Bacteriological and Molecular Characterization of Diarrheagenic Escherichia Coli Pathotypes From Children in Duhok City, Iraq
DOI:
https://doi.org/10.25271/sjuoz.2020.8.1.702Keywords:
Diarrheagenic Escherichia coli, Stool, MacConkey-Sorbitol agar, Virulence factors, PCRAbstract
Background: A variety of diarrheagenic E coli (DEC) are responsible for causing different types of diarrhea in children especially in developing courtiers.
Objectives: This study was primarily aimed to isolate and bacteriological characterizing of E coli from diarrheic infant stool and to investigate their antibiotic resistance patterns and then using molecular identification of DEC pathotypes for better discrimination.
Methods: Total of 400 fresh stools specimens were collected from children with diarrhea in Heevi Hospital in Duhok city, Iraq. The samples were cultured on selective media such as (MacConkey and MacConkey sorbitol agar). Colonies were identified through biochemical reaction and VITEK 2 system and then antibiotic susceptibility profiles were determined.
Results: A total of 349(87.2%) samples were yielded positive for growth of E coli. Out of these, 50 phenotypically-identified E coli were then subjected to PCR assay targeting certain virulence factors (alt, eae, sxt1 and sxt2) for discrimination of pathotyes. 13/50(26%) Enterotoxigenix E. coli (ETEC) was detected, 5/50(10%) Enterohemorrhagic E coli EHEC was detected, while no Enteropathogenic E coli (EPEC) was detected. All pathotypes were more frequent in samples from male children aged between 2-3 years that were artificial feeding pattern. Moreover, all pathotypes expressed high resistant to ampicillin, cephalosporin and tetracycline while little resistance to imipenem was observed. Conclusion: The study concludes presence of diarrheagenic E coli pathotypes in our community causing diarrhea in children and emphasize on using of PCR assay for best discrimination.
References
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Spano, L.C., da Cunha, K.F., Monfardini, M.V., Bergamaschi, R.D., Isabel, F., Affonso, C. (2017). High prevalence of diarrheagenic Escherichia coli carrying toxin-encoding genes isolated from children and adults in southeastern Brazil. BMC Infect Dis 17, 773 (2017).
Sperandio, V., & Pacheco, A. R. (2012). Shiga toxin in enterohemorrhagic E. coli: regulation and novel anti-virulence strategies. Frontiers in cellular and infection microbiology, 2, 81
Sudershan, R., Kumar, R. N., Kulkarni, B., Kashinath, L., Bhaskar, V., & Polasa, K. (2014). E coli pathotypes and their antibiotic resistance in young children with diarrhea in Hyderabad, India. Int Journal Curr Microbiol App Sci, 3(9), 647-654.
Toma, C., Lu, Y., Higa, N., Nakasone, N., Chinen, I., Baschkier, A., Iwanaga, M. (2003). Multiplex PCR assay for identification of human diarrheagenic Escherichia coli. Journal of clinical microbiology, 41(6), 2669-2671.
Versalovic, J., Carroll, K., Funke, G., Jorgensen, J., Landry, M., Warnock, D. (2011). Manual of clinical microbiology, ASM Press. Washington, DC.
Weinstein, M., Patel, J., Bobenchik, A. (2017). Clinical and laboratory standards institute. Performance Standards for Antimicrobial Susceptibility Testing, 27th ed.; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 296.
Arif, S. K., & Salih, L. I. (2010). Identification of Different Categories of Diarrheagenic" Escherichia coli" in Stool Samples by Using Multiplex PCR Technique. Asian Journal of Medical Sciences, 2(5), 237-243.
Bauer, A. (1966). Antibiotic Susceptibility Testing by a standardized single disc method. Am J clin pathol, 45, 149-158.
Black, R. E., Cousens, S., Johnson, H. L., Lawn, J. E., Rudan, I., Bassani, D. G., Cibulskis, R. (2010). Global, regional, and national causes of child mortality in 2008: a systematic analysis. The lancet, 375 , 1969-1987.
Canizalez., R. A., Flores-Villaseñor., H.M., Gonzalez., N. E., Velazquez., R. J, Vidal., J.E., Muro- Amador., S, et al. (2016) Surveillance of Diarrheagenic Escherichia coli Strains Isolated from Diarrhea Cases from Children, Adults and Elderly at Northwest of Mexico. Front. Microbiol. 7, 1924.
Cheesbrough, M. (2006). District laboratory practice in tropical countries: Cambridge university press.
El Metwally, H. A., Ibrahim, H. A., El-Athamna, M. N., & Amer, M. A. (2007). Multiplex PCR for detection of diarrheagenic Escherichia coli in Egyptian children. Journal of Medical Sciences, 7(2), 255-262.
Engelkirk, P. G., & Duben-Engelkirk, J. L. (2008). Laboratory diagnosis of infectious diseases: essentials of diagnostic microbiology: Lippincott Williams & Wilkins.
Fujioka, M., Otomo, Y., & Ahsan, C. R. (2013). A novel single-step multiplex polymerase chain reaction assay for the detection of diarrheagenic Escherichia coli. Journal of microbiological methods, 92(3), 289-292.
Haghi, F., Zeighami, H., Hajiahmadi, F., Khoshvaght, H., & Bayat, M. (2014). Frequency and antimicrobial resistance of diarrhoeagenic Escherichia coli from young children in Iran. Journal of medical microbiology, 63(3), 427-432.
Hellard, M. E., Sinclair, M. I., Hogg, G. G., & Fairley, C. K. (2000). Prevalence of enteric pathogens among community based asymptomatic individuals. Journal of gastroenterology and hepatology, 15(3), 290-293.
Ifeanyi, C. I. C., Ikeneche, N. F., Bassey, B. E., Al-Gallas, N., Aissa, R. B., & Boudabous, A. (2015). Diarrheagenic Escherichia coli pathotypes isolated from children with diarrhea in the Federal Capital Territory Abuja, Nigeria. The Journal of Infection in Developing Countries, 9(02), 165-174.
Kaper, J. B., Nataro, J. P., Mobley, H. L. (2004). Pathogenic Escherichia coli. Nature reviews microbiology, 2(2), 123-140.
Khalil, Z. K. (2015). Isolation and identification of different diarrheagenic (DEC) Escherichia coli pathotypes from children under five years old in Baghdad. Iraqi journal of community medicine, 28(3), 126-132.
Liu, D. (2011). Molecular detection of human bacterial pathogens: 1st edition, CRC press.
McAuliffe, G. N., Anderson, T. P., Stevens, M., Adams, J., Coleman, R., Mahagamasekera, P., Jennings, L. C. (2013). Systematic application of multiplex PCR enhances the detection of bacteria, parasites, and viruses in stool samples. Journal of Infection, 67(2), 122-129.
Nessa, K., Ahmed, D., Islam, J., Kabir, F. L., & Hossain, M. A. (2007). Usefulness of a multiplex PCR for detection of diarrheagenic Escherichia coli in a diagnostic microbiology laboratory setting. Bangladesh Journal of Medical Microbiology, 1(2), 38-42.
Nguyen, T. V., Le Van, P., Le Huy, C., Gia, K. N., Weintraub, A. (2005). Detection and characterization of diarrheagenic Escherichia coli from young children in Hanoi, Vietnam. Journal of clinical microbiology, 43(2), 755-760.
Presterl, E., Zwick, R. H., Reichmann, S., Aichelburg, A., Winkler, S., Kremsner, P. G., & Graninger, W. (2003). Frequency and virulence properties of diarrheagenic Escherichia coli in children with diarrhea in Gabon. The American journal of tropical medicine and hygiene, 69 (4), 406-410.
Rahouma, A., Klena, J. D., Krema, Z., Abobker, A. A., Treesh, K., Franka, E., Abudher, A. (2011). Enteric pathogens associated with childhood diarrhea in Tripoli-Libya. The American journal of tropical medicine and hygiene, 84(6), 886-891.
Rashedul, K. (2011). Detection of Different Diarrheagenic Escherichia coli Strains by Multiplex PCR assay. Department of Microbiology (Master thesis submitted to Mymensingh Medical College, Mymensingh University), Mymensingh, Bangladesh.
Sokal, R. R. and Rohlf, F.J. (2013). Introduction to Biostatistics, 2nd ed. Dover. NewYork, 1099-1121.
Sharifi, Y., Akbari, A., & Soltan, D. (2011). Multiplex polymerase chain reaction (PCR) assay for simultaneous detection of shiga-like toxin (stx 1 and stx 2), intimin (eae) and invasive plasmid antigen H (ipaH) genes in diarrheagenic Escherichia coli. African Journal of Biotechnology, 10(9), 1522-1526.
Spano, L.C., da Cunha, K.F., Monfardini, M.V., Bergamaschi, R.D., Isabel, F., Affonso, C. (2017). High prevalence of diarrheagenic Escherichia coli carrying toxin-encoding genes isolated from children and adults in southeastern Brazil. BMC Infect Dis 17, 773 (2017).
Sperandio, V., & Pacheco, A. R. (2012). Shiga toxin in enterohemorrhagic E. coli: regulation and novel anti-virulence strategies. Frontiers in cellular and infection microbiology, 2, 81
Sudershan, R., Kumar, R. N., Kulkarni, B., Kashinath, L., Bhaskar, V., & Polasa, K. (2014). E coli pathotypes and their antibiotic resistance in young children with diarrhea in Hyderabad, India. Int Journal Curr Microbiol App Sci, 3(9), 647-654.
Toma, C., Lu, Y., Higa, N., Nakasone, N., Chinen, I., Baschkier, A., Iwanaga, M. (2003). Multiplex PCR assay for identification of human diarrheagenic Escherichia coli. Journal of clinical microbiology, 41(6), 2669-2671.
Versalovic, J., Carroll, K., Funke, G., Jorgensen, J., Landry, M., Warnock, D. (2011). Manual of clinical microbiology, ASM Press. Washington, DC.
Weinstein, M., Patel, J., Bobenchik, A. (2017). Clinical and laboratory standards institute. Performance Standards for Antimicrobial Susceptibility Testing, 27th ed.; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 296.
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2020-06-30
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Hasan, H. K., Yassin, N. A., & Eassa, S. H. (2020). Bacteriological and Molecular Characterization of Diarrheagenic Escherichia Coli Pathotypes From Children in Duhok City, Iraq. Science Journal of University of Zakho, 8(2), 52–57. https://doi.org/10.25271/sjuoz.2020.8.1.702
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