THE COMPARISON OF DIFFERENT PELLET FEED ON FEED UTILIZATION, SURVIVAL AND GROWTH OF CLOWN KNIFE FISH (chitala chitala)

CAN THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES THE COMPARISON OF DIFFERENT PELLET FEED ON FEED UTILIZATION, SURVIVAL AND GROWTH OF CLOWN KNIFE FISH Chitala chitala BY VO THI

CAN THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES

THE COMPARISON OF DIFFERENT PELLET FEED ON FEED UTILIZATION, SURVIVAL AND GROWTH OF

CLOWN KNIFE FISH (Chitala chitala)

BY

VO THI Y THU

A thesis submitted in partial fulfillment of the requirements for

the degree of Bachelor of science in Aquaculture

Can Tho, January, 16th 2013

CAN THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES

THE COMPARISON OF DIFFERENT PELLET FEED ON FEED UTILIZATION, SURVIVAL AND GROWTH OF

CLOWN KNIFE FISH (Chitala chitala)

BY

VO THI Y THU

A thesis submitted in partial fulfillment of the requirements for

the degree of Bachelor of science in Aquaculture

Supervisor

Dr LAM MY LAN

Can Tho, January, 16th 2013

Many thanks to Mr Nguyen Hong Quyet Thang, Mss Tran Le Cam Tu and

my friends for their help, preparation of experimental system, sampling, feeding… and analysis data

VO THI Y THU

ABSTRACT

A study on the comparison of different pellet feed on survival rate, growth

and feed utilization of Clown knife fish (Chitala chitala) was set up at College of

Aquaculture and Fishery, Can Tho University

The mean initial weight of fish was 60±5 g/ind Density were stocked 15 ind Into 250L tank Four diet treatments were conducted with three different types

of pellet and the control treatment using trash fish After 8 weeks, the survival rate

of four treatments fluctuated from 75.6 - 97.8% and were significant difference among treatments (p<0.05) The weight of fish in treatments using different pellets was increase during experiment, especially; the survival rate of fish in treatment 2 was highest (69.5±0.94 g/fish) as compared with treatment 1 and 3 The daily weight gain of fish in treatment using trash fish (1496.1±6.7 mg/fish/day) was higher than other treatments and there was significant difference with others (p < 0.05) Besides that, the specific growth rate of control treatment (1551.39±4.61) was also highest after 8 weeks and treatment 2 and 3 were significant difference with treatment 1 and the control ( p<0.05) The food conversion ratio (FCR) of treatment 3 was lowest and was significant difference with treatment 1, 2 and the control (p<0.05) The PER of treatments fluctuated from 0.41% to 0.94%; there was lowest in control treatment The protein content in flesh fish changed 4.24% to 5.14% and there were significantly different between dietary treatments and the control treatment

TABLE OF CONTENT

Acknowledgment i

Abstract ii

Table of content iii

List of tables v

List of figures vi

List of abbreviations vii

Chapter I: INTRODUCTION 1

1.1 Background 1

1.2 Objectives of the study 2

1.3 Research activities 2

Chapter II: LITERATURE REVIEW 3

2.1 Biological characteristic 3

2.1.1 Classification and morphology 3

2.1.2 Distribution characteristics 3

2.2 Growth characteristics 5

2.3 Nutrition characteristics 5

2.4 Study of commercial feed utilization for clown knife fish culture 6

Chapter III: RESEARCH METHODOLOGY 8

3.1 Selection site 8

3.2 Materials 8

3.3 Methodology 9

3.3.1 Experimental management 10

3.3.2 Data analyses 11

3.3.2.1 Evaluations of growth parameters 11

3.3.2.2 Survival rate 11

3.3.2.3 Feed utilization 11

3.3.2.4 Proximate composition of feed and fish 12

3.3.2.5 Data analysis 12

Chapter IV: RESULT AND DISCUSSION 13

4.1 Water quality in experimental in tanks 13

4.1.1 Water temperature 13

4.1.2 pH in experimental 13

4.1.3 DO, ammonium, nitrite 14

4.2 Survival rate 15

4.3.1 The weight gain of fish in experimental 17

4.3.2 Daily weight gain 18

4.3.3 Specific growth rate 19

4.3.4 Food conversion ratio, food intake, protein efficiency ratio 21

4.4 Proximate composition of the experimental diets and fish 23

4.4.1 Proximate composition of experimental diets 23

4.4.2 Proximate composition of fish 24

4.5 Feed cost of diets 25

Chapter V: CONCLUSION AND RECOMMENDATION 27

5.1 Conclusion 27

5.2 Recommendation 27

REFERENCES 28

APPENDICES 32

Appendix 1 32

Appendix 2 34

Appendix 3 35

Appendix 4 37

Appendix 5 37

Appendix 6 38

Appendix 7 39

Appendix 8 40

Appendix 9 41

Appendix 10 42

Appendix 11 43

LIST OF TABLES

Table 3.1 The nutrition composition in commercial feed 7

Table 4.1 Water temperature in experimental tanks 13

Table 4.2 pH in experimental tanks 14

Table 4.3 The fluctuation of DO, ammonium, nitrite 15

Table 4.4 Survival rate of Chitala chitala after 57 days 15

Table 4.5 Weight gain of fish during culture periods 17

Table 4.6 The DWG of treatments in experiment 18

Table 4.7 The specific growth rate of treatments in experiment 19

Table 4.8 FCR, FCE, FI and FER of treatment during culture periods 21

Table 4.9 The proximate composition of the experimental diets 23

Table 4.10 The proximate composition of initial and final fish 23

Table 4.11 The cost for feed of diets 25

LIST OF FIGURES

Figure 3.1 Trash fish 9 Figure 3.2 The composite tank system for experiment 10

LIST OF ABBREVIATIONS

DWG Daily Weight Gain

SGR Specific Growth Ratio

FCR Feed Conversion Rate

FCE Feed Conversion Efficiency

PER Protein Efficiency Ratio

T4 Treatment using trash fish

TAN Total ammonium nitrogen

CAF College of Aquaculture and Fisheries CTU Can Tho university

Chapter I INTRODUCTION

1.1 Background

The Clown knife fish (Chitala chitala) is distributed in some parts of

Southeast Asia, especially in Vietnam Presently, this species has been cultured more in many areas in the Mekong delta because this fish has high value in local market, good meat quality, high price, and growth fast However, in the world, this fish species has not yet studied generous There was some researches on morphology, classification and distribution In Vietnam, some studies were proclaimed about classification of knife fish such as: Truong Thu Khoa and Tran

Thi Thu Huong (1993), fingerling production and feed production for Notopterus

notopterus culture (Tran Ngoc Nguyen, 2000; Le Ngoc Dien, 2004) Although the

knife fish (Notopterus notopterus) has studied and has cultured but this species was small size, most farmers changed to clown knife fish (Chitala chitala) culture

because of large size and fast growth

Presently, most of fingerling production farms fed their fish by fresh foods, worms for fry However, farmers were got more difficulties, especially, they cannot initiative about natural food (depend on the seasons such as flooding season have more different fish species) Using natural food for nursing from fry to fingerling should be replaced by commercial feed to reduce production cost, disease infectious from the live food and initiative of feed for development of clown knife fish farming in pond Therefore, the study on the comparison of pellet feed on survival,

growth of clown knife fish (Chitala chitala) fingerling was carried

1.2 Objective of the study

To find out the suitable feed with optimal feed utilization, survival and

growth for Chitala chitala culture in order apply to the farming practice of this

species in the Mekong delta

1.3- Research activities

- To determine the effect of different types of pellet feed on survival rate and

growth of Chitala chitala

- To determine the effect of using pellet on feed utilization of clown knife fish

- To determine the proximate composition of feed and fish

Chapter II LITERATURE REVIEW

2.1 Biological characteristics

2.1.1 Classification and morphology

According to FishBase (2010), the classification of clown knife fish is:

Kingdom: Animalia

Phylum: Chordata

Class: Actinopterygii

Order: Osteoglossiformes Family: Notopteridae

The clown knife fish is fresh water species Fish can live in water body with low oxygen and pH In the natural condition, this fish lives in the middle or lower reaches of a river or marshes During the day, knife fish usually hides in aquatic plant and dark areas At night, the fish has more activities, catch food, swimming slowly, the dorsal anal operates continuously as waves, fish like live areas with more aquatic plant, pH about 6.5-7.0 (Duong Nhut Long, 2003)

2.2 Growth characteristics

In the natural, the clown knife fish could live about 8-10 years, and could get the size 80 cm in length, and 8-10 kg in weight (Nguyen Chung, 2006) The temperature ranged from 25-32ºC and it was suitable for clown knife fish growth

(Truong Quoc Phu, 2006) In addition, Chitala chitala grew 8-9 cm of length after

4 weeks of nursing (Doan Khac Bo, 2008)

Comparing with other species in the same family of fish, the clown knife fish

grew faster than knife fish (Notopterus notopterus) After one year, the fish was about 30-40 cm in length and weight about 800-1200 g/ind In the pond, Chitala

chitala has size about 500-600 g/ind after 6 months (Duong Nhut Long, 2003)

The stocking density of fry at 100 ind/m² was the best for nursing The fry of this species also feed phytoplankton or zooplankton when the yolk sac was still Therefore, this was so a basic for determination the time when needs supply feed right time that could improve survival rate and growth rate for the nursing (Heming and Budington, 1988) In recent years, clown knife fish was cultured more in many provinces in the Mekong delta The clown knife fishes were stocked with different density and stocking density with 7-15 ind/ m² was better for culture

According to Le Ngoc Dien (2004), the knife fish (Notopterus notopterus )

nursing and growing at the different stocking densities, the daily weight gain (0.23g/day) and survival rate (90.03%) of knife fish at 10 fingerling/m² were higher than stocking density of 20 fingerling/m² (0.16 g/day and 55.20% )

2.3 Nutritional characteristics

According to Heming and Budington (1988), the knife fish is an canivorous species After hatching about 3-4 days, this species fed with small food size including: commercial feed with small size, phytoplankton, zooplankton, soybean,

etc The digestive system of Chitala chitala has including: mouth, gullet, stomach,

gut The ratio between gut length and body length of 30 old days was Li < 1 (Tran

Thi Thanh Hien et al., 2007) The clown knife fish ate fresh foods, insects,

crustaceans, artificial feed (Le Ngoc Dien, 2004) In addition, the stomach of knife

fish contains 25.09% of crustaceans and 17.41% of trash fish (Hossain et al., 1990)

More than 20 days old, clown knife fish could feed commercial feed such as sinking

or floating feed and survival rate of fish to 30 old days was not stable, especially using commercial feed for hatching The survival rate of fish was 74 % (Tran Thi

Thanh Hien, 2008; Nguyen Trung et al., 2000)

Several studies on body structure of fish have been conducted as: mouth site, teeth and size of mouth which can know clearly about natural food types and catching the bail characteristic of fish (Pillar, 1952- cited by Pham Thanh Liem and Tran Dac Dinh, 2004)

The growth rate was important factor that could determine nutrient requirement of aquatic animals The fish would grow fast if they ate enough about quantity and quality of food and suitable food types Conversely, if fishes ate unsuitable food types, fishes would grow slowly or death (Tacon, 1990)

The clown knife fish can attack others fish when they feel hungry In addition, the fish was also shocked by the fluctuation of water quality parameters or changing food suddenly that they did not consume or could be infected by disease to death Therefore, during culture, environmental condition and food should be not change suddenly; the fish should be weaned with new food and feed punctually (Nguyen Chung, 2006)

The growth and survival of early stage of Chitala chitala were studied with

several different type diets as red worm (turbifex) and fresh fish eggs, zooplankton

in both culture systems (nylon hapa and tank) Fishes accepted all types of diets in

experiment rearing with high survival rate (Sarkar et al., 2006)

2.4 Study of commercial feed utilization for clown knife fish culture and other species

Using fresh food for nursing from fry was got difficulties due to fresh food contains pathogen and couldnot initiative food source Therefore, the period time determination of commercial feed utilization and artificial feed types was studied more for most of fish species There were some researches and results in fry stage of

many fish species including: Clarias gariepinus, Coregonus sp., Cyprinus carpio ( Appelbaum et al., 1988; Bergot et al., 1986; Charlon et al., 1986) but some

researches also limited with carnivorous fish species

According to Preson – le Ruyet et al (1995), Watanable and Kiron (1994)

researched on using commercial feed for fry stage was early that was cause of low survival rate and growth slowly Presently, this stage of fish has not yet stomach and digestive enzyme or artificial fish did not stimulate fish catching or decreasing

amount of feed intake of fish (Muran et al., 1990; Person-le Ruyet et al., 1993)

The clown knife fish fry usually ate natural food such as phytoplankton,

zoobenthos after using all yolk sac (Tran Ngoc Nguyen et al., 2000) The fishes

grew more than 20 old days, they could feed artificial feed However, the natural food could not lack in hatching and culturing, especially it was important for first time of using artificial feed Survival rate of clown knife fish fingerlings was higher when using artificial feed (74%), formulated feed combined with red worm (82.7%- 91.3%) (Tran Thi Thanh Hien and Nguyen Huong Thuy, 2008) According to the

study on Micropterus dolomieui (Ehrlich et al., 1989), Clarias macrocephalus (Fermin et al., 1991) were carried out that using a combination of 50% trash fish

and 50% pellet feed for nursing was efficiency than using all commercial feed Survival rate and growth of fish were improved when apply those feed

According to Phan Quoc Thu (2009), the growth of clown knife fish rearing

in the hapas feeding fed 50% trash fish and 50% commercial feed was lower than feeding commercial feed of fish However, survival rate of fish in both experiment was not significant different Amount of trash fish was replaced increasing by commercial feed that affected to the growth of fish

The clown knife was hatched and reared by using trash fish Besides that,

Chitala chitala was also accepted pellet feed with different protein levels such as

25-30% protein for hatching, 20-25% protein for culturing and their daily weight gain was also increase, but using homemade feed as 50% pellet feed with 20% protein and 50% trash fish was used and the daily weight gain was increase better the growth and survival (Le Ngoc Dien, 2004)

The survival rate of clown knife fish was not affected by different protein levels The daily weight gain of fish fed protein levels from 40.3% to 65.7% was from 0.53 to 0.9 g/day, respectively (Ho Duc Toan, 2011) According Le Thi Thuy Trang (2012), the growth of fish fed high protein level (40%) was 17.68 and increased higher than fish fed low protein level (30%) In addition, the survival rate

of fish was not affected by using commercial feed with different protein levels

Chapter III RESEARCH METHODOLOGY

Feed source:

 Different types of commercial pellet feed were used (homemade feed with 40% protein, Cavang (goldfish), Tongwei) which were the similar protein (40% protein)

 One type of feed was trash fish (marine fish)

Table 3.1 Proximate composition of dietary (dry matter)

 12 composite tanks: 250 L each

 Water resource: tap water, recirculation system

 Test kit: DO, TAN, pH and nitrite

 Chemical: Chlorine, salt

 Electric balance (1 g), aerator system, net, tray

3.3 Experimental design

Experiment was conducted in composite tank system Each tank had aerated and supplied water continuously Before experiment was carried out, tank system had been treated by 1 kg of salt and wash by clean water

Experiment was carried out in 8 weeks Fish was sampled randomly with 15 individuals / tank The experiment had 3 treatments of different pelltes(Tongwei, Cavang (goldfish) and homemade with 40% protein were called A, B and C diet) and the control treatment (marine fish) with three replications for each treatment

Figure 3.1 Marine fish

Figure 3.2: The composite tank system (250L/tank) for experiment

3.3.1 Experimental management

During experiment, the experimental fish were fed two times per day (7am and 4pm) The fish activity was daily observed such as swimming, feeding Siphon bottom of tank and remove wastes out and uneatten feed with 2 days/time Each tank had aerator continuously

3.3.2 Data collection and analysis

Fish was cultured in tanks for 8 weeks Water quality parameters as pH, nitrite, TAN and DO were measured as follows:

 Nitrite: be measured biweekly at 7 am by using sera test kit

 Total Ammonium nitrogen: be measured biweekly at 7 am by using sera test kit

 pH: be measured biweekly at 7 am by pH meter

 Temperature: be measured biweekly at 7 am by dipping a mercury thermometer into the water surface

3.3.2.1 Evaluation of growth parameters Fish growth performance was determined biweekly by all weighing

-Specific growth rate (%/day)

SGR= (lnWf – lnWi)*100/t

Wi: mean initial fish weight (g)

Wf: mean final fish weight (g)

-Daily weight gain (g/day)

DWG= (Wf – Wi)/ t

Wi: mean initial fish weight (g)

Wf: mean final fish weight (g) t: cultured period (day)

3.3.2.2 Survival rate (%)

During the experiment, the mortality rate of fish was recorded

Number of fish at stocking SR=100*

Number of fish at harvest

3.3.2.3 Feed utilization

- FCR: food conversion ratio

FCR= dry food fed/ wet weight gain

- FCE: food conversion efficiency

FCE = 1/ FCR

- PER: protein efficiency ratio

PER= weight gain/ protein intake Protein intake = feed intake * crude protein (%) in feed

- FI: feed intake (g/ fish/ day)

FI= dry feed intake/ fish/ t

t: cultured period (day)

3.3.2.2 Proximate composition feed and fish

The flesh was collected and 2 times: at stocking and harvesting fish for analysis The proximate analysis of experimental diet and fish were analyzed by the following methods:

+ Moisture: the sample was dried in the oven at 105ºC for 4 hours and weighing the sample

+ Crude protein (CP): analyzed by using the Kjeldahl method

+ Crude lipid (CL): analyzed by Soxhlet method with Chloroform solution The lipid in the sample was extracted perfectly from chloroform washing

+ Ash: was determined by weighing a dry subsample in a preweighed procelain crucible for 8 hours at 600ºC

3.3.2.5 Data analysis

Mean value of treatments were statistically analyzed using one way ANOVA through SPSS version 16.0 The mean among the treatments were significantly compared at p< 0.05 by Duncan Test

CHAPTER IV RESULTS AND DISCUSSION

4.1 Water quality in experimental tanks

4.1.1 Water temperature

Table 4.1 The fluctuation of water temperature (ºC) during culture periods

The temperature in the experimental tanks fluctuated from 27ºC to 29ºC At the first and second sampling time, the temperature was 29ºC, and higher than that

at third and fourth sampling time The water temperature is one of the important factors that affects on the growth According to Truong Quoc Phu (2006), the water temperature from 20ºC to 30ºC was suitable for optimal growth of fish

4.1.2 pH in the experiment

Table 4.2 The fluctuation of pH in experimental tanks

pH in the water fluctuated from 7.5 to 8.5 At the first sampling time, water

pH was 7.5 From the second sampling time to the end of experiment period pH increased to 8.5 pH fluctuation went up due to break up of feed or feces of fish into water However, the water pH were in suitable ranges (6.5- 9) for optimal growth of knife fish (Truong Quoc Phu, 2006) Comparison with the study on different protein

levels in homemade feed for Chitala chitala culture conducted by Ho Duc Toan

(2011), pH in experimental tanks was 7.5 and lower than the present study

4.1.3 Dissolved oxygen, total ammonium nitrogen and nitrite (mg/L)

During experimental periods, the tank system was supplied enough oxygen

by aeration Therefore, the DO in all tanks did not fluctuate, and was 5 mg/L According to Truong Quoc Phu (2006), the optimal of dissolved oxygen for fish growth were 5 mg/L DO in during experiment periods was maintained higer than 5mg/L for growth of clown knife fish

Nitrite content was not much fluctuated among treatments The nitrite values varied from 0.05 mg/L to 1 mg/L and was suitable for fish growth According to

Boyd (1990), TAN concentration was 0.6 to 2mg/L that was toxic for aquatic animal Therefore, the result of TAN was in suitable range for optimal fish growth

Table 4.3 The fluctuation of TAN (mg/L) and NO2- (mg/L) at different sampling time

4.2 Survival rate

Table 4.4 Survival rate of clown knife fish after 8 weeks

The survival rate of all treatments fluctuated from 75.6% to 97.8% (Table

4.4) During the experiment period, the survival rate of treatment 4 was the highest and significant difference with the treatment 1 and 2 (p<0.05) The survival rate of fish in the treatment 2 was the lowest The survival of treatment 3 was no significant difference with treatment 4 However, there was significant difference with the treatment 2 Besides, the survival of the treatment 1 was no significant difference (p

< 0.05) with the treatment 2 and 3 but it was significant difference with the treatment 4 (p < 0.05)

In fact, disease factor was also one of causes that affected to fluctuation of survival In the treatment 1 and 2, survival went down 75.6% and 84.4% (table 4.4) due to parasites and fungus occurred and infected to experimental fish The survival rate of treatments using different commercial feed of the present study was lower than the result of Le Thi Thuy Trang (2012), the survival rate of Clown knife fish fed different protein content was range 96,7% to 100% Furthermore, the study on determination protein requirement of knife fish with size 2-3 g/ fish was 68,9 - 84,4% of survival rate (Nguyen Thi Thuy Kieu, 2011) In addition, the survival rate

of Chitala chitala fed different pellets feed was lower than the result of Ho Duc

Toan (2011), the control fed trash fish was 98.3% However, those results was higher than the study on clownknife culture in hapa of La Anh Nguyet (2008), the survival of fish was 70% in treatment using trash fish and 77% in treatment fed commercial feed

4.3 Fish growth performance

4.3.1The weight gain of fish

Table 4.5 The mean weight of fish (g/fish) during cultured periods

The mean weight of clown knife fish was showed in Table 4.5 During 15

days, 30 days and 45 days, the weight gain of fish in the treatments using different pellet feed were no significant difference (p>0.05); but they were significant difference with the control treatment (p<0.05) Overall, weight of fish in the control treatment was higher than others due to the control used trash fish with high protein

In final harvesting, the weight of fish was 145.7 g/fish in the control and were significantly biggest (p<0.05) Comparison with the result of Phan Quoc Thu (2009), the initial weight of fish was 4.33 g/fish and final weight was 74.7±5.71g in the treatment using 100% trash fish Furthermore, the growth of fish in treatments was increase, respectively Many research showed that the growth rate of fish tended to decrease when they fed feed with higher than 40% of protein level; the

study of Nguyen Tan Dinh (2000) on Pangasius kunyit indicated that the weight of

fish was highest in 30% of protein level and deceased in 40% protein level

Mystus nemurus increased when using protein from 27%-42%, however, growth

rate of fish was decreased while they fed higher than 42% of protein level

4.3.2 Daily weight gain

Table 4.6 The daily weight gain (mg/day) of fish in four treatments

treatment first time

At second and third sampling, there was significant difference between treatment using different pellets feed and control treatment (p<0.05) The daily weight gain (DWG) of control treatment was the highest (1,260±13.8 mg/day) In addition, DWG of treatment 3 was increased and higher than treatment 2 and treatment 1 at the second sampling time, and was significant difference (p< 0.05) The daily weight gain of diets treatment and control treatment were higher than first sampling (Table 4.5) At the third sampling, the concentration of NO2-, NH3+ in

water were higher than first time due to the feces, uneatten feed more in water In of case, disease infection occurred in culture system as fungus, parasite that affected to the health of fish and growth rate of fish in each sampling

The DWG of Chitala chitala in the control treatment after 8 weeks culture

was different from treatments using pellet feed (p<0.05) Besides that, among using commercial feed treatments were not significantly different due to equivalent protein level La Anh Nguyet (2008) showed that the daily weight gain of trash fish treatment was 1.4 ± 0.12 g/day after 2 months This result was similar to treatment

4, and the result of daily weight gain of treatment 1, 2 and 3 (Table 4.6) were lower than daily weight gain of using commercial feed treatment for clown knife fish culture (0.6 ± 0.3 g/day) of research of La Anh Nguyet (2008) According to the result of Phan Quoc Thu (2009), the DWG of clown knife fish feeding trash fish treatment was 3.23 ± 0.01 g/day and pellet feed treatment was 1.81 g/day and higher than all treatments of this present study Compared with study of Le Thi Thuy

Trang (2012) on Chitala chitala, the fish fed with 40% protein, DWG was

0,32±0,04 g/day and higher than the result of the present study