study on the replacement of fish meal protein by soybean meal protein in feed for clown knifefish (chitala chitala) juvenile

CAN THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES STUDY ON THE REPLACEMENT OF FISH MEAL PROTEIN BY SOYBEAN MEAL PROTEIN IN FEED FOR CLOWN KNIFEFISH Chitala chitala JUVENILE BY

CAN THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES

STUDY ON THE REPLACEMENT OF FISH MEAL PROTEIN

BY SOYBEAN MEAL PROTEIN IN FEED FOR

CLOWN KNIFEFISH (Chitala chitala) JUVENILE

BY

NGUYEN NHAT MINH QUANG

A thesis submitted in partial fulfillment of the requirements for

the degree of Bachelor Science in Aquaculture

Can Tho, January 16th, 2013

CAN THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES

STUDY ON THE REPLACEMENT OF FISH MEAL PROTEIN

BY SOYBEAN MEAL PROTEIN IN FEED FOR

CLOWN KNIFEFISH (Chitala chitala) JUVENILE

BY

NGUYEN NHAT MINH QUANG

A thesis submitted in partial fulfillment of the requirements for

the degree of Bachelor Science in Aquaculture

Supervisors Ass Prof Dr TRAN THI THANH HIEN

MSc TRAN LE CAM TU

Can Tho, January 16th, 2013

Acknowledgements

First of all, I wish to express my deep appreciation to Ass Prof Dr Tran Thi Thanh Hien and MSc Tran Le Cam Tu for their constant guidance They have enthusiastically instructed and facilitated me to finish my thesis recently I honestly thank to Board directors of CAF and teaching staffs of AAP supported the knowledge to me in 4.5 years

I also want to show my deep gratitude to my advisor Dr Pham Minh Duc for his constant support and encouragement during learning time in College of Aquaculture and Fisheries

I want to thank my sister Nguyen Thi Linh Dan and my classmates in Advance Aquaculture Program Class so much for encouraging and helping me in last time

Finally, I want to thank Advance Aquaculture Program for supporting expenses to complete my thesis

Abstract

The study was conducted in plastic tanks (80 L /tank) to evaluate the effect of replacement of fish meal protein by soybean meal protein in diets of clown knife fish juveniles Triplicate groups of fish with the initial weight 4.57 g ± 0.07 were fed with 5 diets in which 0%, 15%, 30%, 45%, 60% fish meal protein were replaced by soybean meal protein After 9 weeks, the results showed that fishmeal protein replacement levels were not significantly different on survival rate (SR) (p>0.05) Basing on the significant difference of fish weight gain (WG), food conversion ratio (FCR), protein efficiency ratio (PER); the results were indicated up to 45 % fish meal protein replaced by soybean meal protein without causing reduction on growth index Replacing 60% of fish meal protein

by soybean meal protein resulted in lower growth value than the control diet (p<0.05) Soybean meal contained little essential amino acids (Methionine and Lysine); however, replacing fish meal protein by soybean meal protein without supplement essential amino acids did not affect to the growth and protein efficiency ratio of experimental fish

Table of contents

CHAPTER I: INTRODUCTION 1

1.1Background of the study 1

1.2 Objectives of the study 1

1.3 Contents of the study 2

CHAPTER II: LITERATURE REVIEW 3

2.1 Clown knife fish 3

2.1.1 Biological characteristics 3

2.1.3 Growth characteristics 4

2.1.4 Protein and amino acid requirement of fish 4

2.2 Popular ingredients of feed in aquaculture 5

2.2.1 Fish meal 5

2.2.2 Soybean meal 5

2.3 Role of protein in diet 8

2.4 Replacement fish meal protein by other protein sources in fish diet 9

CHAPTER III: METHODOLOGY 10

3.1 Experimental design 10

3.2 Experimental fish 10

3.3 Experimental diet 11

3.3.1 Formula of diet 11

3.3.2 Proximate compositions of diet 11

3.3.3 Contents of essential amino acids in experimental diets 12

3.4 Experimental system and management 12

3.5 Data collection and analysis 12

3.6 Statistic analysis 13

CHAPTER IV: RESULTS AND DISCUSSIONS 14

4.1 Environmental conditions 14

4.2 Amino acid utilization 14

4.3 Survival rate 16

4.4 Growth performances 16

4.5 Feed utilization 18

CHAPTER V: CONCLUSIONS AND RECOMMENDATIONS 22

5.1 Conclusions 22

5.2 Recommendations 22

References 23

Appendices 28

List of tables

Table 2.1: Proximate compositions of soybean meal 6

Table 2.2: Essential amino acids in soybean meal 7

Table 3.1: Composition of five experimental diets 11

Table 3.2: Proximate compositions of experimental diet 11

Table 4.1: Experimental parameters 14

Table 4.2: Methionine and Lysine in diet and fish 15

Table 4.3: Growth performances of experimental fish 17

Table 4.4: Feed utilization of experimental fish 18

List of figures

Figure 4.1: Survival rate of experimental fish after 9 weeks 16Figure 4.2 : Food conversion ratio of clown knife fish after 9 weeks 20

List of abbreviations

CHAPTER I INTRODUCTION 1.1 Background of the study

Clown knife fish (Chitala chitala) is one of species which have high economic value

Besides, they can be cultured easily in tropical country Their fillet have the high quality and can be processed many kinds of food Clown knife fish is carnivorous species; they often use the feed which has the origin from animal protein source, such as: insects, crustaceans, zooplankton…

Food is major effect to economic efficiency The cost of feed is usually very high proportion in total costs Moreover, fish meal has a scarce tendency Thus, finding other sources of protein to replace fish meal protein source is necessary and urgent Among the plant protein sources, soybean meal is considered as the source which has very bright prospect when replacing a part or all of fish meal in diet, because it contains high concentration of protein and balances in essential amino acids Besides, the source of soybean meal is stable and the price is suitable as much as possible (Hertrampf and Piedad – Pascual, 2000) Protein in soybean meal replacing by protein in fish meal has been studied for a long time and got many achieved successes in fresh water species as well as marine species which are economically important, such as Tra catfish (Tran Thi Thanh Hien and Le Quoc Phong, 2011), African sharptooth catfish (Fagbenro and

Davies, 2001), rainbow trout (Cheng et al., 2003), tilapia (Agbo, 2008; Tri and David,

2009), black spotted grouper (Le Anh Tuan, 2006), and cobia (Pham Duc Hung and Nguyen Dinh Mao, 2009)

However, there are not any researches on replacement of fish meal protein by soybean

meal protein in feed for clown knife fish (Chitala chitala) Thus, the usability studies of

soybean meal as feed are actually necessary

1.2 Objectives of the study

To determine the appropriate level of replacing fishmeal protein by soybean meal protein

in feed for Chitala chitala to develop feed formulation and reduce the cost of feed for

clown knife fish

1.3 Contents of the study

 Evaluation on the effect of different levels of replacing fishmeal protein by

soybean protein on Chitala growth

 Evaluation on the effect of replacing fishmeal protein by soybean protein on feed efficiency of clown knife fish

CHAPTER II LITERATURE REVIEW

2.1 Clown knife fish

two sides and belly is white There is a row of black dots with white edge circle along to the anal fin in the tail

(Source: http://www.hua.edu.vn/khoa/cnts/index.php?option=com_content&task=view&i d=1685&Itemid=211)

2.1.2 Nutritional characteristics

Clown knife fish is carnivorous fish species feeding on both plants and animals (fish, insects, and shrimp), they are active at night Food for them is earth – worm, bait fish, trash fish, processing feed which were made from agricultural waste sources such as: vegetables, water hyacinth, bran, crabs and snails shred

(Source: http://www.hua.edu.vn/khoa/cnts/index.php?option=com_content&task=view&i d=1685&Itemid=211)

2.1.4 Protein and amino acid requirement of fish

Protein is a key component in the organism and responsible for building the body structure In aquatic animals, protein contents about 60 – 75% of body weight In diet, protein provides amino acids by the digestive process and hydrolysis The amino acids are absorbed into the bloodstream and transferred to the agencies for involving in protein biosynthesis process They serve in developing, reproducing and maintaining the living body Thus, we should provide sufficient protein demand for fish in the process of culturing Protein demand of fish ranges about 25 – 55%, average is 30% Fish are omnivorous or herbivorous species need about 25 – 35% protein in diet The protein

demand of carnivore is higher (40 – 52%) (Le Thanh Hung, 2008)

Protein is formed by amino acids In nature, there are more than 200 amino acids, but only about 20 popular types Among of them, there are 10 amino acids that fish do not have the ability to synthesize, and must absorb them in feed, such as: Methionine,

Arginine, Threonine, Tryptophan, Histidine, Isoleucine, Lysine, Leucine, Valine and Phenylalanine Lysine and Methionine are often missed; they are essential amino acids and need to be available in diet Basing on that, there are many studies on these two amino acids on fresh water, brackish water and brine

reduce the stress response (Michael New et al., 1995)

2.2.2 Soybean meal

Soybean meal is the product remaining after extracting most of the oil from soybeans The oil may be removed by solvent extraction or by an expeller process in which the beans are heated and squeezed The nutrient composition of defatted soybean meal is shown in the table below

Table 2.1: Proximate compositions of soybean meal

(Source: Hertrampf and Piedad-Pascual, 2000) Soybean meal consists of about 44-48% protein, 0.5-5% lipid, 6-6.6% fiber and 10-11% moisture Soybean meal is the vegetable protein source which supplied to aquatic feed All over the world, people produce 85-100 MT soybean meal every year, they contain more than 50% protein source in feeding for aquatic animal and cattle Soybean meal contains most of essential amino acids However, there are imbalanced among of amino acids, because low content of Methionine and Cystine compared with fish demanding The table of essential amino acids in soybean meal will be shown below (Hertrampf and Piedad-Pascual, 2000)

Typical Analysis (Solvent extracted soybean meal)

Total Digestible Nutrients

Net energy – Lactation

7.1% 5.3% 0.2% 0.65% 78% 81.1 Mcal /100lbs

Table 2.2: Essential amino acids in soybean meal

(Source: Hertrampf and Piedad-Pascual, 2000) Soybean is one of the world’s best non-fish sources containing essential omega-3 fatty acids, healthy proteins, and unsaturated fats High-quality soy protein is fed to farmed fish and shellfish, to support their growth and healthy development Comparing with other animal protein source, soybean meal is lack of some essential mineral salt such as phosphorus Phosphorus has low value of digestion, they form in phytic acid; therefore, they cannot be absorbed There were many researches on replacing fishmeal protein with soybean meal protein in aquaculture which show that fish or shrimp development decreased when using absolutely soybean meal protein If we supplied fish oil and essential amino acids, we could replace 75% fish meal with soybean meal When using 100% soybean meal, the development of fish will decrease because the feed is limited in energy, lysine and methionine Thus, in reality, soybean meal has to be mixed with fishmeal or animal protein source in a particular ratio to adding some disadvantages of soybean meal in fish diet (Teresa Banaszkiewicz, 2011)

Essential amino acids (% protein)

2.3 Role of protein in diet

Because protein is the most expensive part of fish diet, it is important to accurately determine the protein requirements for each species and size of cultured fish Proteins are formed by linkages of individual amino acids Although over 200 amino acids occur in nature, only about 20 amino acids are common Of these, 10 are essential (indispensable) amino acids that cannot be synthesized by fish The 10 essential amino acids that must be supplied by in diet are: Methionine, Arginine, Threonine, Tryptophan, Histidine, Isoleucine, Lysine, Leucine, Valine and Phenylalanine Among them, Lysine and Methionine are often the first limiting amino acids Diet which is prepared with plant (soybean meal) protein typically is low in methionine; therefore, extra methionine must

be added to soybean meal to promote optimal growth and health It is important to know and match the protein requirements and the amino acid requirements of each fish species

reared (Steven Craig et al., 2009)

Protein level in aquaculture feed is generally average less than 30% for marine shrimp, 28-32% for catfish, 32-38% for tilapia, 38-42% for hybrid striped bass Protein requirements usually are lower for herbivorous fish (plant eating) and omnivorous fish (plant-animal eaters) than they are for carnivorous (flesh-eating) fish, and are higher for fish reared in high density (recirculating aquaculture) than low density (pond aquaculture) systems In growth-out stage, protein requirements of fish usually decrease Protein requirements also vary with rearing environment, water temperature and water quality, as well as the genetic composition and feeding rates of the fish Protein is used for fish growth if adequate levels of fats and carbohydrates are present in the diet If not,

protein may be used for energy and life support rather than growth (Steven Craig et al.,

2009)

Proteins are composed of carbon (50%), nitrogen (16%), oxygen (21.5%), and hydrogen (6.5%) Fish are capable of using a high protein diet, but as much as 65% of the protein may be lost to the environment Most nitrogen is excreted as ammonia (NH3) by the gills

of fish, and only 10% is lost as solid waste Accelerating eutrophication (nutrient enrichment) of surface water due to excess nitrogen from fish farm effluent is a major water quality concern of fish farmers Effective feeding and waste management practices

are essential to protect downstream water quality (Steven Craig et al., 2009)

2.4 Replacement fish meal protein by other protein sources in fish diet

Firstly, replacement fish meal by other protein sources is essential for developing a sustainable aquaculture If aquaculture wants to be sustainable, farming should be friendly with the environment Make sure that doing aquaculture did not reduce water quality, destroy ecological balance, and cause damage to other natural resources; including habitat for other wild species, water, and biological environment Secondly, total yield of trash fish caught in the wild to make fish meal is severely reduced The fishing vessels must exploit deeper and further offshore This is not only a warning about the shortage of fish meal which provides for producing aquatic food in the future but also

a sign about the imbalanced ecological environment and these fish are also an important food source for other wild fish in the ocean Finally, demand for industrial food intake also increased significantly because of rapid growth of aquaculture production Statistics

in recent years shows that the amount of feed used in aquaculture industry has grown 30% annually Therefore, the supply of fish meal from nature can not satisfy the demand for this

(Source: http://csvtsnt.ning.com/forum/topics/dinh-dong-va-cong-nghesan?groupUrl=ntts 43sg&groupId=2963669%3AGroup%3A7240&id=2963669%3ATopic%3A58847&page

=1#comments)

Meanwhile, soybean meal is considered as a source of vegetable protein which is more likely to replace fish meal in feed production technology We can say that replacing fish meal by soybean meal is very good because of high protein content, cheaper, more stable

in quality and full of essential amino acids Moreover, several studies also showed that soybean meal could replace a large portion of fish meal in feed formulation and application of fisheries for many different species of aquatic animals in the world (Source: http://csvtsnt.ning.com/forum/topics/dinh-dong-va-cong-nghesan?groupUrl=ntts 43sg&groupId=2963669%3AGroup%3A7240&id=2963669%3ATopic%3A58847&page

=1#comments)

CHAPTER III METHODOLOGY 3.1 Experimental design

The experiment was carried out with five treatments The percentage of fishmeal protein was replaced with soybean meal protein from 15-60%, and one control treatment without soybean meal Each treatment was repeated three times The stocking density was 30 individuals /80 L

 Treatment 1: 0% soybean meal protein (control treatment)

 Treatment 2: 15% soybean meal protein

 Treatment 3: 30% soybean meal protein

 Treatment 4: 45% soybean meal protein

 Treatment 5: 60% soybean meal protein

Five treatment diets were formulated to be isonitrogenous 40% in crude protein and 4.5 Kcal/g diet in gross energy The control diet was set with fish meal (FM) as the main protein source In diets 2 to 5, soybean meal (SM) was substituted for an isonitrogenous amount of FM control diet, to achieve 15% (diet 2 - SM 20), 30% (diet 3 - SM 30), 45% (diet 4 - SM 40) and 60% (diet 5 - SM 50) of the FM crude protein

Approximate composition includes dry matter, crude protein, crude lipid, ash, fiber, nitrogen free extract, gross energy Ingredients in diet were fish meal, soybean meal, starch, vitamin, mineral, fish oil and Carboxyl Methyl Cellulose (CMC)

The experimental diets were made in a laboratory mill by blending all of the dry ingredients The extruding temperature exceeded 1000C After extruding, all diets were dried at 600C within 48h and stored in refrigerator to use

The experiment was carried out from June 2012 to December 2012

Location: College of Aquaculture and Fisheries – Can Tho University

3.2 Experimental fish

Experimental fish were bought before carrying out the research so that fish could be familiar with the pellets The average size was about 4 – 5g/individual