REPLACEMENT OF SOYBEAN MEAL WITH SILKWORM MEAL...

REPLACEMENT OF SOYBEAN MEAL WITH SILKWORM MEAL

(Bombyx mori) IN POULTRY RATION

BY

RAFIULLAH

A dissertation submitted to The University of Agriculture, Peshawar in partial

fulfillment of the requirements for the degree of

DOCTOR OF PHILOSOPHY IN POULTRY SCIENCE

DEPARTMENT OF POULTRY SCIENCE

FACULTY OF ANIMAL HUSBANDRY AND VETERINARY SCIENCES

THE UNIVERSITY OF AGRICULTURE, PESHAWAR

KHYBER PAKHTUNKHWA-PAKISTAN

DECEMBER, 2016



BY

RAFIULLAH

A dissertation submitted to The University of Agriculture, Peshawar in partial

fulfillment of the requirements for the degree of

DOCTOR OF PHILOSOPHY IN POULTRY SCIENCE

Approved By:

_______________________ Chairman Supervisory Committee

Prof. Dr. Sarzamin Khan

_______________________ Member (Major Field)

Dr. Naila Chand Associate Professor

_______________________ Member (Minor Field)

Dr. Nazir Ahmad Khan

Assistant Professor

Dept. of Animal Nutrition

_______________________ Chairperson & Convener Board of Studies

Dr. Naila Chand Associate Professor

_______________________ Dean, Faculty of Animal Husbandry &

Prof. Dr. Nazir Ahmad Veterinary Sciences

_______________________ Director Advanced Studies & Research Prof. Dr. Muhammad Jamal Khan

DEPARTMENT OF POULTRY SCIENCE

FACULTY OF ANIMAL HUSBANDRY AND VETERINARY SCIENCES

THE UNIVERSITY OF AGRICULTURE, PESHAWAR

KHYBER PAKHTUNKHWA-PAKISTAN

DECEMBER, 2016

DEDICATION This humble effort is dedicated to my beloved

Parents and wife whose prayers and encouragement enabled me to achieve this ambition

Rafiullah

TABLE OF CONTETNS

CHAPTER NO. TITLE PAGE NO.

LIST OF TABLES ........................................................................................... i

LIST OF FIGURES .......................................................................................... iii

LIST OF ABBREVIATIONS .......................................................................... iv

ACKNOWLEDGMENTS ................................................................................ v

ABSTRACT ..................................................................................................... vi

I. INTRODUCTION .......................................................................................... 1

II. REVIEW OF LITERATURE ....................................................................... 4

2.1 Silkworm meal in broiler ration ........................................................... 4

2.2 Silkworm meal in layer ration .............................................................. 6

2.3 Insect meal in quail ration .................................................................... 7

2.4 Silkworm meal in the diet of fish and other livestock .......................... 7

2.5 Other insects in poultry ration .............................................................. 8

III. STUDY-1 ......................................................................................................... 15

ABSTRACT .................................................................................................... 16

3.1 Introduction ......................................................................................... 17

3.2 Material and methods ........................................................................... 18

3.2.1 Preparation of Silkworm Meal ................................................. 18

3.2.2 Experimental design, diets and management of

experimental birds .................................................................... 18

3.2.3 Determination of Apparent Metabolizable Energy (AME) ...... 20

3.2.4 Nutrients digestibility ............................................................... 20

3.2.5 Chemical Analysis ................................................................... 21

3.2.6 Organoleptic study ................................................................... 21

3.2.7 Statistical analysis .................................................................... 21

3.3 Results .................................................................................................. 22

3.3. Chemical composition of silkworm meal ................................. 22

3.3.2 Amino acid profile (g/100g of total amino acids) and

mineral profile of silkworm meal ............................................. 22

3.3.3 Effect of replacing soybean meal with silkworm meal on

feed intake, weight gain, feed conversion ratio (FCR) and

dressing percentage of broilers ................................................. 24


apparent metabolizable energy (AME) apparent nutrient

digestibility and organoleptic properties of broiler chicks ....... 24

3.4 Discussions ........................................................................................... 26

3.5 Conclusions .......................................................................................... 28

IV. STUDY-II ........................................................................................................ 29

ABSTRACT ................................................................................................... 30

4.1 Introduction ......................................................................................... 31

4.2 Material and methods ........................................................................... 32

4.2.1 Preparation of Silkworm Meal ................................................. 32

4.2.2 Experimental design diets and management of experimental

birds .......................................................................................... 32

4.2.3 Carcass traits and hematology .................................................. 35

4.2.4 Economics of the different ration (Cost analysis) .................... 35


4.3 Results .................................................................................................. 36


feed intake, body weight gain , feed conversion ratio (FCR)

and dressing percentage of broilers .......................................... 36

4.3.2 Effect of varying replacement levels of silkworm meal for

soybean meal on hematological parameters of finishing

broiler chickens ........................................................................ 37

4.3.3 Economics of replacing soybean meal with silkworm meal

in broiler finisher ration ............................................................ 38

4.4 Discussions ........................................................................................... 40

4.5 Conclusions and recommendations ...................................................... 42

V. STUDY-III ...................................................................................................... 43

ABSTRACT ................................................................................................... 44

5.1 Introduction .......................................................................................... 45

5.2 Materials and methods ......................................................................... 47

5.2.1 Experimental diets and Birds ................................................... 47

5.2.2 Performance parameters ........................................................... 47

5.2.3 Egg quality Analyses ................................................................ 47

5.2.4 Determination of Apparent Metabolizable Energy (AME) ...... 48

5.2.5 Nutrients digestibility ............................................................... 48

5.2.6. Blood profile analysis ............................................................... 49

5.3 Statistical analysis ............................................................................... 49

5.4 Results .................................................................................................. 50

5.5 Discussion ........................................................................................... 55

5.6 Conclusions and recommendations .............................................. 58

VI. STUDY-IV ....................................................................................................... 59

ABSTRACT ................................................................................................... 60

6.1 Introduction ......................................................................................... 61

6.2 Materials and methods ......................................................................... 63

6.2.1 Birds and experimental plan ..................................................... 63

6.2.2 Sample collection ..................................................................... 63

6.2.3 Biochemical analysis ................................................................ 65

6.2.3.1 Liver and renal function tests ....................................... 65

6.2.3.2Histopathological examinations .................................... 65


6.3 Results ................................................................................................. 66

6.3.1 Effect of various levels of soybean meal replacement with

silkworm meal on performance parameters of laying hens ...... 66

6.3.2 Impact of soybean meal replacement with silkworm meal

on liver and renal functions ...................................................... 67

6.3.3 Impact of soybean meal replacement with silkworm meal

on intestinal morphometry ....................................................... 68

6.3.4 Histomorphology of liver, kidney and intestines ..................... 69

6.3.4.1 Histograph of liver tissues ............................................ 69

6.3.4.2 Histograph of kidney tissues ........................................ 70

6.3.4.3 Histograph of the intestines .......................................... 71

6.3.4.4 Histomorphological criteria ......................................... 72

6.4 Discussion ............................................................................................ 73

6.5 Conclusion ............................................................................................ 75

VII. SUMMARY, CONCLUSIONS AND RECOMMENDATIONS ................ 76

7.1 Summary .............................................................................................. 76

7.2 Conclusions .......................................................................................... 81

7.3 Recommendations ........................................................................ 81

LITERATURE CITED .................................................................................. 82

i

LIST OF TABLES

TABLE NO. TITLE PAGE NO.

3.1 Ingredient and chemical composition of experimental rations for broiler at

starter phase ...................................................................................................... 19

3.2 Chemical composition of silkworm meal ........................................................ 22

3.3 Amino acid profile of silkworm meal .............................................................. 23

3.4 Minerals profile of silkworm meal .................................................................. 23

3.5 Effect of replacing soybean meal with silkworm meal on feed intake,

weight gain, feed conversion ratio (FCR) and dressing percentage of

broilers .............................................................................................................. 24

3.6. Effect of replacing soybean meal with silkworm meal on apparent

metabolizable energy (AME) of broiler chicks ................................................ 25

3.7 Apparent nutrient digestibility of broiler starter chicks fed with different

levels of silkworm meal ................................................................................... 25

3.8 Organoleptic quality of broiler meat provided silkworm meal diets ............... 26

4.1 Ingredient and chemical composition of experimental rations for broiler at

starter phase ...................................................................................................... 34

4.2 Effect of replacing soybean meal with silkworm meal on feed intake, body

weight gain, feed conversion ratio (FCR) and dressing percentage of

broilers .............................................................................................................. 36

4.3 Effect of varying replacement levels of silkworm meal for soybean meal

on Hematological and carcass characteristics parameters of finishing

broiler chickens ................................................................................................ 37


carcass characteristics parameters of finishing broiler chickens ...................... 38

4.5 Economics of replacing soybean meal with silkworm meal in broiler

finisher ration ................................................................................................... 39

5.1 Ingredient and chemical composition of experimental rations for layer .......... 51


on performance parameters of laying hens ...................................................... 52

ii


on digestibility coefficient of nutrients of laying hens ..................................... 52


on Hematological parameters of laying hens ................................................... 53


on egg quality parameters of laying hens ......................................................... 54

6.1 Ingredient and chemical composition of experimental rations for layer .......... 64

6.2 Effect of various levels of soybean meal replacement with silkworm meal

on performance parameters of laying hens ...................................................... 66

6.3 Impact of silkworm replacement in feed on liver and renal functions ............. 67

6.4 Impact of soybean meal replacement with silkworm meal on intestinal

morphometry .................................................................................................... 68

6.5 Histomorphological criteria ............................................................................. 72

iii

LIST OF FIGURES

FIGURE NO. TITLE PAGE NO.

1. Histomicrograph of liver tissues (1, 2, 3, 4, 5) ................................................. 69

2. Histomicrograph of kidney tissues (6, 7, 8, 9, 10) ........................................... 70

3. Histomicrograph of the intestines (11, 12, 13, 14, 15) ..................................... 71

iv

ACKNOWLEDGMENTS

I am extremely thankful to Almighty ALLAH, The beneficent and merciful who helped me

a lot in every moment of my life. I offer my humblest thanks to The Holy Prophet “Hazrat

Muhammad”ملسو هيلع هللا ىلص who is forever a source of guidance and inspiration for humanity.

Firstly, I would like to express my sincere gratitude to my advisor Prof. Dr. Sarzamin Khan

for the continuous support of my Ph.D study and related research, for his patience,

motivation, and immense knowledge. His guidance helped me in all the time of research

and writing of this thesis. I could not have imagined having a better advisor and mentor for

my Ph.D study.

I would like to express my deep gratitude to my worthy and honorable Prof. Dr. Nazir

Ahmad, Dean Faculty of Animal Husbandry and Veterinary Sciences, The University of

Agriculture, Peshawar for his indispensable guidance and valuable support had provided

me a good basis for my report and research work.

Besides my advisor, I would like to thank the rest of my thesis committee, Prof. Dr.

Muhammad Subhan Qureshi, Dr. Naila Chand, Dr. Asad Sultan, Dr. Syed Muhammad

Suhail and Dr. Zahoor Ul Hassan for their insightful comments and encouragement, but

also for the hard question which incented me to widen my research from various

perspectives.

Also I thank my teacher in Department of Animal Nutrition in particular, I am grateful to

Dr. Nazir Ahmad Khan (Assistant Professor) for enlightening me the first glance of

research.

My sincere thanks also goes to Prof. Dr. Umar Sadique, who provided me an opportunity to

join their team, Dr. Said Sajjad Ali Shah and Dr. Hayatullah and other colleagues and gave

access to the laboratory and research facilities. Without their precious support it would not

be possible to conduct this research.

I thank my fellows Dr. Naseer Ahmad, Dr. Kamran Khan, Dr. Muhammad Inam, Dr. Nadar

Khan, Dr. Shahid, Dr. Waqas, Dr. Bilal and Mr. Raees Khan for the stimulating

discussions, for the sleepless nights when we were working together before deadlines, and

for all the fun we have had in the last four years.

Last but not the least, I would like to thank my family: my loving parents, brothers, sisters

and especially my beautiful wife for supporting me spiritually throughout writing of this

thesis and my life in general.

Rafiullah

v



Rafiullah and Sarzamin Khan

Department of Poultry Science, Faculty of Animal Husbandry and Veterinary Sciences

The University of Agriculture, Peshawar-Pakistan

December, 2016

ABSTRACT

A series of research trials were conducted to study the effect of replacing soybean meal

with silkworm meal on production performance, apparent metabolizable energy

(AME), apparent nutrients digestibility, haematology and carcass traits in broiler during

starter and finisher phase. Five iso-nitrogenous and iso-caloric rations were set on the

basis of chemical composition of silkworm meal as 0% (D1), 25% (D2), 50% (D3),

75% (D4) and 100% (D5) substitution of soybean with silkworm meal in broiler ration.

The silkworm meal analysis showed higher levels of CP (57.6 ± 1.50%) and crude fat

(23.3 ± 0.21%). The silkworm meal had higher contents (g/100g of total amino acids)

of lysine (7.52 ± 0.376) and methionine (3.88 ± 0.376), but was slightly deficient in

tryptophan (1.70 ± 0.413). In first trail total 250 day-old broiler chicks were divided

into twenty five replicate groups (n=10), with five replicates per diet using a completely

randomized design. Performance parameters such as feed intake, live weight and

dressed(%) were higher (P < 0.05) in group D4. Moreover, lower (P < 0.05) FCR was

observed for D1 and D4 groups than D3 group. AME was higher (P < 0.05) in group

D4 and D5 in comparison with other groups. The digestibility of DM, CP, crude fat,

ash and N free extract did not differ (P > 0.05). The organoleptic quality parameters

were not altered (P > 0.05) due to replacement of soybean meal by silkworm meal. The

findings of the present study revealed that silkworm meal may effectively be used

replacing 75% soybean meal as low-cost protein ingredient in broiler starter ration.

Second trail was conducted on replacing soybean with silkworm meal in commercial

broiler finisher ration. Total 150 day-old chicks were randomly divided into fifteen

replicate (n=10), and consequently reared on five experimental diets according to a

completely randomized design. Feed intake, weight gain and dressing percentage varied

(P < 0.001) due to diet composition, and the highest (P < 0.05) values were recorded for

diet D4. Moreover, numerically lowest FCR was observed for D1 (control) and D4

diets, whereas the FCR for D2, D3 and D5 diets was not different (P > 0.05). The

dressing percentage was not significantly different (P > 0.05).However, no significant

(P>0.05) variation in blood parameters and carcass traits was observed among dietary

treatments.Price tag per kg of feed gradually decline and lowest price per kg of meat

and highest profit per kg meat was recorded for diet D4. It was revealed that silkworm

meal may be effectively utilize as low price animal protein constituent in broiler

finisher ration.

Third experiment was conducted to study the effect of soybean meal replacing with

silkworm meal in layer. Five iso-nitrogenous and energetic diets were developed as 0%

(D1), 25% (D2), 50% (D3), 75% (D4) and 100% (D5) replacement of soybean meal

with silkworm meal.Separate feeders and drinkers were installed in each cage for the

provision of water, feed and assigned normal condition throughout the trail period. A

total of 150 laying birds were randomly divided into fifteen replicate groups (n=10),

and reared on five experimental diets according to a completely randomized design.

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The weight of the bird, daily feed intake (g/day/bird), average egg weight, hen day

production (%) feed conversion ratio (g feed/dozen egg production), did not differ

significantly (P 0.05) in the control group compared to group D2, D5 and

D3Blood profile and corporal parameters of egg showed no significant differences

among dietary groups. The egg weight, yolk weight, shell thickness and albumen height

were not affected (P>0.05) with dietary treatment. It was assessed that silkworm meal

as alternative protein constituent can be effectively used in layer ration without any

adverse effect.

Fourth trial was conducted to probe any adverse outcome of the soybean meal

replacement with silkworm meal on serum and histomorphology of various organs in

laying hens. A total of 75 white leghorn birds were allocated into five dietary groups

with three replicates in separate cage. Five rations were formulated according to

nutrients requirements of the layers in which soybean meal were step-wise (0, 25, 50,

75 and 100 %) replaced with silkworm meal. For the analysis of biochemical

parameters and histomorphological measurements of organs six (6) birds were

randomly selected per treatment at the end of trial. The results showed that bird weight,

daily feed intake (g/day/bird), hen day production (%), average egg weight (g) and feed

conversion ratio (g feed/dozen egg production) did not differ significantly (P 0.05) during the experimental period. The

height and thickness of villous, and goblet cells number were increased numerically in

treated groups as compared to control. It was concluded that replacement of varying

level of soybean with silkworm meal in laying birds diet had no hostile effect on

production performance, intestinal histomorphology, biochemical functions of liver and

integrity of vital organs. In short silkworm meal is a potential alternative of soybean

meal for both broiler and layer ration improving production performance with no

toxic effect on health of birds which may be committed as a solution for

entrepreneurship development under Dairy Science Park.

Key words: Soybean, silkworm, broiler, layer, performance, egg quality,

biochemical analysis, histomorphology

1

I. INTRODUCTION

The worldwide food production systems are facing increasing competition due

to human population increasing demand of food source sowing to increasing income

level and concern about human health (Makkar et al., 2014). The meat demand will be

up raise from the developing countries of the world as compared to 2010 and in the

future 2050 the demand will be 58 percent (FA0, 2011).The feed takes the largest part

of the animals production costs, and most of it is contributed by the cost incurred on

protein ingredients, particularly in monogastric animals such as poultry. The latter

could not synthesize essential amino acids and need dietary sources of these amino

acids to be able to achieve optimum production and reproduction efficiency.

Poultry meat contributes approximately 37% of the total animal protein supply

in most developing countries and there is the possibility of growth and expansion of

this industry (Ahmed and Islam, 1990. Broiler (meat type chickens) producers are

facing much difficulty with availability and higher prices of feed ingredients (Khatun et

al., 2003). Feed materials, especially animal origin protein are high in price value and

rare in availability due to competition with human, poultry and other animals etc.

Poultry industry primarily dependent on plant origin based feed ingredients particularly

soybean meal due to its excellent amino acid profile and higher digestibility value

(Willis, 2003).

In many countries such as Pakistan the production of soybean is limited and as

such most of the soybean used in poultry ration is imported from other countries. The

cost of soybean has significantly amplified in a few years due to the rapid growth of

poultry industry on global basis, as well as by the rising demand of soybean in the

human food industry. The increasing cost of protein ingredients has stimulated the

reason of low cost protein ingredients for poultry ration(Ramos- Elorduy et al., 2002;

Das et al., 2009). Recently there is a growing interest in the use of some protein-rich

insect species as a low-cost protein ingredient in poultry ration (Van-Hhuis et al.,

2013). Insects nurture and breed very fast, had high feedstuff alteration effectiveness

and can be fed on bio-wastes materials. State-of-the-art studies have revealed that it is

quite practicable to harvest insects on a huge scale and be used as substitute and

maintainable protein constituent in poultry feed (Veldkamp et al., 2012), and the

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benefits could be particularly greater, whenever if they provide substrate of bio-waste

or organic tributaries.

Insects have great nourishing value in proteins, fats, vitamins and minerals

(Chapman, 1998). The value of protein of edible insects ranges from 30% to 80%

(certain wasp species). Insects have a diverse mineral similar or higher than the

foreseeable feeds.

Protein-rich insects are the best option to decrease the price tag of protein

additions in poultry feed and improve food and food safety (Van Huis et al., 2013).

Insects have been used as a diet basis for different species of animal. However less

study have been conducted on the use of insects in poultry ration. Mostly insect’s

species are used as feed in a wide range of animals. It can easily reproduce and grow

with good efficiency because these insect can be reared on the wastes products easily

due its habitat nature. Silkworm caterpillar hatches from a tiny black egg laid by the

adult moth. It feeds on mulberry and shear butter leaves constantly and grows to full

size of 7.5-10cm within 4-6 weeks (Enchanted learning, 2005). In larval phase of

silkworm forms a protective sheath of silk, and at the last of this phase pupa of

silkworm with the help of enzyme the protective sheath break and moth is out of it

which is the main waste of silk production (Datta, 2007).Once selected, using

appropriate breeding means, appropriate species offer a consistent and viable source of

good quality protein for feeding of poultry (Anand et al., 2008).

Silkworm, which can be produced as co-product of the silk industry, is a

caterpillar of moth butterfly (Bombyx Mori) whose cocoon is used for silk production.

Owing to its good quality natural protein and better digestibility silkworm pupae has a

good and alternative livestock feed, markedly for monogastric animals (Trivedy et al.,

2008). Silkworm caterpillar rich in protein (56.8-63.8%), lipid (19.6-31.5%) and ash

(3.3-7.7%) (Loselevich et al., 2004; Makkar et al., 2014). Apart from these nutrients, it

has an amino acid profile which in most cases compares favourably with those of fish

meal (Solomon and Yusufu, 2005). However, according to our knowledge, limited

work has been done on the nutritional point of silkworm caterpillar meal produced in

Pakistan as animal feed.

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Consumed silkworm pupae are surplus material and are cast off to the open

environment or on the other hand castoff as bio manure (Wei et al., 2009).According to

the statement of (Trivedy et al., 2008) oil to be haul out as valued industrial product in

plastics, soaps, varnishes, paints, biofuels and candle etc.

Silkworm meal as human diet in silk producing countries mostly in Asia and is

pondered as a fragility in areas of China (Luo, 1997), Thailand (Yhoung- Aree et al.,

1997) and in India (Longvah et al., 2011). Silkworm pupae plunders quickly owing to

greater water content. Silkworm meal can be stored for longer time whenever oil are

extracted and defatted. Moreover, the defatted meal has complex protein gratified as

(Blair, 2008).

Caterpillar meal is high biological value rich in protein (56.8-63.8%), lipid

(19.6-31.5%), crude fiber (0-5.8%) and ash (3.3-7.7%) (SN1, 2005; Loselevich et al.,

2004). Apart from these nutrients, it has an amino acid profile which in most cases

compares favorably with those of fish meal (Solomon and Yusufu, 2005). Exploration

effort on silkworm caterpillar meal as food component in Pakistan is scarce.

Therefore this study is premeditated to estimate the impact of silkworm

caterpillar meal on the performance parameters of broilers and layers in comparison

with soybean meal with the following objectives:

1. To assess the mineral and amino acids profile of silkworm meal in comparison

with soybean meal.

2. To examine the varying replacement levels of silkworm meal on soybean meal

and its impact on growth performance, carcass traits, hematology and nutrient

digestibility by broiler birds at different phases of life.

3. To evaluate the varying replacement levels of silkworm meal on soybean meal

on laying performance, egg quality traits, nutrients digestibility and hematology

of laying birds.

4

II. REVIEW OF LITERATURE

Literature on the silkworm meal and related alternate protein sources and their

uses in poultry ration is reviewed in this chapter under the following subtitle.

2.1 Silkworm meal in broiler ration

Agbede and Aletor (2003) performed a research trail in which fish meal was

replaced by glyricidia leaf a substitute of protein in feed of chicks. Earlier to trial, the

glyricidia leaf was investigated for the amino acids and other chemical contents. On the

basis of chemical composition five diets namely control (5% fish meal) and in rest of

four diets glyricidia leaf was added at 25, 50, 75 and 100% in diet. A total of 300

chicks were allotted into five dietary treatments in triplicate. After the completion of

the trial both the body weight gain and final body weight were found similar to that of

the control group feed. The glyricidia leaf level in the diet was found inversely

proportion to the feed consumption and weight gain. Chicks utilizing 1.81, 3.62 and

5.43% glyricidia leaf based feed were found similar in withholding ability of nitrogen

with that of control group. The Erythrocyte Sedimentation Rate(ESR) were observed

directly proportional with the elevated level of glyceridea leaf based diet whereas the

no alteration occur in the serum component due to dietary treatment. This study showed

that the lowest level of glyricidia leaf as a protein supplement can be replaced by fish

meal without any harmful effect on bird performance.

Khatun et al. (2003) carried out a research trial in broiler chicks in which

silkworm pupa meal (SWPM) was incorporated into the diet for fish meal. Four (iso-

calroic and iso-nitrogenous) experimental diets such as 6 (%) FM+ 0% SW meal, (4%)

FM + 2(%) SW meal, 2 (%) FM + 4(%) SW meal and 0 (%) FM + 6 (%) SW meal

were prepared. It was found that, feed conversion ratio, growth performance, carcass

produce and profitability improved with the increasing level of silkworm meal.

Ijaiya and Eko (2009) studied the effect of replacing silkworm caterpillar meal

(SCM) for fish for potential growth, nutrient utilization via economics of day old anak

broiler chicks. A total of one hundred and fifty (n = 150) chicks were allotted randomly

to five dietary treatment groups. Each treatment group contain 30 birds having two

replicates containing 15 birds per replicate. The control diet contain 100% fish meal

5

whereas diet 2, 3 and 4 SCM was replaced by 25, 50 and 75%, respectively and diet 5

contain no SCM and fish meal. The composition of the feeds were nutritionally

maintained both in energy and protein content. All the birds were provided the feed and

drinking water ad-libitum. Feed intake, body weight gain, feed conversion ratio, protein

efficiency ratio and nutrient utilization of the birds did not alter significantly among the

treatment groups. With increasing dietary level of SCM the cost per kg gain decline

gradually that showing high economic return. The study verified that SCM can be an

outstanding and cheaper replacer for fish meal while formulating ration for starter

broiler chicks in terms of economic gain.

Ijaiya and Eko, (2009) studied the outcome of substituting of fish with silkworm

meal on the growth potential, hematology carcass traits and economics production of

one hundred fifty (n = 150) anak broiler (4 weeks old). The five treatment groups (such

as 100% fish meal: 0% SCM (serving as control), 75% fish meal, 25% SCM, 50% fish

meal, 50% SCM,25% fish meal, 75% SCM and 0% fish meal, 100% SCM) were put in

randomized complete block design with each group containing thirty birds. Each

treatment was blocked into two sub groups containing 15 birds per replicates.

Experimental birds were provided feed and water ad-libitum. There were no significant

difference (P > 0.05) observed amongst the different experimental groups in feed

consumption (95.7 - 98.2 g), body weight gain (46.1 - 98.5 g), feed ratio ranges (1.98-

2.08) and protein competence ratio ranges 2.41 - 2.54. Similarly, the value of carcass

weight and body cuts was non-significant. Analysis of the blood parameter showed

non-significant (P > 0.05) difference among the treatment groups except albumin. The

economic efficiency indicated that cost/kg of feed gradually reduced as the SCM

dietary levels increase resulting in high economic return. Overall the study

demonstrated that the growth potential of the broiler birds was not alter due to the

inclusion of SCM. However, it was more cost effective than that of fish meal and can

be alternative substitute for fish meal in broiler during the finishing phase.

Dutta et al. (2012) determined the result of various levels of SWPM (Silk worm

pupa meal) with expensive fish meal in Rohd Island Red (RIR) strain (age, 3 days old)

of birds. The chicks were divided into five dietary treatment groups of SWPM.

Proximate analysis was used to find out the energy cost and from the growth potential

of chicks receiving different level of SWPM. This study suggested that SWPM is the

6

most economical source and has the potential protein source to be replace on the fish

meal in the diet of poultry birds.

Oso et al. (2014) performed experiment owing growing performance and

ingredient utilizations of Gariepinus juveniles on Bombyx Mori meal (BMM) as

substitute for fish meal. The varying level of insertion in dietas B (25%), C (50%), D

(75%) and E (100%) correspondingly of five calculated experimental diet. The diet A

(Control) was 100% fishmeal and nurtured two times daily at 5(%) of the body weight.

The fish were collected from private hatchery at Ado Ekiti for experiments and stocked

in plastic container having capacity of 20 liters with stocking density of 10 fingerlings

per container. The diet effect on feed consumption, growth performance, mineral and

carcass composition of fish were noted. The outcomes of the diets showed specific

growth rate, weight gain and efficiency of protein ratio of fish feed in the diets were

significantly (P

7

improvement in layer efficiency was observed in SWPM consuming birds with that of

control group. In a nut shell it was concluded that SWPM might be an excellent source

of protein and a good replacer of costly protein source in layer ration ultimately

enhance the profitability of layer birds.

2.3 Insect meal in quail ration

Widjastuti et al. (2014) evaluated that black solider fly (BSF) maggot as an

excellent source of crude protein (46.6%), and a good alternative of imported fish meal

in the poultry diet. The BSF maggot is the larvae of Hermitia illucens (hatch in four

days), and contain 4.32% crude fiber, crude fat (23.6%) and 3457 kcal/kg

metabolizable energy. Its amino acid content is equivalent to that of fish meal. They

replaced the fish meal with the BSF maggot meal in the ration of Japanese quail. A

total of one hundred (n = 100) female Japanese quails one and half month were reared

in cages for up to 5 months using RCBD design. The birds were grouped based on their

diet and within each group four replicate were made in such a way that birds in group 1

fed 100% fish meal, while in rest of the diet fish meal was replace by BSF @ 25 (%),

50 (%) and 100 (%), respectively. The outcomes of feed intake, feed conversion ratio

and egg weight differ significantly with the substitution of fish meal protein with BSF

protein in the quail diet. However, no significant difference among treatment groups

was noted in egg production. The average feed consumption was higher in group 1 as

compared to quail of 100% BSF level. It was concluded that the BSF maggot meal is a

good protein source in quail ration and can be used as an alternate to the fish meal

protein.

2.4 Silkworm meal in the diet of Fish and other livestock

Ogunji et al. (2008) studied the consequences of changing the fish meal in the

feed of livestock and fish having protein of animal and plant origin. Unfortunately

efforts to practice these components for whole replacement of the fish meal constituent

in tilapia diets have not completely been fruitful. This might be due to the lower level

of essential amino acid content in the diet particularly methionine. Megmeal appear to

be a potential candidate for the substitution of fish meal due to the cost effectiveness,

more availability, the content of crude protein and its amino acid profile. For this

purpose the study was conducted to formulate seven different diets by replacing fish

8

meal with magmeal and it was then offered to the fifteen fingerlings. The fingerlings

were divided in triplicate for 56 days. Results showed no significant difference among

all dietary treatment groups for growth traits, protein utilization, stress indicators and

hematological parameters. These findings recommended that fishmeal can be replace

completely by magmeal, as magmeal can meet all the nutrient requirements of Tilapia

(Oreochromis niloticus) fingerling, and can be included in the ration.

Olaniyi and Babasanmi (2013) conducted research on the substitute of fish meal

on silk worm pupae in the diet and used as protein source for Clarias gariepinus, of

African cat fish. The fingerlings (initial body weight 14.3 g) were divided into five

dietary treatment (40% content of crude protein) groups. The silk worm meal was

included at the level of 100, 75, 50, 25 and 0% (control, 100% fish meal) to the fish

diet. In 12 weeks study, protein utilization, feed conversion, specific growth rate, final

body weight and proximate composition were noted. The highest proportion of silk

worm pupa meal significantly (P < 0.05) enhanced the growth potential and efficiency

of feed conversion of fingerlings. It was concluded from their research that there is a

significant differences (P < 0.05) found in all the studied parameters along with dietary

treatment. However, the inclusion levels of 50 and 75% did not exert any significant (P

> 0.05) variation for growth traits. It was recorded that 100% proportion of silk worm

pupa meal indicated variations in feed proficiency, deploy and utilizing protein for the

potential growth and body weight gain. No significant variation for the proximate

composition of fish fed 75, 50 and 25% inclusion level, while the contents of crude

protein and lipids increased at the initial phase of the trial. These findings suggest that

the highest (100%) inclusion level of silk worm pupa meal can be used as a source of

protein and a good alternate of fish meal in the diet of fingerlings.

2.5 Other insects in poultry ration

Yaqub (1991) performed experiment for the productions of maggots using the

animal carcass and poultry droppings as a substrate. The harvesting times for maggots

were 5 to 8 days. After harvesting the maggots were removed from the substrate by

means of mesh sieve (5 mm size) and collected in trays. Maggots along with the

substrates were placed on the sieve and put over the on the sieve and dried in the sun.

The sieve is placed in such a way that maggots pass it easily and collected in the tray.

The flies’ maggots cross away the filter and collected by the hanging tray in the

9

container. After collection, the maggots were placed in the sun for drying. Finally, sun

dried maggots were placed in the oven at 50 oC for 24 hours for complete drying.

Akpodiet et al. (1998) utilized the feces of layer birds for the maggots’

production. Approximately 1 kg dropping per fowl were collected in a plastic bowl.

After collection, the dropping were mixed with palm oil (100 ml) subsequently. The

bowl was placed in open shed where the houseflies oviposited. On daily basis the

cultures were moistened adequately with sprinkling water for 6 days before larvae

harvesting to support the larvae growth. At the time of harvesting, to each bowl plenty

of water was added to totally to the feaces in which maggots can float. The produce

maggots were harvested, filtered, washed and unfettered of the remains. After complete

harvesting maggots were kept on fire for drying and crushed with hammer mill and

added in the diet. The maggots were than harvested, filtered, washed, and unfettered of

debris. After complete harvesting, the maggots were placed on fire for roasted dried

and crushed with in a laboratory hammer mill and then assimilated into the diet of

poultry.

Awoniyi et al. (2004) studied the comparison effect of fish and maggot meal. A

total of 90 broilers at the age of four weeks were used in the experiments. They are

divided into five groups with 18 birds per replicate in which maggot meal (MGM)

replaced at 0,25,50,75, and 100% with fish meal. The samples were collected from each

replicate and subjected to total erythrocytes counts (TEC), packed cell volume (PCV),

mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration

(MCHC) and mean corpuscular volume (MCV). Result showed no significant

difference (P 0.05) among the treatments. They found no changes in erythrocyte indices

and concluded that bacterial micro flora from maggot meal sample were similar to fish

meal sample and no adverse effect on blood physiology and health status of chicken.

The result of this experiment showed that (MGM) can be replaced with fish meal

without effectively having no direct effect on the blood parameters of birds and

indirectly on health of consumers of chickens.

Adenije et al. (2008) studied the importance of rumen content maggot meal

(RCMM) rumen blend and maggot’s meal. This study was conducted in which the

RCMM were fed to piglet weaned at 28 days of age. The RCMM was included at level

of 0, 5, 10, 15 and 20%. Increasing the level of RCMM in the diet showed no

10

significant effect (P > 0.05) on weight gain and fed to gain ratio of weanling piglet.

However, the treatment had significant (P < 0.05) impact on fed intake. There was also

no treatment effect (P > 0.05) on nitrogen intake and nitrogen digestibility. The result

of this experiment concluded that maggot meal mixed ration had no negative impact on

the growth rate and other characteristic of weaned piglet. Furthermore he found that

maggot meal mixed diet is economical.

Ogunji et al. (2008) utilize magmeal (housefly) in the diet of tilapia fingerling

and total of 7 iso-energetic diets (crude protein content 36%; and 20 KJg-1

gross energy

on dry matter basis) were formulated by substituting fish meal by magmeal. Total 15

fingerlings (average initial body weight, 2.0 ± 0.1 g) were divided in each experimental

tank were fed at 5% of their body weight in triplicate twice per day. Feed conversion

efficiency and growth rate between feeding groups were similar and satisfactory

without showing any significant variation. According to nutrient profile analysis,

magmeal had an excellent source of amino acid content as compared to fish meal.

However, it has a higher (19.8%) fat content which not only influenced the total fat

contents but also the composition of the fatty acid of diet and fish as well. Optimum

growth of Oreochromis niloticus fingerlings resulted with diet comprising magmeal as

a main source of protein, equivalent to that attained with fish meal. However, the

essential fatty acid are recommended to be included in the diet of magmeal to improve

the optimal fatty acid profile required for proper metabolic functions.

Hwangbo et al. (2009) assessed broiler growth and meat quality with utilization

of maggot meal in the diet. A total 600 broiler chicks were randomly grouped in 5

treatment groups. Each group consisting 40 replicates having 3 birds/replicate. The

birds were reared on different diets as 0% (Control), 5%, 10%, 15% and 20% maggots

etc. Supplementation of maggot to the basal diet resulted an increased in body weight.

Though, no changes found in the feed conversion efficiency. The performance of

broiler chicken was mostly dependent on the optimum profile of amino acid, high

(64.0%) protein content, essential amino acid content (29.5%) and higher (98.5%)

protein digestibility of the maggots. The efficient diets on the return of body weight

improvement was that having 10 and 15% maggots for broiler chicken (age, 4-5

weeks). On the other hand, it also improved significantly the dressing out percentage,

thigh and breast muscle. However, no significant variation was noted for liver,

11

abdominal fat, meat colour and the content of crude protein. Compared to control diet,

the maggot supplemented feed the content of lysine and tryptophan levels of breast

muscle increased significantly in the birds. The feeding diets containing 10 to 15%

maggots in chicken can improve growth and meat quality.

Odesanya et al. (2011) discussed the importance of maggot meal in poultry

ration. They studied that poultry industry is a serious problem of shortage of protein

sources and mainly the only source of protein is soybean and fishmeal which is very

costly and that’s why increases the cost of this sector. Therefore they used alternative

source of protein like maggot meal. The proximate composition of the maggot meal

revealed that the content of crude protein was 48.0%, moisture (86.0 ± 0.47%), ash

(10.0 ± 0.44%), crude fiber (5.89 ± 0.05%), crude fat (31.8 ± 0.02%) and Kilo Calories/

Kg was 3755 ± 190. The fat analysis showed uric acid content 69.9%, stearic acid

12.8%, palmitic acid 2.09% and oleic acid 15.3%. Moreover, the colorimetric

determination of the content of amino acids resulted lysine and methionine 5.03 and

2.58%, respectively. This meal has high nutritive profile and can be used poultry ration

effectively.

Moreki et al. (2012) conducted a trail on feed costs evaluation and reported

almost 70 to 80% total costs of production was due to the importation of the feed

components used in Botswana. Due to high cost of the feed most of the poultry farmers

failed to fed their birds due to reduced growth rate and weight gain which lead to

product loses. This background showed that an alternate source such as insects to be

used as a source of protein which was the most feasible and easily available source

throughout the year. In Botswana, Imbrasia belina cast-off good quality protein in the

poultry ration. This study shows that different insects i.e. fly maggots, gross hopper and

cricket may be a substitute of fish meal in poultry diet based upon the protein content.

The addition of maggot meal likely to lower the cost of feeds in poultry diets, thus

contributing to the profitability of smallholder poultry production. Harvesting insects

for use as feed ingredients will reduce damage in the crop fields, minimize the use of

pesticides for management of pests and reduce environmental pollution. However, the

replacing of fish meal with maggot meal as a source of protein in the poultry ration

different factors like the seasonality aspect, as well as, the presence of chitin which

detrimentally affects protein digestibility must be considered. Further investigations on

12

the nutritional composition of insects used for human consumption should be

conducted with a view to utilizing them in livestock nutrition as well.

Okah and Onwujiarir (2012) studied effectiveness of maggot meal as a replacer

for fish meal in the broiler chicks feed at the finisher stage. On the basis of the maggot

meal analysis CP (44 %) CF (10 %), Fat (2%), ash (14%) NFE (17%), Ca (0.03%) and

P (0.05%).There were 5 feed formulations in which 0, 20, 30, 40 and 50% of the fish

meal were replaced with maggot meal in each group respectively. The diet contained

maggot meal (40%) and (50%) showed lower feed intake but high weight gain as

compared to the control group. Overall the birds fed with maggot meal showed greater

weight gain than that of fish meal. Feed conversion ratio was improved in chicken fed

maggot meal diets than those fed the control diet. Addition of maggot meal in the ration

is very cost effective to diet contained fish meal. The diet contained (50%) maggot

meal FCR was found 34.2 cheaper as to the birds fed with normal diet. The maggot

meal addition in diet particularly 30% and 40% has a significant change in the

deposition of fat in the inguinal region. In aunts shell the replacement of 40% dietary

maggot meal and fish meal showed excellent performing and also proved to be a more

economical and cost effective during the finishing phase.

Tabinda et al. (2013) assessed the suitability of chicken intestine in the feed of

fish fingerling (Cirrhinus mirigala) as substitute protein. The proximate composition of

intestine was performed and five diets were made on the basis of substitute protein as

(FM0), (FM25), (FM50), (FM75) and (FM100) as control diet. The diets were iso-

caloric on the basis of protein, lipids and energy. A trial was planned in control

environment and found parallel growth performance of (FM75) and (FM100).

However, FM50 have higher growth to FM75 and Control. It was concluded that FM25

(P

13

transportation. Embryos of the housefly were preserved at low temperatures of 4-10oC

on moist sponge in petri plates and also stored in water at 26oC. The lowest embryo

survival rate (45%) was observed aging 0-3 hrs at 5oC for 24 hrs. Embryo (aged, 3–12

h) had no negative impact at 5 oC for 24 h; however longer storage caused significantly

decreased in hatchability rate that is 30–34% after 48–72 h, larval survival rate as

compared to control (61%) group, and also decreases to 83 percent when stored after

seventy two hour. It was observed that a lower temperature 10oC for 24 hrs have no

negative effect on embryo aged 0-9hrs. High mortality was observed at 26oC for 24 hrs

in all the groups.

Kenis et al. (2014) surveyed numerous part of the world, especially in West

Africa found that more than 70% of the farming cost is due to the protein contents in

the feed in livestock and fish farms. Due to high cost of the feed ingredient especially

protein it is not properly available for small scale fish and poultry farms. Food and

agriculture organization strongly promoted a practice to use insects in the feed of

poultry and fish farming, as a tool for poverty alleviation. Insects, which are a natural

food source can be used to improve animal diets due to its richness in content of protein

and other valuable nutrients. The usage of insect’s meal in fish diet by fish farmers in

animal diet at West Africa for animal source protein. Flies particularly, black soldier fly

(Hermetia illucens), common house fly (Musca domestica) and black soldier fly

(Hermetia illucens) are the most favourable insects, that can easily be mass in small

production units on farm for domestic use or at industrial level at the community. Flies

have the benefit over many other insects for developing on easily and freely obtainable

waste material and could even contribute to rural sanitation. Small holder farmer

traditionally used termites to feed in backyard poultry. However, their mass production

is challenging, approaches to increase population’s on-farm and assist collection can be

developed. There is a growing need of new techniques to show their economic

profitability, social acceptability and eco-friendly sustainability.

Rahnamaeian et al. (2015)studied protein components of insects as

antimicrobial peptides as immune system in set against pathogens. Against Escherichia

coli Abaecin displaced no detectable activity when tested alone at concentrations of up

to 200 mM, But only at concentrations greater than 2 mM whereas hymenoptaecin

affected bacterial cell growth and viability. The higher the bactericidal effects on

hymenoptaecin in grouping as 1.25 Mm abaecin, for understanding their interaction,

fluorescence resonance energy transfer and atomic force microscopy based assays were

14

used for determination of mechanisms of action. To minimize the level of inhibition,

concentration of hymenoptaecinabaecin was found whenever membrane composed of

hymenoptaecin which interact with chaperone DNA of bacteria. Gram negative

bacterial pathogenic organism develop resistance to antibiotics, therefore various

combinations of antimicrobial pathogens may be used for various treatments

experimentally. These naturally occurring potentiating interactions suggested.

Khan et al. (2016) studied the importance of insect meal in poultry ration. For

this purpose, the expensive soybean meal was replaced through maggot meal. In this

trail 120 chicks were separated into four different treatment. The diets were calculated,

such a way that soybean meal was replaced by maggot meal at different level as 0 %

(A), 10% (B), 20% (C) and 30% (D) in diet respectively. The birds were allowed to

water and feed ad-libitum throughout the experimental period. There was a high feed

efficiency, body weight gain and dressing percentage containing 30% maggot meal

while the Feed consumption reduced significantly in all treated groups. Likewise,

obvious metabolizable energy also enhanced significantly in the 30% fed group;

however, on the other hand, dry matter, crude protein, crude fiber, ether extract, ash and

nitrogen free extract did not differ significantly between treated and control groups.

Findings of the sensory analysis showed that juiciness and tenderness along with

palatability significantly improved in B and C group respectively.

15

CHAPTER-3

STUDY-1

EFFECT OF REPLACEMENT OF SOYBEAN MEAL BY

SILKWORM MEAL ON GROWTH PERFORMANCE, APPARENT

METABOLIZABLE ENERGY AND NUTRIENTS DIGESTIBILITY

IN BROILERS AT DAY 28 POST HATCH

16

EFFECT OF REPLACEMENT OF SOYBEAN MEAL BY SILKWORM MEAL

ON GROWTH PERFORMANCE, APPARENT METABOLIZABLE ENERGY

AND NUTRIENTS DIGESTIBILITY IN BROILERS AT DAY 28 POST HATCH

ABSTRACT

A study was performed varying level of replacement of silkworm meal on soybean

meal on growth performance, apparent metabolizable energy (AME) and apparent

nutrients digestibility in broiler during starter phase. Five diets were calculated on the

basis of iso caloric and iso nitrogenous as 0% (D1), 25% (D2), 50% (D3), 75% (D4)

and 100% (D5) replacement of soybean meal with silkworm meal in broiler starter

ration. A total of 250 day-old broiler chicks were divided into 25 replicate groups

(n=10), with five replicates per diet using a completely randomized design. The

silkworm meal analysis showed higher levels of CP (57.6 ± 1.50%) and crude fat (23.3

± 0.21%). The silkworm meal had higher contents (g/100g of total amino acids) of

lysine (7.52 ± 0.376) and methionine (3.88 ± 0.376), but was slightly deficient in

tryptophan (1.70 ± 0.413). Feed intake, weight gain and dressing percentage were

higher (P < 0.05) in group D4 in comparison with other groups. Moreover, lower (P <

0.05) FCR was observed for D1 and D4 groups than D3 group. AME was higher (P <

0.05) in group D4 and D5 in comparison with other groups. The digestibility of DM,

CP, crude fat, ash and N free extract did not differ (P > 0.05). The Organoleptic quality

parameters were not altered (P > 0.05) due to replacement of soybean meal by

silkworm meal. The findings of the present study revealed that silkworm meal may

effectively be used by replacing 75% soybean meal as low-cost protein ingredient in

broiler ration.

Keywords: Protein source; insect protein; amino acid profile; feed conversion

17

3.1 Introduction

Poultry production mostly depends on plant-seeds based enriched protein of

soybean meal due to its level of digestibility and outstanding amino acid profile (Willis,

2003). Our country Pakistan is limited in production of soybean and most of the

soybean used in poultry ration is imported from other countries. Over a few years the

price of soybean has significantly increased due the rapid growth of poultry industry on

global basis, as well as by the rising demand of soybean in human food industry. The

increasing cost of protein ingredients has provided impetus to the poultry industry by

exploring low-cost, sustainable alternative sources of protein ingredients in poultry

ration (Ramos-Elorduy et al., 2002; Das et al., 2009). Recently, there is an increasing

concern in the use some of the protein-rich insect species as a low-cost protein

ingredient in poultry ration (Van-Hhuis et al., 2013). Insects rapidly grow and fastly

reproduce, had higher feed conversion efficiency, and can be fed on bio-wastes

materials that are efficiently converted into high valuable feed and food resources.

(Collavo et al., 2005) reported that normally about 1000gm of flies can be collected

from 2000gm of waste, but exactly it is more reasonable to harvest insects on

commercial level for a sustainable foundation to mass production of protein for poultry

feed (Veldkamp et al., 2012), and the benefits could be particularly greater, if bio waste

substrate is provided.

Silkworm which can be produced as co-product of the silk industry is a

caterpillar of moth butterfly (Bambyx mori) whose cocoon is used for silk production.

Silkworm meal is an appropriate and suitable source to be used as a livestock feed,

particularly for monogastric animals such as poultry, due to higher content of protein

and better digestibility (Trivedy et al., 2008). Silkworm caterpillar meal is rich in

protein (56.8-63.8%), lipid (19.6-31.5%) and ash (3.3-7.7%) (Loselevich et al., 2004;

Makkar et al., 2014). Apart from these nutrients, their amino acid profiles are

favourably comparable to that of fish meal (Solomon and Yusufu, 2005). However,

according to our knowledge, limited work has been done on the dietary composition of

silkworm meal produced in Pakistan as animal feed. Therefore, the current trial was

planned to quantify the nutritive value of indigenous silkworm meal on the basis of

amino acid and mineral profile to assess its potential as protein ingredient in broiler

feed.

http://journals.cambridge.org.ezproxy.library.wur.nl/action/displayFulltext?type=6&fid=8644389&jid=WPS&volumeId=68&issueId=03&aid=8644388&bodyId=&membershipNumber=&societyETOCSession=&fulltextType=RV&fileId=S0043933912000554#ref044http://journals.cambridge.org.ezproxy.library.wur.nl/action/displayFulltext?type=6&fid=8644389&jid=WPS&volumeId=68&issueId=03&aid=8644388&bodyId=&membershipNumber=&societyETOCSession=&fulltextType=RV&fileId=S0043933912000554#ref029http://journals.cambridge.org.ezproxy.library.wur.nl/action/displayFulltext?type=6&fid=8644389&jid=WPS&volumeId=68&issueId=03&aid=8644388&bodyId=&membershipNumber=&societyETOCSession=&fulltextType=RV&fileId=S0043933912000554#ref010http://www.feedipedia.org/node/4462

18

3.2 Material and methods

3.2.1 Preparation of Silkworm Meal

Dried silkworm pupae with their chitinous covering was obtained from the

Changa Manga Silk Industries (31°05′N latitude 73°58′E longitude) District Kasur,

Pakistan. For silk production, the silkworms were killed in the pupa stage before they

produce enzymes for disruption of silk cocoon. Simultaneously, the cocoon was

removed and used for silk production, and large quantities of spent pupa were produced

as by-product. For the current study, the spent silkworm pupae were obtained from two

consecutive batches at two weeks interval. The spent pupas were dried, crushed

manually and the chitinous matter was removed. The pupa matter was ground and used

for the preparation of experimental diets.

3.2.2 Experimental Design, birds husbandry and diets

A 250 broiler (Ross 308) chicks were randomly separated into 25 replicate

groups. Each replicate group was kept in 10 × 10 feet cages, housed in the same poultry

shed. Five replicates were assigned randomly to each of the five experimental diets

according to a completely randomized design (CRD). The five diets were formulated at

0% (D1), 25% (D2), 50% (D3), 75% (D4) and 100% (D5) substitute of soybean by

silkworm meal in a commercial broiler feed mill. The experimental feeds were pelleted

using experimental pellet machine (Parr Instrument Co, USA) at the University of

Agriculture Peshawar. The ingredient and chemical composition of four diets

formulated on iso-nitrogenous and iso-caloric basis is given in Table 3.1.

Separate feeder and drinker was provided in each cage for provision of water

and feed ad-libitum. Locally adopted vaccination schedule was practiced for

vaccination against prevalent diseases. The trail continued for 28 days and the data of

each replicate regarding the feed intake was documented daily, and at the start of the

experimental trial individual body weight of the birds was recorded and thereafter on

weekly basis. Similarly, the feed conversion efficiency on weekly basis was computed.

At the end of experiment for carcass trait analysis, 3 birds from each replicate were

selected for slaughtering and dressing percentages were recorded.

https://tools.wmflabs.org/geohack/geohack.php?pagename=Changa_Manga&params=31_05_N_73_58_E_region:PK_type:city_source:GNS-enwiki

19

Table 3.1 Ingredient and chemical composition (%) of experimental rations

for broiler at starter phase

Ingredients Diets* (soybean: silkworm)

D1 D2 D3 D4 D5

Corn 54 54 54 54 54

Broken Rice 2.2 2.7 3.2 3.7 4.2

Soybean meal (untoasted) 10 7.5 05 2.5 00

Silk worm meal 0 2 4 6 8

Sunflower meal (fatted) 3.5 3.5 3.5 3.5 3.5

Guar meal (Half toasted) 04 04 04 04 04

Cotton meal (Processed seed) 04 04 04 04 04

Maize gluton meal (30%) 7.3 7.3 7.3 7.3 7.3

Fishmeal 50% 5 5 5 5 5

Molasses 01 01 01 01 01

Rice polish 6 6 6 6 6

Rock phosphate 01 01 01 01 01

Methionine 0.1 0.1 0.1 0.1 0.1

Lysine 0.1 0.1 0.1 0.1 0.1

Lime stone 0.4 0.4 0.4 0.4 0.4

Vitamins Minerals premix 0.1 0.1 0.1 0.1 0.1

Chromic oxide 0.2 0.2 0.2 0.2 0.2

M.E (kcal)/kg 2995 2987 2990 2991 2993

Chemical composition on the basis of ingredients

DM (%) 87.4 87.3 87.2 87.6 88.5

E.E (%) 4.19 4.17 4.18 4.20 4.17

CP (%) 21.2 21.4 21.1 21.3 21.2

CF (%) 3.94 3.92 3.91 3.92 3.93

Crude Ash 5.45 5.43 5.46 5.47 5.48

Phosphorous 0.31 0.30 0.29 0.31 0.30

Ca 0.76 0.77 0.79 0.78 0.77

Cysteine (%)AA 0.32 0.33 0.34 0.32 0.33

Methionine (%) AA 0.48 0.52 0.56 0.59 0.57

Lysine (%) AA 1.39 1.37 1.38 1.39 1.38

Control # (D1), D2- 25%, D3-50%, D4-75%, D5-100% of soybean meal was substituted with silkworm

meal, †SBM, soybean meal; SWM, silkworm meal.

Diet contain: vitamin, vitamin A 12000 IU ; vitamin B1 3 mg; vitamin B2 6 mg; vitamin B6 5 mg;

vitamin B12 0.03 mg; vitamin D3 2400 IU; vitamin E 50 mg; vitamin K3 4 mg; niacin 25 mg; folic acid

1 mg. Mineral, Co 0.2 mg; Mn 80 mg; Cu 5 mg; Se 0.15 mg; calcium-d-pantothenate 10 mg; : Zn 60 mg;

choline chloride 200 mg;

20

3.2.3 Determination of Apparent Metabolizable Energy (AME)

To measure the metabolizable, 5 birds from each replicate group were

transferred to specialized metabolic cages for digestibility analysis. Birds were

individually fed with known quantity of feed. The faeces were collected and

immediately weighed, every next morning, transferred to pre-labelled polythene zip

bags and immediately freeze. Data were collected for 4 consecutive days and samples

were taken at the end of trial for analysis as mentioned by Sultan et al., (2014).The

gross energy of the samples was measured by (Bomb calorimeter- IKA Werke , C7000

& Co. Staufen Germany) and values were calculated by the following formula.

AME = [( )

]

3.2.4 Nutrients digestibility

Nutrients digestibility was determined on day 28 of the experimental trial. For

this purpose, from each group replicate birds (n=5) were chosen and euthanized.

Immediately after euthanization, birds were dissected and samples from ileum

(diverticulum of ileo- cecal junction) and rectum were collected to find out the apparent

total digestibility. For proximate composition, the collected samples were freeze-dried,

ground with the help of 1-mm mesh screen, and analysed in duplicate. The chromic

oxide (Cr2O3) 0.2% was used as indigestible maker to calculate the apparent total

digestibility coefficients. The following formula was used

[

]

Where Cr = chromic oxide

21

3.2.5 Chemical analysis of diets (SBM, soybean meal; SWM, silkworm meal)

The diet and silkworm meal samples were dried in oven at temperature of 60 oC

and milled particles (1 mm) for chemical analysis. The dried, ground samples were

analysed for the contents of dry matter, crude protein, crude fat, and ash according to

the standard procedure of AOAC (2005).The contents of Ca, Mg, Mn, Fe, Cu and Zn

were analysed by atomic absorption spectrophotometer (Buck Scientific 240VGP,

Milan, Italy) according to the procedures of AOAC (1995).The silkworm meal samples

was run for amino acid determination via High Performance Liquid Chromatography

(HPLC) followed the procedures of Cserhati and Forgacs (1999) and Kerese (1984).

3.2.6 Organoleptic study

For organoleptic study different meat cuts from leg, thigh and breast were

cooked up to 82 °C temperature. The cooked samples were evaluated by a board

comprising of members selected from faculty staff and postgraduate students in

accordance with procedure mentioned by Khan et al. (2016). The sample were assessed

on 5-point hedonic scale resulted for colour, taste, flavour, juiciness and tenderness.

3.2.7 Statistical analysis

The outcome of replacing soybean meal with silkworm meal on the production

performance parameters, AME and nutrients digestibility was analysed according to the

standard General Linear Model (GLM) procedure of Statistical Analysis System (SAS,

2009). To compare the differences in case of significant value (P< 0.05) the Tukey

Kramer test were used for pair wise comparison in means. Replicate was used as

experimental unit.

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3.3 Results

3.3.1 Chemical composition of silkworm meal and soybean meal

Data on the biochemical analysis, amino acids and mineral content of silkworm

meal is summarized in Table 2, 3 and 4, respectively. The higher content of CP (57.6%

of DM) in silkworm meal indicates its potential scope as protein ingredient as well as a

rich source of crude fat (23.3%) in poultry feed (Table 3.2).

Table 3.2 Comparative chemical composition of silkworm meal and soybean meal

Composition Silkworm meal Soybean meal

Nutrients Mean ± SE Mean ± SE

Dry matter (DM; g/100 g feed) 92.1 ± 2.23 92.6± 2.11

Crude protein (g/100 g DM) 57.6 ± 1.50 46.1± 1.26

Crude fiber (g/100 g DM) 5.55 ± 0.018 4.05± 1.21

Ash (g/100 g DM) 11.2 ± 1.01 6.41± 1.18

Crude fat (g/100 g DM) 23.3 ± 0.21 1.72± 0.31

Energy (Kcal/100 g DM) 453 ± 8.10 415±7.23

3.3.2 Amino acid profile (g/100g of total) and mineral of silkworm meal

Moreover, almost all the content of important amino acids were present in

silkworm meal which had higher contents (g/100 g total amino acids) of lysine (7.52)

and methionine (3.88), but was slightly deficient in tryptophan (1.70) (Table 3.3). The

silkworm meal was particularly rich in Zn (22.2 mg/kg) and Fe (326 mg/kg) (Table

3.4).

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Table 3.3 Comparative Amino acid profile (g/100g of total amino acids) of

silkworm and soybean meal

Composition Silkworm meal Soybean meal

Amino acids Mean ± SE Mean ± SE

Alanine 5.22 ± 0.255 4.24± 0.231

Arginine 6.31 ± 0.388 2.90 ± 0.212

Aspartic acid 9.59 ± 0.740 11.45 ± 0.614

Cystine 0.97 ± 0.388 0.74± 0.361

Glutamic acid 10.19 ± 1.007 17.91± 1.02

Glycine 4.61 ± 0.376 4.17 ± 0.218

Histidine 3.28 ± 0.425 1.02 ± 0.054

Isoleucine 4.73 ± 0.243 2.07± 0.211

Leucine 7.04 ± 0.303 7.72± 0.213

Lysine 7.52 ± 0.376 2.62± 0.201

Methionine 3.88 ± 0.327 0.52± 0.017

Proline 6.31± 0.510 5.43± 0.209

Phenylalanine 5.58 ± 0.316 2.12± 0.072

Serine 5.22 ± 0.243 4.56± 0.213

Threonine 5.58 ± 0.121 1.66± 0.023

Tyrosine 6.55 ± 0.303 3.11 ± 0.122

Tryptophan 1.70 ± 0.413 0.65± 0.011

Valine 5.70 ± 0.316 2.6± 0.146

Table 3.4 Minerals profile of silkworm and soybean meal

Minerals Mean ± SE Mean ± SE

Calcium (g/kg DM)) 3.8± 2.11 3.7± 1.12

Phosphorus (g/kg DM) 6.2 ± 2.30 5.3± 1.08

Magnesium (g/kg DM) 3.3 ± 2.82 2.4± 0.14

Iron (mg/kg DM) 326 ± 66.1 192±2.42

Manganese (mg/kg DM) 18.2 ±12.12 24.9±1.14

Copper (mg/kg DM) 15.0 ± .01 16.0± 0.42

Zinc (mg/kg DM) 22.2 ± 0.72 48.0± 0.312

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3.3.3 Effect of varying replacement levels of silkworm meal for soybean meal on

performance parameter of broilers

The highest (P < 0.05) feed intake and weight gain was recorded for D4 group

(Table 3.5). The lower (P < 0.05) FCR was observed for D1 and D4 groups in

comparison with D3 group, whereas the FCR for diets D2, and D5 was not affected (P

> 0.05). The D1 group consumed lower amount of feed in comparison with other

groups whereas D2 and D5 groups had lower feed intake than D3 and D4 groups

(P

25

3.7. The digestibility of DM, CP, crude fat, ash and N free extract did not differ

(P>0.05) due to diet composition. The organoleptic properties was not significantly

(P>0.05) affected by the test diets (Table 3.8).

Table 3.6 Effect of varying replacement levels of silkworm meal for soybean

meal on apparent metabolizable energy (AME) of broiler chicks

Diets#(SBM:SWM)† AME

D1 12.7c

D2 12.8bc

D3 12.9b

D4 13.2a

D5 13.3a

SEM 0.080

P-Value ***

abc, means with different letters within column differs at P < 0.05; ***, P < 0.001


meal †SBM, soybean meal; SWM, silkworm meal.

Table 3.7 Apparent nutrient digestibility (%) of different diets fed to broiler

chicks

Digestibility

(%)

Diets# (soybean: silkworm)

SEM Significance# D1

D2 D3 D4 D5

Dry matter 82.5 82.6 82.5 82.7 82.3 2.87 Ns

Crude protein 83.2 81.3 80.2 80.6 79.2.2 3.63 Ns

Ether extract 78.7 79.1 80.5 81.3 80.8 3.59 Ns

Crude fiber 85.3 84.1 85.2 86.1 85.9 1.97 Ns

Ash 49.4 48.5 51.3 52.4 53.3 1.62 Ns

N free extract 90.8 92.7 92.6 93.8 94.1 2.02 Ns

NS, not significant (P> 0.05)



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Table 3.8 Organoleptic quality of broiler meat supplemented silkworm meal

diets

Diets#

(SBM:SWM) †

Taste

(Mean ± SE)

Tenderness

(Mean ± SE)

Juiciness

(Mean ± SE)

Color

(Mean ± SE)

Flavor

(Mean ± SE)

D1 2.31 ± 0.28 2.41 ± 0.23 2.35 ± 0.06 2.45 ± 0.17 2.44 ± 0.55

D2 2.34 ± 0.16 2.37 ± 0.39 2.33 ± 0.15 2.44 ± 0.26 2.52 ± 0.16

D3 2.32 ± 0.27 2.37 ± 0.19 2.32 ± 0.14 2.47 ± 0.38 2.53 ± 0.25

D4 2.41 ± 0.18 2.42 ± 0.13 2.37 ±0.21 2.49 ± 0.22 2.57 ± 0.20

D5 2.43 ± 0.16 2.39 ± 0.23 2.41 ± 0.26 2.42 ± 0.34 2.48 ± 0.31

P-Value Ns ns Ns ns Ns

NS, non-significant

Control #(D1), D2- 25%, D3-50%, D4-75%, D5-100% of soybean meal was substituted with silkworm


3.4 Discussions

The proximate composition indicated that silkworm meal is a rich source of CP

(57.6%) and crude fat (23.3%). Our findings are within the range of values of CP (60.7

± 7.0) and crude fat (25.7 ± 9.0) as reported in a recent Meta-analysis (Makker et al.,

2014). Recently, Ji et al. (2015) stated comparable proximate composition for silkworm

meal, this minor variants in proximate composition might owed the changes in the

substrate of its production, stage of harvesting, and environmental condition and

methods of storage and processing (Ojewola et al. 2005; Ijaiya and Eko, 2009). Unlike

other animal origin feed ingredients, the fat content of silkworm comprises a proportion

of polyunsaturated fatty acids, particularly linolenic acid (18:3n-3), with reported

standards ranges from 11 to 45% of the total fatty acids (Rao, 1994; Usub et al., 2008).

As such supplementation of silkworm meal to poultry birds can potentially improve

their health and production performance, and benefit long-term human health

(Poorghasemi et al., 2013).

The individual amino acid contents of the silkworm meal reported in the present

study is within the range of literature values (Makkar et al., 2014; Ji et al. 2015; Valerie

et al., 2015). The amino acid composition (g/100 g of total amino acids) of the

silkworm meal indicates that amino acids lysine (7.52 vs. 6.11) and methionine (3.88

http://www.feedipedia.org/node/15235

27

vs. 1.40) were higher in silkworm meal than soybean meal (Valerie et al., 2015).

Solomon and Yusufu (2005) and Ji et al. (2015) stated silkworm meal amino acid

profile is almost similar but superior to soybean meal. Moreover, our study showed that

silkworm meal had high proportion (51.3%) of essential amino acids. Specifically, the

silkworm meal contained higher contents (g/100 g of total amino acids) of the essential

amino acids, namely, lysine (7.52), methionine + cystine (4.85), arginine (6.31),

phenylalanine (5.58) and valine (5.70). However, the ratio of leucine and isoleucine

(7.04: 4.73) was lower than that of soybean meal (7.5: 4.6). Leucine and isoleucine

ratio is often connected in amino acid antagonism, therefore, it is necessary to include

in the formulation of poultry ration (Crawshaw, 1994). Moreover, the silkworm meal

was limiting in tryptophan. Roa (1994) also reported that silkworm meal is deficient in

tryptophan and suggested that it would be essential to provide tryptophan along with

the feed that comprises silkworm meal. However, supplementation of the synthetic

amino acids availability, has resolved this issue without costing significantly. The

silkworm also indicated higher gross energy (4530 ± 81.0) and soybean (2250) kcal/kg

according to (NARC, 1994). The Results of present study regarding amino acids profile

inspire the substitution of silkworm meal in the poultry feed.

Summarising available literature data, (Makkar et al., 2014) reported that

excluding silkworm meal, other insects meal are lacking in lysine and methionine,

addition of silkworm meal in the ration can improve both the performance of the

animals and can replace both fish meal and soybean in the diet. The improved

performance of broilers with the replacement of soybean meal with silkworm meal (up

to 75%) could be related to its higher content of essential amino acids, minerals and

energy (Khatun et al. 2003). Fogoonee (1983) argued that the silkworm meal contain

growth prompting factors such as ecdysteroid activity (a hormone involved in protein

synthesis and tissue formation), though this has not been confirmed yet. In this trail

high feed intake, growth potential (body weight gain) dressing percentage and lowest

FCR was observed with 75% replacement of soybean meal by silkworm meal. The

reasons for best FCR with the 75% replacement level may be due to a more optimal

supply content of essential amino acid (predominantly tryptophane), nutrient

digestibility and an higher accumulation rate of protein (Khan et al., 2016). Although

no direct comparisons could be made due to lack of literature data, Fagoonee (1983),

Konwar et al. (2008) and Dutta et al. (2012) stated that replacing 50% of the fish meal

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with silkworm meal in broiler ration supported optimum growth performance and

improved economic return. Moreover, Ijaiya and Eko (2009) reported a higher weight

gain with 75% substitute of silkworm on fish meal. However, feed intake and

production performance reduced with 100 % substitution of silkworm meal on soybean

meal (Ijaiya and Eko 2009; Dutta et al. 2012) which coincide with the findings of our

study. The plausible reasons for the depressed performance of the birds with 100%

replacement of soybean meal with silkworm meal could be the negative impact of high

fat content on intake and the inability of young chicks to utilize the crude fibre inherent

in the exoskeleton (made of chitin) of the silkworm caterpillar (Fagoonee 1983; Makkar

et al., 2014; Valerie et al., 2015).

The findings of our study are also similar to outcomes of Khatun et al. (2008)

stated that higher carcass yield for the dietary groups comprising higher level of fish

meal substituted with silkworm meal. The result of sensory evaluations color, juiciness,

flavour and tenderness showed that the inclusion of silkworm meal did not alter these

properties as related to control diet. In contrast, to our study, Sun et al., (2013) stated

that birds reared on feed containing grasshopper have undesirable effect on meat

sensory quality. The presence of the content of essential amino acids and poly un-

saturated fatty acids in silkworm meal may lead to better sensory scores (Qiao et al,

2016), which were detected in the current study. Our results have showed that the meat

quality of broiler produced with the feeding of silkworm meal is acceptable by the

consumer.

3.5 Conclusions

The finding of this study showed that silkworm is a rich source of CP, crude fat,

and essential amino acids including lysine and methionine. Silkworm meal may be

substituted 75% on soybean meal in broiler feed for better performance and dressing

percentage without affecting meat sensory quality.

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CHAPTER-4

STUDY-II

SILKWORM MEAL AS PROTEIN INGREDIENT IN

BROILER FINISHER RATION

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SILKWORM MEAL AS PROTEIN INGREDIENT IN BROILER

FINISHER RATION

ABSTRACT

This study evaluated the influence of replacing soybean meal with silkworm

meal on production performance, haematology and carcass traits. Five iso nitrogenous

and iso energetic diets were uttered with the step-wise i.e. 0% (D1), 25% (D2), 50%

(D3), 75% (D4) and 100% (D5) replacement of soybean meal with silkworm meal in

commercial broiler finisher ration. A total of 150 broiler (day old: Ross 308) chicks

were randomly separated into 15 replicate groups (n=10), and consequently reared on

five experimental diets according to a completely randomized design. Feed intake,

body weight gain and dressing percentage varied (P < 0.001) due to diet composition,

and the highest (P < 0.05) values were recorded for diet D4. Moreover, numerically

lowest FCR was observed for D1 (control) and D4 diets, whereas the FCR for D2, D3

and D5 diets was not different (P > 0.05). The dressing percentage was not significantly

different (P > 0.05). However analysis of blood parameter and carcass aloof from blood

albumin indicates no significant (P > 0.05) differences among the dietary treatments.

Feed cost per kg turn down

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