Prevalence of Extended-Spectrum β-Lactamase-Production among Clinical Gram-Negative Bacilli Isolates in A Tertiary Hospital, Southwest Nigeria

Extended spectrum β-lactamases (ESBLs) have emerged as important mechanisms of antimicrobial resistance to beta-lactam drugs in hospitals, thereby limiting the available treatment options for infections caused by these microorganisms. The objectives of this study were to determine the prevalence of ESBL production among clinical isolates of Gram-negative bacilli from Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Nigeria as part of the global efforts to provide information for the containment of the spread of antimicrobial resistance. This is a laboratory-based study of 186 consecutive non-duplicate Gram-negative bacilli (GNB) isolated from diverse clinical samples at the Microbiology laboratory of the hospital after standard aerobic cultures. The isolates were identified by conventional biochemical test scheme and commercial GNB API 20E identification kit. Antimicrobial susceptibility testing (AST) of each isolate was determined by the disk diffusion technique. Isolates presumptively identified as ESBL producers from the AST assay were confirmed by the combination disks method. Escherichia coli (n=43; 23.1%) predominated among the organisms, followed by Enterobacter spp (n=23; 12.4%) and Klebsiella spp (n=22; 11.8%). Most of the organisms (95.7%) were resistant to tetracycline and ampicillin, but the least resistance (4.3%) was exhibited to imipenem. Fifty-eight (31.2%) isolates were ESBL producers with a preponderance of E. coli (n=15; 25.9%). The ESBL producers were commonly isolated from urine samples (n=31; 53.4%), and were significantly more resistant to levofloxacin, ciprofloxacin, nalidixic acid, gentamicin, augmentin, cefepime, ceftriaxone, ceftazidime and cefotaxime (p<0.05). This study has highlighted the therapeutic implications of the presence of EBSL in clinical isolates which was depicted by their multi-drug resistance phenotypes, thus emphasizing the need for continuous surveillance of bacterial resistance, institution of antimicrobial resistance stewardship in our hospital and regular screening of ESBLs in clinical isolates to prevent treatment failure.

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Prevalence of multidrug-resistant and extended-spectrum beta-lactamase producing Gram-negative isolates from clinical samples in a tertiary care hospital of Nepal

Tropical Medicine and Health ◽

10.1186/s41182-021-00313-3 ◽

2021 ◽

Vol 49 (1) ◽

Author(s):

Aryatara Shilpakar ◽

Mehraj Ansari ◽

Kul Raj Rai ◽

Ganesh Rai ◽

Shiba Kumar Rai

Keyword(s):

Tertiary Care Hospital ◽

Tertiary Care ◽

Polymyxin B ◽

Multidrug Resistant ◽

Confirmatory Test ◽

Clinical Samples ◽

Care Hospital ◽

Gram Negative ◽

Extended Spectrum ◽

Esbl Producers

Abstract Background The existence of multidrug-resistant organisms, including extended-spectrum beta-lactamases (ESBLs), is on rise across the globe and is becoming a severe problem. Knowledge of the prevalence and antibiogram profile of such isolates is essential to develop an appropriate treatment methodology. This study aimed to study the prevalence of Gram-negative isolates exhibiting ESBL at a tertiary care hospital and study their antibiogram profile. Methods A cross-sectional study was conducted at Shahid Gangalal National Heart Centre, Kathmandu, Nepal, from June 2018 to November 2018. A total of 770 clinical samples were collected and identified using the conventional biochemical tests following the Clinical and Laboratory Standard Institute (CLSI) guidelines. Antimicrobial susceptibility testing (AST) was performed using the standardized Kirby-Bauer disk diffusion method. The screening test for ESBL producers was performed as recommended by the CLSI and the confirmatory test was performed phenotypically using the E-test. Results Out of the 92 isolates, 84 (91.3%) were multidrug-resistant, and 47 (51.1%) were found to be potential ESBL producers. Of these, 16 isolates were confirmed ESBL producers by the E-test. Escherichia coli and Klebsiella pneumoniae were the predominant isolates and were also the major ESBL producers. Besides polymyxin B (100% sensitive), meropenem and imipenem showed high efficacy against the ESBL producers. Conclusion Multidrug resistance was very high; however, ESBL production was low. Polymyxin B and carbapenems are the choice of drugs against ESBL producers but should be used only as the last line drugs.

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Antimicrobial Resistance Patterns in Clinical Isolates of Enterobacteriaceae from a Tertiary Care Hospital, Kathmandu, Nepal

Nepalese Medical Journal ◽

10.3126/nmj.v1i2.21578 ◽

2018 ◽

Vol 1 (2) ◽

pp. 74-78

Author(s):

Ram Prasad Adhikari ◽

Subha Shrestha ◽

Junu Richhinbung Rai ◽

Ritu Amatya

Keyword(s):

Multidrug Resistance ◽

Antimicrobial Resistance ◽

College Teaching ◽

Clinical Isolates ◽

High Rate ◽

Clinical Samples ◽

Care Hospital ◽

Beta Lactamases ◽

Extended Spectrum ◽

Extended Spectrum Beta Lactamases

Introduction: Multidrug resistance among Enterobacteriaceae is in increasing trend these days. The objective of this study was to determine the antibiogram of clinical isolates of Enterobacteriaceae with special reference to multidrug resistance and extended spectrum beta-lactamases production.Materials and Methods: A descriptive cross sectional study was conducted over a period of six months (February -July, 2017) in the microbiology laboratory of Nepal Medical College Teaching Hospital, Kathmandu, Nepal. A total of 936 bacterial isolates of Enterobacteriaceae from clinical specimens were processed for antimicrobial susceptibility testing and screened for multidrug resistance. ESBL production was detected among potential isolates by combination disk diffusion test.Results: The rate of multidrug resistance and extended spectrum beta-lactamases production was 54.2% and 23.8% respectively. Of the total ESBL producers 92.4% were multidrug resistance. The rate of multidrug resistance and extended spectrum beta-lactamases production were higher in organisms isolated from clinical samples collected from inpatients. High rate of multidrug resistance and extended spectrum beta-lactamases production was seen in E. coli (54.4% & 27.7%), Klebsiella spp. (67.1% & 28.2%) and Citrobacter spp. (70.3% & 10.9%). The antimicrobial resistance rate was highest against ampicillin (76.7%) followed by cefixime (54. 0%), ceftazidime (51.5%), ceftriaxone (51.0%), cotrimoxazole (48.7%), ciprofloxacin (43.9%) and ofloxacin (41.1%).Conclusions: Multidrug resistance is common among Enterobacteriaceae. These bacteria have high rate of resistance against commonly used groups of antibiotics like cephalosporins and quinolones. Continuous monitoring, surveillance of antimicrobial resistance, proper infection control and practices are important to combat with these issues.

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Antimicrobial Resistance and Prevalence of Extended Spectrum β-Lactamase Among Clinical Isolates of Gram-Negative Bacteria in Riyadh

Journal of Chemotherapy ◽

10.1179/joc.1995.7.6.509 ◽

1995 ◽

Vol 7 (6) ◽

pp. 509-514 ◽

Cited By ~ 5

Author(s):

T. El-Karsh ◽

A.F. Tawfik ◽

F. AL-Shammary ◽

S. AL-Salah ◽

A.M. Kambal ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Clinical Isolates ◽

Gram Negative Bacteria ◽

Gram Negative ◽

Extended Spectrum

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Detection of TEM and CTX-M Genes in Escherichia coli Isolated from Clinical Specimens at Tertiary Care Heart Hospital, Kathmandu, Nepal

Diseases ◽

10.3390/diseases9010015 ◽

2021 ◽

Vol 9 (1) ◽

pp. 15

Author(s):

Ram Shankar Prasad Sah ◽

Binod Dhungel ◽

Binod Kumar Yadav ◽

Nabaraj Adhikari ◽

Upendra Thapa Shrestha ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Clinical Isolates ◽

Diffusion Method ◽

Gram Negative Bacteria ◽

Bacterial Isolates ◽

Disk Diffusion Method ◽

Gram Negative ◽

Clinical Specimens ◽

E Coli ◽

Esbl Producers

Background: Antimicrobial resistance (AMR) among Gram-negative pathogens, predominantly ESBL-producing clinical isolates, are increasing worldwide. The main aim of this study was to determine the prevalence of ESBL-producing clinical isolates, their antibiogram, and the frequency of ESBL genes (blaTEM and blaCTX-M) in the clinical samples from patients. Methods: A total of 1065 clinical specimens from patients suspected of heart infections were collected between February and August 2019. Bacterial isolates were identified on colony morphology and biochemical properties. Thus, obtained clinical isolates were screened for antimicrobial susceptibility testing (AST) using modified Kirby–Bauer disk diffusion method, while ESBL producers were identified by using a combination disk diffusion method. ESBL positive isolates were further assessed using conventional polymerase chain reaction (PCR) to detect the ESBL genes blaTEM and blaCTX-M. Results: Out of 1065 clinical specimens, 17.8% (190/1065) showed bacterial growth. Among 190 bacterial isolates, 57.4% (109/190) were Gram-negative bacteria. Among 109 Gram-negative bacteria, 40.3% (44/109) were E. coli, and 30.2% (33/109) were K. pneumoniae. In AST, 57.7% (n = 63) Gram-negative bacterial isolates were resistant to ampicillin and 47.7% (n = 52) were resistant to nalidixic acid. Over half of the isolates (51.3%; 56/109) were multidrug resistant (MDR). Of 44 E. coli, 27.3% (12/44) were ESBL producers. Among ESBL producer E. coli isolates, 58.4% (7/12) tested positive for the blaCTX-M gene and 41.6% (5/12) tested positive for the blaTEM gene. Conclusion: Half of the Gram-negative bacteria in our study were MDR. Routine identification of an infectious agent followed by AST is critical to optimize the treatment and prevent antimicrobial resistance.

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Identification and Antibiotic Susceptibility Testing of ESBL producing Klebsiella strains by Phenotypic methods

RADS Journal of Biological Research & Applied Sciences ◽

10.37962/jbas.v9i1.147 ◽

2018 ◽

Vol 9 (1) ◽

pp. 8-13

Author(s):

Malik Taqdees ◽

Asma Naim ◽

Asma Saeed

Keyword(s):

Diffusion Method ◽

Multi Drug Resistance ◽

Beta Lactamase ◽

Disk Diffusion Method ◽

Klebsiella Species ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Klebsiella Spp ◽

Esbl Producers

Multi drug resistance has now become a worldwide therapeutic challenge due to the widespread use of broad spectrum antibiotics. Klebsiella species have significant importance in clinical field as they cause various infections in human and are considered as potential pathogens that express antibiotic resistance through their strong enzymatic activity. Extended spectrum beta lactamases (ESBLs) are plasmid mediated enzymes produced mostly because of mutation and few other factors. These enzymes confer resistance against various Î²-lactam drugs including cephalosporins and monobactams. Among the genus Klebsiella, ESBLs are highly prevalent in K. pneumoniae followed by K. oxytoca. This study was conducted in Pakistan to assess the distribution of ESBL producers among Klebsiella spp., an important member of the family Enterobacteriaceae. From January 2010 to January 2012, a total of 236 gram-negative isolates were collected from different renowned microbiological laboratories. Out of the 236 gram-negative isolates, 125 were found as Klebsiella spp. by using standard microbiological techniques. Antimicrobial susceptibility profiling of these strains was performed by using Kirby Bauer disk diffusion method. Phenotypic detection of the production of extended spectrum beta lactamase enzyme was performed using double disc synergy method and combination disc method. It has been identified that Klebsiella strains are highly resistant against Amoxicillin, Tetracycline, Nalidixic Acid, Cephradine, Gentamicin, co-amoxyclav with the percentage of 100%, 86%, 86%, 82%, 82% and 80% respectively. The most effective antibiotics for Klebsiella spp. were found to be Amikacin, Meropenem and Piperacillin-tazobactam, with highest sensitivities of 96%, 94% and 91%. Phenotypic detection of Extended spectrum beta lactamase production by double disc synergy test was able to identify 28% ESBL producers among Klebsiella isolates whereas 64% were detected by combination disc test.

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A Tale of β-Lactamases and Multidrug Resistance in Pseudomonas aeruginosa Isolated from Inpatients in a University Hospital

Journal of Institute of Medicine ◽

10.3126/jiom.v43i1.37467 ◽

2021 ◽

Vol 43 (1) ◽

pp. 19-24

Author(s):

Santosh K Yadav ◽

Sangita Sharma ◽

Shyam K Mishra ◽

Jeevan B Sherchand

Keyword(s):

Pseudomonas Aeruginosa ◽

Antimicrobial Resistance ◽

Critical Role ◽

Modern Medicine ◽

Hospitalized Patients ◽

University Teaching ◽

University Hospital ◽

Clinical Samples ◽

Extended Spectrum ◽

Tract Infections

Introduction In this era of modern medicine, antimicrobial resistance can be regarded as a major health calamity. The emergence of multidrug-resistant (MDR) Pseudomonas aeruginosa strains poses therapeutic challenges and lead to treatment failure in hospitalized patients. This study was conducted to determine various types of β-lactamases among MDR P. aeruginosa isolates recovered from hospitalized patients. MethodsThis study was conducted at Tribhuvan University Teaching Hospital, Maharajgunj, Nepal. The clinical samples collected from inpatients were processed for detection of P. aeruginosa isolates and antibiotic susceptibility profile was determined. The MDR strains were identified and ceftazidime-resistant isolates were subjected for detection of extended-spectrum-β-lactamase (ESBL), metallo-β-lactamase (MBL), and Klebsiella pneumoniae carbapenemase (KPC). ResultsA total of 161 P. aeruginosa isolates were recovered during the study period encompassing 73.3% (n=118) MDR isolates. The MDR isolates included 50.0% (n=59) from lower respiratory tract infections; and 39.8% (n=47) were from the intensive care unit patients. The MDR isolates showed a high resistance profile towards piperacillin, cephalosporins, and fluoroquinolones (>85%). Resistance to carbapenems and aminoglycosides were up to 80% and 60% respectively. Extended spectrum-β-lactamase, MBL, and KPC mediated resistance were seen in 34.7%, 43.6%, and 14.4% MDR isolates, respectively. ConclusionMultidrug resistance as well as resistance mediated by β-lactamases production were high among P. aeruginosa isolates. Therefore, early detection of antimicrobial resistance and rational use of antibiotics play a critical role to fight against this MDR pathogen.

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Extended Spectrum Beta-Lactamases Producers among Gram-negative Bacteria from Clinical and Environmental Sources in Two Tertiary Hospitals in Makurdi, Nigeria

Microbiology Research Journal International ◽

10.9734/mrji/2020/v30i530220 ◽

2020 ◽

pp. 42-53

Author(s):

Florence Bose Omoregbe ◽

Obasola Ezekiel Fagade

Keyword(s):

Clinical Isolates ◽

Diffusion Method ◽

Clinical Samples ◽

Multiple Drug ◽

Gram Negative Bacteria ◽

Disc Diffusion ◽

Gram Negative ◽

Beta Lactamases ◽

Extended Spectrum ◽

Extended Spectrum Beta Lactamases

Extended Spectrum Beta-Lactamases (ESBLs) enzymes are produced by many Gram-negative bacteria to nearly all antibiotics. Clinical bacteria isolates were obtained from various clinical samples from the two healthcare facilities. Sewer wastewater and sediments were also collected from both hospitals using standard sampling techniques and bacteria isolated using pour plate technique. Multiple drug resistant patterns were determined using disc diffusion technique. Antibiotics sensitivity of the isolates was verified using disc diffusion method. Detection of ESBL producing bacteria was done using double disc synergy test. Data obtained were analysed using descriptive statistics. Clinical bacteria (403) were obtained, out of which 299 were confirmed Gram-negative, 218 from Federal Medical Centre (FMC) and 81 from Benue State Teaching Hospital (BSUTH). Thirty-nine Gram-negative bacteria were also isolated from the environmental samples. The ESBL producers in clinical isolates were 69 (FMC) and 42 (BSUTH) while in environmental isolates they were eight and four respectively. Out of a total of 338 environmental and clinical isolates from the two hospitals, 216 shown resistance/intermediate resistance to Ceftazidime. Of these 216, 123(36.39%) were positive for ESBLs production.- space From FMC, ESBLs producing bacteria are highly sensitive to imipenem with sensitivity frequency of 62 while they were highly resistant to ceftazidime with a frequency of 52. Age groups 0-5 had the highest percentage distribution of 21.43% and 10.14% from BSUTH and FMC respectively. ESBLs producing bacteria showed multidrug resistance.

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High Prevalence of Multi-Drug Resistance and Extended Spectrum Beta Lactamase Production in Non-Fermenting Gram-Negative Bacilli in Ethiopia

Infectious Diseases Research and Treatment ◽

10.1177/1178633719884951 ◽

2019 ◽

Vol 12 ◽

pp. 117863371988495 ◽

Cited By ~ 1

Author(s):

Adane Bitew

Keyword(s):

Drug Resistance ◽

Bacterial Species ◽

Multi Drug Resistance ◽

Clinical Samples ◽

Beta Lactamase ◽

Drug Resistant ◽

Acinetobacter Species ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum

Background: Emergence of resistance to multiple antimicrobial agents in Non-Fermenting Gram-Negative Bacilli is a major problem to public health, as it limits drug treatment options against infections. The aim of this study was to determine the prevalence of multi-drug resistance and extended spectrum beta lactamase production in Non-Fermenting Gram-Negative Bacilli. Materials and methods: Different clinical samples were collected and processed following standard procedures. Each sample was then inoculated onto culture media. Identification, drug susceptibility testing, and extended spectrum beta lactamase production of the isolates were carried out by using the VITEK 2 compact system. Results: Among 996 clinical samples, 135 samples yielded Non-Fermenting Gram-Negative Bacilli of which Pseudomonas and Acinetobacter species were the commonest isolates. The overall drug resistance rates of Non-Fermenting Gram-Negative Bacilli were above 80% against ampicillin (89.6%), cefuroxime axetil (88.9%), nitrofurantoin (85.9%), cefalotin (84.4%), cefoxitin (83.7%), cefazolin (83.0%), and cefuroxime (83.0%). Tobramycin with a resistance rate of 19.3% was the most active antimicrobial agent. Out of 135 isolates, 81.5% were multi-drug resistant of which 13.3% were extensively drug resistant and 10.4% were pandrug resistant. Extended spectrum beta lactamase production was detected in 48.9% of the isolates. Conclusions: The spectrum of bacterial species isolated was diverse. The isolates demonstrated high level of drug resistance in different classes of antibiotics. The magnitude of multi-drug resistance and the level of extended spectrum beta lactamase production were high. Hence, further studies on multi-drug resistant and extended spectrum beta lactamase producing Non-Fermenting Gram-Negative Bacilli both in the community and in hospital setting are essential.

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A One Health Comparative Assessment of Antimicrobial Resistance in Generic and Extended-Spectrum Cephalosporin-Resistant Escherichia coli from Beef Production, Sewage and Clinical Settings

Microorganisms ◽

10.3390/microorganisms8060885 ◽

2020 ◽

Vol 8 (6) ◽

pp. 885 ◽

Cited By ~ 1

Author(s):

Emelia H. Adator ◽

Claudia Narvaez-Bravo ◽

Rahat Zaheer ◽

Shaun R. Cook ◽

Lisa Tymensen ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

One Health ◽

Hot Spot ◽

Municipal Sewage ◽

Multi Drug Resistance ◽

Beef Production ◽

E Coli ◽

Beef Processing ◽

Extended Spectrum

This study aimed to compare antimicrobial resistance (AMR) in extended-spectrum cephalosporin-resistant and generic Escherichia coli from a One Health continuum of the beef production system in Alberta, Canada. A total of 705 extended-spectrum cephalosporin-resistant E. coli (ESCr) were obtained from: cattle feces (CFeces, n = 382), catch basins (CBasins, n = 137), surrounding streams (SStreams, n = 59), beef processing plants (BProcessing, n = 4), municipal sewage (MSewage; n = 98) and human clinical specimens (CHumans, n = 25). Generic isolates (663) included: CFeces (n = 142), CBasins (n = 185), SStreams (n = 81), BProcessing (n = 159) and MSewage (n = 96). All isolates were screened for antimicrobial susceptibility to 9 antimicrobials and two clavulanic acid combinations. In ESCr, oxytetracycline (87.7%), ampicillin (84.4%) and streptomycin (73.8%) resistance phenotypes were the most common, with source influencing AMR prevalence (p < 0.001). In generic E. coli, oxytetracycline (51.1%), streptomycin (22.6%), ampicillin (22.5%) and sulfisoxazole (14.3%) resistance were most common. Overall, 88.8% of ESCr, and 26.7% of generic isolates exhibited multi-drug resistance (MDR). MDR in ESCr was high from all sources: CFeces (97.1%), MSewage (96.9%), CHumans (96%), BProcessing (100%), CBasins (70.5%) and SStreams (61.4%). MDR in generic E. coli was lower with CFeces (45.1%), CBasins (34.6%), SStreams (23.5%), MSewage (13.6%) and BProcessing (10.7%). ESBL phenotypes were confirmed in 24.7% (n = 174) ESCr and 0.6% of generic E. coli. Prevalence of bla genes in ESCr were blaCTXM (30.1%), blaCTXM-1 (21.6%), blaTEM (20%), blaCTXM-9 (7.9%), blaOXA (3.0%), blaCTXM-2 (6.4%), blaSHV (1.4%) and AmpC β-lactamase blaCMY (81.3%). The lower AMR in ESCr from SStreams and BProcessing and higher AMR in CHumans and CFeces likely reflects antimicrobial use in these environments. Although MDR levels were higher in ESCr as compared to generic E. coli, AMR to the same antimicrobials ranked high in both ESCr and generic E. coli sub-populations. This suggests that both sub-populations reflect similar AMR trends and are equally useful for AMR surveillance. Considering that MDR ESCr MSewage isolates were obtained without enrichment, while those from CFeces were obtained with enrichment, MSewage may serve as a hot spot for MDR emergence and dissemination.

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Detection of Extended-Spectrum β-Lactamases in Clinical Isolates of Pseudomonas aeruginosa

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01511-05 ◽

2006 ◽

Vol 50 (9) ◽

pp. 2990-2995 ◽

Cited By ~ 70

Author(s):

Xiaofei Jiang ◽

Zhe Zhang ◽

Min Li ◽

Danqiu Zhou ◽

Feiyi Ruan ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Multidrug Resistant ◽

Clinical Isolates ◽

Screening Methods ◽

Extended Spectrum ◽

Pcr Product ◽

Amoxicillin Clavulanate ◽

Synergy Test ◽

Esbl Producers ◽

Disk Test

ABSTRACT With the occurrence of extended-spectrum β-lactamases (ESBLs) in Pseudomonas aeruginosa being increasingly reported worldwide, there is a need for a reliable test to detect ESBLs in clinical isolates of P. aeruginosa. In our study, a total of 75 clinical isolates of P. aeruginosa were studied. Nitrocefin tests were performed to detect the β-lactamase enzyme; isoelectric focusing electrophoresis, PCR, and PCR product sequencing were designed to further characterize the contained ESBLs. Various ESBL-screening methods were designed to compare the reliabilities of detecting ESBLs in clinical isolates of P. aeruginosa whose β-lactamases were well characterized. Thirty-four of 36 multidrug-resistant P. aeruginosa clinical isolates were positive for ESBLs. bla VEB-3 was the most prevalent ESBL gene in P. aeruginosa in our study. Among the total of 34 isolates that were considered ESBL producers, 20 strains were positive using conventional combined disk tests and 10 strains were positive using a conventional double-disk synergy test (DDST) with amoxicillin-clavulanate, expanded-spectrum cephalosporins, aztreonam, and cefepime. Modifications of the combined disk test and DDST, which consisted of shorter distances between disks (20 mm instead of 30 mm) and the use of three different plates that contained cloxacillin (200 μg/ml) alone, Phe-Arg β-naphthylamide dihydrochloride (MC-207,110; 20 μg/ml) alone, and both cloxacillin (200 μg/ml) and MC-207,110 (20 μg/ml) increased the sensitivity of the tests to 78.8%, 91.18%, 85.29%, and 97.06%.

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