Detection of antibiotic resistance genes in bacterial isolates from most touched surfaces of public transports in Sagamu, Ogun state, Nigeria

Abstract

Background and Aims: The role of fomites in the transmission of infectious diseases is becoming more important because of the possibility that contaminated surfaces act as reservoirs of diseases. The aim of this study was to ascertain the level of bacterial contamination and the prevalence of antibiotic-resistant genes on the most touched surfaces on public transport. Methods: One hundred samples were collected from door handles and armrests of buses and tricycles, respectively at Isale-Oko motor garage, Sagamu. Bacteria were isolated from the samples and identified following standard microbiological techniques. Antibiotic susceptibility testing was done with the Kirby-Bauer disc diffusion method. The presence of antibiotic-resistant genes vanA, drfA, dfrG genes, and extended-spectrum betta-lactamase (ESBL) genes in Gram-negatives were screened by polymerase chain reaction (PCR) method. Results: Out of the samples tested, 91% were positive for bacterial contamination. Among the 91 positive samples, 126 bacteria were identified, comprising 98 Gram-positive and 28 Gram-negative bacterial isolates. Staphylococcus aureus had the highest overall frequency of occurrence with 62 (49.2%) isolates. Among the Gram-positives, azithromycin resistance was present in 35(56.5%) S. aureus and 19(52.8%) Staphylococcus epidermidis. Salmonella species was the most resistant to ciprofloxacin (100%). dfrG was the most detected among trimethoprim-resistant genes occurring in 11(55%) of multidrug-resistant S. aureus and 6(54.6%) of S. epidermidis. vanA gene was present in S. aureus (20%). dfrA was present in only Klebsiella pneumoniae and Esherichia coli. E. coli and Shigella species carried 𝑏𝑙𝑎𝑇𝐸𝑀 while 𝑏𝑙𝑎𝑆𝐻𝑉 was found in Pseudomonas aeruginosa. Conclusion: The most-touched surfaces of public transportation can serve as a substantial source of spread for potentially harmful bacteria.

Keywords

• Antibiotic resistance
• Extended-spectrum beta-lactamase
• Bacterial isolates
• Public transportation

References

1. Allcock, S., Young, E.H., Holmes, M., Gurdasani, D., Dougan, G., Sandhu, M.S., . . . Török, M.E. (2017). Antimicrobial re-sistance in human populations: challenges and opportunities. Global Health Epidemiology and Genomics, 10(2): Article e4. http://dx.doi.org/10.1017/gheg.2017.4 google scholar
2. Argudın, M.A., Tenhagen, B.A., Fetsch, A., Sachsenro''der, J., Ka''sbohrer, A., Schroeter, A., ... Guerra, B. (2011). Virulence and resistance determinants of German Staphy-lococcus aureus ST398 isolates from nonhuman sources. Applied Environmental Microbiology, 77, 3052-3060. http://dx.doi.org/10.1128/AEM.02260-10. google scholar
3. Ashgar, S.S. &El-Said, H.M. (2012). Pathogenic bacteria asso-ciated with different public environmental sites in Mecca City. Open Journal of Medical Microbiology, 2, 133-137. http://dx.doi.org/10.4236/ojmm.2012.24020 google scholar
4. Atata, R.F., Ibrahim, Y.K.E., Olurinola, P.F., Giwa, A., Akanbi, A.A. & Sani, A.A. (2010). Clinical bacterial isolates from a hospi-tal environment as agents of surgical wound nosocomial infec-tions. Journal of Pharmacy and Bioresources, 7(2), 146-155 http://dx.doi.org/10.4314/jpb.v7i2.15 google scholar
5. Bauer, A.W., Kirby, W.M., Sherris, J.C. & Turck, M. (1966). Antibi-otic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology,45(4), 493-496. google scholar
6. Bhatta, D.R., Hamal, D., Shrestha, R. Hosuru Subramanya, S., Baral, N., Singh, R.K., . . . Gokhale, S. (2018). Bacterial contamina-tion of frequently touched objects in a tertiary care hospital of Pokhara, Nepal: how safe are our hands? Antimicrobial Resistance and Infection Contro,. 7, 97. http://dx.doi.org/10.1186/s13756-018-0385-2 google scholar
7. Birteksöz Tan, A.S. & Erdoğdu G. (2017). Microbiological burden of public transport vehicles. Istanbul Journal of Pharmacy, 47(2), 52-56. http://dx.doi.org/10.5152/IstanbulJPharm.2017.008 google scholar
8. Cave, R., Cole, J. & Mkrtchyan, H.V. (2021). Surveillance and preva-lence of antimicrobial resistant bacteria from public settings within urban built environments: Challenges and opportunities for hygiene and infection control. Environment International, 157: Article 106836. http://dx.doi.org/10.1016/j.envint.2021.106836. google scholar

9. Cheesbrough, M. (2006). District Laboratory Practice in Tropical Countries. Part 2, 2nd Edition, Cambridge University Press Pub-lication, South Africa. google scholar
10. Chowdhury, T., Mahmud, A., Barua, A., Khalil, M.D.I., Chowd-hury, R., Ahamed, F. & Dhar, K. (2016). Bacterial contamina-tion on hand touch surfaces of public buses in Chittagong city, Bangladesh. Journal of Environmental Science, Toxicology and Food Technology, 10(4), 48-55. http://dx.doi.org/ 10.9790/2402-1004034855 google scholar
11. Clinical and Laboratory Standards Institute (CLSI), (2022). Perfor-mance Standards for Antimicrobial Susceptibility Testing, 32nd Edition. google scholar
12. Coelho, C., de Lencastre, H. & Aires-de-Sousa, M. (2017). Frequent occurrence of trimethoprim-sulfamethoxazole hetero-resistant Staphylococcus aureus isolates in different African countries. Eu-ropean Journal of Clinical Microbiology and Infectious Diseases, 36, 1243-1252. http://dx.doi.org/ 10.1007/s10096-017-2915-x google scholar
13. Dommguez, M., Miranda, C.D., Fuentes, O., de la Fuente, M., Godoy, F.A., Bello-Toledo, H & Gonzalez-Rocha, G. (2019). Occurrence of Transferable Integrons and sul and dfr Genes among Sulfonamide-and/or Trimethoprim-Resistant Bacteria Iso-lated from Chilean Salmonid Farms. Frontiers in Microbiology, 10, 748. http://dx.doi.org/ 10.3389/fmicb.2019.00748 google scholar
14. Dutka-Malen, S., Evers, S. & Courvalin, P. (1995). Detec-tion of glycopeptide resistance genotypes and identifica-tion to the species level of clinically relevant enterococci by PCR. Journal of Clinical Microbiology, 33(1), 24-27. http://dx.doi.org/10.1128/jcm.33.1.24-27.1995 google scholar
15. Edelstein, M., Pimkin, M., Edelstein. I. & Stratchounski, L. (2003). Prevalence and molecular epidemiology of CTX-M extended-spectrum p—lactamase -producing Escherichia coli and Kleb-siella pneumoniae in Russian hospitals. Antimicrobial Agents Chemotherapy, 47: 3724-3732. google scholar
16. Fernandes, C.J., Fernandes, L.A. & Collignon, P. (2005). Cefoxitin resistance as a surrogate marker for the de-tection of methicillin-resistant Staphylococcus aureus. Journal of Antimicrobial Chemotherapy, 55(4), 506-510. http://dx.doi.org/10.1093/jac/dki052 google scholar
17. Furuya, H. (2007). Risk of transmission of airborne in-fection during train commute based on a mathematical model. Environmental Health Preventive Medicine, 12, 78-83. http://dx.doi.org/10.1007/BF02898153 google scholar
18. Grape, M., Motakefi, A., Pavuluri, S. & Kahlmeter, G. (2007). Standard and real-time multiplex PCR methods for detection of trimethoprim resistance dfr genes in large collections of bacteria. Clinical Microbiology and Infection, 13, 1112-1118. http://dx.doi.org/ 10.1111/j.1469-0691.2007.01807.x. google scholar
19. Guimera, R., Mossa, S., Turtschi, A. & Amara, L.A.N. (2005). The worldwide air transportation network: anomalous centrality, com-munity structure, and cities’ global roles. Proceedings of the Na-tional Academy of Sciences of the United States of America, 102, 7794-7799. http://dx.doi.org/10.1073/pnas.0407994102 google scholar
20. Kahsay, A.G., Asgedom, S.W. & Weldetinsaa, H.L. (2019). En-teric bacteria, methicillin-resistant S. aureus and antimicro-bial susceptibility patterns from buses surfaces in Mekelle city, Tigray, Ethiopia. BMC Research Notes, 12, 337. http://dx.doi.org/10.1186/s13104-019-4366-4371 google scholar
21. Magiorakos, A.-P., Srinivasan, A., Carey, R.B., Carmeli, Y., Falagas, M.E.C.G., Giske, C.G., . . . Monnet, D.L. (2012). Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clinical Microbiology and Infection, 18(3), 268-281. http://dx.doi.org/10.1111/j.1469-0691.2011.03570.x. google scholar
22. Maynard, C., Fairbrother, J.M., Bekal, S., Sanschagrin, F., Levesque, R.C., Brousseau, R., . . . Harel, J. (2003). An-timicrobial resistance genes in enterotoxigenic Escherichia coli O149:K91 isolates obtained over a 23-year period from pigs. Antimicrobial Agents Chemotherapy, 47(10), 3214-3221. http://dx.doi.org/10.1128/AAC.47.10.3214-3221.2003 google scholar
23. Montravers, P. & Eckmann, C. (2021). Cotrimoxazole and clindamycin in skin and soft tissue infections. Cur-rent Opinion in Infectious Diseases, 34(2), 63-71. http://dx.doi.org/10.1097/QCO.0000000000000698 google scholar
24. Muley, D., Shahin, M., Dias C. & Abdullah, M. (2020). Role of Transport during Outbreak of Infectious Dis-eases: Evidence from the Past. Sustainability, 12, 7367. http://dx.doi.org/https://doi.org/10.3390/su12187367 google scholar
25. Nurjadi, D., Olalekan, A.O., Layer, F., Shittu, A.O., Alabi, A., Ghe-bremedhin B., ... Zanger, P. (2014). Emergence of trimethoprim resistance gene dfrG in Staphylococcus aureus causing human in-fection and colonization in sub-Saharan Africa and its import to Europe. Journal of Antimicrobial Chemotherapy, 69, 2361-2368. http://dx.doi.org/10.1093/jac/dku174. google scholar
26. Nwankwo, E.O., Okey-kalu, E.U. & Eze F.A. (2023). Bac-terial Contamination of Door Handles of Commer-cial Buses in Umuahia Metropolis Abia State. Suan Sunandha Science and Technology Journal, 10(1), 54-61. http://dx.doi.org/https://doi.org/10.53848/ssstj.v10i1.414 google scholar
27. Olaniran, O.B., Adeleke, O.E., Donia, A., Shahid, R. & Bokhari, H. (2021). Incidence and Molecular Characterization of Carbapen-emase Genes in Association with Multidrug-Resistant Clinical Isolates of Pseudomonas aeruginosa from Tertiary Healthcare Facilities in Southwest Nigeria. Current Microbiology, 79(1), 27. http://dx.doi.org/10.1007/s00284-021-02706-3 google scholar
28. Otter, J.A. & French, G.L. (2009). Bacterial contamination on touch surfaces in the public transport system and in public areas of a hospital in London. Letters in Applied Microbiology, 49(6), 803807. http://dx.doi.org/10.1111/j.1472-765X.2009.02728.x google scholar
29. Reeve, S.M., Scocchera, E.W., G-Dayanadan, N., Keshipeddy, S., Krucinska, J., Hajian, B., Ferreira, J., Nailor, M., Aeschlimann, J., Wright, D.L......Anderson, A.C. (2016). google scholar
30. MRSA isolates from United States hospitals carry dfrG and dfrK resistance genes and succumb to propargyl-linked antifolates. Cell Chemical Biology, 23(12), 1458-1467. http://dx.doi.org/10.1016/j.chembiol.2016.11.007. google scholar
31. Rodrigue, J-P, (2020) The Geography of Transport Systems, Fifth Edi-tion, London: Routledge. 456 pages. ISBN: 978-0-367-36463-2. https://doi.org/10.4324/9780429346323.‘ google scholar
32. Rossato, A.M., Primon-Barros, M., Rocha, L.D.L., Reiter, K.C., Dias, C.A.G. & d’Azevedo, P.A. (2020). Resistance profile to antimicro-bial agents in methicillin-resistant Staphylococcus aureus isolated from hospitals in South Brazil between 2014-2019. Revista da So-ciedade Brasileira de Medicina Tropical, 53, Article e20200431. http://dx.doi.org/10.1590/0037-8682-0431-2020. google scholar
33. Rusin, P., Maxwell, S. & Gerba, C. (2002). Comparative surface-to-hand and fingertip-to-mouth transfer efficiency of gram-positive bacteria, gram-negative bacteria and phage. Journal of Applied Microbiology, 93, 585-592. http://dx.doi.Org/10.1046/j.1365— 2672.2002.01734.x google scholar
34. Shittu, A.O., Okon, K., Adesida, S., Oyedara, O., Witte, W., Strom-menger, B., . . . Nübel, U. (2011). Antibiotic resistance and molec-ular epidemiology of Staphylococcus aureus in Nigeria. BMC Mi-crobiology, 11, 92. http://dx.doi.org/ 10.1186/1471-2180-11-92 google scholar
35. Tatem, A.J., Rogers, D.J. & Hay, S.I. (2006). Global Transport Net-works and Infectious Disease Spread. Advances in Parasitology, 62, 293-343. http://dx.doi.org/ 10.1016/S0065-308X(05)62009-X google scholar
36. World Health Organization (WHO) 2021. Antimicrobial resistance. Downloaded on March 8, 2023. https://www.who.int/news-room/fact-sheets/detail/antimicrobial-resistance google scholar
37. World Health Organization, (2020). Updated WHO recommendations for international traffic in relation to COVID-19 outbreak. Downloaded on March 8, 2023. https://www.who.int/news-room/articles-detail/updated-who-recommendations-for-international-traffic-in-relation-to-covid-19-outbreak google scholar
38. Yeh, P.J., Simon, D.M., Millar, J.A., Alexander, H.F. & Franklin, D. (2011). A diversity of Antibiotic-resistant Staphylococcus spp. in a Public Transportation System. Public Health and Research Perspectives, 2(3), 202-209. http://dx.doi.org/10.1016/j.phrp.2011.11.047 google scholar
39. Yu, H.S., Lee, J.C., Kang, H.Y. & Yeong, Y.S. (2004). Prevalence of dfr genes associated with integrons and dissemination of dfrA17 among urinary isolates of Escherichia coli in Korea. Journal of Antimicrobial Chemotherapy, 53(3), 445-450. http://dx.doi.org/ 10.1093/jac/dkh097 google scholar