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Probiotics and Antimicrobial Proteins

, Volume 10, Issue 2, pp 383–390 | Cite as

Protective Effects of Cell-Free Supernatant and Live Lactic Acid Bacteria Isolated from Thai Pigs Against a Pandemic Strain of Porcine Epidemic Diarrhea Virus

  • Wandee Sirichokchatchawan
  • Gun Temeeyasen
  • Dachrit Nilubol
  • Nuvee Prapasarakul
Article

Abstract

Porcine epidemic diarrhea virus (PEDV) is a coronavirus which causes severe diarrhea and fatal dehydration in piglets. In general, probiotic supplements could enhance recovery and protect piglets against enteric pathogens. Seven local lactic acid bacteria (LAB), (Ent. faecium 79N and 40N, Lact. plantarum 22F, 25F and 31F, Ped. acidilactici 72N and Ped. pentosaceus 77F) from pig feces were well-characterized as high potential probiotics. Cell-free supernatants (CFS) and live LAB were evaluated for antiviral activities by co-incubation on Vero cells and challenged with a pandemic strain of PEDV isolated from pigs in Thailand. Cell survival and viral inhibition were determined by cytopathic effect (CPE) reduction assay and confirmed by immunofluorescence. At 1:16, CFS dilution (pH 6.3–6.8) showed no cytotoxicity in Vero cells and was therefore used as the dilution for antiviral assays. The diluted CFS of all Lact. plantarum showed the antiviral effect against PEDV; however, the same antiviral effect could not be observed in Ent. faecium and Pediococcus strains. In competitive experiment, only live Lact. plantarum 25F and Ped. pentosaceus 77F showed CPE reduction in the viral infected cells to <50% observed field area. This study concluded that the CFS of all tested lactobacilli, and live Lact. plantarum (22F and 25F) and Pediococcus strains 72N and 77F could reduce infectivity of the pandemic strain of PEDV from pigs in Thailand on the target Vero cells.

Keywords

Antiviral activity Cell-free supernatants Lactic acid bacteria Porcine epidemic diarrhea virus Probiotics 

Notes

Acknowledgements

This study was financially supported by The 90th Anniversary of Chulalongkorn University Fund (Ratchadaphiseksomphot Endowment Fund), STAR; Diagnosis and Monitoring of Animal Pathogen, Chulalongkorn University and the Agricultural Research Development Agency (ARDA) (No. 5803090002) (Public Organization), Thailand. We thank Professor Somboon Tanasupawat for assistance and comments to improve the manuscript.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12602_2017_9281_MOESM1_ESM.docx (18 kb)
ESM 1 (DOCX 17 kb)

References

  1. 1.
    Shen H, Zhang C, Guo P, Liu Z, Zhang J (2015) Effective inhibition of porcine epidemic diarrhea virus by RNA interference in vitro. Virus Genes 51(2):252–259CrossRefGoogle Scholar
  2. 2.
    Lee C (2015) Porcine epidemic diarrhea virus: an emerging and re-emerging epizootic swine virus. Virol J 12(1):193CrossRefGoogle Scholar
  3. 3.
    Di-qiu L, Jun-wei G, Xin-yuan Q, Yan-ping J, Song-mei L, Yi-jing L (2012) High-level mucosal and systemic immune responses induced by oral administration with lactobacillus-expressed porcine epidemic diarrhea virus (PEDV) S1 region combined with lactobacillus-expressed N protein. Appl Microbiol Biot 93(6):2437–2446CrossRefGoogle Scholar
  4. 4.
    Acheson DW, Luccioli S (2004) Microbial-gut interactions in health and disease. Mucosal immune responses. Best Pract Res Cl Ga 18(2):387–404. doi: 10.1016/j.bpg.2003.11.002 CrossRefGoogle Scholar
  5. 5.
    Kaila M, Isolauri E, Saxelin M, Arvilommi H, Vesikari T (1995) Viable versus inactivated lactobacillus strain GG in acute rotavirus diarrhoea. Arch Dis Child 72(1):51–53CrossRefGoogle Scholar
  6. 6.
    Cross ML (2002) Immunoregulation by probiotic lactobacilli: pro-Th1 signals and their relevance to human health. Clin Appl Immunol Rev 3(3):115–125CrossRefGoogle Scholar
  7. 7.
    Tannock GW (1997) Probiotic properties of lactic-acid bacteria: plenty of scope for fundamental R & D. Trends Biotechnol 15(7):270–274CrossRefGoogle Scholar
  8. 8.
    Choi H-J, Song J-H, Ahn Y-J, Baek S-H, Kwon D-H (2009) Antiviral activities of cell-free supernatants of yogurts metabolites against some RNA viruses. Eur Food Res Technol 228(6):945–950CrossRefGoogle Scholar
  9. 9.
    Chang TL-Y, Chang C-H, Simpson DA, Xu Q, Martin PK, Lagenaur LA, Schoolnik GK, Ho DD, Hillier SL, Holodniy M (2003) Inhibition of HIV infectivity by a natural human isolate of lactobacillus jensenii engineered to express functional two-domain CD4. P Natl A Sci 100(20):11672–11677CrossRefGoogle Scholar
  10. 10.
    Isolauri E (2003) Probiotics for infectious diarrhoea. Gut 52(3):436–437CrossRefGoogle Scholar
  11. 11.
    Chai W, Burwinkel M, Wang Z, Palissa C, Esch B, Twardziok S, Rieger J, Wrede P, Schmidt MFG (2012) Antiviral effects of a probiotic Enterococcus faecium strain against transmissible gastroenteritis coronavirus. Arch Virol 158(4):799–807. doi: 10.1007/s00705-012-1543-0 CrossRefGoogle Scholar
  12. 12.
    Botić T, Danø T, Weingartl H, Cencič A (2007) A novel eukaryotic cell culture model to study antiviral activity of potential probiotic bacteria. Int J Food Microbiol 115(2):227–234CrossRefGoogle Scholar
  13. 13.
    Maragkoudakis PA, Chingwaru W, Gradisnik L, Tsakalidou E, Cencic A (2010) Lactic acid bacteria efficiently protect human and animal intestinal epithelial and immune cells from enteric virus infection. Int J Food Microbiol 141:S91–S97CrossRefGoogle Scholar
  14. 14.
    Hofmann M, Wyler R (1988) Propagation of the virus of porcine epidemic diarrhea in cell culture. J Clin Microbiol 26(11):2235–2239Google Scholar
  15. 15.
    Reed L, Muench H (1938) A simple method of estimating fifty percent endpoints. Am J Hyg 27:493–497Google Scholar
  16. 16.
    Wang C-Y, Lin P-R, Ng C-C, Shyu Y-T (2010) Probiotic properties of Lactobacillus strains isolated from the feces of breast-fed infants and Taiwanese pickled cabbage. Anaerobe 16(6):578–585. doi: 10.1016/j.anaerobe.2010.10.003 CrossRefGoogle Scholar
  17. 17.
    Lin W-H, Hwang C-F, Chen L-W, Tsen H-Y (2006) Viable counts, characteristic evaluation for commercial lactic acid bacteria products. Food Microbiol 23(1):74–81CrossRefGoogle Scholar
  18. 18.
    Borenfreund E, Puerner JA (1985) Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicol Lett 24(2):119–124CrossRefGoogle Scholar
  19. 19.
    Seo BJ, Mun MR, Kim C-J, Lee I, Kim H, Park Y-H (2010) Putative probiotic Lactobacillus spp. from porcine gastrointestinal tract inhibit transmissible gastroenteritis coronavirus and enteric bacterial pathogens. Trop Anim Health Pro 42(8):1855–1860CrossRefGoogle Scholar
  20. 20.
    Fayol-Messaoudi D, Berger CN, Coconnier-Polter M-H, Lievin-Le Moal V, Servin AL (2005) pH-, lactic acid-, and non-lactic acid-dependent activities of probiotic Lactobacilli against Salmonella enterica serovar Typhimurium. Appl Environ Microbiol 71(10):6008–6013CrossRefGoogle Scholar
  21. 21.
    Ermolenko E, Furaeva V, Isakov V, Ermolenko D, Suvorov A (2009) Inhibition of herpes simplex virus type 1 reproduction by probiotic bacteria in vitro. Vopr Virusol 55(4):25–28Google Scholar
  22. 22.
    Saarela M, Mogensen G, Fonden R, Mättö J, Mattila-Sandholm T (2000) Probiotic bacteria: safety, functional and technological properties. J Biotech 84(3):197–215CrossRefGoogle Scholar
  23. 23.
    Dembinski JL, Hungnes O, Hauge AG, Kristoffersen A-C, Haneberg B, Mjaaland S (2014) Hydrogen peroxide inactivation of influenza virus preserves antigenic structure and immunogenicity. J Virol Methods 207:232–237CrossRefGoogle Scholar
  24. 24.
    Yang S-C, Lin C-H, Sung CT, Fang J-Y (2014) Antibacterial activities of bacteriocins: application in foods and pharmaceuticals. Front Microbiol 5:241Google Scholar
  25. 25.
    Aboubakr HA, El-Banna AA, Youssef MM, Al-Sohaimy SA, Goyal SM (2014) Antiviral effects of Lactococcus lactis on feline calicivirus, a human norovirus surrogate. Food Environ Virol 6(4):282–289CrossRefGoogle Scholar
  26. 26.
    Allayeh AK, Dardeer EE, Kotb NS (2015) Effects of cell-free supernatants of yogurts metabolites on Coxsackie B3 virus in vitro and in vivo. Middle East J Appl Sci 5:353–358Google Scholar
  27. 27.
    Charteris W, Kelly P, Morelli L, Collins J (1998) Development and application of an in vitro methodology to determine the transit tolerance of potentially probiotic Lactobacillus and Bifidobacterium species in the upper human gastrointestinal tract. J Appl Microbiol 84(5):759–768CrossRefGoogle Scholar
  28. 28.
    Lee Y, Lim C, Teng W, Ouwehand A, Tuomola E, Salminen S (2000) Quantitative approach in the study of adhesion of lactic acid bacteria to intestinal cells and their competition with enterobacteria. Appl Environ Microbiol 66(9):3692–3369CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Wandee Sirichokchatchawan
    • 1
  • Gun Temeeyasen
    • 1
  • Dachrit Nilubol
    • 1
  • Nuvee Prapasarakul
    • 1
  1. 1.Faculty of Veterinary Science, Department of Veterinary MicrobiologyChulalongkorn UniversityBangkokThailand

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