Phylogenetic Grouping and Phenotypic Detection of Extended-Spectrum β-Lactamases Among Escherichia coli From Calves and Dairy Cows in Khuzestan, Iran

Abstract

Background: Food-producing animals are under suspicion for the reservoir and colonization of ESBL (extended-spectrum beta-lactamase)-producing bacteria especially Enterobacteriaceae and therefore infection of the humans with them. The increasing reports on the ESBLs presence in the pathogenic and commensal Escherichia coli isolates have been a concern worldwide. These strains can be attributed to one of the main phylogenetic groups and subgroups. Several studies have shown the relationship between the phylogeny and antimicrobial resistance of E. coli strains. Objectives: The aim of this study was to analyze the phylogenetic group of ESBL-producing E. coli and detect its phenotype using the multiplex polymerase chain reaction (PCR) and combined disk method. Materials and Methods: Two hundred five E. coli fecal isolates were obtained from 103 calves (90 healthy and 13 diarrheic) and 102 dairy cows (healthy) from 8 farms in Khuzestan province, Iran. The triplex PCR method was used to allocate the E. coli isolates based on the presence or absence of 3 genes (chuA, yjaA, and tspE4.C2) to yield 4 definite phylogenetic groups and 7 subgroups. Phenotypic ESBL-producing E. coli was determined using the double disk diffusion method according to the manufacturer's instructions and Clinical & Laboratory Standards Institute (CLSI) guidelines. Results: A total of 65.04% and 22.3% of isolates from calves and 70.5% and 20.5% of isolates from dairy cows belonged to phylogroups B1 and A, respectively. In addition, no isolate from the diarrhoeic calves was found to belong to group B2 and subgroups D2 and A0. A low prevalence (2/205 isolates, 0.97%) of ESBL-producing E. coli was found only in the samples of dairy cows which belonged to the phylogenetic group A and phylogenetic subgroup A1. There was no statistically significant relationship between the phylogenetic group and the production of ESBLs (P = 0.11). There was also no difference between the E. coli isolates from calves and dairy cows in the production of ESBLs (P = 0.5). Conclusion: There was no statistically significant relationship between the phylogenetic group and the production of ESBLs (P = 0.11). There was also no difference between E. coli isolates from calves and dairy cows in the production of ESBLs (P = 0.5). Based on these results, there is a low prevalence of ESBL-producing E. coli in the dairy farms of Khuzestan province. However, further large scale investigations are necessary to control the antibiotic resistance in the human and animal foodstuff.

Carbapenem Resistance Profiles of Pathogenic Escherichia coli in Uganda

“…and D2. Pathogenic intestinal E. coli (DEC) mainly belong to Phylogenetic groups A, B1, and D, commensals to the groups A and B1, and strains usually belong to the groups B2 and D [13,14]. In our study, phylogenetic analysis predominantly clustered E. coli clinical isolates into B2 followed by A, B1 and D and this corroborates with ndings from previous studies [67][68][69].…”

“…Ampli cation was conducted using the following PCR conditions; initial denaturation at 94 °C for 5 minutes, then 30 cycles performed at 94 °C for 5 seconds, 54 °C for 10 seconds, 72 °C for 30 second with a nal extension step at 72 °C for 5 minutes. Phylogenic groups and subgroups were assigned depending on chuA, yjaA, and TspE4.C2 gene combinations [13,14], Table 5. CGC GCC AAC AAA GTA TTA CG and L-ETEC (2.0%).…”

Carbapenem resistance profiles of Pathogenic Escherichia coli in Uganda

“…Phylogenetic analysis revealed that E. coli isolates obtained from the four tertiary hospitals located in the Central region (MNRH), Western region (MBRRH), South Western Region (KIU-TH) and Eastern Region (MRRH) belonged to the phylogroups A B1, B2 and D and phylogenetic sub groups A0, A1, B22, B23, D1 and D2. Pathogenic intestinal E. coli (DEC) mainly belong to Phylogenetic groups A, B1, and D, commensals to the groups A and B1, and strains usually belong to the groups B2 and D [13,14]. In our study, phylogenetic analysis predominantly clustered E. coli clinical isolates into B2 followed by A, B1 and D and this corroborates with ndings from previous studies [67][68][69].…”

“…Furthermore, PCR analysis clusters E. coli strains into A, B1, B2, and D phylogenetic groups due to the presence of the chuA and yjaA genes as well as TSPE4.C2 DNA fragment [13]. The intestinal pathogenic E. coli strains belong to groups A, B1 and D, extraintestinal pathogenic E. coli strains generally follow under groups B2 and D, while commensal E. coli strains to groups A and B1 [13,14] High levels antibiotic resistance in Enterobacteriaceae is of great concern to the healthcare system [15,16]. Escherichia coli like other Enterobacteriaceae has evolved to acquire different mechanisms of antibiotic resistance which confer protection to lethal doses of different classes of antibiotics.…”

Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in cattle production – a threat around the world

“…The second continent with the highest number of countries reporting E-ESBL in cattle is Asia, which shows description of E-ESBL in 13 countries (China, Hong Kong, India, Indonesia, Iran, Israel, Japan, Lebanon, Malaysia, Saudi Arabia, South Korea, Thailand and Taiwan), highlighting China and India which are the fourth and fifth largest cattle producers in the world (Adler et al, 2015;Ali et al, 2017;Barzan et al, 2017;Diab et al, 2016;Hassan et al, 2015;Hinthong et al, 2017;Kamaruzzaman, 2015;Koovapra et al, 2016;Ohnishi et al, 2013a;Su et al, 2016;Sudarwanto et al, 2016;Tark et al, 2017). They have different roles in the import and export scenario, as India is the world's largest exporter of cattle and China the largest importer, since its internal production is not sufficient for internal consumption (USDA et al, 2017).…”