Czech J. Food Sci., 2021, 39(4):305-311 | DOI: 10.17221/296/2020-CJFS

Isolation of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus from nature: Technological characterisation and antibiotic resistanceOriginal Paper

Hakan Tavsanli ORCID...*,1, Tülay Elal Mus2, Figen Cetinkaya3, Ergün Ayanoglu3, Recep Cibik3
1 Department of Veterinary Public Health, Faculty of Veterinary Medicine, Balikesir University, Balikesir, Turkey
2 Department of Food Processing, Vocational School of Keles, Bursa Uludag University, Keles, Turkey
3 Department of Food Hygiene and Technology, Faculty of Veterinary Medicine, Bursa Uludag University, Bursa, Turkey

Yoghurt fermenting bacteria were isolated from natural sources including plants, dew, and rain samples (total of 300 samples) by the same methods nomadic peoples used for several centuries in Turkey. Inoculation into the reconstituted skim milk followed by planting on specific media and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) analysis allowed for the identification of 18 Lactobacillus delbrueckii subsp. and 26 Streptococcus thermophilus. A multiplex polymerase chain reaction (PCR) assay applied to lactobacilli enabled the identification of 5 isolates as L. delbrueckii subsp. bulgaricus. The isolates showed a varying range of acidification rates and proteolytic activity in reconstituted skimmed milk (RSM). S. thermophilus isolates showed a broader range of resistance and the most frequent resistance was observed for streptomycin (69.2%), gentamycin (65.3%), clindamycin (61.5%), ampicillin (61.5%), kanamycin (53.8%), and erythromycin (50%). For L. delbrueckii subsp. the highest resistance was determined for vancomycin (38.8%), ciprofloxacin (33.3%), and penicillin (27.8%). The frequency of multiple resistance was tested on 14 different antimicrobials determining that 19 S. thermophilus (73%) and 3 L. delbrueckii subsp. (16.7%) demonstrated resistance to more than three different antibiotics. In contrast to this wide-ranging resistance, five isolates from each genus were found to be susceptible to all tested antibiotics. The present study indicates that lactic acid bacteria (LAB) isolated from nature may have broad-range of resistance to antibiotics and could be a source for the transfer of resistance.

Keywords: yoghurt bacteria; natural isolates; characterisation; antibiotic resistance

Published: August 29, 2021  Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Tavsanli H, Elal Mus T, Cetinkaya F, Ayanoglu E, Cibik R. Isolation of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus from nature: Technological characterisation and antibiotic resistance. Czech J. Food Sci.. 2021;39(4):305-311. doi: 10.17221/296/2020-CJFS.
Download citation

References

  1. Akpinar A., Yerlikaya O., Kiliç S. (2011): Antimicrobial activity and antibiotic resistance of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus strains isolated from Turkish homemade yoghurts. African Journal of Microbiology Research, 5: 675-682.
  2. Bostan K., Unver Alcay A., Yalçin S., Vapur U.E., Nizamoglu M. (2017): Identification and characterisation of lactic acid bacteria isolated from traditional cone yoghurt. Food Science and Biotechnology, 26: 1625-1632. Go to original source... Go to PubMed...
  3. Cibik R., Cetinkaya F., Ersoy M., Yibar A. (2010): Identification and technological characterisation of Lactococcus isolated from traditional Turkish cheeses. Revue de Medecine Veterinaire, 161: 509-514.
  4. Cogan T.M., Barbosa M., Beuvier E., Bianchi-Salvadori B.R.U.N.A., Cocconcelli P.S., Fernandes I., Medina M. (1997): Characterisation of the lactic acid bacteria in artisanal dairy products. Journal of Dairy Research, 64: 409-421. Go to original source...
  5. Cremonesi P., Vanoni L., Morandi S., Silvetti T., Castiglioni B., Brasca M. (2011): Development of a pentaplex PCR assay for the simultaneous detection of Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. helveticus, L. fermentum in whey starter for Grana Padano cheese. International Journal of Food Microbiology, 146: 207-221. Go to original source... Go to PubMed...
  6. Crowley S., Mahony J., Van Sinderen D. (2013): Current perspectives on antifungal lactic acid bacteria as natural bio-preservatives. Trends in Food Science and Technology, 33: 93-109. Go to original source...
  7. D'Aimmo M.R., Modesto M., Biavati B. (2007): Antibiotic resistance of lactic acid bacteria and Bifidobacterium spp. isolated from dairy and pharmaceutical products. International Journal of Food Microbiology, 115: 35-42. Go to original source... Go to PubMed...
  8. Dimitrov Z., Michaylova M., Mincova S. (2005): Characterisation of Lactobacillus helveticus strains isolated from Bulgarian yoghurt, cheese, plants and human faecal samples by sodium dodecilsulfate polyacrylamide gel electrophoresis of cell-wall proteins, ribotyping and pulsed field gel fingerprinting. International Dairy Journal, 15: 998-1005. Go to original source...
  9. Driessen F.M., Kingma F., Stadhouders J. (1982): Evidence that Lactobacillus bulgaricus in yogurt is stimulated by carbon dioxide produced by Streptococcus thermophilus. Netherlands Milk and Dairy Journal, 36: 135-144.
  10. EFSA (2012): European Food Safety Authority (EFSA) Panel on Additives and Products or Substances used in Animal Feed (FEEDAP): Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA Journal, 10: 2740-2750. Go to original source...
  11. Erkus O., Okuklu B., Yenidunya A.F., Harsa S. (2014): High genetic and phenotypic variability of Streptococcus thermophilus strains isolated from artisanal Yuruk yoghurts. LWT - Food Science and Technology, 58: 348-354. Go to original source...
  12. Georgieva R., Yocheva L., Tserovska L., Zhelezova G., Stefanova N., Atanasova A., Karaivanova E. (2015): Antimicrobial activity and antibiotic susceptibility of Lactobacillus and Bifidobacterium spp. intended for use as starter and probiotic cultures. Biotechnology and Biotechnological Equipment, 29: 84-91. Go to original source... Go to PubMed...
  13. Karapetkov N., Georgieva R., Rumyan N., Karaivanova E. (2011): Antibiotic susceptibility of different lactic acid bacteria strains. Beneficial Microbes, 2: 335-339. Go to original source... Go to PubMed...
  14. Klein G., Hallmann C., Casas I.A., Abad J., Louwers J., Reuter G. (2000): Exclusion of van A, van B and van C type glycopeptide resistance in strains of Lactobacillus reuteri and Lactobacillus rhamnosus used as probiotics by polymerase chain reaction and hybridisation methods. Journal of Applied Microbiology, 89: 815-824. Go to original source... Go to PubMed...
  15. Mathur S., Singh R. (2005): Antibiotic resistance in food lactic acid bacteria - A review. International Journal of Food Microbiology, 105: 281-295. Go to original source... Go to PubMed...
  16. Michaylova M., Minkova S., Kimura K., Sasaki T., Isawa K. (2007): Isolation and characterisation of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus from plants in Bulgaria. FEMS Microbiology Letters, 269: 160-169. Go to original source... Go to PubMed...
  17. Morandi S., Silvetti T., Miranda Lopez J.M., Brasca M. (2015): Antimicrobial activity, antibiotic resistance and the safety of lactic acid bacteria in raw milk Valtellina Casera cheese. Journal of Food Safety, 35: 193-205. Go to original source...
  18. Nawaz M., Wang J., Zhou A., Ma C., Wu X., Moore J.E., Xu J. (2011): Characterisation and transfer of antibiotic resistance in lactic acid bacteria from fermented food products. Current Microbiology, 62: 1081-1089. Go to original source... Go to PubMed...
  19. Salvucci E., LeBlanc J.G., Pérez G. (2016): Technological properties of lactic acid bacteria isolated from raw cereal material. LWT - Food Science and Technology, 70: 185-191. Go to original source...
  20. Sherman J. M., (1937): The streptococci. Bacteriology Reviews, 1: 3-97. Go to original source... Go to PubMed...
  21. Tang H., Yuan J., Xie C., Wei H. (2007): Antibiotic susceptibility of strains in Chinese medical probiotic products. Journal of Medical Colleges of PLA, 22: 149-152. Go to original source...
  22. Urshev Z., Ninova-Nikolova N., Ishlimova D., PashovaBaltova K., Michaylova M., Savova T. (2014): Selection and characterisation of naturally occurring high acidification rate Streptococcus thermophilus strains. Biotechnology and Biotechnological Equipment, 25: 899-903. Go to original source... Go to PubMed...
  23. Wang K., Zhang H., Feng J., Ma L., de la Fuente-Núñez C., Wang S., Lu X. (2019): Antibiotic resistance of lactic acid bacteria isolated from dairy products in Tianjin, China. Journal of Agriculture and Food Research, 1: 1-5. Go to original source...
  24. Weinrichter B., Luginbühl W., Rohm H., Jimeno J. (2001): Differentiation of facultatively heterofermentative lactobacilli from plants, milk, and hard type cheeses by SDS-PAGE, RAPD, FTIR, energy source utilization and autolysis type. Lebensmittel-Wissenschaft and Technologie, 34: 556-566. Go to original source...
  25. Xu Z., Li S., Gong G., Liu Z., Wu Z., Ma C. (2015): Influence of different acidifying strains of Lactobacillus delbrueckii subsp. bulgaricus on the quality of yoghurt. Food Science and Technology Research, 21: 263-269. Go to original source...
  26. Zhang H., Xie L., Zhang W., Zhou W., Su J., Liu J. (2013): The association of biofilm formation with antibiotic resistance in lactic acid bacteria from fermented foods. Journal of Food Safety, 33: 114-120. Go to original source...
  27. Zhou N., Zhang J.X., Fan M.T., Wang J., Guo G., Guo G., Wei X.Y. (2012): Antibiotic resistance of lactic acid bacteria isolated from Chinese yogurts. Journal of Dairy Science, 95: 4775-4783. Go to original source... Go to PubMed...

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.