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Prevalence of Shiga Toxin-Producing O157 and Non-O157 Escherichia coli in Anatolian Buffaloes (Bubalus bubalis)
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Abstract
Shiga toxin producing Escherichia coli (STEC) serotypes are recognized as potentially important food-borne pathogens for humans. Ingestion of E. coli contaminated food is largely known to originate from livestock. Cattle and sheep herds hold the majority of agricultural revenue in Türkiye but Anatolian water buffaloes have often been underestimated for foodborne pathogens. The aim of this study is to determine virulence genes harboring E. coli O157 and six major non-O157 STEC (O26, O45, O103, O111, O121, and O145) serotypes in feces of healthy Anatolian water buffaloes by using multiplex PCR (mPCR) method. Of the collected 458 fecal samples from healthy live animals, we have performed virulence and serotype targeting mPCR following direct DNA extraction from collected samples. Results indicate that there is 0.9% of O157 prevalence while six major non-O157 E. coli have not been identified. The characterization results of the virulence genes also showed that eae is most prevalent (5.7%) followed by ehxA (3.9%) and stx1 (3.1%). In this study, we have shown Anatolian buffaloes might have a relationship with other O-type E. coli strains. Non-O157 STECs, which are often disregarded in both animals and humans, should be investigated. As a consequence, gaining regional or national data collection will allow to implement better effective diagnosis and treatment options.
Keywords: E. coli O157; non-O157; STEC; Anatolian Buffaloes.
Introduction
Escherichia coli (E. coli) consists of a diverse and large group of bacterial organisms residing within the gut of humans and animals (1). Although most E. coli variants are harmless or result in asymptomatic colonization, some pathogenic variants can cause gastroenteritis, including hemorrhagic colitis (HC) or extra-intestinal infections such as urinary tract infections, hemolytic uremic syndrome (HUS) or other severe systematic infections including pneumonia (2).
Diarrhea-causing pathogenic E. coli variants are identified under several groups according to their O-type serogroup classification and pathogenesis with clinical implications: entero-aggregative E. coli, entero-pathogenic E. coli (EPEC), diffuse-adherent E. coli, entero-invasive E. coli, and entero- haemorrhagic E. coli (3). Shiga toxin-producing E. coli (STEC) agents are mostly associated with the entero-haemorrhagic group of variants but several types of diarrheagenic E. coli pathovariants other than O157 can also produce shiga toxin (4). In a public outbreak that emerged in Germany in 2011, it was found that non-O157 shiga toxin-producing E. coli O104 was responsible for mass public infections. Among the infected patients (3842 cases) in Germany, tragedy 22.1% of people developed HUS, and the outbreak resulted in 1.4% mortality. A detailed investigation confirmed that the consumption of contaminated vegetable sprouts led to the infection (5). Many large sporadic outbreaks have emerged in the past between 1984 and 2009 but non-O157 STEC cases rarely have been reported due to the absence of identification and characterization of non-O157 STEC methods (6). In a different outbreak led by foodborne infection, it has been reported that 23 children from South Australia (1995) and 10 children from Norway (2006) developed HUS due to non-O157 STEC; O111 and O103 respectively (7, 8). As a result, we are now much more aware of the importance of both O157 and non-O157 STEC for public health issues due to unexpected sources of infection and outbreak.
E. coli strains may acquire virulence factors such as shiga toxin 1 and 2 (stx1, stx2) intimin (eae), and enterohaemolysin (ehxA). It was suspected that there was a close relationship between virulence factors and clinical manifestations. Shiga toxins are cytotoxins that inhibit protein synthesis and cause host cell death. Intimin is shared by STEC and EPEC implements which is tightly attach to epithelial cells. Therefore, causes attaching and effacing (A/E) lesions in the intestinal mucosa. Especially HC and HUS cases with severe diarrhea were determined to be closely related to STEC types carrying the eae gene (9). Enterohaemolysin is thought to provide an iron source to stimulate E. coli growth by lysis of erythrocytes and release of heme from hemoglobin (10). Hence, virulence genes incorporating E. coli strains or their combination with various virulence genes maybe closely associated with clinical symptoms of intestinal or extra-intestinal infections.
Foodborne STEC infections are caused mainly by contaminated water sources via close contact with fecal materials of farm animals, primarily cattle followed by goats, sheep, and pigs (11). Although most of the daily dietary needs of humans are primarily supplied from cattle, buffaloes are also considered as a source of meat and dairy products.
Türkiye continues breeding Anatolian buffaloes, descended from Mediterranean water buffalo, for its economic value and special aromas of dairy products. Previously, epidemiological investigation of O157 and non-O157 serogroup STEC isolation studies were carried out in farm animals including cattle and sheep in Türkiye, however there is less known about Anatolian water buffaloes as a potential source of STEC. Moreover, prevalence studies of STEC in Anatolian water buffaloes are limited. A few studies have been conducted in Anatolian water buffaloes showing the prevalence of O157:H7 STEC, but the presence of non-O157 STEC has not yet been elucidated (12-16). Besides, the importance of virulence genes harboring E. coli serotypes in relation to public health should also need to be highlighted.
The purpose of this study was to determine E. coli O157 and six major non-O157 (O26, O45, O103, O111, O121, O145) E. coli serotypes in feces of healthy Anatolian water buffaloes by characterization of encoded virulence genes: shiga toxin 1 (stx1), shiga toxin 2 (stx2), intimin (eae) and enterohaemolysin (ehxA) using a mPCR assay. [...]
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