STATUS AND IMPACT OF COMMERCIAL AQUA DRUGS AND CHEMICALS ON FISH HEALTH AT FARMER LEVEL

A Thesis

By

MD. MOMINUR RAHMAN

Examination Roll No.: 10 Fish Aqua JD-01M

Semester: July-December, 2011

Registration No.: 32613

Session: 2005-2006

MASTER OF SCIENCE (M. S)

IN

AQUACULTURE

DEPARTMENT OF AQUACULTURE

BANGLADESH AGRICULTURAL UNIVERSITY

MYMENSINGH

NOVEMBER, 2011

STATUS AND IMPACT OF COMMERCIAL AQUA DRUGS AND CHEMICALS ON FISH HEALTH AT FARMER LEVEL

A Thesis

By

MD. MOMINUR RAHMAN

Examination Roll No.: 10 Fish Aqua JD-01M

Semester: July-December, 2011

Registration No.: 32613

Session: 2005-2006

Submitted to the

Department of Aquaculture

Bangladesh Agricultural University, Mymensingh

in partial fulfillment of the requirements

for the degree of

MASTER OF SCIENCE (M. S)

IN

AQUACULTURE

NOVEMBER, 2011

STATUS AND IMPACT OF COMMERCIAL AQUA DRUGS AND CHEMICALS ON FISH HEALTH AT FARMER LEVEL

A Thesis

By

MD. MOMINUR RAHMAN Examination Roll No. 10 Fish Aqua. JD-01 M

Semester: July-December, 2011

Registration No: 32613

Session: 2005-2006

Approved as to style and content by:

........................................................... ......................................................... (Prof. Dr Md. Ali Reza Faruk) (Prof. Dr Gias Uddin Ahmed) Supervisor Co-Supervisor

................................................... (Prof. Dr Md. Ali Reza Faruk)

Chairman, Examination Committee And

Head, Department of Aquaculture Bangladesh Agricultural University

Mymensingh

NOVEMBER, 2011

ABSTRACT

The present study was carried out to know the present status of use of commercial aqua

drugs and chemicals and their impact on fish health. Data were collected through

questionnaire interview with fish farmers, drug sellers, personal contact and market

survey from selected area in Mymensing district. Ten categories of drugs and chemicals

were identified that have been using by fish farmers for different purposes. The

categories included antibiotics, disinfectants, gas removal, oxygen supplier, vitamins and

minerals, growth promoter, insect killer, algae killer, predator killer and pH balance. Fish

health management was one of the major areas where aqua farmers used a lot of

antibiotics and disinfectants. Various pharmaceuticals companies were found to provide

these drugs and chemicals with different trade names to meet the farmers demand. The

companies included Fish Tech, Renata, Firstcare, ACI, ACME, Square, Samco, Novartis,

Eon, Opsonin, Al Madina, Rals, Organic, Padma, Nature care, Advance and SK+F etc.

Different types of fish diseases like epizootic ulcerative syndrome or EUS, tail and fin

rot, red and white spot and dropsy in Shing (Heteropneustes fossilis), Koi (Anabas

testudineus), Tilapia (Oreochromis nilotica) and Pangus (Pangasianodon

hypophthalmus) were observed in the study area. Farmers used a lot of drugs and

chemicals single or combined to overcome disease and health problems of their cultured

fishes. Farmer got good recovery of disease after treating with drugs. During the field

observation some problems were identified in using aqua-drugs which include poor

understanding of farmers about the application of drugs, inadequate withdrawal period

and some adverse effect on fish and human health.

ACKNOWLEDGEMENTS All praise to Merciful and Almighty Allah, whose blessings enabled the author to pursue the

research and to submit this thesis for the degree of Master of Science (M.S.) in Aquaculture.

The author expresses his heartfelt gratitude, sincere appreciation, profound indebtedness and

best regards to his honorable Teacher and Supervisor Dr Md. Ali Reza Faruk, Professor and

Head, Department of Aquaculture, Bangladesh Agriculture University (BAU), Mymensingh

for his scholastic guidance, constant untiring supervision, cordial support, endless

encouragement, precious advices on technical matters, constructive criticism, enthusiasm and

helpful discussion throughout the entire period of research work and preparation of this thesis.

The author is highly grateful and indebted to his respected Co-supervisor Dr Gias Uddin

Ahmed, Professor, Department of Aquaculture, BAU, Mymensingh for his kind cooperation,

constructive criticisms and helpful suggestions for the improvement of the research and the

thesis as a whole.

The author would like to express his gratefulness to all the honorable teachers of the Faculty of

Fisheries for their kind co-operation during the period of study.

The author is really grateful to BFRI for funding a project entitled ‘Impact of Aqua Drugs

and Chemicals on Fish Health and Productivity’ which was really very helpful to conduct the

present research.

Special thanks and cordial gratitude is due to Rana vai (former M.S. student), Md.

Shamsuddin Babu (M.S. student) for their kind co-operation, constant support and inspiration

during the whole research period.

The author feels proud to express his sincere appreciation and gratitude to all his fellow

researcher and friends especially Nazmul, Saiful, Shajib, Foysal, Tushar, Irina Mousumi and

Irin for their friendly cooperation, continuous inspiration and encouragements.

The author can never repay the debt of his beloved parents and relatives, especially for their

inspirations, cooperation, constant encouragement and sacrifices for his higher education.

Finally, cordial cooperation, friendly collaboration, fruitful advice and guidance were received

from many other persons from the start till end of this piece of work. The author is immensely

grateful to all of them and regret for his inability for not to mention everyone by name.

The Author November, 2011

CONTENTS CHAPTER TITLE PAGE ABSTRACT iv ACKNOWLEDGEMENTS v-vi LIST OF CONTENTS vii-xii LIST OF TABLES ix LIST OF FIGURES x-xi LIST OF APPENDICES xii

1 INTRODUCTION 1-3 2 REVIEW OF LITERATURE 4-8 3 MATERIALS AND METHODS 9-12

3.1 Study area 9 3.2 Data collection 9 3.3 Questionnaire survey 9 3.4 Fish Health and disease issues 9 3.5 Histological observation 10-12

4 RESULTS 13-28 4.1 Categories of chemicals in aquaculture 13 4.1.1 Antibiotics 13 4.1.2 Disinfectants 14 4.1.3 Gas removal 15 4.1.4 Oxygen supplier 16 4.1.5 Vitamins 17

4.1.6 Growth promoters 18 4.1.7 Enzyme 18 4.1.8 Insect killer 18 4.1.9 Predator killer 19 4.1.10 Algae kiler 19 4.1.11 pH balance 20 4.2 Pharmaceutical company and widely used aqua-

drugs 20-21

4.3 Impact of aqua drugs on fish health and diseases at farmer level

22-25

4.4 Laboratory study for health check 26-28 5 DISCUSSION 29-31 6 SUMMARY AND CONCLUSION 32

REFERENCES 33-36 APPENDICES 37-39

LIST OF TABLES TABLE TITLE PAGE

1 Time schedule in the automatic tissue processor 10

2 Staining procedure followed during the experiment 11

3 Antibiotics used by aquafarmers 13

4 Disinfectants used by aquafarmers 14

5 Gas removal used by aquafarmers 15

6 Oxygen suppliers used by aquafarmers 16

7 Vitamins used by aquafarmers 17

8 Growth promoters used by aquafarmers 18

9 Mostly used commercial aqua-chemicals in terms of fish ponds 21

10 Impact of aqua drugs on fish health and disease recovery in the study area at Fulpur

23

11 Impact of aqua drugs on fish health and disease recovery in the study area at Muktagacha

24

12 Impact of aqua drugs on fish health and disease recovery in the study area at Fulbaria

25

LIST OF FIGURES

FIGURE TITLE PAGE 1 Different antibiotics used by the aquafarmers. a) Doxy-

oxy, b) Renamycin. 14

2 Disinfectants used by the aquafarmers. a) Polgard plus, b) Aquakleen.

15

3 Gas removal used by the aquafarmers. a) Zeolite Gold, b) Aqua-Magic.

16

4 Oxygen supplier used by the aquafarmers. a) Oxy-Gold, b)Oxylife.

17

5 Different vitamins used by the aquafarmers. a) Ossi C, b)Square Aquamix.

17

.6 Growth promoters used by the aquafarmers. a) Charger Gel, b) Aqua Boost.

18

7 Two insect killers used by the aquafarmers. a) Deletix and b) Sumithion.

19

8 Acurte Gold was investigated predator killer. 19 9 Algae killer, Seaweed. 20 10 Section of almost normal skin-muscle of pangus from

Fulpur. 27

11 Section of normal kidney of pangus from Muktagacha. 27 12 Section of normal liver of pangus from Fulpur. 27 13 Section of normal gill of pangus from Fulpur 27 14 Section of Skin and Muscle from Fulpur showing vacuums

(v), necrosis (n) and ruptured dermis ( ). 28

LIST OF FIGURE (Continued)

FIGURE TITLE PAGE 15 Section of affected gill of pangus from Muktagacha

showing cyst (c), heamorrhages (h) and clubbing (cl). 28

16 Section of affected liver of pangus from Fulpur showing necrosis (n), haemorrhages (h) and vacuums (v).

28

17 Section of affected kidney from Muktagacha showing fungal granuloma(fg), haemorrhages (h), pyknosis (p), vacuums (v), necrosis (n) and degenerating kidney tubules (dt).

28

LIST OF APPENDICES

APPENDIX SUBJECT PAGE I Questionnaire used during data collection 37-39

CHAPTER 1

INTRODUCTION

Aquaculture is one of the most important sectors which play a significant role in the

economy of Bangladesh in term of food nutrition, income, employment and foreign

exchange earnings. It is growing more rapidly than all other animal food producing

sectors. Aquaculture in Bangladesh is also under heavy expansion currently and heading

towards commercialization. Through invention and adaptation of new techniques and

evolve of new culturable species this sector have got a dramatic diversification. However,

different kinds of fish disease are the main obstacle of this potential sector and it

considered as one of the major business risk.

To maintain proper health condition of fish, drugs and chemicals play a vital role. There

are several causes of using drugs in aquaculture which include reducing the entrance of

pathogenic organisms in culture area, reducing the multiplication of pathogenic

organisms, reducing the stressful condition on fish health, reducing or protect the

outbreak of disease and treatment of the disease (FDA, 2001).

A variety of chemicals are used in aquaculture for fish health management. Some

common chemicals include sodium chloride, formalin, malachite green, methylene blue,

potassium permanganate, hydrogen per oxide, copper compounds, glutaraldehyde and

trifluralin (Plumb, 1992). Sodium chloride is an old treatment used for a variety of

diseases of fish. It is especially effective chemical when treating some fungal and

parasitic diseases in fish. Formalin is versatile compound used in a variety of ways in

treating fish. Formalin is primarily as an external parasitic on fish and fish eggs as either

flush, prolonged, or indefinite treatment for fungus control. Potassium permanganate

(KMnO4) is one of the widely used chemical in fish health management. It is a strong

oxidizing agent approved for the purpose to teat ponds. Potassium permanganate is good

for treating external protozoa and external bacterial infections (Plumb, 1992).

According to Food and Drug Administration (FDA) law, a drug is any compound which

has a physiological effect on the animal. There are three options for legal use of drugs to

treat animals, including fish. As set by FDA a drug may be used legally if it is either

generally recognized as safe and effective for its intended use, approved by FDA or the

subject of an investigational new animal drug exemption. The relevant authority should

have responsibility for regulating medicated feeds, drugs and other fish health products.

They will approve products, set tolerance levels for drug residues in edible tissues and

determine specifically how drugs are to be used (FDA, 2001).

Concern is also growing over the use and potential misuse of some of aquaculture

chemicals. The amount of information on chemical use in aquaculture and its significance

for human health assurance, environmental protection and sustainable development of the

sector, has been increasing throughout the last two decade (FAO/NACA, 1995; Plumb,

1995).

In our country pesticides are also used in aquaculture of disease treatment, such as

organophosphates, organotin compounds, rotenone and saponin. dichlovos, trichlorfon,

diptarex, melathion, dursban are widely used organophosphate applied to control

ectoparasitic crestacean infections in finfish culture. Due to high neurotoxicity of

organophosphates, potential effects on health of fish farm workers are also health

hazardous chemical (Alderman et al., 1994).

Application of chemical by aquaculturists, either for preventative or treatment purposes, a

certain portion of the applied substance is released directly to the environment. Cravedi et

al., (1987) reported that the vast majority of Oxytetracycline and oxolinic acid provided

is likely to leave the farm as particulate wastes because of feed wastage and poor

digestive absorption of these drugs. Discharge of these contaminated feeds and faeces is

likely to occur continuously at low concentrations but may be greater at certain periods of

the production cycle such as during tank or pond cleaning. Accumulation of solid wastes

and associated chemical residues near the point of discharge is likely.

With the expansion of aquaculture in Bangladesh, there has been increasing trend in

using more chemicals in fish health management. Commonly used chemicals in

Bangladesh aquaculture are lime, rotenone, various forms of inorganic and organic

fertilizers, phostoxin, salt, dipterex, antimicrobials, potassium permanganate, copper

sulphate, formalin, sumithion, melathion etc. (DoF, 2002; Faruk et al., 2005). Most of the

farmers have only limited or no knowledge at all about the appropriate dosages and

method of application. This is due to lack of information regarding the present status and

consequences of aqua-medicines using in aqua-health management.

Considering the above facts, the present study was conducted to attain the following

objectives:

• To know an updated information of commercial aqua drugs and chemicals; and

• To study the impact of use of such chemicals on fish health at farmer level.

CHAPTER 2

REVIEW OF LITERATURE

Boyd (1979) stated that the lowest concentration of potassium permanganate in which the

pink hue remains after 15 min was considered the endpoint for the treatment of

ectoparasites and skin and gill bacterial infections in freshwater fish.

However, Plumb (1979) warned that the level of potassium permanganate may be safe for

fish if exceed the level approximately 2 mg/L of active ingredient.

Kabata (1985) suggested the use of formalin, malachite green, formalin-malachite green

combination and potassium permanganate in controlling the white spot disease caused by

the protozoan lchthyopthirius multififlis. He also found that a 5 to 10 min bath of

potassium permanganate at 100 mg /L was enough to kill sea lice.

Anonymous (1986) observed that early cases of columnaris disease could be successfully

treated with surfactant bath or prolonged immersion in potassium permanganate or

copper sulfate. However, they suggested the use of antibiotic such as oxytetracycline or

nifurpirinol for successful treatment of columnarles diseases.

Limsuwan (1987) determined the acute toxicity of malachite green to silver barb (Puntius

gonionotus), Nile tilapia (Tilapia nilotica), Gunther's walking catfish (Clarias

macrocephalus), carp (Cyprinus carpio) and snakehead fish (Ophicephalus striatus).

Gunther's walking catfish was the most sensitive to malachite green (96-h LC50, 0.066

mg/litre) and Nile tilapia was the most resistant (96-h LC50, 0,425 mg/litre). It was

concluded that malachite green was highly toxic to fish and use of malachite green at 0.

10 - 0. 15 mg/L He also found that a 5 to 10 min bath of potassium permanganate at 100

mg /L was enough to kill sea lice e for therapeutic treatment of fungal infections and

external parasites of fish could be toxic to some species.

According to Rydlo (1989) for controlling the protozoan parasites Costia necatrix,

Chilodonella cyprini, Ichthyophthirius multifilis and Trichodina sp., and the crustaceans

Argulus foliaceus parasitizing Salmo gairdneri or Cyprinus carpio, the suitable

preparations were sodium chloride, potassium permanganate, slaked lime at pH 10,

calcium hypochloride, formaldehyde, malachite green and masoten.

Bhaumik et al. (1991) carried out investigation in the West Bengal on the effect of

epizootic ulcerative syndrome (EUS) and showed that application of lime in ponds gave

68% positive result.

Alderman (1992) stated that malachite green is an organic dye that has been popular as a

parasiticide and fungicide on fish. It is principally used in hatcheries rather than grow-out

systems. Lengthy withdrawal period is essential following application because of

persistent residues.

Baticados and Paclaibare (1992) used formalin and potassium permanganate in treating

the velvet disease caused by Piscinodinium, a protozoan flagellate in aquarium fish.

Plumb (1992) found that potassium permanganate is good for treating external protozoa

and external bacterial infections. He also stated that Sodium chloride is an old treatment

used for a variety of diseases of fish especially some fungal and parasitic diseases

Smith et al. (1994) reported that oxytetracyclin is one of the most widely used

antibacterials in aquaculture worldwide. The vast majority of oxytetracycline supplied in

mediated feed can be found in hatchery effluent at concentrations that account for nearly

all of the drug supplied.

Inglis (1996) reported that anti-bacterial chemotherapy has been applied in aquaculture

for over 50 years, with early attempts to use sulphonamides in the treatment of

furunclosis.

Chowdhury et al. (1996) found that lime and salt treatment of 250 kg/ha respectively

were most effective to suppress the ulcer disease through pond treatment.

Prasad et al. (1996) investigated the effect of five different antibiotics on EUS affected

fish and found that chloramphenicol and pxytetracycline would be effective drugs in

curing the EUS lesion, tetracycline and streptomycin were found to be less effective in

curing the ulcers.

Singh and Singh (1997) obtained seven isolatets of Edwardsiella tarda and showed that

all the isolates were resistant to calistin and gentamicin, but sensitive to ciprofloxacin,

chloramphenicol, nalidixic acid, nitrifurantoin, ofloxacin and streptomycin.

Rahman and Chowdhury (1999) conducted trisls of chemotherapy to the ulcer disease-

affecting catfish as acase study, the best result was obtained by a successive bath in 12%

NaCl suspention and subsequent oral treatment with commercial oxytetracycline at a

dose of 75mg per kg body weight of fish for 5 days.

Sarker (2000) in an experiment to test drug sensitivity of five isolates of A. sobria found

that most of the isolates were fourd sensitive to Oxytetracycline (OT), Oxolinic acid

(OA) and Chloramphenicol (C) but resistance to Erythromycin and Sulphamethoxazole

(SXT).

Brown and Brooks (2002) observed that in Bangladesh about 52% farmers used

potassium permanganate, while 40% used lime, 11% used salt as adisease treatment, such

as disinfectants, banana leaves, fertilizer, alum and water exchange.

Chowdhury et al. (2003) found that antibiotic like renamycin (oxytetracycline) had

positive effect against bacterial infection at adose of 50 mg/kg body wt/day applying for

dayas and 80-90% fish were recovered under laboratory condition.

Sultana (2004) found that commonly used chemicals in aquaculture were lime, salt, urea,

triple super phosphate (TSP) potassium permanganate, vutamins, antibiotics (mainly

oxytetracycline and chlorotetracycline), rotenone, phostoxin, sumithion, melathion and

some hormones.

Faruk et al. (2005) observed that commonly used chemicals in aquaculture are lime, salt,

urea, triple super phosphate (TSP) potassium permanganate, vitamins, antibiotics (mainly

oxytetracycline and chlorotetracycline), rotenone, phostoxin, sumithion, melathion and

some hormones. It is also found that most of the farmers used chemicals and antibiotics

indiscriminately without knowing their mode of action, doses and appropriate procedures

of application.

Yucel et al. (2005) stated that A, hydrophila, A. caviae showed resistance to ampicillin,

cephalothin and trimethoprim but susceptible to ciptofloxacin and ceftriaxone.

Parimal et al. (2006) tested on the sensitivity of Aeromonas hydrophila and concluded

that bacterium was susceptible to gentamicin, ciprofloxacin, chloramphenicol and

oxyteracline but resistant to erythromycin, nitrofurantion and penicillin.

Faruk et al. (2008) in a recent study found a range of chemicals including antibiotics used

in aquaculture for fish health management and disease treatment. Along with commonly

used traditional chemicals, they found a number of new products with various trade

names like JVzeolite, Geotox, Green zeolite, Orgavit aqua, Fish vitaplus, AQ grow-G,

Oxy flow, Oxy max and O2

-marine were the most widely used compounds. Fourteen

branded antibiotics were found with different trade names for disease treatments of

aquatic animal. Major active ingredients of these antibiotics were Oxytetracycline,

Chlorotetracycline, Amoxicillin, Co-trimoxazole, Sulphadiazine and Sulphamethoxazole.

They reported thirty three pharmaceutical companies were seen active for producing and

marketing of these products. Some of these products have been marketed by different

companies from the countries like India, Thailand, Taiwan, Indonesia, Malaysia, Spain

and USA.

CHAPTER 3

MATERIALS AND METHODS

3.1. Study area

Three upazilas like Muktagacha, Fulbaria and Fulpur of Mymensing district were

selected for the present study to know the status of use of commercial aquadrugs by fish

farmer.

3.2. Data collection

Data were collected through the combination of the following survey techniques:

3.3. Questionnaire survey

Fish farmers, hatchery and nursery owner, farm workers and chemical retailers were

questioned individually. A set of preliminary questionnaire based on the objectives of the

study was prepared. Major topic of questionnaire were the name of chemical, active

ingredients, purpose of use, method of application and dose, duration, source, effects on

environment, price, impact on health and productivity etc. In addition data on farming

practices, general farm management, health and disease problems, seasonality, mortality

etc were also gathered. For the interview, simple random sampling methods were

followed during interview.

3.4. Fish Health and disease issues

In addition with the field observation, fish health and disease condition were verified

through clinical and histopathological analysis. Fish were examined clinically through

observing gross signs, abnormalities, lesions and erosions and external parasites.

3.5. Histological observation

For histopathological study, monthly sample was collected from fishes of selected

farmers pond and various organs such as skin, muscle, gill, liver and kidney by a sharp

scalpel and forceps. Skin and muscle were collected from the place between anterior part

of dorsal fin and lateral line and by removing operculum, gills samples were collected.

For liver and kidney, fishes were dissected and then portions of liver and kidney were

collected. All collected samples were fixed in 10% natural buffered formalin. The amount

of fixative was 10 times to bulk of tissue fixed. The sizes of the samples were 1cm3

. The

preserved samples were taken out and trimmed by scalpel. Trimmed samples were placed

separately in perforated plastic holders and covered by perforated steel covers. Labeling

was made with dark pencil on perforated plastic holders. The samples were then arranged

in a steel rack and processed through an automatic tissue processor (SHADON, Citadel

1000) for dehydration, clearing and infiltration. Alcoholic series of higher concentration,

xylene and paraffin (3 series) were used in the processor maintaining at various time

schedules as mention below:

Table 1. Time schedule in the automatic tissue processor Container Chemicals Time (h.)

01 50% Alcohol 1 02 80% Alcohol 2 03 100% Alcohol 2 04 100% Alcohol 2 05 100% Alcohol 2 06 100% Alcohol 2 07 100% Alcohol 2 08 Xylene 2 09 Xylene 1 10 Molten wax 1 11 Molten wax 2 12 Molten wax 2

The samples were then embedded with melted wax, steel mold and perforated plastic

holder. Proper care was taken for the placement and orientation of skin and gill in steel

molds during the embedding. After embedding, the paraffin blocks were placed on table

to become hard. Then the blocks were placed in a deep freeze for half an hour and after

then steel molds were separated from the paraffin blocks. Trimming was done from the

side and surface of the block by scalpel and a microtome machine (Leica JUNG RM

2035). Embedded blocks were then placed in the deep freeze for 30 minutes before final

sectioning. After having sections, the ribbon of sections was placed on a water bath

(Electrothermal, paraffin-section, mounting bath) at 40˚C. A suitable section was selected

and separated from ribbon, which was finally picked up over a glass slide. To fix the

section, the prepared slide was placed on a hot plate (37˚C) for overnight. The sections

were then cleared with xylene, rehydrated with alcoholic series and stained with

haematoxylin and eosin stains proceeding through various chemicals of different

concentrations and time schedules as mentioned below:

Table 2. Staining procedure followed during the experiment

SL. No. Process Solution Time (min) 01 Clearing Xylene 2 02 Xylene 2 03 Rehydration 100% alcohol 2 04 100% alcohol 2 05 95% alcohol 2 06 70% alcohol 2 07 Running tap water 2 08 Stain Haemotoxylene 10 09 Reduce stain Running tap water 2 10 Counter stain Eosin 12 11 Dehydration 70% alcohol 3 dips 12 95% alcohol 3 dips 13 100% alcohol 2 dips 14 100% alcohol 2 dips 15 Clearing Xylene 2

16 Xylene 2

After staining the sections were mounted with Canada balsam and covered by coverslip.

The prepared slides were left on clean platform to hold the cover slips permanently and

then examined under a compound microscope. Photomicrographs of the stained sections

were done by using a photomicroscope. Comparisons of structure and pathology of

organs were made among treatments.

CHAPTER 4

RESULTS

4.1. Categories of chemicals in aquaculture

Mostly ten different categories of commercial aqua-drugs and chemicals were found to

use by aqua farmers in their aquaculture activities. The categories include antibiotics,

disinfectants, gas removal, oxygen supplier, vitamins and minerals, growth promoter,

insect killer, algae killer, predator killer and pH balance.

4.1.1. Antibiotics

About 11 antibiotics with different trade names were found to use by the aqua farmers for

the treatment of fish diseases. Their name, dose and sources are given in Table 3. Mainly

broad spectrum antibiotics were found to use with mixing in fish feed. Two companies

supplied these antibiotics to the farmers.

Table 3. Antibiotics used by aquafarmers

Sl. No. Trade name Dose Source 01 Renamycin 50 mg/kg b.w. Renata 02 Aquamysine 1-1.5 kg/ton Fish Tech 03 Aquamycine 1-2 /kg feed ACI 04 Renaquine 50 mg/kg b.w. Renata 05 Oxy Dox F 1-2 /kg feed ACI 06 Ascamicyne 250-300 /acre SKF 07 Amoxivet 25-45 mg/kg b.w. Techno 08 Oxysestin 1-2 /kg feed Novartis 09 Orgamysin 60 /50 kg feed Organic 10 Doxioxy 1-2 /kg feed Opsonin 11 Amoxifish 3-5 /kg feed Fish tech

Fig. 1. Different antibiotics used by the aquafarmers. a) Doxy-oxy, b) Renamycin.

4.1.2. Disinfectants

About 10 disinfectants from nine different sources were found to use by the farms. Their

trade name, dose and source are mentioned in Table 4. Farmers use chemicals as the

disinfectant to keep their pond free from pollution or microbial pathogen.

Table 4. Disinfectants used by aquafarmers

Sl. No. Trade name Dose Source 01 Polgard plus 500 ml/acre Fish tech 02 Bactisal 350 ml/acre First care 03 Virex 100-200 /33 dec ACI 04 Biogaurd 250 g/33 decimal Sia agro-vet 05 Lenocide 5 ml/dec Nature care 06 Timsen 1st

2 dose: 80 /33 dec

ndEon

dose: 50 /33 dec

07 Emsen 1st dose: 80 /33 dec 2nd dose: 50 /33 dec

Ethical drugs

08 Aqua cleaner plus

1 L/50 decimal for 3-5 ft depth

Fish world

09 Formalin 1-3 ppm Chemical seller 10 Bleaching

powder 60 ppm Chemical seller

a b

Fig. 2. Disinfectants used by the aquafarmers. a) Polgard plus, b) Aquakleen.

4.1.3. Gas removal

To remove organic and inorganic wastes producing gas in ponds, farmers were seen to

use gas removal agent to their culture pond. About 9 gas removal with different trade

name were found to different farm. Their name, dose and sources are given in Table 5.

Table 5. Gas removal used by aquafarmers

Sl. No. Trade name Dose Source 01 Ammonil 100-200 /acre periodically used every month Novartis 02 Aqua magic 05-08 kg/acre Fish tech 03 Miracol lime

mila 100 /dec ACME

04 Geolite gold 200-250 /decimal (repeat 30-40 days interval) Fish tech 05 Gas stop 400-500 /acre Organic 06 Gas check 200 /acre First care 07 Megageo plus 200/decimal ACI 08 Geo tox 20-25 kg/100 dec (repeat 30-40 days interval) Novartis 09 Gasonex plus 200-400 /acre Fish tech

a b

Fig. 3. Gas removal used by the aquafarmers. a) Zeolite Gold, b) Aqua-Magic.

4.1.4. Oxygen supplier

When dissolved oxygen deficiency occur farmers use different kinds of oxygen supplier.

About 5 oxygen suppliers with different trade name were found to use by the farmers in

the study area (Table 6).

Table 6. Oxygen suppliers used by aquafarmers

Sl. Trade name Dose Source

01 Oxy gold 250-500 /acre Fish tech

02 Oxy life 400 /acre/3-6 ft water depth Square

03 Pure oxy 1-2 kg/1000 m Al Madina 2

04 Oxymax 250-500 /acre/meter depth Eon

05 Oxy flow 250-350 /acre Novartis

a b

Fig. 4. Oxygen supplier used by the aquafarmers. a) Oxy-Gold, b)Oxylife.

4.1.5. Vitamins

Four vitamins with different trade names were found to use by the farmers to increase

disease resistance power and sometime to treat diseases of their cultured fish. Their

name, dose and source are given in table 7.

Table 7. Vitamins used by aquafarmers Sl. Trade name Dose Source 01 Grow fast 1 ml/3-4 liter Rals 02 Revit C 1/5-7 kg feed Opsonin 03 Silver mil 1-2 ml/kg feed Sia agro-vet 04 Ossi-C 4-5 /kg feed for 5-7 days Fish tech

Fig. 5. Different vitamins used by the aquafarmers. a) Ossi C, b)Square Aquamix.

a b

a b

4.1.6. Growth promoters

Farmers in the study area named three different types of chemicals used as growth

promoters of their fish. Their name, dose and source are given in Table 8.

Table 8. Growth promoters used by aquafarmers Sl. No. Trade name Dose Source 01 Charger gel 2-4 /kg feed Fish tech 02 Aqua boost 50 /metric ton Novartis 03 Bio-grow 100 ml/33 decimal Sia agro-vet

Fig. 6. Growth promoters used by the aquafarmers. a) Charger Gel, b) Aqua Boost.

4.1.7. Enzyme

Only one type of enzyme was used by the fish farmers in fish feed named Biozyme. They

used it at a dose of 25-50gm/100 kg feed. The pharmaceutical company Fish Tech

provides the enzyme to the farmers.

4.1.8. Insect killer

Sumithion and Deletix are the two insect killer used by the farmers in their pond to kill

different types of harmful insects. The recommended dose of Sumithion was 5-

10ml/day/3ft depth and Deletix was 25-30 ml/acre for 4 feet water depth. Samco and Fish

Tech were the two companies which provide the chemicals to the farmers.

a b

Fig. 7. Two insect killers used by the aquafarmers. a) Deletix and b) Sumithion.

4.1.9. Predator killer

Farmers use rotenone powder to remove predator fish. Rotenone with a brand name

Acurte Gold is provided by a pharmaceutical company Samco was found in the study.

The recommended dose of Acurte Gold was 15 gm/dec/1 ft depth.

Fig. 8. Acurte Gold was investigated predator killer.

4.1.10. Algae kiler

Seaweed was the only type of algae killer used by the fish farmers. They used it at a dose

of 2-4 liter/acre. The pharmaceutical company Fish Tech provides the Seaweed to the

farmers. It is used to remove or control toxic algae and phytoplankton growth.

a b

Fig. 9. Algae killer, Seaweed.

4.1.11. Chemical use as pH balance

Only one type of pH balance was found to use by the fish farmers named bio-pH. They

used it at a dose of 1ml/ L. The pharmaceutical company Biopharma provides the bio-pH

to the farmers. It is used to control pH in water body.

4. 2. Pharmaceutical company and widely used aqua- drugs

Twenty two pharmaceutical companies were seen to provide commercial aqua drugs and

chemicals to fish farms. Major companies were Fish Tech, Renata, Firstcare, ACI,

ACME, Square, Samco, Novartis, Eon, Opsonin, Al Madina, Rals, Organic, Padma,

Nature care, Advance and SK+F. During the survey, 21 drugs retailers have been visited

and around 50 trades named aqua-drugs and chemicals were recorded. Among those, 15

types are widely used by the farmers as given in Table 9.

Table 9. Mostly used commercial aqua-chemicals in terms of fish ponds

Sl. No.

Trade name Active ingredient Purpose of use Source/ Company

01 Renamycin Oxytetracycline As antibiotic Renata 02 Amoxifish Amoxicillin trihydrate As antibiotic

Fish tech

03 Timsen n-alkyl dimethyl benzyl ammonium chloride-40%

As disinfectants Eon

04 Aquamysine Chlorotetracycline As antibiotic Fish tech 05 Ossi-C Oxolinic acid,

bitaglucan, vit-C As vitamins

Fish tech

06 Aquamycine Oxytetracycline hydrochloride

As antibiotic

ACI

07 Virex Per oxy monosulfate, sodium hydrochloroisocyanorate

As disinfectant ACI

08 Aqua kleen Tetra decile trimethyl ammonium bromide, benzal conium chloride, amino nitrogen

As disinfectant Square

09 Geolite gold SiO2, Al2O3, Fe2O3, TiO2

For removing gas , MgO, etc

Fish tech

10 Oxy Dox F Oxytetracycline hydro-chloride, doxycycline

As antibiotic ACI

11 Polgard plus 3-methyl 4 alkyl two chain brominated compound

As disinfectant

Fish tech

12 Charger gel 1-3 D glucan, poly-saccharides, betain, bitaglucans

As growth promoter

Fish tech

13 Seaweed Elemental copper, inert ingredient

As algae killer Fish tech

14 Bactisal Alkyl benzyle dimethyl ammonium chloride

As disinfectant First care

15 Deletix Deltamethrin As argulus killer Fish tech

4.3. Impact of aqua drugs on fish health and diseases at farmer level

Farmers in the study area cultured mostly Thai pangaus, Koi, Tilapia and Shing. In the

Fulpur, Sing was found to succeptable to disease with 90% prevalence as reported by the

farmers followed by Edwardsiellosis in Pangas (80%), red and white spot on Koi (40-

50%) and EUS (20%) and dropsy (10%) in Tilapia (Table 10). However, in Muktagacha,

Koi was the most succeptable species to white spot disease with 50% prevalence

followed by red spot on pangas (40%), EUS (30%) in Tilapia, Sharputi, Rui, Catla and

Mrigal (Table 11). On the other hand, Sing was reported to be the most succeptable

species to disease with 100% prevalence of swollen abdomen followed by

edwardsiellosis in Pangas (40-50%) in the Fulbaria area (Table 12). Shing had 90-100%

mortality within very short period from unknown reason with no obvious clinical sign but

only swollen abdomen.

Farmers were asked about the use and impact of commercial aqua drugs in recovery of

such disease. They reported that after using commercial drugs single or combination of

many drugs, they got good recovery of fish diseases. In Fulpur, farmers used Renamycin,

Ossi-C and Polgard plus and in Fulbaria, they used Poldard plus and Bactisol for the

treatment of Shing (Table 10, 11and 12). EUS affected tilapia was treated with

Renamycin, Polgard plus and Ossi C with 80-95% recovery from disease in Fulpur and

Muktagacha. Edwardsiellosis affected Pangus were treated with Renamycin, Timsen,

Polgard plus and Ossi C in Fulpur and with Geolite and Timsen in Fulbaria having 80%

recovery in both upazillas. Dropsy was seen with Tilapia in Fulpur upazilla where

farmers used Aquamycin and Ossi C as drugs with a result of 95% recovery (Table 10, 11

and 12). In Fulpur and Fulbaria, Zoothamnium and various spots on skin, scales drops in

some parts of koi, where farmers used Renamycin, Aquamycine, Ossi-C and Polgard

plus drugs as treatmens and achieved 70-80% recovery. Sharpunti, Rui, Catla and Mrigal

were also affected by EUS in Muktagacha and the farmers achieved good result by

applying drugs like Renamycin and Ossi C. It was thus observed that aqua drugs played

excellent role in recovery of fish diseases and maintenance of health.

Table 10. Impact of aqua drugs on fish health and disease recovery in the study area at Fulpur

Area Species Diseases

Prevalence (%)

Clinical sign Commercial aquadrugs used Recovery Affected months

Fulpur

Tilapia EUS

(20%)

Red spot on body surface, lesion on body surface

-Renamycin @ 50 mg/kg body weight -Polgard plus @ 500 ml/acre -Ossi-C @ 3 gm/kg feed

95% August

Tilapia Dropsy

(10%)

Swelling of abdomen, accumulation of fluid in abdomen

-Aquamycine @ 1-2 gm/feed -Ossi-C @ 3 gm/kg feed

95% August

Pangus

Edwardsiellosis

(80%)

Exophthalmia, red spot on abdomen and dorsal side

-Renamycin @ 5g/kg feed -Timsen @ 1st

2

dose: 80 gm/33 dec,

nd

-Ossi-C @ 3 dose: 50 gm/33 dec

gm/kg feed

-Polgard plus @ 5 ml/decimal

80%

November-December

Pangus

Edwardsiellosis

(80%)

Exophthalmia, red spot on abdomen and dorsal side

-Renamycin @ 5g/kg feed -Timsen @ 1st

2

dose: 80 gm/33 dec,

nd

-Ossi-C @ 3 dose: 50 gm/33 dec

gm/kg feed

-Polgard plus @ 5 ml/decimal

80%

November-December

Koi (50%) red spot Zoothamnium and red spot on abdomen, scaleless

-Renamycin @ 5g/kg feed -Ossi-C @ 3 gm/kg -Polgard plus @ 5 ml/decimal

70% November-December

Koi (30-40%) white spot

White spot on tail, gill

-Renamycin @ 5g/kg feed -Ossi-C @ 3 gm/kg feed -Polgard plus @ 5 ml/decimal

80% March-April

Table 11. Impact of aqua drugs on fish health and disease recovery in the study area at Muktagacha

Area Species Diseases

Prevalence (%)

Clinical sign Commercial aquadrugs used Recovery Affected months

Muktagacha

Shing, magur

(10%) no symptom

No symptoms, sudden death, slightly red line on body surface

-Aquakleen @ 0.5-1 liter/acre -Gas check 200 gm/acre -Bleaching powder @ 700 gm/32 dec -Renamycin @ 5g/kg feed -Ossi-C @ 3 gm/kg feed -Polgard plus @ 5 ml/decimal -Virex @ 100-200 gm/33 dec -Aquamycine @ 1-2 gm/feed

95% March-April

Tilapia, sharpunti, rui, catla, mrigal

EUS (30%)

Red spot on body surface, lesion on body surface

-Renamycin @ 50 mg/kg body weight -Ossi-C @ 3 gm/kg feed

80%

August

Pangus

(40%) red spot

red spot on operculum, spoilage on body surface

-Renamycin @ 5g/kg feed -Geolite gold @ 200-250 gm/decimal -Aqua kleen @ 0.5-1 liter/acre

80%

November-December

Shing

(40%) Tail rot

Tail rot

-Polgard plus @ 5 ml/decimal -Aqua kleen @ 0.5-1-liter/acre -Geolite gold @ 200-250 gm/decimal

80%

November-December

Affected months

Koi (50%) white spot

White spot on tail, gill

-Aquamycine @ 1-2 gm/feed -Ossi-C @ 3 gm/kg feed -Polgard plus @ 5 ml/decimal

70% November-December

Table 12. Impact of aqua drugs on fish health and disease recovery in the study area at Fulbaria

Area Species Diseases Prevalence (%)

Clinical sign Commercial aquadrugs used

Recovery Affected months

Fulbaria

Shing (100%) swollen abdomen

Swollen abdomen with occasional spots

-Polgard plus @ 5 ml/decimal

-Bactisol @ 350 ml/acre for 3 ft depth

- December-January

Pangus (40%) red spot Red sign on eye, fin and heat; Movement on surface

-Geolite @ 200-250 gm/decimal -Timsen @ 1st

gm/33 dec, 2 dose: 80

nd

gm/33 dec dose: 50

80

Pangus

Edwardsiellosis (50%)

Speedy movement before death, mouth remains in water surface, Exophthalmia, red spot on abdomen and dorsal side

-Renamycin @ 5g/kg

feed

-Ossi-C @ 3 gm/kg feed -Polgard plus @ 5 ml/decimal -Geolite gold @ 200-250 gm/decimal -Gasonex plus @ 200- 400 gm/acre

70% December-January

4.4. Laboratory study for health check

Clinically all the fishes of various regions did not show any remarkable changes.

Histopathology in the control ponds, skin-muscle, liver, kidney and gill of fish had almost

normal structure (Figs. 10, 11, 12 and 13). However, in the chemical treated ponds, the

above mentioned investigated organs of fishes had remarkable pathological changes like

necrosis, hemorrhage, vacuum, melanocytes and partial loss of organs (Figs. 14, 15, 16

and 17).

Fig. 10. Section of almost normal skin-muscle of pangus from Fulpur.

H & E × 150.

Fig.11. Section of normal kidney of pangus from Muktagacha. H & E ×150.

Fig. 12. Section of normal liver of pangus from Fulpur. H & E × 150.

Fig. 13. Section of normal gill of pangus from Fulpur. H & E × 150.

10 11

12 13

Fig. 14. Section of Skin and Muscle from Fulpur showing vacuums (v), necrosis

(n) and ruptured dermis ( ). H & E × 150.

Fig. 15. Section of affected gill of pangus from Muktagacha showing cyst (c),

heamorrhages (h) and clubbing (cl) . H & E × 150.

Fig. 16. Section of affected liver of pangus from Fulpur showing necrosis (n),

haemorrhages (h) and vacuums (v). H & E × 150.

Fig. 17. Section of affected kidney from Muktagacha showing fungal granuloma

(fg), haemorrhages (h), pyknosis (p), vacuums (v), necrosis (n) and

degenerating kidney tubules (dt). H & E × 150.

14

16

15

17

h n

h

n fg

dt

cl

c

h p

v

n

v

v v

CHAPTER 5

DISCUSSION

Aquaculture in Bangladesh is heading towards commercialization and intensification.

Different types drugs and chemicals have become an essential part for successful

aquaculture production. The present investigation was conducted to know the current

status of commercial aqua-drugs and their impact on fish health. Data was collected

through questionnaire interview, market survey and personal contact with retailers and

representatives of pharmaceutical companies at Muktagachs, Fulpur and Fulbaria upazilla

of Mymensingh district.

In the present study, it was found that mostly ten different categories of commercial

aqua-drugs and chemicals were used by farmer for different aquaculture activities like

disease treatment, pond preparation, disinfectants, growth promotion and improve disease

resistance. All these activities were related for better health management of aquatic

animal. At present 150 products of 40 animal health companies were seen to market at

field level. Moreover, these pharmaceuticals companies found to have very attractive

information leaflet to sell their products to the farmer. Farmers of the selected area used

mainly Amoxifish, Renamycin, Timsen ,Aquamysine, Deletix , Seaweed, Ossi-C ,Oxy

gold, Charger gel, Renaquine, Polgard plus, Vectisol, Aqua kleen, Rotenone , Aqua,

boost, Timsen, Sumithione, Doxioxy, Aqua boost and Virex as trade name for disease

treatment. Faruk et al (2008) also found the similar drugs like JVzeolite, Geotox, Green

zeolite, Orgavit aqua, Fish vitaplus, AQ grow-G, Oxy flow, Oxy max and O2

-marine.

However, in the present study we got some new products with various trade names which

include Ossi-C, Oxy gold, charger gel, Polgard plus and Vectisol.

In the study area different types of fish diseases like EUS, tail rot, fin rot, red spot, white

spot and dropsy in different fish species mainly in Shing, Koi, Tilapia and Pangus were

observed. A number of authors also reported the similar conditions in aquaculture of

Bangladesh (DoF, 2002 and Faruk et al., 2004). Most of the farmers of the selected areas

used drugs and chemicals to control these types of disease. It meant that disease problem

was one of the major concerns in aquaculture of the studied areas.

The present study revealed that commercial aqua drugs have some positive impact on fish

health management and disease treatment at farmer’s level. It was observed that farmers

of the selected areas got good results in disease treatments by applying single or

combinations of various aqua-drugs and chemicals. In some cases after use of drugs they

got about 95% recoveries within a short period of time. These results influenced farmer

to use more commercial aqua-drugs in controlling disease. Sometimes they applied drugs

higher than recommended doses to get quick recovery. Normally in rural aquaculture,

farmer used traditional chemicals in health management such as lime, salt, potassium

permanganate, sumithion , melathion, formalin and bleaching powder. But, they did not

get better results than commercial drugs and chemicals. So, the farmer showed less

interest to traditional drugs for disease control. The uncontrolled uses of drugs sometimes

have got negative impact on fish health. Some pathogen evolved drug resistance ability

due to unconscious and repeated use of drugs. It was not possible to control the disease

in such condition. It was also found that in case of diseased Shing, farmers did not get

any recovery. In a certain region of the study area Shing had 90-100% mortalities within

very short period from unknown reason with no obvious clinical sign.

In the present study, it was observed that about 11 antibiotics with different trade names

were used by the farmer. Antibiotics should be used only for the treatment of bacterial

diseases. It was found in the present study that antibiotics were used indiscriminately

without knowing the exact reasons of disease. Some farmers did not follow the described

dosages for treatment. It is widely recognized that the excessive use of antibiotics

contributes the development of resistant strains of bacteria (Inglis, 1996).

In the field level, it was observed that some aqua- drugs were found only trade names.

Either farmers or sellers did not have clear idea about the active ingredients and method

of applications of the particular aqua-drugs though they were using those medicines

without hesitation. It was also seen that same products of different companies had

variable dosages. So, the farmer became confused which product would be effective in

controlling disease. If the causative agents of infectious diseases were identified via a

sensitivity or diagnostic test, it will be very easy to successful administration of

antibiotics with actual dose. However, it is important to follow the recommended dosages

and methods of application of particular antibiotics to get the best results for controlling

disease in aquaculture.

Disease treatment in aquaculture can be a great value when used properly but when

improperly applied, can causes great loses of aquatic ecosystem. So, it is important to

apply drug against appropriate disease and best application methods for aqua-health

management. Pharmaceuticals companies should conduct more research and

development towards reducing the harmful impact of aqua-medicine in aquaculture.

The use of commercial aqua drugs and chemicals in aquaculture for various purposes is

widely now recognized. The aquaculture activities in Bangladesh are also influenced by a

number of chemicals. It is important that policy makers, researchers, and scientists work

together to address the issues of use of new commercial aqua-drugs and chemicals with

the view to reduce their adverse impacts. Efforts should be made towards finding non-

chemotherapeutic solutions to health management and disease control. Also, there is a

need for better understanding of fish farm health management and disease prevention

practices that could reduce the need for drug treatments.

CHAPTER-6

SUMMARY AND CONCLUSION

The research has been done to know the updated status of commercial aqua drugs and

chemicals and their impact on fish health. Data was collected through questionnaire

interview with fish farmers and drug sellers, personal contact and market survey from

selected area in Mymensing district.

Through the observation we obtained different categories of commercial drugs and

chemicals were using for different purposes. From the starting of pond preparation to

disease treatment different drugs and chemicals were used by the farmers. There are

various pharmaceuticals companies were found to provide different types of drugs and

chemicals with different trade name to meet the farmers demand. Different types of fish

diseases like EUS, tail rot, fin rot, red spot, white spot and dropsy in different fish species

mainly in Shing, Koi, Tilapia and Pangus were observed in the study area. To overcome

such kind of situation farmers used different kinds of drugs and chemicals. After use of

drug they got good to better result. But some certain cases they did not get any recovery.

During the field observation some problems were identified in using aqua-drugs which

include poor understanding of farmers about the application of drugs, inadequate

withdrawal period and some adverse effect on fish and human health. It was also found

that all the drugs of a company may not suitable for treatment of disease in a culture

system.

Farmers should have appropriate knowledge about the use of drugs and chemicals. They

also should maintain proper withdrawal period for use of drugs chemicals. The present

research has been only conducted in Mymensingh region and thus other parts of the

country need to be investigated to have a clear picture of the use and impact of

commercial aqua-drugs and chemicals in aquaculture of Bangladesh.

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APPENDICES

Questionnaire used during data collection

Questionnaire for Chemical Seller

Name of Shop:

Name of owner:

Location:

1. What type of chemical you usually sell?

2. Types of aquaculture chemicals available at your store with approximate price.

3. Who is your customer?

4. Do the farmers seek suggestion from you? Y/N

5. If yes, what kind of suggestion you usually offer?

6. Do the farmer satisfied with your products ?Y/N

7. From where you buy you products?

8. Give the following information about chemicals available in your shop. Trade Name Active ingredient Dose Source Price (Tk.)

Questionnaire for Fish Farmers 1. Species of fish cultured:

2. Duration of culture:

3. Do you prepare your pond before releasing fry? Y/N

4. If yes, what is procedure?

5. Did you treat your fish before releasing in your ponds? Y/S

6. If yes, what chemicals and doses do you use?

7. Do you have any disease problem in your farm?

If yes, give details about the disease of your fish.

Disease Clinical sign Species Effective treatment

Prevalence

(%)

Death (%)

Season

8. Do you use chemicals / antibiotics against this disease? Y/N

If yes, Give information of below table:

Trade name

Active ingredient

Purpose of use

Method of application

Dose Price Source

9. How did you know about the particular chemical?

10. Any training opportunities from GO/NGOs on use of chemicals:

11. Do you use common chemicals / antibiotics against all disease? Y/ N

If yes, why?

12. By using chemicals / antibiotics is the fish health condition improved? Y/N

13. From where you take suggestion to use chemicals / antibiotic?