The phenotypic and genotypic antibiotic susceptibility of vaginal Lactobacillus with potential probiotic properties isolated from healthy women in northern Iran
,
Abstract
Background and Objectives: This study assesses the antibiotic susceptibility of vaginal Lactobacillus strains and provides data for determining the prevalence of certain antibiotic resistance genes in the new strains of lactobacilli serving as probiotics and selected from healthy women in northern Iran.
Materials and Methods: One hundred premenopausal non-pregnant women in the reproductive age range of 22-50 years participated in this study. The potential probiotic vaginal lactobacilli used in the study included Lactobacillus crispatus (34.2%), Lactobacillus gasseri (26.3%), Lactobacillus johnsonii (10.5%), Lactobacillus acidophilus (15.7%) and Lactobacillus jensenii (13.1%). The phenotypic antibiotic susceptibility of the strains was determined by E test and DNA extraction and PCR were performed to examine the antibiotic resistance genes.
Results: 38 potential probiotic vaginal lactobacilli were isolated. All the strains of lactobacilli were resistant to metronidazole and trimethoprim/sulfamethoxazole and all of the strains were susceptible to ampicillin and chloramphenicol antibiotics. The results showed that ermB, ermC, and ermA genes were observed in the strains of Lactobacillus acidophilus. Metronidazole resistance (nim) gene was also found in one strain of Lactobacillus crispatus and Lactobacillus johnsonii. The aminoglycoside resistance (aac6'-aph2") gene was observed in 8% of the strains. Also, tetM, tetK and tetW genes were found in more than 80% of the Lactobacillus strains.
Conclusion: The antimicrobial susceptibility of vaginal lactobacilli is an important criterion for establishing whether or not the organism is a probiotic. A high level of resistance to clinical antibiotics, such as metronidazole and aminoglycosides, was demonstrated. Antibiotic resistant genes also appeared widely in vaginal lactobacilli.
1. Choudhury R, Panda S, Singh DV. Emergence and dissemination of antibiotic resistance: a global problem. Indian J Med Microbiol 2012; 30: 384-390.
2. Machowska A, Stålsby Lundborg C. Drivers of irrational use of antibiotics in Europe. Int J Environ Res Public Health 2018; 16: 27.
3. Singer RS, Finch R, Wegener HC, Bywater R, Walters J, Lipsitch M. Antibiotic resistance--the interplay between antibiotic use in animals and human beings. Lancet Infect Dis 2003; 3: 47-51.
4. Mathur S, Singh R. Antibiotic resistance in food lactic acid bacteria--a review. Int J Food Microbiol 2005; 105: 281-295.
5. Fraqueza MJ. Antibiotic resistance of lactic acid bacteria isolated from dry-fermented sausages. Int J Food Microbiol 2015; 212: 76-88.
6. Sharma P, Tomar SK, Goswami P, Sangwan V, Singh R. Antibiotic resistance among commercially available probiotics. Food Res Int 2014; 57: 176-195.
7. Georgieva R, Yocheva L, Tserovska L, Zhelezova G, Stefanova N, Atanasova A, et al. Antimicrobial activity and antibiotic susceptibility of Lactobacillus and Bifidobacterium spp. intended for use as starter and probiotic cultures. Biotechnol Biotechnol Equip 2015; 29: 84-91.
8. Allahverdiyev AM, Kon KV, Abamor ES, Bagirova M, Rafailovich M. Coping with antibiotic resistance: combining nanoparticles with antibiotics and other antimicrobial agents. Expert Rev Anti Infect Ther 2011; 9: 1035-1052.
9. Anisimova EA, Yarullina DR. Antibiotic resistance of Lactobacillus strains. Curr Microbiol 2019; 76: 1407-1416.
10. Gueimonde M, Sánchez B, G de Los Reyes-Gavilán C, Margolles A. Antibiotic resistance in probiotic bacteria. Front Microbiol 2013; 4: 202.
11. Ammor MS, Flórez AB, van Hoek AH, de Los Reyes-Gavilán CG, Aarts HJ, Margolles A, et al. Molecular characterization of intrinsic and acquired antibiotic resistance in lactic acid bacteria and bifidobacteria. J Mol Microbiol Biotechnol 2008; 14: 6-15.
12. Dimitonova SP, Danova ST, Serkedjieva JP, Bakalov BV. Antimicrobial activity and protective properties of vaginal lactobacilli from healthy Bulgarian women. Anaerobe 2007; 13: 178-184.
13. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP); Rychen G, Aquilina G, Azimonti G, Bampidis V, Bastos ML, Bories G, et al. Guidance on the characterisation of microorganisms used as feed additives or as production organisms. EFSA J 2018; 16(3): e05206.
14. Frieri M, Kumar K, Boutin A. Antibiotic resistance. J Infect Public Health 2017; 10: 369-378.
15. Argyri AA, Zoumpopoulou G, Karatzas KA, Tsakalidou E, Nychas GJ, Panagou EZ, et al. Selection of potential probiotic lactic acid bacteria from fermented olives by in vitro tests. Food Microbiol 2013; 33: 282-291.
16. Zhou JS, Pillidge CJ, Gopal PK, Gill HS. Antibiotic susceptibility profiles of new probiotic Lactobacillus and Bifidobacterium strains. Int J Food Microbiol 2005; 98: 211-217.
17. Kaewnopparat S, Dangmanee N, Kaewnopparat N, Srichana T, Chulasiri M, Settharaksa S. Inávitro probiotic properties of Lactobacillus fermentum SK5 isolated from vagina ofáa healthy woman. Anaerobe 2013; 22: 6-13.
18. Nami Y, Haghshenas B, Yari Khosroushahi A. Molecular identification and probiotic potential characterization of lactic acid bacteria isolated from human vaginal microbiota. Adv Pharm Bull 2018; 8: 683-695.
19. Bernardeau M, Vernoux JP, Henri-Dubernet S, Guéguen M. Safety assessment of dairy microorganisms: the Lactobacillus genus. Int J Food Microbiol 2008; 126: 278-285.
20. Simoes JA, Aroutcheva AA, Shott S, Faro S. Effect of metronidazole on the growth of vaginal lactobacilli in vitro. Infect Dis Obstet Gynecol 2001; 9: 41-45.
21. Štšepetova J, Taelma H, Smidt I, Hütt P, Lapp E, Aotäht E, et al. Assessment of phenotypic and genotypic antibiotic susceptibility of vaginal Lactobacillus sp. J Appl Microbiol 2017; 123: 524-534.
22. Pino A, Bartolo E, Caggia C, Cianci A, Randazzo CL. Detection of vaginal lactobacilli as probiotic candidates. Sci Rep 2019; 9: 3355.
23. Morroni G, Brenciani A, Litta-Mulondo A, Vignaroli C, Mangiaterra G, Fioriti S, et al. Characterization of a new transferable MDR plasmid carrying the pbp5 gene from a clade B commensal Enterococcus faecium. J Antimicrob Chemother 2019; 74: 843-850.
24. Hütt P, Lapp E, Štšepetova J, Smidt I, Taelma H, Borovkova N, et al. Characterisation of probiotic properties in human vaginal lactobacilli strains. Microb Ecol Health Dis 2016; 27: 30484.
25. Danielsen M, Wind A. Susceptibility of Lactobacillus spp. to antimicrobial agents. Int J Food Microbiol 2003; 82: 1-11.
26. Hoque MZ, Akter F, Hossain KM, Rahman MSM, Billah MM, Islam KMD. Isolation, identification and analysis of probiotic properties of Lactobacillus spp. from selective regional yoghurts. World J Dairy Food Sci 2010; 5: 39-46.
27. Devirgiliis C, Zinno P, Perozzi G. Update on antibiotic resistance in foodborne Lactobacillus and Lactococcus species. Front Microbiol 2013; 4: 301.
28. Rigo GV, Tasca T. Vaginitis: Review on drug resistance. Curr Drug Targets 2020; 21: 1672-1686.
29. Kiray E, Kariptas E, Azarkan SY. Evaluation of vaginal lactobacilli with potential probiotic properties and biotherapeutic effects isolated from healthy Turkish women. PSM Microbiol 2019; 4: 56-70.
30. Abriouel H, Casado Muñoz MDC, Lavilla Lerma L, Pérez Montoro B, Bockelmann W, Pichner R, et al. New insights in antibiotic resistance of Lactobacillus species from fermented foods. Food Res Int 2015; 78: 465-481.
31. Sirichoat A, Flórez AB, Vázquez L, Buppasiri P, Panya M, Lulitanond V, et al. Antibiotic resistance-susceptibility profiles of Enterococcus faecalis and Streptococcus spp. from the human vagina, and genome analysis of the genetic basis of intrinsic and acquired resistances. Front Microbiol 2020; 11: 1438.
32. Kassaa AI, Hamze M, Hober D, Chihib NE, Drider D. Identification of vaginal lactobacilli with potential probiotic properties isolated from women in North Lebonan. Microb Ecol 2014;67:722-734.
33. Hummel AS, Hertel C, Holzapfel WH, Franz CM. Antibiotic resistances of starter and probiotic strains of lactic acid bacteria. Appl Environ Microbiol 2007; 73: 730-739.
34. Sirichoat A, Flórez AB, Vázquez L, Buppasiri P, Panya M, Lulitanond V, et al. Antibiotic susceptibility profiles of lactic acid bacteria from the human vagina and genetic basis of acquired resistances. Int J Mol Sci 2020; 21: 2594.
35. Lim KT, Hanifah YA, Yusof M, Thong KL. ermA, ermC, tetM and tetK are essential for erythromycin and tetracycline resistance among methicillin-resistant Staphylococcus aureus strains isolated from a tertiary hospital in Malaysia. Indian J Med Microbiol 2012; 30: 203-207.
36. Begovic J, Huys G, Mayo B, D'Haene K, Florez AB, Lozo J, et al. Human vaginal Lactobacillus rhamnosus harbor mutation in 23S rRNA associated with erythromycin resistance. Res Microbiol 2009; 160: 421-426.
37. Goossens H. Assessing the safety of probiotics with regard to antibiotic resistance, in Functional dairy products. 2007, Elsevier. p. 427-442.
38. Delgado S, Flórez AB, Mayo B. Antibiotic susceptibility of Lactobacillus and Bifidobacterium species from the human gastrointestinal tract. Curr Microbiol 2005; 50: 202-207.
2. Machowska A, Stålsby Lundborg C. Drivers of irrational use of antibiotics in Europe. Int J Environ Res Public Health 2018; 16: 27.
3. Singer RS, Finch R, Wegener HC, Bywater R, Walters J, Lipsitch M. Antibiotic resistance--the interplay between antibiotic use in animals and human beings. Lancet Infect Dis 2003; 3: 47-51.
4. Mathur S, Singh R. Antibiotic resistance in food lactic acid bacteria--a review. Int J Food Microbiol 2005; 105: 281-295.
5. Fraqueza MJ. Antibiotic resistance of lactic acid bacteria isolated from dry-fermented sausages. Int J Food Microbiol 2015; 212: 76-88.
6. Sharma P, Tomar SK, Goswami P, Sangwan V, Singh R. Antibiotic resistance among commercially available probiotics. Food Res Int 2014; 57: 176-195.
7. Georgieva R, Yocheva L, Tserovska L, Zhelezova G, Stefanova N, Atanasova A, et al. Antimicrobial activity and antibiotic susceptibility of Lactobacillus and Bifidobacterium spp. intended for use as starter and probiotic cultures. Biotechnol Biotechnol Equip 2015; 29: 84-91.
8. Allahverdiyev AM, Kon KV, Abamor ES, Bagirova M, Rafailovich M. Coping with antibiotic resistance: combining nanoparticles with antibiotics and other antimicrobial agents. Expert Rev Anti Infect Ther 2011; 9: 1035-1052.
9. Anisimova EA, Yarullina DR. Antibiotic resistance of Lactobacillus strains. Curr Microbiol 2019; 76: 1407-1416.
10. Gueimonde M, Sánchez B, G de Los Reyes-Gavilán C, Margolles A. Antibiotic resistance in probiotic bacteria. Front Microbiol 2013; 4: 202.
11. Ammor MS, Flórez AB, van Hoek AH, de Los Reyes-Gavilán CG, Aarts HJ, Margolles A, et al. Molecular characterization of intrinsic and acquired antibiotic resistance in lactic acid bacteria and bifidobacteria. J Mol Microbiol Biotechnol 2008; 14: 6-15.
12. Dimitonova SP, Danova ST, Serkedjieva JP, Bakalov BV. Antimicrobial activity and protective properties of vaginal lactobacilli from healthy Bulgarian women. Anaerobe 2007; 13: 178-184.
13. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP); Rychen G, Aquilina G, Azimonti G, Bampidis V, Bastos ML, Bories G, et al. Guidance on the characterisation of microorganisms used as feed additives or as production organisms. EFSA J 2018; 16(3): e05206.
14. Frieri M, Kumar K, Boutin A. Antibiotic resistance. J Infect Public Health 2017; 10: 369-378.
15. Argyri AA, Zoumpopoulou G, Karatzas KA, Tsakalidou E, Nychas GJ, Panagou EZ, et al. Selection of potential probiotic lactic acid bacteria from fermented olives by in vitro tests. Food Microbiol 2013; 33: 282-291.
16. Zhou JS, Pillidge CJ, Gopal PK, Gill HS. Antibiotic susceptibility profiles of new probiotic Lactobacillus and Bifidobacterium strains. Int J Food Microbiol 2005; 98: 211-217.
17. Kaewnopparat S, Dangmanee N, Kaewnopparat N, Srichana T, Chulasiri M, Settharaksa S. Inávitro probiotic properties of Lactobacillus fermentum SK5 isolated from vagina ofáa healthy woman. Anaerobe 2013; 22: 6-13.
18. Nami Y, Haghshenas B, Yari Khosroushahi A. Molecular identification and probiotic potential characterization of lactic acid bacteria isolated from human vaginal microbiota. Adv Pharm Bull 2018; 8: 683-695.
19. Bernardeau M, Vernoux JP, Henri-Dubernet S, Guéguen M. Safety assessment of dairy microorganisms: the Lactobacillus genus. Int J Food Microbiol 2008; 126: 278-285.
20. Simoes JA, Aroutcheva AA, Shott S, Faro S. Effect of metronidazole on the growth of vaginal lactobacilli in vitro. Infect Dis Obstet Gynecol 2001; 9: 41-45.
21. Štšepetova J, Taelma H, Smidt I, Hütt P, Lapp E, Aotäht E, et al. Assessment of phenotypic and genotypic antibiotic susceptibility of vaginal Lactobacillus sp. J Appl Microbiol 2017; 123: 524-534.
22. Pino A, Bartolo E, Caggia C, Cianci A, Randazzo CL. Detection of vaginal lactobacilli as probiotic candidates. Sci Rep 2019; 9: 3355.
23. Morroni G, Brenciani A, Litta-Mulondo A, Vignaroli C, Mangiaterra G, Fioriti S, et al. Characterization of a new transferable MDR plasmid carrying the pbp5 gene from a clade B commensal Enterococcus faecium. J Antimicrob Chemother 2019; 74: 843-850.
24. Hütt P, Lapp E, Štšepetova J, Smidt I, Taelma H, Borovkova N, et al. Characterisation of probiotic properties in human vaginal lactobacilli strains. Microb Ecol Health Dis 2016; 27: 30484.
25. Danielsen M, Wind A. Susceptibility of Lactobacillus spp. to antimicrobial agents. Int J Food Microbiol 2003; 82: 1-11.
26. Hoque MZ, Akter F, Hossain KM, Rahman MSM, Billah MM, Islam KMD. Isolation, identification and analysis of probiotic properties of Lactobacillus spp. from selective regional yoghurts. World J Dairy Food Sci 2010; 5: 39-46.
27. Devirgiliis C, Zinno P, Perozzi G. Update on antibiotic resistance in foodborne Lactobacillus and Lactococcus species. Front Microbiol 2013; 4: 301.
28. Rigo GV, Tasca T. Vaginitis: Review on drug resistance. Curr Drug Targets 2020; 21: 1672-1686.
29. Kiray E, Kariptas E, Azarkan SY. Evaluation of vaginal lactobacilli with potential probiotic properties and biotherapeutic effects isolated from healthy Turkish women. PSM Microbiol 2019; 4: 56-70.
30. Abriouel H, Casado Muñoz MDC, Lavilla Lerma L, Pérez Montoro B, Bockelmann W, Pichner R, et al. New insights in antibiotic resistance of Lactobacillus species from fermented foods. Food Res Int 2015; 78: 465-481.
31. Sirichoat A, Flórez AB, Vázquez L, Buppasiri P, Panya M, Lulitanond V, et al. Antibiotic resistance-susceptibility profiles of Enterococcus faecalis and Streptococcus spp. from the human vagina, and genome analysis of the genetic basis of intrinsic and acquired resistances. Front Microbiol 2020; 11: 1438.
32. Kassaa AI, Hamze M, Hober D, Chihib NE, Drider D. Identification of vaginal lactobacilli with potential probiotic properties isolated from women in North Lebonan. Microb Ecol 2014;67:722-734.
33. Hummel AS, Hertel C, Holzapfel WH, Franz CM. Antibiotic resistances of starter and probiotic strains of lactic acid bacteria. Appl Environ Microbiol 2007; 73: 730-739.
34. Sirichoat A, Flórez AB, Vázquez L, Buppasiri P, Panya M, Lulitanond V, et al. Antibiotic susceptibility profiles of lactic acid bacteria from the human vagina and genetic basis of acquired resistances. Int J Mol Sci 2020; 21: 2594.
35. Lim KT, Hanifah YA, Yusof M, Thong KL. ermA, ermC, tetM and tetK are essential for erythromycin and tetracycline resistance among methicillin-resistant Staphylococcus aureus strains isolated from a tertiary hospital in Malaysia. Indian J Med Microbiol 2012; 30: 203-207.
36. Begovic J, Huys G, Mayo B, D'Haene K, Florez AB, Lozo J, et al. Human vaginal Lactobacillus rhamnosus harbor mutation in 23S rRNA associated with erythromycin resistance. Res Microbiol 2009; 160: 421-426.
37. Goossens H. Assessing the safety of probiotics with regard to antibiotic resistance, in Functional dairy products. 2007, Elsevier. p. 427-442.
38. Delgado S, Flórez AB, Mayo B. Antibiotic susceptibility of Lactobacillus and Bifidobacterium species from the human gastrointestinal tract. Curr Microbiol 2005; 50: 202-207.
| Files | ||
| Issue | Vol 16 No 2 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v16i2.15356 | |
| Keywords | ||
| Antibiotic susceptibility; Iranian women; Probiotic; Resistance genes; Vaginal Lactobacillus | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Zare H, Izadi Amoli R, Zaboli F, Rezapour M, Kaboosi H. The phenotypic and genotypic antibiotic susceptibility of vaginal Lactobacillus with potential probiotic properties isolated from healthy women in northern Iran. Iran J Microbiol. 2024;16(2):227-235.
