EFFECTS OF MANNAN-OLIGOSACCHARIDE ON GROWTH, SURVIVAL AND DISEASE RESISTANCE OF NILE TILAPIA (OREOCHROMIS NILOTICUS LINNAEUS) FRY

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1 8 th Interntionl Symposium on Tilpi in Aquculture EFFECTS OF MANNAN-OLIGOSACCHARIDE ON GROWTH, SURVIVAL AND DISEASE RESISTANCE OF NILE TILAPIA (OREOCHROMIS NILOTICUS LINNAEUS) FRY C. SAMRONGPAN*, N. AREECHON, R. YOONPUNDH AND P. SRISAPOOME Dept of Aquculture, Fculty of Fisheries, Ksetsrt University,Bngkok 10900, Thilnd Abstrct Effects of mnnn-oligoscchride (MOS) on growth, survivl nd disese resistnce of Nile tilpi (Oreochromis niloticus Linneus) fry were studied. Four concentrtions of MOS t 0, 2, 4 nd 6 g/kg feed were given to tilpi fry for 21 dys. There were significnt differences mongst verge weight, length nd verge dily growth (ADG) of fry (P<0.05) with the vlues of 0.78±0.16, 0.79±0.19, 0.85±0.20 nd 0.90±0.19 g nd 2.86±0.24, 2.88±0.20, 2.96±0.25 nd 3.03±0.22 cm nd 0.037±7, 0.037±9, 0.040±0.010 nd 0.042±9 g/dy respectively. Weight, length nd ADG of tilpi fed with 4 nd 6 g/kg MOS were significntly higher thn the control (P<0 05).. There were no significnt differences mongst feed conversion rtio (FCR) nd survivl rte of ll experimentl nd control group (P>0.05). Tilpi tht were rised with MOS t ll concentrtions did show significntly higher resistnce ginst pthogenic bcteri, Streptococcus glctie (P<0.05) with mortlity of 43.33±20.82, 3.33±5.77, ± nd ± % for control, 2, 4 nd 6 g/kg MOS respectively. These results indicted the benefit of MOS s feed supplement for tilpi fry in term of growth nd disese resistnce. Further study on the effects of MOS nd its ppliction in lrge tilpi will be conducted. Keywords: Nile Tilpi, mnnn-oligoscchride, growth, survivl, disese resistnce *CorrespondentE-mil: chubby_cupid@hotmil.com INTRODUCTION Nile tilpi (Oreochromis niloticus Linneus) hs been the most common freshwter fish being cultured ll round the world. In Thilnd, this species count for more thn 35% of the whole production from freshwter quculture. There re two min systems of tilpi culture in this country including pond nd cge culture which require millions of fry ech yer. Cge culture cn supply the better qulity of the hrvest especilly for export. Fry production is the vitl fctor for tilpi culture. Mny frmers prefer to use ll-mle tilpi fry becuse they cn grow fster nd eliminte the over-popultion problem. Frmers lso cn predict the production nd hrvesting

2 346 EFFECTS OF MANNAN-OLIGOSACCHARIDE ON GROWTH, SURVIVAL AND DISEASE RESISTANCE OF NILE TILAPIA (OREOCHROMIS NILOTICUS LINNAEUS) FRY period. Besides the price issue, tilpi frmers lso encounter with disese nd growth problem. The most common disese of tilpi is cused by Grm positive bcteri nmely, Streptococcus glctie (Driny et l., 2007). Streptococcosis is very common during summer seson in conjunction with limited nd poor qulity wter supply. Infected fish showed sluggish behvior, spirl swimming nd protruded eyes (exophthlmi). These round-shped grm positive bcteri cn be isolted from internl orgns nd from the brin. Frmers re using licensed ntibiotics to cope with the outbreks but the results re still not stisfctory. Growth is nother issue of interested for tilpi intensive cultures both in htchery nd grow-out stge. In htchery, during sex-reversl period of 21 dys, growth, size distribution nd helth ppers to be the trgets of reserch. In this report, mnnn-oligoscchride (MOS) ws pplied during the htchery stge to determine the following prmeters: -growth nd survivl rte -disese resistnce ginst S. glctie MATERIALS AND METHODS Experimentl Designs Tilpi lrv with just-disppered yolk sc with verge weight of g were used for this experiment. The experiment ws conducted in 100-L glss quri holding 80 liters of dechlorinted tp wter. Ech qurium ws stocked with 100 tilpi lrv. Continuous ertion ws pplied throughout the tril. Powder feed (40% protein) ws used s bsl diet. This experiment ws conducted by Completely Rndomized Design with 4 experimentl groups with three replictes s follows: Control Exp # 1 Exp # 2 Exp # 3 Bsl diet + 60 ppm 17 α methyl testosterone (17 α-mt) Bsl diet + 60 ppm 17 α-mt + 2 g/kg MOS Bsl diet + 60 ppm 17 α-mt + 4 g/kg MOS Bsl diet + 60 ppm 17 α-mt + 6 g/kg MOS This experiment ws designed ccording to the procedure of sex-reversl htchery stge of Nile Tilpi in which 60 ppm of 17 α-mt will be pplied for 21 dys. Fish were fed 5 times dy between 8.00 m nd 5.00 pm. Wter were chnged t 70% rte t every two dys.

3 C. SAMRONGPAN et l. 347 Determined Prmeters After 21 dys of feeding tril, the following prmeters were determined nd sttisticlly compred by Anlysis of Vrince nd Duncn s New Multiple Rnge Test t p = 0.05 (Boonom, 2007). -Growth. Thirty percents of ech replicte were mesured for weight nd length. Averge dily growth (ADG) nd feed conversion rtio (FCR) were lso compred. -Survivl rte -Disese resistnce ginst pthogenic Streptococcus glctie. Ten fish from ech replicte were chllenged by immersion in 10 8 cfu/ml bcteril suspension for 6 hrs nd trnsferred to norml wter. Rte of mortlity were observed nd compred for 7 dys. Bcteril isoltion ws performed from ded or moribund fish to confirm the cuse of infection. RESULTS Growth nd Survivl After 21 dys of feeding tril, tilpi fry fed with 4 nd 6 g/kg MOS showed significntly better weight, length nd ADG thn control nd 2 g/kg (P<0.05) (Tble 1; Fig 1, 2 nd 3). However, FCR were not significntly different (P>0.05) (Tble 1; Fig 4) Tble 1. Effect of different concentrtions of mnnn-oligoscchride on weight, stndrd length, verge dily growth nd feed conversion rtio of Nile tilpi fry fter n experimentl period of 21 dys (Men ± S.D.) Tretment Weight (g) Stndrd Length (cm) Averge dily growth (g/dy) Feed conversion rtio Control 0.78± ± ±7 1.42± g/kg 0.79± ± ±9 1.39± g/kg 0.85±0.20 b 2.96±0.25 b 0.040±0.010 b 1.28± g/kg 0.90±0.19 b 3.03±0.22 b 0.042±9 b 1.12±0.09 Note: Significnt difference ws indicted by different superscript letter within the sme column t P=0.05

4 348 EFFECTS OF MANNAN-OLIGOSACCHARIDE ON GROWTH, SURVIVAL AND DISEASE RESISTANCE OF NILE TILAPIA (OREOCHROMIS NILOTICUS LINNAEUS) FRY Weight (g) b b Control 2 g/kg 4 g/kg 6 g/kg Tretment Figure 1. Effect of different concentrtions of mnnn-oligoscchride on weight (Men ± S.D.) of Nile tilpi fry Stndrd length (cm) b b Control 2 g/kg 4 g/kg 6 g/kg Tretment Figure 2. Effect of different concentrtions of mnnn-oligoscchride on stndrd length (Men ± S.D.) of Nile tilpi fry

5 C. SAMRONGPAN et l. 349 Averge dily growth (g/dy) b b Control 2 g/kg 4 g/kg 6 g/kg Tretment Figure 3. Effect of different concentrtions of mnnn-oligoscchride on verge dily growth (Men ± S.D.) of Nile tilpi fry Feed conversion rte Control 2 g/kg 4 g/kg 6 g/kg Tretment Figure 4. Effect of different concentrtions of mnnn-oligoscchride on feed conversion rte (Men ± S.D.) of Nile tilpi fry Survivl rtes were not significntly different (P>0.05) fter 21 dy tril rnging from % (Tble 2 nd Fig 5)

6 350 EFFECTS OF MANNAN-OLIGOSACCHARIDE ON GROWTH, SURVIVAL AND DISEASE RESISTANCE OF NILE TILAPIA (OREOCHROMIS NILOTICUS LINNAEUS) FRY Tble 2. Effect of different concentrtions of mnnn-oligoscchride on survivl rte (%) of Nile tilpi fry fter n experimentl period of 21 dys (Men ± S.D.) Tretment Survivl rte (%) Control 89.67± g/kg 94.00± g/kg 95.67± g/kg 96.33±6.35 Note: No significnt difference ws detected (P>0.05) Survivl rte (%) Control 2 g/kg 4 g/kg 6 g/kg Tretment Figure 5. Effect of different concentrtions of mnnn-oligoscchride on survivl rte (%) (Men ± S.D.) of Nile tilpi fry Resistnce ginst Streptococcus glctie It ws clerly shown from this experiment tht MOS could enhnce the immunity of Nile Tilpi fry. All experimentl groups fed with MOS hd significntly lowered mortlity rte thn the control one fter chllenged with virulent S. glctie (P<0.05). Tilpi tht were fed with 4 nd 6 g/kg MOS did not hve ny mortlity while there ws 3.33% mortlity in 2 g/kg MOS group nd 43.33% of control fish died (Tble 3 nd Fig 6) Tble 3. Effect of different concentrtions of mnnn-oligoscchride on disese resistnce ginst Streptococcus glctie (Men ± S.D.) of Nile tilpi fry Tretment Men percent mortlity (± S.D.) Control 43.33± g/kg 3.33±5.77 b 4 g/kg ± b 6 g/kg ± b Note: Significnt difference ws indicted by different superscript letter within the sme column t P=0.05

7 C. SAMRONGPAN et l. 351 Mortlity rte (%) b b b Control 2 g/kg 4 g/kg 6 g/kg Tretment Figure 6. Effect of different concentrtions of mnnn-oligoscchride on disese resistnce ginst St reptococcus glctie (Men ± S.D.) of Nile tilpi fry DISCUSSION AND CONCLUSION This study clerly indicted the benefit of mnnn-oligoscchride for tilpi fry. With 4 nd 6 g/kg MOS in bsl diet during 21 dy of sex reversl process, tilpi hd significntly better growth rte in term of weight, length nd verge dily growth. The study on MOS with other freshwter species such s Europen ctfish (Silurus glnis) juveniles (Bogut et l., 2006) hve shown similr improvement in growth from 22 g to 76 g in the control group nd 83 g in the MOS group, 9.7% higher verge body weight (P<0.01). The mortlity decresed from % to 16.67% (P<0 01).. Juvenile crp rered in tnks t the University of Osijek, Croti lso showed similr improvement in weight gin (Culjk et l., 2006). The diet used in these trils ws 39.91% crude protein nd 4.51% lipid nd in the MOS treted groups with n inclusion rte of 0.6%. The fish grew from n verge weight of 5.28 g to g in the control nd g in the MOS treted group, 24% higher weight gin (P<0.01). The FCR significntly improved in the MOS treted groups, decresing from 2.06 to 1.60 (P<0.05) nd the mortlity lso decresed from 50.0% to 16.7% (P<0.01). In rinbow trout trils (Stykov et l., 2005 & b) with 0.2% MOS inclusion rte in commercil feed resulted in significntly incresed verge weight of 13.7% (P<1) in fish grown from 30 g to just under 100 g. Mortlities nd FCR were significntly lower in the MOS treted groups. The FCR decresed from 0.91 in the control group to 0.83 in the MOS treted groups (P<0 05).. Mortlities decresed from 1.68% in the control groups to 0.58% in the MOS treted groups (P< 1). At the University of Ls Plms of Grn Cnri in Spin, 35 g se bss (Dicentrrchus

8 352 EFFECTS OF MANNAN-OLIGOSACCHARIDE ON GROWTH, SURVIVAL AND DISEASE RESISTANCE OF NILE TILAPIA (OREOCHROMIS NILOTICUS LINNAEUS) FRY lbrx) juveniles showed improved specific growth rte (SGR) nd reltive growth which incresed significntly by bout 10% (P<0.05) fter fed with MOS t 0.2 nd 0.4% inclusion rtes (Torrecills nd Izquierdo, 2006). In the MOS treted se bss, higher feed intke ws observed when compred to the control, however t the end of the experimentl period ll the groups hd similr FCR. In this present study, survivl rte nd feed conversion rtio of control nd ll MOS-treted Nile tilpi (O. niloticus) fry were not significntly different fter the 21- dy tril. This might be relted to the length of the tril. However, ll concentrtions of MOS could significntly enhnce the resistnce ginst pthogenic Streptococcus glctie. In common crp (Cyprinus crpio L.) nd rinbow trout (Oncorhynchus mykiss) tht were fed with MOS, hd increse bctericidl ctivity, ntibody titres, lysozyme concentrtion (Lie, 1985), lterntive pthwy of complement ctivity (APCA) (Sotirov, 1986) nd clssicl pthwy of complement ctivity (CPCA) (Stelzr nd Stein, 1971) when compred with the control. Similr s 0.2% nd 0.4% of MOS in Europen se bss (D. lbrx) tril (Torrecills et l., 2007) which could ctivte immune system when chllenge test with pthogenic Vibrio lginolyticus. Further experiments must be conducted to clrify the ction mechnisms of MOS with immune system, s well s the optimum feeding period nd dose for low cpitl production. The results from this study clerly showed the benefits of mnnnoligoscchride (MOS) s feed supplement on growth nd disese resistnce ginst pthogenic bcteri, Streptococcus glctie during the 21-dy nursing period of Nile tilpi fry. On-going reserch is being conducted to investigte the benefits of MOS on immune functions of Nile tilpi. ACKNOWLEDGMENTS Specil thnks to Kmphengsn Freshwter Fisheries Sttion, Fculty of Fisheries, Ksetsrt University, Nkhonpthom, Thilnd for supplying the fish nd to Alltech Biotechnology Corportion, Bngkok, Thilnd for supplying MOS. REFERENCES 1. Bogut, I., Z. Milkovic, J. Pvlicevic nd D. Petrovic Effect of Bio-Mos on performnce nd helth of Europen ctfish, Sillirus glnis. Proceedings of Alltech s 22 nd Annul Symposium April 23-26, Lexington, KY, USA. 2. Boonom, C Experimentl Design. Dept of Sttistic, Fculty of Science, Ksetsrt University, Bngkok, Thilnd.

9 C. SAMRONGPAN et l Culjk, V., I. Bogut, E. Hs-Schon, Z. Milkovic nd K. Cnecki Effect of Bio-Mos on performnce nd helth of juvenile crp. Proceedings of Alltech s 22 nd Annul Symposium April 23-26, Lexington, KY, USA. 4. Driny, J., N. Areechon nd P. Srispoome Study on identifiction nd ntimicrobil sensitivity test of Streptococus spp. isolted from Nile tilpi (Oreochromis niloticus). Proceeding of 44 th Ksetsrt University Annul Conference 30 Jn 2 Feb, Bngkok, Thilnd. 5. Lie, O Improved gr plte ssys of bovine lysozyme nd hemolytic complement ctivity, pp In Mrkers of Resistnce to Infection in Diry Cttle. Disserttion, Oslo, Norwy, Ntionl Veterinry Institute. 6. Sotirov, L. K Method for determintion of the lterntive pthwy of complement ctivtion in some nimls nd mn, pp In Fourth Scientific Conference of Agriculture. StrZgor. 7. Stykov, Y., S. Denev nd P. Spring The effects of mnnn oligoscchride (Bio-Mos) on the growth rte nd immune function of rinbow trout (Slmo girdneri irideus G.) growth in net cges, pp In B. Howell nd R. Flos (eds.). Lessons from the pst to optimise the future. Europen Aquculture Society, Specil Publiction No.35. June Stykov, Y., S. Denev nd P. Spring. 2005b.The effects of mnnn oligoscchride (Bio-Mos) on the growth rte nd immune function of rinbow trout (Slmo girdneri irideus G.) growth in net cges, pp In B. Howell nd R. Flos (eds.). Lessons from the pst to optimise the future. Europen Aquculture Society, Specil Publiction No.35. June Stelzr, A. nd G. Stein Moglichkikten zur hmolytishchen Aktivittsmessung von Gesmtkomplement Theoretische Grundlngen und methodische Hinweise. Wissenschftliche Zeitschrift der Friedrich Schiler Universitt Uen. Mthemtisch Nturwissenschftlische Reihe 20, Jg H6: Torrecills, S. nd M. S. Izquierdo The effect of Bio-Mos on Europen se bss (Dicentrrchus lbrx) juveniles: growth performnce nd feed utiliztion. Proceedings of Alltech s 22 nd Annul Symposium April 23-26, Lexington, KY, USA. 11. Torrecills S., A. Mkol, M. Jcbllero, D. Montero, L. Robin, F. Rel, J. Sweetmn, L. Tort nd M. S. Izquierdo Immune stimultion nd improved infection resistnce in Europen se bss (Dicentrrchus lbrx) fed mnnn oligoscchrides. Fish & Shellfish Immunol. 23: