Isolation of Shiga toxin-producing strains of Escherichia coli from beef and swine carcasses and the characterization of their genes


Keywords: STEC; beef carcasses; swine carcasses; stx1; stx2; eae genes; multiples PCR

Abstract

Escherichia coli is part of the normal microflora of the intestinal tract of humans and warm-blooded animals, but its presence in raw material and food of animal origin is considered as fecal contamination and can be very dangerous for consumers. The determination of the number of E. coli in raw material and food is important because among them can be pathogenic strains. The most dangerous strains are considered enterohemorrhagic E. coli as a causative agent of severe bloody diarrhea and hemorrhagic uremic syndrome in humans through the production of Shiga-toxin, which is the main virulence factor, responsible for disease. The aim of this study was to identify the prevalence of Shiga toxin-producing strains of E. coli (STEC) from swabs of beef and swine carcass in slaughterhouses in Ukraine and characterize their genes, which are responsible for pathogenic properties. A total of 230 samples of swabs from beef (130) and swine (100) carcasses were obtained from 5 slaughterhouses in Ukraine between 2012 and 2015. Samples of swabs from carcasses were randomly selected at the final point of the process after the final washing of the carcass from the following areas: distal hind limb, abdomen (lateral and medial) from swine carcasses, brisket, flank and flank groin areas from beef carcasses. All samples were examined by culture-dependent method, after that each positive isolate of STEC was analyzed by multiplex PCR to detect the stx1, stx2, and eae genes. Out of 230 collected samples, seven (7.2%) were contaminated with STEC. The highest prevalence of STEC was found in swabs from beef carcasses (8.1%) in comparison to swabs from swine carcasses (5.7%). The stx1 gene was the predominant gene detected in all STEC positive samples. The eae gene was found in one of the examined isolates from beef carcass. Three isolates from swabs of beef carcass carried both stx1 and stx2 genes, one isolate showed association between stx1 and eae genes, one isolate was positive for stx1 gene only. In swabs from swine carcasses (2 isolates) stx1 and stx2 genes were presented simultaneously. The results of this study suggested that fresh raw meat could be a potential vehicle for transmission of the Shiga toxin-producing strain of E. coli to humans. This is the first report of STEC prevalence in beef and swine carcasses in Ukraine and these data will be valuable for microbiological risk assessment and help the appropriate services to develop strategies to mitigate health risk.

References

Abdissa, R., Haile, W., Fite, A. T., Beyi, A. F., Agga, G. E., Edao, B. M., Tadesse, F., Korsa, M. G., Beyene, T., Beyene, T. J., De Zutter, L., Cox, E., & Goddeeris, B. M. (2017). Prevalence of Escherichia coli O157:H7 in beef cattle at slaughter and beef carcasses at retail shops in Ethiopia. MC Infectious Diseases, 17(1), 277–283.


Anu, P. J., Latha, C., Vinodkumar, K., Vinod, V. K., & Sathu, T. (2018). An important approach for control of Enterohaemorrhagic Escherichia coli by identification of contaminating sources in beef production line. International Journal of Current Microbiology and Applied Sciences, 7(1), 1921–1929.


Awadallah, M. A., Ahmed, H. A., Merwad, A. M., & Selim, M. A. (2016). Occurrence, genotyping, shiga toxin genes and associated risk factors of E. coli isolated from dairy farms, handlers and milk consumers. The Veterinary Journal, 217, 83–88.


Bardasi, L., Taddei, R., Fiocchi, I., Pelliconi, M. F., Ramini, M., Toschi, E., & Merialdi, G. (2017). Shiga toxin-producing Escherichia coli in slaughtered pigs and pork products. Italian Journal of Food Safety, 6, 6579–6584.


Beyi, A. F., Fite, A. T., Tora, E., Tafese, A., Genu, T., Kaba, T., Beyene, T. J., Beyene, T., Korsa, M. G., Tadesse, F., Zutter, L., Goddeeris, B. M., & Cox, E. (2017). Prevalence and antimicrobial susceptibility of Escherichia coli O157 in beef at butcher shops and restaurants in central Ethiopia, BMC Microbiology, 17, 49–55.


Bonardi, S., Alpigiani, I., Tozzoli, R., Vismarra, A., Zecca, V., Greppi, C., Bacci, C., Bruini, I., & Brindani, F. (2015). Shiga toxin-producing Escherichia coli O157, O26 and O111 in cattle faeces and hides in Italy, Veterinary Record Open, 2, 1–10.


Hara-Kudo, Y., Konishi, N., Ohtsuka, K., Iwabuchi, K., Kikuchi, R., Isobe, J., Yamazaki, T., Suzuki, F., Nagai, Y., Yamada, H. and Tanouchi, A. (2016). An interlaboratory study on efficient detection of Shiga toxin-producing Escherichia coli O26, O103, O111, O121, O145, and O157 in food using real-time PCR assay and chromogenic agar. International Journal of Food Microbiology, 230, 81–88.


Haugum, K., Brandal, L. T., Lindstedt, B. A., Wester, A. L., Bergh, K., & Afseta, J. E. (2014). PCR-based detection and molecular characterization of Shiga toxin producing Escherichia coli strains in a routine microbiology laboratory over 16 years. Journal of Clinical Microbiology, 52(9), 3156–3163.


Jeon, S. J., Elzo, M., Dilorenzo, N., Lamb, G. C., & Jeong, K. C. (2013). Evaluation of animal genetic and physiological factors that affect the prevalence of Escherichia coli O157 in cattle. PLoS One, 8(2), 1–9.


Ju, W., Shen, J., Li, Y., Toro, M. A., Zhao, S., Ayers, S., Najjar, M. B., & Meng, J. (2012). Non-O157 Shiga toxin-producing Escherichia coli in retail ground beef and pork in the Washington D.C. area. Food Microbiology, 32, 371–377.


Kavaliauskiene, S., Dyve Lingelem, A. B., Skotland, T., & Sandvig, K. (2017). Protection against Shiga toxins. Toxins (Basel), 9(2), e44.


Lozinak, K. A., Jani, N., Gangiredla, J., Patel, I., Elkins, C. A., Hu, Z., Kassim, P. A., Myers, R. A., & Laksanalamai, P. (2016). Investigation of potential shiga toxin producing Escherichia coli (STEC) associated with a local foodborne outbreak using multidisciplinary approaches. Food Science, 5(3), 163–168.


Majowicz, S. E., Scallan, E., Jones-Bitton, A., Sargeant, J. M., Stapleton J., Angulo, F. J., Yeung, D. H., & Kirk, M. D. (2014). Global incidence of human Shiga toxin-producing Escherichia coli infections and deaths: A systematic review and knowledge synthesis. Foodborne Pathogens and Disease, 11, 447–455.


Merwad, A., Gharieb, R., & Saber, T. (2014). Occurrence of shiga toxin-producing Escherichia coli in lactating cows and in contact workers in Egypt: Serotypes, virulencegenes and zoonotic significance. Life Science Journal, 11(5), 563–571.


Microbiology of the food chain. Carcass sampling for microbiological analysis (2015). European Centre for Disease Prevention and Control. Escherichia coli factsheet Stockholm: ECDC. ISO 17604:2015.


Momtaz, H., & Jamshidi, A.(2013). Shiga toxin-producing Escherichia coli isolated from chicken meat in Iran: Serogroups, virulence factors, and antimicrobial resistance properties. Poultry Science, 92, 1305–1313.


Omari, K. E., Kassaa, I. A., Kara-Ali, H., Dabboussi, F., & Hamze, M. (2018). Prevalence of E. coli O157:H7 in raw minced beef at slaughterhouses in Tripoli, Lebanon. Virology and Immunology Journal, 2(4), 2–4.


Omoruyi, I. M., Uwadiae, E., Mulade, G., & Omoruku, E. (2018). Shiga toxin producing strains of Escherichia coli (STEC) associated with beef products and its potential pathogenic effect. Microbiology Research Journal International, 23(1), 1–7.


Premarathne, J. M. K. J. K., New, C. Y., Ubong, A., Nakaguchi, Y., Nishibuchi, M., & Son, R. (2017). Risk of Escherichia coli O157:H7 infection linked to the consumption of beef. Food Research, 1(3), 67–76.


Puttalingamma, V., Shylaja, R., Batra, H. V., & Bawa, A. S. (2012). A novel multiplex PCR system for the detection of virulence associated genes of E. coli O157:H7 from food system. Recent Research in Science and Technology, 4(5), 36–40.


Rani, S., Singh, Y., Gulati, B. R., & Khurana, S. K. (2017). Occurrence of enterohaemorrhagic Escherichia coli in buffalo meat. Journal of Experimental Biology and Agricultural Sciences, 5(2), 208–214.


Rantsiou, K., Alessandria, V., & Cocolin, L. (2012). Prevalence of Shiga toxin-producing Escherichia coli in food products of animal origin as determined by molecular methods. International Journal of Food Microbiology, 154, 37–43.


Rashid, M., Kotwal, S. K., Malik, M. A., & Singh, M. (2013). Prevalence, genetic profile of virulence determinants and multidrug resistance of Escherichia coli isolates from foods of animal origin. Veterinary World, 6, 139–142.


Ray, B., & Bhunia, A. (2014). Fundamental food microbiology. CRC Rress, Taylor & Francis Group, New York.


Shuhong, Z., Xuemei, Z., Qingping, W., Jumei, Z., Xiaoke, X., & Haigang, L., (2015). Prevalence and characterization of Escherichia coli O157 and O157:H7 in retail fresh raw meat in South China. Annals of Microbiology, 65, 1993–1999.


Smith, J. L., Fratamico, P. M., & Gunther, N. W. T. (2014). Shiga toxin-producing Escherichia coli. Advances in Applied Microbiology, 86, 145–197.


Soledad-Cadona, J., Bustamante, A. V., González, J., & Mariel-Sanso, A. (2018). Pathogenicity islands distribution in non-O157 Shiga toxin-producing Escherichia coli (STEC). Genes, 9, 81–99.


Stromberg, Z. R., Redweik, G. A. J., & Mellata, M. (2018). Detection, prevalence, and pathogenicity of Non-O157 shiga toxin-producing Escherichia coli from cattle hides and carcasses. Foodborne Pathogens and Disease, 15(3), 119–131.


Sudershan, R. V., Naveenkumar, R., Kashinath, L., Bhaskar, V., Polasa, K. (2014). Foodborne infections and intoxications in Hyderabad India. Epidemiolody Research International, 10, 1–5.


Tafida, S. Y., Kwaga, J. K. P., Bello, M., Kabir, J., Umoh, V. J., Yakubu, S. E., & Nok, A. J. (2014). Occurrence of Escherichia coli 0157 in retailed-beef and related meat products in Zaria, Nigeria. Food and Nutrition Sciences, 5, 481–487.


Taye, M., Berhanu, T., Berhanu, Y., Tamiru, F., & Terefe, D. (2013). Study on carcass contaminating Escherichia coli in apparently healthy slaughtered cattle in Haramaya University Slaughter House with special emphasis on Escherichia coli O157:H7. Journal of Veterinary Science and Technology, 4, 132.


Troz-Williams, L. A., Mercer, N. J., Walters, L. M., Maki, A. M., & Johnson, R. P. (2012). Pork implicated in a Shiga toxin-producing Escherichia coli O157:H7 outbreak in Ontario, Canada. Canadian Journal of Public Health, 103, 322–326.


Tseng, M., Fratamico, P. M., Bagi, L., Manzinger, D., & Funk, J. A. (2014). Shiga toxin-producing E. coli (STEC) in swine: Prevalence over the finishing period and characteristics of the STEC isolates. Epidemiology and Infection, 8, 1–10.


Vijayan, C., Ajaykumar, V. J., Bhattacharya, A., & Bhanurekka, V. (2017). Detection of enterohaemorrhagic E. coli O157: H7 from beef and chevon sold in and around Puducherry. Journal of Entomology and Zoology Studies, 5(6), 1395–1403.

Published
2018-04-23
How to Cite
BerhilevychO. М., Kasianchuk, V. V., Deriabin, O. M., & Kukhtyn, M. D. (2018). Isolation of Shiga toxin-producing strains of Escherichia coli from beef and swine carcasses and the characterization of their genes. Regulatory Mechanisms in Biosystems, 9(2), 275-780. https://doi.org/https://doi.org/10.15421/021840

Most read articles by the same author(s)

Obs.: This plugin requires at least one statistics/report plugin to be enabled. If your statistics plugins provide more than one metric then please also select a main metric on the admin's site settings page and/or on the journal manager's settings pages.