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Improved antimicrobial activity of Pediococcus acidilactici against Salmonella Gallinarum by UV mutagenesis and genome shuffling

  • Applied genetics and molecular biotechnology
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Abstract

Pediococcus acidilactici is a widely used probiotic, and Salmonella enterica serovar Gallinarum (SG) is a significant pathogen in the poultry industry. In this study, we improved the antimicrobial activity of P. acidilactici against SG using UV mutation and genome shuffling (GS). To improve antimicrobial activity against SG, UV mutagenesis was performed against wild-type P. acidilactici (WT), and five mutants showed improved antimicrobial activity. To further improve antimicrobial activity, GS was performed on five UV mutants. Following GS, four mutants showed improved antimicrobial activity compared with the UV mutants and WT. The antimicrobial activity of GS1 was highest among the mutants; however, the activity was reduced when the culture supernatant was treated with proteinase K, suggesting that the improved antimicrobial activity is due to a proteinous substance such as bacteriocin. To validate the activity of GS1 in vivo, we designed multi-species probiotics and performed broiler feeding experiments. Groups consisted of no treatment (NC), avilamycin-treated (PC), probiotic group 1 containing WT (T1), and probiotic group 2 containing GS1 (T2). In broiler feeding experiments, coliform bacteria were significantly reduced in T2 compared with NC, PC, and T1. The cecal microbiota was modulated and pathogenic bacteria were reduced by GS1 oral administration. In this study, GS1 showed improved antimicrobial activity against SG in vitro and reduced pathogenic bacteria in a broiler feeding experiment. These results suggest that GS1 can serve as an efficient probiotic, as an alternative to antibiotics in the poultry industry.

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References

  • Adam JK, Bharti O, Naidu KSB (2012) Probiotics: recent understandings and biomedical applications. Curr Trends Biotechnol Pharma 6:1–14

    Google Scholar 

  • Andersson A, Ronner U, Granum PE (1995) What problems does the food industry have with the spore-forming pathogens Bacillus cereus and Clostridium perfringens? Int J Food Microbiol 28:145–155

    Article  CAS  PubMed  Google Scholar 

  • Bogaert D, de Groot R, Hermans PWM (2004) Streptococcus pneumoniae colonisation: the key to pneumococcal disease. Lancet Infect Dis 4:144–154

    Article  CAS  PubMed  Google Scholar 

  • Chanos P, Mygind T (2016) Co-culture-inducible bacteriocin production in lactic acid bacteria. Appl Microbiol Biotechnol 100:4297–4308

    Article  CAS  PubMed  Google Scholar 

  • Cocconcelli PS, Morelli L, Vescovo M, Bottazzi V (1986) Intergeneric protoplast fusion in lactic-acid bacteria. FEMS Microbiol Lett 35:211–214

    Article  CAS  Google Scholar 

  • Cornillot E, Saintjoanis B, Daube G, Katayama S, Granum PE, Canard B, Cole ST (1995) The enterotoxin gene (cpe) of Clostridium perfringens can be chromosomal or plasmid-borne. Mol Microbiol 15:639–647

    Article  CAS  PubMed  Google Scholar 

  • Dai MH, Ziesman S, Ratcliffe T, Gill RT, Copley SD (2005) Visualization of protoplast fusion and quantitation of recombination in fused protoplasts of auxotrophic strains of Escherichia coli. Metab Eng 7:45–52

    Article  CAS  PubMed  Google Scholar 

  • Famularo G, De Simone C, Matteuzi D, Pirovano F (1999) Traditional and high potency probiotic preparations for oral bacteriotherapy. BioDrugs 12:455–470

    Article  CAS  PubMed  Google Scholar 

  • Fuller R (1989) Probiotics in man and animals. J Appl Bacteriol 66:365–378

    Article  CAS  PubMed  Google Scholar 

  • Gallagher T, Spino D (1968) The significance of numbers of coliform bacteria as an indicator of enteric pathogens. Water Res 2:169–175

    Article  Google Scholar 

  • Granum PE (1990) Clostridium perfringens toxins involved in food poisoning. Int J Food Microbiol 10:101–112

    Article  CAS  PubMed  Google Scholar 

  • Hegazy AM, Barakat M, Fadl SE, El-Keredy AM (2014) Effect of Pedicoccus acidilactici on immunity, production and lipid profile in broilers. Alexandria J Vet Sci 41:35–46

    Article  Google Scholar 

  • Heng NC, Wescombe PA, Burton JP, Jack RW, Tagg JR (2007) The diversity of bacteriocins in Gram-positive bacteria. In: Riley MA, Chavan MA (eds) Bacteriocins. Springer, Berlin, pp 45–92

    Chapter  Google Scholar 

  • Johnson M, Hanlin M, Ray B (1992) Low pH and lactate are necessary for conversion of prepediocin to pediocin AcH in Pediococcus acidilactici H. In: Annu Meeting 1992, New Orleans, LA, May 26–30. Abstr, 1992

  • Kelesidis T, Pothoulakis C (2012) Efficacy and safety of the probiotics Saccharomyces boulardii for the prevetion and therapy of gastrointestinal disorders. Ther Adv Gastroenterol 5:111–125

    Article  Google Scholar 

  • Lee S, Lillehoj HS, Park DW, Hong YH, Lin J (2007) Effects of Pediococcus- and Saccharomyces-based probiotic (MitoMax) on coccidiosis in broiler chickens. Comp Immunol Microbiol Infect Dis 30:261–268

    Article  PubMed  Google Scholar 

  • Matsuda K, Chaudhari AA, Lee JH (2011) Evaluation of safety and protection efficacy on cpxR and lon deleted mutant of Salmonella Gallinarum as a live vaccine candidate for fowl typhoid. Vaccine 29:668–674

    Article  CAS  PubMed  Google Scholar 

  • Moore JE, Corcoran D, Dooley JSG, Fanning S, Lucey B, Matsuda M, McDowell DA, Mégraud F, Millar BC, O’Mahony R, O’Riordan L, O’Rourke M, Rao JR, Rooney PJ, Sails A, Whyte P (2005) Campylobacter. Vet Res 36:351–382

    Article  CAS  PubMed  Google Scholar 

  • Morelli L, Cocconcelli PS, Bottazzi V, Damiani G, Ferretti L, Sgaramella V (1987) Lactobacillus protoplast transformation. Plasmid 17:73–75

    Article  CAS  PubMed  Google Scholar 

  • Nahashon SN, Nakaue HS, Mirosh LW (1994) Production variables and nutrient retention in single comb white leghorn laying pullets fed diets supplemented with direct-fed microbials. Poult Sci 73:1699–1711

    Article  CAS  PubMed  Google Scholar 

  • Patnaik R, Louie S, Gavrilovic V, Perry K, Stemmer WPC, Ryan CM, del Cardayre S (2002) Genome shuffling of Lactobacillus for improved acid tolerance. Nat Biotechnol 20:707–712

    Article  CAS  PubMed  Google Scholar 

  • Pucci MJ, Vedamuthu ER, Kunka BS, Vandenbergh PA (1988) Inhibition of Listeria monocytogenes by using bacteriocin PA-1 produced by Pediococcus acidilactici PAC 1.0. Appl Environ Microbiol 54:2349–2353

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ray B (1995) Pediococcus in fermented foods. In: Hui YH, Khachatourlans GG (eds) Food biotechnology: microorganisms. VCH Publishers, New York, pp 745–795

    Google Scholar 

  • Shi DJ, Wang CL, Wang KM (2009) Genome shuffling to improve thermotolerance, ethanol tolerance and ethanol productivity of Saccharomyces cerevisiae. J Ind Microbiol Biotechnol 36:139–147

    Article  CAS  PubMed  Google Scholar 

  • Timmerman HM, Koning CJM, Mulder L, Rombouts FM, Beynen AC (2004) Monostrain, multistrain and multispecies problotics—a comparison of functionality and efficacy. Int J Food Microbiol 96:219–233

    Article  CAS  PubMed  Google Scholar 

  • Wang HK, Zhang J, Wang XJ, Qi W, Dai YJ (2012) Genome shuffling improves production of the low-temperature alkalophilic lipase by Acinetobacter johnsonii. Biotechnol Lett 34:145–151

    Article  PubMed  Google Scholar 

  • Yu L, Pei X, Lei T, Wang Y, Feng Y (2008) Genome shuffling enhanced L-lactic acid production by improving glucose tolerance of Lactobacillus rhamnosus. J Biotechnol 134:154–159

    Article  CAS  PubMed  Google Scholar 

  • Zhang-Barber L, Turner AK, Dougan G, Barrow PA (1998) Protection of chickens against experimental fowl typhoid using a nuoG mutant of Salmonella serotype Gallinarum. Vaccine 16:899–903

    Article  CAS  PubMed  Google Scholar 

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Author notes

  1. Geon Goo Han and Ahn Ah Song contributed equally to this work.

Authors and Affiliations

  1. Department of Agricultural Biotechnology, Seoul National University, Seoul, Republic of Korea

    Geon Goo Han, Ahn Ah Song, Seong-Hyun Yoon, Chong-Su Cho & Yun-Jaie Choi

  2. Department of Animal Life Science, Kangwon National University, Chuncheon, Gangwon-do, Republic of Korea

    Eun Bae Kim

  3. Institute of Green-Bio Science & Technology, Seoul National University, Pyeongchang, Gangwon-do, Republic of Korea

    **-Duck Bok & Sang-Kee Kang

  4. Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Republic of Korea

    Chong-Su Cho & Yun-Jaie Choi

  5. Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do, Republic of Korea

    Dong Yong Kil

Authors
  1. Geon Goo Han
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  2. Ahn Ah Song
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  3. Eun Bae Kim
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  5. **-Duck Bok
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  8. Sang-Kee Kang
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  9. Yun-Jaie Choi
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Corresponding authors

Correspondence to Sang-Kee Kang or Yun-Jaie Choi.

Ethics declarations

The Institutional Animal Care and Use Committee at Chung-Ang University approved animal experiments under protocol #14-0005.

Funding

This study was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through Agri-Bio industry Technology Development Program, funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA) (115084-2 [Yun-Jaie Choi] and 316005-5 [Sang-Kee Kang]). Geon Goo Han was supported by the BK21 Plus program.

Conflict of interest

The authors declare that they have no conflict of interests.

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Han, G.G., Song, A.A., Kim, E.B. et al. Improved antimicrobial activity of Pediococcus acidilactici against Salmonella Gallinarum by UV mutagenesis and genome shuffling. Appl Microbiol Biotechnol 101, 5353–5363 (2017). https://doi.org/10.1007/s00253-017-8293-6

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  • DOI: https://doi.org/10.1007/s00253-017-8293-6

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