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5

Calcium carbonate

5

5

5

5

5

Moisture

8.1

7.6

7.8

8.2

7.7

Crude protein

47.2

46.9

46.8

47.1

47

Crude lipid

14.9

14.9

15.1

15

15.1

Crude starch

11.9

12

11.8

11.9

12.1

Gross energy (kcal/g)4

4.55

4.54

4.54

4.56

4.57

Digestible energy (kcal/g)5

3.98

3.96

3.97

3.98

3.99

permix: (mg or IU/kg of diet) Vitamin A (as acetate) 1600000 IU; vitamin D3, 400000 IU; choline chloride.12000; niacin, 4000; riboflavin, 8000; pyridoxine, 4000; folic acid, 2000; vitamin B12, 8000; biotin, 1; inositol, 20000; vitamin C, 60000; vitamin H2, 2.4; vitamin B2, 8000; vitamin K3, 2000; vitamin E,40000. 3Mineral mixture (g/kg): zinc, 12.5 g; iron, 26 g;

manganese, 15.8 g; copper, 4.2 g; cobalt, 0.48 g; selenium, 2 g; iodine, 1 g. 4Calculated on the basis of 5.64, 9.43, and 4.11 (kcal/g diet) for protein, fat, and carbohydrate, respectively (NRC 1993). 5Calculated using apparent coefficients of digestibility of 0.9, 0.85, and 0.8 for crude protein, crude fat, and carbohydrates (NFE), respectively.

As depicted in table 2, no significant differences were detected in growth indices of fish fed FO and those were fed on diet containing CO and CSLO. However, total replacement of fish oil with SO and LO

resulted in decreased WG, SGR and DGR. Fish fed FO, CO and CSLO

showed similar FCR, whereas FCR was significantly higher in SO and LO treatments. There was also a good accordance between FCR and WG among different treatments; a higher WG was recorded in a group with lower FCR. HSI was significantly lower in CO group, while it was significantly higher in CSLO group in comparison to control. CO and LO had significantly lower VSI than others (P< 0.05). CF was significantly higher in fish fed diet containing 100% vegetable oil (CO, SO. LO and CSLO), comparing to FO (P<0.05, table 2). As it is clear from table 2, no significant differences were detected in protein PPV among treatments.

PER decreased significantly in SO and LO groups compared to those fed fish oil. However, PER from CO and CSLO groups did not show any significant differences compared FO (P< 0.05). Replacement of fish oil with vegetable oils (CO, SO, LO and CSLO groups) resulted in significantly lower LPV compared to those fed fish oil (P< 0.05). LER in CO, SO and LO was significantly lower compared FO group (P< 0.05).

Proximate compositions of fillets were shown in table 3. Replacement of fish oil with vegetable oils (CO, SO, LO and CSLO groups) resulted in significantly higher muscle protein and ash content. However, plant oil inclusion led to lower fillet lipid content in comparison to FO (P< 0.05).

Table 2 Growth and feed utilization indices of different experimental groups

Dietary treatments1

Performance parameters FO CO SO LO CSLO

Final body weight (g)

69.9±2.3a

69.6±0.4a

58.2±0.5b

61.5±2.5b

68.2±2.2a

Weight Gain (g/fish)

54.8±2.2a

54.4±0.2a

43.0±0.4b

46.1±2.3b

53.1±2.1a

Feed intake (g/fish)

55.4±7.8ab

49.9±0.3a

38.8±3.8ab

39.4±3.9bcd

54.0±6.0abc

FCR2

1.01±0.03b

0.98±0.03b

1.2±0.05a

1.2±0.01a

1.02±0.04b

DGR3

2.88±0.07a

2.87±0.01a

2.45±0.01b

2.55±0.07b

2.83±0.08a

SGR4

1.17±0.02a

1.16±0.01a

1.02±0.01b

1.05±0.02b

1.14±0.03a

HIS5

1.51±0.02bc

1.47±0.04c

1.57±0.07b

1.50±0.03bc

1.75±0.06a

VSI6

15.2±1.0a

11.2±0.2c

15.9±0.3a

13.7±0.9b

16.1±0.6a

CF7

1.08±0.03b

1.22±0.02a

1.20±0.03a

1.17±0.02a

1.20±0.09a

PPV8

0.39±0.04ab

0.37±0.2ab

0.35±0.02b

0.37±0.02ab

0.41±0.03a

PER9

2.19±0.21a

2.01±0.67ab

1.84±0.07b

1.90±0.13b

2.20±0.15a

LPV10

0.12±0.01a

0.08±0.01bc

0.07±0.01cd

0.06±0.01d

0.10±0.01b

LER11

5.04±0.50a

4.43±0.04b

4.11±0.29b

4.26±0.21b

5.07±0.35a

Values are means ±S.D. Values with the same superscripts within the same row are not significantly different (P < 0.05). 1See Table 1 for diet abbreviations.2FCR, food conversion ratio; 3DGR, Daily growth rate; 4SGR, specific growth rate; 5HIS, Hepatosomatic index; 6VSI, viscerosomatic index; 7CF, Condition factor. 8PPV, protein production value; 9PER, protein efficiency ratio; 10LPV, lipid production value; 11LER, lipid efficiency ratio.

Table 3 Proximate composition of muscle (%) ofdifferentexperimental groups

Dietary treatments

Component -

FO        CO        SO        LO CSLO

Moisture (%)

77.3±2.0ab

70.7±2.0b

77.4±3.1a

78.9±2.4a

76.0±4.5a

Protein (%DM)

74.5±0.7c

75.1±0.6b

76.9±0.2ab

77.9±0.8a

76.9±0.8ab

Lipid (%DM)

11.3±0.3a

9.5±0.3bc

9.4±0.3bc

8.7±0.9c

9.7±0.4b

Ash (%DM)

7.3±0.2b

7.9±0.2a

7.8±0.9a

7.8±0.3a

7.7±0.1a

Values are means ± S.D. Values not sharing the same superscript within the same row were significantly different (P < 0.05).

Discussions

The present study showed that it is possible to totally replace fish oil by CO and CSLO without any apparent undesirable effects on fish growth performance and feed utilization indices. While it was stated that LO is a promising candidate for salmonids (Bendiksen and Jobling, 2004), this study did not support the previous results. This contrasting results might be due to the fact that within each plant species, many variants are available that differ in their fatty acid profiles and consequently their nutritional value. Canola oil, another popular oilseed, has yielded promising results. Higher PPV and PER of CO and CSLO implied a better dietary protein utilization due to dietary lipid utilization as a main energy source to sustain growth without increasing lipid deposition in fish in comparison to FO (Tocher, 2003). These finding were in good accordance with lower SFA and higher MUFA content of VO compared to FO (data not shown). However, the probable rationale behind the increased protein utilization remained to be clarified yet. According to the table 3, it seemed that plant oil sources led to lower fillet lipid contents due to the higher amounts of monoenes (MUFA) and lower saturated fatty acid (SFA) contents present in VO in comparison to FO (R0sj0 et al., 2000; Nordrum et al., 2003; Regost et al., 2003). However, Ferreira et al. (2011) reported contrasting results in Nile tilapia fed non-fish meal based diets with various plant oil sources due to the fact that MUFA and SFA content of VO diets were completely different comparing to the diets used in the present. It was stated balancing LA, ALA, EPA, and DHA in CSLO could promote growth similar to that seen with FO so that mixed plant oils are more promising for fish oil replacement efforts. Apart from fish health and performance, substitution of marine oils with VO has been shown to be effective in reducing the levels of dioxins, dioxin-like polychlorinated biphenyls and organo-chlorinated pesticides in fish products (Bell et al., 2005). In conclusion, it seems that CO and CSLO are promising sources of dietary lipid for rainbow trout; however, thorough conclusion needs precise and complete examinations. This study is still ongoing and further biochemical and histological assessments are being undertaken.

Bell, J.G., McGhee, F., Dick, J.R., Tocher, D.R., 2005. Dioxin anddioxin-like polychlorinated biphenyls (PCBs) in Scottishfarmed salmon (Salmosalar): effects of replacement of dietarymarine fish oil with vegetable oils. Aquaculture 243:305-314.

Bell, J.G., McEvoy, J., Tocher, D.R., McGhee, F., Campbell, P.J.,Sargent, J.R., 2001.Replacement of fish oil with rape seedoil in diets of Atlantic salmon (Salmosalar) affects tissuelipid compositions and hepatocyte fatty acid metabolism.Journal of Nutrition 131: 1535-1543.

Bell, J.G., Tocher, D.R:, Henderson, R.J., Dick, J.R., Crampton, V.O., 2003b. Altered fatty acid compositions in Atlantic salmon (Salmosalar) fed diets containing linseed and rapeseed oils can be partially restored by a subsequent fish oil finishing diet. Journal of Nutrition 133: 2793-2801.

Bendiksen, E., Jobling, M., 2004.Effects of temperature and feed composition on essential fatty acid (n-3 and n-6) retention in Atlantic salmon (Salmosalar L.) parr. Fish Physiology and Biochemistry 29: 133-140.

Drew M.D., Ogunkoya A.E., Janz D.M. & Van Kessel A.G. (2007) Dietary influence of replacing fish meal and oil with canola protein concentrate and vegetable oils on growth performance, fatty acid composition and organochlorine residues in rainbow trout (Oncorhynchusmykiss). Aquaculture 267, 260-268.

Miller M.R., Nichols P.D. & Carter C.G. (2007) Replacement of dietary fish oil for

Atlantic salmon parr (Salmo salar L.) with a stearidonic acid containing oil has no effect on omega-3 long-chain polyunsaturated fatty acid concentrations. Comp Biochem Physiol B Biochem Mol Biol 146: 197-206.

Nordrum, S., Olli, J.J., R0sj0, C., Holm, H., Krogdahl, A., 2003. Effects of graded levels of medium chain triglycerides and cysteine on growth, digestive processes and nutrient utilization in sea water reared Atlantic salmon (Salmosalar, L.) under ad libitum feeding regime. Aquaculture Nutrition 9: 263-274.

NRC (1993) Nutrient Requirements of Fish.National Academy Press, Washington, DC.

Regost, C., Arzel, J., Cardinal, M., Rosenlund, G., Kaushik, S.J., 2003.Total

replacement of fish oil by soybean or linseed oil with a return to fish oil in Turbot (Psetta maxima). 2. Flesh quality properties. Aquaculture 220: 737-747.

R0sj0, C., Nordrum, S., Olli, J.J., Krogdahl, A.,Ruyter, B., Holm, H., 2000. Lipid

digestibility and metabolism in Atlantic salmon (Salmosalar) fed medium-chain triglycerides. Aquaculture 190: 65-76.

Stubhang, I., 0. Lie and B.E. Torstensen. (2007). Fatty acid productive value and □­oxidation capacity in Atlantic salmon tissues (Salmo salar L.) fed on different lipid sources along the whole growth period. Aquaculture Nutrition 13: 145 -

155.

Tacon, A. G.J., Hsan, M.R., Allan, G., El-Sayed, A.-F.M., Jackson, A., Kaushik, S.J.,

Ng, W-K., Suresh, V., Viana, M.T., 2012. Aquaculture feeds: addressing the long-term sustainability of the sector. In FAO/NACA.Farming the Waters for People and Food. R.P. Subasinghe, J.R. Arthur, D.M. Bartley, S.S. De Silva, M. Halwart, N. Hishamunda, C.V. Mohan & P. Sorgeloos, (Eds.) Proceedings of the Global Conference on Aquaculture 2010, Phuket, Thailand. 22-25 September 2010. FAO, Rome and NACA, Bangkok. 896 pp.

Tocher, D.R., Bell, J.G., McGhee, F., Dick, J., Fonseca-Madrigal, J., 2003.Effects of

dietary lipid level and vegetable oil on fatty acid metabolism in Atlantic salmon (Salmosalar L.) over the whole production cycle. Fish Physiology and Biochemistry 29: 193-209.

Effect of Tribulus terrestris extract on sex reversal of Fighting fish (Betta splendens) by Tribulus terrestris extract

Janalizadeh Elham1, Manouchehri Hamed2' Reza Changizi3, M.Hosseinifard3.

1 MSc. graduated from Islamic Azad University of Babol Fisheries and member of

Young Researchers Club 2,3,4 Department of Fisheries, Islamic Azad University of Babol, Iran.

Abstract

The effects of Tribulus terrestris extract on sex reversal of fighter fish(Betta splendens) evaluated through this survey via enrichment method by TT extract. The aim of this study was application of a photogenic composition on the B.splendens. The male fighter is more valuable and attractive with nice body in contrast with female. Tribulus terrestris is a natural and non-toxic herb that cause increasing testosterone levels in humans and animals. The plant extracts were prepared and different doses of TT (0.0, 0.01, 0.05 , 0.1 g/l ) were examined for sex changing of B.splendens. According to results, in the enrichment method, fighters larvae have been examined for 4 months by TT extract in diet (Biomar pellet enrichment with the extract of Tribulus trrestris). Among The treatment doses, the treatment with 0.05 g/l showed a significant effect with a high percentage of males (%72.71) (p<0.05). Survival percentage in treatment groups and control groups were equally at a high rate of %81.84. Herbal extract of TT had a positive impact on survival B.splendens. All fish was found in the TT than in the control group showed increasing growth and more performance. TT experimental groups with concentrations of 0.05 g/l B.splendens showed a high rate of growth significantly (p <0.05).

Keywords: plant extract, Tribulus trrestris, masculinizatin, Betta splendens

Tribulus Terrestris (Order Zygophyllacea) is an herb that is widely distributed in China, Japan, Korea, the western part of Asia, the southern part of Europe and Africa. It has been shown to raise testosterone levels safety and naturally and is rumoured to be the secret behind the success of many top Bulgarian weightlifters (Bucci,2000). Using TT for human and animals improved libido and spermatogenesis (Tomova et al., 1981). It has been used to increase testosterone levels and improve athletic performance (Bocci, 2000; Adaikan et al., 2000; Adimoelja, 2000; Gauthaman, 2002). Testosterone is a sex hormone and its role in sexuality is unequivocal. Embryonic differentiation of the larvae into a male and its subsequent growth along this line is essentially due to the presence of physiological amounts of testosterone in the body. Testosterone is the biosynthetic precursor to both androgens and estrogens in teleosts (Baroiller et al., 1999; Devlin and Nagahama, 2002). However, in addition to testosterone, 11-oxygenated androgens are commonly found at high concentrations in the plasma and tissues of male fish and are generally considered to be the most important physiologycal androgens in males ( Borg, 1994 ). TT contains a number of different substances known as steroidal saponins. The saponin in TT thought to be responsible for its effect on testosterone levels is known as protodioscin (Adimoelja and Adaikan, 1997; Ganzera et al., 2001; Joshi et al., 2005).

The live-bearing fighter, Betta spelendens, is a popular aquarium fish. Aquarists use them for obtaining new morphas with variable colours. The male of this species are more attractive than the females therefore maintenance and breeding of male populations has generated a great amount of commercial interest. Masculinization of B. splendens can be produced by direct synthetic hormonal treatment that is efficient and straightforward ( Takahashi, 1975; Kavumpuratha and Pandian, 1992, 1993 ). However synthetic hormones are more expensive than plant extracts and their fate in water and sediment is providing information on the potential risks of using synthetic hormones ( Contreras-Sanchez et al., 2001 ). An alternate technique for commercial production of all male

production is perhaps to use plant extracts. Therefore the objective of the present study is to investigate the effect of TT on sex reversal in B. splendens.

Materials and Methods

Extraction

Ethanol extraction method was performed using the Rotary (Vamanu Emanuel et al., 2011).

Preparation for productive replication Feeding treatments

In this experiment, fish were fed by green water for 2 months the they were fed by Artemia and enriched pellet extracts of Tribulus terrestris until they reached maturity (4 months). Doses of Tribulus terrestris extract for enriched pellet were 0.01, 0.05 and 0.1 g/l. Tribulus terrestris extract was mix and spray on dried pellet. Parameters such as motility, growth, body length, deformity, survival, number of significant differences between male and have been evaluated in this research. Our criterion for measuring the mobility experiments were conducted to determine the mobility and balance of the natural swimming fish.

Statistical analysis

At the end of the experiment after collecting the entire Kolmograph-smirnov test of normality of the data was determined. Data were analyzed by SPSS software. Treatments to Tables and graphs were plotted using Excel 2007.

Results

The method of feeding Tribulus terrestris extract treated with pellets enriched with the highest percent survival and the highest percentage of males in the 0.05 g/l has been viewed( Fig1-1). Compared with males the percentage of survival was lower. High mortality was observed in 0.1g/l significantly.

Control 0.01 0.05 0.1

Treatment

In Fig 1 .percent of maculinization and survival of fish feed enriched with extracts of TT Percent survival rate in the treated male 0.1 g/l compared with the control treatments and treatments with a dose of 0.05 g/l was decreased. Percent of survival in the control group, 0.01, 0.05 and 0.1 g/l were %86.9, % 75.47, % 77.82 and %13.33 respectively.Percentage of male were %50.86, % 52.08, % 72.71 and %44.44 respectively.

Discussion

Many of the active ingredients in drugs are used can be extracted from plants. Nowaday the use of plants as medicine is not something that should be devoted to a few countries, But many countries, including the developed countries of the plants, formulated drugs are marketed and pure active ingredients and pharmaceutical companies to produce the drug available to the plants studied And thereby a significant share of its exports to this account (Torabi Goodarzi, 1999).

The immersion method was used by Santhakumar and Thavamani, 2002. They used 17a-methyl testosterone hormone on fish Btta splendens

fighter resulted to %98 survival of and %71 of masculinization . The way immersion guppy fish Poecilia reticulate using Tribulus terrestris extract % 80 achieved masculinization (Sehriban et al., 2007). The study found that all treatments reduced survival with increasing dose. In parallel with it Derakhshan et al (2007) effects of intraperitoneal administration of therapeutic doses of the herb on rat kidneys were examined, than The severity of the lesions observed in mice that had been treated for 35 days Far more acute and severe than the group that received 28 days of treatment with Tribulus terrestris extract.

The growth rate in this study was higher than control, but at the end of maturity ( 4 months) compared with control subjects did not differ, but research on musculunization of Cichlasoma nigrofasciatum fish is done, increase the success rate in all experimental groups with Tribulus terrestris extract to compared with the control group (Sehriban, 2007). In this study, the highest dose (0.1 g/l ) treatments have been seen to have some deformity and decreased survival rates and the best dose, 0.05 g/l. Similar to the results with increasing doses of 17 a-methyl testosterone in Betta splendens survival decreased to %76 at a dose of 900 mg/l is reached, but the treatments of 1000 mg/l deformities suffered and died within 2-3 days (Santhakumar and Thavamani, 2002). The research was conducted, it has been observed in all dose treatments of sex. Although the Betta splendens hormonal immersion method at the lowest dose of 100 mg/l were not able to change sexual (Santhakumar and Thavamani, 2002).

Conclusion

Therefore, this study has shown that Tribulus terrestris extract at a dose of 0.05 g/l increase in the percentage of male and percentage of survival declined with increasing dose. Therefore, the dose of 0.1 g/l compared with group of control survival decreases, but the percentage of male is higher. According to the survey results, using industrial food enrich with Tribulus terrestris to masculinization other aquarium fish and farmed to because than the hormonal methods is safer and less costly suggested

Adaikan,P.G., k. Gauthaman, R.N.V.prasad and S.C.Ng, 2000. Proerectile pharmacological effect of Tribulus terrestris on the rabbit corpus cavernosum.Ann. Acad. Med. Singapore, 29:22-26.

Adimoelja, A.and P.G. Adaikan, 1997. Protodioscin from herbal plant Tribulus terrestris L. improves male sexual functions possibily via DHEA. Int. J . Impot.

Res., 9:64.

Ahmadi far. E., (2007). Effects of feeding enriched Artemia urmiana Argus growth, percent survival and resistance to thermal stress in Persian sturgeon larvae Acipencer persicus, Iranian Journal of Biology, p 509.

Baroiller, J.F., Y . Guigen and A. Fostier, 1999 . Endocrine and environmental aspects of sex differentiation in fish. Cell. Life Sci., 55:910-931 .

Bucci, L.R., 2000. Selrcted herbals and human exercise Performanc. Am . J . Clin.

Nutr., 72:624-636.

Contreras-Sanchez, W.M. M.s. Fitzpatrick and C.B Schteck.2001. Fate of Methyl Testosterone in the pond Environment: Detection of Mtin Pond Soil from a CRSPsite. In:Enghteenth Dnnaual Technical Report. Gupta, A., K. McElwee, D. Burright,X Cummings and H. Egna (Eds.), Oregon State University,Corvallis, Organ, PP: 79-82.

Derakhshan. A., (2007), Empirical studies of histological lesions induced by intraperitoneal injection Tribulus terrestris extract plant, Fifteenth Congress of Veterinary Medicine.

Gauthaman K, Adaikan PG, Prasad RNV . Goh VHH, Ng SC (2000) Changes in

hormonal parameters secondary to intravenous administration of Tribulus terrestris extract in primates. Int J Import Res 12:6

Santhakumar-K; Thavamani-J : Journal of experimental zoology.: 2002 , 606-616

Sehriban-C; Funda-T : Biological Sciences for Scientific Information.: 2007 , 718-725

Torabi goodarzi. M., (1999), and Natural Resources Research Center of Qom, Journal of Aquaculture No. 7, pp. 34-33.

Vamanu. E., Tropical Journal of Pharmaceutical Research December 2011; 10 (6): 777-783

Increased resistance of rainbow trout (Oncorhynchus mykiss) fed with Mannan (as a prebiotic) against Yersinia ruckeri

Rezvaneh Jenabi1*, Amir Tukmechi2, Saeed Meshkini1, Maria Mohseni1

1 Department of Fishery, Faculty of Natural Resources, Urmia University

2 Department of Pathobiology, Artemia and Aquatic Animals Research Institute, Urmia University

* Corresponding authors: rezvanehjenabi @ Yahoo.com Abstract

The aim of this study was to evaluate the efficacy of prebiotic Mannan oligosaccharides (0, 2.5 and 5 g/kg of diet) against bacterial infection in rainbow trout. For this purpose, fish were fed with diet containing mentioned concentrations as a prebiotic for 45 days. Then, fish were exposed intraperitoneally with pathogenic strains of Yersinia ruckeri and the results were compared with a control group. The results showed that feeding with Manan significantly (P<0.05) improved rainbow trout resistant against Yersinia ruckeri.

Keywords: peribiotic Manan oligosaccharide, Rainbow trout and Yersinia ruckeri.

Introduction

Prebiotics are nondigestible carbohydrates (NDCs) that selectively stimulate the growth and metabolism of health-promoting bacteria already present in the host gut (Ahmadifar et al., 2004). Improving and protecting fish health in commercial production practices is a major factor in the aquaculture industry. The uses of dietary supplements that are involved in growth and enhance the immune system are strategies that can be helpful in improving health, resistance to the stress and pathogens. Prebiotic beneficially affects the host by selectively stimulating the growth and/or activity of one or a limited number of

bacteria in the colon, which can improve the host health (Gibson and Roberfroid, 1995). Criteria which allow the classification of a food ingredient as a prebiotic include: (1) it must be neither hydrolyzed nor absorbed in the upper part of the gastrointestinal tract, (2) selective fermentation by one or a limited number of potentially beneficial bacteria in the colon, and (3) alteration of the composition of the colonic microbiota toward a healthier composition (Fooks and Gibson, 2002). Among the established prebiotics such as fructooligosaccharide, transgalactooligosaccharide, inulin and mannan oligosaccharide (MOS), MOS is most commonly used as the dietary supplementation for fish and crustacean species (Sang etal., 2009). MOS is a glucomannoprotein complex derived from the cell wall of yeast (Saccharomyces cerevisiae) (Sang and Fotedar, 2010). Some studies were conducted to investigate the effects of MOS on the growth and immune response of fish and crustaceans. It can be pointed the positive effects of inulin and Oligofructose prebiotic in diets of Siberian sturgeon (Acipenser baerii) and African catfish (Acipenser baerii) on Immune parameters (Mahious et al., 2005). Also, research showed Gro Biotic-A® (a commercial prebiotic) has led to rainbow trout resistant to a variety of bacterial infections. Although specified beneficial effects of prebiotics, very little research has been done in this field (Mahhious and Ollevier, 2005). The aim of this study was to investigate the resistance of rainbow trout fed with MOS against experimental infection by Yersinia ruckeri.

Materials and Methods

Fish and experimental design

For hundared rainbow trout (10± 1.2 g mean weight) were prepared from a local fish farm, Urmia, Iran. Immediately fish were transferred to the laboratory with oxygen supply and adapted to the laboratory conditions for 10 days. Then, they were divided into three groups included: control, fed with commercial pellet (Faradaneh, Iranl), group 2 (2.5 grams of MOS per kilogram of diet) and group 3 (5 grams of MOS per kilogram of diet). Fish culture was conducted in 300 liter PVC tanks and forty fish located in each tank. Trial was conducted for 45 days and

water quality parameters such as dissolved oxygen (10.3 mg/l), temperature (14.1±2 C), and pH value (7.5 ± 0.4) were measured every day during the experimental period. Desired amount of MOS (2.5 and 5 gr) dissolved in 10 ml of sterile normal saline and sprayed with 5 ml sunflower oil on diet. Then, allowed to diet dried at the room temperature and at a clean place. Control group only received commercial pellet diet that sprayed with normal saline and sunflower oil.

Experimental Bacterial challenge

After 45 days 10 fish per each tank randomly were exposed to pathogenic strain of Yersinia ruckeri. Each fish injected intraperitoneally with 5x107 CFU/ml of bacteria and they were located at 90 liters tanks. Within 7 days fish were examined daily and their mortality was recorded daily.

Statistical analysis

The results were subjected to analysis of variance (ANOVA) followed by Tukey test to compare different treatments using the SPSS 15. Correlation coefficients were significant with P< 0.05. Determination of the LD50 was conducted using probit analysis program.

Results and Discussion

After 7 days, the results showed:

Day

Mortality caused by the Yersinia ruckeri

Fish were dark-bodied and hyperemia was seen around the mouth and fins. Mortality in the control group began the first day but in Second and third groups began the third day The control group had a higher Mortality and the third group had a fewer Mortality than other groups. The results showed that treatments with MOS feeding significantly (p <0.05) was more resistant than the control group. In the third group because of the high level of MOS, fish were compared to a control group and the second group has more resistant and less mortality. The results of the current study indicate that dietary supplementation with 2.5 and 5 g/kg of Mannan oligosaccharide had significant effects on Oncorhynchus mykiss increase resistance because the prebiotic Mannan oligosaccharide enhances the immune system.

These results are consistent with the results of Torrecillas (2007) that showed 0, 2 and 4 g of MOS per kg diet in European sea bass (Dicentrachus labrax) increase in the amount of Lysozyme resistance against Vibrio alginolyticus bacterial infection.

Staykov used 2 g.kg-1 MOS to evaluate the effect on growth performance and immune status of rainbow trout (Oncorhynchus mykiss) and found significantly reduced mortality, and improved indicators of immune status compared with controls (Staykov et al., 2007).

In contrast to positive effects of MOS, studies by Sang (2009) on Cherax tenuimanus showed feeding with 0.2 and 0.4 g/kg mannan oligosaccharide could significantly enhance fish resistance against environmental stresses and bacterial infections. This study showed that the use of Mannan oligosaccharide in rainbow trout (Oncorhynchus mykiss) could increase fish resistance against Yersinia ruckeri.

Ahmadifar, E., Azari Takami, G., Sudagar, M., (2004). Growth performance, survival and immunostimulation, of Beluga (Huso huso) juvenile following dietary administration of alginic acid (Ergosan). Pak J Nut 8(3):227-232.

Fooks, L.J., Gibson, G.R., (2002). Probiotic as a modulators of the gut flora. Br J Nutr

1:39-49.

Gibson, G.R., Roberfroid, M.B., (1995). Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr 125:1401-1412.

Mahious AS, Ollevier F (2005) Probiotics and prebiotics in aquaculture: review. In: 1st Regional workshop on techniques for enrichment of live food for use in larviculture AAARC, pp 17-26, Urmia.

Sang, H., ky, L., Fotedar, R., (2009). Dietary supplementation of mannan oligosaccharide improves the immune responses and survival of marron, Cherax tenuimanus (Smith, 1912) when challenged with different stressors. Fish and Shellfish Immunol 27: 341-348.

Sang, H.M., Fotedar, R., (2010). Effects of mannan oligosaccharide dietary supplementation on performances of the tropical spiny lobster juvenile (Panulirus ornatus). Fish and Shellfish Immunol 22: 1-7.

Staykov, Y., Spring, P,. Denev, S et al (2007). Effect of a mannan oligosaccharide on the growth performance and immune status of rainbow trout (Oncorhynchus mykiss). Aquacult Inter 15:153-161.

Torrecillas, S., Makol, A., Caballero, D., Robaina, L., Real, F., Sweetman, J., Tort, L., and Izqueirdo, M.S., (2007). Immune stimulution and improved infection resistance in european sea bass (Dicentrarchus labrax) fed mannan oligosaccharides. Fish and Shellfish Immunol 23: 969-981.

Effect of dietary mannan oligosaccharide (MOS) on growth performance, survival in rainbow trout (Oncorhynchus mykiss)

Rezvaneh Jenabi1*, Amir Tukmechi2

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