Isolation and Identification of Lactic Acid Bacteria Inhibiting Gastro-intestinal Pathogenic Bacteria of Domestic Animal.

가축 소화기 병원성 세균을 저해하는 유산균의 분리 및 동정

  • Published : 2002.06.01

Abstract

To isolate probiotic lactic acid bacteria having superior inhibitory activities against animal gastro-intestinal pathogenic bacteria such as Salmonella gallinarum, Staphylococcus aureus and Escherichia coli, 130 strains were initially isolated from the small intestines of Korean native chickens and 7 lactic acid bacteria were finally selected. By using API CHL kit and 16S rRNA sequencing method, the selected lactic acid bacteria were found to be belonged to genus Lactobacillus except BD14 identified as Pediococcus pentosaceus. Especially, Lactobacillus pentosus K34 showed the highest resistancy to both of HCl and bile salt, as well as the highest inhibitory activities against S. gallinarum, S. aureus and E. coli. All the selected strains were sensitive to various antibiotics such as neomycin, erythromycin, cephalosporin, amoxicillin/clavulanic acid, ampicillin, oxytetracycline, but resistant to ciprofloxacin. All the selected strains except BL strain were resistant to colistin and streptomycin, and BD14, BD16, K34 strains were resistant to gentamicin.

토종닭 소장으로부터 가축 소화기 병원성 유해세균의 분리를 위하여 무작위 선별법과 중층-도말법을 사용하여 총130주의 유산균을 1차 분리하였다. 1차 분리한 130주를 paper disc법을 사용하여 최종적으로 살모넬라 균, 포도상구균, 대장균에 대하여 저해능이 우수한 7주를 분리하였다. 분리 균주의 동정을 위하여 1차로 API CHL kit를 사용하였고, 동정의 정확성을 높이기 위하여 2차로 16S rRNA sequencing 방법을 사용하였다. 그 결과 BD14 균주는 Pediococcus pentosaceus로 동정되었으며 나머지 6주는 Lactobacillus sp.로 동정되어 모두 안전성 있는 유산균임을 확인하였다. 여러 병원균에 가장 항균력이 큰 유산균은 L. pentosus K34이었으며, 이 유산균은 염산과 담즙산에 대한 내성에서도 가장 우수한 균주로 나타났다. 10종의 항생제에 대한 감수성을 조사한 결과 ciprofloxacin에 대하여 모든 균주가 내성을 보였고, BL 균주를 제외하고 모든 균주가 colistin, streptomycin에 대하여 내성을 보였다. 또한 BD14, BD16, K34 균주가 gentamicin에 대한 내성을 보였다.

Keywords

References

  1. Kor. J. Appl. Microbiol. Biotechnol v.28 Isolation and characterization of Lactobacillus fermentum YL-3 as poultry probiotics Cho, M. K.;K. Kim;C. H. Kim;T. K. Lee;K. Y. Kim
  2. Br. Vet. J. v.143 Efficacy and financial value of antibiotictreatment of bovine clinical mastitis during lactation-a Review Craven, N. https://doi.org/10.1016/0007-1935(87)90018-2
  3. ASM News v.66 Antibiotics as growth promoters : Risks and alternatives Dixon, B.
  4. Preweaning colibacillosis In APSA Comittee(ed.), Manipulating pig production Fahy, V. A.;D. Connaughton;S. J. Dresen;E. M. Spicer
  5. Selection of strains for probiotic use The scientific basis Havenaar, R.;B. T. Brink;J. H. J. I. Veid;R. Fuller(ed.)
  6. Can. J. Comp. Med. v.47 Neonatal diarrhea of pig in Quebec : infectious causes of significant outbreaks Morin, M.;D. Turgeon;J. Jolette;Y. Robinson;J. B. Phaneuf;M. Sauvageau;E. Teuscher;R. Higgins;S. Larivi'ere
  7. Kor. J. Appl. Microbiol. Biotechnol. v.28 Prevalence of anti-biotic-resistant strains among bacteria isolated from bovine mastitis, swine diarrhea, and swine pneumonia Park, J. C.;I. S. Kim;S. K. Kwon;J. M. Noh;S. M. Lee;J. P. Park;W. K. Lee;S. R. Ryu
  8. Diseases of poultry Fowl typhoid Pomeroy, B. S.;K. V. Nagaraja;B. W. Calnek(eds.);H. J. Barnes(eds.);C. W. Beard(eds.);W. M. Reid(eds.);H. W. Yoder(eds.)
  9. J. Nutr. v.130 The role of probiotic cultures in the control of gastrointestinal health Rolfe, R. D.
  10. J. Nutr. v.130 Considerations for use of probiotic bacteria to modulate human health Sanders, M. E.
  11. Microbiology v.147 Characterization of lactobacilli from Scotch malt whisky distilleries and description of Lactobacillus ferintoshensis sp. nov., a new species isolated from malt whisky fermentations Simpson K. L.;B. Pettersson;F. G. Priest https://doi.org/10.1099/00221287-147-4-1007
  12. Human Intestinal Microflora in Health and Disease Effect of dietary and environmental stress on the gastrointestinal microbiota Tannock, G. W.;D. J. Hentges(ed.)
  13. Compendium of methods for the microbiological examination of foods, 3th ed. Vanderzant, C.;D. F. Splittstoesser
  14. Probiotics : The scientific basis Probiotics for ruminants Wallace, R. J.;C. J. Newbold;R. Fuller(ed.)