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Molecular and Phenotypic Characterization of Escherichia coli Associated with Granulomatous Colitis of Boxer Dogs.

ABSTRACT: Invasive Escherichia coli is causally associated with granulomatous colitis (GC) of Boxer dogs and French Bulldogs. The virulence determinants of GC E. coli are unclear. E. coli isolated from 16 GC (36 strains) and 17 healthy control (HC: 33 strains) dogs were diverse in phylogeny, genotype, and serotype and lacked diarrheagenic genes. Genes encoding type II (gsp), IV (traC), and VI (hcp) secretion systems, long polar fimbriae (lpfA154/141), and iron acquisition (fyuA, chuA) were frequent in GC and HC. E. coli from 14/15 GC and 10/11 HC invaded Caco-2 better than non-pathogenic E. coli strain DH5?, with invasion correlated with motility and presence of chuA and colV. E. coli from all GC and 10/11 HC survived better than DH5? in J774 macrophages, with adherent-invasive E. coli (AIEC) in 60% GC and 73% HC. AIEC replicated in monocyte derived macrophages from a GC Boxer with CD48/SLAM risk haplotype but not the HC. Fluroquinolone resistant E. coli were less motile and invasive than fluoroquinolone sensitive (p < 0.05), and only 1/8 resistant strains met criteria for AIEC. In conclusion GC E. coli are diverse, resemble extraintestinal pathogenic E. coli (ExPEC), including AIEC, and can replicate in GC-susceptible macrophages. They are likely resident pathosymbionts that can opportunistically persist within macrophages of a GC-susceptible dog.

SUBMITTER: Dogan B

PROVIDER: S-EPMC7559917 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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Molecular and Phenotypic Characterization of Escherichia coli Associated with Granulomatous Colitis of Boxer Dogs.

Dogan Belgin B Zhang Shiying S Kalla Sarah E SE Dogan Esra I EI Guo Cindy C Ang Chelston R CR Simpson Kenneth W KW

Antibiotics (Basel, Switzerland) 20200825 9

Invasive Escherichia coli is causally associated with granulomatous colitis (GC) of Boxer dogs and French Bulldogs. The virulence determinants of GC E. coli are unclear. E. coli isolated from 16 GC (36 strains) and 17 healthy control (HC: 33 strains) dogs were diverse in phylogeny, genotype, and serotype and lacked diarrheagenic genes. Genes encoding type II (gsp), IV (traC), and VI (hcp) secretion systems, long polar fimbriae (lpfA154/141), and i ...[more]

PMID: 32854367

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Project description:BackgroundEradication of intramucosal Escherichia coli correlates with remission of periodic acid-Schiff-positive E coli-associated granulomatous colitis (GC). Treatment failures attributed to multidrug resistant (MDR) bacteria necessitate alternative approaches.Hypothesis/objectivesDetermine clinical outcome of E coli-associated GC in dogs treated based on antimicrobial susceptibility profiling and characterize E coli phylogeny and resistance mechanisms.AnimalsTwenty Boxers and 4 French Bulldogs with E coli-associated GC.MethodsCulture, antimicrobial susceptibility profiling, and molecular characterization of E coli were performed and response to treatment was evaluated.ResultsInitial biopsy sample culture yielded fluoroquinolone-sensitive (FQ-S) E coli from 9/24 dogs and fluoroquinolone-resistant (FQ-R) E coli from 15/24. All but 1 FQ-R E coli were MDR with susceptibility to macrophage-penetrating antimicrobials restricted to carbapenems in 13/15 dogs. Of 22/24 treated based on susceptibility profiling, 8/9 FQ-S dogs had complete initial clinical response (CR) during fluoroquinolone (FQ) treatment, whereas 9/13 FQ-R dogs had complete or partial response (PR) during meropenem or doxycycline treatment. In 5/9 FQ-S and 12/13 FQ-R dogs with follow-up ?3 months, CR was sustained in 5/5 FQ-S (median, 25?months; range, 4-46) whereas 6/12 FQ-R had long-term CR (median, 59?months; range 15-102), 4/12 PR (median, 19?months; range, 5-65), and 2/12 had no response (NR). Four dogs with long-term follow-up died within 4?years of diagnosis, including 2 euthanized for refractory colitis. Escherichia coli were genetically diverse. Fluoroquinolone resistance was associated with mutations in gyrA and parC, with plasmid-mediated resistance less common.Conclusions and clinical importanceAntimicrobial treatment guided by susceptibility profiling was associated with positive long-term outcomes in >80% of cases. Fluoroquinolone-resistance was widespread and not clonal. Further study is required to optimize treatment for dogs with MDR E coli-associated GC.

Project description:BackgroundResistance to cephalosporins in Enterobacteriaceae is mainly due to the production of extended-spectrum beta-lactamase (ESBL). Little is known about ESBL-producing bacteria in Bangladesh. Therefore, the study presents results of phenotypic and molecular characterization of ESBL-producing Escherichia coli from hospitals in Bangladesh.MethodsA total of 339 E. coli isolated from patients with urinary tract and wound infections attending three different medical hospitals in urban and rural areas of Bangladesh between 2003-2007 were screened for ESBL-production by the double disk diffusion test. Isolates with ESBL-phenotype were further characterized by antibiotic susceptibility testing, PCR and sequencing of different β-lactamase and virulence genes, serotyping, and XbaI-macrorestriction followed by pulsed-field gel electrophoresis (PFGE).ResultsWe identified 40 E. coli with ESBL phenotype. These isolates were resistant to ceftriaxone, ceftazidime, cefotaxime, aztreonam, cefepime, and nalidixic acid but remained susceptible to imipenem. All but one isolate were additionally resistant to ciprofloxacin, and 3 isolates were resistant to cefoxitin. ESBL genes of blaCTX-M-1-group were detected in all isolates; blaTEM-type and blaOXA-1-type genes were detected in 33 (82.5%) and 19 (47.5%) isolates, respectively. Virulence genes that are present in diarrhoeagenic E. coli were not found. Class-1 integron was present in 20 (50%) isolates. All the ESBL-producing E. coli isolates harbored plasmids ranging between 1.1 and 120 MDa. PFGE-typing revealed 26 different pulsotypes, but identical pulsotype showed 6 isolates of serotype O25:H4.ConclusionThe prevalence of multidrug-resistant ESBL-producing E. coli isolates appears to be high and the majority of the isolates were positive for blaCTX-M. Although there was genetic heterogeneity among isolates, presence of a cluster of isolates belonging to serotype O25:H4 indicates dissemination of the pandemic uropathogenic E. coli clone in Bangladesh.

Project description:Many studies have examined avian pathogenic Escherichia coli (APEC) from commercial broilers but few have examined isolates from native chickens. This study compared APEC isolates from commercial broilers and native chickens in regard to the phylogenetic group and the phenotypic and genotypic antimicrobial resistance profiles. From 100 suspect colibacillosis cases in both commercial broilers and native chickens, a total of 90 broiler isolates and 42 native chicken isolates were identified as E. coli by biochemical tests. Phylogenetic grouping revealed that 90 broiler APEC isolates belonged to A group (5.56%), B1 group (22.22%), B2 group (31.11%), and D group (41.11%). The 42 native chicken APEC isolates belonged to A group (35.71%), B1 group (26.19%), B2 group (30.95%), and D group (7.14%). The difference in the allocation to groups A and D of the 2 isolate types was significant (P < 0.05). The APEC broiler isolates had a significantly higher multidrug-resistant (MDR) rate (80%) than the native chicken isolates (14.29%) (P < 0.05). The APEC broiler isolates demonstrated significantly higher resistance rates than the native chicken isolates for amoxicillin (98.89%; 78.57% respectively), chloramphenicol (42.2%; 9.5%), enrofloxacin (68.9%; 7.1%), gentamicin (11.1%; 0%), nalidixic acid (72.2%; 7.1%), sulfamethoxazole + trimethoprim (45.6%; 2.4%), and tetracycline (88.9%; 76.2%) (P < 0.05). The APEC broiler isolates had a significantly higher presence compared with the native chicken isolates of the following resistance genes:- by blaTEM (43.3%; 21.4%, respectively), cml-A (34.4%; 2.4%), tetA (76.7%; 40.5%), tetB (26.7%; 0%), sul2 (23.3%; 14.3%), and dhfrI (13.3%; 0%) (P < 0.05). The qnrB and qnrS genes were detected (12.16%; 72.97% respectively), in the APEC broiler isolates resistant to nalidixic acid and/or enrofloxacin while only qnrS genes was detected in all 3 APEC native chicken isolates. Regarding the point mutations of gyrA and parC, all isolates were positive to gyrA83S, gyrA87D, gyrA87L, gyrA87NY, parC80S and parC80I except that gyrA83S was not present in 20 APEC broiler isolates. Antimicrobial stewardship programs should be targeted at the backyard poultry sector as well as the commercial poultry sector.

Dataset Information

Molecular and Phenotypic Characterization of Escherichia coli Associated with Granulomatous Colitis of Boxer Dogs.

Publications

Molecular and Phenotypic Characterization of <i>Escherichia coli</i> Associated with Granulomatous Colitis of Boxer Dogs.

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