Abstract
The avian pathogenic Escherichia coli (APEC) strains are the chief etiology of colibacillosis worldwide. The present study investigated the circulating phylotypes, existence of virulence genes (VGs), and antimicrobial resistance (AMR) in 392 APEC isolates, obtained from 130 samples belonged to six farms using both phenotypic and PCR-based molecular approaches. Congo red binding (CRB) assay confirmed 174 APEC isolates which were segregated into ten, nine, and eight distinct genotypes by RAPD assay (discriminatory index, DI = 0.8707), BOX-PCR (DI = 0.8591) and ERIC-PCR (DI = 0.8371), respectively. The combination of three phylogenetic markers (chuA, yjaA and DNA fragment TspE4.C2) classified APEC isolates into B23 (37.36%), A1 (33.91%), D2 (11.49%), B22 (9.20%), and B1 (8.05%) phylotypes. Majority of the APEC isolates (75–100%) harbored VGs (ial, fimH, crl, papC, and cjrC). These VGs (papC and cjrC) and phylotypes (D2 and B2) of APEC had significant (p = 0.004) association with colibacillosis. Phylogenetic analysis showed two distinct clades (clade A and clade B) of APEC, where clade A had 98–100% similarity with E. coli APEC O78 and E. coli EHEC strains, and clade B had closest relationship with E. coli O169:H41 strain. Interestingly, phylogroups B2 and D2 were found in the APEC strains of both clades, while the strains from phylogroups A1 and B1 were found in clade A only. In this study, 81.71% of the isolates were biofilm formers, and possessed plasmids of varying ranges (1.0 to 54 kb). In vitro antibiogram profiling revealed that 100% isolates were resistant to ≥3 antibiotics, of which 61.96%, 55.24%, 53.85%, 51.16% and 45.58% isolates in phylotypes B1, D2, B22, B23, and A1, respectively, were resistant to these antimicrobials. The resistance patterns varied among different phylotypes, notably in phylotype B22, showing the highest resistance to ampicillin (90.91%), nalidixic acid (90.11%), tetracycline (83.72%), and nitrofurantoin (65.12%). Correspondence analysis also showed significant correlation among phylotypes with CRB (p = 0.008), biofilm formation (p = 0.02), drug resistance (p = 0.03), and VGs (p = 0.06). This report demonstrated that B2 and A1 phylotypes are dominantly circulating APEC phylotypes in Bangladesh; however, B2 and D2 are strongly associated with the pathogenicity. A high prevalence of antibiotic-resistant APEC strains from different phylotypes suggest the use of organic antimicrobial compounds, and/or metals, and the rotational use of antibiotics in poultry farms in Bangladesh.
Highlights
Escherichia coli is a ubiquitous organism having the fabulous adaptive ability in diverse ecological niches including the intestine of animals and humans [1]
According to the microbiological analysis, 392 isolates were obtained through selective identification in Eosin Methylene Blue (EMB) and MacConkey agar, and biochemical tests followed by Congo red binding (CRB) assay
Our results demonstrated that avian pathogenic E. coli (APEC) phylotypes distribution differed significantly (p = 0.002) across the study areas
Summary
Escherichia coli is a ubiquitous organism having the fabulous adaptive ability in diverse ecological niches including the intestine of animals and humans [1] This pathogen can induce enteric and extraintestinal infections [1,2]. Several studies revealed the incidence of various phylotypes of APEC strains in combinations of virulence-associated genes (VGs) [7,26,27] These virulence factors are associated with various virulence genetic markers, such as P fimbriae structural subunit (papA) and P fimbriae assembly (papC) [28], fimA (encoding type 1 fimbriae), bundle-forming pilus (bfp), aerobactin iron uptake system (aer) [29], crl (curli fimbriae), and many more which are linked to zoonotic concern [19]. The role of curli fimbriae which encodes for crl and csgA genes in the pathogenesis is poorly elucidated, though these genes facilitate the adherence of APEC strains to fibronectin and laminin [30,31,32]
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