ankara univ vet fak derg Ankara Üniversitesi Veteriner Fakültesi Dergisi 1300-0861 1308-2817 Ankara University 10.1501/Vetfak_0000002863 Veterinary Surgery Veteriner Cerrahi Immune response and disease resistance in the white shrimp, Litopenaeus vannamei induced by potential probiotic Lactobacillus Immune response and disease resistance in the white shrimp, Litopenaeus vannamei induced by potential probiotic Lactobacillus Roomıanı Laleh Ahmadı Sara Ghaenı Mansoreh 09 01 2018 65 3 323 329 09 01 2018 Copyright © 1954, Ankara Üniversitesi Veteriner Fakültesi Dergisi 1954 Ankara Üniversitesi Veteriner Fakültesi Dergisi

In the present study, Lactobacillus bulgaricus E20 isolated from the gut contents of farm-reared white shrimp, Litopenaeus vannamei, after determination of its characteristics using biochemical, molecular and probiotical examinations, was used in the diet of white shrimp to evaluate the survival rate, immune status, and disease resistance. L. bulgaricus was administered at two different doses 1×107 (T1) and 1×109 (T2) CFU g_1 feed to shrimp for 30 days. A control group was also included with normal feed. The survival rate, total hemocyte counts, phenoloxidase activity, respiratory burst, gut lactic acid bacteria and gut total bacteria were evaluated at the end of trial and after challenging shrimp with Vibrio parahaemolyticus. Higher survival rates were observed in shrimp fed probiotic diets as compared with the control (P<0.05). The best immune performance in terms of hemocyte counts, phenoloxidase activity, and respiratory burst was seen in the probiotic-treated groups especially in T2. Also, an improvement was seen in the number of lactic acid bacterias in shrimp that had been given probiotics. There were no significant differences between groups (treated and control) after exposing with V. parahemolyticus (P<0.05). Shrimp fed with probiotic diets revealed lower cumulate mortality than the control group. These findings demonstrated that administration of L. bulgaricus can improve survival rate and disease resistance through an enhanced immune response in shrimp

In the present study, Lactobacillus bulgaricus E20 isolated from the gut contents of farm-reared white shrimp, Litopenaeus vannamei, after determination of its characteristics using biochemical, molecular and probiotical examinations, was used in the diet of white shrimp to evaluate the survival rate, immune status, and disease resistance. L. bulgaricus was administered at two different doses 1×107 (T1) and 1×109 (T2) CFU g_1 feed to shrimp for 30 days. A control group was also included with normal feed. The survival rate, total hemocyte counts, phenoloxidase activity, respiratory burst, gut lactic acid bacteria and gut total bacteria were evaluated at the end of trial and after challenging shrimp with Vibrio parahaemolyticus. Higher survival rates were observed in shrimp fed probiotic diets as compared with the control (P<0.05). The best immune performance in terms of hemocyte counts, phenoloxidase activity, and respiratory burst was seen in the probiotic-treated groups especially in T2. Also, an improvement was seen in the number of lactic acid bacterias in shrimp that had been given probiotics. There were no significant differences between groups (treated and control) after exposing with V. parahemolyticus (P<0.05). Shrimp fed with probiotic diets revealed lower cumulate mortality than the control group. These findings demonstrated that administration of L. bulgaricus can improve survival rate and disease resistance through an enhanced immune response in shrimp

 Immune responses Lactobacillus bulgaricus Litopenaeus vannamei probiotic Immune Litopenaeus Lactobacillus Alishahi M, Ranjbar MM, Ghorbanpour M, et al. (2010): Effects of dietary Aloe vera on specific and nonspecific immunity of Common carp (Cyprinus carpio). J Vet Res, 4, 85-91. Amann RI, Ludwig W, Schleifer KH (1995): Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev, 59, 143-169. Amparyup P, Charoensapsri W, Tassanakajon A (2013): Prophenoloxidase system and its role in shrimp immune responses against major pathogens. Fish Shell Immunol, 34, 990-1001. Anderson DP, Siwicki AK (1995): Basic haematology and serology for fish health program. 185- 202. In: M Shariff, JR Arthur, RP Subasinghe (eds.) Fish Health Section. Asian Fisheries Society, Manila, Phillipines. Argue BJ, Arce SM, Lotz JM, et al. (2002): Selective breeding of Pacific white shrimp (Litopenaeus vannamei) for growth and resistance to Taura syndrome virus. Aquaculture, 204, 447-460. Bradford MM (1976): Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 72, 248- 254. Chiu CH, Guu YK, Liu CH, et al. (2007): Immune responses and gene expression in white shrimp, Litopenaeus vannamei, induced by Lactobacillus plantarum. Fish Shell Immunol, 23, 364-377. Farzanfar A (2006): The use of probiotics in shrimp aquaculture. FEMS Immunol Med Microbiol, 48, 149-158. Febrianti D, Yuhana M, Widanarni A (2016): Dietary synbiotic microcapsule influences the immune responses, growth performance and microbial populations to white spot syndrome virus in pacific white shrimp (Litopenaeus vannamei). J Fish Aquat Sci, 11, 28-42. Graslund S, Karin K, Wongtavatchai J (2002): Responsible use of antibiotic in shrimp farming. Aquacult Asia, 7, 17. Hao K, Liu JY, Ling F, et al. (2014): Effects of dietary administration of Shewanella haliotis D4, Bacillus cereus D7 and Aeromonas bivalvium D15, single or combined, on the growth, innate immunity and disease resistance of shrimp, Litopenaeus vannamei. Aquaculture, 428, 141-149. Holt J, Krieg N, Sneath P (1984): Bergey’s Manual of Systematic Bacteriology. Vol. 1. The Williams and Wilkins Co, Baltimore. Irianto A, Austin B (2002): Use of probiotics to control furunculosis in rainbow trout, Oncorhynchus mykiss (Walbaum). J Fish Dis, 25, 333-342. Jayanth K, Jeyasekaran G, Jeya Shakila R (2001): Biocontrol of fish bacterial pathogens by the antagonistic bacteria isolated from the Coastal Waters of Gulf of Mannar, India. Bull Eur Assoc Fish Pathol, 21, 1218. Li P, Burr GS, Gatlin DM, et al. (2007): Dietary supplementation of short-chain fructo-oligosaccharides influences gastrointestinal microbiota composition and immunity characteristics of Pacific white shrimp, Litopenaeus vannamei, cultured in a recirculating system. J Nutr, 137, 2763-68. Li J, Tan B, Mai K, et al. (2008): Immune responses and resistance against Vibrio parahaemolyticus induced by probiotic bacterium Arthrobacter XE-7 in Pacific white shrimp, Litopenaeus vannamei. J World Aquacult Soc, 39, 477-489. Li J, Tan B, Mai K (2009): Dietary probiotic Bacillus OJ and isomaltooligosaccharides influence the intestine microbial populations, immune responses and resistance to white spot syndrome virus in shrimp (Litopenaeus vannamei). Aquaculture, 291, 35-40. Liu CH, Chiu CS, Lin PL, et al. (2009): Improvement in the growth performance of white shrimp, Litopenaeus vannamei, by a protease producing probiotic, Bacillus subtilis E20 from natto. J Appl Microbiol, 107, 1031-41. McIntyre J, Choonara I (2004): Drug toxicity in the neonate. Biol Neonate, 86, 218-221. Merrifield DL, Dimitroglou A, Foey A, et al. (2010): The current status and future focus of probiotic and prebiotic applications for salmonids. Aquaculture, 302, 1-18. Nikoskelainen S, Ouwehand AC, Bylund G, et al. (2003): Immune enhancement in rainbow trout (Oncorhynchus mykiss) by potential probiotic bacteria (Lactobacillus rhamnosus). Fish Shell Immunol, 15, 443-452. Nurhayati D, Widanarni A, Yuhana M (2015): Dietary synbiotic influence on the growth performances and immune responses to co-infection with infectious myonecrosis virus and Vibrio harveyi in Litopenaeus vannamei. J Fish Aqua Sci, 10, 13-23. Panigrahi A, Kiron V, Satoh S, et al. (2007): Immune modulation and expression of cytokine genes in rainbow trout Oncorhynchus mykiss upon probiotic feeding. Dev Comp Immunol, 31, 372-382. Pantoja CR, Song X, Xia L, et al. (2005): Development of a specific pathogen-free (SPF) population of the Chinese fleshy prawn Fenneropenaeus chinensis Part 1: disease pre-screening and primary quarantine. Aquaculture, 250, 573-578. Planas M, Vázquez JA, Marqués J, et al. (2004): Enhancement of rotifer (Brachionus plicatilis) growth by using terrestrial lactic acid bacteria. Aquaculture, 240, 313-329. Reed LA, Siewicki TC, Shah AC (2006): The biopharmaceutics and oral bioavailability of two forms of oxytetracycline to the white shrimp, Litopenaeus setiferus. Aquaculture, 258, 42-54. Rengpipat S, Rukpratanporn S, Piyatiratitivorakul S, et al. (2000): Immunity enhancement in black tiger shrimp (Penaeus monodon) by a probiont bacterium (Bacillus S11). Aquaculture, 191, 271-288. Ringo E, Olsen RE, Gifstad TO, et al. (2010): Prebiotics in aquaculture: A review. Aquacult Nutr, 16, 117-136. Rodriguez J, Le Moullac G (2000): State of the art of immunological tools and health control of penaeid shrimp. Aquaculture, 191, 109-119. Ruiz C, Roman G, Sanchez J (1996): A marine bacterial strain effective in producing antagonisms of other bacteria. Aquac Int, 4, 289-291. Sahu MK, Swarnakumar NS, Sivakumar K, et al. (2008): Probiotics in aquaculture: Importance and future perspectives. Ind J Microbiol, 48, 299-308. Saurabh S, Sahoo PK (2008): Lysozyme: An important defense molecule of fish innate immune system. Aquacult Res, 39, 223-239. Singh SK, Tiwari VK, Chadha NK, et al. (2013): Effect of supplementation on growth and haemato-immunological function of Labeo rohita (Hamilton, 1822) fingerlings exposed to sub-lethal nitrite stress. Israeli J Aquacult - Bamidgeh, 64, 1-11. and fructooligosaccharide Sivakumar N, Sundararaman M, Selvakumar G (2012): Probiotic effect of Lactobacillus acidophilus against vibriosis in juvenile shrimp (Penaeus monodon). Afr J Biotechnol, 11, 15811-18. Smith JV, Soderhall K (1983): Induction of degranulation and lysis of haemocytes in the fresh water crayfish, Astacus astacus by components of the prophenoloxidase activating system in vitro. Cell Tissue Res, 233, 295-303. Soto-Rodriguez S, Armenta M, Gomez-Gil B (2006): Effects of enrofloxacin and florfenicol on survival and bacterial population in an experimental infection with luminescent Vibrio campbellii in shrimp larvae of Litopenaeus vannamei. Aquaculture, 255, 48-54. Sung HH, Yang YL, Song YL (1996): Enhancement of microbicidal activity in the tiger shrimp Penaeus monodon via immunostimulation. J Crustacean Biol, 16, 278-284. Tendencia EA, De la Peña LD (2002): Level and percentage recovery of resistance to oxytetracycline and oxolinic acid of bacteria from shrimp ponds. Aquaculture, 213, 1-13. Tseng DY, Ho PL, Huang SY, et al. (2009): Enhancement of immunity and disease resistance in the white shrimp, Litopenaeus vannamei, by the probiotic, Bacillus subtilis E20. Fish Shell Immunol, 26, 339-344. Vargas-Albores F, Gşzman MA, Ochoa JL (1993): An anticoagulant solution for haemolymph collection and prophenoloxidase studies of penaeid shrimp (Penaeus californiensis). Comp Biochem Physiol, 106A, 299-303. Verschuere L, Rombaut G, Sorgeloos P, et al. (2000): Probiotic bacteria as biological control agents in aquaculture. Microbiol Mol Biol Rev, 64, 655-671. Wang Y (2007): Effect of probiotics on growth performance and digestive enzyme activity of the shrimp Penaeus vannamei. Aquaculture, 269, 259-264. You XX, Su YQ, Mao Y, et al. (2010): Effect of high water temperature on mortality, immune response and viral replication of WSSV-infected Marsupenaeus japonicas juveniles and adults. Aquaculture, 305, 133-137. Ziaei-Nejad S, Rezaei MH, Takami GA, et al. (2006): The effect of Bacillus spp. bacteria used as probiotics on digestive enzyme activity, survival and growth in the India white shrimp Fenneropenaeus indicus. Aquaculture, 252, 516-24.