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Inhibitory activity against the fish pathogen Lactococcus garvieae produced by Lactococcus lactis TW34, a lactic acid bacterium isolated from the intestinal tract of a Patagonian fish

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Abstract

After enrichment of Odontesthes platensis intestinal contents, 53 lactic acid bacteria (LAB) were isolated. From the four isolates that showed inhibitory activity against Lactococcus garvieae 03/8460, strain TW34 was selected because it exerted the strongest inhibition. It also inhibited other Gram-positive bacteria, but not Gram-negative fish pathogens. Phenotypic and 16S rDNA phylogenetic analyses showed that TW34 belongs to Lactococcus lactis. In addition, TW34 showed to be sensitive to different antibiotics. The production of the inhibitory agent against L. garvieae was growth associated, and it was significantly influenced by the incubation temperature. The optimal temperature for the antimicrobial production was as low as 15°C. Both acidification and hydrogen peroxide production were ruled out as the source of inhibition. In contrast, the antimicrobial activity was completely lost by treatment with proteolytic enzymes, which confirmed that the inhibitory substance was a bacteriocin. The bacteriocin was highly thermostable (121°C for 15 min) and active between pH 3 and 11. It remained stable for up to 2 months when stored at 4°C and up to 6 months at −20°C. Our results suggest that the strain L. lactis TW34 could provide an alternative for lactococcosis control and therefore be considered for future challenge experiments with fish.

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References

  • Altschul SF, Gish W, Miller M, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    CAS  PubMed  Google Scholar 

  • Austin B, Stuckey LF, Robertson PAW, Effendi I, Griffith DRW (1995) A probiotic strain of Vibrio alginolyticus effective in reducing diseases caused by Aeromonas salmonicida, Vibrio anguillarum and Vibrio ordalii. J Fish Dis 18:93–96

    Article  Google Scholar 

  • Balcázar JL, de Blas I, Ruiz-Zarzuela I, Cunningham D, Vendrell D, Múzquiz JL (2006) The role of probiotics in aquaculture. Vet Microbiol 114:173–186

    Article  PubMed  Google Scholar 

  • Balcázar JL, Vendrell D, de Blas I, Ruiz-Zarzuela I, Gironés O, Múzquiz JL (2007) In vitro competitive adhesion and production of antagonistic compounds by lactic acid bacteria against fish pathogens. Vet Microbiol 122:373–380

    Article  PubMed  CAS  Google Scholar 

  • Balcázar JL, Vendrell D, de Blas I, Ruiz-Zarzuela I, Muzquiz JL, Girones O (2008) Characterization of probiotic properties of lactic acid bacteria isolated from intestinal microbiota of fish. Aquaculture 278:188–191

    Article  CAS  Google Scholar 

  • Bauer AW, Kirby WMM, Sherris JC, Turk M (1966) Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 45:493–496

    CAS  PubMed  Google Scholar 

  • Bednarska M, Bednarski M, Polechoński R (2007) Current problems of streptococcal infections in fish. Med Weter 63:783–785

    Google Scholar 

  • Brillet A, Pilet MF, Prevost H, Bouttefroy A, Leroi F (2004) Biodiversity of Listeria monocytogenes sensitivity to bacteriocin-producing Carnobacterium strains and application in sterile cold-smoked salmon. J Appl Microbiol 97:1029–1037

    Article  CAS  PubMed  Google Scholar 

  • Brunt J, Austin B (2005) Use of a probiotic to control lactococcosis and streptococcosis in rainbow trout, Oncorhynchus mykiss (Walbaum). J Fish Dis 28:693–701

    Article  CAS  PubMed  Google Scholar 

  • Byun JW, Park SC, Benno Y, Oh TK (1997) Probiotic effect of Lactobacillus sp. DS-12 in flounder (Paralichthys olivaceus). J Gen Appl Microbiol 43:305–308

    Article  CAS  PubMed  Google Scholar 

  • Calo-Mata P, Arlindo S, Boehme K, de Miguel T, Pascoal A, Barros-Velazquez J (2008) Current applications and future trends of lactic acid bacteria and their bacteriocins for the biopreservation of aquatic food products. Food Bioprocess Technol 1:43–63

    Article  Google Scholar 

  • Campos CA, Rodríguez O, Calo-Mata P, Prado M, Barros-Velázquez J (2006) Preliminary characterization of bacteriocins from Lactococcus lactis, Enterococcus faecium and Enterococcus mundtii strains isolated from turbot (Psetta maxima). Food Res Int 39:356–364

    Article  CAS  Google Scholar 

  • Carson J, Gudkovs N, Austin B (1993) Characteristics of an Enterococcus-like bacterium from Australia and South Africa, pathogenic for rainbow trout (Oncorhynchus mykiss Walbaum). J Fish Dis 6:381–388

    Article  Google Scholar 

  • Chang CI, Liu WY (2002) An evaluation of two probiotic bacterial strains, Enterococcus faecium SF68 and Bacillus toyoi, for reducing edwardsiellosis in cultured European eel, Anguilla anguilla L. J Fish Dis 25:311–315

    Article  Google Scholar 

  • Chang PH, Lin CW, Lee YC (2002) Lactococcus garvieae infection of cultured rainbow trout, Oncorhynchus mykiss, in Taiwan and associated biophysical characteristics and histopathology. Bull Eur Assoc Fish Pathol 22:319–327

    Google Scholar 

  • Chatterjee C, Paul M, Xie L, van der Donk WA (2005) Biosynthesis and mode of action of lantibiotics. Chem Rev 105:633–683

    Article  CAS  PubMed  Google Scholar 

  • Cheigh CI, Choi HJ, Park H, Kim SB, Kook MC, Kim TS, Hwang JK, Pyun YR (2002) Influence of growth conditions on the production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from kimchi. J Biotechnol 95:225–235

    Article  CAS  PubMed  Google Scholar 

  • Cho SL, Nam SW, Yoon JH, Lee JS, Sukhoom A, Kim W (2008) Lactococcus chungangensis sp. nov., a lactic acid bacterium isolated from activated sludge foam. Int J Syst Evol Microbiol 58:1844–1849

    Article  CAS  PubMed  Google Scholar 

  • De Vuyst L, Callewaert R, Crabbé K (1996) Primary metabolite kinetics of bacteriocin biosynthesis by Lactobacillus amylovorus and evidence for stimulation of bacteriocin production under unfavourable growth conditions. Microbiology 142:817–827

    Article  Google Scholar 

  • Deegan LH, Cotter PD, Hill C, Ross P (2006) Bacteriocins: biological tools for bio-preservation and shelf-life extension. Int Dairy J 16:1058–1071

    Article  CAS  Google Scholar 

  • DeLong EF (1992) Archaea in coastal marine environments. Proc Natl Acad Sci USA 89:5685–5689

    Article  CAS  PubMed  Google Scholar 

  • Dykes GA, Britz TJ, von Holy A (1994) Numerical taxonomy and identification of lactic acid bacteria from spoiled, vacuum packaged Vienna sausages. J Appl Bacteriol 76:246–252

    CAS  PubMed  Google Scholar 

  • FAO (2005–2009) Cultured Aquatic Species Information Programme. Text by Cowx IG In: FAO Fisheries and Aquaculture Department (online). Rome. Updated 15 June 2005. http://www.fao.org/fishery/culturedspecies/Oncorhynchus_mykiss/en. Accessed 18 Aug 2009

  • Gatesoupe FJ (1999) The use of probiotics in aquaculture. Aquaculture 180:147–165

    Article  Google Scholar 

  • Gatesoupe FJ (2008) Updating the importance of lactic acid bacteria in fish farming: natural occurrence and probiotic treatments. J Mol Microbiol Biotechnol 14:107–114

    Article  CAS  PubMed  Google Scholar 

  • Ghittino C, Prearo M (1992) Report of streptococcosis in rainbow trout (Oncorhynchus mykiss) in Italy: preliminary note. Boll Soc It Patol Ittica 8:4–11

    Google Scholar 

  • Gilliland SE (1969) Enzymatic determination of residual hydrogen peroxide in milk. J Dairy Sci 52:321–324

    Article  CAS  Google Scholar 

  • Gomes S, Afonso A, Gartner F (2006) Fish vaccination against infections by Streptococcal species and the particular case of Lactococcosis. RPCV 101:25–35

    Google Scholar 

  • Hagi T, Tanaka D, Iwamura Y, Hoshino T (2004) Diversity and seasonal changes in lactic acid bacteria in the intestinal tract of cultured freshwater fish. Aquaculture 234:335–346

    Article  CAS  Google Scholar 

  • Irianto A, Austin B (2002) Probiotics in aquaculture. J Fish Dis 25:633–642

    Article  Google Scholar 

  • Itoi S, Abe T, Washio S, Ikuno E, Kanomata Y, Sugita H (2008) Isolation of halotolerant Lactococcus lactis subsp. lactis from intestinal tract of coastal fish. Int J Food Microbiol 121:116–121

    Article  CAS  PubMed  Google Scholar 

  • Itoi S, Yuasa K, Washio S, Abe T, Ikuno E, Sugita H (2009) Phenotypic variation in Lactococcus lactis subsp. lactis isolates derived from intestinal tracts of marine and freshwater fish. J Appl Microbiol 107:867–874

    Article  CAS  PubMed  Google Scholar 

  • Lategan MJ, Booth W, Shimmon R, Gibson LF (2006) An inhibitory substance produced by Aeromonas media A199, an aquatic probiotic. Aquaculture 254:115–124

    Article  CAS  Google Scholar 

  • Matsusaki H, Endo N, Sonomoto K, Ishizaki A (1996) Lantibiotic nisin Z fermentative production by Lactococcus lactis IO-1; relationship between production of the lantibiotic and lactate and cell growth. Appl Microbiol Biotechnol 45:36–40

    Article  CAS  PubMed  Google Scholar 

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

    Article  Google Scholar 

  • National Committee for Clinical Laboratory Standards (2000) Performance standards for antimicrobial disk susceptibility test. Approved standard M2–A7, 7th edn. National Committee for Clinical Laboratory Standards, Wayne

    Google Scholar 

  • Noonpakdee W, Santivarangkna C, Jumriangrit P, Sonomoto K, Panyim S (2003) Isolation of nisin-producing Lactococcus lactis WNC 20 strain from nham, a traditional Thai fermented sausage. Int J Food Microbiol 81:137–145

    Article  CAS  PubMed  Google Scholar 

  • Onda T, Yanagida F, Tsuji M, Shinohara T, Yokotsuka K (2003) Production and purification of a bacteriocin peptide produced by Lactococcus sp. strain GM005, isolated from Miso-paste. Int J Food Microbiol 87:153–159

    Article  CAS  PubMed  Google Scholar 

  • Parente E, Brienza C, Moles M, Ricciardi A (1995) A comparison of methods for the measurement of bacteriocin activity. J Microbiol Methods 22:95–108

    Article  Google Scholar 

  • Ringø E, Gatesoupe FJ (1998) Lactic acid bacteria in fish: a review. Aquaculture 160:177–203

    Article  Google Scholar 

  • Ringø E, Bendiksen HR, Wesmajervi MS, Olsen RE, Jansen PA, Mikkelsen H (2000) Lactic acid bacteria associated with the digestive tract of Atlantic salmon (Salmo salar L.). J Appl Microbiol 89:317–322

    Article  PubMed  Google Scholar 

  • Ryan MP, Rea MC, Hill C, Ross P (1996) An application in cheddar cheese manufacture for a strain of Lactococcus lactis producing a novel broad-spectrum bacteriocin, Lacticin 3147. Appl Environ Microbiol 62:612–619

    CAS  PubMed  Google Scholar 

  • Salama MS, Musafija-Jeknic T, Sandine WE, Giovannoni SJ (1995) An ecological study of lactic acid bacteria: isolation of new strains of Lactococcus including Lactococcus lactis subsp. cremoris. J Dairy Sci 78:1004–1017

    CAS  Google Scholar 

  • Salminen S, von Wright A, Morelli L, Marteau P, Brassart D, de Vos WM, Fondén R, Saxelin M, Collins K, Mogensen G, Birkeland SE, Mattila-Sandholm T (1998) Demonstration of safety of probiotics—a review. Int J Food Microbiol 44:93–106

    Article  CAS  PubMed  Google Scholar 

  • Spanggaard B, Huber I, Nielsen J, Sick EB, Pipper CB, Martinussen T, Slierendrecht WJ, Gram L (2001) The probiotic potential against vibriosis of the indigenous microflora of rainbow trout. Environ Microbiol 3:755–765

    Article  CAS  PubMed  Google Scholar 

  • Sugita H, Okano R, Suzuki Y, Iwai D, Mizukami M, Akiyama N, Matsuura S (2002) Antibacterial abilities of intestinal bacteria from larval and juvenile Japanese flounder against fish pathogens. Fisheries Sci 68:1004–1011

    Article  CAS  Google Scholar 

  • Sugita H, Ohta K, Kuruma A, Sagesaka T (2007) An antibacterial effect of Lactococcus lactis isolated from the intestinal tract of the Amur catfish, Silurus asotus Linnaeus. Aquac Res 38:1002–1004

    Article  Google Scholar 

  • Swofford DL (2001) PAUP*: phylogenetic analysis using parsimony (* and other methods), version 4.0b10. Sinauer Associates, Sunderland

    Google Scholar 

  • Tahiri I, Desbiens M, Benech R, Kheadr E, Lacroix C, Thibault S, Ouellet D, Fliss I (2004) Purification, characterization and amino acid sequencing of divergicin M35: a novel class IIa bacteriocin produced by Carnobacterium divergens M35. Int J Food Microbiol 97:123–136

    Article  CAS  PubMed  Google Scholar 

  • Teuber M (1995) The genus Lactococcus. In: Wood BJB, Holzapfel WH (eds) The genera of lactic acid bacteria. Chapman & Hall, Glasgow, pp 173–234

    Google Scholar 

  • Vázquez JA, González MP, Murado MA (2005) Effects of lactic acid bacteria cultures on pathogenic microbiota from fish. Aquaculture 245:149–161

    Article  Google Scholar 

  • Vendrell D, Balcázar JL, Ruiz-Zarzuela I, de Blas I, Gironés O, Múzquiz JL (2006) Lactococcus garvieae in fish: a review. Comp Immunol Microbiol 29:177–198

    Article  Google Scholar 

  • Vendrell D, Balcázar JL, de Blas I, Ruiz-Zarzuela I, Gironés O, Múzquiz JL (2008) Protection of rainbow trout (Oncorhynchus mykiss) from lactococcosis by probiotic bacteria. Comp Immunol Microbiol 31:337–345

    Article  Google Scholar 

  • Villamil L, Figueras A, Planas M, Novoa B (2003) Control of Vibrio alginolyticus in Artemia culture by treatment with bacterial probiotics. Aquaculture 219:43–56

    Article  Google Scholar 

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Acknowledgments

The authors acknowledge Dr. Jose Francisco Fernandez-Garayzabal for kindly supplying the indicator strains. This research was supported by the Agencia de Promoción Científica y Tecnológica, Ministerio de Educación Ciencia y Tecnología, Argentina, PICT No 32897. C. Sequeiros and M. Vallejo thank FONCyT (Fondo para la Investigación Científica y Tecnológica, Argentina) and CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina), respectively, for Postdoctoral Fellowships.

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Correspondence to Cynthia Sequeiros.

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Communicated by Erko Stackebrandt.

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Sequeiros, C., Vallejo, M., Marguet, E.R. et al. Inhibitory activity against the fish pathogen Lactococcus garvieae produced by Lactococcus lactis TW34, a lactic acid bacterium isolated from the intestinal tract of a Patagonian fish. Arch Microbiol 192, 237–245 (2010). https://doi.org/10.1007/s00203-010-0552-1

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