scholarly journals Molecular Diversity and Extended Spectrum Beta-lactamase Resistance of Diarrheagenic Escherichia coli from Patients Attending Selected Health Care Facilities in Nasarawa State, Nigeria

Author(s):  
R. H. Abimiku ◽  
Y. B. Ngwai ◽  
I. H. Nkene ◽  
B. E. Bassey ◽  
P. A. Tsaku ◽  
...  

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.

Author(s):  
S. C. Tama ◽  
Y. B. Ngwai ◽  
I. H. Nkene ◽  
R. H. Abimiku

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.


Author(s):  
S. C. Tama ◽  
Y. B. Ngwai ◽  
G. R. I. Pennap ◽  
I. H. Nkene ◽  
R. H. Abimiku

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.


Author(s):  
R. H. Abimiku ◽  
Y. B, Ngwai ◽  
I. H. Nkene ◽  
B. E. Bassey ◽  
P. A. Tsaku ◽  
...  

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.


Antibiotics ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 406
Author(s):  
Zuhura I. Kimera ◽  
Fauster X. Mgaya ◽  
Gerald Misinzo ◽  
Stephen E. Mshana ◽  
Nyambura Moremi ◽  
...  

We determined the phenotypic profile of multidrug-resistant (MDR) Escherichia coli isolated from 698 samples (390 and 308 from poultry and domestic pigs, respectively). In total, 562 Enterobacteria were isolated. About 80.5% of the isolates were E. coli. Occurrence of E. coli was significantly higher among domestic pigs (73.1%) than in poultry (60.5%) (p = 0.000). In both poultry and domestic pigs, E. coli isolates were highly resistant to tetracycline (63.5%), nalidixic acid (53.7%), ampicillin (52.3%), and trimethoprim/sulfamethoxazole (50.9%). About 51.6%, 65.3%, and 53.7% of E. coli were MDR, extended-spectrum beta lactamase-producing enterobacteriaceae (ESBL-PE), and quinolone-resistant, respectively. A total of 68% of the extended-spectrum beta lactamase (ESBL) producers were also resistant to quinolones. For all tested antibiotics, resistance was significantly higher in ESBL-producing and quinolone-resistant isolates than the non-ESBL producers and non-quinolone-resistant E. coli. Eight isolates were resistant to eight classes of antimicrobials. We compared phenotypic with genotypic results of 20 MDR E. coli isolates, ESBL producers, and quinolone-resistant strains and found 80% harbored blaCTX-M, 15% aac(6)-lb-cr, 10% qnrB, and 5% qepA. None harbored TEM, SHV, qnrA, qnrS, qnrC, or qnrD. The observed pattern and level of resistance render this portfolio of antibiotics ineffective for their intended use.


2017 ◽  
Vol 5 (1) ◽  
pp. 32-38 ◽  
Author(s):  
Ujjwal Rimal ◽  
Shovana Thapa ◽  
Roshani Maharjan

Extended-spectrum β-lactamase producing E. coli and K. pneumoniae is a serious threat to the patients. These organisms are major extended spectrum beta lactamase (ESBL) producers. The objective of this study was to determine the prevalence of Extended spectrum β- lactamase producing strains of Escherichia coli and Klebsiella spp isolates from the urine sample of children visiting International Friendship Children Hospital. During the seven months, between June 2016 to December 2016, 1018 mid-stream urine samples(MSU) were collected from patients suspected of having UTI. The samples were investigated by conventional semi-quantitative culture technique and identification of E. coli and Klebsiella spp. was done by microscopy and biochemical test. Antibiotic susceptibility test of isolates was performed by modified Kirby Bauer Disc diffusion test. ESBL screening test was done by using 3rd generation Cephalosporin and confirmation done by combination disc diffusion method. Out of total 1018 MSU samples investigated, 200(19.64%) isolates of E. coli and 28(2.7%) isolates of Klebsiella spp. making a total of 228(22.39%) were found to cause significant bacteriuria. 76(33.33%) isolates, from those causing significant bacteriuria, were Multi-drug resistant organisms. Out of 228 isolates, 54(23.68%) were ESBL producers, that includes 51(25.5%) Escherichia coli and 3(12.5%) Klebsiella pneumoniae. ESBL producers were more common in in-patient (36.17%) than out-patient (20.44%). Most of the ESBL producers were resistance to amoxicillin, followed by Cotrimoxazole and Ciprofloxacin respectively. They were highly sensitive to Imipenem, Tigecycline, Amikacin, Piperacillin-Tazobactam, and Nitrofurantoin. High prevalence of ESBL producing E. coli and Klebsiella pneumoniae was found among children. Regular and routine monitoring of ESBL producing isolates is essential.Nepal Journal of Biotechnology. Dec. 2017 Vol. 5, No. 1: 32-38


2009 ◽  
Vol 20 (suppl a) ◽  
pp. 43A-48A ◽  
Author(s):  
Patricia J Baudry ◽  
Melissa McCracken ◽  
Philippe Lagacé-Wiens ◽  
Michael R Mulvey ◽  
George G Zhanel ◽  
...  

OBJECTIVE: The purpose of the present study was to determine the prevalence and molecular epidemiology of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae identified from Canadian hospitals in 2007. METHODS: Clinically significant isolates were collected as part of the Canadian Ward Surveillance Study (CANWARD 2007) from January to December 2007, inclusive, from 12 sentinel hospital centres across Canada. Minimum inhibitory concentrations were determined by broth microdilution, and putative ESBL isolates were confirmed by the Clinical and Laboratory Standards Institute disk diffusion method. Polymerase chain reaction and DNA sequencing were used to detectblaSHV,blaTEM,blaCTX-MandblaOXA-likegenes. Strains were typed using pulsed-field gel electrophoresis. RESULTS: A total of 3.4% and 1.6% ofEscherichia coliandKlebsiella pneumoniae, respectively, were identified as ESBL producers. Resistance to fluoroquinolones, doxycycline, trimethoprim/sulfamethoxazole and gentamicin occurred in 92.5% and 71.4%, 75.5% and 71.4%, 67.9% and 57.1%, and 58.5% and 57.1% of ESBL-producingE coliandK pneumoniae, respectively. A total of 90.6% and 71.4% of ESBL-producingE coliandK pneumoniaewere identified as multidrug resistant. The CTX-M type was the predominant ESBL, with CTX-M-15 as the predominant genotype. A total of 81.7% ESBL-producers carried several beta-lactamase genes. Pulsed-field gel electrophoresis revealed that the majority of ESBL producers were not genetically related (less than 80% homology). Similar patient demographics were observed among both ESBL-producingE coliand K pneumoniae. CONCLUSION: CTX-M has become the most common enzyme among both ESBL-producingE coliandK pneumoniae. The spread of ESBLproducing bacteria across Canada is polyclonal and is not due to the clonal spread of a single strain.


Antibiotics ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 747
Author(s):  
Rumyana Markovska ◽  
Petya Stankova ◽  
Temenuga Stoeva ◽  
Dobrinka Ivanova ◽  
Daniela Pencheva ◽  
...  

The gastrointestinal tract is an important reservoir of extended spectrum beta-lactamase (ESBL)/carbapenemase-producing Enterobacterales isolates. This study included patients from two Bulgarian hospitals. Overall, 98 ESBL producers (including 68 Escherichia coli and 20 Klebsiella pneumoniae isolates) were detected among 99 hospitalized patients, 212 patients at admission, and 92 hospital staff in 42.4%, 24.5%, and 4%, respectively. We observed blaCTX-M-15 in 47% of isolates, blaCTX-M-3 in 39% and blaCTX-M-14 in 11%. Three blaCTX-M-15 positive isolates were also blaKPC-2 positive. High transferability was detected for blaCTX-M-3 carrying plasmids (55%) with L/M and I1 replicon plasmids, followed by CTX-M-14 (36.4%) and CTX-M-15 (27.9%) with IncF plasmids. BlaKPC-2 was carried by FIIAs plasmids. Epidemiology typing revealed 8 K. pneumoniae ST types—ST15(8/20), ST17(4/20), ST37(2/20) and 9 E. coli ST types—ST131 (30.9%, 21/68), ST38 (8/68), ST95(7/68) and ST316(7/68). All ST131 isolates but one was from the highly virulent epidemic clone O25bST131. This is the first report in Bulgaria about ESBL/carbapenemase faecal carriage. We observed high ESBL/carbapenemases prevalence. A predominant number of isolates were members of highly epidemic and virulent PanEuropean clones ST15 K. pneumoniae and O25bST131 E. coli. High antibiotics usage during the COVID pandemic will worsen the situation. Routine screenings and strict infection control measures should be widely implemented.


2020 ◽  
Vol 89 (3) ◽  
pp. 217-223
Author(s):  
Jan Vašek ◽  
Jonáš Vaňhara ◽  
Monika Dolejská ◽  
Martina Masaříková ◽  
Alois Čížek ◽  
...  

The aim of the present study was to monitor the presence of extended-spectrum beta-lactamase (ESBL) producing E. coli on farm A with the history of previous use of ceftiofur in suckling pigletsand to analyse the risk factors of selection and dissemination of ESBL producers in the production herd. In the year of 2014, a total of 411 samples (rectal swabs or faeces)from pigs of various age categories (sows, gilts and suckling piglets) were collected. The sampling was performed more than 24 months after the ban of ceftiofur on the farm.The sows and gilts were sampled repeatedly before and after farrowing. All collected samples were directly cultivated on MacConkey agar (MCA) containing cefotaxime (2 mg/l) and obtained sub-cultures were tested for ESBL production by double disc synergy test. According to our results, all gilts were negative for ESBL-producing E. coli in the introduction period, however, the excretion of ESBL-producing E. coli was observed before and after delivery. Most of the new-born piglets from positive sows and gilts shed ESBL-producing E. coli early after birth. All tested ESBL-producing isolates were resistant to multiple antimicrobials, suggesting that antibiotics from other groups used for therapy co-select for ESBL producers in pigs on the studied farm. Intestinal colonization of lactating sows and their offspring as well as survival of ESBL-producing E. coli in the farm environment should be recognised as important risk factors of circulation and long-time persistence of ESBL producers in the herd.


2019 ◽  
Vol 15 (3) ◽  
pp. 211-217
Author(s):  
Surya Narayan Mahaseth ◽  
Raj Kumari Sanjana ◽  
Brajesh Kumar Jha ◽  
Khilasa Pokharel

Background: Urinary tract infection (UTI) is one of the major health problems in Nepal. Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) are two important bacteria associated with UTI. This study was designed to assess the prevalence of extended spectrum beta-lactamase (ESBL) producing E. coli and K. pneumoniae in urinary isolates at College of Medical Sciences and Teaching Hospital, Bharatpur, Chitwan, Nepal. Methods: We aseptically received 5564 mid-stream urine samples of suspected UTI patients from September 2016 to August 2018. The randomly collected 5564 urine samples were processed by standard Microbiological guidelines as recommended by Clinical and Laboratory Standards Institute (CLSI). All isolates including E. coli and K. pneumoniae were identified using the specific biochemical and sugar fermentation tests. Antibiotic sensitivity test was performed for all the isolates against all commonly used antibiotics by modified Kirby-Bauer disk diffusion method and interpreted following CLSI guidelines. First performed initial screening method then confirmed for ESBL production by phenotypic confirmatory disc diffusion test (PCDDT). Results: Out of 5,564 urine specimens investigated, E. coli was isolated in 1219 (63.99%) and K. pneumoniae in 223 (11.70%) cases. Initial screening revealed 615 (50.45%) isolates of E. coli and 127 (56.95%) K. pneumoniae to be resistant. Further testing by PCDDT method confirmed 102 (16.58%) E. coli and 25 (19.68%) K. pneumoniae isolates to be ESBL producers. These ESBL producers’ uropathogens revealed high degree of resistance to cephalosporins (100%) and quinolones (52%-92%) group of antibiotics. Conclusions: In our study the prevalence of ESBL producing K. pneumoniae was found to be 19.68%, those of E. coli was to be 16.58% by PCDDT. In this study, all ESBL producing K. pneumoniae isolates were sensitive (100%) to meropenem and E. coli showed 98.04% sensitive to meropenem. Hence, for the treatment of these ESBL infections, currently, carbapenems are the recommended drug of choice.


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