Aims: This study investigated the molecular diversity and extended spectrum beta-lactamase resistance of diarrheagenic E. coli isolated from patients attending selected healthcare facilities in Nasarawa State, Nigeria. Place and Duration of Study: Department of Microbiology, Nasarawa State University, P.M.B 1022, Keffi, Nasarawa State, Nigeria; between December 2017 and June, 2019. Methodology: A total of 207 confirmed E. coli isolates (using standard microbiological methods) from loose stool samples of patients with suspected cases of diarrhea (69 from Federal Medical Centre Keffi [MCK] 69 from General Hospital Akwanga [GHA] and 69 from Dalhatu Araf Specialist Hospital Lafia [DASHL]) were included in this study. Results: Phenotypic detection of ESBL production by β-lactam resistant isolates was done using double disc synergy test. Molecular detection of ESBL genes in phenotypically confirmed ESBL producers was done using Polymerase Chain Reaction. Out of 56 isolates jointly resistant to cefotaxime and/or ceftazidime and ciprofloxacin from DASHL, FMCK and GHA, 53.6% (30/56) were ESBL producers, distributed in relation to the hospitals as follows: blaCTX-M in DASHL was 6(66.7%), FMCK was 11(100.0%), and GHA was 10(100.0%); blaSHV in DASHL was 8(88.9%), FMCK was 7(63.6%), and GHA was 10(100.0%), and blaTEM in DASHL was 9(100.0%), FMCK was 10(90.9%), and GHA was 10(100.0%). Also, the occurrence of blaSHV was 100.0% in GHA but 88.9% in DASHL. The detection DEC was high in DASHL (88.9%) but low inGHA (58.8%). The occurrence of ETEC was high in GHA (60.0%) while EAEC was also high in FMCK (81.8%) and GHA (70.0%). The isolates were distributed into strain A – J based on RFLP pattern and the occurrence of strain A was high in GHA (70.0%) but low in DASHL (33.3%). Conclusion: Most of the isolates were both diarrheagenic and ESBL resistant, and the predominant ESBL and pathotypes genes were blaCTX-M, blaTEM and EAEC. Further studies on molecular detection of sub-types of ESBL and sequencing of diarrheagenic pathotypes genes should be carried out.
Aims: This study investigated the phenotypic detection of extended spectrum beta-lactamase resistance of diarrheagenic E. coli isolated from diarrheic patients attending some major health facilities in Nasarawa State, Nigeria. Place and Duration of Study: Department of Microbiology, Nasarawa State University, P.M.B 1022, Keffi, Nasarawa State, Nigeria; between December, 2017 to March, 2019. Methodology: A total of 207 confirmed E. coli isolates from loose stool samples of patients with suspected cases of diarrhea (69 from Federal Medical Centre Keffi [MCK] 69 from General Hospital Akwanga [GHA] and 69 from Dalhatu Araf Specialist Hospital Lafia [DASHL]) were included in this study. Results: E. coli was isolated and identified using standard microbiological methods. The antibiotic susceptibility testing for the isolates was carried out and interpreted in accordance with Clinical and Laboratory Standards Institute protocol. Phenotypic detection of ESBL production in isolates resistant to ciprofloxacin, cefotaxime and ceftazidime) was carried out using double disc synergy test. The occurrence of E. coli was 100% in all the hospitals. Age groups 0-5 and 6-10 years have the highest occurrence than age group 35 – >45 years. Isolates from DASHL were more resistant to amoxicillin/clavulanic acid (86.9%), Streptomycin (75.0%) and sulphamethoxazole/trimethoprim (68.1%), isolates from FMCK were more resistant to amoxicillin/clavulanic acid (84.1%), sulphamethoxazole/trimethoprim (69.6%), isolates from GHA were more resistant to amoxicillin/clavulanic acid (85.5%) and sulphamethoxazole/trimethoprim (73.0%). Multiple antibiotic resistance (MAR) was observed with the order of occurrence: FMCK (98.6%) > DASHL (92.8%) > GHA (89.9%). The most common MAR index of 0.2 in DASHL was 0.4 (20.3%); FMCK was 0.4 (15.9%) and GHA was 0.3 (17.4%). The order of occurrence of classes of antibiotic resistance in E. coli isolates in DASHL was MDR (84.0%) ˃ XDR(7.2%) > PDR and NMDR (4.3%); in FMCK was MDR (91.3%) ˃ XDR(4.3%) ˃ NMDR (2.9%) and PDR(1.4%); and in GHA was MDR (88.8%) ˃ NMDR(5.8%) > XDR and PDR(2.9%). Detection rate of ESBL was 53.6% (30/207), distributed in relation to the location as DASHL (60.0%), FMCK (50.0%) and GHA (52.6%). Conclusion: Most of the isolates from the study locations were antibiotic resistance. Further studies on molecular detection of ESBL, diversity and characterization of the E. coli into pathotypes are ongoing.
Objectives: The present study reports extended-spectrum beta-lactamase (ESBL) production in E. coli isolates from poultry droppings from selected poultry farms in Keffi, Nigeria. Methods: Seventy-five (75) samples of poultry droppings were collected, and E. coli was isolated using standard microbiological methods. Antibiotic susceptibility testing and minimum inhibitory concentrations were evaluated as described by the Clinical and Laboratory Standards Institute (CLSI). Phenotypic confirmation of ESBL production by the isolates was carried out using double disc synergy test. Molecular detection of ESBL genes was carried out using Polymerase Chain Reaction (PCR) method. Results: All (100%) samples had E. coli. Antimicrobial resistance in the isolates were as follows: imipenem (12.0%), gentamicin (20.0%), cefoxitin (37.3%), cefotaxime (41.3%), ceftazidime (44.0%), ciprofloxacin (48.0%), amoxicillin/clavulanic acid (58.7%), streptomycin (92.0%), sulphamethoxazole/trimethoprim (92.0%) and ampicillin (98.7%). Joint resistance to ampicillin, sulphamethoxazole/trimethoprim-streptomycin was the commonest resistance phenotype at 10.6%. Multiple antibiotic resistance (MAR) was observed in 97.3% (73/75) of the isolates; and the most common MAR indices were 0.7 (21.9%), 0.5 (17.8%), 0.4 (16.4%), 0.8 (11.1%) and 0.3 (10.9%). Twenty three (46.9%) of the 49 cefotaxime/ceftazidime isolates were confirmed ESBL producers. Twenty-two of the 23 ESBL positive isolates (95.7%) carried the bla genes as follows: 95.5% (21/22) for blaSHV; 68.2% (15/22) for blaTEM; and 50.0% (11/22) for blaCTX-M. Eleven (50%) of the 22 isolates carried two bla genes (blaSHV and blaCTX-M, blaTEM and blaCTX-M and blaTEM and blaSHV). Conclusion: The E. coli isolates were less resistant to imipenem, gentamicin and cefoxitin; most isolates were MAR, with resistance to 7 antibiotics being the most predominant. In addition, the blaSHV gene was the most common ESBL gene detected in confirmed ESBL-producing E. coli isolates.
Aims: This study investigates and reports the production of extended spectrum beta-lactamase in Escherichia coli isolates in poultry droppings sourced from selected poultry farms in Karu, Nigeria Study Design: Cross sectional study Place and Duration of Study: Department of Microbiology, Nasarawa State University, Keffi, between August 2019 and February 2020. Methodology: Escherichia coli was isolated from the samples using standard cultural and microbiological methods. Antibiotic susceptibility testing and minimum inhibitory concentrations were evaluated as described by the Clinical and Laboratory Standards Institute (CLSI). The detection of ESBL production in E. coli isolates was carried out using double disc synergy test. In addition, molecular detection of ESBL genes was carried out using Polymerase Chain Reaction (PCR) method. Results: All (100%) samples collected had E. coli. Antibiotic resistances in the isolates in decreasing order were as follows: ampicillin (96.7%), streptomycin (94.4%), sulphamethoxazole /trimethoprim (87.8%), amoxicillin/ clavulanic acid (61.1%), gentamicin (52.2%), ciprofloxacin (40.0%), ceftazidime (35.6%), cefotaxime (31.1%), imipenems (22.2%), cefoxitin (13.3%). The commonest antibiotic resistant phenotype was AMP-SXT-S-CTX-CN (8.8%). Multiple antibiotic resistance (MAR) was observed in 92.2% (83/90) of the isolates with the common MAR indices being 0.5 (26.5%), 0.6 (19.2%), 0.4 (13.2%) and 0.9 (10.8%). Fifty nine of the eighty beta-lactam resistant isolates (73.7%) were confirmed ESBL producers. 55 of the 59 ESBL positive isolates (93.2%) carried bla genes as follows: blaSHV (50/55, 90.9%), blaTEM (31/55, 56.3%) and blaCTX-M (46/55, 83.6%). Thirty six (65.5%) of the 55 isolates carried two bla genes (blaSHV and blaTEM, blaTEM and blaCTX-M, and blaCTX-M and blaSHV). Conclusion: The E. coli isolates showed lower resistances to cefoxitin, imipenem, cefotaxime, ceftazidime, and ciprofloxacin and most isolates were MAR, with resistance to 5 antibiotics being the most predominant. In addition, blaSHV gene was the most common ESBL gene detected in the confirmed ESBL-producing E. coli isolates.
Aims: This study investigated the antimicrobial resistance profile and extended spectrum beta-lactamase resistance genes of Escherichia coli isolated from droppings of from selected poultry farms in Nasarawa, Nigeria. Study Design: Investigative Place and Duration of Study: Department of Microbiology, Nasarawa State University, Keffi, between November 2019 and February 2020. Methodology: A total of 90 samples from poultry droppings were collected from selected farms. Escherichia coli was isolated from the samples using standard cultural and microbiological methods. Antibiotic susceptibility testing and minimum inhibitory concentrations were evaluated as described by the Clinical and Laboratory Standards Institute (CLSI). The detection of extended-spectrum beta-lactamase (ESBL) production in E. coli isolates was carried out using double disc synergy test. In addition, molecular detection of ESBL genes was carried out using Polymerase Chain Reaction (PCR) method. Results: The prevalence of E. coli was 100%. Antibiotic resistances of E. coli were recorded as follows: streptomycin (S: 94.4%), sulphamethoxazole / trimethoprim (SXT: 90.0%), ampicillin (AMP: 88.9%), gentamicin (CN: 68.9%), amoxicillin/clavulanic acid (AMC: 55.6%), ciprofloxacin (CIP: 41.1%), cefoxitin (FOX: 35.6%), ceftazidime (CAZ: 34.4%), cefotaxime (CTX: 22.2%), and imipenems (IPM: 17.8%). The most common antibiotic resistant resistance phenotype was AMP-CTX-CAZ-CIP-CN (11.1%). Multiple antibiotic resistance (MAR) was observed in 97.7% (88/90) of the isolates, with the common MAR index being 0.5 (33.3%). Twenty five of the thirty beta-lactam resistant isolates (83.3%) were confirmed ESBL producers. The 25 ESBL positive isolates carried bla genes as follows: blaTEM (11/25, 44.0%) and blaCTX-M (18/25, 72.0%). blaSHV was not found in any isolate. Conclusion: E. coli isolated from the droppings of selected poultry farms in Nasarawa were less resistant to imipenem, cefotaxime, ceftazidime and cefoxitin in the study location. This implies that the antibiotics are useful in the treatment of infection caused by E. coli. Also, ESBL-positive E. coli isolates harbored ESBL genes, with blaCTX-M as the most common.
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