Effects of dietary butyrate monoglyceride and probiotic of Bacillus on growth performance and enzymatic antioxidant activity of Nile tilapia (Oreochromis niloticus)

Document Type : Research Paper

Authors

1 M.Sc. in Aquatic Breeding, Department of Fisheries, Faculty of Nano and Bioscience and Technology, Persian Gulf University, Bushehr, Iran

2 Assistant Professor in Department of Fisheries, Faculty of Nano and Bioscience and Technology, Persian Gulf University, Bushehr, Iran

3 Assistant Professor in Department of Fisheries and Biology, Persian Gulf Research Institute, Persian Gulf University, Bushehr, Iran

10.22124/japb.2022.22091.1461

Abstract

In this study, the effect of dietary butyrate monoglyceride and probiotic (Bacillus subtilis and Bacillus licheniformis) supplementation were evaluated on growth performance and the enzymatic antioxidant activity of Nile tilapia (Oreochromis niloticus). Fish were divided into 4 treatments with 3 replications including control, 10g/kg butyrate monoglyceride (AB1), 5g/kg probiotic Bacillus (AB2), 10g/kg butyrate monoglyceride and 5g/kg probiotic Bacillus (AB3). 144 fish with an average weight of 12±0.58g were fed for 8 weeks. This study showed that applying butyrate monoglyceride and probiotic Bacillus in the feeding of Nile tilapia significantly improved the growth indices of final weight gain (FWG), daily weight gain (DWG), protein efficiency ratio (PER), and FCR compared to control (P<0.05). All experimental diets had higher levels of catalase, glutathione peroxidase and superoxide dismutase activity compared to control (P<0.05). However, the highest levels of superoxide dismutase activity were measured in treatment AB3. The present study indicated that singular and combined dietary administration of butyrate monoglyceride and probiotic Bacillus had a significant impact on growth performance, and liver and serum enzymatic antioxidant activity of Nile tilapia.

Keywords

Main Subjects


بادزهره غ.، زارعی ش.،  داوودی ر.،  نفیسی بهابادی م.، صالحی ف.، مرشدی و. و ستوده ا. 1399. تاثیر سطوح مختلف اسید بوتیریک در جیره غذایی بر برخی عملکرد رشد، ایمنی و فعالیت آنزیم­های گوارشی ماهی شانک زرد باله (Acanthopagrus latus Houttuyn, 1782). تغذیه آبزیان، 6(3): 67-55.
خالقی م.، سلطانی م. و حسینی شکرابی س.پ. 1397. اثر افزودن مخلوط پروبیوتیک Bacillus subtilis و Bacillus licheniformis  (دی‌پرو آکوا) به جیره غذایی بر برخی شاخص‌های رشد، خونی و ایمنی ماهی کپور معمولی. زیست‌شناسی دریا، 10(38):           20-11.
کامگار م.، قانع م.، پورغلام ر. و قیاسی م. 1391. تاثیر Bacillus subtilis به عنوان پروبیوتیک بر فاکتورهای هماتولوژی و بیوشیمیایی خون ماهی قزل‌آلای رنگین‌کمان (Oncorhynchus mykiss) به دنبال عفونت تجربی با Streptococcus iniae. توسعه آبزی‌پروری، 6(1): 102-91.
مازندرانی م.، سوداگر م.، جعفری و.، جافرنوده ع. و بزی ف. 1396. اثرات تغذیه‌ای اسید بوتیریک خوراکی در ماهی کلمه خزری (Rutilus caspicus) در مواجهه با استرس شوری. آبزیان زینتی، ۴(۳): ۴۰-۳۱.
Aalamifar H., Soltanian S., Vazirzadeh A., Akhlaghi M., Morshedi V., Gholamhosseini A. and Torfi Mozanzadeh M. 2020. Dietary butyric acid improved growth, digestive enzyme activities and humoral immune parameters in Barramundi (Lates calcarifer). Aquacultre Nutrition, 26: 156–164.
Abarike E.D., Cai J., Lu Y., Yu H., Chen L., Jian J., Jian J., Tang J., Jun J. and Kuebutornye F.K.A. 2018. Effects of a commercial probiotic BS containing Bacillus subtilis and Bacillus licheniformis on growth, immune response and disease resistance in Nile tilapia, Oreochromis niloticus. Fish and Shellfish Immunology, 82: 229–238.
Abd El-Gawad E.A., Abd El-Latif A.M. and Shourbela R.M. 2016. Enhancement of antioxidant activity, non-specific immunity and growth performance of Nile tilapia, Oreochromis niloticus by dietary fructooligosaccharide. Journal of Aquaculture Research and Development, 7(5): 1–7 (427).
Abd El-Naby A.S., Khattaby A.E.R.A., Samir F., Awad S.M.M. and Abdel-Tawwab M. 2019. Stimulatory effect of dietary butyrate on growth, immune response, and resistance of Nile tilapia, Oreochromis niloticus against Aeromonas hydrophila infection. Animal Feed Science and Technology, 254: 1–9 (114212).
Abdel-Latif H.M.R., Abdel-Tawwab M., Dawood M.A.O., Menanteau-Ledouble S. and El-Matbouli M. 2020. Benefits of dietary butyric acid, sodium butyrate, and their protected forms in aquafeeds: A review. Reviews in Fisheries Science and Aquaculture, 28: 421–448.
Abriouel H., Franz C.M.A.P., Omar N.B. and Galvez A. 2011. Diversity and applications of Bacillus bacteriocins. FEMS Microbiology Reviews, 35(1): 201–232.
Abu Elala N.M. and Ragaa N.M. 2015. Eubiotic effect of a dietary acidifier (potassium diformate) on the health status of cultured Oreochromis niloticus. Journal of Advanced Research, 6(4): 621–629.
Aebi H. 1984. Catalase in vitro. Methods in Enzymology, 105: 121–126.
Ahmed H. and Mohamed Sadek K. 2014. Impact of dietary supplementation of sodium butyrate and/or protexin on the growth performance, some blood parameters, and immune response of Oreochromis niloticus. International Journal of Agriculture Innovations and Research, 3(4): 985–991.
Bedford A. and Gong J. 2018. Implications of butyrate and its derivatives for gut health and animal production. Animal Nutrition, 4:151–159.
Benedito-Palos L., Ballester-Lozano G.F., Simo P., Karalazos V., Ortiz A., Calduch-Giner J. and Perez-Sanchez J. 2016. Lasting effects of butyrate and low FM/FO diets on growth performance, blood hematology/biochemistry and molecular growth-related markers in gilthead sea bream (Sparus aurata). Aquaculture, 454: 8–18.
Brunt J., Newaj-Fyzul A. and Austin B. 2007. The development of probiotics for the control of multiple bacterial diseases of rainbow trout, Oncorhynchus mykiss (Walbaum). Journal of Fish Diseases, 30(10): 573–579.
Castillo S., Rosales M., Pohlenz C. and Gatlin D.M. 2014. Effects of organic acids on growth performance and digestive enzyme activities of juvenile red drum Sciaenops ocellatus. Aquaculture, 433: 6–12.
Da Silva B.C., Do Vieira F.N., Mourino J.L.P., Bolivar N. and Seiffert W.Q. 2016. Butyrate and propionate improve the growth performance of Litopenaeus vannamei. Aquaculture Research, 47(2): 612–623.
Dai J., Li Y., Yang P., Liu Y., Chen Z., Ou W., Ai Q., Zhang W., Zhang Y. and Mai K. 2018. Citric acid as a functional supplement in diets for juvenile turbot, Scophthalmus maximus L.: Effects on phosphorus discharge, growth performance, and intestinal health. Aquaculture, 495: 643–653.
Darafsh F., Soltani M., Abdolhay H.A. and Shamsaei Mehrejan M. 2020. Improvement of growth performance, digestive enzymes and body composition of Persian sturgeon (Acipenser persicus) following feeding on probiotics: Bacillus licheniformis, Bacillus subtilis and Saccharomyces cerevisiae. Aquaculture Research, 51(3): 957–964.
Dawood M.A.O., Eweedah N.M., Elbialy Z.I. and Abdelhamid A.I. 2020. Dietary sodium butyrate ameliorated the blood stress biomarkers, heat shock proteins, and immune response of Nile tilapia (Oreochromis niloticus) exposed to heat stress. Journal of Thermal Biology, 88: 1–10 (102500).
Di J., Chu Z., Zhang S., Huang J., Du H. and Wei Q. 2019. Evaluation of the potential probiotic Bacillus subtilis isolated from two ancient sturgeons on growth performance, serum immunity and disease resistance of Acipenser dabryanus. Fish and Shellfish Immunology, 93: 711–719.
El-Haroun E.R., Goda A.M.A.S. and Kabir Chowdhury M.A. 2006. Effect of dietary probiotic Biogen supplementation as a growth promoter on growth performance and feed utilization of Nile tilapia Oreochromis niloticus (L.). Aquaculture Research, 37(14): 1473–1480.
FAO (Food and Agricultural Organization) 2017. FAO Yearbook: Statistics Fisheries and Aquaculture Statistics 2017/FAO Annuaire. FAO, Italy. 107P.
Franke A., Roth O., De Schryver P., Bayer T., Garcia-Gonzalez L., Kunzel S., Bossier P., Miest J.J. and Clemmesen C. 2017. Poly-β-hydroxybutyrate administration during early life: Effects on performance, immunity and microbial community of European sea bass yolk-sac larvae. Scientific Reports, 7(1): 1–11 (15022).
Fuller R. 1989. Probiotics in man and animals. Journal of Applied Bacteriology, 66: 365–378.
Han B., Long W.Q., He J.Y., Liu Y.J., Si Y.Q. and Tian L.X. 2015. Effects of dietary Bacillus licheniformis on growth performance, immunological parameters, intestinal morphology and resistance of juvenile Nile tilapia (Oreochromis niloticus) to challenge infections. Fish and Shellfish Immunology, 46(2): 225–231.
Hassaan M.S., Soltan M.A., Jarmołowicz S. and Abdo H.S. 2018. Combined effects of dietary malic acid and Bacillus subtilis on growth, gut microbiota and blood parameters of Nile tilapia (Oreochromis niloticus). Aquaculture Nutrition, 24(1): 83–93.
He S., Zhang Y., Xu L., Yang Y., Marubashi T., Zhou Z. and Yao B. 2013. Effects of dietary Bacillus subtilis C-3102 on the production, intestinal cytokine expression and autochthonous bacteria of hybrid tilapia Oreochromis niloticus ♂ × Oreochromis aureus ♀. Aquaculture, 412-413: 125–130.
Hoseinifar S.H., Sun Y.Z. and Caipang C.M. 2017. Short-chain fatty acids as feed supplements for sustainable aquaculture: An updated view. Aquaculture Research, 48: 1380–1391.
Hussein M.S., Zaghlol A., El-Hakim N.F.A., El Nawsany M. and Abo-State H.A. 2016. Effect of different growth promoters on growth performance, feed utilization and body composition of common carp (Cyprinus carpio). Journal of Fisheries and Aquatic Science, 11(5): 370–377.
Jahanian R. and Golshadi M. 2015. Effect of dietary supplementation of butyric acid glycerides on performance, immunological responses, ileal microflora, and nutrient digestibility in laying hens fed different basal diets. Livestock Science, 178: 228–236.
Jesus G.F.A., Pereira S.A., Owatari M.S., Syracuse N., Silva B.C., Silva A., Pierr B.S., Lehmann N.B., Figueired H.C.P., Fracalossi D.M., Mourino J.L.P. and Martins M.L. 2019. Protected forms of sodium butyrate improve the growth and health of Nile tilapia fingerlings during sexual reversion. Aquaculture, 499: 119–127.
John G., Mohamed R.R., Kolanchina P. and Balasundar A. 2018. Nutritional value of two bacterial strains Bacillus subtilis RCMB21 and Pseudomonas fluorescens RCMB39 as feed supplement for freshwater prawn Macrobrachium malcolmsonii. Asian Journal of Animal and Veterinary Advances, 13(3): 282–294.
Kesarcodi-Watson A., Kaspar H., Lategan M.J. and Gibson L. 2008. Probiotics in aquaculture: The need, principles and mechanisms of action and screening processes. Aquaculture, 274(1): 1–14.
Kumar P., Jain K.K. and Sardar P. 2018. Effects of dietary synbiotic on innate immunity, antioxidant activity and disease resistance of Cirrhinus mrigala juveniles. Fish and Shellfish Immunology, 80: 124–132.
Kuthan H., Haussmann H.J. and Werringloer J. 1986. A spectrophotometric assay for superoxide dismutase activities in crude tissue fractions. Biochemical Journal, 237(1): 175–180.
Lee B.J., Kim S.S., Song J.W., Oh D.H., Cha J.H., Jeong J.B., Heo M.S., Kim K.W. and Lee K.J. 2013. Effects of dietary supplementation of citrus by-products fermented with a probiotic microbe on growth performance, innate immunity and disease resistance against Edwardsiella tarda in juvenile olive flounder, Paralichthys olivaceus (Temminck & Schlegel). Journal of Fish Diseases, 36(7): 617–628.
Li W.F., Deng B., Cui Z.W., Fu L.Q., Chen N.N., Zhou X.X., Shen W.Y. and Yu D.Y. 2012. Several indicators of immunity and antioxidant Activities improved in grass carp given a diet containing Bacillus additive. Journal of Animal and Veterinary Advances, 11: 2392–2397.
Lin Y.H. and Cheng M.Y. 2017. Effects of dietary organic acid supplementation on the growth, nutrient digestibility and intestinal histology of the giant grouper Epinephelus lanceolatus fed a diet with soybean meal. Aquaculture, 469: 106–111.
Liu C.H., Chiu C.H., Wang S.W. and Cheng W. 2012. Dietary administration of the probiotic, Bacillus subtilis E20, enhances the growth, innate immune responses, and disease resistance of the grouper, Epinephelus coioides. Fish and Shellfish Immunology, 33: 699–706.
Liu M., Guo W., Wu F., Qu Q., Tan Q. and Gong W. 2017. Dietary supplementation of sodium butyrate may benefit growth performance and intestinal function in juvenile grass carp (Ctenopharyngodon idellus). Aquaculture Research, 48(8): 4102–4111.
Liu W., Yang Y., Zhang J., Gatlin D.M., Ringo E. and Zhou Z. 2014. Effects of dietary microencapsulated sodium butyrate on growth, intestinal mucosal morphology, immune response and adhesive bacteria in juvenile common carp (Cyprinus carpio) pre-fed with or without oxidized oil. British Journal of Nutrition, 112(1): 15–29.
Luckstadt C. 2008. The use of acidifiers in fish nutrition. CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources, 3: 1–8 (044).
Luo F., Fu Z., Wang M., Ke Z., Wang M., Wang W., Hassan M. and Shu X. 2021. Growth performance, tissue mineralization, antioxidant activity and immune response of Oreochromis niloticus fed with conventional and gluconic acid zinc dietary supplements. Aquaculture Nutrition, 27(3): 897–907.
Martinez-Alvarez R.M., Morales A.E. and Sanz A. 2005. Antioxidant defenses in fish: Biotic and abiotic factors. Reviews in Fish Biology and Fisheries, 15(1-2): 75–88.
Mirghaed A.T., Yarahmadi P., Soltani M., Paknejad H. and Hoseini S.M. 2019. Dietary sodium butyrate (Butirex® C4) supplementation modulates intestinal transcriptomic responses and augments disease resistance of rainbow trout (Oncorhynchus mykiss). Fish and Shellfish Immunology, 92: 621–628.
Mohapatra S., Chakraborty T., Kumar V., DeBoeck G. and Mohanta K.N. 2013. Aquaculture and stress management: A review of probiotic intervention. Journal of Animal Physiology and Animal Nutrition, 97(3): 405–430.
Morken T., Kraugerud O.F., Barrows F.T., Sorensen M., Storebakken T. and Overland M. 2011. Sodium diformate and extrusion temperature affect nutrient digestibility and physical quality of diets with fish meal and barley protein concentrate for rainbow trout (Oncorhynchus mykiss). Aquaculture, 317(1-4): 138–145.
Naderi Farsani M., Bahrami Gorji S., Hoseinifar S.H., Rashidian G. and Van Doan H. 2020. Combined and singular effects of dietary PrimaLac® and potassium diformate (KDF) on growth performance and some physiological parameters of rainbow trout (Oncorhynchus mykiss). Probiotics and Antimicrobial Proteins, 12(1): 236–245.
Najdegerami E.H., Tran T.N., Defoirdt T., Marzorati M., Sorgeloos P., Boon N. and Bossier P. 2012. Effects of poly-β-hydroxybutyrate (PHB) on Siberian sturgeon (Acipenser baerii) fingerlings performance and its gastrointestinal tract microbial community. FEMS Microbiology Ecology, 79(1): 25–33.
Ng W.K. and Koh C.B. 2017. The utilization and mode of action of organic acids in the feeds of cultured aquatic animals. Reviews in Aquaculture, 9(4): 342–368.
Olmos Soto J. 2017. Bacillus probiotics enzymes: External auxiliary apparatus to avoid digestive deficiencies, water pollution, diseases, and economic problems in marine cultivated animals. Advances in Food and Nutrition Research, 80: 15–35.
Omosowone O.O., Dada A.A. and Adeparusi E.O. 2018. Comparison of dietary butyric acid supplementation effect on growth performance and body composition of Clarias gariepinus and Oreochromis niloticus fingerlings. Iranian Journal of Fisheries Sciences, 17(2): 403–412.
Park Y., Kim H., Won S., Hamidoghli A., Hasan M.T., Kong I. and Bai S.C. 2020. Effects of two dietary probiotics (Bacillus subtilis or licheniformis) with two prebiotics (mannan or fructo-oligosaccharide) in Japanese eel, Anguilla japonica. Aquaculture Nutrition, 26(2): 316–327.
Peixoto M.J., Salas-Leiton E., Pereira L.F., Queiroz A., Magalhaes F., Pereira R., Abreu H., Reis P.A., Goncalves J.F.M. and De Almeida Ozorio R.O. 2016. Role of dietary seaweed supplementation on growth performance, digestive capacity and immune and stress responsiveness in European seabass (Dicentrarchus labrax). Aquaculture Reports, 3: 189–197.
Reda R.M., Mahmoud R., Selim K.M. and El-Araby I.E. 2016. Effects of dietary acidifiers on growth, hematology, immune response and disease resistance of Nile tilapia, Oreochromis niloticus. Fish and Shellfish Immunology, 50: 255–262.
Rimoldi S., Finzi G., Ceccotti C., Girardello R., Grimaldi A., Ascione C. and Terova G. 2016. Butyrate and taurine exert a mitigating effect on the inflamed distal intestine of European sea bass fed with a high percentage of soybean meal. Fisheries and Aquatic Sciences, 19(1): 1–14 (40).
Robles R., Lozano A.B., Sevilla A., Marquez L., Nuez-Ortin W. and Moyano F.J. 2013. Effect of partially protected butyrate used as feed additive on growth and intestinal metabolism in sea bream (Sparus aurata). Fish Physiology and Biochemistry, 39(6): 1567–1580.
Soltani M., Ghosh K., Hoseinifar S.H., Kumar V., Lymbery A.J., Roy S. and Ringo E. 2019. Genus Bacillus, promising probiotics in aquaculture: Aquatic animal     origin, bio-active components, bioremediation and efficacy in fish and shellfish. Reviews in Fisheries Science and Aquaculture, 27(3): 331–379.
Sun Y.Z., Yang H.L., Ma R.L. and Lin W.Y. 2010. Probiotic applications of two dominant gut Bacillus strains with antagonistic activity improved the growth performance and immune responses of grouper Epinephelus coioides. Fish and Shellfish Immunology, 29(5): 803–809.
Tang L., Huang K., Xie J., Yu D., Sun L., Huang Q. and Bi Y. 2017. 1-Deoxynojirimycin from Bacillus subtilis improves antioxidant and antibacterial activities of juvenile Yoshitomi tilapia. Electronic Journal of Biotechnology, 30: 39–47.
Terova G., Diaz N., Rimoldi S., Ceccotti C., Gliozheni E. and Piferrer F. 2016. Effects of sodium butyrate treatment on histone modifications and the expression of genes related to epigenetic regulatory mechanisms and immune response in European sea bass (Dicentrarchus labrax) fed a plant-based diet. PLOS One, 11(7): 1–20 (0160332).
Tseng D.Y., Ho P.L., Huang S.Y., Cheng S.C., Shiu Y.L., Chiu C.S. and Liu C.H. 2009. Enhancement of immunity and disease resistance in the white shrimp, Litopenaeus vannamei, by the probiotic, Bacillus subtilis E20. Fish and Shellfish Immunology, 26(2): 339–344.
Vidhya Hindu S., Thanigaivel S., Vijayakumar S., Chandrasekaran N., Mukherjee A. and Thomas J. 2018. Effect of microencapsulated probiotic Bacillus vireti 01-polysaccharide extract of Gracilaria folifera with alginate-chitosan on immunity, antioxidant activity and disease resistance of Macrobrachium rosenbergii against Aeromonas hydrophila infection. Fish and Shellfish Immunology, 73: 112–120.
Volatiana J.A., Sagada G., Xu B., Zhang J., Ng W. and Shao Q. 2020. Effects of butyrate glycerides supplementation in high soybean meal diet on growth performance, intestinal morphology and antioxidative status of juvenile black sea bream, Acanthopagrus schlegelii. Aquaculture Nutrition, 26(1): 15–25.
Wang Y., Wu Y., Wang Y., Xu H., Mei X., Yu D., Wang Y. and Li W. 2017. Antioxidant properties of probiotic bacteria. Nutrients, 9(5): 1–15 (9050521).
Wang Y.B., Li J.R. and Lin J. 2008. Probiotics in aquaculture: Challenges and outlook. Aquaculture, 281: 1–4.
Weifen L., Xiaoping Z., Wenhui S., Bin D., Quan L., Luoqin F., Jiajia J., Yue W. and Dongyou Y. 2012. Effects of Bacillus preparations on immunity and antioxidant activities in grass carp (Ctenopharyngodon idellus). Fish Physiology and Biochemistry, 38(6): 1585–1592.
Won S., Hamidoghli A., Choi W., Bae J., Jang W.J., Lee S. and Bai S.C. 2020. Evaluation of potential probiotics Bacillus subtilis WB60, Pediococcus pentosaceus, and Lactococcus lactis on growth performance, immune response, gut histology and immune-related genes in whiteleg shrimp, Litopenaeus vannamei. Microorganisms, 8(2): 1–15 (8020281).
Wu P., Tian L., Zhou X.Q., Jiang W.D., Liu Y., Jiang J., Xie F., Kuang S.Y., Tang L., Tang W.N., Yang J., Zhang Y.A., Shi H.Q. and Feng L. 2018. Sodium butyrate enhanced physical barrier function referring to Nrf2, JNK and MLCK signaling pathways in the intestine of young grass carp (Ctenopharyngodon idella). Fish and Shellfish Immunology, 73: 121–132.
Yang G., Cao H., Jiang W., Hu B., Jian S., Wen C., Kajbaf K., Kumar V., Tao Z. and Peng M. 2019. Dietary supplementation of Bacillus cereus as probiotics in Pengze crucian carp (Carassius auratus var. Pengze): Effects on growth performance, fillet quality, serum biochemical parameters and intestinal histology. Aquaculture Research, 50(8): 2207–2217.
Yu M.C., Li Z.J., Lin H.Z., Wen G.L. and Ma S. 2008. Effects of dietary Bacillus and medicinal herbs on the growth, digestive enzyme activity, and serum biochemical parameters of the shrimp Litopenaeus vannamei. Aquaculture International, 16(5): 471–480.
Zarei S., Badzohreh G., Davoodi R., Nafisi Bahabadi M. and Salehi F. 2021. Effects of dietary butyric acid glycerides on growth performance, haemato‐immunological and antioxidant status of yellowfin seabream (Acanthopagrus latus) fingerlings. Aquaculture Research, 52(11): 5840–5848.
Zhang J., Zhong L., Chi S., Chu W., Liu Y. and Hu Y. 2020. Sodium butyrate supplementation in high-soybean meal diets for juvenile rice field eel (Monopterus albus): Effects on growth, immune response and intestinal health. Aquaculture, 520: 1–9 (734952).
Zhou X., Tian Z., Wang Y. and Li W. 2010. Effect of treatment with probiotics as water additives on tilapia (Oreochromis niloticus) growth performance and immune response. Fish Physiology and Biochemistry, 36(3): 501–509.
Zhu Y., Qiu X., Ding Q., Duan M. and Wang C. 2014. Combined effects of dietary phytase and organic acid on growth and phosphorus utilization of juvenile yellow catfish Pelteobagrus fulvidraco. Aquaculture, 430: 1–8.
Zokaeifar H., Luis Balcazar J., Kamarudin M.S., Sijam K., Arshad A. and Saad C.R. 2012. Selection and identification of non-pathogenic bacteria isolated from fermented pickles with antagonistic properties against two shrimp pathogens. The Journal of Antibiotics, 65: 289–294.