Escherichia coli in betel leaves: prevalence, virulence characterization and antibiogram

Authors

DOI:

https://doi.org/10.3329/aajbb.v9i3.76177

Keywords:

E. coli, prevalence, virulence factors, antibiotic resistance, multidrug resistance, betel leaf contamination

Abstract

Escherichia coli is a significant foodborne pathogen, frequently linked to hemorrhagic diarrhea, especially in developing nations, where it presents considerable public health concerns. This study was conducted to examine the virulence gene profiles and antibiotic resistance patterns of E. coli strains isolated from piper betel leaves. A total of 100 betel leaf samples, including fresh (n = 60) and ready-to-eat (n = 40) specimens, were collected and tested for the presence of E. coli using standard diagnostic techniques, such as selective culture methods, staining, latex agglutination, and polymerase chain reaction (PCR) assays. Further, the identified E. coli isolates underwent PCR-based testing for virulence genes and disk diffusion assays to assess antibiotic susceptibility. Among the 100 samples screened, 4% (n = 4/100; 95% CI: 1.57–9.84; P = 0.1126) were identified as E. coli O157, and 33% (n = 33/100; 95% CI: 24.56–42.69; P = 0.4011) were classified as non-O157 isolates. The virulence gene stx1 was found in 10.81% of isolates, while stx2, eaeA, and hlyA genes were not detected in any samples. Antibiotic resistance analysis showed that all isolates (100%, 37/37) were resistant to amoxicillin and erythromycin, with 75.68% (28/37) demonstrating resistance to tetracycline. Notably, all isolates were fully susceptible to ceftriaxone and ciprofloxacin. A majority (72.97%, 27/37) of isolates were sensitive to streptomycin, and 67.57% (25/37) were sensitive to gentamicin. Additionally, 86.48% of the E. coli isolates exhibited multidrug resistance (MDR), showing 10 resistance patterns, including 8 MDR patterns. The most common MDR pattern was AMX-TE-E, observed in 56.76% (21/37) of isolates. One isolate demonstrated resistance to six of the eight tested antibiotics across four distinct classes, with the resistance pattern AMX-TE-GEN-S-E-AZM. The MAR indices for E. coli isolates ranged between 0.25 and 0.75. These findings highlight the significant threat posed to global public health by multidrug-resistant shiga toxin-producing E. coli found on piper betel leaves in urban environments.

Asian Australas. J. Biosci. Biotechnol. 2024, 9(3), 33-44

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References

Ababu A, D Endashaw and H Fesseha, 2020. Isolation and antimicrobial susceptibility profile of Escherichia coli O157: H7 from raw milk of dairy cattle in Holeta district, central Ethiopia. Int. J. Microbiol., 2020: 1-8.

Ali Almahdi AA and Y Kumar, 2019. Comparative study of antimicrobial activity of betel leaf extract and antibiotics against selected bacterial pathogens. Int. J. Curr. Microbiol. Appl. Sci., 8: 2009-2019.

Bauer AW, WMM Kirby, JC Sherris and M Turck, 1966. Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol., 45: 493-496.

Berger CN, SV Sodha, RK Shaw, PM Griffin, D Pink, P Hand and G Frankel, 2010. Fresh fruit and vegetables as vehicles for the transmission of human pathogens. Environ. Microbiol., 12: 2385-2397.

Beshiru A, AI Okoh and EO Igbinosa, 2022. Processed ready-to-eat (RTE) foods sold in Yenagoa Nigeria were colonized by diarrheagenic Escherichia coli which constitute a probable hazard to human health. PLoS ONE, 17: e0266059.

CLSI, 2018. Clinical and laboratory standards institute. Performance standards for antimicrobial susceptibility testing; 22nd informational supplement. CLSI Document M-100 S22. Wayne, Pennsylvania. Clinical and Laboratory Standards Institute, Wayne.

Fallah N, M Ghaemi, K Ghazvini, M Rad and A Jamshidi, 2021. Occurrence, pathotypes, and antimicrobial resistance profiles of diarrheagenic Escherichia coli strains in animal source food products from public markets in Mashhad, Iran. Food Control, 121: 107640.

Faruque MO, S Mahmud, MA Munayem, R Sultana, MT Molla, MF Ali, M Wasim, S Sarker and FZ Evamoni, 2019. Bacteriological analysis and public health impact of broiler meat: A study on Nalitabari Paurosova, Sherpur, Bangladesh. Adv. Microbiol., 9: 581-601.

Fazle R, Y Mahmuda, N Jamalun, N Ashikun, MC Fatema, O Yoshimitsu and RA Chowdhury, 2014. Bovine shiga toxin producing Escherichia coli O157:H7 of Bangladesh: Is it capable of causing diseases similar to clinical strains? African J. Microbiol. Res., 8: 147-154.

Gally DL and MP Stevens, 2017. Microbe profile: Escherichia coli O157: H7–notorious relative of the microbiologist’s workhorse. Microbiology, 163: 1-3.

Golberg D, Y Kroupitski, E Belausov, R Pinto and S Sela, 2011. Salmonella typhimurium internalization is variable in leafy vegetables and fresh herbs. Int. J. Food Microbiol., 145: 250-257.

Haileselassie M, H Taddele, K Adhana and S Kalayou, 2013. Food safety knowledge and practices of abattoir and butchery shops and the microbial profile of meat in Mekelle city, Ethiopia. Asian Paci. J. Trop. Med., 3: 407-412.

Haque, MM, MAR Sarker, RA Rifa, MA Islam and MM Khatun, 2017. Detection of food-borne bacteria in ready to eat betel leaf sold at local markets in Mymensingh. Vet. World, 10: 1040-1045.

Hassan J, MS Parvej, MB Rahman, MSR Khan, MT Rahman, T Kamal and KHMNH Nazir, 2014. Prevalence and characterization of Escherichia coli from rectal swab of apparently healthy cattle in Mymensingh, Bangladesh. Microbes and Health, 3: 12-14.

Heuvelink AE, K Wernars and E de Boer, 1996. Occurrence of Escherichia coli O157 and other verocytotoxin-producing E. coli in retail raw meats in the Netherlands. J. Food Prot., 59: 1267-1272.

Heuvelink AE, NCAJ van de Kar, JFGM Meis, LAH Monnens and WJG Melchers, 1995. Characterization of verocytotoxin-producing Escherichia coli O157 isolates from patients with haemolytic uraemic syndrome in Western Europe. Epidemiol. Infect., 115: 1-3.

Husna AA, MA Islam, MT Rahman and MM Khatun, 2015. Efficacy of vinegar, sorbitol and sodium benzoate in mitigation of Salmonella contamination in betel leaf. J. Adv. Vet. Anim. Res., 2: 190-194.

Islam A, M Rahman, A Nahar, A Khair and M Alam, 2015. Investigation of pathogenic Escherichia coli from diarrheic calves in selective area of Bangladesh. Bangladesh J. Vet. Med., 13: 45-51.

Islam M A, AS Mondol, E de Boer, RR Beumer, MH Zwietering, KA Talukder and AE Heuvelink, 2008. Prevalence and genetic characterization of shiga toxin-producing Escherichia coli isolates from slaughtered animals in Bangladesh. Appl. Environ. Microbiol., 74: 5414-5421.

Islam MA, AE Heuvelink, E de Boer, PD Sturm, RR Beumer, MH Zwietering, ASG Faruque, R Haque, DA Sack and KA Talukder, 2007. Shiga toxin-producing Escherichia coli isolated from patients with diarrhea in Bangladesh. J. Med. Microbiol., 56: 380-385.

Iswariya S and T Uma, 2017. Evaluation of in vitro anti-inflammatory and antimicrobial activity of aqueous and methanolic seed extracts of Citrullus lanatus. Int. J. Pharm. Sci., 9: 29.

Kabir SML, K Kikuchi, M Asakura, S Shiramaru, N Tsuruoka, A Goto, A Hinenoya and S Yamasaki, 2011. Evaluation of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the identification of Campylobacter strains isolated from diarrheal patients in Japan. Japanese J. Infect. Dis., 64: 19-27.

Kamal T, KHMNH Nazir, MS Parvej, MT Rahman, M Rahman, MFR Khan, WK Ansari, MM Ahamed, S Ahmed, ML Hossen, SN Panna and MB Rahman, 2018. Remedy of contamination of multidrug resistant Salmonella and Escherichia coli from betel leaves (Piper betle) keeping them fresh for long time. J. Adv. Vet. Anim. Res, 5: 73-80.

Karch H, PI Tarr and M Bielaszewska, 2005. Enterohaemorrhagic Escherichia coli in human medicine. Int. J. Med. Microbiol., 295: 405-418.

Mahfuza EJ, SM Sayem and MF Hassan, 2020. Trend of betel leaf production in Bangladesh: prospects and challenges. Res. Agri. Live. Fish., 7: 209-223.

Momtaz H, R Farzan, E Rahimi, FS Dehkordi and N Souod, 2012. Molecular characterization of shiga toxin-producing Escherichia coli isolated from ruminant and donkey raw milk samples and traditional dairy products in Iran. Sci. World J., 2012: 1-13.

Msolo L, BC Iweriebor and AI Okoh, 2020. Antimicrobial resistance profiles of diarrheagenic E. coli (DEC) and Salmonella species recovered from diarrheal patients in selected rural communities of the Amathole district municipality, eastern Cape province, South Africa. Infect. Drug Resist., 13: 4615-4626.

Munaomar M, K Hossain, F Afroz and M Salauddin, 2018. Antibiotic sensitivity pattern of bacteria isolated from infected betel leaf. IOSR J. Agri. Vet. Sci., 11: 82-91.

Nahar N, S Das, A Sufian, MK Islam and H Akter, 2018. Prevalence of microbial loads on betel leaf with special emphasis on multidrug resistance Salmonella spp and its public health implications. Int. J. Env. Agri. Biotechnol., 3: 1582-1589.

Okeke I, R Laxminarayan, ZA Bhutta, AG Duse, P Jenkins, TF O’Brien, A Pablos-Mendez and KP Klugman, 2005. Antimicrobial resistance in developing countries. Part I: recent trends and current status. Lancet Infect. Dis., 5: 481-493.

Osundiya OO, RO Oladele and OO Oduyebo, 2013. Multiple Antibiotic Resistance (MAR) indices of Pseudomonas and Klebsiella species isolates in Lagos University Teaching Hospital. African J. Clin. Exp. Microbiol., 14: 164-168.

Paton, AW and JC Paton, 1998. Detection and characterization of shiga toxigenic Escherichia coli by using multiplex PCR assays for stx1, stx2, eaeA, enterohemorrhagic E. coli hlyA, rfbO111and rfbO157. J. Clin. Microbiol., 36: 598-602.

Qadri F, AM Svennerholm, ASG Faruque and RB Sack, 2005. Enterotoxigenic Escherichia coli in developing countries: epidemiology, microbiology, clinical features, treatment, and prevention. Clin. Microbiol., 18: 465-483.

Ripon J, M Shahid, M Mahmud, S Das, M Rahman and KHMNH Nazir, 2021. Isolation and molecular characterization of shiga-toxin producing Escherichia coli from betel leaf (Piper betel L.). J. Adv. Vet. Anim. Res., 1: 12.

Sarker M, M Mannan, M Ali, M Bayzid, A Ahad and Z Bupasha, 2019. Antibiotic resistance of Escherichia coli isolated from broilers sold at live bird markets in Chattogram, Bangladesh. J. Adv. Vet. Anim. Res., 6: 272.

Sweeney MT, BV Lubbers, S Schwarz and JL Watts, 2018. Applying definitions for multidrug resistance, extensive drug resistance and pandrug resistance to clinically significant livestock and companion animal bacterial pathogens. J. Antimicrob. Chemother., 73: 1460-1463.

Talukdar PK, M Rahman, M Rahman, A Nabi, Z Islam, MM Hoque, HP Endtz and MA Islam, 2013. Antimicrobial resistance, virulence factors and genetic diversity of Escherichia coli isolates from household water supply in Dhaka, Bangladesh. PLoS One, 8: e61090.

Uddin J, K Hossain, S Hossain, K Saha, FT Jubyda, R Haque, B Billah, AA Talukder, AK Parvez and SK Dey, 2019. Bacteriological assessments of foodborne pathogens in poultry meat at different super shops in Dhaka, Bangladesh. Ital. J. Food Saf., 8: 6720.

Wang RF, WW Cao and CE Cerniglia, 1996. PCR detection and quantitation of predominant anaerobic bacteria in human and animal fecal samples. Appl. Environ. Microbiol., 62: 1242-1247.

Wieler LH, M Tigges, F Ebel, S Schäferkordt, S Djafari, T Schlapp, G Baljer and T Chakraborty, 1996. The enterohemolysin phenotype of bovine shiga-like toxin-producing Escherichia coli (SLTEC) is encoded by the EHEC-hemolysin gene. Vet. Microbiol., 52: 153-164.

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Published

2024-11-18

How to Cite

Islam, N., Islam, M. R., Liza, R. A., Nobi, M. A., Islam, S. S., Arif, M., Khan, M. F. R., Samosornsuk, S., Samosornsuk, W., & Kabir, S. L. (2024). Escherichia coli in betel leaves: prevalence, virulence characterization and antibiogram. Asian-Australasian Journal of Bioscience and Biotechnology, 9(3), 33–44. https://doi.org/10.3329/aajbb.v9i3.76177

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Research Articles