), 500 mg/kg, was used

as a positive control. All pirimiphos-methyl-treated hens received

prophylactic doses of N-methylpyridinium-2-aldoxime methanesulphate

53


(P2S) and atropine sulphate. Hens dosed with pirimiphos-methyl had very

low AChE activities (<20% of control) in both the brain and spinal cord,

24 and 48 h after dosing. In the TOCP-treated hens, the activities were

about 90% of control. All hens dosed with pirimiphos-methyl showed the

expected signs of AChE inhibition and, following recovery, usually by

day 5, no clinical signs of delayed neuropathy were seen.

Matin et al. (1990) studied the role of adrenals in diazinon-

induced changes in carbohydrate metabolism in rats. Treatment of rats

with diazinon (40 mg/kg, i.p.) resulted in reduction of brain

actylcholinesterase activity.

Sakai. (1990), studied the effect of chlorpyrifos on cholinesterase

activity in rats. given (20 mg/kg) chlorpyrifos in olive oil i.p. lowered

their plasma, erythrocyte, and brain cholinestrase activity and the plasma

activity alone was recovered within 1 month.

Yoshikawa et al. (1990), studied the effect of administration with

chlorpyrifos on electroretinograms in rats. They found that chlorpyrifos

caused abnormal ERG characterized by decreased amplitudes and

prolonged latencies of A and B waves and that the abnormal ERG did not

always correspond to the decreased retinochoroid or brain ChE activities.

Banerjee et al. (1991), studied the inhibition of human fetal brain

acetylcholinesterase. They showed that organophosphate were more

neurtoxic than carbamate compounds, as evidenced by a higher degree of

AChE inhibition by DFP and metacid-50 as compared to eserine and

carbaryl. The psychotropic durgs less potent than organophosphate and

carbamatecompounds. They showed that also pure and commercial

organophosphates and carbamates and psychotrpic drugs were all able to

significantly alter the AChE activity. Thus, the exposure of the mother to

54


these environmental toxicants may adversely affect the fetal neural

function

Cairns et al.(1991) dosed northern bobwhite, Colinus virginianus

, orally with the organophosphorus insecticide chlorpyrifos to examine

effects on brain cholinesterase (ChE) activity. They found that a lag time

of 2 to 4 h following exposure was necessary to detect significant ChE

depression caused by chlorpyrifos. The lowest dose that produced ChE

depression was between 30 and 50 mg chlorpyrifos/kg with some

mortality occurring at the high doses. Depression typically persisted for at

least 24h.

El-Harrawie et al. (1991), studied the acute oral toxicity of

methamidophos to male albino rats. Oral administration of a single dose

of LD25

of methamidophos to mature male albino rats caused significant

decrease in ChE when blood samples were taken at 1,3,6, 12, 24, 48 and

72hr after administration. On the other hand, subchronic toxicity through

administration of 50 and 100 ppm of methamidophos in drinking water

for 90 successive days resulted in a significant decrease in the plasma

ACh-ase activity.

Kandil et al. (1991) , studied the effect of subchronic doses of

cyanophos in drinking water for 12 weeks ad-libitum at two doses levels

(600 and 1200 ppm), on certain enzyme activity. It was found that the

tested doses inhibited significantly the plasma cholinesterase activity

(ChEp) throughout the experimental period.

Lavandero et al. (1991), studied the ability of fenthion to

increase gizzard erosion in broiler chicks. They found that an inverse

55


relationship between plasma cholinesterase activity and pesticide

concentration at doses higher than 1 ppm (p<0.05).

Manna. (1991), studied the influence of diazinon and fenitrothion

on acetylcholinestrase activity in digestive gland and central nervous

system of Achatine fulica. Inhibition of acetylcholinesterase (AChE)

activity in the central nervous system and digestive gland of A. fulica was

directly proportional to the concentration of diazinon and fenitrothion.

The rate of inhibition and recovery of AChE was gradual in diazinon

poisoning while it was fast in fentrothion treatment.

The effect of pesticides on chosen biochemical parameters of the

rat serum and liver was studied by Pawlowska et al. (1991). The

activities of cholinesterase (AChE), aspartate aminotransferase (GOT),

alanine aminotransferase(GPT) level were determined in the blood serum

and/or liver of rates after intragstic of oil or oil solutions of carbaryl,

methyl parathion and bromphenvinphos. They found that a distinct

increases in serum AChE and liver GOT and GPT levels and decreases in

serum GOT and GOT in rats receiving the pesticides.

Sandhu et al. (1991), studied the acute toxicity and

anticholinesterase effect of monocrotophos in domestic fowels. They

reported that monocrotophos in different doses produced marked

inhibition of erythrocyte (15.2-72.5%) and plasma (12.4-65.4%)

cholinesterase. The inhibition of blood cholinesterase was dose-dependent

and correlated well with the severity of toxic symptoms.

Siddiqui et al. (1991), studied the effects of monocrotophos and

its newly synthesized analogs, RPR-11 and RPR-V on blood chemistry

24hr. post treatment in rats given doses of 0.96, 1.23 and 3.0 mg/kg,

56


orally, respectively. Only monocrotophos caused a significant inhibition

of the brain acetylcholinesterase activity.

Anam and Maitra. (1992), studied the changed of blood glucose,

liver and muscle glycogen, and actylcholinesterase (AChE) activity in

brain and pancreas of male indian weaver brid (Ploceus philippinus

philippinus:Linnaeus) following treatment with quinalphos. Quinalphos

was given orally to birds at doses 5, 10, or 20 mg/100 g body wt./day for

10 consecutive days , respectively. They found that each dose of

quinalphos caused significant inhibition of AChE activity in brain and

pancreas.

Awal and Malik. (1992), studied the effects of daily

phosphamidon administration on certain blood biochemical

measurements in Bubalus bubalis. They found that a dose-dependent

inhibition of erythrocyte actylcholinesterase following administration of

phosphamidon to buffalo calves.

Groups of beagle dogs, were fed methidathion at constant dietary

concentration of 0-140 ppm for 1 year. The equivalent daily dosages

were approximately 0-4.7 mg/kg. There were no deaths or adverse

clinical sings associated with the treatment. Erythrocyte ChE was

inhibited in males at > 40 ppm and in females at 140 ppm. Brain ChE was

inhibited in both sexes at 140 ppm. The no- observable -effect level was 4

ppm (0.15 mg/kg/day) for ChE inhibition (Chang et al. (1992).

The effects of single acute oral doses of 1, 2.1, and 3.5 mg/kg

oxamyl (a carbamate insecticide) on selected biochemical parameters in

male Sprague-Dawley rats was studied by Fayez and Kilgore. (1992),

They found that the compound inhibited brain and blood

acetycholinesterase significantly in the first few hours of exposure. but

57


there were absence of statistically significant effects between days 7 and

14 in most of the investigated parameters.

Fossi et al. (1992), studied the serum esterase inhibition in birds:

a nondestructive biomaker to assess organophosphorus and carbamate

contamination. The birds received two single doses treatment of each

compound (azamethiphos and methomyl), i.e., 50 mg/kg and 250 mg/kg,

respectively. In the first treatment, serum butyrylcholinesterase (BChE)

and carboxylesterase (CBE) were drastically inhibited in the

azamethiphos-treated group, 24 h. after the dose. No inhibition was

detected for BChE and ChE activities in the methomyl-treated group 24 h.

after the dose. in the second treatment, the birds died or were sacrificed 3

h. after the dose. Serum BChE and brain acetylcholinestrase (AChE) were

strongly inhibited after treatment with both insecticides. Serum ChE,

hepatic microsomal CBE and 7-ethoxyresorufin dealkyation activities

were also inhibited. A statistically significant correlation between serum

BChE and brain AChE was found at lethal and sublethal doses of these

xenobiotics.

Holmes and Sundaram. (1992), studied insecticides residues and

cholinesterase inhibition in zebra finches orally dosed with fentrothion.

Dosage rates were 1.04, 3.80, and 11.36 mg/kg. Peak brain and plasma

ChE inhibitions averaged between 50-76% and 79-89%, respectively.

Plasma ChE recovered more rapidly (1-4days) than brain ChE (10 days or

more). Brain and plasma ChE inhibitions were positive, but weakly,

correlated with fenitrothion body burden.

The disposition kinetics, cytotoxicity and residues of fenvalerate

in tissues following oral administration at 5 mg/kg to goats was studied

by Mandal et al. (1992). They found that the fenvalerate did not produce

58


any significant effect on serum actylcholinesterase, cholesterol or protien

levels in goats.

Pope and Chakraborti. (1992), examined dose-related inhibition

of both brain and plasma cholinesterase activity in neonatal and adult rats

exposed to sublethal doses of three common OP pesticides, methyl

parathion, parathion, and chlorpyifos. They determined the effective dose

50 (i.e., ED50

or dose which would inhibit 50% of the cholinesterase

activity) values and then correlated them with an indicator of acute

toxicity, the maximal tolerated dose (MTD). They found that ED50

estimated for both brain and plasma cholinesterase correlated highly (r =

0.932-0.992) with previously derived MTD values. There was significant

difference between in vivo brain and plasma cholinesterase sensitivity as

expressed by ED50

values. They suggested that in vivo inhibitory potency

of the three OPs towards either brain or plasma ChE activity was highly

correlated with sensitivity to acute toxicity in both neonatal and adult rats.

Additionally, under defined experimental condition, plasma ChE

inhibition may be a useful quantitative index for the degree of brain

cholinesterase following OP exposures.

Pope et al. (1992a), compared the in-vivo cholinesterase

inhibition neonatal and adult rats by three organophosphorothioate

insecticides, (methyl parathion, parathion, chlorpyrfos.). They indicated

that neonatal rats are more sensitive to acute lethality from these

compounds and the maximum tolerated dose (MTD) exposures produce

extensive brain ChE inhibition in both age groups (7 days and 80-100

days group). They could ensure a significant inhibitor-related and age-

related differences in the duration of ChE inhibition , however, following

such OP exposures.

59


Pope et al. (1992b), studied the long-term neurochemical and

behavioral effects induced by acute chlorpyrifos treatment. They reported

that a single dose of the organophosphate insecticide chlorpyrifos (CPF),

at 279 mg/kg, s.c. caused extensive inhibition of cortical and striatal

cholinesterase (ChE) activity in adult rats at 2 (94-96%), 4 (82-83%), and

6 (58-60%) week after treatment. ChE activity was different from control

levels at 12 week after exposure.

Salama et al. (1992), investigated the pharmacokinetics and

anticholinesterase properties of a single oral dose 6 mg/kg of technical

phenamiphos in male rats. They killed the animals at time interval of 0.5-

72h after dosing . They found that the total recovered amount of

phenamiphos from brain and plasma tissues reached high level at the first

time interval and disappeared biexponentially from both to low level at

the end of the experiment. On the other hand, phenamiphos inhibited the

enzyme in both brain and plasma, where the depression of ChE activity

was usually more marked in plasma than in brain.

Vasilic et al. (1992), studied the urinary excretion rates of di-Et

phosphate and di-Et phosphorothioate and changes in blood

cholinesterase activities in fifteen persons self-poisoned either by the

organophosphorus pesticide quinalphos (twelve persons) or by

chlorpyrifos (three persons). They reported that the organophosphate

poisoning was always indicated by a significant depression of serum

cholinesterase activity in the rang of reference values took more than 30

days and had a different course in different persons.

Abdel Baki (1993), studied the detrimental effects on chickens

fed ration treated with pirimiphos-methyl. He found that the activities of

60


plasma ChE and total esterases of hens fed treated diet were significantly

inhibited within two months after treatments.

Ammar and El-Sheikh. (1993), studied the effect of confidor on

activity of some enzymes in male albino mice. They found that usage of

confidor as a single dose (1/4 LD50

, 52.5 mg/kg B.W) caused severe

inhibition of AchE after 24h of treatment. The enzyme however, started to

regain its normal level after 96h.

The compatibility of insecticides (Actellic, Reldan, Selecron and

Malathion) and chitin synthesis inhibitors CSI (IKI17899, XRD473,

Dowco 439 and SH777) and their interaction in white rats was studied by

El- Hamady (1993). The activity of plasma enzymes, cholinesterase,

total esterase, acid and alkaline phosphatases, GOT and GPT were

determined in treated rats. The levels of these enzymes in the rats treated

with the insecticides, Actellic or Malathion lonely were not significantly

different from those in rats treated with the same insecticides combined

with IKI7899 or XRD473.

Milillo et al. (1993), studied the occasional ingestion of

dimethoate by sheep. They found that when 5 sheeps were given grass

sprayed with 40% dimethoate at 4.5 litters/50 m2

, there was a fall in

serum cholinesterase values for 24h.

Osman.(1994), studied the interaction of glyphosate with some

mammalian biochemical targets. Female mice treated with 200 mg/Kg

of glyphosate as a single oral doses resulted in a maximally decreased in

AChE activity in kidney (38.9%) , liver (70.7%) and brain (75.3%)

following the single dose treatment .On other hand, following the

multiple oral dose (five doses of 200 mg/Kg glyphosate in 10 day) ,

61


recovery of AchE in brain and kidney was observed , whereas liver AChE

remained significantly.

2.2. The effect on liver function and other biochemical

parameters:

The liver is the principal organ that converts xenobiotics to

products excretable in the bile, urine and exhaled air. For these functions,

the liver has high activities of diverse enzyme systems involved in

chemical biotransformation. In particular, the liver appears to be capable

of metabolizing every chemical carcinogen, except the few which require

prior alteration by bacterial enzymes. As a consequence of its functional

activity, the liver is often the primary target for the toxic effects of

xenobiotics. Indeed even when another organ suffer the major toxic

effects of a chemical, the liver is frequently affected also, Therefore,

effects on the liver can be used as a general index for the toxicity of

xenobiotics.

The potential of xenobiotics to produce liver cell injury can often

be rapidly detected by testing up high doses. Cell injury ultimately results

in an increased permeability of the cell membrane. For liver cells, this

allows cytoplasmic enzymes such as glutamic oxalacetic transaminase

and lactic dehydrogenase to escape into the blood where their presence in

elevation of blood levels of enzymes, especially alkaline phosphatase and

g-glutamyl transpeptidase. The assessment of liver enzymes in the blood

is generally a more sensitive measure of hepatotoxicity than

histopathologic changes and can be assessed within a shorter time

(Cornelius et al, 1959 and Gradwohl,1956).

2.2.1. Transaminases.

62


Activity of certain enzymes in the blood of carp poisoned with

pirimiphos-methyl was studied by Khorunova and Baranova. (1987), in

carp exposed to one-tenth or one-twentieth of the lethal concentration of

the pirimiphos-methyl for 30 days there was an inhibition of

aminotransferase activities.

Abbassy et al. (1988), studied the effects of profenophos and

leptophos on hen plasma and liver. Single oral or daily dosing of the two

insecticides into hens significantly increased the activity of plasma GOT

and GPT than that of control hens. It was also found that the activity of

plasma GOT was always more higher than GPT in all plasma. GOT was

always more higher than GPT in all plasma samples of normal or treated

hens.

Mohamed et al. (1988), studied the degradation of Meothrin

[fenpropathrin] applied at 20 and 40 p.p.m. to barley grains during storage

for a period of up to 60 days, after which the treated grains were

incorporated into a purified diet and fed to rats for 15 days. In treated rats,

there were increases in serum aminotransferases and alkaline

phosphatase.

Ray et al. (1988), studied the effect of quinalphos (250 .mg/kg

i.p.) treatment for 13 and 26-days on the testicular steroidogenesis in rats.

They found that treatment of quinalphos for 13 days produced no toxic

effect with the exception of a significant increase in serum alkaline

phosphatase. However, after 26-days of treatment serum transaminases

and phosphatases were significantly increased.


insecticide sulprofos on serum enzymes, and liver of white rats. The

clinical biochemistry of serum indicated significant changes values in the

63


activities of glutamic pyruvic transaminase (GPT), glutamic oxaloacetic

transaminase (GOT). The results showed that the significant rise in serum

SGOT and SGPT would indicate the presence of incipient liver damage

in the initial stage as a result of exposure, to OP insecticide sulprofos.

The sublethal doses of sulprofos were administered to laying hens.

Serum enzymes, glutamic oxaloacetic acid transaminase (GOT) and

glutamic pyruvic transaminase (GPT) were measured at different intervals

from administration. Dosed hens showed significant increase of these

serum enzymes (Abbassy et al., 1989b).

Ashour et al.(1989), studied the in vivo effect of sublethal doses

of four synthetic pyrethroids, namely ; fenpropathrin, deltamethrin,

permethrin, and fenvalerate, in male and female white rats. The acute and

subchronic effect on glutamic oxaloacetic transaminase (GOT) and

glutamic pyruvic transaminase (GPT) activities in liver and brain were

determined. The four insecticides used were poinhibitors of GOT activity

in both liver and brain. The activity of GPT was decreased in brain as a

result of the insecticides treatment, whereas the enzyme level was

elevated in the liver, except with permethrin treatment as the enzyme

activity was decreased.

Reena et al. (1989), studied the hematological changes induced

by a sublethal dose (150 mg/kg body weight) of dimethoate in rats after

exposure of 15 and 30 days. The activities of glutamic-oxaloacetic

transaminase, glutamic-pyruvic transaminase and amylase markedly

increased.

Zidan et al. (1989), studied the effect of local EC. formulations of

fenvalerate and fenpropathrin pyrethroids on certain liver function

parameters in white albino mice. Mice treated by LD50

through

64


intarperitoneal injection. The tested pyrethroids when injected at LD50

levels caused no adverse effects on the parametars indicating liver

function, i.e. bilirubin, GOT and GPT enzymes systems.

Increases in blood serum concentrations of alanine and aspartate

aminotransferases were observed in rats after single intragastric dose of

o.5 LD50

of g-HCH [lindane], Bi-58 [dimethoate], Decies [deltamethrin]

or the herbicide Isophen [chlorpropham + fenuron]. This indicated liver

damage by the pesticides (Kadyrova et al., 1990).

Mohamed et al. (1990), studied the combined effect of Dursban

and Reldan on Nubian goats. They found that goats dosed orally with a

1/1 mixture of Dursban [chlorpyrifos] and Reldan [chlorpyrifos-methyl]

at 300 mg/kg body weight caused more rapid death and severe changes

than in goats dosed with insecticide alone at 150 mg/kg. Hepato-renal

lesions were accompanied by increases in aspartate aminotransferase

activity and urea concentration and by decreases in total serum protein

concentration.

Shakoori et al.(1990), studied the biochemical effects of

bifenthrin on the blood and liver of rabbit. The synthetic pyrethroid,

bifenthrin (talstar), administered to rabbits at a chronic dose of 6 mg/kg

body weight for 30 days, significantly affected the biochemical

environments of blood and liver. Activities of alkaline phosphatase

(AkP), acid phosphatase (ACP) and glutamate pyruvate transaminase

[alanine aminotransferase, AlA] decreased, respectively (11%, 22%, 48%,

33% and 18% after 15 days of insecticide treatment) while glutamate

oxaloacetate transaminase [aspartate aminotransferase, ASA] increased

92% during this exposure. On prolonged exposure for a total of 30 days,

65


the AkP activity increased 207% Correspondingly in liver, the 30-day

insecticide treatment resulted in increased AkP.



of LD25

of methamidophos to mature male albino rats resulted in a

significant elevation in the plasma GOT and GPT when blood samples

were taken at 1, 3, 6, 12, 24, 48 and 72hr after administration .

Hanafy et al. (1991), studied the biochemical and

histopathological effects of the organophosphorus insecticide, Tamaron in

rats. Groups of rats were given either a single large dose of Tamaron

(methamidophos) (20 mg/kg body wt) or repeated oral small doses (2

mg/kg body wt). Both treatments caused a highly significant elevation of

urea, aspartate and alanine aminotransferases and alkaline phosphatase in

the blood of treated rats.

Pawlowska et al. (1991), studied the effect of pesticides on

chosen biochemical parameters of the rat serum and liver. The activities

of cholinesterase (AChE), aspartate aminotransferase (GOT), alanine

aminotransferase(GPT) level were determined in the blood serum and/or

liver of rats after intragastric of oil or oil solutions of carbaryl, methyl

parathion and bromphenvinphos. They found distinct increases in serum

AChE and liver GOT and GPT levels and decreases in serum GOT and

GOT in rats receiving the pesticides

Pradhan and Dasgupta (1991), studied the effect of dimethoate

on GOT and GPT activities in plasma, liver, kidney, and heart muscle of

the male toad. The GOT activity was increased in plasma but decreased

in other organs, whereas GPT activity was decreased in plasma, liver and

66


heart but increased in kidney. The pesticide caused hepatorenal and

cardiac toxicity, resulting in alteration of transaminase activities.

Rajeev et al. (1991), studied the biochemical effects of phorate

and dimethoate, administered either orally or i.p., in female albino rats.

Phorate administration, in general, resulted in an increase in liver

transaminases and phosphatases. Significant variations were noted in

transaminases and phosphatases when dimethoate was fed.

Shiroishi et al. (1991), determined the serum enzyme activities in

12 farm dusters exposed to pesticides containing 2% fenthion, 2% O-sec-

butyl phenyl methylcarbamate and 1.5% edifenphos. The blood samples

were collected 3 times before exposure and 2-6 h and 4-6 days after

exposure. They found a positive correlation between enzyme activity

before and after dusting and blood concentration of pesticides for

glutamic-pyruvic transaminase, glutamic - oxaloacetic transaminase and

creatine phosphokinase in blood.

Siddiqui et al. (1991), studied the effects of monocrotophos and

its newly synthesized analogs, RPR-11 and RPR-V on blood chemicals .

24 hr. post treatment in rats given doses of 0.96, 1.23 and 3.0 mg/kg,

orally, respectively. The activities of membrane- bound enzymes in serum

were not significantly changed by all 3 compounds, except for a

statistically significant increase of 28% in serum GOT activity by RPR-

11.

Rao et al. (1991), studied the alterations in the rat brain

carbohydrate and related metabolism during acute and chronic acephate

toxicity. They found that activities of aspartate aminotransferase (AAT),

and alanine aminotransferase (ALAT) were decreased in activity

67


significantly during chronic treatment. However AAT, and AlAT

activities showed a significant increase during acute treatment.

Zidan et al. (1991a) , studied the biochemical effects of

Sumicidin and Sumithion, at 1,2,3,4,5 and 6 weeks after oral

administration to male mice. The results indicated clearly that Sumicidin

was a potent inhibitor of glutamic oxaloacetic transaminase (GOT) and

glutamic pyruvic transaminase (GPT) activity at 1/20 and 1/10 LD50

.

These changes were related to type of insecticide used, dose and time

elapsed.

The interaction of insecticides, Sumialpha, Sumicidin and Cyanox

with the blood plasma enzymes of male albino mice was tested by Zidan

(1991b) . Plasma glutamic oxaloacelic transaminase (GOT) and glutamic

pyruvic transaminase (GPT) activities increased depending on the dose

used and time after treatment.

Groups of beagle dogs, four/sex/dose, were fed methidathion at

constant dietary concentration of 0-140 ppm for 1 year. The equivalent

daily dosages were approximately 0-4.7 mg/kg. They found that no deaths

or adverse clinical sings were associated with the treatment. Moderate

increase in serum bile acids and enzyme activities (alanine

aminotransferase, aspartate aminotansferase, sorbitol dehydrogenase, and

alkaline phosphatase) in all dogs receiving >40 ppm (Chang et al. 1992).

Mandal et al. (1992), studied the disposition kinetics, cytotoxicity

and residues of fenvalerate in tissues following oral administration at 5

mg/kg to goats. Both GOT and GPT activities in kidney tissue, and only

GPT activities in liver tissue had decreased significantly 4, 8 and 22 days

post-administration.

68


Shakoori et al. (1992), administered a sublethal dose of karate

[lambda-cyhalothrin] to rabbits. They found that the transaminases

[aminotransferases] (glutamate oxaloacetic transaminase [aspartate

aminotransferase], GOT [aspartate aminotransferase (AST)] and GPT

[alanine aminotransferase (ALT)] increased after 15 days treatment. ALT

activity increased 119% and 60% after 15 and 30 days, respectively.

Swamy and Mohan. (1992), studied the effect of sublethal daily

dosing of monocrotophos on activities of aminotransferasand glutamate

dehydrogenase in rat brain. They found that monocrotophos caused

elevation in aspartate aminotransferase (AAT), and alanine

aminotransferase (ALAT) activities.

Tarrant et al. (1992), study the biochemical and histological

effects of the aphicide demeton-S-methyl on house sparrows (Passer

domesticus) under field condition. They found that exposure of house

sparrows to demeton-s-methyl was indicated by inhibition of serum

butyryl cholinesterase (BChE) activity and also resulted in slight

inhibition of actylcholinesterase (AChE) and raised serum glutamate-

oxaloacetate transaminase (GOT) levels in some birds.

Abedl Baki (1993), studied the detrimental effects on chickens

fed treated ration with pirimiphos-methyl. He found that plasma GPT

activity was always higher in hens fed treated ration than those of non

treated ones.

Abdel Baki et al. (1993) studied the acute toxicity of some

fungicides to rats and their effects on the structure and function of the rat

liver. Single oral doses of the tested fungicides (each equivalent to 10000

mg/kg b.w) did not exhibit any sings of toxicity symptoms but led to a

significant increase in the activity of plasma acid and alkaline

69


phosphatase and glutamic oxaloacetic transaminase (GOT) while

glutamic pyruvic transaminase (GPT) activity increased significantly in

the rats treated with the fungicide, trimeltox-forte.

Ammar and El- Sheikh. (1993), administered imidacloprid

(confidor) orally to male albino mice, using either single dose (1/4 LD50

52.5 mg/kg B.W.) or repeated doses (1/10 LD50

21 mg/kg). They found

that there was no significant inhibitory action on GOT activity in all

tested tissues, GPT level declined in both brain and kidney tissues under

various treatment conditions.

The compatibility of insecticides and chitin synthesis inhibitors

CSI and their interaction in white rats was studied by El- Hamady

(1993). The activity of plasma enzymes, cholinesterase, total esterase,

acid and alkaline phosphatases, GOT and GPT were determined in treated

rats. He found that the levels of these enzymes in rats treated with the

insecticides, actellic or malathion lonely were not significantly different

from those in rats treated with the same insecticides combined with

IKI7899 or XRD473.

Katayama.(1993), reported that feeding of 0.1% DDT containing

diet for 13-14 days to rats caused a reduction in growth rate, which was

significantly ameliorated by dietary addition of myo-inositol. Dietary

DDT increased liver weight, serum cholesterol, and activities of serum

glutamate-oxaloacetate transaminase (GOT) and glutamate - pyruvate

transaminase (GPT). The increases in liver weight, liver cholesterol,

serum cholesterol and serum GOT activity due to DDT were significantly

suppressed by dietary addition of myo-inositol. In addition, dietary myo-

inositol caused a decreasing trend in serum GPT activity in DDT-fed

animals. In the animals without receiving DDT, dietary myo-inositol

70


caused no significant effect on these metabolic parameters These results

suggest that rats fed DDT containing diet may require an exogenous

source of myo-inositol.

Male immature Wistar rats treated with a tolerated dose of (2

mg/kg/day) orally for 90 days (during the period of growth to maturity).

Twenty-four h after the termination of the treatment, organ weight and

protein concentrations were determined. Food consumption and body

weight gain decreased parallely. No changes occurred in the body tissues

but for liver which was enlarged and its protein, glutamic oxalacetic

transaminase and glutamic pyruvic transaminase concentrations increased

(Paul et al., 1993).

Interaction of glyphosate with some mammalian biochemical

targets was studied by Osman (1994). Transaminases (GPT & GOT)

activity were elevated in brain and liver while GPT or GOT activity of

kidney remained unchanged following a single oral dose (200 mg/Kg of

glyphosate). however, the multiple oral dose (200 mg/Kg of glyphosate

five times in 10 days) elevated the GPT and GOT activities of brain and

kidney, while a reduction in liver GPT and GOT activities were observed.

The acute toxicity of Ivermectin in laboratory animals studied by

Eweis et al. (1995). Three doses representing 0.5, 0.25, 0.1 of the LD50

were administered orally for 4 weeks in drinking water. The tissue-

specific enzymes, glutamate oxaloacetate transaminase (GOT), glutamate

pyruvate transaminase (GPT), the amount of urea and total protiens of

liver and kidney were significantly decreased.

2. 2. 2. Alkaline phosphatase

71


Khalifa et al. (1986), investigated the effect of determined

amounts of methyl bromfenvinphos histochemically on the liver and

spleen of hens. The liver of the treated hens has shown general increase in

the intensity of alkaline and acid phosphatases. The spleen has displayed

mild increase in the alkaline phosphatase while the acid phosphatase

activity was unaffected. It appears that the adverse changes caused by

methyl bromofenvinphos was due to its toxic effects . The toxicity has

affected the liver more than the spleen .

Khorunova and Baranova. (1987), studied the activity of certain

enzymes in the blood of carp poisoned with pirimiphos-methyl . They

found that in carp exposed to one-tenth or one-twentieth of the lethal

concentration of the insecticide for 30 days, there was an increase in

alkaline phosphatase activity.

Abbassy et al. (1988), studied the side effects of profenofos and

leptophos on hen plasma and liver. Oral administration of profenofos or

leptofos to laying hens as a single oral dose, increased the activity of

both plasma acid and alkaline phosphatases. The enzymatic activity was

significantly higher than that of the control through the experiment.

Mohamed et al. (1988), studied the degradation of Meothrin


for a period up to 60 days, after which the treated grains were


there were increases in serum aminotransferases and alkaline

phosphatase.

Ray et al. (1988), studied the effect of quinalphos at dose

250 .mg/kg i.p. for 13 and 26-days on the tesicular steroidogenesis in rats.

They found that treatment of quinalphos for 13 days produced no toxic

72


effect with the exception of a significant increase in serum alkaline

phosphatase. However, after 26-days of treatment, toxicity was

significantly increased on serum transaminases and phosphatases.


insecticide sulprofos on serum enzymes, and liver of white rats. Results

revealed that a significant increase in serum acid and alkaline

phosphatases. The significant rise in serum acid and alkaline

phosphatases indicated the presence of incipient liver damage in the

initial stage as a result of exposure to the OP insecticide.

Abbassy et al. (1989b), studied the effect of sulprofos on liver of

laying hens. Sublethal doses of Sulprofos were administered to laying

hens. Serum enzymes, glutamic oxaloacetic acid transaminase (GOT) and

glutamic pyruvic transaminase (GPT) were measured at different intervals

from administration. Sulethal doses showed significant increase of serum

enzymes.

The in vivo effect of sublethal doses of four synthetic pyrethroids,

namely : fenpropathrin, deltamethrin, permethrin, and fenvalerate, in male

and female white rats. were studied by Ashour et al.(1989). The acute

and subchronic effect on alkaline phosphatase (AP) activity in liver and

brain were determined. The enzyme activity of (AP) in liver was

increased with fenpropathrin, permethrin, and fenvalerate while it was

decreased with deltamethrin. AP activity in brain was inhibited in all

treatments.

The oxidative desulfuration of the 3 phosphorothionate

insecticides chlorpyrifos, chlorpyrifos-methyl and leptophos in rat brain

and liver was studied by Chambers and Chambers. (1989), They found

significant depletion of alkaline phosphatase.

73


The Biochemical changes induced by the insecticide fenvalerate

in the male gonads of albino rat was studied by Ghosh (1990).

Fenvalerate administration at sublethal doses (25, 50 and 100 mg/kg) for

30 days. caused significant depletion of alkaline phosphatase, resulted at

all doses.

Shakoori et al.(1990), studied the biochemical effects of

bifenthrin on the blood and liver of rabbit. The synthetic pyrethroid

bifenthrin (talstar), administered to rabbits at a chronic dose of 6 mg/kg

body weight for 30 days, significantly affected the biochemical

environments of blood and liver. Activities of alkaline phosphatase

(AkP), acid phosphatase (ACP) and glutamate pyruvate transaminase

[alanine aminotransferase, AlA] decreased to 11%, 22%, 48%, 33% and

18% of normal value after 15 days for the measured parameters of

insecticide treatments, while glutamate oxaloacetate transaminase

[aspartate aminotransferase, ASA] increased 92% during this exposure.

On prolonged exposure for a total of 30 days, the AkP activity increased

207%. Correspondingly in liver, the 30-day insecticide treatment resulted

in increased AKP, LDH [Lactic dehydrogenase], ALA activities 10%,

19%, and 40% respectively, while the hepatic ASA activity, decreased

33%.

Sivaswamy and Balachandran. (1990), studied the effect of

dimethoate on Wistar rats. Alkaline phosphatase activity increased in

liver and kidneys, and decreased in the intestines.



of LD25

resulted in a significant elevation in the plasma alkaline

phosphatase (AP), when blood samples were collected at 1,3,6, 12, 24,48

74


and 72hr after administration. On the other hand, subchronic toxicity

through administration of 50 and 100 ppm of methamidophos in drinking

water for 90 successive days resulted in a significant increase in AP

activity at both concentrations.

Hanafy et al. (1991), studied the biochemical and histological

effects of the organophosphorus insecticide, Tamaron in rats. Groups of

rats were given either a single large dose of Tamaron (20 mg/kg B W.) or

repeated oral small doses (2 mg/kg B W.). Both treatments caused highly

significant elevation of urea, GOT, GPT and alkaline phosphatase in

blood of treated rats.

Kandil et al. (1991), studied the effect of subchronic doses of

cyanophos in drinking water for 12 weeks ad-libitum at levels (600 and

1200 ppm), on certain enzymes activity. The results showed a significant

increase in the activity of plasma alkaline phosphatase (ALP) and acid

phosphatase (ACP).

Pradhan and Dasgupta (1991) estimated the acid phosphatase

(ACP) and alkaline phosphatase (ALP) activities in male toads plasma,

liver, kidney, and testis after exposure to dimethoate for period of seven

days. ACP activity increased in plasma and kidney but decreased in liver

and testis. On the other hand ALP activity increased in liver and testis but

decreased in plasma and kidney. Alteration of enzyme activities indicates

that long exposure of this systemic pesticide affects function of liver,

kidney and testis.

Rajeev et al. (1991), studied the effect of phorate and dimethoate

on growth and liver metabolism in rats. Phorate treatment generally

resulted in alterations in liver phosphatases. Animals fed dimethoate,

alkaline phosphatase decreased significantly.

75


The carcinogenic potential of dimethoate was studied by

Sivaswamy. (1991), Wistar rats treated with dimethoate (10-60 mg/kg

body weight). He found that alkaline phosphatase activity increased in the

liver and kidneys, but declined in the intestines.

Sandhu et al. (1992), found that single oral doses of 20 and 40

mg/kg of monocrotophos insecticide caused almost complete depletion of

fine granular alkaline phosphatase (AKPase) activity in the buffalo

hepatocytes.

Effect of prolonged administration of insecticide (cyhalothrin/

karate) was studied by Shakoori et al. (1992), They found hepatic

alkaline phosphatase activity decreased 30% after 15 and 30 days.

Abdel Baki (1993), studied the detrimental effects on chickens

fed pirmiphos-methyl treated ration. He found that the activities of plasma

acid and alkaline phosphatases were significantly increased in the hens

fed pirmiphos- methyl- treated ration stored up to 3 months from

treatment.

Ammar and El-Sheikh. (1993), studied the effect of Confidor on

activity of some enzymes in male albino mice. They administered

imidacloprid (Confidor) orally to male albino mice, using either single

dose (1/4 LD50

52.5 mg/kg B.W.) or repeated doses (1/10 LD50

21

mg/kg). ALKpase enzyme activity was reduced greatly in all tested

tissues under different dosing treatments. Though after 24h post

treatment, its level increased significantly in both liver, and kidney

tissues.

2. 2. 3. Cholesterol, bilirubin, total protien and albumin

76


El-Harrawie et al. (1986), studied the toxicity of fenvalerate and

methomyl to male albino mice. Oral administration of doses

corresponding to 1/4 or 1/2LD50

from either insecticide caused an

increase in the serum total protein level.

Saleh et al. (1986), studied the comparative toxicity of

flucythrinate and fenvalerate to albino rats. Fenvalerate and their

commercial formulations 30% EC and 20% EC, respectively, were tested

for their subchronic dermal toxicity to albino rats at different

concentrations. Either flucythrinate or fenvalerate caused a significant

reduction in serum protien level after 2 weeks of the treatment.

Mohamed et al. (1988), studied the degradation profile and

residual toxicity of meothrin during storage of barley grains. Meothrin


for a period up to 60 days, after which the treated grains were


there were reductions in serum albumin and total proteins, increases in

serum bilirubin and kidney function was also affected.

The sub lethal effects of baygon [propoxur] up to 0.375% and

nuvan [dichlorvos] up to 0.1% on the tadpoles of Rana.

hexadactyla(Lesson) was studied by Raj et al. (1988), They found that

total body protein and liver glycogen were reduced in all of the pesticide-

exposed tadpoles.

Ray et al. (1988), studied the effect of quinalphos on testicular

steroidogenesis in rats. They observed an increased cholesterol level in

testis rats treated by 250 mg/kg i.p quinalphos for 13 day.

77


Zidan et al. (1988), studied the effect of ready and locally

prepared formulation of fenvalerate and fenprothrin pyrethorids on the

acute toxicity, haematological and some biochemical changes in blood of

albino mice after interapritoneal injection by the median lethal dosage.

Results indicated that the tested formulations when used at LD50

by

intraperitoneally injection caused no adverse effect on protien contents of

male mice plasma.

Rao. (1989), studied the phenthoate impact on the ammonia and

nitrogen metabolism in the body fluid of the snail, Pila globosa

(Swainson). They found that treatment of the snail Pila globosa with

phenthoate at lethal and sub lethal rates resulted in a reduction in total

proteins.

Reena et al. (1989), studied the hematological changes induced

by a sublethal dose (150 mg/kg body weight) of dimethoate in rats after

exposure of 15 and 30 days. After 30 days of exposure, the level of

cholesterol markedly increased. But there was no effect on total plasma

protien content.

Ali and Shakoori.(1990), fed aldrin-mixed diet to Sprague

Dawley rats at a dose of 20 mg, 8 mg and 2.5 mg/kg body wt./day for 48

hours, 15 days, and 18months, respectively . The certain phosphokinase

activity increased (83%) in 15 day treatment. Bilirubin content increased

also in 15 day.

Kadyrova et al.(1990), study the effect of acute pesticide

poisoning on lipid, protien, and carbohydrate metabolism in experimental

animals. They found that an increase in blood serum concentrations of

cholesterol in rats after single intragastric dose of o.5 LD50

of g-HCH

78


[lindane], Bi-58 [dimethoate], Decies [deltamethrin] or the herbicide

Isophen [chlorpropham + fenuron].

Saleh. (1990) studied the effect of oral administration of

methomyl on protein metabolism in male albino rats after 1, 2, 3, and 4

weeks of treatment. Serum total proteins, albumin, albumin/globulin's

ratio were significantly decreased while urea and uric acid were increased

in a cumulatively dose-related manner. There were significant decreases

in serum total globulins and creatinine levels which were only evident in

the third and fourth week respectively. These results were discussed in

relation to liver and kidney functions.

The biochemical effects of bifenthrin on the blood and liver of

rabbit was studied by Shakoori et al.(1990). The synthetic pyrethroid

bifenthrin (Talstar) was administered to rabbits at a chronic dose of 6

mg/kg body weight for 30 days, on prolonged exposure for a total of 30

days, concentration of glucose, cholesterol, bilirubin and protein

decreased 18%, 19%, 35% and 34%, respectively. Correspondingly in

liver, the 30-day insecticide treatment resulted in concentration of

glycogen and protein decreased 60% and 12% respectively.

Sivaswamy and Balachandran. (1990), studied the effect of

dimethoate on Wistar rats. There were appreciable changs in urea and

protein levels .

Zaidi et al.(1990), reported that the levels of total lipids,

phospholipids, cholesterol were estimated in the cerebral hemisphere,

cerebellum and brain stem of rats administered with monocrotophos at 2,

3 and 4 mg/kg body weight i.p. daily for 10 days. All levels were

depleted following this treatment.

79


El-Harrawi et al. (1991), studied the acute and subchronic

toxicity of methamidophos to male albino rats. Oral administration of a

single dose of LD25

of methamidophos to mature male albino rats resulted

in a significant elevation in the plasma total protein at 1, 3, 6, 12, 48, and

72hr after administration. On the other hand, subchronic toxicity through

administration of 50 and 100 ppm of methamidophos in drinking water

for 90 successive days resulted in a significant increase in total protien at

both concentrations in biweekly samples.

Vandana et al. (1991), studied the biochemical effects of phorate

and dimethoate in female albino rats. They found that liver proteins

decreased with both pesticides.

Zidan (1991), determined the biochemical damages in male white

albino mice after intrapretoneal injection with LD50

and 1/2LD50

of

Sumicidin and Sumithion. Sumicidin increased total protein 48hr.

following treatment while Sumithion decreased it.

The biochemical effects of Sumicidin and Sumithion, at 1, 2, 3, 4,

5 and 6 weeks after oral administration to male mice were studied by

Zidan et al. (1991a),. Sumicidin at 1/20 LD50

( 32.78 mg/kg ) caused

increased of total protien after oral application at 4, 5 and 6 weeks

recording 116.7, 121.21, and 106.25 %, respectively. Where Sumithion

was more effective on total protien.

Zidan et al. (1991b), studied the effect of Sumi-alpha, Sumicidin

and Cyanox insecticides to male white albino mice on total protein and

total free amino acids. All pesticides at LD50

decreased total protein after

4 hours from treatment recording 47.50, 57.78, and 51.22 % of control on

intestine, respectively. This effect was more marked at LD100

of Sumi-

80


alpha and Sumicidin was used recording 62.50, and 62.55%, respectively,

maximum effect was shown by dosing LD100

of Sumi-alpha and Cyanox

recording 98.86 and 100 % respectively. The obtained results revealed

decreased in total protien according to the type of insecticide and the

dosage used.

Awal and Mailk. (1992), studied the effects of daily

phosphamidon administration on certain blood biochemical

measurements in Bubalus bubalis. Phosphamidon exerted less

pronounced effect on blood glucose and total serum protien.

The effects of single acute oral doses of 1, 2.1, and 3.5 mg/kg

oxamyl (a carbamate insecticide) on selected biochemical parameters in

male Sprague-Dawley rats were investigated by Fayez and Kilgore

(1992), They found significant changes in serum total lipids and glucose

when oxamyl was given at 2.1 and 3.5 mg/kg, but serum protein was not

affected at any dose level.



by Mandal et al. (1992). They found that fenvalerate did not produce

any significant effect on serum actylcholinesterase, cholesterol or protien

levels in goats.

Shakoori et al. (1992), studied the effect of prolonged

administration of insecticide (cyhalothrin / karate) to rabbits. They found

that the glucose content increased 17% and 185%, while cholesterol

decreased 40% and 66%, and bilirubin 84% and 61% after 15 and 30

days, respectively.

81


Agrawal and Sultana. (1993), observed the changes in

biochemical constituents of rat erythrocyte membranes after a single ip

exposure to 300mg commercial HCH /kg body weight (one-third of the

LD50

). The phospholipid: cholesterol ratio was altered. The phospholipid

content was increased while cholesterol was significantly decreased.

Eissa and El-Sheikh. (1993), studied the genotoxicity of the

systemic insecticide Confidor in male albino mice. They indicated that

toxic effects on genetic material (DNA and RNA) and total protien

(albumin and globulin), regardless of dose quantity or duration post

treatment.

Male immature Wistar rats treated with a tolerated dose of 2

mg/kg/day endosulfan orally for 90 days (during the period of growth to

maturity). Twenty-four h. after the termination of the treatment, food

consumption and body weight gain were decreased parallely. No changes

occurred in the body tissues but for liver, it was enlarged and its protein,

glutamic oxalacetic transaminase and glutamic pyruvic transaminase

concentrations were increased.(Paul et al; 1993).

The changes of some serum parameters and amino acids content

in rats after chronic sub lethal doses of dimethoate was studied by

Hassan et al. (1994) An intraperitoneal repeated doses of dimethoate

(O,O-dimethyl S-(N-methyl carbamoyl methyl) phosphorodithioate) was

injected to male Wistar rats for 8 successive days. Body weight and liver

weight decreased significantly while liver/body weight ratio and mg

protein/g liver remained unchanged.

The acute toxicity of Ivermectin in laboratory animals was studied

by Eweis et al. (1995). Three doses representing 0.5, 0.25, 0.1 of the

82


LD50

were administered orally for 4 weeks in drinking water. The tissue-

specific enzymes glutamate oxaloacetate transaminase (GOT), glutamate

pyruvate transaminase (GPT), the amount of urea and total protiens of

liver and kidney were significantly decreased.

2.3. Kidney function:

The kidney of mammals is an extremely complex organ, both

anatomically and functionally. One primary renal function is execration

of wastes, but the kidney also plays a significant role in the regulation of

total body homeostasis . The kidney is the predominant organ involved in

regulation of extracellular volume and in controlling electrolyte and acid-

base balance. This organ is also the major site of formation of hormones

that influence systemic metabolic function. A toxicological insult to

kidney could affect any or all of these functions. However, the effects

usually reported following toxic insult reflect decreased elimination of

wastes, i.e. an increase in blood urea nitrogen (BUN) or an increase in

plasma creatinine (Punia et al. 1987 ; Dheranetra et al. 1988 ; Zidan et

al. 1989 and Ogata and Izushi, 1991).

Hamza et al. (1981), studied the toxicity of fenpropathrin in dogs.

The formulated product of this insecticide was administered twice weekly

fa period of 10 weeks. Biochemical changes of these dogs during the

course of toxicity were investigated. The picture of blood urea nitrogen in

control group ranged from 25 and 26 mg/100ml blood. There were no

changing the treated group although the histopathological picture showed

degenerative change in the kidneys.

Saleh et al. (1986), studied the biochemical disorders attributable

to repeated dermal application of two synthetic pyrethroids, flucythrinate

83


and fenvalerate. Each of the insecticides caused elevation in both blood

urea and blood glucose levels after 2weeks of treatment.

Punia et al.(1987), studied the effect of phenthoate on

haematological and biochemical parameters in young male rats following

the administration of 12.5 and 25 mg/kg i.p. daily for 28 days. They

observed a significant increase in blood creatinine level by day 28 at 25

mg/kg.

Dheranetra et al. (1988). administered a single oral dose (20-40

mg/kg) of O,S,S-trimethyl phosphorodithioate, a malathion impurity with

delayed toxic properties, to rats. They observed that blood urea

nitrogen/creatinine ratios were less than 15:1. The effect of oral

administration of methomyl on protein metabolism in male albino rats

after 1, 2, 3, and 4 weeks of treatment was studied by Saleh (1990).

Serum total proteins, albumin, albumin/globulin's ratio were significantly

decreased while urea and uric acid increased in a cumulatively dose-

related manner. There were significant decreases in serum total globulins

and creatinine levels which were only evident in the third and fourth week

respectively. These results were discussed in relation to liver and kidney

functions.

Rabbits exposed to prolonged exposure at dose 6 mg/kg of the

pyrethriod bifenthrin for a total of 30 days showed increase in the

concentration of urea (Shakoori et al. , 1990)

Hanafy et al. (1991), studied the biochemical effects of OP

insecticide, Tamaron in rats. Rats were given either a single large dose of

Tamaron (20 mg/kg.) or repeated oral small doses (2 mg/kg.). Both

treatment caused a highly significant elevation of blood urea.

84


Rats receiving abamectin in drinking water at concentration 63 or

90 ppm for two months showed hepatic and renal toxicity as indicated by

the significant elevation of certain biochemical parameters in serum (i. e.

alkaline phosphatase, aspartate aminotransferase, alanine

aminotransferase, total bilirubin, creatinine, urea and uric acid (El-

Hamady,1997).

2.4. The effect on body weight:

The effect of pesticides on body weight is important when

discussing the toxicity to mammals as it reflects the general physiological

and hygienic state of the organism. Several studies were performed in this

respect.

Abbassy et al. (1981) , studied the toxicity and residues in hens

fed single oral doses of chlorpyrifos (32 mg/kg), and pirimiphos-methyl

(35 mg/kg). These doses produced toxic effects in hens similar to that

reported for other organophosphorus insecticides. Feed consumption,

body weight gain, egg production, egg weight and shell thickness were

greatly reduced. The livers of treated chickens were normal, while the

ovaries were significantly lighter than those of the control group.

Lapadula et al. (1985), studied characterization of delayed

neurotoxicity in the mouse following chronic oral administration of Tri-o-

cresyl phosphate. They found that a daily dosing of 225 mg/kg TOCP for

270 days caused a decrease in body weight gain.

The acute action of different doses of the organophoshorus

pesticide methamidophos on the immune system in mice was studied by

Tiefenbach and Wichner (1985). They found that after treatment with

1/2 and 1/10th of LD50

there was observed decreas of organ weights.

85


El-Harrawie et al. (1986), studied the toxicity of fenvalerate and

methomyl to albino mice. Oral administration of doses corresponding to

3/4 or 1/2 the LD50

of either insecticide caused reduction of body weight.

Ray et al. (1988), studied the effect of quinalphos (250 .m.g/kg

i.p.) treatment for 13 and 26-days on the testicular steroidogenesis in rats.

They found that treatment of quinalphos for 13 days failed to produce any

effect on the relative weights of testes and accessory sex glands. But the

rats treated for 26 days showed a reduction in the relative weights of

testes and accessory sex glands.

Pillai et al. (1989), studied the effect of subacute levels of Naled

on glutathione levels in liver and kidneys of rats. Rats were given 12.5,

25.0 and 50.0 mg Naled in food for 90 days. They observed a decrease in

body weight gain in male rats fed Naled. Female rats did not show any

change in body weight gain.

Agarwal et al. (1990), studied the effect of sub acute insecticide

exposure in male albino mice treated with phosphamidon, propoxur or

aldrin at 1/40 LD50

that was dosed intraperitoneally daily for 8 weeks.

Body weight was not affected by the insecticides.

El-Gendy. (1991), studied the effect of daily oral administration

of sub-lethal doses of lindane (5 mg/kg) and deltamethrin (2 mg/kg) for

three weeks on male mice after one hour and after three weeks of last

dose. The treated mice showed reduction of body weight. There was also

a significant increase in liver, kidney and brain : body weight ratio after

three weeks of last dose. In contrast, there was a significant decrease in

spleen : body weight ratio.

86


Effects of chlordane on parameters of liver and muscle toxicity in

man and experimental animals was studied by Ogata and Izushi(1991).

In a simultaneous study, rats were administered 100 mg/kg body wt. of

chlordane by stomach tube once a day for 4 days, whereas 50 mg/kg body

wt. of chlordane was injected intraperitoneally once a day for 4 days. The

data showed that a significant increase in liver weight,

The biochemical effects of phorate and dimethoate, administered

either orally or i.p., in female albino rats were studied by Rajeev et al.

(1991). They found that there was an increase in body weight in phorate-

fed rats and a decrease in body weight in dimethoate-fed rats. They

observed also an increase in liver, kidney, spleen and brain weights in rats

fed with phorate and dimethoate.

Sivaswamy (1991), studied the carcinogenic potential of

dimethoate in wistar rats treated with dimethoate (10-60 mg/kg body

weight).They found an increase in weight over a 3-month period.

Groups of beagle dogs, were fed methidathion at constant dietary

concentration of 0-140 ppm for 1 year. The equivalent were

approximately 0-4.7 mg/kg. There were no deaths or adverse clinical

sings associated with doses in treatments. Weekly body weights and

weight gain were not affected (Chang et al. 1992).

Fayez and Kilgore. (1992), studied the effects of single acute oral

doses of 1, 2.1, and 3.5 mg/kg of oxamyl on selected biochemical

parameters in male Sprague-Dawley rats. They found that the animals

exhibited significantly decreased weight gain when compared to control

animals.

87


Feeding of 0.1% DDT containing diet for 13-14 days to rats

caused a reduction in growth rate, which was significantly ameliorated by

dietary addition of myo-inositol. Dietary DDT increased liver weight. The

increases in liver weight due to DDT were significantly suppressed by

dietary addition of myo-inositol. In the animals without receiving DDT,

dietary myo-inositol caused no significant effect Katayama,(1993).

Male immature Wistar rats were treated with a tolerated dose of (2

mg/kg/day) of endosulfan orally for 90 days (during the period of growth

to maturity). Twenty-four h after the termination of the treatment, organ

weight, was determined. Food consumption and body weight gain were

decreased parallely. No changes occurred in the body tissues but for liver

which was enlarged and its protein, glutamic oxalacetic transaminase and

glutamic pyruvic transaminase concentrations increased (Paul et

al.,1993).

The influence of bromfenvinphos alone, and in mixture with

methoxychlor, on levels of gamma-glutamyl transpeptidase,

ceruloplasmin and cholesterol in the blood plasma of laboratory mice was

studied by Zaleska and Wolewicz (1993). Bromfenvinphos (BrV), 12.33

mg/kg/day alone and in combination with methoxychlor(MeOCl), at

24.66 mg/kg/day were administered daily (intragastrically) in pure olive-

oil for 6 weeks to male laboratory mice. The relative weight of liver was

higher in mice receiving both drugs in combination, than in control and in

mice BrV alone. The relative weight of spleen was significantly higher in

animals receiving BrV alone, than in other groups.

Hassan et al. (1994), studied the changes of some serum

parameters and amino acids content in rats after chronic exposure to sub

lethal doses of dimethoate. An intraperitoneal repeated doses of

88


dimethoate [(O,O-dimethyl S-(N-methylcarbamoyl methyl)

phosphorodithioate)] was injected to male Wistar rats for 8 successive

days. Body weight and liver weight were decreased significantly while

liver/body weight ratio and mg protein/g liver remained unchanged.

Abdel-Nasser.(1995), evaluated the immunotoxic effect of

diazinon , carbofuran and cypermethrin in female mice. Mice was dosed

by giving with 0.01 , 0.001 and 0.0001 of the oral LD50

of each technical

insecticide .No changes in body weight except with high dose of

cypermethrin. Reduction in spleen celluiarity and weight of intoxicated

mice were found.

Attia (1995),studied the effect of dimethoate on rat pineal and

serum melatonin and hepatic glutathion levels. He reported that

lowerbody weight gain on male albino rats was due to the effect of

dimethoate (5 and 10 mg/Kg/day oral dose for 6 successive days) .

Eweis et al. (1995), studied the acute toxicity of Ivermectin in

laboratory animals. Body weight gain and food consumption were

decreased clearly at 0.5 LD50

of Ivermectin. In contrast organs

weight/body weight ratio was higher than that observed in non treated

animals.

Danielson and Golsteyn(1997) studied the body weight and food

consumption in Hereford steers, under feedlot conditions, for 10 week

after exposure to diazinon. Daily weight gain and feed utilization in

treated animals with diazinon, during the 70-days study, were not

altered.

2. 5. Histopathological studies:

89


Khalifa et al. (1986), investigated the effect of determined

amounts of methylbromfenvinphos histochemically on the liver and

spleen of hens. Histological examination of the liver showed cellular

hypertrophy, intraytoplsmic vaculations and sinusoidal dilations. Spleen

also showed a loss of the clear distinction between the red and white

pulps. Mean time the white pulps are found more disrupted. The red

puple were more dilated and engorged with blood.

Gajdova et al. (1988) studied the oestrogenic effects of the

organophosphate pesticide phosmet on the uterus of neonatal rats.

Compared to controls, the relative weight of the uterus was significantly

increased after administration of 1/25 and 1/50 LD50

of phosmet and of

diethylstilbestrol dipropionate. Histological examination of the uterus

revealed changes indicative of oestrogenic stimulation of the uterus.

Abbassy et al. (1989), studied The effect of sulprofos on liver of

laying hens. Sublethal doses of sulprofos were administered to laying

hens. Liver of sulprofos-treated hens revealed marked degeneration and

fatty change in the hepatic cells.

Hanafy et al. (1991), gave groups of rats either a single large dose

of Tamaron (methamidophos) (20 mg/kg body wt) or repeated oral small

doses (2 mg/kg body wt). They found that both treatments caused

degenerative and vascular changes in the liver, kidney, heart, testis and

brain of rats.

Ogata and Izushi (1991),compared the toxic effects of the

organochlorine insecticide 'chlordane' in man and rats. They administered

100 mg/kg body wt. of chlordane by stomach tube once a day for 4 days,

or injected intraperitoneally 50 mg/kg body wt. of chlordane once a day

90


for 4 days to rats. They reported that histological examination of the liver

confirmed fatty infiltration induced by chlordane in rats.

Sahai et al. (1991), studied the effect of sublethal doses (1 ml/kg)

of Malathion administered every fifth day for 30 days on hepatic cells of

rats. They reported that hypertrophy in hepatic cells and their nuclei and

in intralobular vein etc. were observed.

Shakoori et al. (1992), reported that administered of sublethal

dose of karate [lambda-cyhalothrin] to rabbits produced a histological

changes, were marked by atrophied hepatic cells, and hypertrophied

nuclei and nucleoli. also a trend towards necrosis of hepatic cells was

observed.



by Mandal et al. (1992). They found that fatty changes in periphery of

lobule, congestion in sinusoid, haemolysis in central vein, necrosis and

periphery fibrosis around the central vein of liver, and necrosis in kidney

of fenvalerate treated goats.

Agrawal and Sultana. (1993), studied the changes in

biochemical constituents of rat erythrocyte membranes after a single ip

exposure to 300 mg commercial HCH /kg body weight (one-third of the

LD50

). They reported that the erythrocytes showed morphological

change(cell deformity and echinocyte formation.).

Baronia and Sahai (1993), studied the effects of sublethal dose

(500 mg/kg body weight) of DDT on the testis of albino rat up to six

weeks. They reported that necrosis in seminiferous tubules, hypertrophy

91


in germinal epithelium and sperms, shrinkage of sertoli cells,

spermatogonia, spermatocytes and spermatids.

Chowdhury et al. (1993) reported that intraperitoneal treatment

with lindane in male mature rats weighing 150±5g at doses of 4 and 8 mg/kg

caused marked regression in cellular and pharmacological properties of accessory organs. The

decrease of weight in accessory organs was conspicuous after lindane treatment. Morphological

degeneration of accessory organs (epididymis, seminal vesicle, vas deferens) was markedly evident

with 8 mg/kg lindane treated group. Contractile property of vas deferens and seminal vesicle was also

inhibited by in vitro lindane treatment. Morphological regression in male accessory organs and

inhibition of contractile property were in agreement and thus shows the adverse effects of lindane.

92

MATERIALSMATERIALS AND AND

METHODSMETHODS

MATERIALS AND METHODS

1- Test insects:

1.1. The cotton leafworm: Spodoptera littoralis (Boisd)

Egg - masses of the cotton leafworm Spodoptera littoralis

(Lepidoptera: Agrotidae) were brought from the laboratory of Sakha

Agricultural Research station. Rearing of this strain was continued in the

laboratory as described by El - defrawi et al.(1964). The method

consisted of rearing the larvae on castor oil-bean leaves Ricinus

communis under conditions of 25° C and 70% relative humidity. Three

days old larvae were transferred to large glass containers, covered with

muslin secured with rubber bands, and supplied daily with fresh castor

bean leaves for feeding.The number of larvae per jar were adjusted

according to the larval instar. 300, 100, 30 and 15 for the frist, second,

third and fourth instars, respectivily, while, 10 larvae were used for each

fifth and sixth instar. Five to ten of the survived sixth instar larvae were

transferred to clean jars containing sow - dust for pupation. Pupae were

transferred to petridishes and kept in cages for the emergence of adults

which were then supplied with leaves of Nerium oleander as a suitable

site for oviposition and 10% sugar solution for feeding. The egg hatches

were daily collected and reared as mentioned before. The fourth instar

larvae were used for laboratory experiments.

1.2.Aphids:

The adults stages of aphids { Aphis gossypii (Glover), Aphis

craccivora(Kock) and Brevicoryne brassicae(L.) [Homoptera:

Aleyrodidae]}, were obtained from okra, beans, and cabbage plants,

respectively from Kafr - El-Sheikh Governorate.Cultures were maintained


under laboratory conditions on suitable plants for six months free from

pesticidel contamination. Kenaf Hibiscus cannabinus, broad bean Vicia

faba and cabbage Brassica oleracea var capitata seedlings were used for

rearing aphids (Aphis gossypii, Aphis craccivora and Brevicoryne

brassicae, respectively. Aphids were always transferred weekly or after

two weeks from old to young seedlings by cutting the heavily infested

leaves and were placed on the new plants. Contamination between

cultures was prevented by placing these seedlings in special chambers

50x50x60 cm. covered on their sides with muslin. These cultures were

maintained in a breeding room under the temperature 25±2°C and 65±5

R.H and 12 hours daily illumination by 2 fluorescent bulbs of 40 wts.

each.

1.3. Spider mite Tetranychus cinnabarinus (boisduval):

Spider mite T. cinnabarinus (Acarina:Tetranychidae) colonies were

obtained from castor bean plants from Kafr El-Sheikh Governorate and

reared under laboratory conditions on castor bean Ricinus communis (L.)

for about six months away from any contamination of pesticides before

starting the experiments. About 6-10 seeds of castor bean were planted in

one pot (30 cm.diameter) and left under the green house conditions for 7-

10 days for germination. After 7-10 days, the seedlings were infested by

clean culture of red mites. Mites were always transferred from old to

young plants by cutting heavily infested leaves into small sections which

were then placed on new plants. Contamination was prevented by placing

these seedlings in special chambers 50x50x60 cm. covered with muslin.

These cultures were maintained in a breeding room under the temperature

25±2° C and 65±5 RH and 12 hours daily illumination by 2 fluorescent

bulbs of 40 wts each. Mites were collected by placing the infested castor-

96


oil bean leaves on white paper, then the full mature individuals were

chosen and transferred by a fine brush to discs for treatements.

1.4. The predator, paederus alfierii (Kock):

The tested predator P. alfierii (Staphylinidae: Coleoptera) was

collected from untreated vegetable fields in Kafr - EL-Sheikh

Governorate by using an insect trap and was transferred to the laboratory.

Predators were placed in glass jars each of one littre covered with muslin.

New preys ( aphis sp, eggs of Spodoptera littorals) were offered every

day to the predator to keep a constant supply of food. Predators were kept

under laboratory conditions ( temperature 25±2 °C and 65±5 RH and 12

hours daily illumination by fluorescent light) for at least 2 weeks before

testing.

2-Test animals:

A pure strain of healthy white albino male and female rats were

purchased from Faculty of Medicine, Tanta Univ., and reared in the

laboratory .The animals were housed in metallic cages, provided ad

libitum with balanced ration consisted of bread, carrot and milk. Male rats

of 3-4 months age and 150-180 gm weight were chosen for the

experiments.

3-Pesticides

Trade name :Marshal.

Common name :Carbosulfan.

Mol. Formula :C20H32N2O3S.

Chemical name :2,3-dihydro -2,2-dimethyl benzofuran -7-yl

(dibutylaminothio) methyl carbamate.

Structural formula

97


O

OCON

CH3

S N[(CH2)3CH3]2

CH3

CH3

Formulation : W.P 25 %

Introduced by :FMC Corparation Agriculfural Chemical Group.

Philadlefhia, Pennsylvania.

Trade name :Pirimor.

Common name :Pirimicarb.

Mol. Formula :C11H18N4O2.

Chemical name :2-dimethyl amino-5,6-dimethyl pyrimidin-4-yl

dimethyl carbamate.

Structural formula :

N

N

OCON(CH3)2

CH3

CH3

N(CH3)2

Formulation :WP 50 %.

Introduced by :ICI plant protection Division. England.

Trade name : Unden.

Common name :Propoxur.

Mol. Formula :C11H15N03

Chemical name :2-isopropoxyphenyl methyl carbamate.


98


OCONHCH3

OCH(CH3)2

Formulation :EC 50 %.

Introduced by :Bayer, Agrochemical Division.Leverkusen, Germany.

Trade name :Reldan.

Common name :Chlorpyrifos-methyl.

Mol. Formula :C7H7Cl3NO3PS

Chemical name :O,O-dimethyl O-3,5,6-trichloro-2-pyridyl

phosphorothioate.


NCl

Cl Cl

OP(OCH3)2

S

Formulation :EC 50 %

Introduced by :DOW. Elanco, USA.

Trade name : Dimethoate.

Common name :Dimethoate.

Mol. Formula :C5H12NO3PS2

Chemical name :O,O-dimethyl S-methyl carbamoyl methyl

phosphorodithioate .


99


CH3NHCOCH2SP(OCH3)2

S


Introduced by :Kafr El - Zayat Company.

Trade name :Cidial.

Common name :Phenthoate.

Mol. Formula :C12H17O4PS2.

Chemical name :S-ethoxy carbonyl benzyl O,O-dimethyl

phosphorodithioate.


CHSP(OCH3)2

CO2CH2CH3

S

Formulation :EC 50 %

Introduced by :ISAGRO - MILANO - Italy.

Trade name :Actellic.

Common name :Pirimiphos-methyl.

Mol. Formula :C11H20N3O3PS

Chemical name :O-2-diethyl amino -6-methylpyrimidin -4-yl O,O-

dimethyl phosphorothioate


100


N

NCH3

OP(OCH3)2

N(CH2CH3)2

S


Introduced by :ICI plant protection Division. England.

Trade name :Selecron.

Common name : Profenofos.

Mol. Formula :C11H15BrClO3PS.

Chemical name : O-4-bromo-2-chlorophenyl O-ethyl S-propyl

phosphorothioate.


O

SCH2CH2CH3

Br

Cl

OCH2CH3P

Cl

O

Formulation : EC 72 %.

Introduced by : CIBA. GEIGY Limited. Basle, Switzerland.

4-Laboratory-toxicity tests:-

4.1. The cotton leaf worm, Spodoptera littoralis :-

The leaf dipping technique was used. Serial concentrations of the

tested pesticides were prepared in water. Leaves of castor bean were

immersed for 2 seconds in the prepared concentrations. Leaves were left

for dryness. Ten of fourth instar larvae were allowed to feed on the

101


treated leaves for 24 hrs. Three replicates were made for each

concentration. Mortality counts were recorded.

4.2. Aphids

The slide dipping technique described by El- Sayed et al. (1978) was

applied to assay the toxicity of different pesticides against aphids. A piece

of double faced scotch tape was pressed tightly to the surface of a glass

slide. Using a moist brush, ten adults (1-2 days old) were stuk to the tape

on their backs so that thier legs and antennae were free. The infested

slides were then dipped in the pesticides dilutions and gently agitated for

five seconds. Any excess of the solutions was removed using a filter

paper and kept under the same conditions of the breeding room. Four

replicates were used for each concentration. Forty of insects was also

dipped in water according to the same technique and considered as

control check. Mortality counts were recorded 24 hours following

treatments. Aphids responding to touch of a fine brush were considered

alive.

4.3. Mites :

A leaf-disc dipping method described by Abo EL -Ghar and El-

Rafie (1961) was applied to evaluate the susceptibility of adult female

mites to different pesticides. Four castor bean leaf discs (1.5 cm in

diameter) were placed upside down on a filter paper over a wet cotton

pad in petri-dish, 9 cm in diameter. Treatments were carried out by

immersing the leaf disc in the tested pesticidal dilution for five seconds,

and the treated discs were left to dry and returend to the petri-dishes. Ten

adult mites were placed on the exposed surface of each disc and kept

under the same conditions of breeding room. Each dish contained four

discs and this was replicated 4 times. Using microscopical examination,

102


mortality was counted after 24 hours, and mites responding to touch of

brush were considered alive.

4.4. The predator, P. alfierii :

Surface deposit technique was used to determine the toxicity of

pesticides to the predator as described by Mostafa et al (1980). The dry

film of pesticides was prepared by applying 1 ml. acetone containing the

desired concentration of each pesticide on 9 cm. diameter petri dish, after

the solvent was evaporated, ten adults of the predators were transferred to

treated petri dishes and this was replicated 4 times. Prior to use, the

predators were exposed to low temperature ( 4 °C for 10 min.). Mortality

counts were recorded after 24 hours from treatment.

4.5. L.D.P lines and statistical analysis:

For all insects, mortality percentages were corrected according to

Abbott`s formula (1925), and plotted on probit graph paper against

pesticides concentration. Results were statistically analyzed according to

the method of Litchfield and Wilcoxon (1949). The obtained data

including slopes of the regression lines and LC50 values with thier 95%

confidence limits were recorded.

5-Field studies :

Pestesides used in the laboratory were subjected to further

evaluation at the Experimental Farm of Faculty of Agric., Kafr El-Sheikh,

Tanta Univ. during the growing season, summer 1995. A cultivated area

grown with okra and squash was divided into plots of 1/60 fed. each.

Treatments were distributed in a complete randomized block desigen with

3 replicates. Pesticides used and their application rates are showin in

Table 1.

Table (1): Pesticides used in the field experiment, their formulations

and application rates.

103


Pesticides used Formulation Rate/ fed.Carbosulfan Marshal WP 25% 600 gm.

Pirimicarb Pirimor WP 50% 500 gm.

Propoxur Unden EC 50% 1L.

Chlorpyrifos-methyl Reldan EC 50% 1 L.

Dimethoate Dimethoate 40% 1L.

Phenthoate Cidial EC 50% 0.800 L.

Pirimiphos-methyl-methyl

Actellic EC 50% 1.5 L.

Profenofos Selecron EC 72% 0.750 L.

Pesticides were diluted with water at rate of 200 litter / F. Mature

plants were sprayed with the tested chemicals using a knapsack sprayer

(Model CP3) fitted with one nozzle. Three sprays were applied each

separated by 9 days interval. The first spray was done when insect

infestation was observed on plants. For each crop, three plots were left

untreated as a control check. Inspection of insect infestation was carried

out before spraying and continued 1,3,5, and 7 days after each spray. For

each plot (representing one replicate), the number of larvae of cotton

leafworm, aphids and immature stage (nymphs) & adults of whitefly on 5

leaves (in squash) and 10 (in okra) were recorded. The examined leaves

were randomly collected from each plot.The efficiency of pesticides was

calculated according to the Hinderson and Tilton equation (1955).

%Reduction = 1-a

b

c

d 100

a = Counts in treatments after spraying.

b = Counts in treatments before spraying.

c = Counts in control before spraying at the same interval of 1, 3, 5, 7

days.d = Counts in control after spraying.

The data were statistically analysed using complete randomized block

design.

6- Acute toxicity to rats :

104


The LD50 was assessed according to the method of

Weil (1952), owing to its simplicity and few number of

experimental animals required. For the determination of

LD50, exploratory trials were performed in order to find the

smallest dose of toxic effect to start with. Four dosages

each of 2 animals were tested for each insecticides (n=2).

Rats were given single oral doses each of 1 ml of corn oil

containing the specified amount of the tested insecticide

using special syringe needle equipped with a ball

tip.Mortality rates for rats were recorded after 24 hours

from treatment. The LD50 values and thier confidence

limits, were calculated as follows:

log m = log Da + d(F+1).

as log m = the log of the LD50.

log Da = the log of the lowest dosage level.

d = the log of the constant ratio between dosage levels.

F = Factor obtained from the tables according to the number of animals

dosed per level and number of dose levels used.

In an estimation of a confidence interval that will encompass the

LD50 ( 95 times in 100), we take the value that bounded by anti log (log m

± 2 log m). The following formula was used with the value from the

table where: log m=d. .

7-Daily oral dose treatment :

7.1. Experimental:

105


Six groups of male rats each of seven animals were used for each

insecticide. Additional three groups each of seven rats were reserved as a

control check. For each insecticide, rats were orally given single daily

doses each equivalent to 1/10 or 1/30 LD50. Administration of doses

continued for 30, 60 or 90 days. Rats of control groups were only given

corn oil . All doses were prepared and adminisetred in corn oil using

syringe equipped with needle that has a ball tip. Rats of the last group

were served as control and given corn oil only. Throughout the

experimental period, rats were observed for any mortalities or any

clinical signs. After each specified period rats were weighted, slaughtered

and blood samples were collected. Kidneys and livers were obtained and

kept in formalin 10%.

7.2. Serum preparation :

Blood samples were collected in clean sterile vials, left till clotting

occurred and centrifuged at 5000 r. p. m for 15 min. The obtained sera

were separated and kept frozen till use.

8- Biochemical analysis :

The sera of treated rats were subjected to biochemical analysis that

included, total protien, non-specific esterase, cholinesterase (ChE) ,

glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase

(GOT), alkaline phosphatase, albumin, cholesterol, creatinine, billirubin

and uric acid.

8.1. Determination of total protien :

Determination of total protien was carried out by the method of

Henry (1964) using commercial kits of Diamond Diagnostics, Egypt.

106


This method is based on the fact that protien forms a colored complex

with cupric ions in an alkaline medium.

Reagents :

Reagent(1): protien standard 6 gm/dl.

Reagent(2) (BuretReagent) : sodium hydroxide 0.2 N.

1 sodium, potassium tartarate 18 mmol/l.

1 potassium iodide 6 mmol/l.

1 cupric sulfate 6 mmol/l.

Procedure:

Three tubes, blank, standard and sample tubes were set up. 50 ml of

regeant 1 was added to the standard tube while 50 ml of serum was added

to the sample tube. 2.5 ml of reagent 2 were added to each of the three

tubes. Tubes were mixed well, incubated for 5 min. at 20-25° C and the

produced colour was measured at 546 nm. The absorbance of the sample

(A sample) and the standard (A standard) were read aginst the blank.

Calculation :

Serum protien concentration (g / dl) =A

A6

sample

standard

8.2 Determination of non-specific esterases activity :

Non-specific esterases were determined according to the method of

Van Asperen (1962). The method depends on the enzymatic hydrolysis

of a or -naphthyl acetate producing a or -naphthols. The naphthols

were colorimetrically determined by azo coupling. Because of the

excessive insolubility of naphtholic azo dyes, it was found that , azo dyes

may be kept in a perfectly clear colloidal solution by the use of

detergents. Sodium lauryl sulphate detergent proved to be very efficient.

This detergent had an additional advantage of stopping enzymatic

reaction immediately.

107


Reagents :

1- 310-4 M a-naphthyl acetate containing 1% acetone and 0.04

M phosphate buffer pH 7.

2- Diazoblue - sodium lauryl sulphate solution (2parts of fast

blue B 1% + 5 parts of 5% solution sodium lauryl sulphate)

(DBLS).

Standard cruve for a -naphthol :

Standard curve was obtaind by reacting 6 ml naphthol solution (in

1% acetone and 0.04 M phosphate buffer pH 7 ) of different

concentrations with 1 ml solution of the diazoblue lauryl sulphate

(DBLS). A violet colour immediately develops and quickly changes into

stable colour for a-naphthol which was measured at 600 nm. Blank

contained 6 ml phosphate buffer 0.04 M and 1% acetone and 1 ml DBLS.

a-naphthol concentration was plotted against the obatined optical density

to calculate K value (Fig.1 ).

Measurement of esterase activity :

Five ml of substrate solution (3 10-4 M a-naphthyl acetate

containing 1% acetone and 0.04 M phosphate buffer pH 7 ) was mixed

well with 0.05 ml of serum and the volume was completed to 6 ml with

buffer. After 30 minutes of incubation at 27° C, 1 ml of DBLS was added

and after 5 min., the stable blue colour developed was measured at 600

nm. The specific activity was calculated as m mole a- naphthol /min. /mg

protien.

8.3:Determination of cholinestrase activity:

Cholinesterase of serum was determined according to the method

of Ellman et. al., (1961). This method is based on the hydrolysis of

acetylthiocholine Iodide(ASChI) as substrate by the enzyme ChE to

produce thiocholine and acetic acid. Thiocholine reacts with dithiobis-2-

108


nitrobenzoic acid (DTNB) to produce the yellow color anion of 5-thio-2-

nitrobenzoic acid. The rate of color formation as a function of enzyme

activity is measured spectrophotometrically at 412 nm.

Reagents:

ChE substrate solution:

The solution was prepared by dissolving 10-3M ASChI and 10-3 M

DTNB in a buffer solution containing 100 mM NaCl, 20 mM MgCl2,

109


0 0.05 0.1 0.15 0.2 0.250

0.2

0.4

0.6

0.8

1

Op

tic a

l Den

sity

.

Fig.(1): standard cruve of a -naphthol.

110

20mM Na2HPO4 and the volume was completed with distilled water to

500 ml (pH 7).

Blank solution:

The blank solution was identical to substrate solution except that the

substrate (ASChI) was ommitted.

Procedure:

1- 4 ml of substrate solution were added to two tubes and 4 ml of

the blank solution to a third tube.

2- Tubes were preheated to 37°C in water bath and 50 µl of

serum sample were added.

3- O.D readings were taken at two time points 1 minute a part at

412 nm.. Thus, changes of optical density per minute

(O.D/min.) were obtained.

Calculation:

The activity in µ moles ASCh/min/mg protein was calculated by using the following equation:

Activity = ( O.D Average sample - O.D blank) fv 10

pc sv t

6 32258

Where : fv = final volume (0.004 L)

pc = protein concentration (mg/ml).

sv = sample volume

t = time of incubation (1 minute).

8.4.Determination of transaminases:

The colorimetric method of Reitman and Frankel (1957) was used for the

determination of Aspartate aminotransferase (formerly Glutamate

oxaloacetate transaminase GOT) and Alanine aminotranasferase (formerly,

Glutamate pyruvate transaminase, GPT) activity. Commercial kits of

111

bioMérieux, France were used. The principle of determination was based

on the following reactions:

GOT:Asparate+a-keto-glutarate GOT oxaloacetate+ glutamate

GPT: Alanine+a-keto-glutarateGPT

pyruvate+glutamate

The pyruvate or oxaloacetate formed is measured in its derivative form,

2,4- dinitrophenylhydrazone.

Reagents:

Reagent 1(GOT substrate): Phosphate buffer pH 7.5 85 mol/L

aspartate 200 m.mol/L

a- ketoglutarate 2.0 m.mol/L

Reagent 2 (GPT)

substrate):

Phosphate buffer pH 7.5 95 m.mol/L

alanine 200 m.mol/L

a-keto glutarate 2.0 m.mol/L.

Reagent 3 (Color reagent): 2,4 dinitro phenyl hydrazine 1 m.mol/L.

Reagent 4 (Standard): Pyruvate

Procedure:

a- Standard curve:

Different aliquots of distilled water, reagent 1; reagent 4 and

reagent 3 were pipetted into each of six test tubes as shown in the

following table:

Tube No. 1 2 3 4 5 6

Distilled water: 0.2 0.2 0.2 0.2 0.2 0.2

Reagent 1 1.0 0.9 0.8 0.7 0.6 0.5

Reagent 4 -- 0.1 0.2 0.3 0.4 0.5

Reagent 3 1.0 1.0 1.0 1.0 1.0 1.0

The mixture of each tube was shaken well and let stand for 20

minutes at room temperature, then 10 ml of NaOH 0.4 N was added to

each tube and the contents of each tube were mixed well and let to stand

112

for 5 minutes. The developed color was measured at 505 nm. The

standard curve was drawn by plotting the number of units/ml on the

abscissa and the optical denisty on the ordinate.

Tube No.: 1 2 3 4 5 6GOT units/ml: 0.0 22 55 95 150 215

GPT units/ml : 0.0 25 50 83 126 ----

Measurment:Two test tubes were set up for serum samples as following:

1- One ml of reagent 1 (GOT substrate) was pipetted into one tube and one

ml of reagent 2 (GPT substrate) was pipetted in the second tube, then the

tubes were incubated for 5 minutes at 37°C.

2- 0.2 ml of serum was added to each tube and the contents were mixed well

and incubated at 37°C for exactly one hour (GOT tube) and 30 minutes

(GPT tube), then 1 ml of reagent 3 was added to each tube. The contents of

each tube were let to stand for 20 minutes at room tepmerature.

3- After that, 10 ml NaOH 0.4 N were added to each tube and the contents

were mixed well.

4- After 5 minutes the devloped colour was measured under the same

conditions used for the standard curve.

Calculation:The number of GOT and GPT units/ml of serum was calculated from

the standard curve as shown in figure 2 and 3.

8.5.Dtermination of alkaline phosphatase:

The method of Bessey et al (1946) were used for the determination of

alkaline phosphatase. The determination was done using commercial kits of

113

0 50 100 150 200

(Units/ml.)

0

0.1

0.2

0.3

0.4

0.5

Opt

i cal

De n

sity

.

Fig. ( 2): Standard cruve of GOT.

114

0 50 100 150

(Units/ml.)

0

0.2

0.4

0.6

0.8

1

Op

tica

l Den

s ity

.

Fig ( 3): standard cruve of GPT.

115

Biocon Diognostik, Germany . This method is based on the following

reaction:

P-nitrophenyl phosphate+H2O AP Phosphate +P-nitrophenol

Reagents:

Reagent 1 : (Buffer). glycine buffer pH 10.5 50 mmol/l

MgCl2 0.5 mmol/l.

Reagent 3:(Substrate solution): p- nitrophenyl phosphate 5.5 mmol/l.

Reagent 4 : (Standard ): 4-nitro phenol 900 umol/l.

Additional reagent solution : Sodium hydroxide 0.02 N.

Procedure:1- Preparing working solution:

10 volumes buffer (R1) + 1 volume substrate solution (R3) . This working

solution is stable for one week at +2 to +8° C .

2- Two tubes were prapared for blank and sample. 500ml working solution

were added to each tube. 50ml serum were added to sample tube only. Tubes

were mixed well, incubated for exactly 30min. in water bath at +37 ° C.

3- After that 5 ml of sodium hydroxide solution were added to tubes of

sample and blank.50 ml serum were added to blank tube. Tubes were mixied

well and poured into cuvettes. Absorbance (A) of sample at 405 nm was

recorded.

Calculation:Akaline phosphatase activity (U/l) = A405nm 200

8.6.Determination of cholesterol:

Commercial kits of Biocon Diagnostik, Germany were used. The

enzymatic colorimetric method of Flegg (1973) as modified by Richmond

(1973) and Thomas et al. (1992) was used for cholesterol determination.

The determination is based on the principle that cholesterol and its esters are

released from lipoproteins by detergents. Cholesterol esterase hydrolizes the

esters and H2O2 is formed in the subsequent enzymatic oxidation of

cholesterol by cholesterol-oxidase according to the following reaction:

116

Cholesterol ester + H2O Cholesterol Esterase

Cholesterol + fatty acids

Cholesterol + O2Cholesterol Oxidase

Cholesten-3-on + H2O2/2 H2O2 +

Phenol + 4-aminophenazone Peroxidase quinoneimine dye + 4H2O

Reagents:

Reagent(1):(Buffer): pipes buffer, pH 6.9 90mmol/l.

1 phenol 26 mmol/l.

Reagent(2): (Enzyme reagent): cholesterol oxidase 200 U/l.

cholesterol esterase 300 U/l.

4-amino phenazone 0.4 mmol/l.

Reagent (4): (Standard): cholesterol 200 mg/dl

Procedure : 1-Contents of enzyme reagent (R2) were dissolved with the

corresponding volume of buffer (R1). This working solution was stable 2

weeks at +20 to

+25 °C or 8 weeks at +2 to +8 °C.

2- Three tubes, sample tube, standard tube and blank tube were prepared.

10 ml of reagent 4 (standard) were added in stanadard tube. 10 ml of

serum were added in sample tube. 1000 ml working reagent were added to

all tubes. Tubes were mixed well, incubated at: 37 °C for 5 minutes and

the developed colour was measured spectrophotometrically at 546 nm.

Calculation:

Cholesterol in mg / dl = sample

standard standard concentration

Standared concentration: 200 mg/dL.

8.7.Determination of total bilirubin:

Commercial kits of Diamond Diagnostics, Egypt, were used.The

determination was carried out according to the method described by

Jendrassik et al. (1938). The determination is based on the idea that

117

bilirubin is coupled with diazotized sulfanilic acid in the presence of

caffeine to gave an azo dye.

Reagents:

Reagent(1): Sulfanillic acid 31 mmol/l

1 HCl 0.2 N

reagent(2): Sodium nitrite 28 mmol/l.

Reagent(3): Caffeine 0.28 mol/l.

1 sodium benzoate 0.55 mol/l.

Reagent(4): Tartarate 0.99 mol/l.

1 NaoH 2.0N.

Procedure:1- Two clean and dry tubes for sample blank and sample were prepared.

0.2 ml of reagent 1 was added to sample blank and sample, 1 drop of

reagent 2 was added to sample tube, 1 ml of reagent 3 were added to each

of sample blank and sample tubes.

2- 0.2 ml of serum was added to each of sample blank and sample tubes.

Tubes were mixed and incubated for 10 min. at 20-25 °C. after that 1 ml

of reagent 4 was added to each of tubes. Tubes were mexid and incubated

for 5 min. at 20-24 °C.

3- Absorbance of sample against sample blank (A sample) was read at

the wave lenghth 578 nm.

Calculation:

Total bilirubin (mg/dl)= (A sample) 10.8

8.8.Detemination of Albumin :

The colorimetric method of Doumas and Watson (1971) as

modified by Webster (1974) was used for assaying Albumin using

bromcresol green (BCG) at pH 4.2. Commercial kits of Biocon

Diagnostik, Germany were used.

Reagents :

Reagent(1):(BCG reagent): succinate buffer, pH 4.2 75 mmol/l.

118

1 Bromcresol green 0.15 mmol/l.

1 Brij 35 7ml/l.

Reagent(4):(Standard): bovine albumin 5g/100 ml.

Procedure :

Three tubes, blank tube, standard tube and sample tube were set up

and treated as follows:

1- 20ml of serum was transferred into sample tube. 20ml of reagent 4

(standard) was transferred into standard tube.

2- 4000 ml of reagent 1 was added to each tube of sample, standard and

blank, mixed well and measured after 10 minutes. Results against reagent

blank were read at 628 nm.

Calculation:

Albumin in g / dl = A sample

A standardstandard concentration

Concentration of standard: 5gm/100 ml.

8.9.Determination of creatinine :

The determination was carried out according to the method of Henry

(1974).Commercial kits of Diamond Diagnostics, Egypt, were used.The

detemination is based on the idea that creatinine in alkaline solution

reacts with picrate to form a colored complex.

Reagents:

Reagent(1): Creatinine standard 2mg/dl.

Reagent(2): color reagent: picric acid 38 mmol/l.

Reagent(3):alkaline reagent: sodium hydroxide 1.2 mmol/l.

Reagent(4): equal volumes of solution 2and 3 were mixed.

119

Additional reagent: Trichloro acetic acid (TCA) 1.2 mol/l.

Procedure:

1- The serum was firstly deprotenized by pipetting 1 ml of trichloroacetic

acid and 1 ml of serum into centrifuge tube. The mixture was mixed well

and centrifuged for 10 min. at 3000 r.p.m, then the clear supernatant was

porured into dry test tube.

2- Three clean and dry tubes for blank, standard and serum supernatant

were prepared. 0.5 ml of distilled water was added to the blank tube and

0.5 ml of reagent 1 was added to the standard tube. 0.5 ml of

trichloroacetic acid was added to each of blank and standard tubes while 1

ml of serum supernantant was added to serum tube.

3- For all tubes, 1 ml of reagent 4 was added to each, mixed and

incubated for 10 min. at 37° C. The absorbances of sample (A sample)

and standard (A standard) were measured against the blank at 520 nm.

Calculation :

Creatinine (mg / dl) =A sample

A standard2

8.10.Determination of uric acid:

The enzymatic colorimetric test of Pileggi and Barthelmai (1962)

as modified by Trinder (1964) was used for determination of uric

acid.Commercial kits of Biocon Diagnostik, Germany were used. The

determination is based on Trinder reaction:

Uric Acid + O2 + H2O Uricase Allantion + CO2 + H2O2

2H2O2 + 4 Aminophenazone + 2-4 Dichloro-phenol-Sulfonate

Quinoneimini Dye + 4H2O

Reagents :

Reagent(1):(buffer): hepes buffer pH 7.4 50mmol/l.

120

1 2,4 HDCBS 4 mmol/l.

Reagent(2):(Enzyme reagent):uricase 60 U/l.

peroxidase 660 U/l.

1 4-Aminophenazone 1mm ml/l.

Reagent(4):(standard): uric acid 6 mg/dl

Procedure :

1-Contents of enzyme reagent (2) were dissolved with the corresponding

volume of buffer(R1). This working solution was stable 2 weeks at +20 to

+25 °C

2- Three tubes were prapared for blank, standerd and sample. 1000 ml of

working solution were added to each tube. 20 ml serum were added to

sample tube only, 20 ml of reagent 4 were added to standard tube. Tubes

were mixed well, incubated for exactly 5 min. in water bath at +37 °C.

Absorbance (A) of sample against blank at 546 nm.was read.

Calculation:

Uric acid in mg / dl = sample

standardstandard concentration.

Standard concentration: 6mg/dl.

9. Histopathological studies:

This experiment was conducted to study the histopathological

lesions of liver and kidney tissues of rats treated with 1/10 and 1/30 LD5 0

of chloropyrifos methyl or pirimicarb as daily oral dose for 30, 60 or 90

days. The livers and kidenys of slaughtered rats were removed and

prepared for histopathological examination according to Lillie and

Fullman (1976) .

Chemicals and apparatus:

1-Alcohol of different concentrations (i.e.. 70%, 80%, 90%, 96%

and100%)

121

2-Formalin 10%, 3-Xylol, 4-Paraffin wax(melting point of 50-56 °C),

5- Mercuric oxide 6- acetic acid 7- Canda balsam, 8-Water bath, 9-

Glass slides, 10-Cover glass, 11-Microtome (Model MR52, Russian).

12-Microscope. 13-Incubator.

Haematoxylin stain

The components of Harris alum hematoxylin-stain were prepared

as follows:

A- Haematoxylin crystal 5 gm

Absolute alcohol 50 ml

B- Potassium alum 100 gm

Distilled water 1000 ml

The solution B was boiled, and added to solution A, then 2.5 gm of

mercuric oxide were added, stirred , boiled for 1-2 min.and cooled

quickly by immersing the container into cold water. The mixture was

allowed to stand overnight and acidiffied by adding 30-50 ml acetic acid

to inhibit staining of cytoplasm. Finally it was filtered and stored in a

tightly stoppered bottle.

Eosin 1%:

1- Preparation of stock solution:

Eosin 1 gm

Alcohol (80%) 100 ml.

2- Working Eosin solution

Eosin 1% stock solution 1 part.

Alcohol 80% 3 part

Procedure:

1- Fixing , hardening and dehydration:

Specimens, from kidneys, and livers of rats were excised, collected

and fixed in 10% neutral buffered formalin.The tissues were passed

122

through ascending grades of alcohol (70%, 80%, 90%, 96% and 100%).

The dehydration was not done rapidly in order to avoid distoration and to

add more hardness to the tissues. Tissues were mascerated in xylol.

2- Embedding in paraffin wax and sectioning:

Tissues were mascerated in xylol/ paraffin wax mixture (1:3). After

complete infiltration, a solid rectangular block of paraffin was made.

Microtome was used for cutting blocks into sections each of thickness 5

mm.

3- Mounting paraffin sections and staining:

The ribbons were mounted on clean glass slides pretreated with thin

film of Mayer`s albumin fixative. The slides were put on a hot plate (50

°C) over nigh to dry. The sections were then stained with differentially

double stain, Ehrlish haematoxylin for nuclei and Eosin for cytoplasm and

cell wall. The staining process was carried out as follows:

The slides were dipped in xylol for 20-30 minutes to remove the

wax from the sections, then dipped again in fresh xylol for 15 minutes to

ensure complete removal of wax. The slides were then dipped in a series

of different alcohol solutions each for 5 minutes starting with absolute

ethanol, 95, 90, 80, and 70% ethanol. Then the slides were rinsed in

distilled water and became ready for staining. They were dipped in the

solution of Ehrilish stain for 2 minutes. The excess stain was washed in

distilled water, 70% acidic alcohol until the sections became reddish, then

treated with alkaline alcohol 70% until the sections turned blue. Then

slides were dipped in 80, 90 and 95% alcohol each for 3 minutes,

respectively, then in Eosin for a few seconds. Excess Eosin was removed

with 95% alcohol. To remove any traces of water, the slides were dipped

in absolute ethanol for 5 minutes then in a series of xylol until perfectly

clear sections were obtained. The sections were covered with Canada

balsam and glass covers were placed properly on the slides. Finally

123

sections were examined under the microscope. Nuclei appeared blue,

while cytoplasm had the red color.

10-Statistical analysis:-

Statistical analysis of all data was carried out according to: Duncan`s

multiple range test (Duncan, 1955.).

124

RESULTSRESULTS ANDAND

DISCUSSIONDISCUSSION

RESULTS AND DISCUSSION

1-Pesticidal activity of tested chemicals:

1.1 Laboratory tests:

The insecticides were evaluated for their toxicity against the cotton

leafworm, Spodoptera littoralis; three species of aphids (i.e. Aphis

gossypii, Aphis craccivora and Brevicoryne brassicae), the spider mite,

Tetranychus cinnabarinus and the predator Paederus alfierii.

1.1.1 Toxicity to the cotton leafworm, S. littoralis :-

The leaf dipping technique was adopted to evaluate the toxicity

of the insecticides to the fourth instar larvae. Results are recorded in

Table (2) and depicted in Fig. (4). According to LC50 values,

chloropyrifos-methyl exhibited the highest toxicity to S. littoralis

followed by profenofos (LC50`s: 74.53 and 133.35 ppm, respectively).

Weak toxicities was exhibited by pirimiphos-methyl, phenthoate and

carbosulfan (LC50`s: 1485.7, 3949.3 and 4855.4 ppm, respectively). As

for pirimicarb, propoxur and dimethoate, they were nearly non-toxic to S.

littoralis (LC50`s:>10000 ppm).The results are somewhat in agreement

with those of other authors. El- Dahan (1983) evaluated five synthetic

pyrethroids, three OP compounds and one carbamate insecticide for their

toxicity against the fourth instar larvae of S. littoralis. The OP compound,

profenofos was found to be the most toxic one. Chloropyrifos-methyl and

profenofos were found also to be effective against the same insect in

other work (Ghattas and El-Keie, 1986; Ascher and Nemmy, 1990 and

El- Dahan, 1991). Of four insecticides tested against S. littoralis,

dimethoate was the least effective one (Fahmy, et al. 1979). El-Gayar et

al. (1979) found that, pirimiphos-methyl was classified among other

insecticides as less potent against 2 nd and 4 th instar larvae of S. littorallis.


1.1.2. Toxicity to aphids:-

Laboratory tests were conducted to evaluate the toxicity of the

tested insecticides to the aphids, Aphis gossypii, Aphis craccivora and

Brevicorne brassicae. The slide dipping technique was applied. Results

are recorded in Tables (3, 4, 5) and probit regression lines are shown in

Fig. (5, 6, 7). These results revealed that, the toxicity to aphids greatly

varied depending on the type of insecticide and/or the species of the

tested aphid. Against A. gossypii, profenofos had the highest toxicity

followed by dimethoate, phenthoate, chloropyrifos-methyl, carbosulfan

and pirimicarb (LC50`s values: 0.004, 0.01, 0.05, 0.11, 0.125 and 0.55

ppm, respectively). Propoxur and pirimiphos-methyl showed very weak

toxicity (LC50`s: 26.4 and 66.78 ppm, respectively). For the aphid, B.

brassicae, chloropyrifos-methyl was the most toxic compound followed

by profenofos, pirimicarb, propoxur and carbosulfan (LC50`s: 1.44, 2.40,

2.95, 4.37 and 5.88 ppm, respectively). Dimethoate, phenthoate and

pirimiphos-methyl were of weak toxicity (LC50`s: 32.46, 18.5 and 8.64,

respectively). The aphid, A. cracivora was the least susceptible to the

tested insecticides. The most toxic compound was profenofos followed by

pirimiphos-methyl and dimethoate (LC50`s: 5.45 and 8.25, 11.56 ppm,

respectively). Phenthoate showed very weak toxicity (LC50: 1632.42

ppm). When the toxicity against the three species of aphids is taken in

consideration, profenofos, dimethoate and chloropyrifos-methyl were

considered to be the most toxic compounds among the tested insecticides.

The results partly agreed with those of El-Sebae and Saleh, (1970);

Singh and Sircar, 1983; Salama et al, 1984; Thakur et al. 1984;

Sandhu, 1986; Zheng and Wing, 1987; Zidan 1989b; Gao et al.

126


Table (2): Toxicity of the tested pesticides against Spodoptera littoralis using leaf

dipping technique.

Pesticides LC50 (ppm.) Confidence limits. Slope.

Lower UpperCarbosulfan. 4855.42 3564.54 6613.79 2.66

Pirimicarb. >10000

Propoxur. >10000

Chlorpyrifos-methyl. 74.53 62.58 88.77 3.33

Dimethoate. >10000

Phenthoate. 3949.29 3175.22 4912.07 5.34

Pirimiphos-methyl. 1485.68 1191.04 1853.21 5.27

Profenofos. 133.35 114.61 155.15 4.44

Table (3): Toxicity of the tested pesticides against Aphis gossypii using slide dipping

technique.

Pesticides LC50 (ppm.) Confidence limits Slope.


Pirimicarb. 0.55 0.32 0.93 0.96

Propoxur. 26.40 18.02 38.68 1.32

Chloropyrifos-methyl. 0.11 0.074 0.16 1.56

Dimethoate. 0.01 0.0037 0.03 0.54

Phenthoate. 0.051 0.024 0.11 0.57


Profenofos. 0.0044 0.0037 0.0051 3.76

Table (4): Toxicity of the tested pesticides against Brevicoryne brassicae using slide

dipping technique.

127




Pirimicarb. 2.95 1.70 5.12 1.83

Propoxur. 4.37 2.93 6.52 1.79


Dimethoate. 32.46 20.34 51.80 2.16

Phenthoate. 18.50 10.58 32.35 1.28


Profenofos. 2.40 1.75 3.29 3.20

Table (5): Toxicity of the tested pesticides against Aphis craccivora using slide

dipping technique.



Pirimicarb. 17.50 12.68 24.14 3.13

Propoxur. 153.90 101.89 232.46 1.73


Dimethoate. 11.56 7.50 17.82 2.33

Phenthoate. 1632.42 818.10 3257.28 1.03


Profenofos. 5.45 4.34 6.85 3.13

128


10 100 1000 10000 100000

Concentration (ppm).

0

50

100

% M

o rta

l ity.

2

3

4

5

6

7

8

Probit.

carbosulfan

pirimiphos-methyl

chlorpyrifos-methyl

phenthoate

profenofos

Fig. (4) Probit regression lines for the toxicities of various pesticides to Spodoptera littoralis.

129


0.001 0.01 0.1 1 10 100 1000


0

50

100

% M

o rta

l ity.

2

3

4

5

6

7

8

Probit.

carbosulfan

pirimicarb

propoxur

pirimiphos-methyl

chlorpyrifos-methyl

phenthoate

profenofos

dimethoate

Fig. (5) Probit regression lines for the toxicities of various pesticides to Aphis gossypii .

130


0.1 1 10 100 1000


0

50

100

% M

o rta

l ity.

2

3

4

5

6

7

8

Probit.

carbosulfan

pirimicarb

propoxur

pirimiphos-methyl

chlorpyrifos-methyl

phenthoate

profenofos

dimethoate

Fig. (6) Probit regression lines for the toxicities of various pesticides to Brevicoryne brassicae.

131


0.1 1 10 100 1000 10000


0

50

100

% M

o rta

l ity.

2

3

4

5

6

7

8

Probit.

carbosulfan

pirimicarb

propoxur

pirimiphos-methyl

chlorpyrifos-methyl

phenthoate

profenofos

dimethoate

Fig. (7) Probit regression lines for the toxicities of various pesticides to Aphis craccivora.

132


1990; Zeitoun et al. 1990; Sharma et al, 1991; Mourad, 1991 & Konar

and Rai, 1992 who showed that dimethoate was effective against aphids.

Against A. gossypii, profenofos proved to be the potent among the tested

chemicals (Zeitoun et al 1990). Dimetry and Marei, (1992). Also they

found that, profenofos was highly toxic to the cabbage aphid, B.

brassicae.Rizk and Kamel (1991) found that, chloropyrifos-methyl

proved to have an immediate effect on aphids. For pirimicarb, Zidan et

al, 1988a and 1989b reported that the compound have shown to be very

effective against A. gossyppii and Schizaphi graminum aphids. However

Abdel-Wahab and Mohamed, (1992) found that profenfos was the most

potent compound against S. graminum aphid whereas pirimicarb was the

least toxic one.

Results of the present investigation show that A. gossypii was

the most susceptible species to carbosulfan. Similar results were obtained

by Shalaby et al. 1991; Halawa et al. 1992; Mourad, 1991; Shaheen et

al 1992 and Nassef, et al. 1995 who found that, carbosulfan was among

the most effective pesticides against A. gossypii.

1.1.3 Toxicity to the spider mite, T. cinnabarinus:

The leaf disc dipping technique was used to evaluate the toxicity of

the tested insecticides to adults of mites Teteranychus cinnabarinus.

Results are recorded in Table (6) and probit regression lines are shown in

Fig(8). Results show that, profenofos was the most toxic compound

followed by pirimiphos-methyl and phenthoate (LC50`s: 186.54, 223.16

and 688.15 ppm, respectively). The rest of the tested insecticides showed

very weak toxicity against the spider mite. Thomson, (1983) reported

that, profenofos and phenthoate had acaricidal properties.

In general, the very weak toxicity exhibited by the tested

insecticides is something logical since these toxicants are not specific

acaricides.

133


1.1.4. Toxicity to the predator, Paederus alfierii:

Surface deposit technique was used to determine the toxicity of the

insecticides to the adults of predator, P. alfierii. LC50 values and their

confidence limits are recorded in Table (7) and probit regression lines are

shown in Fig.(9). The toxicity of the compounds could be arranged

descendingly as follows: Propoxur> phenthoate> dimethoate>

carbosulfan> pirimicarb> pirimiphos-methyl> chloropyrifos-methyl>

profenofos (LC50`s: 0.0027, 0.0029, 0.0039, 0.0062, 0.1015, 0.1148,

0.5485 and 0.8113 mg/cm2, respectively. Based on the obtained data, the

insecticides could be classified into two groups according to their

toxicities to the predator P. alfierii: The first group includes the

insecticides that exhibited high toxicity (i.e. propoxur, phenthoate,

dimethoate and carbosulfan). The second group includes the insecticides

that showed moderate to low toxicity (i.e. pirimicarb, pirimiphos-methyl,

chloropyrifos-methyl and profenofos). Similar results were obtained by

Zeitoun et al. (1990) and Hussein et al. (1993a) who found that

profenofos (among other insecticides) had the least toxic effect against

the Coccinellid predator, Coccinella undecimpunctata when applied in

the field. Against the aphidivorous coccinellid Coccinella

septempunctata, chlorpyrifos-methyl was among the most safe

compounds when applied topically under laboratory conditions (Hussein

et al. 1981). In contrast to the results of the present investigation,

carbosulfan was much safer on natural enemies (including P. alfierii)

under field conditions (Darwish and Farghal 1990).

Table (6):Toxicity of the tested pesticides against Tetranychus cinnabarinus exposed to

leaf disk dipping technique.

134


Pesticides LC50 (ppm.) Confidence limits Slope.


Pirimicarb. 16471.34 11795.8 23000 1.51

Propoxur. 36065.23 23137.9 56215.1 1.31


Dimethoate. 1079.19 830.96 1401.59 2.72

Phenthoate. 688.15 545.86 867.55 2.51


Profenofos. 186.54 124.96 278.46 1.78

Table (7): Toxicity of the tested pesticides against Paederus alfierii using surface

deposit technique.

Pesticides LC50 (mg/cm2) Confidence limits Slope.


Pirimicarb. 0.1015 0.08223 0.1253 3.90

Propoxur. 0.0027 0.00194 0.0039 2.30


Dimethoate. 0.0039 0.00305 0.0050 3.23

Phenthoate. 0.0029 0.00161 0.0055 0.84


Profenofos. 0.8113 0.59927 1.0985 2.71

135


10 100 1000 10000 100000


0

50

100

% M

o rta

l ity.

2

3

4

5

6

7

8

Probit.

Carbosulfan

Pirimicarb

Propoxur

Pirimiphos-methyl

Chlorpyrifos-methyl

phenthoate

Profenfos

dimethoate

Fig. (8) Probit regression lines for the toxicities of various pesticides to Tetranychus cinnabarinus .

136


0.0001 0.001 0.01 0.1 1 10

Concentration

0

50

100

% M

o rta

l ity.

2

3

4

5

6

7

8

Probit.

carbosulfan

pirimicarb

propoxur

pirimiphos-methyl

chlorpyrifos-methyl

phenthoate

profenofos

dimethoate

(mg/cm 2).

Fig. (9) Probit regression lines for the toxicities of various pesticides to Paederus alfierii.

137


Larvae of the three predators, Chrysopa carnea, Coccinella

undecimpunctata and Paederus alfierii were highly susceptible to

dimethoate (Shaheen and Kaddy, 1987). Generally, it is well known

that, the wide and unwise application of different pesticides leads to

harmful effects on the beneficial insects especially entomophagous. One

of the solutions of this problem is the use of selective pesticides as a

possible method of integrated control. So, the studies connected with the

impact of different pesticides on predators and parasites can facilitate the

determination of selectivity of these pesticides. Such studies are therefore

helpful for planning more effective IPM programs.

Data of toxicity of the tested insecticides (calculated as LC50`s and

toxicity indexes) against different pests and the tested predator are

summarized in Table (8).

1.2. Field studies:

The insecticides were further evaluated under field conditions.

Three sprays (each separated by 9 days intervals) were applied on squash

and okra plants. Plants were examined before and after each spray for

their infestation with cotton leafworm, aphids, mites and whitefly.

Percents of pest reduction were determined using Hinderson and Tilton

equation (1955). Aphids, A. gossypii and whitefly, B. tabaci were mainly

observed and detected on squash plants while the white fly and the cotton

leafworm S. littoralis could be observed on okra plants.

1.2.1 The cotton leafworm, S. littoralis :

The cotton leafworm could be observed only on okra plants.

Results are recorded in Tables 9,10 and depicted in Fig 10.

These results revealed that, profenofos and chloropyrifos-methyl

had the highest effects against the cotton leafworm (Average of %

reduction throughout the experiment period were 83.52 and 81.5 ,

138


Table(8): Toxicity of the tested pesticides against certain pests and the beneficial predator, P. alfierii.

Insects S.littorals A. gossypii. A. craccivora. B.brassica. T.cinnabarinus. P. alfierii

Pesticides LC50 Toxicity LC50 Toxicity LC50 Toxicity LC50 Toxicity LC50 Toxicity LC50 Toxicity

(ppm) index. (ppm) index. (ppm) index. (ppm) index. (ppm) index. (mg/cm2) index.Carbosulfan. 4855.42 1.53 0.125 3.52 34.22 15.92 5.88 24.48 5494.52 3.3950 0.00626 44.40

Pirimicarb. >10000 0.55 0.8 17.50 31.14 2.95 48.81 16471.3 1.1325 0.10153 2.738

Propoxur. >10000 26.40 0.016 153.90 3.54 4.37 32.95 36065.2 0.51722 0.00278 100

Chloropyrifos-methyl. 74.53 100 0.11 4 8.83 61.72 1.44 100 2525.23 7.3870 0.54858 0.5027

Dimethoate. >10000 0.01 44 11.56 47.14 32.46 4.43 1079.19 17.2851 0.00393 70.73

Phenthoate. 3949.29 1.887 0.051 44.56 1632.42 0.33 15.50 9.29 688.15 27.10 0.00298 93.28

Pirimiphos-methyl. 1485.68 5.01 66.78 0.006 8.25 66.06 8.64 16.66 223.16 83.59 0.11482 2.421

Profenofos. 133.35 55.89 0.0044 100 5.45 100 2.40 60 186.54 100 0.81139 0.342

Toxicity index calculated according to Sun equation, 1950.

Toxicity index =LC50 of the most toxic one/ LC50 of the insecticides 100.

Table ( 9): Efficiency of certain pesticides against the population density of cotton leaf worm, Spodoptera littoralis larvae in okra plants. 139


Chemical Average no. of larvae/3leaves ± S.D at days:-used Before

1st

spray.1 3 5 7 9* 10 12 14 16 18** 19 21 23 25

Carbosulfan 12.25±

0.20

1.25±

0.11

4.25±

0.11

3.11±

0.11

0.28±

0.01

4.42±

0.38

1.43±

0.18

4.35±

0.15

6.10±

0.19

5.42±

0.19

7.33±

0.21

0.67±

0.07

5.05±

0.65

4.36±

0.14

3.54±

0.04

Pirimicarb 2.70±

0.26

1.70±

0.31

1.36±

0.03

0.70±

0.02

0.08±

0.01

3.83±

0.20

4.35±

0.30

4.30±

0.17

5.51±

0.08

4.25±

0.19

5.54±

0.33

5.45±

0.12

4.96±

0.58

3.61±

0.85

2.63±

0.14

Propoxur 4.47±

0.36

1.42±

0.06

1.74±

0.06

1.16±

0.03

0.10±

0.01

2.31±

0.23

2.42±

0.28

2.68±

0.26

3.48±

0.15

2.93±

0.21

4.39±

0.33

2.75±

0.48

3.54±

0.86

2.45±

0.64

2.98±

0.31

Chlorpyrifos-methyl

2.56±

0.23

0.01±

0.001

0.02±

0.002

0.11±

0.01

0.02±

0.002

4.29±

0.29

0.08±

0.001

0.19±

0.01

1.40±

0.17

1.92±

0.06

4.52±

0.27

0.17±

0.15

0.15±

0.08

0.35±

0.10

0.64±

0.10

Dimethoate 9.02±

0.21

5.97±

0.45

3.92±

0.16

2.50±

0.16

0.31±

0.02

3.74±

0.29

4.33±

0.10

4.52±

0.17

5.21±

0.34

4.93±

0.28

5.70±

0.39

5.86±

0.91

3.89±

0.39

4.52±

1.13

2.92±

0.47

Phenthoate 10.23±

0.36

1.19±

0.08

1.52±

0.25

1.43±

0.18

0.18±

0.01

2.67±

0.21

0.80±

0.02

1.03±

0.04

2.43±

0.14

0.96±

0.06

2.33±

0.13

0.45±

0.14

0.60±

0.26

0.85±

0.13

0.85±

0.12

Pirimiphos-methyl

2.63±

0.24

0.25 ±

0.02

0.51 ±

0.02

0.41 ±

0.02

0.05 ±

0.01

2.34 ±

0.27

0.34 ±

0.05

1.28 ±

0.26

2.03 ±

0.04

2.36 ±

0.19

4.79 ±

0.39

1.25 ±

0.19

1.80 ±

0.30

1.45 ±

0.22

2.21 ±

0.24

Profenofos 16.85±

0.46

0.05±

0.002

0.02±

0.003

0.12±

0.003

0.10±

0.13

3.44±

0.17

0.06±

0.01

0.49±

0.06

0.99±

0.03

0.95±

0.10

4.39±

0.30

0.24±

0.21

0.06±

0.01

0.04±

0.01

0.15±

0.03

Control 9.86±

0.68

4.66±

0.38

3.61±

0.17

2.56±

0.29

0.23±

0.02

3.59±

0.35

4.24±

0.13

4.45±

0.20

5.41±

0.13

4.72±

0.10

5.65±

0.24

6.10±

0.73

4.63±

0.34

3.69±

0.92

2.83±

0.36

* the second spray (population density just before spraying) ,** the third spray(population density just before spraying) . S.D: Standard deviation.

140


Table (10): Efficiency of certain pesticides against larvae of cotton leafworm, Spodoptera littoralis in okra plants .

Chemical Rate/ Average % reduction ± S.D. at days:- Averageused fed. 1 3 5 7 9* 10 12 14 16 18** 19 21 23 25 reduction

Carbosulfan 600 gm. 78.34±

2.17 d

5.13±

0.91 a

2.19±

2.56 a

0.35±

0.24 a0.00 a

72.17±

5.51 b

19.95±

8.24 b

7.845±

10.13 a

6.21±

7.06 a0.00 a

81.59±

6.95c

15.73±

0.57c

7.28±

1.88a

1.42±

0.60a

22.66

Pirimicarb 500 gm.0.00a 0.00a

0.14±

0.01 a0.00 a 0.00 a

3.83±

0.26 a

9.42±

0.38 a0.00 a

2.29±

0.14 a

8.09±

0.35 a

8.88±

0.13 a0.00 a

0.22±

0.01 a

5.22±

0.45 a

2.72

Propoxur 1L. 32.78±

2.36b0.00 a

0.05±

0.01 a

4.09±

0.950.00 a

11.29±

0.59 a

6.40±

0.36 a0.00 a

3.52±

0.47 a0.00 a

41.97±

2.35b

1.59±

0.01 a

14.54±

0.16 a0.00 a

8.30

Chlorpyrifos-methyl

1 L. 98.88±

0.32 f

97.52±

0.47 d

82.65±

2.01 c

62.79±

7.38 b

51.07±

4.52c

98.33±

0.15 d

96.4±

0.43 c

86.95±

1.39c

72.7±

1.099c

38.65±

7.76c

90.72±

0.25d

95.11±

2.44f

84.57±

2.35+

75.96±

3.08c

81.52

Dimethoate 1L.0.00a 0.00 a 0.00 a 0.00a 0.00 a

1.97±

0.01 a

2.50±

0.19 a0.00 a 0.00 a

3.16±

0.23 a

4.77±

0.14 a

16.71±

0.28b0.00 a 0.00 a

2.07

Phenthoate 0.800 L. 75.29±

2.39 c

59.25±

7.55 c

45.97±

8.43 b

24.60±

1.58 a

21.31±

3.38b

77.90±

1.53b

63.35±

0.94c

49.61±

2.48b

34.02±

4.71b

19.16±

1.65b

76.58±

1.20c

61.27±

1.30d

52.55±

3.72b

37.63±

2.52b

49.89

Pirimiphos-methyl

1.5 L. 80.11±

1.70d

47.15±

3.978 b

40.08±

7.01 b

11.63±

11.10 b0.00 a

87.40±

2.56 c

55.53±

9.85 c

41.69±

8.13 b

21.99±

14.91 ab0.00 a

75.86±

1.81 c

53.71±

11.16 d

53.12±

11.10 b

7.55±

13.24 a

41.13

Profenofos 0.750 L. 99.33±

0.046 f

99.63±

0.03 d

97.36±

0.11 d

93.63±

0.15e

43.88±

4.31c

98.64±

0.12 d

88.35±

2.02 d

80.81±

0.78 c

79.00±

1.44 c

21.60±

4.38b

99.72±

0.25d

94.08±

2.90f

89.52±

1.18c

83.81±

1.28c

83.52

L.S.D0.95 2.9791 6.9930 9.3038 28.2074 10.9737 8.1949 3.9962 8.8272 8.10106 12.9775 10.4526 5.9390 9.6575 9.8823

* the second spray. ** the third spray. S.D: Standard deviation. Means followed by the same letter (s) are not significantly difference (P= 0.95 level)

141


0 1 3 5 7 9 10 12 14 16 18 19 21 23 25

Time (days).

0

5

10

15

20

Av e

rage

num

ber

of i n

sect

s/3

lea v

es

pirimiphos-methyl pirimicarb propoxur dimethoate chlorpyrifos-methyl

phenthoate profenofos carbosulfan Control.

Fig. (10) :The average number of larvae of the cotton leafworm Spodoptera littoralis / 3 leaves infesting okra plants.

142


respectively). Although pirimiphos-methyl, carbosulfan and phenthoate

exhibited a reasonable initial efficiency (% reduction after 24 hours were

80.11, 78.34 and 75.29, respectively) but their effects were greatly

decreased after 3 days of spraying (% reduction: 47.15, 5.13 and 59.25,

respectively). In general, the vegetable crops are commonly infested and

attacked by several insect species particularly those belonging to order

Lepidoptera (e.g. S. littoralis). El- Dahan (1991) studied the survey of

insecticide resistance in the cotton leafworm, S. littoralis which was

collected from Kafr El-sheikh Governorate during the years 1984-1988.

The results indicated that, field strains developed high resistance level to

pyrethroids while the organophosphates, chloropyrifos and profenofos

were more effective. It is worthwhile to mention that, chloropyrifos-

methyl and profenofos are still officially recommended to control S.

littoralis on vegetable plants (National program of Agricultural pests

control, 1997, Ministry of Agriculture and Land Reclamation, Egypt).

1.2.2 The whitefly, Bemisia tabaci :

Okra and squash plants were heavily infested with the whitefly, B.

tabaci. The insect was abundantly existed on plants throughout the

examination period which lasted more than three weeks. For okra plants

results were recorded in Table (11-18) and illustrated in Fig 11-14.

From the obtained results, it is clear that, apart from pirimiphos-

methyl, profenofos and carbosulfan whose efficiency were relatively

moderate (Average % reduction ranged 64-78), the tested insecticides

were of poor efficacy. The insecticides retained their moderate toxicities

up to 9 days after spraying. On squash plants, pirimiphos-methyl,

profenofos and phenthoate were moderately effective. Although

chloropyrifos-methyl was poorly effective against whitefly on okra plants

(average of % reduction: 31.49 and 35.56 for adult and immature stage,

143


respectively), it was of relatively high effect on immature stages of

whitefly infesting squash plants(average % reduction: 83.13). Results

showed also that, the tested insecticides varied in their efficiency

depending on the tested stage ( nymph or adult) and the type of plants

carrying the insect (i.e. okra or squash). The obtained results agreed with

those of many authors. Zeitoun et al (1990) reported that profenofos at

rate of 750 ml/F. was potent against B. tabaci on potatoes with an average

reduction of 85% . Chloropyrifos-methyl gave acceptable control to

whitefly on cucumber plants (El- Sayed and El-Ghar, 1992; Abbassy et

al, 1993). Pirimiphos-methyl achieved a good control against white flies

infesting cucumber plants in plastic houses (Abbassy et al. 1993; Zidan,

et al. 1994). On cotton plants, carbosulfan gave acceptable control to B.

tabaci (Rizk and Kamel, 1991). However, carbosulfan and pirimiphos-

methyl failed to give effective control against the whitefly on cabbage

plants (Percentages of reduction were 41 and 42 for carbosulfan and

pirimiphos-methyl, respectively) (Farrag et al. 1994). El- Maghraby, et

al. (1997) found that profenofos, pirimiphos-methyl and chloropyrifos-

methyl successfully reduced the population of various stages of the

whitefly, B. tabaci on tomato plants.

Generally, the cotton whitefly, B. tabaci is considered one of the most

serious pests attacking plants. In Egypt, it is well known as a key pest on

vegetable crops especially tomatoes and squash in traditional or protected

plantations. Some investigators attributed the increase of whitefly in the

last few years to changes in ecosystem such as intensification and

diversitification of the cropping system which has provided more hosts

and the use of broad spectrum and persistent chemicals which suppressed

the natural enemies (Darwish and Farghal 1990). On the other hand,

144

RESULTS AND DISCUSSIONTable (11): Efficiency of certain pesticides against the population density of nymphs of cotton whitefly, Bemisia tabaci in okra plants .

Chemical Average No. of nymphs/3 leaves ± S.D. at days:-used Before

1 st

spray.

1 3 5 7 9* 10 12 14 16 18** 19 21 23 25

Carbosulfan 18.94±

0.48

2.77±

0.17

4.73±

0.17

5.71±

0.20

9.88±

0.29

11.30±

0.05

1.21±

0.14

2.37±

0.09

3.01±

0.15

4.35±

0.21

5.51±

0.34

0.54±

0.10

1.08±

0.09

1.00±

0.03

1.42±

0.03

Pirimicarb 19.40±

1.13

10.38±

0.19

13.27±

0.72

16.72±

0.38

20.56±

0.44

23.89±

0.75

13.74±

0.65

13.76±

0.32

15.96±

0.52

20.60±

0.47

25.94±

0.50

14.45±

0.42

14.46±

0.92

12.81±

0.33

15.95±

0.46

Propoxur 12.69±

0.85

5.21±

0.20

7.44±

0.06

8.55±

0.27

16.19±

0.64

19.55±

0.79

7.77±

0.67

9.76±

0.54

12.07±

0.68

19.68±

1.05

22.20±

1.41

8.69±

0.31

8.75±

0.41

7.62±

0.22

14.77±

0.19

Chlorpyrifos-methyl

15.40±

0.73

7.73±

0.24

10.58±

0.36

13.34±

0.53

16.88±

0.29

20.61±

0.75

9.25±

0.10

11.68±

0.77

14.51±

0.40

18.55±

0.82

22.15±

0.04

9.06±

0.50

9.66±

0.17

8.52±

0.23

10.38±

0.13

Dimethoate 10.69±

0.58

2.71±

0.20

5.52±

0.32

6.20±

0.23

13.96±

0.32

16.65±

0.86

3.44±

0.25

6.65±

0.99

10.41±

2.44

16.40±

0.57

18.00±

1.79

4.27±

0.32

6.34±

0.27

4.42±

0.26

10.63±

0.21

Phenthoate 11.01±

0.53

6.45±

0.44

8.70±

0.21

12.21±

0.32

15.46±

0.16

17.90±

0.58

9.18±

0.19

11.77±

0.46

14.67±

0.74

18.80±

0.16

21.71±

0.66

12.54±

0.21

12.39±

0.23

11.43±

0.16

14.44±

0.23

Pirimiphos-methyl

26.31±

1.59

0.96 ±

0.09

3.88 ±

0.29

9.17 ±

0.51

12.43 ±

0.07

14.09 ±

0.59

0.16 ±

0.01

2.21 ±

0.22

4.26 ±

0.21

5.01 ±

0.15

6.60 ±

0.28

0.03 ±

0.001

0.94 ±

0.04

1.20 ±

0.04

1.47 ±

0.21

Profenofos 10.78±

0.30

1.36±

0.29

1.69±

0.18

2.56±

0.08

3.51±

0.25

4.30±

0.39

0.50±

0.12

0.74±

0.03

1.05±

0.20

1.56±

0.07

2.56±

0.29

0.23±

0.01

0.38±

0.04

0.35±

0.04

0.47±

0.02

Control 14.68±

0.18

12.56±

0.29

15.91±

0.32

18.76±

0.27

23.43±

0.78

25.82±

0.66

20.56±

0.78

22.87±

0.29

25.83±

0.40

31.10±

0.79

33.65±

0.75

29.29±

0.28

27.73±

2.10

22.25±

0.72

25.88±

0.53

the second spray (population density just before sprying) ,** the third spray (population density just before sprying) . S.D: Standard deviation.

145


Table ( 12 ): Efficiency of certain pesticides against nymphs of cotton whitefly, Bemisia tabaci i n okra plants



1.49 e

76.93±

1.17 e

76.39±

0.33ef

67.3±

1.60 c

66.06±

0.91d

86.59±

1.49 d

76.32±

1.01 e

73.38±

1.17 e

68.07±

1.40 d

62.57±

2.47d

88.73±

2.51 f

76.09±

3.27 d

72.36±

2.56 f

66.31±

2.42 e

74.28

Pirimicarb 500 gm. 37.30±

3.20 b

36.61±

6.52 b

32.31±

5.55 b

33.39±

5.35 b

29.84±

4.40c

27.73±

2.58 a

34.90±

2.97 b

33.16±

3.14 b

28.35±

2.73 c

16.65±

1.19b

36.00±

0.62 b

32.29±

5.57 b

25.3±

3.11 b

20.03±

1.54 c

30.27

Propoxur 1 L. 51.75±

4.92 c

45.75±

3.20 c

47.03±

4.89 c

19.71±

8.33 a

12.33±

2.68a

50.13±

2.84d

55.04±

4.24ef

38.28±

0.97b

16.43±

1.75b

12.89±

2.05ab

54.88±

3.94b

51.98±

4.69b

47.93±

4.31d

13.27±

4.62a

36.96

Chlorpyrifos-methyl

1 L. 41.2±

3.76 b

36.54±

2.73 b

32.09±

4.31 b

31.21±

3.73 b

23.83±

3.03b

43.60±

1.88c

35.99±

4.85d

29.60±

2.20b

25.29±

1.80c

17.46±

2.83b

53.02±

2.53b

47.10±

0.86b

41.85±

1.58c

39.05±

0.87c

35.56

Dimethoate 1L. 70.23±

3.00 d

52.28±

1.58 d

54.5±

2.80 d

17.97±

5.71 a

11.32±

5.06a

74.02±

1.03e

43.64±

4.25c

37.22±

16.25b

18.12±

2.49b

17.17±

3.84b

72.45±

4.29c

56.97±

4.34c

62.67±

3.00e

22.66±

7.64b

43.66


3.53 a

27.01±

2.76 a

12.96±

6.15 a

11.85±

5.07 a

7.51±

1.5b

35.55±

1.34b

25.79±

0.86b

18.10±

1.54a

12.73±

2.57a

6.94±

0.28a

33.57±

2.86a

30.69±

3.34a

20.31±

3.40a

13.48±

1.42a

20.57

Pirimiphos-methyl

1.5 L. 95.70±

0.63 f

86.35±

1.51 f

72.62±

2.74 e

70.32±

1.69 c

69.51 ±

0.887d

98.56±

0.05 e

82.3±

1.50 f

69.69±

2.68 e

70.41±

1.99 d

64.03 ±

1.22d

99.41±

0.047g

82.67±

0.13 e

72.46±

2.04 f

70.89±

4.93 f

78.92


3.56 e

85.55±

1.20 f

81.37±

1.11 f

79.61±

1.55 d

77.36±

1.60e

85.33±

3.98f

80.40±

1.11a

75.72±

2.52c

69.71±

1.48d

53.77±

9.65c

89.47±

1.04 f

81.91±

1.53 e

79.49±

0.44 g

75.76±

2.39 g

78.62

L.S.D 0.95 5.6527 5.3161 6.9177 8.1661 5.0488 3.8346 4.7029 10.5491 3.6093 6.927 4.5931 6.0202 4.8304 6.7199

* the second spray . ** the third spray . S.D: Standard deviation. Means followed by the same letter (s) are not significantly difference (P= 0.95 level).

146


0 1 3 5 7 9 01 21 41 61 81 91 12 32 52

.(syad) emiT

0

5

01

51

02

52

03

53

s eva

el 3

/stc

e sni

fo

r ebm

un e

gare

v A

lyhtem-sohpimirip bracimirip ruxoporp etaohtemidlyhtem-sofiryprolhc

etaohtnehp sofoneforp naflusobrac .lortnoC

Fig.(11): The average number of immature stage of whitefly Bemisia tabaci/ 3 leaves infesting okra plants.

147


Table (13): Efficiency of certain pesticides against the population density of the adults of cotton whitefly, Bemisia tabaci in okra plants

.

Chemical No of adults whitefly/3 leaves ± S.D. at days:-used Before

1st

spray.

1 3 5 7 9* 10 12 14 16 18** 19 21 23 25

Carbosulfan 26.45±

2.53

3.57±

0.35

3.44±

0.15

5.67±

0.21

7.39±

0.44

8.51±

0.34

0.72±

0.11

0.84±

0.11

0.95±

0.09

1.73±

0.18

2.58±

0.12

0.27±

0.03

0.44±

0.05

0.97±

0.06

1.66±

0.08

Pirimicarb 13.90±

0.88

3.45±

0.15

5.34±

0.29

6.45±

0.19

4.39±

0.34

5.61±

0.28

1.09±

0.04

1.67±

0.27

1.25±

0.14

1.69±

0.21

2.59±

0.20

0.62±

0.03

1.35±

0.05

2.43±

0.18

3.32±

0.2 b

Propoxur 8.61±

0.15

4.53±

0.18

5.57±

0.21

8.10±

0.09

5.33±

0.29

6.74±

0.14

2.03±

0.05

3.54±

0.18

3.67±

0.27

4.11±

0.12

5.150±

0.13

2.30±

0.16

4.59±

0.20

8.39±

0.12

9.13±

0.39

Chlorpyrifos-methyl

11.25±

0.29

4.62±

0.26

5.43±

0.09

11.27±

0.26

7.07±

0.08

8.53±

0.34

1.92±

0.07

3.06±

0.06

4.48±

0.23

5.30±

0.14

6.69±

0.26

2.31±

0.17

4.56±

0.19

11.28±

0.22

11.55±

0.39

Dimethoate 7.62±

0.36

0.74±

0.05

0.67±

0.04

3.71±

0.20

3.51±

0.24

3.74±

0.1 d

0.40±

0.23

0.32±

0.09

1.16±

0.05

1.95±

0.07

2.54±

0.12

0.15±

0.02

0.25±

0.01

2.07±

0.12

3.54±

0.32

Phenthoate 17.43±

0.45

2.65±

0.18

3.60±

0.36

5.23±

0.22

9.70±

0.17

10.22±

0.26

1.14±

0.05

1.61±

0.14

1.71±

0.18

5.57±

0.10

6.80±

0.32

0.87±

0.06

1.62±

0.18

3.22±

0.19

10.74±

0.34

Pirimiphos-methyl

9.72 ±

0.46

2.24±

0.15

1.83 ±

0.06

3.02 ±

0.08

3.49 ±

0.43

4.57 ±

0.32

0.54 ±

0.05

0.84 ±

0.02

0.93 ±

0.09

1.60 ±

0.18

2.47 ±

0.42

0.44 ±

0.12

0.64 ±

0.19

1.41 ±

0.25

2.81 ±

0.58

Profenofos 16.56±

0.39

2.27±

0.26

2.28±

0.02

4.57±

0.23

4.52±

0.24

4.67±

0.19

0.34±

0.10

0.57±

0.10

0.80±

0.11

1.49±

0.12

2.05±

0.10

0.15±

0.03

0.45±

0.19

1.09±

0.12

1.75±

0.28

Control 36.68±

0.82

45.37±

0.28

35.78±

0.39

39.27±

0.73

24.52±

0.40

24.46±

0.15

28.85±

0.65

19.97±

0.52

16.83±

1.43

18.25±

0.64

18.82±

0.11

20.56±

0.68

24.56±

0.95

34.67±

0.97

36.18±

2.34


148


Table ( 14 ): Efficiency of certain pesticides against adults of cotton whitefly, Bemisia tabaci in okra plants.

Chemical Rate/ Average % reduction ± S.D. at days:- Averageused fed 1 3 5 7 9* 10 12 14 16 18** 19 21 23 25 reduction


1.72 d

86.57±

1.77 e

79.80±

2.57 e

57.95±

5.03 e

51.48±

4.82f

92.85±

0.80 e

87.82±

1.93 f

83.66±

2.08 d

72.74±

1.93 f

60.52±

3.19e

90.41±

0.99 d

86.78±

1.95 fig

79.66±

0.40 d

66.45±

2.81 d

77.54


0.61 c

60.53±

3.40 c

56.6±

2.4 c

52.75±

1.27 de

39.17±

6.57c

83.57±

0.58 c

63.68±

4.38 c

67.76±

2.42 c

59.74±

3.39 e

40.07±

4.59d

78.1±

1.73c

59.78±

2.58c

49.05±

0.23b

33.29±

0.79d

58.82


2.30 a

33.66±

2.46 a

12.13±

2.54 b

7.38±

3.54 a0.00 a

74.42±

0.74 a

35.79±

2.10 a

20.96±

4.72 a

18.29±

4.00 a0.00 a

59.05±

1.87 a

31.64±

4.23 a

11.43±

1.00 a

7.62±

5.64 a

26.42

Chlorpyrifos-methyl

1 L. 66.82±

1.86 b

50.48±

0.89 b

6.4±

3.85 a

6.01±

3.31 a0.00 a

80.88±

1.33 b

56.04±

2.13 b

23.38±

6.85 a

16.61±

1.34 a0.00 a

68.39±

1.37 b

47.62±

3.58 b

8.31±

4.88 a

9.95±

6.57 a

31.49


0.42 e

90.94±

0.51 f

54.46±

3.12 c

30.85±

6.82 c

26.17±

5.58c

90.97±

5.35c

89.47±

2.71f

54.93±

1.52b

29.91±

5.40b

11.82±

2.36b

94.47±

0.72 d

92.33±

0.70 g

55.73±

3.74 bc

27.36±

7.65 ab

60.11


0.80 d

78.83±

2.37 d

71.95±

1.41 d

16.72±

1.73 b

11.98±

4.29b

90.54±

0.60 d

80.76±

1.38 de

75.67±

1.98 c

26.93±

0.51 b

13.5±

6.05b

88.26±

1.032e

81.74±

2.24d

74.30±

1.13d

17.71±

2.987ab

58.32

Pirimiphos-methyl

1.5 L. 81.39±

0.80 c

80.66±

1.24 d

70.95±

0.72 d

45.95±

9.04 d

29.16±

8.17c

89.93±

0.14 d

77.32±

2.12 d

70.21±

4.97 c

53.08±

4.54 d

30.05 ±

7.03c

84.01±

1.54d

80.46±

2.62d

68.95±

4.07ec

40.71±

7.47d

64.48


1.51 d

85.90±

0.42 e

74.18±

1.88 d

59.13±

2.43 e

57.69±

2.08f

93.93±

1.50c

80.00±

2.04ff

75.17±

3.58c

26.93±

0.51c

42.90±

3.45d

93.32±

0.99 d

83.21±

2.70 ef

71.19±

1.95 d

55.37±

9.28 cd

70.56

L.S.D 0.95 2.4354 3.3027 4.3055 8.37924 9.9334 3.5984 4.3191 6.7884 5.9851 8.4309 2.3118 4.776 4.766 10.4726

* the second spray . ** the third spray. S.D: Standard deviation. Means followed by the same letter (s) are not significantly difference (P= 0.95 level) .

149


0 1 3 5 7 9 10 12 14 16 18 19 21 23 25

Time (days).

0

10

20

30

40

50

Av e

rage

num

ber

of i n

sect

s/3

lea v

es



Fig. (12):The average number of adult of whitefly Bemisia tabaci / 3 leaves infesting okra plants.

150

RESULTS AND DISCUSSIONTable (15 ): Efficiency of certain pesticides against the population density of nymphs of cotton whitefly, Bemisia tabaci in squash plants .

Chemical No of nymphs /leaf ± S.D. at days:-used Before

1 st

spray.

1 3 5 7 9* 10 12 14 16 18** 19 21 23 25

Carbosulfan 18.88±

0.72

0.01±

0.01

2.46±

0.32

17.35±

0.34

13.93±

0.50

14.71±

0.63

14.71±

0.63

2.66±

0.23

8.21±

0.21

11.45±

0.20

12.39±

0.53

0.16±

0.02

1.61±

0.25

7.21±

0.05

8.46±

0.32

Pirimicarb 23.78±

0.71

8.55±

0.37

10.95±

0.44

25.31±

1.13

17.32±

0.85

18.80±

0.71

6.60±

0.24

9.69±

0.60

12.35±

0.20

16.34±

0.20

17.15±

1.19

4.00±

0.70

6.28±

0.20

10.47±

0.24

11.53±

0.57

Propoxur 37.34±

1.03

17.39±

2.13

29.67±

0.32

53.14±

0.93

37.23±

1.14

38.89±

0.57

19.44±

0.44

33.58±

0.40

33.71±

0.51

43.95±

0.62

44.79±

0.87

17.80±

0.31

27.05±

0.55

37.57±

1.37

42.25±

0.30

Chlorpyrifos-methyl

44.27±

2.23

3.12±

0.14

7.26±

0.27

14.78±

0.48

11.91±

0.85

13.41±

0.50

0.64±

0.02

1.82±

0.25

2.74±

0.26

4.51±

0.24

4.89±

0.24

0.22±

0.09

0.67±

0.14

1.04±

0.02

1.29±

0.11

Dimethoate 34.65±

1.15

5.55±

0.37

15.03±

0.95

29.59±

0.57

25.61±

0.90

26.94±

0.33

4.10±

0.95

11.63±

0.74

12.79±

0.71

20.70±

0.25

22.64±

0.52

3.61±

0.29

8.06±

0.29

11.51±

0.16

15.48±

0.45

Phenthoate 48.70±

0.52

0.52±

0.04

8.42±

0.04

25.34±

1.17

23.84±

0.37

25.08±

0.31

25.08±

0.31

5.10±

0.08

9.41±

0.15

14.11±

0.61

15.75±

0.70

0.16±

0.09

1.86±

0.08

4.83±

0.11

7.26±

0.75

Pirimiphos-methyl

36.44±

0.84

7.09±

0.12

9.51 ±

0.08

38.69 ±

0.89

26.65 ±

0.70

27.90 ±

0.51

5.24 ±

0.20

8.63±

0.28

18.80±

0.63

23.86±

0.62

25.50 ±

1.01

3.03 ±

0.12

5.23 ±

0.23

17.17 ±

0.63

17.51 ±

0.33

Profenofos 22.51±

1.09

0.01±

0.01

4.60±

0.35

8.61±

0.29

10.67±

0.30

11.68±

0.72

11.68±

0.72

2.32±

0.11

2.72±

0.29

5.52±

0.36

6.06±

0.44

0.05±

0.03

1.00±

0.14

1.59±

0.16

2.71±

0.14

Control 33.05±

0.50

36.83±

1.17

39.45±

0.67

59.75±

0.35

41.01±

0.73

40.67±

0.75

40.67±

0.75

49.38±

1.07

43.44±

0.95

55.01±

1.98

54.61±

1.44

49.21±

0.52

48.77±

0.34

58.05±

0.49

60.28±

0.90

the second spray (population density just before sprying) ,** the third spray (population density just before sprying). S.D: Standard deviation.

151


Table ( 16): Efficiency of certain pesticides against nymphs of cotton whitefly, Bemisia tabaci in squash plants.



0.05 e

89.05±

1.73 e

49.10±

2.61 c

40.43±

4.11 b

36.67±

2.08 b

99.40±

0.08 f

85.05±

1.87 e

47.69±

2.20 c

42.38±

2.31 c

37.19±

3.96 b

98.56±

0.23 e

85.37±

2.90 d

45.20±

2.62 c

38.00±

5.06 b

63.26


2.18 b

61.39±

1.46 b

41.03±

4.28 b

41.27±

3.41 b

35.72±

3.01 b

68.80±

0.23 b

57.46±

4.01 b

38.43±

2.96 b

35.70±

1.90 b

32.11±

2.17 b

73.99±

5.15 b

58.86±

3.74 b

42.30±

5.19 c

38.83±

6.38 b

49.54


6.28 a

33.37±

2.55 a

21.2±

3.54 a

19.54±

4.70 a

15.32±

1.29 a

55.55±

1.63 a

28.87±

1.82 a

18.83±

2.02 a

16.42±

2.38 a

14.23±

0.74 a

55.88±

0.83 a

32.37±

1.56 a

21.04±

4.14 a

14.53±

1.04 a

28.95

Chlorpyrifos-methyl

1 L. 93.67±

0.06 d

86.26±

0.37 de

81.51±

0.76 e

78.28±

1.91 d

75.33±

1.83 b

95.77±

0.11 e

88.76±

1.86 f

80.79±

2.45 e

75.14±

1.39 f

72.80±

2.21 e

94.97±

1.89 e

84.00±

3.98 d

79.90±

0.63 f

76.05±

2.27 d

83.13

Dimethoate 1 L. 85.63±

0.50 c

63.6±

3.23 b

52.73±

1.99 c

40.42±

0.97 b

36.75±

2.01 b

86.51±

2.99 c

64.48±

1.86 c

55.56±

2.11 d

43.19±

1.02 c

37.43±

1.51

82.27±

1.85 c

60.16±

0.75 b

52.13±

1.26 d

38.04±

1.69 b

57.06


0.08 e

85.52±

0.10 de

71.21±

1.45 d

60.54±

0.82 c

58.15±

0.75 c

99.97±

0.023 f

83.26±

0.45 e

64.85±

0.97 d

58.39±

2.18 d

53.21±

2.62 c

98.1±

0.62 e

86.74±

0.43 d

71.10±

1.53 e

58.25±

3.99 c

74.88

Pirimiphos-methyl

1.5 L. 82.53±

0.70 c

78.14±

0.33 c

41.22±

2.67 b

41.00±

2.88 b

37.77±

1.18 b

83.32±

0.52 c

74.53±

1.06 d

36.89±

2.55 b

36.75±

2.07 b

31.94 ±

1.96 b

86.79±

1.00 d

76.97±

1.89 c

36.54±

4.75 b

37.72±

3.32 b

55.865


0.02 e

82.84±

1.92 d

78.79±

1.72 e

61.69±

2.94 c

57.84±

0.64 c

98.55±

0.16 f

83.61±

0.52 e

78.08±.

3.66 e

64.87±

4.37 e

61.19±

5.18 d

98.99±

0.67 e

81.51±

1.167 d

75.3±

1.85 ef

59.47±

0.89 c

77.34

L.S.D 0.95 4.1002 3.122 4.5110 5.2239 3.0477 2.1212 3.4294 4.2850 4.1378 4.9611 3.6778 4.1636 5.5156 6.2153

* the second spray . ** the third spray . S.D: Standard deviation. Means followed by the same letter (s) are not significantly difference (P= 0.95 level).

152


0 1 3 5 7 9 01 21 41 61 81 91 12 32 52

.(syad) emiT

0

01

02

03

04

05

06

07

fael

/stc

e sni

fo

r ebm

un e

gare

v A



Fig. (13):The average number of immature stage of Bemisia tabaci/ leaf infesting squash plants.

153

RESULTS AND DISCUSSIONTable (17): Efficiency of certain pesticides against the population density of adults of cotton whitefly, Bemisia tabaci in squash plants

Chemical Average no.of adults whitefly/leaf ± S.D. at days:-used Before

spray.1 3 5 7 9* 10 12 14 16 18** 19 21 23 25

Carbosulfan 6.63±

0.30

5.55±

0.31

6.66±

0.10

9.24±

0.18

8.25±

0.24

10.46±

0.36

5.63±

0.18

5.73±

0.28

6.58±

0.13

7.52±

0.42

8.49±

0.28

5.67±

0.25

4.76±

0.16

5.70±

0.19

4.88±

0.12

Pirimicarb 5.49±

0.32

3.53±

0.11

6.38±

0.46

9.63±

0.21

9.08±

0.09

10.65±

0.34

4.60±

0.37

7.22±

0.19

8.59±

0.35

10.26±

0.53

12.00±

0.17

5.94±

0.49

8.87±

0.28

10.42±

0.15

9.73±

0.16

Propoxur 4.57±

0.33

2.56±

0.21

4.28±

0.36

5.66±

0.16

5.50±

0.19

7.32±

0.44

2.64±

0.29

3.78±

0.05

4.45±

0.15

5.43±

0.43

6.62±

0.24

2.79±

0.28

3.55±

0.04

4.05±

0.18

4.09±

0.40

Chlorpyrifos-methyl

4.57±

0.33

1.87±

0.08

5.43±

0.09

8.46±

0.13

7.69±

0.20

8.86±

0.42

2.11±

0.14

5.71±

0.27

7.50±

0.45

9.07±

0.45

9.97±

0.29

2.64±

0.12

6.58±

0.14

8.91±

0.18

8.44±

0.31

Dimethoate 3.68±

0.27

1.71±

0.14

4.53±

0.32

6.64±

0.15

6.25±

0.20

6.99±

0.44

2.26±

0.11

4.69±

0.27

5.70±

0.23

7.25±

0.51

8.48±

0.08

3.48±

0.17

6.32±

0.24

7.33±

0.19

7.23±

0.22

Phenthoate 7.43±

0.18

1.05±

0.04

3.49±

0.24

7.12±

0.12

7.13±

0.13

10.40±

0.54

0.96±

0.30

2.47±

0.11

4.40±

0.09

5.62±

0.15

8.02±

0.42

0.42±

0.13

1.79±

0.06

3.47±

0.33

3.40±

0.24

Pirimiphos-methyl

4.32 ±

0.12

0.46±

0.06

0.97±

0.08

1.57 ±

0.28

1.78±

0.26

2.28±

0.22

0.16 ±

0.01

0.32 ±

0.03

0.61 ±

0.02

0.62 ±

0.07

0.85 ±

0.05

0.05 ±

0.01

0.15 ±

0.01

0.24 ±

0.04

0.23±

0.02

Profenofos 3.97±

0.13

2.44±

0.16

3.35±

0.16

5.06±

0.05

4.49±

0.22

6.56±

0.13

2.51±

0.07

2.89±

0.18

3.80±

0.19

4.62±

0.20

6.53±

0.05

2.73±

0.24

3.33±

0.10

3.26±

0.12

3.65±

0.25

Control 5.00±

0.22

6.51±

0.07

8.22±

0.22

10.44±

0.07

9.31±

0.19

10.45±

0.71

9.46±

0.11

9.89±

0.23

10.29±

0.14

11.54±

0.37

12.43±

0.38

13.68±

0.25

12.59±

0.26

13.04±

0.36

11.47±

0.37


154


Table ( 18 ): Efficiency of certain pesticides against adults of cotton whitefly, Bemisia tabaci in squash plants .



2.53 a

38.82±

1.89 b

33.16±

2.02 b

33.13±

2.48 b

24.42±

2.76b

40.47±

3.87a

42.02±

4.78b

36.08±

3.41b

34.81±

5.33c

31.76±

3.67e

39.14±

4.56 a

44.62±

1.99 c

35.95±

4.02 b

37.64±

3.27 b

36.28


3.73 b

28.96±

8.68 a

15.73±

6.35 a

10.97±

4.58 a

7.04±

3.50a

52.36±

2.32b

28.38±

1.11a

18.153±

1.51a

12.83±

1.75b

5.196±

3.89b

55.16±

4.19b

27.06±

3.06a

17.26±

1.64a

12.09±

1.611a

24.39


5.83 b

42.97±

3.75 b

40.46±

5.35 b

35.33±

2.49 b

23.32±

2.48b

60.18±

2.12c

45.38±

2.56b

38.26±

2.02bc

32.87±

1.67c

23.92±

1.76d

61.80±

2.57c

47.06±

2.29cd

41.65±

0.86c

33.12±

4.08b

41.65

Chlorpyrifos-methyl

1 L. 68.35±

1.84 c

26.99±

5.45 a

10.41±

7.23 a

8.75±

8.01 a

6.50±

3.91a

73.68±

1.37e

31.93±

1.56a

14.13±

1.39a

7.3±

0.96a

5.33±

2.61b

75.95±

0.43d

34.85±

2.06b

14.80±

2.07a

7.64±

3.03a

27.62


0.59 c

24.68±

10.19 a

13.153±

7.77 a

8.49±

5.11 a

9.05±

2.18a

64.29±

1.4d

29.05±

0.49a

17.14±

1.92a

6.01±

1.25a0.00 a

62.71±

2.08 c

26.50±

3.02 a

17.62±

1.97 a

7.64±

3.04 a

25.05


0.49 d

71.47±

2.21 b

54.12±

1.77 c

48.51±

0.81 c

33.00±

5.14c

89.92±

2.64f

74.90±

1.63d

56.97±

1.30d

51.02±

1.22d

35.16±

0.98e

95.16±

1.67e

77.97±

1.58e

58.7±

4.67e

8.27±

0.99c

60.31

Pirimiphos-methyl

1.5 L. 91.88±

0.98 d

86.28±

1.45 d

82.52±

3.42 d

77.82±

3.80 d

74.75 ±

1.73d

92.19±

1.16 f

85.21±

2.34 e

72.91±

1.92 e

75.65±

4.67 e

68.66 ±

3.39f

94.28±

0.43 e

82.52±

0.26 f

72.89±

5.39 f

70.38±

4.39 d

80.57


3.71 b

48.70±

3.33 c

38.92±

2.60 b

39.34±

2.40 b

20.68±

2.59b

57.79±

1.47c

53.61±

3.49c

41.33±

2.46c

36.49±

1.56c

16.57±

1.72c

62.01±

3.19 c

49.62±

1.22 d

52.37±

1.72 d

39.46±

4.19 b

43.54

L.S.D 0.95 5.2280 9.5721 8.8129 7.3516 3.8298 3.8298 4.4842 3.6268 4.8393 4.8586 4.8423 3.6823 5.5301 5.5289

* the second spray . ** the third spray . S.D: Standard deviation. Means followed by the same letter (s) are not significantly difference (P= 0.95 level)

155


0 1 3 5 7 9 01 21 41 61 81 91 12 32 52

.(syad) emiT

0

2

4

6

8

01

21

41

61

fael

/stc

e sni

fo

r ebm

un e

gare

v A



Fig .(14):The average number of adult of whitefly Bemisia tabaci / leaf infesting squash plants.

156


due to the extensive applications of pesticides against whitefly, it has

become resistant to many group of insecticides (Prodbhaker, et al.

1985). Therefore, chemical control of whitefly have been tried in many

parts of the world with variable degrees of success.

1.2.3. Aphids, A. gossypii :

Aphid, A. gossypii was observed only on squash plants and was not

observed on okra plants through the experimental period. Results

pertaining the efficiency of the tested insecticides against the aphids were

recorded in Table 19, 20 and illustrated in Fig.15. From the results

obtained it is evident that, the population of aphids started with low

numbers then increased rapidly to reach a maximum of 62 aphids/leaf of

squash plant throughout the experimental period. Carbosulfan was the

most effective insecticide followed by chloropyrifos-methyl and

pirimicarb (average percents of reduction through test period: 87.69, 84.7

and 82.86). The least effective compounds were pirimiphos-methyl and

propoxur (% reduction: 60.29 and 56.4, respectively). However, all tested

insecticides resulted in a considerable reduction after 24 hours of

spraying. The effectiveness of the insecticides, carbosulfan,

chloropyrifos-methyl and pirimicarb lasted up to 9 days after spraying

whereas that of the rest of

insecticides were reduced after 3 days. The results of the present study

are in agreement with those of many authors. Shalaby et al. (1991) found

that Marshal(carbosulfan) 25% at rate of 300 gm/fed gave exellent control

for aphids. Against the cabbage aphid Brevicoryne brassicae,

chloropyrifos-methyl was the most toxic insecticide among other

pesticides (El-Sayed et al. 1991). Also Rizk and Kamel (1991) found

that, chloropyrifos methyl bioencapsulated at a rate of 400 gm/F.

produced an immediate effect on aphids after 3 days of application in

157


comparison with chlorpyrifos-methyl (EC). The specificaphicidal action

of pirimicarb against different genera and species of aphids were

currently reported(Zidan et al. 1988a and 1989b ; El- Defrawi and Abd

El-Azim, 1992; Abo El- Ghar et al. 1993).

Generally, the aphid control in vegetable and field crops has

become more difficult in recent years. Massive application of insecticides

have led to resistance phenomenon and depleted the aphid parasitoids and

predators. Table (21) shows that various insect pests can coexist on the

same crop (i.e aphids and whitefly on squash; the cotton leafworm and

whitefly on okra plants). This coincidence of insect species may be of

great importance from the chemical control point of view. No doubt that,

controlling more than one type of pests simultaneously might reduce

numbers of sprays and thus reducing hazards of pesticides used as

possible.

Based on the obtained results of laboratory and field studies, it is

evident that pirimicarb and chloropyrifos-methyl have many advantages

enough to make them the most promising among the tested insecticides

for application on vegetable plants for two reasons:

1-Chloropyrifos-methyl had the highest toxicity against aphids

taking in consideration the three tested species of aphids( as shown in the

laboratory tests). Meanwhile, chlorpyrifos-methyl was the most toxic

compound to the cotton leafworm S. littoralis. Pirimicarb was among the

insecticides that exhibited relatively reasonable toxicity to aphids. In field

studies, chloropyrifos-methyl and pirimicarb still having a high efficiency

against aphid, A. gossypii on squash plants. On squash plant also

chlorpyrifos-methyl was of relatively high effect on immature stages of

the whitefly, B. tabaci and the compound lasted effective up to 9 days

after spraying. On okra plants, chloropyrifos-methyl exhibited a

158

RESULTS AND DISCUSSIONTable (19): Efficiency of certain pesticides against the cotton aphid, Aphis gossypii in squash plants.

Chemical Average no.of aphids/leaf ± S.D. at days:-used Before

1 st

spray.

1 3 5 7 9* 10 12 14 16 18** 19 21 23 25

Carbosulfan 3.25±

0.29

0.01±

0.01

0.02±

0.01

0.02±

0.01

0.23±

0.05

1.45±

0.13

0.01±

0.01

0.02±

0.01

0.06±

0.02

0.13±

0.64

0.32±

0.04

0.04±

0.01

0.15±

0.03

0.44±

0.07

0.68±

0.18

Pirimicarb 3.45±

0.30

0.12 ±

0.02

0.50±

0.09

1.14±

0.25

1.35±

0.10

2.51±

0.36

0.05±

0.01

0.41±

0.06

0.51±

0.08

0.94±

0.07

1.74±

0.21

0.21±

0.25

0.53±

0.11

0.68±

0.06

0.96±

0.11

Propoxur 2.59±

0.38

0.66±

0.07

0.91±

0.07

1.48±

0.26

2.46±

0.38

4.35±

0.48

1.37±

0.38

1.41±

0.37

3.42±

0.18

4.03±

0.52

5.99±

0.21

3.21±

0.28

3.00±

0.19

9.32±

1.12

13.52±

1.11

Chlorpyrifos-methyl.

2.48±

0.41

0.01±

0.01

0.06±

0.01

0.37±

0.05

0.48±

0.07

1.46±

0.38

0.01±

0.01

0.01±

0.01

0.24±

0.05

0.33±

0.08

0.51±

0.09

0.00±

0.00

0.01±

0.01

0.16±

0.08

0.18±

0.14

Dimethoate 2.86±

0.36

0.16±

0.02

0.46±

0.04

1.43±

0.42

1.78±

0.21

3.94±

0.15

0.05±

0.01

0.48±

0.05

1.81±

0.19

2.63±

0.35

3.36±

0.21

0.12±

0.07

0.73±

0.3.9

3.86±

00.6

4.00±

0.47

Phenthoate 2.67±

0.71

0.13±

0.02

0.74±

0.03

0.77±

0.05

1.35±

0.07

2.54±

0.52

0.10±

0.02

0.68±

0.14

0.74±

0.09

1.59±

0.21

1.92±

0.17

0.13±

0.07

0.98±

0.34

1.60±

0.24

2.14±

0.42

Pirimiphos-methyl

2.57 ±

0.14

0.16 ±

0.03

0.43 ±

0.06

1.85 ±

0.11

2.42 ±

0.36

4.44±

0.61

0.27±

0.05

0.78±

0.19

3.62±

0.32

4.36±

0.15

5.89±

0.51

0.66±

0.06

2.10±

0.35

9.63±

0.87

10.62±

0.84

Profenofos 4.46±

0.33

0.37±

0.05

1.59±

0.21

2.55±

0.34

3.86±

0.36

7.36±

0.35

0.22±

0.08

2.26±

0.26

4.05±

0.65

6.61±

0.45

8.42±

0.74

0.69±

0.07

3.91±

0.30

8.83±

0.34

12.62±

0.75

Control 4.70±

0.44

4.47±

0.44

5.25±

0.24

6.33±

0.36

7.30±

0.19

11.42±

1.02

13.34±

1.24

15.44±

2.41

14.91±

2.21

21.09±

2.07

20.30±

0.82

33.62±

3.48

41.91±

3.41

61.71±

2.89

62.80±

2.78

the second spray (population density just before sprying) ,** the third spray (population density just before sprying). S.D: Standard deviation.

159


Table ( 20 ): Efficiency of certain pesticides against the cotton aphid, Aphis gossypii in squash plants.



0.17 e

99.35±

0.22 c

99.47±

0.09 c

95.48±

0.63 c

81.59±

0.03 c

99.23±

0.27 b

98.83±

0.19 d

96.69±

0.50 d

95.08±

1.18 d

85.106±

2.70 c

93.15±

0.21 b

80.98±

2.39 ab

61.37±

7.81 dc

41.51±

17.85 ab

87.69

Pirimicarb 500 gm. 96.45 ±

0.19 d

87.02±

1.08 b

75.03±

7.54 b

74.59±

3.99 b

69.64±

6.86 bc

98.24±

0.60 b

88.04±

0.24 bc

84.14±

3.80 cd

79.65±

2.23 c

60.21±

9.49 d

92.77±

8.59 b

85.23±

2.81 b

87.05±

2.40 cd

81.95±

3.79 e

82.86


1.27 a

67.76±

7.14 a

57.72±

3.24 a

38.71±

4.60a

30.62 ±

3.36 a

72.16±

11.05 a

75.46±

8.82 a

84.14±

3.79 a

49.84±

2.36 a

21.57±

12.04 a

67.597±

3.73 a

75.68±

2.22 a

48.67±

7.78 a

26.90±

8.19 a

56.42

Chlorpyrifos-methyl

1 L. 99.44±

0.19 e

97.75±

0.78 c

88.87±

0.65 bc

87.56±

0.82 c

75.32±

7.25

99.34±

0.61 b

99.30±

0.32 d

86.45±

4.62 cd

86.84±

6.69 cd

78.94±

8.12 c

99.41±

0.74 b

98.65±

1.138 c

88.90±

7.57 b

89.09±

6.75 c

84.70


1.37 c

85.46 ±

2.32 b

61.83±

16.01a

59.22±

9.06 b

42.54±

9.75 a

99.02±

0.15 b

91.03±

1.20 cd

38.99±

9.98 d

63.86±

4.72 b

51.83±

2.85 b

97.9b ±

1.10 b

89.38±

5.57 b

62.09±

8.25 bc

61.38±

7.05 d

71.32


1.99cd

73.7±

8.07 a

77.31±

7.31 b

65.66±

9.58 b

60.41±

2.52 b

96.3±

1.39 b

79.05±

8.60 ab

77.34±

1.88 c

65.18±

8.19 b

56.72±

5.48 b

95.89±

2.31 b

74.75±

10.8 ba

72.76±

2.64 c

63.58±

9.97 b

75.24

Pirimiphos-methyl

1.5 L. 93.47 ±

0.91c

85.08±

2.74 b

46.54±

2.16 a

39.2±

11.35a

28.71±

11.79 a

94.8±

1.17 b

86.59±

4.5 abc

36.66±

12.63 a

46.11±

8.49 a

24.04±

14.92 a

93.23±

0.90b

82.61±

3.45 b

45.80±

8.25 a

41.26±

8.84 b

60.29


1.85 b

68.21±

3.24 a

57.78±

2.70 a

43.84±

9.38 a

31.95±

3.13 c

96. 3±

1.39 b

77.24±

3.59 ab

58.1±

4.68 b

51.23±

5.38 a

35.46±

8.17 a

95.08±

0.19

77.30±

3.58 ab

65.29±

4.16 c

51.49±

1.30 b

64.32

L.S.D 0.95 2.1026 7.2736 0.0778 12.6711 11.6330 6.9109 803740 11.788 9.6188 15.4353 5.9882 8.5134 11.3677 15.8949

* the second spray . ** the third spray. S.D: Standard deviation. Means followed by the same letter (s) are not significantly difference (P= 0.95 level)

160


0 1 3 5 7 9 10 12 14 16 18 19 21 23 25

Time (days).

0

10

20

30

40

50

60

70

Av e

rage

num

ber

of i n

sect

s / l

eaf.



Fig. (15):The average number of Aphis gossypii / leaf infesting squash plants.

161


considerable effect against the cotton leaf worm, These results are

important when designing a program of pest control as it is something

precious to control more than one insect at the same time particularly

those belonging to certain group (e.g. sucking pests and lepidopteran

insects).

2- Both chloropyrifos-methyl and pirimicarb were relatively of low

toxicity to the beneficial predator, Paedrus alfierii Table (8). Therefore

the two compounds (i.e. chloropyrifos-methyl and pirimicarb) were

subjected to further evaluation for their toxicities to white rats as a

mammalian test organism aiming to detect the possible hazardous effects

which might reflect the noxious effects on human consuming edible parts

of sprayed plant or handling these insecticides during application.

2-Toxicity of chloropyrifos-methyl and

pirimicarb to white rats.

2.1. Acute toxicity:

The acute toxicity as a single oral dose expressed as LD50 values

and their confidence limits was determined. The method of Weil (1952)

was used. Results recorded in Table (22) show that LD50 values of

chloropyrifos-methyl and pirimicarb are 1200 and 70.7 mg a.i/kg b.w. of

rats, respectively. These values are somewhat in the range of those

reported by Thomson(1983) (LD50: 2140 and 147 mg/kg b.w of rats for

chloropyrifos-methyl and pirimicarb, respectively). The severity of the

acute toxicity symptoms for both compounds was dose-dependent. Sings

of toxicity appeared as tremors, decreased appetite, abdominal cramps,

salivation, diarrhea, back legs paralyzation, convulsions and coma which

was often terminated by death. These symptoms are similar (to some

extent) to those of human intoxication of organophosphate and carbamate

insecticides reported by Kaloyanova and El-Batawi (1991). Death and

162


Table (21): Average % reduction of different insect pests infesting squash and okra

crops as affected by the tested pesticides.

Squash OkraAverage reduction percentage

Pes t i c i des A. gossypii WhiteflyB. tabaci

S. littoralis WhiteflyB. tabaci

Adult stage nymph stage Adult stage nymph stage

Carbosulfan 87.69 36.28 63.26 22.66 77.54 74.28

Pirimicarb 82.86 24.39 49.54 2.72 58.82 30.27

Propoxur 56.42 41.65 28.95 8.30 26.42 36.96

Chlorpyrifos- methyl 84.70 27.62 83.13 81.52 31.49 35.56

Dimethoate 71.32 25.05 57.06 2.07 60.11 43.66

Phenthoate 75.24 60.31 74.88 49.89 58.32 20.57

Pirimiphos-methyl 60.29 80.57 55.86 41.13 64.48 78.92

Profenofos 64.32 43.54 77.34 83.52 70.56 78.62

Table ( 22 ): Acute toxicity of the tested insecticides expressed (as LD50 values and

their confidence limits.) against rats.

Insesticides LD50 (mg/kg b.w) Confidence limits

Chloropyrifos-methyl 1200 600:2400

Pirimicarb 70.7

163


onset of the intoxication symptoms occurred soon after administration of

pirimicarb and after about 6 hours for chloropyrifos-methyl. Kaloyanova

and El-Batawi, (1991) reported that, the interval between exposure and

onset of the symptoms may be as short as a few minutes but is usually 1

to 2 hrs. However, a case of clinically unrecognized protracted poisoning

that started 6 h after the working day of a tractor sprayer was reported

(Devine et al. 1986).

Acute toxicity as described in standard reference books usually

refers to the toxicity of the active ingredient or technical material.

However, for practical purposes, the pesticide user is more interested in

knowing the toxicity of the particular formulation he is using (Oudejans,

1991). On the other hand, pesticides are selective in their action and

different species of animals react differently. So, LD50 values for rats or

beagl dogs may have little bearing on the value for humans. Nevertheless,

LD50 values are very useful in classifying pesticides according to their

toxicity and as a general guide, the probable dose for an adult human

weighing 80kg may be derived from the acute oral LD50 for test animals

(Oudejans, 1991). Thus, LD50 values are not absolute and should be used

only as a guide to relative toxicity. It is wise also to assume that humans

are at least as sensitive to chemicals as the most sensitive animals used in

the tests. According to the tabulation of toxicity rating given by Gosselin

and Hodge (1976), results of the present study revealed that,

chloropyrifos-methyl could be classified as "moderately toxic compound"

and "very toxic" for pirimicarb (see Table 23).

Table(23): Tabulation of acute toxicity classes(Gosselin and Hoge, 1976)

164


Toxicity rating commonly used term LD50 single oral dose(rats)( mg/kg)

6 Super toxic <5

5 Extremely toxic 5-50

4 Very toxic 50-500

3 Moderately toxic 500-5000

2 Slightly toxic 5000-15,000

1 Practically non toxic >15,000

These results are very important for humans consuming edible parts of

vegetables collected directly after spraying especially with pirimicarb or

for those exposing to these compounds during their handling and

application. Thus, pre-harvest periods after applying these insecticides on

vegetables should be greatly considered. Safety precautions during

application must be adopted as well. Also, growing vegetables (e.g. okra

and onion) inside or beside cotton fields, as commonly practiced by

Egyptian farmers, must be avoided where cotton plants are often subject

to massive application of pesticides.

2.2. Sub-chronic toxicities:

Rats were given daily single oral doses each equivalent to 1/10 and

1/30LD50 of chloropyrifos-methyl or pirimicarb. The treatments continued

for 30, 60, or 90 days. Moralities and clinical symptoms of toxicity were

recorded throughout the test period. After each experimental period,

animals were weighed, slaughtered and the tested biochemical parameters

were measured in serum. The effects of the tested insecticides on the

specific activities or levels of ChE, non-specific esterases, enzymes and

biomarkers of liver and kidney functions were determined. The measured

biomarkers of liver function included aspartate aminotransferase, AST

(formerly, glutamate oxaloacetate transaminase, GOT); alanine

aminotransferase, ALT (formerly, glutamate pyruvic transaminase, GPT);

alkaline phosphatase, ALP; total protein; total bilirubin; albumin and

cholesterol while those of kidney function were creatinine and uric acid.

165


Tissues of kidneys and livers were subjected to histopathological

examination.

2.2.1 Clinical symptoms and moralities through test period:

Rats were daily examined physically and clinically through the

duration of the study. The observations included changes in fur textures,

skin, eyes, mucus membranes, orifices and clinical signs of respiratory

behavior changes and others. Special attention was given to examine for

any palpable mass which may be related to tumur incidence. Moralities

occurred through test period are presented in Table (24) from which it is

obvious that, rats given daily single oral doses of 1/10LD50 chloropyrifos-

methyl for 30, 60, or 90 days showed moralities of 14, 29 or 29%,

respectively through the test period. Chloropyrifos-methyl at 1/30LD50

and pirimicarb at the two tested doses showed no moralities. In

chloropyrifos-methyl treated rats, death was onset after about six weeks.

Before death rats showed severe bulging and bloody eyes, back leg

paralyzation, lack of appetite, diarrhea, raising the tails and difficulty in

breathing. The same symptoms were observed for survival of

chloropyrifos-methyl treated rats or rats given daily oral dose 1/10LD50 of

pirimicarb. In the case of daily oral dose, 1/30LD50, the clinical symptoms

were less powerful.

Table (24): The mortality percentage of rats treated with chloropyrifos methyl and

pirimicarb as daily oral doses.

166


Pesticides doses % mortality

Treatment period (days).30 60 90

Chloropyrifos-methyl 1/10 LD50 14 29 29

1/30 LD50 0 0 0

Pirimicarb 1/10 LD50 0 0 0

1/30 LD50 0 0 0

2.2.2. Effects on ChE and esterases specific activities:

The effect of the OP insecticide, chlorpyrifos-methyl as well as the

carbamate, pirimicarb on serum ChE at the two tested daily doses(i.e.

1/10 and 1/30LD50) was investigated. Results are presented in Table (25)

and Fig (16) revealed that ChE activity in rats treated at each of the tested

doses of chloropyrifos-methyl or pirimicarb was significantly depressed.

Chloropyrifos methyl was highly more potent in this respect. Maximum

inhibition was obtained after daily treatment of 1/10LD50 for 90 days

(activity as % of control: 23.9). Pirimicarb was comparatively of low

inhibitory effect. Maximum inhibition was produced after daily treatment

with 1/10LD50 for 60 days (% of control: 66.77). The low inhibitory effect

of pirimicarb against ChE activity might be explained on the basis that,

inhibition of ChE by carbamates is "reversible", while the OP compounds

are often "irreversible" inhibitions since the hydrolysis of phophorylated

enzyme is very low (Wallace and Herzberg, 1988). It is evident that,

although primicarb showed high acute toxicity compared with

chloropyrifos methyl, it proved to be of lower inhibitory effect on AChE.

In general, the results are in agreement with those observed by

Salem et al. (1979); Abbassy et al. (1980 & 1981); Kobayashi et al.

(1986); Sheremet (1986); Fayez and Kilgora (1992); Fossi et al.(1992);

Holmes and Sundaram (1992); Pope and Chakraborti (1992);

Shalaby et al. (1995) and Soliman et al. (1995). They all concluded that

167


AChE activity was significantly decreased after exposure to OP or

carbamate insecticides in many kinds of animals. Inhibition of whole

blood ChE activity by 30% of the pre-exposure was proposed as a hazard

level (Kaloyanova, 1959). Hence, our results suggested that repeated

exposure to chloropyrifos-methyl might be of adverse effect in this

respect.

For esterases results in Table 26 and Fig (17) showed that, the

activity was highly inhibited in rats dosed at 1/10 or 1/30 LD50 for 90

days (at dose 1/10LD50, the specific activities were 38.8 and 52.56% of

control, at dose 1/30 LD50, the specific activities were 44.04 and 62.16%

of control for chloropyrifos-methyl and pirimicarb, respectively). The

effect was less powerful in rats treated for 30 or 60 days (activity ranged

67.7-89.54% of control). Esterases also were found to be affected by

many of anticholinesterases (Lapadula et al. 1985; Abbassy, 1988 a, b ;

Lock and Johnson, 1990; Fossi et al. 1992).

2.2.3 The effect on liver function:

Before presentation and discussing the results it necessary to

elucidate the basis of assessment of liver function. According to Burtis

and Edward (1994), tests of hepatic function could be classified into

three categories.

(1)- Tests of hepatic synthetic function and this includes the tests that

based on the determination of substances produced or synthesized by the

liver (e.g. albumin, cholinestarase).

(2)-Tests of metabolic function and these based on the determination of

substances metabolized by the liver (e.g. bilirubin, cholesterol,

triglycerides, drugs and xenobiotics).

(3)-Tests of hepatic excretory function and this included those based on

the determination of substances released from damaged tissue (e.g.

168


endogenous compounds released by damaged hepatocyte such as the

transaminases (GOT & GPT) and alkaline phosphatase).

The effect of chloropyrifos-methyl and pirimicarb on alkaline

phosphatase; transaminases, ALT & AST; cholesterol; bilirubin; albumin;

and total protein representing the whole evaluation of liver function .

Results are presented in Tables (27-33) and Figs (18-24).

Alkaline phosphatase activity was significantly elevated in the

rats treated at 1/10 or 1/30 LD50 for all tested periods (Table 27 and Fig

18). Similar effects were obtained by many authors who reported that

there were significant increases in the activities of alkaline phosphatase in

different animals treated with various pesticides( Khalifa et al. (1986),

Khorunova and Baranova (1987), Abbassy et al. (1988), Mohamed et

al.(1988), Ray et al.(1988), Ashour et al. (1989), Chambere and

Chambers (1989) , El-Harrawie et al. (1991), Hanafy et al. (1991),

Kandil et al. (1991), Rajeev et al.(1991), Abdel Baki (1993), Abdel

Baki et al. (1993), Ammar and El-Sheikh (1993), El- Hamady (1993)

and Shalaby et al. (1995). The elevation in serum alkaline phosphatase

activity could be attributed to hepatobiliary affection. This may be due to

increase synthesis and release of the enzyme by the damaged hepatic cells

(Murphy 1966; Enan et al. 1982 and Enan, 1983). On the other hand,

alkaline phosphatase is elevated in certain osteologic diseases. e.g. rickets

(Hayes, 1989). This might explain why some of dosed rats in the present

study were paralyzed and were unable to walk.

For transaminases, results recorded in Tables (28,29) and depicted in

Figs(19, 20) show that GOT and GPT activities exhibited fluctuated

effects between increase and decrease through the experimental period at

each of the tested doses but GOT in general was of significant increase.

For the two doses, after 90 days the activity of GPT returned to the

normal level. This could be explained on the basis that the possible

169


mechanism involved in the elevation of transaminases may be due to

tissue damage (Rouiller,1964 and Korsrud et al. 1972), but if the cells

of tissues are destroyed, the source of enzyme decreases, consequently

enzyme activity decreases (Mandal et al. 1992). Transaminases are

important and critical enzymes in the biological processes. They play a

role in amino acids metabolism and biosynthesis. Consequently they are

considered as specific indicators of liver damage. GPT is more specific

than GOT in this respect (Wilkinson, 1970). Also, Hayes, (1989)

reported that, the activity of hepatic enzymes (e.g. GOT, GPT and ALP)

released into the blood by the damaged liver is one of the most useful

tools in the study of hepato-toxicity. Many investigators had measured the

high transaminases activities as indicators to liver injury (Salem et al.

1979), Abbassy et al. 1988; Abbassy et al. 1989 a & b; Ashour et al.

1989); Reena et al.(1989); Zidan et al. 1989 and Eweis et al. (1995).

Based on the obtained results, elevated ALP, GOT and GPT

activities in chloropyrifos-methyl or pirimicarb-treated rats might be due

to hepatoxic effect of these compounds which appeared as liver

dysfunction or liver impairment. The changes of these enzymes in rats

treated for 30 days might be an early identification of liver injury

170


Table (25): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on the specific activity serum ChE in male white rats.

Activities (m mole AsCI/min./mg protein) of periods:-90 day60 day30 day

1/30LD501/10LD501/30LD501/10LD501/30LD501/10LD50I n s e c t i c i d e s% cont.Mean ± sd

10-4% cont.Mean ± sd





10-436.951.36 ±0.05a23.900.88 ± 0.08a50.191.92 ± 0.22a34.331.32 ± 0.29a52.632.10 ± 0.07a36.961.47 ± 0.12 aChloropyrifos-methyl81.303 .00 ± 0.07b72.412.67 ± 0.31b82.643.17 ± 0.40b66.772.56 ± 0.44b89.783.58 ± 0.10b71.992.87 ± 0.45 bPirimicarb1003.69 ± 0.52b1003.69 ± 0.52c1003.83 ± 0.18c1003.83 ± 0.18c1003.99 ± 0.15 c1003.99 ± 0.15 cControl.

0.600.700.700.640.220.56LSD0.95

Means followed by the same letter (s) are not significantly different (P= 0.95 level)

Table (26): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serum esterases specific activity in male white rats.

Activities (m mole a- naphthol/min./mg protein) after periods:-90 day60 day30 day

1/30LD501/10LD501/30LD501/10LD501/30LD501/10LD50I n s e c t i c i d e s% cont.Mean ± sd






10-644.043.70 ± 0.09a38.832.93 ± 0.11a75.445.48 ±

0.062a67.734.92 ± 7.62 a84.516.07 ± 0.10 a77.795.59 ± 0.27 aChloropyrifos-methyl

62.164.69 ± 0.55 b52.583.96 ± 0.20b78.145.68 ± 0.18a77.375.62 ± 0.11a89.546.43 ± 0.36 a80.095.75 ± 0.15bPirimicarb1007.54 ± 0.43 c1007.54 ± 0.43c1007.27 ± 0.06 b1007.27 ± 0.06c1007.18 ± 0.13b1007.18 ± 0.13 cControl.

0.810.570.260.180.450.38LSD0.95


171


30 60 90

Time / days.

0

20

40

60

80

100

% c

ontr

o l.

1/10LD50 chloropyrifos methyl. 1/10LD50 pirimicarb.

1/30LD50 chloropyrifos methyl. 1/30 LD50 pirimicarb.

Fig.( 16 ):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum ChE activity of rats.

172


30 60 90

Time / days.

20

40

60

80

100

% c

ontr

o l.



Fig.(17):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum esterases activity of rats.

173


Table (27): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serum alkaline phosphatase activity in male white rats.

Activities (U/L.) after periods:-

90 day60 day30 dayI n s e c t i c i d e s1/30LD501/10LD501/30LD501/10LD501/30LD501/10LD50

% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd142.0196.00 ± 11.29b219.03148.07± 31.40b122.1380.93 ± 4.83b209.29138.67 ± 6.50b106.7371.93 ± 3.81b221.17149.07 ± 2.44cChloropyrifos-methyl

143.1096.73 ± 9.50b106.1171.73 ± 10.07a126.2683.67 ± 5.13b204.23135.33 ± 2.20b110.5874.53 ± 1.94a180.42121.60 ± 6.50bPirimicarb

10067.60 ± 7.21a10067.60 ± 7.21a10066.27 ± 8.82a10066.27 ± 8.82a10067.40 ± 2.12a10067.40 ± 2.12aControl.

10.990138.937313.021612.89925.50148.3763LSD0.95


174


30 60 90

Time / days.

0

50

100

150

200

250

% c

ontr

o l.



Fig.(18):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum alkaline phosphatase activity of rats.

175


Table (28): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serum ALT(GPT) activity of male white rats.

Activities (m/ml.) after periods:-90 day60 day30 day

1/30LD501/10LD501/30LD501/10LD501/30 LD501/10LD50Insecticides% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd

85.2632.22 ± 1.60b61.4023.20 ± 2.38b79.3730.34 ± 1.41a121.1346.31 ± 3.97b71.2925.65 ± 4.17a64.5023.20 ± 2.18bChloropyrifos-methyl53.6120.26 ± 0.99b35.2313.31 ± 3.34a133.5751.06 ± 4.55b148.6456.82 ± 1.15c59.9721.58 ± 1.31a2910.93 ± 1.42aPirimicarb10037.79 ± 4.94b10037.79 ± 4.94c10038.23 ± 5.17a10038.23 ± 5.17a10035.98 ± 3.90b10035.98 ± 3.90cControl.

6.09937.40578.108017.62906.75345.40297LSD0.95


Table (29): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serum AST(GOT) activity of male white rats.

Activities (m/ml.) after periods:-90 day60 day30 day

1/30LD501/10LD501/30LD501/10LD501/30LD501/10LD50Insecticides% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd% cont.Mean ± sd

132.0998 ± 1.56c129.5896.78 ± 4.34b122.5994.06 ± 4.57c142.75109.52 ± 4.63c

115.07111.73 ± 2.70c

154.69122.10 ± 8.38c

Chloropyrifos-methyl109.3381.65 ± 1.02b139.14103.91 ± 2.5774.0856.84 ± 2.62a82.2863.13 ± 2.52c111.5690.83 ± 3.68b137.25108.33 ±

5.11bPirimicarb

10074.68 ± 3.39a10074.68 ± 3.39a10076.72 ± 0.29b10076.72 ± 0.29b10078.93 ± 1.47a10078.93 ± 1.47aControl.4.46587.00816.08346.08385.527111.4495LSD0.95


176


30 60 90

Time / days.

0

20

40

60

80

100

120

140

160

% c

ontr

o l.



Fig.(19):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum ALT activity of rats.

177


30 60 90

Time / days.

0

20

40

60

80

100

120

140

160

% c

ontr

o l.



Fig.(20):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum AST activity of rats.

178


produced by the tested insecticides, These findings are ensured by other

biochemical and histopathological studies mentioned below.

For cholesterol, results presented in Table (30) and Fig (21) show

that, chloropyrifos-methyl has no significant effect on cholesterol level at

the two tested doses when administration for 30 or 60 days but the level

was significantly increased when the duration of treatment was 90 days.

Pirimicarb showed significant alterations in the level of cholesterol for all

tested periods (the level was decreased in the period 30 or 60 then

increased for the period 90 days). Elevation of cholesterol in tissues of

animals treated with several kinds of pesticides at various doses was

frequently reported (Raj et al.1988; Reena et al. 1989; Ali and Shakoori

1990; Ghosh 1990; Kadyrova et al. 1990; Ogata and Izushi 1990;

Fujitani et al. 1993 and Katayama 1993). In contrast, Matin et al.

(1990) found that cholesterol content of adrenals was depleted in

diazinon-treated animal. Also, Shakoori et al. (1992) found that

cholesterol level was decreased 40% and 66% after 15 and 30 days of

treating rabbits with cyhalothrin. Cholesterol is the initial starting point in

many metabolic pathways. Although a portion of the body’s cholesterol is

derived from dietary intake, most tissue and plasma cholesterol is

synthesized endogenously by the liver and other tissues from simpler

molecules (Burtis and Edward, 1994). Total cholesterol increase could

be considered as due to liver disease (Coles, 1974).

Results of the effect of repeated dosing of chloropyrifos-methyl

and primicarb on serum total bilirubin are recorded in Table(31) and

depicted in Fig(22). Results showed that, the insecticides had fluctuated

effect on bilirubin concentration depending on dose, duration of treatment

and the kind of the tested compounds. At duration 30 or 60 days,

chloropyrifos-methyl caused significant increase in the concentration of

bilirubin whereas pirimicarb led to the same effect after 30 days only. For

179


the duration, 90 day both insecticides significantly reduced the

concentration. The results are somewhat in parallel to those obtained by

many authors. Ali and Shakoori (1990) found that bilirubin content was

increased after 15 days in rats fed aldrin-mixed diet at doses of 2 mg and

2.5 mg/kg body wt./day. Rats fed barley grain ( treated with meothrin at

20 and 40 p.p.m. and storage for 60 days) for 15 days showed increase in

serum bilirubin (Mohamed et al. 1988). In contrast, Shakoori et al.

(1990) found that bilirubin was decreased 35% in rabbits treated with 6

mg/kg body weight of bifenthrin for 30 day.

To explain the important role of bilirubin in metabolic function of

liver, Burtis and Edward (1994) reported that bilirubin, an orange-

yellow pile pigment, is produced mainly from haemoglobin that released

from senescent erythrocytes and destroyed in the reticuloendothelial cells

of the liver, spleen and bone marrow producing bilirubin. After

production, in the peripheral tissues, bilirubin is transported to the liver in

association with albumin. Bilirubin is then rapidly taken up by

hepatocytes. Inside the hepatocytes, bilirubin is rapidly conjugated with

glucuronic acid to produce bilirubin mono- and diglucuronide, which are

then excreted into bile. Once secreted into intestinal tract, bilirubin

glucuronides are hydrolyzed to the unconjugated pigment. The

unconjugated bilirubin is then reduced by the anaerobic intestinal

microbial flora to a group of tetrapyrrol compounds called urobilinogens.

Up to 20% of the urobilinogens are reabsorbed from the intestine and

enter the enterohepatic circulation. Two types of disorders result from

disturbances of bilirubin metabolism (i.e. disturbances due to effect on

production, uptake, storage, metabolism and excretion of bilirubin). The

first type is referred to as unconjugated hyperbilirubinemia jaundice

where serum unconjugated bilirubin concentrations are greater than

normal. This often is due to incomplete maturation of several steps

180


involved in bilirubin metabolism and excretion (e.g. the conjugation

process may be suppressed by toxins). The second type is called

conjugated hyperbilirubinemia (obstructive jaundice) and results from

blockage to the outflow of bile in the biliary tract. As a result, plasma

concentration of conjugated bilirubin increase to abnormal. Generally, in

terms of detecting and quantifying hepatic damage, bilirubin is of

particular interest (Hayes, 1989). Based on the earlier discussion, it is

evident that the raised amounts of serum total bilirubin may indicate to

the alteration in the capacity of the liver to conjugate bilirubin referring to

liver dysfuction.

Results concerning the effect on total protein are presented in

Table (32) and Fig(23). Apart from the dose 1/30LD50 at duration, 30 day,

total protein was significantly decreased in rats treated with

chloropyrifos-methyl or pirimicarb for 30 or 60 days. When rats were

treated for 90 days, significant increase was obtained. Many authors

reported that, total serum protein concentrations were increased after

exposure to each of different pesticides (El-Harrawie et al. 1986; El-

Sheakh et al. 1990; Zidan et al. 1991b and Fujitani et al. 1993). On the

other side, many investigators reported that total serum protein was

observed to be decreased after exposure to different pesticides (Saleh et

al. 1986; Mohamed et al. 1988; Raj et al. 1988; Rao, 1989; Shakoori et

al. 1990 and Saleh 1990). Generally alteration in serum proteins

confirms the influence of pesticides on liver function since it is the

primary site for the plasma protein synthesis. Qualitative and quantitative

disturbance of protein synthesis is a consequence of impaired hepatic

181


Table (30): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serum Cholesterol concentration of male white

rats.

Concentration (mg/100 ml.) after periods:-90 day60 day30 day


308.25171.10±9.87c323.71179.69 ± 5.17c

106.2865.38 ± 4.39b104.6064.35±14.01b92.7555.65 ± 3.14b109.1865.51 ± 2.19bChloropyrifos-methyl276.29153.36±8.25b244.85135.91 ±

3.87a72.3844.53 ± 5.69a64.4439.64 ± 3.65a27.0516.23 ± 2.19a47.3428.41 ± 6.53aPirimicarb

10055.51 ± 1.79a10055.51 ± 1.79a10061.52 ± 2.71b10061.52 ± 2.71b10060.00 ± 1.74b10060.00 ± 1.74bControl.14.98637.73058.864516.99414.84758.1886LSD0.95


Table (31): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serum Bilirubin concentration of male white rats.

Concentration (mg/100 ml.) after periods:-90 day60 day30 day


79.390.47 ± 0.02b94.550.56 ± 0.01b111.630.69 ± 0.01c133.720.83 ± 0.14c167.631.04 ± 0.12a247.981.54 ± 0.03cChloropyrifos-methyl60.000.36 ± 0.03a47.270.28 ± 0.01a85.470.53 ± 0.03a59.880.37 ± 0.02a167.631.04 ± 0.03b187.281.17 ± 0.13bPirimicarb1000.59 ± 0.01c1000.59 ± 0.01c1000.62 ± 0.02b1000.62 ± 0.02b1000.62 ± 0.14a1000.62±0.14aControl.

0.05200.01990.03900.16540.21640.2221LSD0.95


182


30 60 90

Time / days.

0

50

100

150

200

250

300

350

% c

ontr

o l.



Fig.(21):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum cholisterol concentration of rats.

183


03 06 09

.syad / emiT

0

05

001

051

002

052

003

.l ort

n oc

%

.lyhtem sofiryporolhc 05DL01/1 .bracimirip 05DL01/1

.lyhtem sofiryporolhc 05DL03/1 .bracimirip 05DL 03/1

Fig.(22):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum bilirubin concentration of rats.

184


Table (32): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serum Total protein concentration of male white rats.

Concentration (g/100 ml.) after periods:-90 day60 day30 day


141.257.20 ± 0.39b163.238.33 ± 0.11c96.634.99 ± 0.59a91.144.71 ± 0.04a100.685.07 ± 0.24a86.594.36 ± 0.05aChloropyrifos-methyl139.217.10 ± 0.52b135.826.93 ± 0.07b104.125.38 ± 0.30b118.736.14 ± 0.11c98.104.94 ± 0.18a88.754.47 ± 0.05bPirimicarb

1005.10 ± 0.20a1005.10 ± 0.20a1005.17 ± 0.24a1005.17 ± 0.24b1005.04 ± 0.04a1005.04 ± 0.04cControl.0.78530.27600.80930.30150.34990.0999LSD0.95

Means followed by the same letter (s) are not significantly difference (P= 0.95 level)

Table ( 33 ): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serum Albumin concentration of male white rats.

Concentration (g/100 ml.) after periods:-90 day60 day30 day


119.614.16 ± 0.20b129.085.23 ± 0.84a101.305.19 ± 0.15b143.146.00 ± 0.01b43.36%2.61 ± 0.30a55.552.45 ± 0.30aChloropyrifos-methyl102.624.84 ± 0.18b118.514.80 ± 0.72a123.914.25 ± 0.30a131.945.53 ± 0035a59.22%1.91 ± 0.29a54.59%2.40 ± 0.54aPirimicarb

1004.05 ± 0.24a1004.05 ± 0.24a1004.19 ± 0.40a1004.19 ± 0.40a1004.40 ± 0.64b1004.40 ± 0.64bControl.0.41241.30280.60820.61630.87861.0268LSD0.95


185


03 06 09

.syad / emiT

06

08

001

021

041

061

081

002

.l ort

n oc

%



Fig.(23):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum total protein concentration of rats.

186


30 60 90

Time / days.

0

20

40

60

80

100

120

140

160

% c

ontr

o l.



Fig.(24):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on albumin concentration on serum of rats.

187


Table ( 34): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serum Creatinine concentration of male white rats.

Concentration(mg/100 ml.) after period:-30 day 60 day 90 day

Insecticides 1/10LD50 1/30LD50 1/10LD50 1/30LD50 1/10LD50 1/30LD50

Mean ± sd % cont. Mean ± sd % cont. Mean ± sd % cont. Mean ± sd % cont. Mean ± sd % cont. Mean ± sd % cont.

Chloropyrifos-methyl 1.14 ± 0.07a 101.09 1.25 ± 0.03a 111.41 1.20 ± 0.02b 111.5 1.13 ± 0.03a 98.27 1.07 ± 0.08a 96.39 1.13 ± 0.21a 91.62

Pirimicarb 1.22 ± 0.01a 108.70 1.20 ± 0.05a 106.52 1.51 ± 0.06c 131.21 1.35 ± 0.09b 117.34 1.99 ± 0.23b 160.73 1.50 ± 0.06b 120.94

Control. 1.13 ± 0.09a 100 1.13 ± 0.09a 100 1.15 ± 0.06a 100 1.15 ± 0.06a 100 1.24 ± 0.04a 100 1.24 ± 0.04a 100

LSD0.95 0.1335 0.12618 0.10571 0.13050 0.27819 0.24971


Table (35 ): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on serumUric acid concentration of male white rats.

Concentration(mg/100 ml.) after period:-30 day 60 day 90 day

Insecticides 1/10LD50 1/30LD50 1/10LD50 1/30LD50 1/10LD50 1/30LD50

Mean ± sd % cont. Mean ± sd % cont. Mean ± sd % cont. Mean ± sd % cont. Mean ± sd % cont. Mean ± sd % cont.

Chloropyrifos-methyl 4.89 ± 0.27b 142.71 3.93 ± 0.22a 114.58 6.60 ± 1.03b 201.30 4.34 ± 0.26b 132.47 5.95 ± 0.35c 160 4.70 ± 0.35c 126.25

Pirimicarb 6.61 ± 0.82c 192.71 4.54 ± 0.71a 132.29 5.57 ± 0.96b 170.13 3.62 ± 0.15a 110.39 5.02 ± 0.28b 135 4.23 ± 0.21b 113.75

Control. 3.43 ± 0.21a 100 3.43 ± 0.21a 100 3.28 ± 0.45a 100 3.28 ± 0.45a 100 3.72 ± 0.21a 100 3.72 ± 0.21a 100

LSD0.95 1.02452 0.895394 1.70145 0.16116 0.57473 0.27970


188


03 06 09

.syad / emiT

06

08

001

021

041

061

081

.l ort

n oc

%



Fig.(25):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum creatinne concentration of rats.

189


30 60 90

Time / days.

50

100

150

200

250

% c

ontr

o l.



Fig.(26):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on serum uric acid concentration of rats.

190


function (Celia and Wilkinson, 1973) while decreased synthesis of proteins

leads to a decrease in the plasma concentration of albumin. (Burtis and

Edward, 1994).

The effect on albumin concentration was studied. Results are presented

in Table(33) and Fig(24). Concentrations were significantly altered (increase or

decrease). The test period, 30 day was characterized by decreasing

concentration whereas the duration 60 and 90 day showed enhanced levels.

After 90 days pirimicarb-treated rats at dose 1/30LD50 showed albumin

concentration within the normal level. Reduction in protein and albumin

concentrations were observed by many authors in animals exposed to various

insecticides (Mohamed et al, 1988; Saleh, 1990; El-hamady 1996). Albumin

is synthesized only in the liver, it is secreted across the sinusoidal surface of the

hepatocyte into plasma. Hypoalbuminemia is a liver disorder thought to be a

consequence of decreased hepatic synthesis of albumin (Burtis and Edward,

1994).

2.2.4 The effect on kidney function:

The effect of chloropyrifos-methyl and pirimicarb on kidney function of

rats treated with daily single oral doses (1/10 or 1/30LD50) for 30, 60 or 90 days

was investigated. Creatinine and uric acid concentrations were measured in

serum of treated rats. Results are recorded in Tables (34 & 35) and depicted in

Fig (25 & 26).

For both insecticides, treatment of the rats with each of the tested doses

for 30 days showed no significant alteration in the concentration of creatinine.

When treatments continued for 60 days, pirimicarb only resulted in significant

elevation of creatinine concentration at the two tested doses (i.e. 1/10 or

1/30LD50). It is evident that, chloropyrifos-methyl showed no significant effect

191


on creatinine concentration especially when given to rats at 1/30LD50 for 30 or

60 days.

For uric acid, administration of each of the two insecticides at dose

1/30LD50 for 30 days caused no significant alteration in uric acid concentration.

At duration, 60 or 90 days, significant increase was obtained for both

insecticides.

Evaluation of blood urea nitrogen and plasma creatinine can be used as an

index of decreased glomerular filtration in the kidney (consequently, the kidney

dysfunction)(Hayes, 1989). Also according to Burtis and Edward (1994),

creatinine clearance is almost universally used for clinical assessment of

glomerular filtration rate and thus used for evaluating renal function. Uric acid

is less indicative in this respect. In humans, uric acid is the major product of the

catabolism of purine nucleotides and its clinical utility of measurement is that it

is a marker of cell turnover and disorders of purine synthesis (Burtis and

Edward, 1994).Thus, results of the present study suggest that chloropyrifos-

methyl and pirimicarb administration to rats at daily single doses(1/10 and

1/30LD50) for 30 days might not be nephrotoxic.

For the duration, 60 or 90 days, pirimicarb might be of renal toxicity

since concentrations of creatinine were significantly elevated. The results are

ensured by the histopathological examination.

Creatine is synthesized in the kidneys, liver and pancreas then transported

in blood to other organs such as muscle and brain where it is phosphorylated to

phosphocreatine, a high-energy compound. Interconversion of phosphocreatine

and creatine is a particular feature of metabolic processes of muscle contraction.

Some of the free creatine in muscle spontaneously converts to creatinine, its

anhydride. Creatinine is released into body fluids at a constant rate and its

plasma levels are maintained within narrow limits and its clearance may be

192


measured as an indicator of glomerular filtration rates (GFR) in kidney. The

greater apparent GFR found by creatinine clearance may be due to an increase

in tubular secretory activity for creatinine when plasma levels increase much

above normal(Burtis and Edward,1994).

2.2.5 The effect on body weight gain:

The effect of the insecticides, chloropyrifos-methyl and pirimicarb on

body weight gain of rats was studied. Rats were treated at the doses 1/10 or

1/30LD50 as a daily single oral doses for 30, 60 or 90 days after which, rats

were weighed and averages of body weight gains were calculated. Results are

recorded in Table (36) and illustrated in Fig(27).

Results revealed that, rats treated with 1/30LD50 of each of the tested

insecticides for 30 or 60 days showed no significant alteration in body weight

gain. However, body weight gain of rats treated with 1/30LD50 of each of the

two insecticides for 90 day were significantly reduced. At a dose, 1/10LD50 all

treatments showed significantly reduction. Many investigators found that

pesticides (differing in their chemical structure) caused marked losses of

weights in various treated animals (Abbassy et al. 1981; El-Harrawie et al.

1986; Lapadual et al. 1985; Raj et al. 1988; Pillai et al. 1989; El-Gendy

1991; Fayez and Kilgore 1992; Fujitoni et al. 1993; Paul et al. 1993; Abdel-

Nasser 1995; Hassan et al.. 1994; Attia 1995 and Eweis et al. 1995.). The loss

of body weight gain might be due to the effect of the pesticides on the

absorption of nutrients by the gastrointestinal tract as suggested by Abou Donia

and Graham(1978). Also, loss of body weight gain might indicate to the

changes in appetite or changes in the efficiency of feed utilization by the body

or generally may related to the hepatic lesions. (El- Fiki et al. 1979).

193


Table (36): Effect of daily oral doses of chloropyrifos-methyl and pirimicarb on body weight gain of male white rats.

30 day 60 day 90 day1/10LD50 1/30LD50 1/10LD50 1/30LD50 1/10LD50 1/30LD50

I n s e c t i c i d e s initial body

weight

End body weight

body weight gain

initial body

weight

End body weight

body weight gain

initial body

weight

End body weight

body weight gain

initial body

weight

End body weight

body weight gain

initial body

weight

End body weight

body weight gain

initial body

weight

End body weight

body weight gain

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Mean±sd

Chloropyrifos-methyl 105.66±

9.60

108.00±

27.05a

1.23±

1.63a

106.33±

3.78

142.67±

12.22ab

34.20±

10.78a

105.67±

1.15

152.33±

2.89ab

44.20±

4.28a

98.67±

7.02

143.67±

12.66a

45.56±

3.89a

106±

7.937

144.67±

3.78a

36.81±

6.78a

99.00±

12.53

147.33±

16.86a

49.03±

5.68a

Pirimicarb 91.66±

0.57

95.00±

4.00a

4.37±

3.25a

89.33±

5.033

119.00±

10.39a

33.84±

18.03a

101.00±

7.21

132.00±

13.00a

31.56±

20.76a

95.00±

5.00

139.33±

4.16a

46.79±

3.79a

101.67±

8.96

163.33±

9.50a

61.01±

7.91a

94.33±

3.21

153.00±

8.88a

62.10±

3.88a

Control. 102.0±

3.60

179.63±

30.72b

76.63±

30.24b

102.0±

3.60

179.67±

30.24b

76.63±

33.72a

100±

1.00

206.00±

45.92b

106.27±

47.93b

100.00±

1.00

206.00±

45.92a

106.27±

47.93a

100.00±

1.00

251.67±

25.97b

151.63±

25.32b

100.00±

1.00

251.67±

25.97b

151.63±

25.32b

LSD0.05 47.02 43.39 39.48 45.82 55.15 60.45 55.15 55.64 32.19 31.58 30.32 37.15


194


30 60 90

Time / days.

0

50

100

150

% c

ontr

o l.

1/10LD50 chloropyrifos methyl. 1/10LD50 pirimicarb. 1/30LD50 chloropyrifos methyl.

1/30 LD50 pirimicarb. control.

Fig.(27):The effect of daily oral dose of chloropyrifos methyl and pirimicarb on body weight gain of rats.

195


2.2.6 Histopathological examination:

Tissues of livers and kidneys of rats treated with 1/10 or 1/30LD50

as a daily single oral doses of chloropyrifos-methyl or pirimicarb were

subjected to histopathological examination.

2.2.6.1 Histopathological effect in livers of treated rats:

Figure(28) shows the normal structure of liver. Microscopical

examination showed that livers of animals treated with chloropyrifos-

methyl at doses 1/10 or 1/30LD50 showed degenerative changes in the

hepatocytes appeared as vascoular degeneration (Fig 29) and albuminous

dystrophy in the centrolobular zone of hepatic lobules. Sinusoidal

dilatation and kuffer cell activation were uncommon. Focal cytic necrosis

was also observed in few animals Fig (30). The abvementioned changes

were observed in livers of most of animals treated for 30, 60 or 90 days

but were more clear in the higher dose and the long period.

For livers of rats treated with pirimicarb at each of the tested doses

and different periods of treatment, the examination showed cloudy

swelling, more prominent vacoular degeneration and sinusoidal dilatation

(Fig 31).

2.2.6.2 Histopathological effect in kidneys of treated rats:

Figure 32 shows cross section in kidney of control rats.

Kidneys of rats treated with chloropyrifos-methyl or pirimicarb at doses

1/10 or 1/30LD50 for 30 days showed normal structure of kidney tissues

and cells Fig (33, 34, 35, 36). For rats treated with chloropyrifos-methyl

at dose 1/10 or 1/30LD50 for 90 days, Kindneys showed mild degenerative

changes of the renal tubules. Many of the tubules showed swelling and

granularity of the cytoplasm of the lining epithelium resulting in

narrowing or oblitration of the tubular luminae (Fig 37). Some of the

renal tubules and collecting ducts showed cystic dilatation of the luminae

196


and atrophy of the lining epithelium (Fig. 38). The changes were more

prominent in rats dosed at 1/10LD50.

For rats treated with pirimicarb at doses 1/10 or 1/30LD50 for

periods 60 or 90 days tissues of kidneys showed cloudy swelling renal

tubular epithelium and cystic dilatation of some renal tubules and

collecting duct (Fig, 39 and 40).

Fig.(28):Show the normal structure of liver (control rats), H&E. X400.

197


Fig. (29): Liver of rats treated with chloropyrifos methyl at 1/10LD50 for 90 days, as daily single oral doses.

Shows cloudy swelling and vacuolar degeneration of hepatocytes. (H & E. X400).

Fig. (30): Liver of rats treated with daily single oral dose of 1/10LD50 chloropyrifos methyl for 90 days. The figure shows local areas of lytic necrosis of the

hepatocytes.(H & E. X100).

198


Fig. (31): Liver of rats treated with daily oral doses of 1/10LD50

pirimicarb for 90 days. The figure shows cloudy swelling and vaculolar degeneration of hepatocytes

with dilating of some hepatic sinusiod. (H & E. X400).

Fig.(32): Show the normal structure of kidney tissue and cells (control rats) H & E. X400.

199


Fig. (33): Kidney of rats treated with daily 1/10 LD50 of chloropyrifos-methyl for 30 days. tissue appears

healthy . (H & E. X400).

Fig. (34): Kidney of rats treated with 1/30 LD50 of chloropyrifos methyl for 30 days, This section shows that renal

tissue seems to be healthy .(H & E. X400).

200


Fig. (35): kidney of rats treated with daily oral doses of 1/10LD50 of pirimicarb for 30 days. Show normal

view of kidney histology. (H & E. X400).

Fig. (36): kidney of rats treated with daily oral doses of pirimcarb for 30 days. showing normal view of tissue

and cells comparing with the control one. (H & E. X400).

201


Fig. (37): kidney of rats treated with 1/10LD50 chloropyrifos methyl as daily oral doses for 90 days. This figure shows cloudy swelling of renal tubular epithelium

with obliteration of lubular luminae. (H & E. X 100).

Fig. (38): kidney of rats treated with 1/10LD50 chloropyrifos-methyl for 90 days, as daily single oral doses. The

figure shows albuminus dystrophy and cystic

202


dilatation of some renal tubules and collecting duct. (H & E. X100).

Fig. (39): kidney of rats treated with 1/10LD50 of pirimicarb for 90 days, as daily single oral dose. This section shows cloudy swelling of renal tubular epithelium. (H & E.

X100).

Fig. (40): kidney of rats treated with 1/10LD50 of pirimicarb for 90 days, as daily single oral dose. This section shows dilatation of many of renal tubular luminae. (H & E.

X100).

203

CONCLUSIONCONCLUSION

CONCLUSION

Among eight insecticides, chloropyrifos-methyl and pirimicarb

were found to be the most advantageous ones as protectants against

certain pests commonly attacking vegetable plants. In addition to their

reasonable efficacy against a wide range of pests, they exhibited a low

toxicity to the tested predator. Chloropyrifos-methyl was of a moderate

acute toxicity. However these insecticides may present many adverse

effects to human and animals especially after repeated exposure.

Although the two insecticides might not be nephrotoxic, they showed

severe hepatotoxicity. So, the study demonstrate the great importance of

using non chemical methods for pest control. In general if chloropyrifos-methyl and pirimicarb have

to be involved in I.P.M programs especially on vegetable plants, repeated exposure must be avioded

as possible. In this respect, safety precautions during application, and preharvest intervals should be

greatly considered.

ENGLISHENGLISH SummarySummary

TOXICOLOGICAL STUDIES OF SOME PESTICIDES

IN RELATION TO THEIR SIDE EFFECTS

SUMMARY

The study aimed to evaluate the insecticidal activity of eight

organophosphate and carbamate insecticides against certain pests

commonly attacking vegetable plants. The pests included sucking pests (i.

e. aphids, whitefly, mites) and the cotton leafworm Spodoptera littoralis.

In this respect laboratory and field experiments (at fields grown with

squash and okra) were carried out. In addition, the toxicity these

pesticides to the predator, paederus alfierii in the laboratory was studied.

The tested organophosphorus were chloropyrifos-methyl, dimethoate,

phenthoate, profenofos and pirimiphos-methyl while those of carbamates

were, carbosulfan, primicarb and propoxur. The most efficient

insecticides (i. e. chloropyrifos-methyl and pirimicarb) were further

evaluated for their mammalian toxicity against white albino rats. So,

acute and subchronic studies were conducted. In subchronic studies rats

were orally given the insecticides at doses 1/10 and 1/30 LD50 for each

insecticide. The treatments were performed daily and continued for 30,

60, or 90 days after which, rats were slaughtered and blood serum were

obtained. In blood serum, biochemical markers representing liver and

kidney functions were measured. In addition the activities of ChE and

non-specific esterase were determined. The effect of the tested

insecticides on body weight was also investigated. Livers and kidneys

tissues were subjected to histopathological studies as well.

SUMMARY

For laboratory studies, using slide dipping technique, results

showed that, profenofos had the highest toxicity against Aphis gossypii

followed by dimethoate, phenthoate, chloropyrifos-methyl, carbosulfan

and pirimicarb (LC50`s values were 0.004, 0.01, 0.05, 0.11, 0.125 and

0.55 ppm respectively). Propoxur and pirimiphos-methyl showed very

weak toxicity (LC50`s: 26.4 and 66.78 ppm respectively). For the aphid,

Brevicoryne brassicae, chloropyrifos-methyl was the most toxic

compound followed by profenofos, pirimicarb, propoxur and carbosulfan

( LC50`s: 1.44, 2.40, 2.95, 4.37 and 5.88 ppm, respectively ). Dimethoate,

phenthoate and pirimiphos-methyl were of weak toxicity (LC50`s: 32.46,

18.5 and 8.64 ppm , respectively). The aphid, Aphis cracivora was the

least susceptible to the tested insecticides. The most toxic compound was

profenofos followed by pirimiphos-methyl (LC50`s 5.45 and 8.25 ppm ,

respectively). For the spider mites, Tetranychus sp, the tested insecticides

exhibited weak toxicity. The most toxic compound was profenofos (LC50:

186.54 ppm). Adopting the leaf dipping technique against the cotton leaf

worm Spodoptera littoralis, the insecticidel showed low to very weak or

no toxicity. Chloropyrifos-methyl and profenofos were the most toxic

(LC50`s: 74.53 and 133.35 ppm, respectively). The rest of insecticides

showed slight toxicity (LC50`s> 1000ppm). Pirimicarb, propoxur and

dimethoate were nearly of no toxicity. For the predator Paederus alfierii,

the toxicity of the compounds could be descendingly arranged as follows:

propoxur> phenthoate > dimethoate > carbosulfan> pirimiphos-methyl>

pirimicarb> chloropyrifos-methyl> profenofos Thus, pirimicarb,

chloropyrifos-methyl and profenofos were the least toxic compound to the

predator (toxicity indexes were 2.74, 0.51 and 0.34 respectively).

For field experiments, three sprays with the tested insecticides at the

recommended rates were applied , separated each other with periods each

of 9 days. In the field, aphids Aphis gossypii and whitefly, Bemisia tabaci

206

SUMMARY

were mainly observed and detected on squash plants while the whitefly

and cotton leafworm S. littoralis could be observed on okra plants.

Generally the insecticides showed low effectiveness against the whitefly.

Apart from, pirimiphos-methyl, profenofos, phenthoate and carbosulfan

whose efficiencies to whitefly were relatively moderate, insecticides

tested were of poor efficacy. The efficiency varied depending on the

tested stage (nymphs or adults) and the type of vegetable plants carrying

the insect (i. e. squash and okra). For aphids, A. gossypii, carbosulfan was

the most effective insecticide followed by chloropyrifos-methyl and

pirimicarb (percents of reduction through the experimental period were

87.69, 84.7 and 82.86, respectively). The least effective compounds were

pirimiphos-methyl and propoxur (% reduction: 60.29 and 56.4,

respectively). Against the cotton leafworm, profenofos and chloropyrifos-

methyl had the highest effects (% of reduction: 83.5 and 81.5

respectively).

Based on the above-mentioned results it is evident that, pirimicarb

and chloropyrifos-methyl have many advantages enough to make them

the most promising among the insecticides tested for many reasons.

Firstly chloropyrifos-methyl had the highest toxicity against aphids taking

in consideration the three species of tested aphids (as shown in the

laboratory studies) .Meanwhile, chlorpyrifos-methyl was the most toxic

compound to the cotton leafworm. Pirimicarb was among the insecticides

that have a relatively reasonable toxicity to aphids. In field experiments,

chloropyrifos- methyl and pirimicarb still having a high efficiency against

A. gossypii on squash plants. On squash plants also chloropyrifos-methyl

was of relatively high effect on immature stage of whitefly. This

compound lasted effective up to 9 days after spraying. On okra plants,

chloropyrifos-methyl exhibited a considerable effect against the cotton

207

SUMMARY

leafworm. Secondly, both chloropyrifos-methyl and pirimicarb were

relatively of low toxicity to the beneficial predator, Paederus alfierii.

Thus the two compounds were further evaluated for their acute and

subchronic toxicity against white rats. Acute toxicity studies showed that

chloropyrifos-methyl was relatively of moderate toxicity to rats (LD50:

1200mg/kg) whereas pirimicarb was of high toxicity. Rats daily treated

orally with 1/10LD50 of chloropyrifos-methyl for 3o days showed 16%

mortality throughout the experimental period and those treated for 60 or

90 days showed 33% mortality. In the serum of rats treated with

chlorpyrifos-methyl or pirimicarb at doses 1/10 or 1/30LD50 for all the

tested periods (30, 60 and 90 days), the activities of AChE and esterases

were significantly inhibited. Rats treated with chloropyrifos-methyl and

pirimicarb at doses 1/10 or 1/30 LD50 for all tested period showed

significant alteration in the activities or levels of biochemical

measurements representing liver function (I. e. alkaline phosphatase,

transaminasees, cholesterol, bilirubin, albumin and total protein). This

might indicate to the possible onset of liver impairment. Biomarkers of

kidney function (i.e. uric acid and creatinine) were not significantly

altered especially when rats were treated for 30 days. Also, body weight

gain of treated rats at all tested doses and tested periods were affected.

This is something logical since body weight expresses the general

hygienic and physiological state of the animal.

The histopathological studies ensured the biochemical ones. All the

tested doses caused liver damage appeared as congestion in the central

vein, massive destruction of the hepatic cord, severe degeneration of the

hepatocytes near the central vein, more diffusion of the lymphocytes and

others Tissues of rats treated with 1/30LD50 for 30 days of both

208

SUMMARY

insecticides showed normal kidneys but those treated for 60 days showed

degeneration of renal tissues and other alterations.

In conclusion the study revealed that, although chlorpyrifos methyl

and pirimicarb have proved to be efficient against various insect pests

commonly attacking vegetable plants and less toxic to the tested

beneficial predator Paederus alfierii, they might be of adverse effects to mammals. The

hazards of these insecticides might be ensured to agricultural workers during the application of these

pesticides. The noxious effects could be easily avoided when safety precautions are adopted during

handling and applying these chemicals. So protecting clothes and devices are of great importance in

this respect. In addition preharvest periods after applying these insecticides must be greatly

considered.

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252

ARABICARABIC SUMMARYSUMMARY

الملخص العربى

الفوسفورية المبيدات من ثمانية سمية وتقيم بدراسة الرسالة تتعلق

- - المن البيضاء الذبابة مثل الخضر آفات من عدد ضد والكاربماتية

من لنوع المبيدات هذه سمية وكذا االحمر - العنكبوت القطن ورق دود

اجراء تم الصدد هذا . وفى الرواغه حشرة وهو النافعة المفترسات

والمركبات والباميه الكوسه نباتات على حقلية واخرى معملية دراسات

، البروبكسر ، البريميكارب ، الكاربوسلفان هى دراستها تم التى

، البروفينوفوس ، الفينثويت ، الدايميثويت ، - ميثايل والكلوربيرفوس

المركبات هذه انسب معرفة الى التوصل .. وبعد ميثايل البيرميفوس

المركبات سمية أختبار تم وحقليا معمليا المذكورة االفات مكافحة فى

والسميه الحادة السميه دراسات خالل من البيضاء الفئران ضد الواعده

. المزمنه تحت

أن وجد معمليا الحشرات ضد االبادى النشاط بدراسة يختص وفيما

يليه البروفينفوس مركب هو القطن من ضد سمية المركبات أعلى

فالكاربورسلفان ميثايل يفوس الكلوربير ثم الفينثويت ثم الدايمثويت

LC( للمركبات القاتلة النصفيه الجرعه قيمة بلغت حيث البريميكارب ثم

،0.125 ،0.11 ،0.051 ،0.01 ،0.004 الترتيب على هى)50

ميثايل والبريمفوس للبروبكسر بالنسبه . أما المليون فى جزء0.055

النصفية ( الجرعة المن من النوع هذا ضد ضعيفه سميه أظهر فقد

) . وبالنسبه التوالى ،على المليون فى جزءLC50 26.4 ، 66.78 القاتلة

سميه أعلى ميثايل الكلوروبيرفوس لمركب كان فقد الصلبيات لمن

النصفيه ( الجرعة فالبروبكسر والبريميكارب البروفينفوس مركب يليه

على المليون فى جزء4.73 ،2.95 ،2.40 ،1.44 هى القاتله

حساسية المن أنواع أقل كان فقد البقوليات لمن بالنسبه ) أما التوالى

العربى الملخص

أظهرت فقد االحمر للعنكبوت وبالنسبة الدراسة تحت للمركبات

ورق لدودة بالنسبه . أما له ضعيفة سميه الدراسة تحت المركبات

من وذلك لها سميه أعلى ميثايل الكلوربيرفوس مبيد أظهر فقد القطن

لسمية وبالنسبة الباميه نباتات على والمعملية الحقلية الدراسات خالل

النافعه المفترسات من وهى الرواغه حشرة ضد المختبرة المركبات

< < الفينثويت : البروبكسر كاالتى السميه هذه ترتيب أمكن فقد

< < البريميكارب ميثايل < البيرميفوس < الكاربوسلفان الدايمثويت

البريميكارب مركبات فان وهكذا < البرفينفوس ميثايل الكلوربيرفوس

الحشره هذه ضد سميه المركبات أقل من ميثايل والكلوربيرفوس

والباميه الكوسه نباتات رش تم فقد الحقلية التجارب فى . أما النافعة

. أيام تسعة واألخرى رشه كل بين الدراسه تحت بالمبيدات رشات ثالث

والذبابة القطن من ، من بكل بشده تصاب الكوسه نباتات أن وجد وقد

على البيضاء والذبابة القطن ورق دودة رصد أمكن حين فى البيضاء

المدروسة المبيدات تأثير ضعف لوحظ عامه . وبصفه الباميه نباتات

ميثايل البيرميفوس مبيدات أستثناء مع البيضاء الذبابة على

متوسطه سميه أظهرت التى والكربوسلفان والفينثويت والبروفينفوس

المركبات أعلى أن وجد فلقد الكوسه نباتات على القطن من ضد أما

. والبريميكارب ميثايل الكلوربيرفوس يليه الكاربوسلفان كان تأثيرا

اعلى أن الباميه نباتات على الحقلية الدراسات أكدت الثانية وللمره

والكلوربيرفوس البروفينفوس هو القطن ورق دودة ضد سميه مركب

معظم ان يتضح السابقه التجارب على وبناء عامه . وبصفه ميثايل

السميه أنخفاض ناحيه ومن االبادى النشاط ناحية من المميزات

ولذا والبريميكارب ميثايل الكلوربيرفوس لمركبات هى النافع للمفترس

فئران ضد سميتهما ناحية من تقيميها إعاده تم قد المركبين هذين فان

( تعادل يوميه بجرعات الحيوانات هذه معاملة تم ولقد البيضاء التجارب

2

العربى الملخص

1/10LD50 1/30 LD50 دم سيرم . وفى يوم90 او60 ،30 ) لمدة

حدوث لوحظ المذكورة االزمنه عند الجرعات بهذه المعاملة الفئران

تمثل التى البيوكيمائيه القياسات بعض مستوى فى معنويه تغيرات

، الكولستيرول ، امينيز الترانس ، القلوى الفوسفاتيز مثل الكبد وظيفة

احتمال الى بذلك مشيرا الكلى والبروتين االلبيومين ، البيلوروبين

البيوكيماويه للقياسات بالنسبه . أما الكبد وظائف فى تعطل حدوث

بها يحدث ) فلم الكرياتينين ، البوليك ( حمض الكلى وظيفة تمثل التى

يوم30 لمدة بجرعات عوملت التى الفئران فى خصوصا معنوى تغير

بصفه لوحظ فقد المعاملة الفئران فى الجسم لوزن بالنسبة . أما فقط

الى يشير وهذا معامله الغير بالحيوانات بالمقارنة انخفاض وجود عامه

الدراسات أكدت وقد . هذا والفسيولوجيه العامه الصحيه الحاله سوء

فى لوحظ حيث البيوكيماويه القياسات مدلوالت الهستوباثولوجيه

وجود المستخدمه الجرعات من بأى عوملت التى الحيوانات كبد أنسجة

الكبديه للخاليا وتدمير أحتقان شكل فى تظهر هستولوجيه تغيرات

أما الكبد تلف على يدل مما الهستوباثولوجيه التغيرات من وغيرها

المبيدين كال منLD50 1/30 بـ المعاملة الحيوانات كلى ألنسجة بالنسبة

. هستولوجيه تغيرات أى تظهر فلم الدراسة تحت

مبيدى ومالئمة نجاح من بالرغم أنه الى الدراسة تشير عامه وبصفة

الماصه الثاقبة الحشرات لمكافحة والبريميكارب ميثايل الكلوربيرفوس

توصى . لذا واضحة خطورة المركبين لهذين أن اال القطن ورق ودودة

هذين وتداول تطبيق عند األمان احتياطات اتخاذ بضرورة الدراسة

بين كافية فترات وجود أهمية االعتبار فى االخذ يراعى كذلك المركبين

متبقياتهما خطورة تقل حتى المحصول جمع وبين المبيدين هذين تطبيق

المأكوله. الثمار على

3

اإلشراف لجنة

عشرى على / محمد الدكتور األستاذ ـ1 - الشيخ كفر ـ الزراعة كلية ووكيل المبيدات أستاذ

طنطا جامعة

الباقى عبد السالم عبد / محمد الدكتور األستاذ ـ2طنطا الشيخ- جامعة كفر ـ الزراعة كلية المبيدات أستاذ

الحمضى السيد / شريف الدكتور ـ3 الشيخ كفر ـ الزراعة كلية المساعد المبيدات أستاذ

طنطا جامعة ـ

وتأثيراتها المبيدات لبعض توكسيوكولوجية دراساتالجانبية

من مقدمة رسالةعبدالله العال عبد المنعم عبد صبرى

لدرجة

اآلفات مبيدات الزراعية العلوم فى الماجستيرالشيخ كفر ـ الزراعة كلية ـ الزراعية العلوم بكالوريوس

م1991طنطا جامعة

على والحكم المناقشة لجنة الرسالة

------------------------------ عشرى على محمد/ الدكتور األستاذ ـ1 كلية ووكيل المبيدات أستاذ

كفر ـ طنطا جامعة الزراعةالشيخ

------------------------------ سالمة السيد أحمد/ الدكتور األستاذ ـ2 قسم ورئيس المبيدات أستاذ

كفر ـ الزراعة كلية المبيداتطنطا جامعة الشيخ

------------------------------ عبد مصطفى/ الدكتور األستاذ ـ3 عباسى اللطيف

كلية وعميد المبيدات أستاذ جامعة ـ بدمنهور الزراعة

األسكندرية

------------------------------ الحمضى السيد شريف/ الدكتور ـ4 بكلية المساعد المبيدات أستاذ

جامعة ـ الشيخ كفر الزراعةطنطا

لبعض توكسيوكولوجية دراساتوتأثيراتها المبيدات

الجانبية

عبدالله العال عبد المنعم عبد صبرى

) ( مبيدات الزراعية العلوم بكالوريوسالشيخ كفر ـ الزراعة كلية

م1991 طنطا جامعة

مقدمة رسالة

الزراعية العلوم فى الماجستير درجة على للحصول(المبيدات)

الشيخ كفر ـ الزراعة كليةطنطا جامعة

م1998

MSC SabryAbdallah

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