Skip to main content
SpringerLink
Log in

Selection of Potential Probiotic Lactobacillus with Inhibitory Activity Against Salmonella and Fecal Coliform Bacteria

  • Published:
Probiotics and Antimicrobial Proteins Aims and scope Submit manuscript

Abstract

Three hundred and sixty presumptive lactic acid bacteria (LAB) isolated from pregnant sows, newborn, suckling, and weaned piglets were preliminarily screened for anti-Salmonella activity. Fifty-eight isolates consisting of Lactobacillus reuteri (n = 32), Lactobacillus salivarius (n = 10), Lactobacillus mucosae (n = 8), Lactobacillus johnsonii (n = 5), and Lactobacillus crispatus (n = 3) were selected and further characterized for probiotic properties including production of antimicrobial substances, acid and bile tolerance, and cell adherence to Caco-2 cells. Eight isolates including Lact. johnsonii LJ202 and Lact. reuteri LR108 were identified as potential probiotics. LJ202 was selected for further use in co-culture studies of two-bacterial and multiple-bacterial species to examine its inhibitory activity against Salmonella enterica serovar Enteritidis DMST7106 (SE7106). Co-culture of LJ202 and SE7106 showed that LJ202 could completely inhibit the growth of SE7106 in 10 h of co-culture. In co-culture of multiple-bacterial species, culturable fecal bacteria from pig feces were used as representative of multiple-bacterial species. The study was performed to examine whether interactions among multiple-bacterial species would influence antagonistic activity of LJ202 against SE7106 and fecal coliform bacteria. Co-culture of SE7106 with different combinations of fecal bacteria and probiotic (LJ202 and LR108) or non-probiotic (Lact. mucosae LM303) strains revealed that the growth of SE7106 was completely inhibited either in the presence or in the absence of probiotic strains. Intriguingly, LJ202 exhibited notable inhibitory activity against fecal coliform bacteria while LR108 did not. Taken together, the results of co-culture studies suggested that LJ202 is a good probiotic candidate for further study its inhibitory effects against pathogen infections in pigs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Kreuzer S, Janczyk P, Assmus J, Schmidt MF, Brockmann GA, Nockler K (2012) No beneficial effects evident for Enterococcus faecium NCIMB 10415 in weaned pigs infected with Salmonella enterica serovar Typhimurium DT104. Appl Environ Microbiol 78:4816–4825

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Mathew AG, Cissell R, Liamthong S (2007) Antibiotic resistance in bacteria associated with food animals: a United States perspective of livestock production. Foodborne Pathog Dis 4:115–133

    Article  CAS  PubMed  Google Scholar 

  3. Regulation 1831/2003/EC. Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 September 2003 on additives for use in animal nutrition. Official Journal of the European Union 268

  4. FAO/WHO (2001) Health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. FAO/WHO, Rome

    Google Scholar 

  5. Veizaj-Delia E, Piu T, Lekaj P, Tafaj M (2010) Using combined probiotic to improve growth performance of weaned piglets on extensive farm conditions. Livest Sci 134:249–251

    Article  Google Scholar 

  6. Wang J, Ji HF, Hou CL, Wang SX, Zhang DY, Liu H, Shan DC, Wang YM (2014) Effects of Lactobacillus johnsonii X54 supplementation on reproductive performance, gut environment, and blood biochemical and immunological index in lactating sows. Livest Sci 164:96–101

    Article  Google Scholar 

  7. Lähteinen T, Lindholm A, Rinttilä T, Junnikkala S, Kant R, Pietilä TE, Levonen K, von Ossowski I, Solano-Aguilar G, Jakava-Viljanen M, Palva A (2014) Effect of Lactobacillus brevis ATCC 8287 as a feeding supplement on the performance and immune function of piglets. Vet Immunol Immunopathol 158:14–25

    Article  CAS  PubMed  Google Scholar 

  8. Lähteinen T, Rinttilä T, Koort JMK, Kant R, Levonen K, Jakava-Viljanen M, Björkroth J, Palva A (2015) Effects of a multispecies Lactobacillus formation as a feeding supplement on the performance and immune function of piglets. Livest Sci 180:164–171

    Article  Google Scholar 

  9. Higgins JP, Higgins SE, Wolfenden AD, Henderson SN, Torres-Rodiguez A, Vicente JL, Hargis BM, Tellez G (2010) Effect of lactic acid bacteria probiotic culture treatment timing on Salmonella Enteritidis in neonatal broilers. Poult Sci 89:243–247

    Article  CAS  PubMed  Google Scholar 

  10. Chen CY, Tsen HY, Lin CL, Yu B, Chen CS (2012) Oral administration of a combination of select lactic acid bacteria strains to reduce the Salmonella invasion and inflammation of broiler chicks. Poult Sci 91:2139–2147

    Article  CAS  PubMed  Google Scholar 

  11. Feng J, Wang L, Zhou L, Yang X, Zhao X (2016) Using in vitro immunomodulatory properties of lactic acid bacteria for selection of probiotics against Salmonella infection in broiler chicks. PLoS One 11(1):e0147630. doi:10.1371/journal. pone.0147630

    Article  PubMed  PubMed Central  Google Scholar 

  12. Tsai CC, Hsih HY, Chiu HH, Lai YY, Liu JH, Yu B, Tsen HY (2005) Antagonistic activity against Salmonella infection in vitro and in vivo for two Lactobacillus strains from swine and poultry. Int J Food Microbiol 102:185–194

    Article  PubMed  Google Scholar 

  13. Lin CK, Tsai HC, Lin PP, Tsen HY, Tsai CC (2008) Lactobacillus acidophilus LAP5 able to inhibit the Salmonella Choleraesuis invasion to the human Caco-2 epithelial cell. Anaerobe 14:251–255

    Article  CAS  PubMed  Google Scholar 

  14. Casey PG, Gardiner GE, Casey G, Bradshaw B, Lawlor PG, Lynch PB, Leonard FC, Stanton C, Ross RP, Fitzgerald GF, Hill C (2007) A five-strain probiotic combination reduced pathogen shedding and alleviates disease signs in pigs challenged with Salmonella enterica serovar Typhimurium. Appl Environ Microbiol 73:1858–1863

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Barbosa TM, Serra CR, La Ragione RM, Woodward MJ, Henriques AO (2005) Screening for Bacillus isolates in the broiler gastrointestinal tract. Appl Environ Microbiol 71:968–978

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Schillinger U, Lücke FK (1989) Antibacterial activity of Lactobacillus sake isolated from meat. Appl Environ Microbiol 55:1901–1906

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Chauvière G, Coconnier MH, Kernéis S, Fourniat J, Servin AL (1992) Adhesion of human Lactobacillus acidophilus strain LB to human enterocyte-like Caco-2 cells. J Gen Microbiol 138:1689–1696

    Article  PubMed  Google Scholar 

  18. WigneswaranV, AmadorCI, JelsbakL, SternbergC, JelsbakL (2016) Utilization and control of ecological interactions in polymicrobial infections and community-based microbial cell factories [version 1; referees:3 approved]. F1000 Research. doi:10.12688/f1000research.7878.1

  19. Leser TD, Amenuvor JZ, Jensen TK, Lindecrona RH, Boye M, Møller K (2002) Culture-independent analysis of gut bacteria: the pig gastrointestinal tract microbiota revisited. Appl Environ Microbiol 68:673–690

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Yang F, Wang A, Zeng X, Hou C, Liu H, Qiao S (2015) Lactobacillus reuteri I5007 modulates tight junction protein expression in IPEC-J2 cells with LPS stimulation and in newborn piglets under normal conditions. BMC Microbiol 15:32. doi:10.1186/s12866-015-0372-1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Zhang J, Deng J, Wang Z, Che C, Li YF, Yang Q (2011) Modulatory effects of Lactobacillus salivarius on intestinal mucosal immunity of piglets. Curr Microbiol 62:1623–1631

    Article  CAS  PubMed  Google Scholar 

  22. Rubin HE, Nerad T, Vaughan F (1982) Lactate acid inhibition of Salmonella Typhimurium in yogurt. J Dairy Sci 65:197–203

    Article  CAS  PubMed  Google Scholar 

  23. Fayol-Messaoudi D, Berger CN, Coconnier-Polter MH, Liévin-LeMoal V, Servin AL (2005) pH-, lactic acid-, and non-lactic acid-dependent activities of probiotic Lactobacillus against Salmonella enterica serovar Typhimurium. Appl Environ Microbiol 71:6008–6013

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Nouaille S, Even S, Charlier C, Le Loir Y, Cocaign-Bousquet M, Loubière P (2009) Transcriptome response of Lactococcus lactis in mixed culture with Staphylococcus aureus. Appl Environ Microbiol 75:4473–4482

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Savage DC (1977) Microbiology of the gastrointestinal tract. Annu Rev Microbiol 31:107–133

    Article  CAS  PubMed  Google Scholar 

  26. De Vuyst L, Leroy F (2007) Bacteriocins from lactic acid bacteria: production, purification, and food applications. J Mol Microbiol Biotechnol 13:194–199

    Article  CAS  PubMed  Google Scholar 

  27. Melin L, Mattsson S, Katouli M, Wallgren P (2004) Development of post-weaning diarrhoea in piglets. Relation to presence of Escherichia coli strains and rotavirus. J Vet Med B Infect Dis Vet Public Health 51:12–22

    Article  CAS  PubMed  Google Scholar 

  28. Shu Q, Qu F, Gill HS (2001) Probiotic treatment using Bifidobacterium lactis HN019 reduces weanling diarrhoea associated with rotavirus and Escherichia coli infection in a piglet model. J Pediatr Gastroenterol Nutr 33:171–177

    Article  CAS  PubMed  Google Scholar 

  29. Huang CH, Qiao SY, Li DF, Piao XS, Ren JP (2004) Effects of lactobacilli on the performance, diarrhea incidence, VFA concentration and gastrointestinal microbial flora of weaning pigs. Asian Australas J Anim Sci 17:401–409

    Article  Google Scholar 

  30. Mallo JJ, Rioperez J, Honrubia P (2010) The addition of Enterococcus faecium to diets improves piglet’s intestinal microbiota and performance. Livest Sci 133:176–178

    Article  Google Scholar 

  31. Liu H, Ji HF, Zhang DY, Wang SX, Wang J, Shan DC, Wang YM (2015) Effects of Lactobacillus brevis preparation on growth performance, fecal microflora and serum profile in weaned pigs. Livest Sci 178:251–254

    Article  Google Scholar 

  32. Peng Q, Zeng XF, Zhu JL, Wang S, Liu XT, Hou CL, Thacker PA, Qiao SY (2016) Effects of dietary Lactobacillus plantarum B1 on growth performance, intestinal microbiota, and short chain fatty acid profiles in broiler chickens. Poult Sci 95:893–900

    Article  CAS  PubMed  Google Scholar 

  33. Wang S, Peng Q, Jia HM, Zeng XF, Zhu JL, Hou CL, Liu XT, Yang FJ, Qiao SY (2017) Prevention of Escherichia coliinfection in broiler chickens with Lactobacillus plantarum B1. Poult Sci. doi:10.3382/ps/pex061

  34. Haberbeck LU, Oliveira RC, Vivijis B, Wenseleers T, Aertsen A, Michiels C, Geeraerd AH (2015) Viability in growth/no growth boundaries of 188 different Escherichia colistrains reveals that approximately 75% have a higher growth probability under low pH conditions than E. coliO157:H7 strain ATCC 43888. Food Microbiol 45:222–230

    Article  CAS  PubMed  Google Scholar 

  35. Abee T, Klaenhammer TR, Letellier L (1994) Kinetic studies of the action of lactacin F, a bacteriocin produced by Lactobacillus johnsonii that forms poration complexs in the cytoplasmic membrane. Appl Environ Microbiol 60:1006–1013

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Drissi F, Merhej V, Blanc-Tailleur C, Raoult D (2015) Draft genome sequence of the Lactobacillus mucosae Marseille. Genome Announc 3(4):e00841–e00815. doi:10.1128/genomeA.00841-15

    Article  PubMed  PubMed Central  Google Scholar 

  37. Ramsey MM, Rumbaugh KP, Whiteley M (2011) Metabolite cross-feeding enhances virulence in a model polymicrobial infection. PLoS Pathog 7(3):e1002012. doi:10.1371/journaLact.ppat.1002012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Chubiz LM, Granger BR, Segrè D, Harcombe WR (2015) Species interactions differ in their genetic robustness. Front Microbiol 6:271. doi:10.3389/fmicb.2015.00271

    Article  PubMed  PubMed Central  Google Scholar 

  39. Chanter N (1997) Streptococci and enterococci as animal pathogens. J Appl Microbiol Symp Suppl 83:100S–109S

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was funded by the National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand (grant number P-14-50177). We thank Ms. Sukitaya Veeranondha for her assistance in the culture of Caco-2 cells.

Author information

Authors and Affiliations

  1. Food Biotechnology Laboratory, Food Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani, 12120, Thailand

    Mattika Abhisingha, Jureeporn Dumnil & Chetsadaporn Pitaksutheepong

Authors
  1. Mattika Abhisingha
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jureeporn Dumnil
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chetsadaporn Pitaksutheepong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chetsadaporn Pitaksutheepong.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Abhisingha, M., Dumnil, J. & Pitaksutheepong, C. Selection of Potential Probiotic Lactobacillus with Inhibitory Activity Against Salmonella and Fecal Coliform Bacteria. Probiotics & Antimicro. Prot. 10, 218–227 (2018). https://doi.org/10.1007/s12602-017-9304-8

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12602-017-9304-8

Keywords

Search

Navigation