한수지 48(4), 466-473, 215 Originl Article Koren J Fish Aqut Sci 48(4),466-473,215 홍해삼 (Stichopus jponicus) 치삼의성장및면역증강을위한면역증강사료첨가제의개발 남궁진 안경진 1 여인규 * 제주대학교해양생명과학과, 1 ( 주 ) 준원지비아이 Growth nd Physiologicl Effects of Immunity Feed Additives on the Juvenile Red Se Cucumer Stichopus jponicus Jin Nmgung, Kyoung-Jin Ahn 1 nd In-Kyu Yeo* Deprtment of Mrine Life Science, Jeju Ntionl University, Jeju 64234, Kore 1 Junwon GBI Co. Ltd., JeJu 63169, Kore The juvenile red vrint of the se cucumer Stichopus jponicus is distriuted worldwide. It is vlule food source in Kore, Chin, nd Jpn. Mjor issues in frming the juvenile red vrint se cucumer include growth rtes nd disese resistnce. In recent yers, studies hve focused on feed dditives to enhnce immune system nd helth. In this study, we used common diet (CD), nutritionl diet (ND; mixed nutritionl supplements), nd n immunity diet (ID; mixed proiotics, spirulin, nd levn) nd compred the growth rtes nd immunity of juvenile red vrint se cucumers fed the experimentl diets for 12 weeks. The growth nd survivl rtes in the ID group were significntly incresed (P>.5). This suggests tht the feed dditives positively influenced immunity nd growth in the ID group. However, the immune ctivity ws exhiit stilizing effect, nd further investigtion of immune effects is required. Key words: Red Se cucumer, Immunostimulnt, Proiotics, Spirulin, Levn 서론 (Stichopus jponicus) (Choo, 28).,., (Kn-no nd Kijim, 23; Choo, 28)..,, (Yng et l., 25)., (cellulose) (Xi et l., 212; Shi et l., 213). (Ersmus et l., 1997; Ten Doeschte nd Coyne, 28).,, (Choi, 214).,. (Xito et l., 214). http://dx.doi.org/1.5657/kfas.215.466 Koren J Fish Aqut Sci 48(4) 466-473, August 215 This is n Open Access rticle distriuted under the terms of the Cretive Commons Attriution Non-Commercil Licens (http://cretivecommons.org/licenses/y-nc/3./) which permits unrestricted non-commercil use, distriution, nd reproduction in ny medium, provided the originl work is properly cited. Received 3 July 215; Revised 8 August 215; Accepted 11 August 215 *Corresponding uthor: Tel: +82. 64.754. 3474 Fx: +82. 64. 756. 3493 E-mil ddress: ikyeo99@jejunu.c.kr Copyright 215 The Koren Society of Fisheries nd Aqutic Science 466 pissn:374-8111, eissn:2287-8815
홍해삼치삼의면역증강제섭이에따른영향 467. (Zho et l., 21; M et l., 213; Wng et l., 214) (proiotics) (FAO/WHO, 21). (De schrijver nd Ollevier, 2; Ten Doeschte nd Coyne, 28; Yu et l., 29; Zokeifr et l., 212), (Sugit et l., 1992; Vine et l., 26). (Preiotics). (pro) (pre) (syniotics) (Cross ML, 22; Morelli et l., 23). ph (Gison nd Roerfroid, 1995; El-Ngr et l., 22; Akin et l., 27; Adeol et l., 214), (Kng et l., 29, Sriknth et l., 215).,,.,,,,. 실험동물및실험환경 재료및방법 214 1 27 215 1 27 12, 3. (S. jponicus), 182 2.4.1 g. 1 2.4.3 m 3, 1 lux., 4 (YSI, YSI Incorported, Ohio, USA),, (Dissolved oxygen, D.O) ph (Tle 1). 사료제작및급이 (common diet, CD) CD (snyiho/ynti, duoli feed co., Ltd.) 1 kg 2% (nutrition diet, ND) 1 kg 15%, (Bcillus polyfermenticus) 3 1 9 CFU.1% (immunity diet, ID). CD Tle 2. 12 1 3 g. 성장도분석, 4. 12. Specific growth rte (%)=(log e finl wt-log e initil wt)/dys, Weight gin (%)=(finl weight-initil weight) 1 /initil weight. 홍해삼내다당류분석 Tle 1.Wter environment of experiment tnk for 12 weeks Week Wter temp ( ) Slinity (psu) Dissolved oxygen (DO; mg/l) ph 21.±.35 3.61±3.32 6.13±.13 7.64±.7 4 15.18±.6 33.47±.3 8.35±.3 7.88±.15 8 13.67±.47 34.45±.1 8.37±.14 7.85±.25 12 11.6±.75 31.8±5.77 9.14±.33 7.82±.11 Tle 2. Proximte composition (%) of common diet (CD) for red se cucumer Stichopus jponicus Moisture (%) 5.33 Crude protein (%) 21.7 Crude ft (%) 1.32 Crude sh (%) 34.65
468 남궁진ㆍ안경진ㆍ여인규. -7 3 72. 4. 2 M TFA (trifluorocetic cid) (.2 µm) HPAEC-PAD system (Dionex, USA). 18 mm NOH, CroPc TM PA1. 면역활성분석 1. 1 ml 1.5 ml tue 7 g 15,. -7. Lowry et l. (1952), Bovin serum lumin (BSA, sigm, USA) stndrd 75 nm. Lysozyme 15 L 96 well plte, 15 L Micrococcus lysodeikticus solution (.2 mg/ml,.1m PBS, ph=6.8) 25 5 54 nm, 5 lysis. Penoloxydse (PO) Ashid nd Dohke (198). 96 well plte 15 L phospte uffer (ph 6.) 15 L, 3,4-dihydroxy-L-phenyllnine (L- DOPA, sigm, USA) 15 L 25 25 49 nm 5..1. 통계처리 4 12. CD 73.1.84%, ND 7.9.32%, ID 76.9.5% ID (Fig. 1A, P<.5). 2.4.1 g, CD 4.6.5 g, ND 5.3.62 g ID 6.1.53 g (Fig. 1B, P<.5). 2.4.1 g CD 12 91.67 17.2%, ND 118.75 35.2% ID 155.6 31.7% (Tle 3, P<.5). CD.21.3%, ND.27.8%, ID.34.5% ID,, (A) (B) Survil rte (%) Body weight (g) 1 9 8 7 7 6 5 4 CD ND ID CD ND ID c c c SPSS version 12 (SPSS Inc., USA) One-wy ANOVA- test Duncn s multiple test P<.5. 3 2 2 4 6 8 1 12 14 Week 생존율및성장률측정 결과및고찰 Fig. 1. Survivl rte (A) nd ody weight (B) of the juvenile red type se cucumer Stichopus jponicus experimentl diet fed fter 12 weeks. Experimentl groups: common diet (CD), nutrition diet (ND) nd immunity diet (ID). Dt re expressed s men±se, different superscript letters (,, c) indicte significnt difference etween the vlues (P<.5).
홍해삼치삼의면역증강제섭이에따른영향 469 (P<.5, Tle 3).,,, (strglus memrnceus polyscchrides), (yest polyscchride), ( -glucns) (vitmin) (Sun et l., 28; Gu et l., 21; Zho et l., 21; M et l., 213; Wng et l., 214; Sriknth et l., 215)., (Xi et l., 212; Shi et l., 213; Choi, 214). Hliotis mide. (Ersmus et l., 1997; Ten Doeschte nd Coyne, 28; A. Newj-Fyzul, 214)., (Gison nd Roerfroid, 1995;Kontul et l., 1999; Srel et l., 23; Gu et l., 211; Boonnuntnsrn et l., 215). ID,.. Tle 3.Growth performnce of red se cucumer Stichopus jponicus fter 12 weeks CD ND ID Initil weight (g/fish) 1 2.4±.1 2.4±.17 2.4±.13 Finl weight (g/fish) 1 4.6±.5 5.3±.62 6.1±.53 c Weight gin (%) 2 91.6±17.2 118.8±35.2 156.±31.7 c Specific growth rte (%) 3.21±.3.27±.8.34±.5 c 1 Stndrd weight = totl weight (g)/ fish. 2 Specific growth rte (%) = (log e finl wt-log e initil wt)/dys. 3 Weight gin (%) = (finl weight-initil weight)*1 /initil weight. CD=common diet. ND= nutrition diet. ID= immunity diet. Men within ech line followed y the different superscript letters (,, c) indicte significnt difference etween the vlues (P<.5). 홍해삼내다당류분석 (Mourão et l., 1996; Yu et l., 215) fructose glucose. Fructose CD 8.84 1.35 mg/1 ml, ND 8.6 2.7 mg/1 ml ID 9.25 3.14 mg/1 ml (Fig. 2A). Glucose CD 9.2 2. mg/1 ml, ND 8.21.95 mg/1 ml ID 1.4 1.85 mg/1 ml (Fig. 2B). fructose glucose (P>.5), ID. fructose glucose fructose glucose, (A) (B) Concentrtion of fructose (mg/1 mg) Concentrtion of fructose (mg/1 mg) 14 12 1 8 6 4 2 14 12 1 8 6 4 2 CD Fig. 2. Glucose (A) nd fructose (B) concentrtion in ody well of the juvenile red type se cucumer Stichopus jponicus. Experimentl groups: common diet (CD), nutrition diet (ND) nd immunity diet (ID). Dt re expressed s men±se, different superscript letters (,, c) indicte significnt difference etween the vlues (P<.5). ND ID
47 남궁진ㆍ안경진ㆍ여인규 (P>.5). fructose glucose (Rirkhwd et l., 27; Rehm, 29), ID.. 면역활성분석, lysozyme penoloxydse (PO). CD 56. 1 mg/ ml, ND 66.1 24 mg/ml ID 66.5 19 mg/ml (Fig. 3), CD ND ID (P>.5). (coelomocytes), (Glinski nd Jrosz., 2; Rmírez-Gómez et l.,21)., C3, AMP (Antimicroil Peptides), Toll like receptors, lectins, PO, lysozyme (Jing et l., 214; Xue et l., 215). cytokine IL-1 IL-6 Protein level in coelomic fluid (mg/ml) 1 8 6 4 2 Fig. 3. Protein level in coelomic fluid of the juvenile red type se cucumer Stichopus jponicus. Experimentl groups: common diet (CD), nutrition diet (ND) nd immunity diet (ID). Dt re expressed s men±se, different superscript letters (,, c) indicte significnt difference etween the vlues (P<.5). CD ND ID, (protein fmily) (Gross PS et l.,1999; Rmírez-Gómez nd Grcí-Arrrás.,21).,... (P>.5) ND ID., lysozyme CD 29.54 9.2 U/mg, ND 49.52 7.2 U/mg, ID 14.9 3.2 U/mg ID (Fig. 4A, P<.5). Lysozyme, (Surh nd Shoo, 28; Nyk, 21). lysozyme (Wng et l., 28), lysozyme (Rirkhwd et l., 27; Dhech et l., 211; Xue et l., 215). ID 12 lysozyme. lysozyme. M et l. (213) lysozyme,., Tin et l. (215) c-type lysozyme. ID lysozyme ID, ID lysozyme. lysozyme
홍해삼치삼의면역증강제섭이에따른영향 471. PO, CD 14.7 6.5 U/mg, ND 17.2 8. U/mg ID 16.2 3.8 U/mg CD ND ID (Fig. 4B, P>.5). PO lysozyme (Jing et l., 214). Mgndóttir et l. (26) PO nti-protese 1 -ntiprotese, 2 -ntiprotese 2 -mcrogloloulin PO (Mgndóttir et l., 26). PO (A) (B) Lysozyme ctivity (U/mg) PO ctivity (U/mg) 6 5 4 3 2 3 25 2 15 1 5 CD Fig. 4. Lysozyme ctivity (A) nd penoloxydse ctivity (B) in coelomic fluid of the juvenile red type se cucumer Stichopus jponicus. Experimentl groups: CD; common diet, ND; nutrition diet nd ID; immunity diet. Dt re expressed s men±se, different superscript letters (,, c) indicte, significnt difference etween the vlues (P<.5). ND c ID.,,,.. 사사 214 (No.C289). References Adeol OO, Corcorn O nd Morgn WA. 214. Syniotics: The impct of potentil preiotics inulin, lctulose nd lctoionic cid on the survivl nd growth of lctocilli proiotics. J Funct Foods 1, 75-84. http://dx.doi.org/1.116/j. jff.214.5.1. Akin MB, Akin MS nd Kirmci Z. 27. Effects of inulin nd sugr levels on the viility of yogurt nd proiotic cteri nd the physicl nd sensory chrcteristics in proiotic ice crem. Food Chem 14, 93-99. http://dx.doi.org/1.116/j. foodchem.26.11.3. Boonnuntnsrn S, Wongssk U, Pitksong T nd Chijmrus S. 215. Effects of dietry supplementtion with β-glucn nd syniotics on growth, hemolymph chemistry, nd intestinl microiot nd morphology in the Pcific white shrimp. Aquc NutArticle first pulished online. http://dx.doi.org/1.1111/nu.1232. Choi GS. 214. Effects of Experimentl formulted diets mixture rtio on growth nd ody composition of se cucumer Stichopus jponicus. Mster Theses, Inh University, Incheon, Kore. Choo P S. 28. Popultion sttus, fisheries nd trde of se cucumers in Asi. Se Cucumers. A Glol Rev Fish Trde 516, 81-118. Cross ML. 22. Microes versus microes: immune signls generted y proiotic lctocilli nd their role in protection ginst microil pthogens. FEMS Immunol Med Microiol 34, 245-253. http://dx.doi.org/1.1111/j.1574-695x.22.t632.x. Dhech I, Belghith KS, Hmden K, Feki A, Belghith H nd Mejdou H. 211. Orl dministrtion of levn polyscchride reduces the lloxn-induced oxidtive stress in rts. Int J Biol Mcromol 49, 942-947. http://dx.doi.org/1.116/j.
472 남궁진ㆍ안경진ㆍ여인규 ijiomc.211.8.11. De Schrijver R nd Ollevier F. 2. Protein digestion in juvenile turot, Scophthlmus mximus nd effects of dietry dministrtion of Virio proteolyticus. Aquculture 186, 17-116. http://dx.doi.org/1.116/s44-8486(99)372-5. El-Ngr G, Clowes G, Tudoric CM nd Kuri V. 22. Rheologicl qulity nd stility of yog-ice crem with dded inulin. Int J Diry Technol 55, 89-93 http://dx.doi.org/146/ j.1471-37.22.42.x. Ersmus JH, Cook PA nd Coyne VE. 1997. The role of cteri in the digestion of seweed y the lone Hliotis mide. Aquculture 155, 377-386. http://dx.doi.org/1.116/ S44-8486(97)112-9. Glinski Z nd Jrosz J. 2. Immune phenomen in echinoderms. Arch Immunol Ther Exp (Wrsz) 48,189-193. Gison GR nd Roerfroid MB. 1995. Dietry modultion of the colonic microiot: Introducing the concept of preiotics. J Nutr 125, 141-1412. Gillilnd SE, Morelli L, Reid G. 21. Helth nd nutritionl properties of proiotics in food including powder milk with live lctic cid cteri. In: FAO/WHO Expert Consulttion On Evlution Of Helth And Nutritionl Properties of Proiotics in Food Including Powdered Milk. Cordo, Argentin. Gross PS, Al-Shrif WZ, Clow LA nd Smith LC. 1999. Echinoderm immunity nd the evolution of the complement system. Dev Comp Immunol 23, 429-442. Gu M, M H, Mi K, Zhng W, Bi N nd Wng X. 211. Effects of dietry β-glucn, mnnn oligoscchride nd their comintions on growth performnce, immunity nd resistnce ginst Virio splendidus of se cucumer Apostichopus jponicus. Fish Shellfish Immunol 31, 33-39. http:// dx.doi.org/1.116/j.fsi.211.5.18. Gu M, M HM, Mi KS, Zhng WB, Ai QH, Wng XJ nd Bi N. 21. Immune response of se cucumer Apostichopus jponicus coelomocytes to severl immunostimulnts in vitro. Aquculture 36, 49-56. http://dx.doi.org/1.116/j. quculture.21.5.24. Jing J Zhou Z, Dong Y, Gun X, Wng B, Jing B nd Sun H. 214. Chrcteriztion of phenoloxidse from the se cucumer Apostichopus jponicus. Immunoiology 219, 45-456. http://dx.doi.org/1.116/j.imio.214.2.6. Kn-no M nd Kijim A. 23. Genetic differentition mong three color vrints of Jpnese se cucumer Stichopus jponicus. Fish Sci 69, 86-812. http://dx.doi.org/1.146/ j.1444-296.23.69.x. Kng SA, Jng KH, Seo JW, Kim KH, Kim YH, Rirkhwd D nd Rhee SK. 29. Levn: pplictions nd perspectives. Microil production of iopolymers nd polymer precursors. Cister Acdemic Press, Norwich, 145-161. Kontul P, Suihko ML, Von Wright A nd Mttil-Sndholm T. 1999. The effect of lctose derivtives on intestinl lctic cid cteri. J Diry Sci 82, 249-256. http://dx.doi. org/1.3168/jds.s22-32(99)7523-6. Mgndóttir B, Gudmundsdottir BK, Lnge S, Steinrsson A, Oddgeirsson M, Bowden T nd Gudmundsdottir S. 26. Immunostimultion of lrve nd juveniles of cod, Gdus morhu. L J Fish Dis 29, 147-155. M Y, Liu Z, Yng Z, Li M, Liu J nd Song J. 213. Effects of dietry live yest Hnsenispor opuntie C21 on the immune nd disese resistnce ginst Virio splendidus infection in juvenile se cucumer Apostichopus jponicus. Fish Shellfish Immunol 34, 66-73. http://dx.doi.org/1.116/j. fsi.212.1.5. Morelli L, Zonenschin D, Cllegri ML, Grossi E, Misno F nd Fusillo M. 23. Assessment of new syniotic preprtion in helthy volunteers: survivl, persistence of proiotic strins nd its effect on the indigenousflor. Nutr J 2, 1-11. http://dx.doi.org/1.1186/1475-2891-2-11. Mourão PA, Pereir MS, Pvão MS, Mulloy B, Tollefsen DM, Mowinckel MC nd Aildgrd U. 1996. Structure nd nticogulnt ctivity of fucosylted chondroitin sulfte from echinoderm sulfted fucose rnches on the polyscchride ccount for its high nticogulnt ction. J Food Biochem 271, 23973-23984. Newj-Fyzul A, Al-Hri AH nd Austin B. 214. Review: developments in the use of proiotics for disese control in quculture. Aquculture 431, 1-11. http://dx.doi. org/1.116/j.quculture. 213.8.26. Nyk SK. 21. Proiotics nd immunity: fish perspective. Fish Shellfish Immunol 29, 2-14. http://dx.doi.org/1.116/j. fsi.21.2.17. Rmírez-Gómez F, Aponte-River F, Méndez-Cstner L nd Grcí-Arrrás J E. 21. Chnges in holothurin coelomocyte popultions following immune stimultion with different moleculr ptterns. Fish Shellfish Immunol 29, 175-185. http://dx.doi.org/1.116/j.fsi.21.3.13. Rmírez-Gómez F nd Grcí-Arrrás JE. 21. Echinoderm immunity. Invert Survivl J 7, 211-22. Rirkhwd D, Pl AK, Bhthen ZP, Shu NP, Jh A nd Mukherjee SC. 27. Dietry microil levn enhnces cellulr non-specific immunity nd survivl of common crp, Cyprinus crpio juveniles. Fish Shellfish Immunol 22, 477-486. http://dx.doi.org/1.116/j.fsi.26.6.5. Rehm BH. 29. Microil production of iopolymers nd polymer precursors: Applictions nd perspectives. Horizon Scientific Pres, Norfolk, UK. Srel M, Hllm K, Mttil-Sndholm T nd Mättö J. 23. The effect of lctose derivtives lctulose, lctitol nd lctoionic cid on the functionl nd technologicl properties of potentilly proiotic Lctocillus strins. Interntionl Diry J 13, 291-32. http://dx.doi.org/1.116/s958-6946(2)158-9. Sriknth R, Reddy CHS, Siddrth G, Rmih MJ nd Uppu-
홍해삼치삼의면역증강제섭이에따른영향 473 luri KB. 215. Review on production, chrcteriztion nd pplictions of microil levn. Crohydr Polym 12, 12-114. http://dx.doi.org/1.116/j.crpol.214.12.3. Surh S nd Shoo PK. 28. Lysozyme: n importnt defence molecule of fish innte immune system. Aquc Res 39, 223-239. http://dx.doi.org/1.1111/j.1365-219.27.1883.x. Shi C, Dong S, Wng F, Go Q nd Tin X. 213. Effects of four fresh microlge in diet on growth nd energy udget of juvenile se cucumer Apostichopus jponicus (Selenk). Aquculture 416, 296-31. http://dx.doi.org/1.116/j.quculture.213.9.5. Sugit H, Tkhshi J nd Deguchi Y. 1992. Production nd consumption of iotin y the intestinl microflor of cultured freshwter fishes. Biosci Biotechnol Biochem 56, 1678-1679. http://dx.doi.org/1.1271/.56.1678. Sun Y, Jin L, Wng T, Xue J, Liu G, Li X, You J, Li S nd Xu Y. 28. Polyscchrides from Astrglus memrnceuspromote phgocytosis nd superoxide nion (O 2 ) production y coelomocytes from se cucumer Apostichopus jponicus in vitro. Comp Biochem Physiol C 147, 293-298. http:// dx.doi.org/1.116/j.cpc.27.11.3. Ten Doeschte KI nd Coyne VE. 28. Improved growth rte in frmed Hliotis mide through proiotic tretment. Aquculture 284, 174-179. http://dx.doi.org/1.116/j.quculture.28.7.18. Tin Y, Ling XW, Chng YQ nd Song J. 215. Expression of c-type lysozyme gene in se cucumer Apostichopus jponicus is highly regulted nd time dependent fter slt stress. Comp Biochem Physiol B Biochem Mol Biol 18, 68-78. http://dx.doi.org/1.116/j.cp.214.1.4. Vine NG, Leukes WD nd Kiser H. 26. Proiotics in mrine lrviculture. FEMS Microiol Rev 3, 44-427. http:// dx.doi.org/1.1111/j.1574-6976.26.17.x. Wng F, Yng H, Go F, nd Liu G. 28. Effects of cute temperture or slinity stress on the immune response in se cucumer Apostichopus jponicus. Comp Biochem Physiol A Mol Integr Physiol 151, 491-498. http://dx.doi. org/1.116/j.cp.28.6.24. Wng X, Wng L, Che J, Li X, Li J, Wng J nd Xu Y. 214. In vitro non-specific immunostimultory effect of lginte oligoscchrides with different moleculr weights nd compositions on se cucumer Apostichopus jponicus coelomocytes. Aquculture 434, 434-441. http://dx.doi.org/1. 116/j.quculture.214.8.21. Xi S, Yng, H, Li Y, Liu S, Zhou Y nd Zhng L. 212. Effects of different seweed diets on growth, digestiility, nd mmoni-nitrogen production of the se cucumer Apostichopus jponicus (Selenk). Aquculture 338, 34-38. http:// dx.doi.org/1.116/j.quculture.212.1.1. Xue Z, Li H, Wng X, Li X, Liu Y, Sun J nd Liu C. 215. A review of the immune molecules in the se cucumer. Fish Shellfish Immunol 44, 1-11. http://dx.doi.org/1.116/j. fsi.215.1.26. Yng H, Yun X, Zhou Y, Mo Y, Zhng T nd Liu, Y. 25. Effects of ody size nd wter temperture on food consumption nd growth in the se cucumer Apostichopus jponicus (Selenk) with specil reference to estivtion. Aquc Res 36, 185-192. http://dx.doi.org/1.1111/j.1365-219.25.1325.x. Yu MC, Li ZJ, Lin HZ, Wen GL nd M S. 29. Effects of dietry medicinl hers nd Bcillus on survivl, growth, ody composition, nd digestive enzyme ctivity of the white shrimp, Litopeneus vnnmei. Aquc Int 17, 377-384. http://dx.doi.org/1.17/s1499-8-929-3. Yu L, Xue C, Chng Y, Hu Y, Xu X, Ge L nd Liu G. 215. Structure nd rheologicl chrcteristics of fucoidn from se cucumer Apostichopus jponicus. Food Chem 18, 71-76. http://dx.doi.org/1.116/j.foodchem.215.2.34. Zho W, Ling M nd Zhng P. 21. Effect of yest polyscchride on the immune function of juvenile se cucumer Apostichopus jponicus (Selenk) under ph stress. Aqucult Int 18, 777-786. http://dx.doi.org/1.17/s1499-9- 93-4. Zokeifr H, Blcázr JL, Sd CR, Kmrudin MS, Sijm K, Arshd A nd Nejt N. 212. Effects of Bcillus sutilis on the growth performnce, digestive enzymes, immune gene expression nd disese resistnce of white shrimp, Litopeneus vnnmei. Fish Shellfish Immunol 33, 683-689. http://dx.doi.org/1.116/j.fsi.212.5.27.