Plasmid profiles of antibiotic-resistant opportunistic strains of the Enterobacteriaceae family in the microbiota of men with urogenital tract inflammatory pathology

T. Sklyar; N. Kurahina; T. Lykholat; O. Lykholat; M. Papiashvili

doi:10.15421/0225096

T. Sklyar Oles Honchar Dnipro National University
N. Kurahina Oles Honchar Dnipro National University
T. Lykholat Oles Honchar Dnipro National University
O. Lykholat University of Customs and Finance
M. Papiashvili HCA Healthcare MidAmerica Division

Keywords: enterobacteria, Escherichia coli, Proteus mirabilis, Klebsiella oxytoca, plasmids, antibiotic resistance, multidrug-resistant strains, electrophoregram.

Abstract

Bacterial resistance to antimicrobial drugs is a serious public health problem. Plasmids represent one of the most diff i cult problems in combating the spread of antibiotic resistance. The article presents the results of a real-time PCR study of the microbiota of 257 men with inflammatory processes of the urogenital tract. The role of representatives of the opportunistic microbiota in the occurrence of infectious and inflammatory processes of the urogenital system in men was established. The composition of the microbiota was represented by bacteria of the Enterobacteriaceae family in 39.3% of cases, Enter o coccus spp. – 10.9%, Haemophilus spp. – 3.1% and Pseudomonas aeruginosa – in 0.4% of cases. Analysis of the antibiotic sensitivity spectra of the bacterial intestinal group allowed us to establish that among the isolated enterobacteria monoresi s tance to ampicillin (21 – 23%) and resistance to ampicillin and amoxicillin/clavulanate (11 – 14%) were the most common. The work also revealed multiple resistance of clinical strains of enterobacteria to antibiotics: 8% of strains were found to be resistant to cephalosporins of the 3rd and 4th generations and ampicillin, i.e. to β-lactams (phenotype ApCpmCtaCrdCtx); 5% of strains were resistant to 7 and 9 drugs. For Escherichia , Klebsiella and Proteus (resistance to Ap and Ap Am/Cl) only two spectra were characteristic. At the same time, seven spectra were common to Es c herichia coli and Klebsiella sp., five – to E. coli and Proteus sp., three – to Proteus sp. and Klebsiella sp. Plasmids of different sizes were isolated from clinical polyresistant strains of E. coli , Proteus mirabilis , Klebsiella oxytoca . However, some multiresistant strains didn’t contain plasmids. Direct relationship between antibiotic resistance and the plasmid profile of E. coli wasn’t found. In experimental clinical strains of enterobacteria the presence of such plasmid spectrum opens up prospects for studying the widespread distribution of multidrug-resistant strains.

References

Akter, S., Chwdhury, M. A., & Mina, S. A. (2021). Antibiotic resistance and plasmid profiling of Escherichia coli isolated from human sewage samples. Microbiology Insights, 14, 1–6.

Alekshun, M. N., & Levy S. B. (2007). Molecular mechanisms of antibacterial multidrug resistance. Cell, 128 (6), 1037–1050.

Alonso-Del Valle, A., Leon-Sampedro, R., Rodriguez-Beltran, J., DelaFuente, J., Hernandez-Garcia, M., Ruiz-Garbajosa, P., Canton, R., Pena-Miller, R., & San Millan, A. (2021). Variability of plasmid fitness effects contributes to plasmid persistence in bacterial communities. Nature Communication, 12(1), 2653.

Aminul, P., Anwar, S., Molla, M. & Miah, R. (2021). Evaluation of antibiotic resistance patterns in clinical isolates of Klebsiella pneumoniae in Bangladesh. Biosafety and Health, 3(6), 301–306.

Benz, F., & Hall, A. R. (2023). Host-specific plasmid evolution explains the variable spread of clinical antibiotic-resistance plasmids. Proceedings of the National Academy of Science of the United States of America, 120(15), e2212147120.

Bethke, J. H., Davidovich, A., Cheng, L., Lopatkin, A. J., Song, W., Thaden, J. T., Fowler, V. G., Xiao, M., & You, L. (2020). Environmental and genetic determinants of plasmid mobility in pathogenic Escherichia coli. Science Advance, 6(4), eaax3173.

Bevan, E., Jones, A., & Hawkey, P. (2017). Global epidemiology of CTX-M β-lactamases: Temporal and geographical shifts in genotype, Journal of Antimicrobial Chemotherapy, 72, 2145–2155.

Burova, L., Korneichuk, E., Kushkina, A., & Tovkach, F. (2012). Plasmid profile, colicinogeny and phage sensitivity as indicators of the dynamics of Escherichia coli populations in the human gut. Microbiological Journal, 74(5), 99–107.

Bush, K., & Bradford, P. A. (2020). Epidemiology of β-lactamase-producing pathogens. Clinical Microbiology Reviews, 33(2), e00047-19.

Cairns, J., Ruokolainen, L., Hultman, J., Tamminen, M., Virta, M., & Hiltunen, T. (2018). Ecology determines how low antibiotic concentration impacts community composition and horizontal transfer of resistance genes. Communications Biology, 35, 21–29.

CLSI (2018). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. 11th ed. CLSI standart M07. Wayne: Clinical and Laboratory Standarts Institute. 13.

CLSI (2020). Performance standards for antimicrobial susceptibility testing. 30th ed. CLSI supplement M100. Wayne: Clinical and Laboratory Standards Institute. 332.

David, S., Cohen, V., Reuter, S., Sheppard, A. E., Giani, T., Parkhill, J., Rossolini, M. G., Feil, E. J., Grundmann, H., & Aanensen, D. M. (2020). Integrated chromosomal and plasmid sequence analyses reveal diverse modes of carbapenemase gene spread among Klebsiella pneumoniae. Proceedings of the National Academy of Science, 117(40), 25043–25054.

David, S., Reuter, S., Harris, S. R., Glasner, C., Feltwell, T., Argimon, S., Abudahab, K., Goater, R., Giani, T., Errico, G., Aspbury, M., Sjunnebo, S., Feil, E. J., Rossolini, G. M., Aanensen, D. M., & Grundmann, H. (2019). Epidemic of carbapenem-resistant Klebsiella pneumoniae in Europe is driven by nosocomial spread. Nature Microbiology, 4(11), 1919–1929.

Delaney, S., Murphy, R., & Walsh, F. A. (2018). A comparison of methods for the extraction of plasmids capable of conferring antibiotic resistance in a human pathogen from complex broiler cecal samples. Frontiers in Microbiology, 9, 1731.

Fernandez-Calvet, A., Toribio-Celestino, L., Alonso-Del Valle, A., Sastre-Dominguez, J., Valdes-Chiara, P., San Millan, A., & DelaFuente, J. (2023). The distribution of fitness effects of plasmid pOXA-48 in clinical enterobacteria. Microbiology, 169, 001369.

Hall, J. P., Wood, A. J., Harrison, E., & Brockhurst, M. A. (2016). Source-sink plasmid transfer dynamics maintain gene mobility in soil bacterial communities. Proceedings of the National Academy of Science, 113(29), 8260–8265.

Husna, A., Rahman, M., Badruzzaman, A., Sikder, M., Islam, M., Rahman, M., Alam, J., & Ashour, H. (2023). Extended-spectrum β-lactamases (ESBL): Challenges and opportunities. Biomedicines, 11(11), 2937–2941.

Jia, Y., Lu, H., & Zhu, L. (2022). Molecular mechanism of antibiotic resistance induced by mono- and twin-chained quaternary ammonium compounds. Science of the Total Environment, 832, 155090.

Johnson, T., & Nolan, L. K. (2009). Pathogenomics of the virulence plasmids of Escherichia coli. Microbiology and Molecular Biology Reviews, 73(4), 750–774.

Kado, C., & Liu, S. (1981). Rapid procedure for detection and isolation of large and small plasmids. Journal of Bacteriology, 145(3), 1365–1373.

Keyi, Y., Zhenzhou, H., Yue, X., He, G., Xuemei, B., & Duochun, W. (2024). Global spread characteristics of CTX-M-type extended-spectrum β-lactamases: A genomic epidemiology analysis. Drug Resistance Updates, 73, 101036.

Kosterlitz, O., Grassi, N., Werner, B., McGee, R. S., Top, E. M., & Kerr, B. (2023). Evolutionary "crowdsourcing": Alignment of fitness landscapes allows for cross-species adaptation of a horizontally transferred gene. Molecular Biology and Evolution, 40(11), msad237.

Larsson, D., & Flach, C. (2022). Antibiotic resistance in the environment. Nature Reviews Microbiology, 20(5), 257–269.

Li, L., Dechesne, A., Madsen, J. S., Nesme, J., Sorensen, S. J., & Smets, B. F. (2020). Plasmids persist in a microbial community by providing fitness benefit to multiple phylotypes. The ISME Journal, 14(5), 1170–1181.

Madrazo, M., Esparcia, A., López-Cruz, I., Alberola, J., Piles, L., Viana, A., Eiros, J., & Artero, A. (2021). Clinical impact of multidrug-resistant bacteria in older hospitalized patients with community-acquired urinary tract infection. BMC Infectious Diseases, 21(1), 1232–1235.

Meng, M., Li, Y., & Yao, H. (2022). Plasmid-mediated transfer of antibiotic resistance genes in soil. Antibiotics, 11(4), 525.

Mohapatra, D., Debata, N., & Singh, S. (2018). Extensively drug-resistant and pandrug-resistant Gram-negative bacteria in a tertiary-care hospital in Eastern India: A 4-year retrospective study. Journal of Global Antimicrobial Resistance, 15, 246–249.

Paitan, Y. (2018). Current trends in antimicrobial resistance of Escherichia coli. Current Topics in Microbiology and Immunology, 416, 181–211.

Palkovicova, J., Sukkar, I., Delafuente, J., Valcek, A., Medvecky, M., Jamborova, I., Bitar, I., Phan, M.-D., San Millan, A., & Dolejska, M. (2022). Fitness effects of bla CTX-M-15-harbouring F2:A1: B-plasmids on their native Escherichia coli ST131 H 30Rx hosts. Journal of Antimicrobial Chemotherapy, 77(11), 2960–2963.

Pusparajah, P., Letchumanan,V., Goh, B., & McGaw, L. (2022). Editorial: Novel approaches to the treatment of multidrug-resistant bacteria. Frontiers in Pharmacology, 13, 972935.

San Millan, A. (2018). Evolution of plasmid-mediated antibiotic resistance in the clinical context. Trends in Microbiology, 26(12), 978–985.

San Millan, A., & MacLean, R. C. (2017). Fitness costs of plasmids: A limit to plasmid transmission. Microbiology Spectrum, 5(5), 16.

Saravanan, M., Ramachandran, B., & Barabadi, H. (2018). The prevalence and drug resistance pattern of extended spectrum β-lactamases (ESBLs) producing Enterobacteriaceae in Africa. Microbial Pathogenesis, 114, 180–192.

Sklyar, T., Gavryliuk, V., Lavrentievа, K., Kurahina, N., Lykholat, T., Zaichenko, K., Papiashvili, M., Lykholat, O., & Stepansky, D. (2021). Monitoring of distribution of antibiotic-resistant strains of microorganisms in patients with dysbiosis of the urogenital tract. Regulatory Mechanisms in Biosystems, 12(2), 199–205.

Sklyar, T., Kurahina, N., Lavrentieva, K., Burlaka, V., & Lykholat, T. (2021). Autonomic (mobile) genetic elements of bacteria and their hierarchy. Cytology and Genetics, 55(3), 256–269.

Sklyar, T., Lavrentievа, K., Kurahina, N., Lykholat, T., Papiashvili, M., Lykholat, O., & Stepanskyi, D. (2022). Monitoring of enterobacteria strains with producing of β-lactamases in males with infectious-inflammatory diseases of urogenital tract. Medychni Perspektyvy, 27(2), 110–118.

Tischendorf, J., de Avila, R. A., & Safdar, N. (2016). Risk of infection following colonization with carbapenem-resistant Enterobactericeae: A systematic review. American Journal of Infection Control, 44(5), 539–543.

Zhu, X., Tang, Q., Zhou, X., & Momeni, M. (2024). Antibiotic resistance and nanotechnology: A narrative review. Microbial Pathogenesis, 193, 106741.