Modulation of growth performances, survival, digestive enzyme activities and intestinal microbiota...

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Modulation of growth performances, survival, digestive enzyme activities and intestinal microbiota in common carp

(Cyprinus carpio) larvae using short chain fructooligosaccharide

Journal: Aquaculture Research

Manuscript ID: Draft

Manuscript Type: Original Article

Date Submitted by the Author: n/a

Complete List of Authors: Ehsaghzadeh, Hamid; Young Researchers Club, Lahijan Branch, Islamic Azad University, Lahijan, Iran, Hoseinifar, Seyed Hossein; Gorgan University of Agricultural Sciences and Natural Resources, Fisheries and environment; vahabzadeh, Habib; Lahijan Branch, Islamic Azad University (IAU), Lahijan, Iran, Department of Fisheries,

Keywords: sc-FOS, Prebiotic, digestive enzyme activities, intestinal microbiota, Cyprinus carpio

Aquaculture Research


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Modulation of growth performances, survival, digestive enzyme activities and 1

intestinal microbiota in common carp (Cyprinus carpio) larvae using short 2

chain fructooligosaccharide 3

4

Running title: short chain fructooligosaccharide and carp larvae 5

6

Hamid Ehsaghzadeh a*

, Seyed Hossein Hoseinifar b, Habib Vahabzaheh

c 7

8

* b

Young Researchers Club, Lahijan Branch, Islamic Azad University, Lahijan, Iran 9

a Department of Fisheries, Faculty of Fisheries and Environmental Sciences, Gorgan University 10

of Agricultural Sciences and Natural Resources, Gorgan, Iran 11

c Department of Fisheries, Lahijan Branch, Islamic Azad University (IAU), Lahijan, Iran 12

d Culture and propagation department, International Sturgeon Research Institute, Rasht, Iran 13

14

*Author for correspondence: 15

H. Eshaghzadeh, Department of Fisheries, Young Researchers Club, Lahijan Branch, Islamic 16

Azad University, Lahijan, Iran 17

E-mail: [email protected] 18

19

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Abstract 20

This study investigates the effects of inclusion of low levels of dietary short chain 21

fructooligosacchairde (sc-FOS) on physiological response and intestinal microbiota of carp 22

(Cyprinus carpio) larvae. After acclimation, fish (550 ± 20 mg) were allocated into 9 tanks (40 23

fish per tank) and triplicate groups were fed a control diet (0%) or diets containing 0.5% and 1% 24

sc-FOS for 7 weeks. At the end of the experiment, the growth performance parameters (final 25

weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR), and condition 26

factor (CF), survival rate as well as digestive enzyme activities (amylase, lipase and protease), 27

total viable counts of heterotrophic aerobic bacteria (TVC) and lactic acid bacteria (LAB) level 28

in intestinal microbiota were measured. Our results revealed no significant (P > 0.05) effects of 29

sc-FOS on growth performance and TVC when compared to the control group. However, 30

administration of low levels of dietary sc-FOS significantly increased digestive enzyme activities 31

(lipase and amylase) and lactic acid bacteria (LAB) levels (P < 0.05). Also, survival rate was 32

significantly elevated in sc-FOS fed carp. This study inspires further research on various aspects 33

of sc-FOS administration in carp larvae culture. 34

Key word: sc-FOS, prebiotic, growth, intestinal microbiota, digestive enzyme activities, 35

Cyprinus carpio 36

Introduction 37

Administration of antibiotics in aquaculture, especially in intensive systems was an important 38

concern and resulted in ban or restriction of antibiotic usage in many countries (Cabello, 2004). 39

To resolve this issue, several researches performed to investigate dietary supplements as an 40

alternative of antibiotics in aquaculture (Merrifield, Dimitroglou, Foey, Davies, Baker, Bøgwald, 41

Castex & Ringø 2010; Ringø, Dimitroglou, Hoseinifar & Davies 2014). Prebiotics, non-42

digestible dietary ingredient which can beneficially affect intestinal microbiota, has the potential 43

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to be used as an alternative of antibiotics. It has been reported that prebiotics can increase growth 44

performance and improve health status of fish and shellfish through modulation of 45

gastrointestinal tract microbiom toward potentially beneficial communities like Lactic Acid 46

Bacteria (LAB) (Gatesoupe, 2008; Merrifield, Balcázar, Daniels, Zhou, Carnevali, Sun, 47

Hoseinifar & Ringø 2014; Ringø et al., 2014). LAB have been considered as beneficial residents 48

of the fish intestinal ecosystem and frequently isolated from the gut of several fish species 49

(Dimitroglou, Merrifield, Carnevali, Picchietti, Avella, Daniels, Guroy & Davies 2011). 50

Although they are not generally considered to be the dominant species, but elevation of their 51

levels has been reported following administration of prebiotics (Ringø, Olsen, Gifstad, Dalmo, 52

Amlund, Hemre & Bakke 2010). 53

Inulin and fructooligosaccharide are the two common prebiotics extracted from the chicory roots 54

(Niness, 1999). The beneficial effects of dietary inulin and FOS have been reported on growth 55

performance, digestive enzyme activities and health status of several fish and shellfish species 56

(Ringø et al., 2010; Merrifield et al., 2010; Ringø et al., 2014). However, differences in the 57

dosage, species and life stages in different experiments prevent to identify their beneficial and 58

exact effects on the purpose organism (Denev, Staykov, Moutafchieva & Beev 2009). Despite, 59

numerous studies on administration of prebiotics in aquaculture, limited information is available 60

on the effects of prebiotics on growth performance, carcass composition and digestive enzymes 61

activities in early life stages of common carp (Yanbo & Zirong, 2006; Ringø et al., 2010; Ringø 62

et al., 2014). Thus, the present study was performed to investigate the effects of sc-FOS on the 63

growth performance, survival rate, digestive enzymes activities and intestinal microbiota of 64

common carp larvae. 65

Material and Methods 66

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Prebiotic 67

Three levels of sc-FOS (TEREOS-SYRAL, Marckolsheim, France); 0% (control), 0.5% and 1% 68

were added to a basal formulated diet (Table 1) for preparation of experimental diets. The 69

ingredients were blended thoroughly in a mixer and pelleted using a meat grinder equipped with 70

a 2-mm die. The pelleted diets were air-dried and stored in plastic bag at -4oC until further use. 71

Experimental diet 72

Three levels of sc-FOS; 0% (control), 0.5% and 1% were added to a basal formulated diet (Table 73

1) for preparation of experimental diets. The ingredients were blended thoroughly in a mixer and 74

pelleted using a meat grinder equipped with a 2-mm die. The pelleted diets were air-dried and 75

stored in plastic bag at -4oC until further use. 76

Fish culture and feeding trial 77

Three hundred and sixty common carp larvae (550 ± 20 mg) were supplied from Dr. Keyvan 78

Marine Science and technology research center (Guilan province, Iran) adapted to experimental 79

conditions for one week. Thereafter, fish were randomly allocated into 9 tanks (500 L), 40 fish 80

per tank; 3 tanks per treatment. Water temperature, dissolved oxygen and pH maintained at 25.21 81

± 1.22 °C, 7.43 ± 0.13 mg L-1

and 7.43 ± 0.13, respectively. During the rearing trial (7 weeks), 82

the fish were hand-fed with experimental diets to apparent satiation three times a day at 7:30, 83

12:30 and 17:30 (Ebrahimi, Ouraji, Khalesi, Sudagar, Barari, Zarei Dangesaraki & Jani Khalili 84

2012). 85

Growth performance and survival 86

Growth performance parameters include; weight gain (%), specific growth rates (SGR), feed 87

conversion ratio (FCR), condition factor (CF) and survival rate were calculated according to the 88

following formulae: weight gain = W2 − W1, specific growth rate (SGR) = 100 [ln W2−ln W1]/T, 89

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condition factor (CF) = 100 × (body weight; g) / (body length; cm) 3

and feed conversion ratio 90

(FCR) = FO/WG. W1 is initial weight (g), W2 is final weight (g), T is time (days), FO is feed 91

offered (g) and WG is weight gain (g), survival = (Nf / N0)*100; where N0 is initial number of 92

fish and Nf is final number of fish. 93

Chemical analysis of diets and fish carcass composition 94

Triplicate samples of the experimental diets and 3 fish carcasses samples from each treatment 95

were obtained and chemical analysis performed according to AOAC (1990). Gross energy was 96

calculated using the conversion factors of 23.6, 39.5, and 17.0 kJ g-1

for protein, lipid, and 97

nitrogen-free extract (NFE), respectively as described elsewhere (Brett & Groves 1979). 98

Digestive enzyme activities assay 99

Three 24-h starved fish were sampled from each tank for digestive enzyme activities assay 100

(Yanbo & Zirong, 2006). The samples were prepared for assays according to the protocol 101

described elsewhere (Soleimani, Hoseinifar, Merrifield, Barati & Abadi, 2012). Protease activity 102

was determined based on the protocol suggested by Hidalgo, Urea & Sanz (1999) using casein 103

hydrolysis at pH 8. Amylase activity was quantified using starch as a substrate at 540 nm 104

(Bernfeld 1951) and lipase activity was determined by the measurement of fatty acids released 105

following enzymatic hydrolysis of triglycerides in a stabilized emulsion of olive oil (Yanbo & 106

Zirong 2006). All measured digestive enzyme activities in this experiment were expressed in 107

terms of U mg_1

protein min_1

. 108

Autochthonous intestinal microbiota 109

At the beginning and end of feeding trial 10 and 3 fish were sampled, respectively for 110

microbiological studies. Total viable counts of heterotrophic aerobic bacteria (TVC) and lactic 111

acid bacteria (LAB) levels were determined in intestinal samples of carp larvae. Sampling and 112

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preparation of intestine for investigation of the autochthonous intestinal microbiota was 113

performed as described previously (Hoseinifar, Mirvaghefi, Mojazi Amiri, Rostami & Merrifield 114

2011). Plate count agar (PCA) (Merck, Germany) and de Man, Rogosa and Sharpe (MRS) media 115

(Merck, Germany) were used for determination of total viable heterotrophic aerobic bacteria and 116

LAB, respectively. The plates were incubated at room temperature (25ºC) for 5 days and colony 117

forming units (CFU) g−1

were calculated from statistically viable plates (i.e. plates containing 118

30–300 colonies) (Rawling, Merrifield & Davies 2009). 119

Statistical analysis 120

Data analysis performed using one-way ANOVA followed by Duncan’s multiple range test, after 121

checking normality and homogeneity of means. Statistical analyses were conducted using SPSS 122

statistical package version 17.0 (SPSS Inc., Chicago, IL, USA). Mean values were considered 123

significantly different at P < 0.05. Data are presented as mean values ± SEM. 124

125

Result 126

The growth performance parameters of common carp larvae after 7 weeks feeding on 127

experimental diets are presented in Table 2. At the end of feeding trial there were no significant 128

differences between final weight, WG, SGR or FCR of larvae fed the control or 0.5 and 1% sc-129

FOS supplemented diets (P > 0.05). However, survival rate was significantly higher in larvae fed 130

1% dietary sc-FOS compared to 0.5% and control treatments (P < 0.05). 131

The results of carcass composition evaluation of larvae fed experimental diets contain low levels 132

of sc-FOS are shown in Table 3. Our results revealed the lowest body protein content and highest 133

lipid content in larvae fed 1% dietary sc-FOS (P < 0.05). No significant differences observed in 134

cases of body moisture and ash content between treatments (P > 0.05). 135

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The results of administration of low levels of dietary sc-FOS on digestive enzyme activities of 136

carp are presented in Figure 1. Lipase activity in common carp larvae fed 1% sc-FOS was 137

significantly (P < 0.05) higher compared to 0.5% sc-FOS and control treatment. The highest 138

amylase activity was observed in 1% sc-FOS group, which was significantly higher when 139

compared to other treatments (P < 0.05). However, feeding on different levels of sc-FOS had no 140

remarkable (P > 0.05) effects on the protease activity. 141

Figure 2 represents LAB and TVC levels of intestinal microbiota of carp larvae fed different 142

levels of dietary sc-FOS. No significant difference observed in TVC of prebiotic fed carp larvae 143

and control group (P > 0.05). However, LAB levels were significantly elevated after 7 weeks 144

feeding on sc-FOS (P < 0.05). There were no significant differences between 0.5 or 1% sc-FOS 145

treatments in case of LAB. 146

Discussion 147

It is now well-documented that administration of prebiotics which are functional dietary 148

supplements can improves growth performance, feed utilization, lipid metabolism, digestive 149

enzyme activities and stimulates immune response of fish and shellfish (Ringø et al., 2014). 150

However, such information is limited in larvae and larvae culture specially in Cyprinids 151

(Yousefian, Hedayatifard, Fahimi, Shikholeslami, Irani, Amirinia & Mousavi 2012; Genc, 152

Sengul & Genc 2013). The results of this study showed that administration of low levels (0.5 or 153

1%) of dietary sc-FOS had no remarkable effects on growth parameters and diet utilisation of 154

common carp larvae. Similarly, dietary FOS failed to improve growth performance of beluga 155

juveniles (Huso huso) (Hoseinifar et al., 2011) and Atlantic salmon (Salmo salar) (Grisdale-156

Helland, Helland & Gatlin III 2008). However, in contrast to our finding, administration of FOS 157

(or fructooligosaccharide) positively affects growth performance in blunt snout bream 158

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(Megalobrama amblycephala) fingerlings (Wu, Liu, Li, Xu, He, Li & Jiang 2013), stellate 159

sturgeon (Acipenser stellatus) juvenile (Akrami, Iri, Khoshbavar Rostami & Razeghi Mansour 160

2013), farmed rainbow trout (Oncorhynchus mykiss) (Ortiz, Rebolé, Velasco, Rodríguez, 161

Treviño, Tejedor & Alzueta 2013) and red drum (Sciaenops ocellatus) (Buentello, Neill & Gatlin 162

III 2010). The difference between results obtained in prebiotic studies can be attributed to 163

different methods of prebiotic administration, dosage levels, fermentability of the prebiotics and 164

differences in gut morphology and intestinal microbiota. 165

Dietary sc-FOS significantly affected body protein and lipid deposition in common carp larvae. 166

The lowest body protein deposition observed in experimental diet 1%. In accordance with the 167

results obtained in the present study, the lowest body protein deposition was observed in Atlantic 168

salmon (Salmo salar) fed with mannanoligosaccharides (Grisdale-Helland et al., 2008). 169

However, administration of FOS and mannanoligosaccharides caused increase of body protein 170

deposition in Japanese flounder (Paralichthys olivaceuce) (Ye, Wang, Li & Sun 2011) and 171

rainbow trout (Yilmaz, Genc & Genc 2007). The changes of body protein and lipid deposition of 172

fish can be due to changes in their synthesis, the rate of deposition in muscle, different growth 173

rates of fish at different ages and type of prebiotic as suggested previously (Abdel-Tawwab, 174

Abdel-Rahman & Ismael 2008) 175

The results of present study revealed that supplementation of low levels of dietary sc-FOS in 176

carp larvae diet significantly increased survival rate. The elevation of survival rate is of high 177

importance in culture of larvae and fry as production of larvae and fry appears to be a major 178

bottleneck of commercial aquaculture. In line with our results, similar results obtained following 179

administration of FOS in diets of cobia (Rachycentron canadum) larvae (Salze, McLean, 180

Schwarz & Craig 2008), rainbow trout (Staykov, Spring, Denev & Sweetman 2007) and beluga 181

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juvenile (Hoseinifar et al., 2011). The increase of survival rate of sc-FOS fed larvae is possibly 182

due to improved general health or immune status, as suggested previously (Soleimani et al., 183

2012). However, determination of the effects of dietary sc-FOS on immune system and health 184

status of carp larvae merit further research. 185

In this study, low levels of dietary sc-FOS had a significant effect on digestive enzyme activities 186

of carp larvae. Larvae fed 1% dietary sc-FOS had significantly higher amylase and lipease 187

compared to the control and 0.5% sc-FOS groups. The previous studies have shown that various 188

prebiotics had different effects on the activities of digestive enzyme. The positive effects of (1, 2 189

and 3%) dietary FOS and (50, 100 and 200 mg kg-1

) xylooligosaccharides have been reported on 190

Caspian roach (Rutilus rutilus) fry (Soleimani et al., 2012) and allogynogenetic crucian carp 191

(Carassius auratus gibelio) (Xu, Wang, Li & Lin 2009), respectively. However, dietary FOS 192

caused no significant changes on the activities of digestive enzymes (pepsin, trypsin, 193

chymotrypsin, aminopeptidase, a-amylase, lipase) in hybrid striped bass (Morone chrysops × M. 194

saxatilis) and red drum (Sciaenops ocellatus) (Anguiano, Pohlenz, Buentello & Gatlin 2013). 195

The increased digestive enzyme activities in the groups treated with prebiotic might be due to 196

elevated exogenous microbial activities (potentially modulated by the prebiotic). 197

In the present study, culture based analysis of the autochthonous intestinal microbiota of carp 198

larvae following administration of low levels of dietary sc-FOS revealed that although TVC were 199

not affected, LAB were significantly increased (Figure 2). Likewise, Akrami et al. (2013) and 200

Hoseinifar et al. (2011) stated that dietary FOS significantly increased LAB levels in intestinal 201

microbiota of stellate sturgeon (Acipenser stellatus) and beluga (Huso huso) juvenile, 202

respectively. LABs have been considered as beneficial components of the fish intestinal 203

microbiota (Ringø & Gatesoupe 1998). However, they are minor constitute of intestinal 204

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microbiota of several fish species and administration of prebiotics have been suggested for 205

elevation of LABs in intestinal microbiota (Merrifield et al., 2014). Our results showed that 206

dietary sc-FOS can modulate intestinal microbiota of carp larvae toward potentially beneficial 207

communities (i.e LABs); although determinations of affected strain using molecular techniques 208

(e.g DGGE) requires further research. 209

In conclusion, the present study revealed that growth performance, survival, digestive enzyme 210

activities as well as intestinal microbiota of carp larvae can be modulated through administration 211

of low levels of dietary sc-FOS. However, still further research should be undertaken on different 212

aspect of prebiotics administration in carp fry and larvae culture. 213

Acknowledgments 214

This work was supported by Young Researchers Club, Lahijan Branch. The author wish to thank 215

Orafti company managers (Raffinerie Tirlemontoise, Tienen, Belgium) for their support and 216

provision of the prebiotic and Prof. Van Loo, Dr. Mahious for their kind help. 217

References: 218

Abdel-Tawwab M., AM Abdel-Rahman & Ismael N.E. (2008) Evaluation of commercial live bakers’ yeast, 219

Saccharomyces cerevisiae as a growth and immunity promoter for Fry Nile tilapia, Oreochromis niloticus 220

(L.) challenged in situ with Aeromonas hydrophila. Aquaculture, 280, 185-189. 221

Akrami R., Iri, Y., Khoshbavar Rostami, H & Razeghi Mansour, M. (2013) Effect of dietary supplementation of 222

fructooligosaccharide (FOS) on growth performance, survival, lactobacillus bacterial population and 223

hemato-immunological parameters of stellate sturgeon (Acipenser stellatus) juvenile. Fish & Shellfish 224

Immunology, 35, 1235-1239. 225

Anguiano M., Pohlenz C., Buentello A & Gatlin D.M. (2013) The effects of prebiotics on the digestive enzymes and 226

gut histomorphology of red drum (Sciaenops ocellatus) and hybrid striped bass (Morone chrysops× M. 227

saxatilis). British Journal of Nutrition, 109, 623-629. 228

AOAC (1990) Official methods of analysis, Association of Analytical Chemists, , Arlington, VA. 229

Page 10 of 20



For Review O

nly

11

Bernfeld P. ( 1951) Amylases α and β. In: Methods in Enzymology (eds Colowick P, Kaplan NO). Academic Press., 230

New York, NY, pp. 149–157. 231

Brett J & Groves T. (1979) Physiological Energetics. Fish physiology, 8, 279-352. 232

Buentello JA., Neill W.H & Gatlin III D.M. (2010) Effects of dietary prebiotics on the growth, feed efficiency and 233

non specific immunity of juvenile red drum Sciaenops ocellatus fed soybean based diets. Aquaculture 234

Research, 41, 411-418. 235

Cabello F.C. (2004) Antibiotics and aquaculture in Chile: Implications for human and animal health. Revista Medica 236

De Chile, 132, 1001-1006. 237

Denev S., Staykov Y., Moutafchieva R & Beev G. (2009) Microbial ecology of the gastrointestinal tract of fish and 238

the potential application of probiotics and prebiotics in finfish aquaculture. International Aquatic Research, 239

1, 1-29. 240

Dimitroglou A., Merrifield D.L., Carnevali O., Picchietti S., Avella M., Daniels C., Guroy D & Davies S.J. (2011) 241

Microbial manipulations to improve fish health and production-a Mediterranean perspective. Fish & 242

Shellfish Immunology, 30, 1-16. 243

Ebrahimi G., Ouraji H., Khalesi M., Sudagar M., Barari A., Zarei Dangesaraki M & Jani Khalili K. (2012) Effects of 244

a prebiotic, Immunogen®, on feed utilization, body composition, immunity and resistance to Aeromonas 245

hydrophila infection in the common carp Cyprinus carpio (Linnaeus) fingerlings. Journal of Animal 246

Physiology and Animal Nutrition, 96, 591-599. 247

Gatesoupe F.J. (2008) Updating the importance of lactic acid bacteria in fish farming: Natural occurrence and 248

probiotic treatments. Journal of Molecular Microbiology and Biotechnology, 14, 107-114. 249

Genc M.A., Sengul H & Genc E. (2013) Effects of dietary mannan oligosaccaharides on growth, body composition, 250

intestine and liver histology of the common carp (Cyprinus carpio L.) Fry. In: Aquaculture Europe 2013. 251

WAS, Trondheim, Norway. 252

Grisdale-Helland B., Helland S.J & Gatlin III D.M. (2008) The effects of dietary supplementation with 253

mannanoligosaccharide, fructooligosaccharide or galactooligosaccharide on the growth and feed utilization 254

of Atlantic salmon (Salmo salar). Aquaculture, 283, 163-167. 255

Hidalgo M., Urea E & Sanz A. (1999) Comparative study of digestive enzymes in fish with different nutritional 256

habits. Proteolytic and amylase activities. Aquaculture, 170, 267-283. 257

Page 11 of 20



For Review O

nly

12

Hoseinifar S.H., Mirvaghefi A., Mojazi Amiri B., Rostami H.K & Merrifield D.L. (2011) The effects of 258

oligofructose on growth performance, survival and autochthonous intestinal microbiota of beluga (Huso 259

huso) juveniles. Aquaculture Nutrition, 17, 498-504. 260

Merrifield D., Balcázar J., Daniels C., Zhou Z., Carnevali O., Sun Y., Hoseinifar S & Ringø E (2014) Indigenous 261

lactic acid bacteria in fish and crustaceans. In: Aquaculture Nutrition: Gut Health, Probiotics and 262

Prebiotics (ed. by Ringø E, Merrifield DL). Wiley-Blackwell scientific Publication, London. 263

Merrifield D.L., Dimitroglou A., Foey A., Davies S.J., Baker R.T.M., Bøgwald J., Castex M & Ringø E. (2010) The 264

current status and future focus of probiotic and prebiotic applications for salmonids. Aquaculture, 302, 1-265

18. 266

Niness K.R (1999) Inulin and oligofructose: What are they? Journal of Nutrition, 129, 1402S-1406S. 267

Ortiz L., Rebolé A., Velasco S., Rodríguez M., Treviño J., Tejedor J & Alzueta C. (2013) Effects of inulin and 268

fructooligosaccharides on growth performance, body chemical composition and intestinal microbiota of 269

farmed rainbow trout (Oncorhynchus mykiss). Aquaculture Nutrition, 19, 475–482. 270

Rawling M.D., Merrifield DL & Davies S.J. (2009) Preliminary assessment of dietary supplementation of Sangrovit 271

on red tilapia (Oreochromis niloticus) growth performance and health. Aquaculture, 294, 118-122. 272

Ringø E., Dimitroglou A., Hoseinifar S.H & Davies S.J. (2014) Prebiotics in finfish: an update. In: Aquaculture 273

Nutrition: Gut Health, Probiotics and Prebiotics (ed. by Ringø E, Merrifield DL). Wiley-Blackwell 274

scientific Publication, London. 275

Ringø E & Gatesoupe F.J. (1998) Lactic acid bacteria in fish: a review. Aquaculture, 160, 177-203. 276

Ringø E., Olsen R.E., Gifstad T.Ø., Dalmo R.A., Amlund H., Hemre G.I & Bakke A.M. (2010) Prebiotics in 277

aquaculture: a review. Aquaculture Nutrition, 16, 117-136. 278

Salze G., McLean E., Schwarz M.H & Craig S.R. (2008) Dietary mannan oligosaccharide enhances salinity 279

tolerance and gut development of larval cobia. Aquaculture, 274, 148-152. 280

Soleimani N., Hoseinifar S.H., Merrifield D.L., Barati M & Abadi Z.H. (2012) Dietary supplementation of 281

fructooligosaccharide (FOS) improves the innate immune response, stress resistance, digestive enzyme 282

activities and growth performance of Caspian roach (Rutilus rutilus) fry. Fish & Shellfish Immunology, 32, 283

316-321. 284

Page 12 of 20



For Review O

nly

13

Staykov Y., Spring P., Denev S & Sweetman J. (2007) Effect of a mannan oligosaccharide on the growth 285

performance and immune status of rainbow trout (Oncorhynchus mykiss). Aquaculture International, 15, 286

153-161. 287

Wu Y., Liu W.B., Li H.Y., Xu W.N., He J.X., Li X.F & Jiang G.Z. (2013) Effects of dietary supplementation of 288

fructooligosaccharide on growth performance, body composition, intestinal enzymes activities and 289

histology of blunt snout bream (Megalobrama amblycephala) fingerlings. Aquaculture Nutrition, 19, 886-290

894. 291

Xu B., Wang Y., Li J & Lin Q. (2009) Effect of prebiotic xylooligosaccharides on growth performances and 292

digestive enzyme activities of allogynogenetic crucian carp (Carassius auratus gibelio). Fish physiology 293

and biochemistry, 35, 351-357. 294

Yanbo W & Zirong X. (2006) Effect of probiotics for common carp (Cyprinus carpio) based on growth performance 295

and digestive enzyme activities. Animal Feed Science and Technology, 127, 283-292. 296

Ye J.D., Wang K., Li F.D & Sun Y.Z. (2011) Single or combined effects of fructo‐and mannan oligosaccharide 297

supplements and Bacillus clausii on the growth, feed utilization, body composition, digestive enzyme 298

activity, innate immune response and lipid metabolism of the Japanese flounder Paralichthys olivaceus. 299

Aquaculture Nutrition, 17, e902-e911. 300

Yilmaz E., Genc M.A & Genc E. (2007) Effects of dietary mannan oligosaccharides on growth, body composition, 301

and intestine and liver histology of rainbow trout, Oncorhynchus mykiss. Israeli Journal of Aquaculture-302

Bamidgeh, 59, 182-188. 303

Yousefian M., Hedayatifard M., Fahimi S., Shikholeslami M., Irani M., Amirinia C & Mousavi S.E. (2012) Effect of 304

Prebiotic supplementation on growth performance and serum biochemical parameters of Kutum (Rutilus 305

frisii kutum) Fries. Asian Journal of Animal & Veterinary Advances, 7, 1-9. 306

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308

309

310

311

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Table 1. Dietary formulations and proximate composition of basal diets (%).

Ingredient Control

Fish meal 40.0

Wheat flour 21.0

Soybean meal 13.5

Gluten 5.5

Soybean oil 6.0

Fish oil 6.0

Mineral premix1 3.0

Vitamin premix1 2.0

Binder2 2.0

Anti fungi3 0.5

Antioxidant4 0.5

Proximate analysis

Dry matter 89.60

Crude protein 38.23

Crude lipid 10.22

Ash 3.40

Gross Energy (MJ kg-1) 17.58

1 Premix detailed by (Eshaghzadeh et al., 2014)

2 Amet binder ™, Mehr Taban-e- Yazd, Iran

3 ToxiBan antifungal (Vet-A-Mix, Shenan- doah, IA)

4 Butylated hydroxytoluene (BHT) (Merck, Germany)

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Table 2. Growth performances of common carp larvae fed low levels of dietary sc-FOS; control diet

(0% sc-FOS), 0.5% sc-FOS and 1% sc-FOS for 7 weeks. Data represent means from 3 replicates per

treatment. Values are presented as the mean ± SE

Control sc-FOS 0.5% sc-FOS 1%

Final weight (g) 2.22 ± 0.1 2.37 ± 0.09 2.31 ± 0.1

Weight gain (g) 1.63 ± 0.07 1.77 ± 0.1 1.74 ± 0.06

SGR 2.72 ± 0.08 2.82 ± 0.2 2.84 ± 0.04

CF 2.25 ± 0.08 2.02 ± 0.07 2.33 ± 0.1

FCR 1.22 ± 0.08 1.12 ± 0.07 1.15 ± 0.1

Survival (%) 71.0 ± 0.2a

75.0 ± 0.3a

89.3 ± 0.1b

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Table 3. Carcass proximate composition of common carp larvae fed low levels of dietary sc-FOS;

control diet (0% sc-FOS), 0.5% sc-FOS and 1% sc-FOS for 7 weeks. Data represent means from 3

replicates per treatment. Values in a row with the different superscripts denote significant difference.

Values are presented as the mean ± SE.

Control Oligo 0.5% Oligo 1%

Moisture 79.80 ± 0.09 79.59 ± 0.05 79.62 ± 0.06

Crude protein 14.296 ± 0.33b

12.84 ± 0.1a

12.51± 0.1a

Crude lipid 6.844 ± 0.10a

7.95 ± 0.31b

8.24± 0.12b

Ash 4.625 ± 1.12 4.03 ± 0.37 3.96 ± 0.44

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Figure 1. Digestive enzymes activities [protease (a), amylase (b) and lipase (c)] of common carp

larvae fed a basal diet (control) and two diets supplemented with 0.5% or 1% sc-FOS for 7

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weeks. Data represent the mean ± SEM. Bars assigned with different superscripts are

significantly different (P < 0.05).

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Figure 2. Total culturable autochthonous bacterial (A) and autochthonous lactic acid bacteria

(LAB) (B) levels (log CFU g-1 intestine) of carp larvae fed low levels of dietary sc-FOS; control

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diet (0% sc-FOS), 0.5% sc-FOS and 1% sc-FOS for 7 weeks. Bars assigned with different

superscripts are significantly different (P < 0.05); Values are presented as the mean ± SE.

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Modulation of growth performances, survival, digestive enzyme activities and intestinal microbiota...

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