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Probiotics in the intestinal tract of juvenile whiteleg shrimp Litopenaeus vannamei: modulation of the bacterial community

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Abstract

Molecular analysis of the 16S rDNA of the intestinal microbiota of whiteleg shrimp Litopenaeus vannamei was examined to investigate the effect of a Bacillus mix (Bacillus endophyticus YC3-b, Bacillus endophyticus C2-2, Bacillus tequilensisYC5-2) and the commercial probiotic (Alibio®) on intestinal bacterial communities and resistance to Vibrio infection. PCR and single strain conformation polymorphism (SSCP) analyses were then performed on DNA extracted directly from guts. Injection of shrimp with V. parahaemolyticus at 2.5 × 105 CFU g−1 per shrimp followed 168 h after inoculation with Bacillus mix or the Alibio probiotic or the positive control. Diversity analyses showed that the bacterial community resulting from the Bacillus mix had the highest diversity and evenness and the bacterial community of the control had the lowest diversity. The bacterial community treated with probiotics mainly consisted of α- and γ-proteobacteria, fusobacteria, sphingobacteria, and flavobacteria, while the control mainly consisted of α-proteobacteria and flavobacteria. Differences were grouped using principal component analyses of PCR-SSCP of the microbiota, according to the time of inoculation. In Vibrio parahaemolyticus-infected shrimp, the Bacillus mix (~33 %) induced a significant increase in survival compared to Alibio (~21 %) and the control (~9 %). We conclude that administration of the Bacillus mix induced modulation of the intestinal microbiota of L. vannamei and increased its resistance to V. parahaemolyticus.

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References

  • Amann RI, Ludwig W, Schleifer KH (1995) Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev 59:143–169

    CAS  Google Scholar 

  • Bachére E (2000) Introduction: shrimp immunity and disease control. Aquaculture 191:3–11

    Article  Google Scholar 

  • Benbouza H, Jacquemin JM, Baudoin JP, Mergeai G (2006) Optimization of a reliable, fast, cheap and sensitive silver staining method to detect SSR markers in polyacrylamide gels. Biotechnol Agron Soc 10:77–81

    CAS  Google Scholar 

  • Chaiyapechara S, Rungrassamee W, Suriyachay I (2011) Bacterial community associated with the intestinal tract of P. monodon in commercial farms. Microb Ecol. doi:10.1007/s00248-011-9936-2

    Google Scholar 

  • Chun J, Lee JH, Jung Y, Kim M, Kim S, Kim BK, Lim YW (2007) EzTaxon: a web-based tool for the identification of prokaryotes based on 16S ribosomal RNA gene sequences. Int J Syst Evol Micr 57:2259–2261

    Article  CAS  Google Scholar 

  • Chythanya R, Karunasagar I, Karunasagar I (2002) Inhibition of shrimp pathogenic vibrios by a marine Pseudomonas I-2 strain. Aquaculture 208:1–10

    Article  Google Scholar 

  • Daniels CL, Merrifield DL, Boothroyd DP, Davies SJ, Factor JR, Arnold KE (2010) Effect of dietary Bacillus spp. and mannan oligosaccharides (MOS) on European lobster (Homarus gammarus L.) larvae growth performance, gut morphology and gut microbiology and gut microbiota. Aquaculture 304:49–57

    Article  CAS  Google Scholar 

  • Dempsey AC, Kitting CI, Rosson RA (1989) Bacterial variability individual panaeid shrimp digestive tracts. Crustaceana 56:267–278

    Article  Google Scholar 

  • Dethlefsen L, Huse S, Sogin ML, Realman DA (2008) The pervasive effects of an antibiotic on the human gut microbiota, as revealed by deep 16S rRNA sequencing. PLoS Biol 6:2383–2400

    Article  CAS  Google Scholar 

  • Dohrmann AB, Tebbe CC (2004) Microbial community analysis by PCR single strand conformation polymorphism (PCR-SSCP), PCR single strand conformation polymorphism (PCR-SSCP). In: Kowalchuk GA, Bruijn FJ, Head IM, Akkermans AD, van Elsas JD (eds) Molecular microbial ecology manual. Kluwer Academic Publishers, Dordrecht, pp 809–838

    Google Scholar 

  • Drablos F, Nochilson D, Ronning M (1999) EXAFS study of zinc coordination in Bacitracin A. Biochim Biophys Acta 1431:433–442

    Article  CAS  Google Scholar 

  • Durand L, Zbinden M, Cueff − Gauchard V, Duperron S, Roussell EG, Shillito B, Cambon − Bonavita MA (2010) Microbial diversity associated with the hydrothermal shrimp Rimicaris exoculata gut and occurrence of a resident microbial community. FEMS Microbiol Ecol 71:291–303

    Article  CAS  Google Scholar 

  • Escobar-Briones L, Olvera-Novoa MA, Puerto-Castillo C (2006) Avances sobre ecología microbiana del tracto digestivo de la tilapia y sus potenciales implicaciones. VIII Simposium internacional de Nutrición Acuícola. Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, Mexico

  • Farzanfar A (2006) The use of probiotics in shrimp aquaculture. FEMS Immunol Med Microbiol 48:149–158

    Article  CAS  Google Scholar 

  • Fromin N, Hamelin J, Tarnawski S, Roesti D, Jourdain-Miserez K, Forestier N, Teyssier-Cuvelle S, Gillet F, Aragno M, Rossi P (2002) Statistical analysis of denaturing gel electrophoresis (DGE) fingerprinting patterns. Environ Microbiol 4:634–643

    Google Scholar 

  • Gelsomino A, Keijzer WAC, Cacco G, Van Elsas JD (1999) Assessment of bacterial community structure in soil by polymerase chain reaction and denaturing gradient gel electrophoresis. J Microbiol Methods 38:1–15

    Article  CAS  Google Scholar 

  • Guo P, Xu M (1994) The bacterial variation in the water environment of cultured prawn pond. Oceano et Limno Sinica 25:625–629

    Google Scholar 

  • Holzapfel W, Haberer P, Snel J, Schillinger U (1988) Overview of gut flora and probiotics. Int J Food Microbiol 41:85–101

    Article  Google Scholar 

  • Johnson CN, Barnes S, Ogle J, Grimes JD (2008) Microbial community analysis of water, foregut, and hindgut during growth of Pacific white shrimp, Litopenaeus vannamei, in closed-system aquaculture. J World Aquacult Soc 39:251–257

    Article  Google Scholar 

  • Lau WWY, Jumars PA, Armbrust EV (2002) Genetic diversity of attached bacteria in the hindgut of the deposit-feeding shrimp Neotrypaea (formerly Callianassa) californiensis (Decapoda: Thalassinidae). Microb Ecol 43:455–466

    Article  CAS  Google Scholar 

  • Lightner DV (2005) Biosecurity in shrimp farming: pathogen exclusion through use of SPF stock and routine surveillance. J World Aquacult Soc 36:229–248

    Article  Google Scholar 

  • Liu CH, Chiu CS, Ho PL, Wang SW (2009) Improvement in the growth performance of white shrimp, Litopenaeus vannamei, by a protease-producing probiotic, Bacillus subtilis E20, fronnatto. J Appl Microbiol 107:1031–1041

    Article  CAS  Google Scholar 

  • Liu H, Liu M, Wang B, Jiang K, Jiang S, Sun S, Wang L (2010) PCR-DGGE analysis of intestinal bacteria and effect of Bacillus spp. On intestinal microbial diversity in kuruma shrimp (Marsupenaeus japonicus). Chin J Oceanol Limn 28:808–814

    Article  CAS  Google Scholar 

  • Liu H, Wang L, Liu M, Wang B, Jiang K, Ma S, Li Q (2011) The intestinal microbial diversity in Chinese shrimp (Fenneropenaeus chinensis) as determined by PCR-DGGE and clone library analyses. Aquaculture 317:32–36

    Article  CAS  Google Scholar 

  • Luis-Villaseñor IE, Macias-Rodriguez ME, Gomez-Gil B, Ascencio F, Campa-Cordova AI (2011) Beneficial effects of four Bacillus strains on the larval cultivation of Litopenaeus vannamei. Aquaculture 321:136–144

    Article  Google Scholar 

  • Luo P, Hu C, Xie Z, Zhang L, Ren C, Xu Y (2006) PCR-DGGE analysis of bacterial community composition in brackish water Litopenaeus vannamei culture system. J Trop Oceanogr 25:49–53

    CAS  Google Scholar 

  • Moriarty DJW (1998) Control of luminous Vibrio species in penaeid aquaculture ponds. Aquaculture 164:351–358

    Article  Google Scholar 

  • Morikawa M, Ito M, Imanaka T (1992) Isolation of a new surfactin producer Bacillus pumilus A-1, and cloning and nucleotide sequence of the regulator gene, psf-1. J Ferment Bioeng 74:255–261

    Article  CAS  Google Scholar 

  • Payne MS, Hall MR, Sly L, Bourne DG (2007) Microbial diversity within early-stage cultured Panulirus ornatusphyllosomas. Appl Environ Microbiol 73:1940–1951

    Article  CAS  Google Scholar 

  • Perez C, Suarez C, Castro GR (1993) Antimicrobial activity determined in strains of Bacillus circulans cluster. Folia Microbiol 38:25–28

    Article  CAS  Google Scholar 

  • Rodriguez J, Espinosa Y, Echeverria F, Cardenas G, Roman R, Stern S (2007) Exposure to probiotics and b-1,3 / 1,6-glucans in larviculture modifies the immune response of Penaeus vannamei juveniles and both the survival to White Spot Syndrome Virus challenge and pond culture. Aquaculture 273:405–415

    Article  CAS  Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Schwiger F, Tebbe CC (1998) A new approach to utilize PCR-single strand-conformation polymorphism for 16S rRNA gene-based microbial community analysis. Appl Environ Microbiol 64:4870–4876

    Google Scholar 

  • Tapia-Paniagua ST, Chabrillón M, Díaz,-Rosales P, de la Banda IG, Lobo C, Balebona MC, Moringo MA (2010) Intestinal microbiota diversity of the flat fish Solea senegalensis (Kaup, 1858) following probiotic administration. Microb Ecol 60:310–319

    Google Scholar 

  • Ziaei NS, Rezaei MH, Takami GA, Lovett DL, Mirvaghefi AR, Shakouri M (2006) The effect of Bacillus spp. bacteria used as probiotics on digestive enzyme activity, survival and growth in the Indian white shrimp Fenneropenaeus indicus. Aquaculture 252:516–524

    Article  Google Scholar 

Download references

Acknowledgments

The authors thank Maria de Jésus Romero Geraldo of CIBNOR for assistance. Ira Fogel of CIBNOR provided editorial services. The project was funded by Consejo Nacional de Ciencia y Tecnología, SEP-CONACYT grant 25981, and Centro de Investigaciones Biológicas del Noroeste grant AC0.4. I.E.L.V. is a recipient of a CONACYT fellowship (grant 48351).

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Correspondence to Felipe Ascencio.

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Luis-Villaseñor, I.E., Castellanos-Cervantes, T., Gomez-Gil, B. et al. Probiotics in the intestinal tract of juvenile whiteleg shrimp Litopenaeus vannamei: modulation of the bacterial community. World J Microbiol Biotechnol 29, 257–265 (2013). https://doi.org/10.1007/s11274-012-1177-0

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  • DOI: https://doi.org/10.1007/s11274-012-1177-0

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