scholarly journals Phenotypic characterization of beta-lactamases producing Gram-negative bacteria in a tertiary hospital, Nepal

2019 ◽  
Vol 7 (1) ◽  
pp. 74-81
Author(s):  
Elina Maharjan ◽  
Pooja Shakya ◽  
Balkrishna Bhattachan ◽  
Bharat Prasad Baral ◽  
Dhiraj Shrestha

Infections caused by beta-lactamases producing Gram-negative bacteria are increasing, thus posing a challenge to the management of such infections. The surveillance data of such bacteria is limited in Nepal so this study aimed to detect the beta-lactamase producing Gram-negative bacteria in a tertiary setting. A total of 604 clinical samples, including urine, blood, sputum and body fluids, were cultured and identified by the routine standard laboratory protocols. Antibiotic susceptibility testing was done by Kirby Bauer disc diffusion method following Clinical and Laboratory Standard Institute guidelines (2014). Extended-spectrum beta-lactamases (ESBL) producers were identified by combined disk method and metallo-beta-lactamases (MBL) producers were identified by Imipenem- EDTA combined disk method. Out of 604 samples, 282 (46.7%) samples showed significant growth, of which 229 (81.2%) were Gram-negative bacteria. Of 229 Gram-negative bacteria, 200 (87.3%) were multidrug resistant, 67 (29.3%) were ESBL producers and 16 (7.0%) were MBL producers. Klebsiella pneumoniae were among higher ESBL producers and Pseudomonas aeruginosa were among higher MBL producers. The findings suggest higher antibacterial resistance among Gram-negative bacteria with the added burden of beta-lactamase production. Imipenem was effective against 125 of 229 Gram-negative bacteria tested. Thus, imipenem can be the drug of choice for empirical management. The higher multidrug resistance and higher beta-lactamases production among Gram-negative bacteria warrant the continuous monitoring, surveillance, early detection, and infection control practices of such bacteria

KYAMC Journal ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 171-175
Author(s):  
Tania Rahman ◽  
Momtaz Begum ◽  
Sharmeen Sultana ◽  
SM Shamsuzzaman

Background: In recent years, Extended-spectrum beta-lactamase (ESBL) producing microorganisms have complicated treatment of infections due to resistance of ESBL producing strains to a wide range of antimicrobials. Objective: Target of this study was to determine the prevalence of ESBL producing gramnegative bacteria in neonatal sepsis cases and to reveal the antimicrobial susceptibility pattern of those isolated ESBL producers. Materials and Methods: This cross sectional study was carried out in Dhaka Medical College Hospital (DMCH) over a period of 12 months from January to December in 2016. Following isolation and identification of gram-negative bacteria from blood samples of suspected septicemic neonates, antimicrobial susceptibility test was performed by Kirby Bauer disk-diffusion method and ESBL producers were detected by Double Disk Synergy (DDS) test. Results: Among 52 Gram-negative bacteria isolated from 106 blood samples, 34.61% ESBL producers were detected and Enterobacter spp. (45%) was predominant followed by Klebsiella pneumoniae (33.33%). None of the ESBL producers was resistant to colistin and tigecycline. All ESBL producing Acinetobacter baumannii, 77.78% and 66.67% of ESBL producing Enterobacter spp and Klebsiella spp. respectively showed resistance to meropenem. All ESBL producers were resistant to piperacillintazobactam. Conclusion: Appropriate measures should be taken to prevent the spread of ESBL producing strains by combining strategies for infection prevention, control and rational use of antibiotics. KYAMC Journal Vol. 11, No.-4, January 2021, Page 171-175


Author(s):  
Kavi Aniis ◽  
Rajamanikandan Kcp ◽  
Arvind Prasanth D

<p>ABSTRACT<br />Objective: Beta-lactams are the group of antibiotics that contain a ring called as “beta-lactam ring,” which is responsible for the antibacterial activity.<br />The presence of resistance among Gram-negative organisms is due to the production of beta-lactamases enzymes that hydrolysis the beta-lactam ring<br />thereby conferring resistance to the organism. This study is undertaken to determine the prevalence of extended-spectrum beta-lactamase (ESBL)<br />producing Gram-negative organism from clinical samples.<br />Methods: A total of 112 clinical samples were taken for this study. The combined disc synergistic test (CDST) was used for the phenotypic detection<br />of ESBL producers from the clinical samples. The genotypic identification of ESBL producers was carried out by alkaline lysis method by isolation of<br />plasmid DNA.<br />Result: A total of 87 bacterial isolates were isolated and identified. Among them, Klebsiella (41%) was the predominant organism followed by<br />Escherichia coli (33%), Proteus (10%), Pseudomonas (10%), and Serratia (6%). Among the various bacterial isolates, Klebsiella showed a higher<br />percentage of resistance. The CDST showed that 8 isolates of Klebsiella, 3 isolates of E. coli, and 1 isolate of Pseudomonas were found to be ESBL<br />producers. The genotypic confirmation showed that the two bacterial isolates, namely, Klebsiella and E. coli were found to possess temoniera (TEM)<br />gene which was the 400-500 bp conferring resistance to the antibiotics.<br />Conclusion: The results of this study suggest that early detection of ESBL producing Gram-negative organism is a very important step in planning the<br />therapy of patient in Hospitals. CDST continues to be a good indicator in the detection of ESBL producers.<br />Keywords: Beta-lactamases, Gram-negative bacilli, Extended-spectrum beta-lactamase, Resistance, Combined disc synergistic test.</p><p> </p>


2020 ◽  
Vol 14 (3) ◽  
pp. 2027-2032
Author(s):  
Mita D. Wadekar ◽  
J.V. Sathish ◽  
C. Pooja ◽  
S. Jayashree

Resistance to beta lactam antibiotics is the most common cause for beta-lactamase production. Increasing number of extended spectrum beta-lactamase (ESBL) producers has reduced the treatment options which resulted in emergence of multidrug resistant strains, treatment failure and hence increased mortality. To detect phenotypically, ESBL producers in Gram negative isolates from different samples and to know their susceptibility pattern. A retrospective study of Gram negative isolates was conducted. Total of 521 isolates were isolated from various samples. They were processed and identified by standard procedures. The antibiotic susceptibility testing was performed by Kirby- Bauer disc diffusion method using CLSI guidelines. ESBL was detected by combination disk test. A total of 521 Gram negative isolates were isolated which included E. coli, Klebsiella pneumoniae, Citrobacter spp., Enterobacter spp., Proteus spp. and Acinetobacter spp. Pseudomonas aeruginosa. Of 521 isolates tested, ESBL was detected in 329 (63.1%) isolates. These isolates showed maximum susceptibility to piperacillin- tazobactam (86%) followed by imipenem (78.4%), amikacin (63.5%), cotrimoxazole (54.4%), ciprofloxacin (51%), amoxi-clav (44.9%), cefepime (44.1%), gentamicin (38.9%), cefoxitin (34.9%) and ampicillin (19.1%). ESBL producers which are resistant to beta lactam antibiotics have become a major problem. Detection of these beta-lactamase enzymes by simple disk method and its reporting will help clinicians in prescribing proper antibiotics.


Author(s):  
Florence Bose Omoregbe ◽  
Obasola Ezekiel Fagade

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.


Author(s):  
Jashmi Chandraker ◽  
Ashish Saraf ◽  
Sachin Chandraker ◽  
Kanupriya Dubey

The rapid spread of acquired metallo-beta-lactamases (MBLs) among major Gram-negative pathogens is an emerging threat and a matter of particular concern worldwide. Carbapenems are among the  few useful antibiotics against gram negative bacteria particularly those  with extended spectrum beta-lactamase. Resistant to carbapenem is mediated by loss of outer membrane proteins and production of beta lactamase that is capable of hydrolyzing carbapenems.During this study period, 80 different clinical samples were collected from various hospitals of Raipur. All clinical samples were processed according to standard microbiological methods. Isolated GNRs were subjected to susceptibility testing against antibiotics by disc diffusion method according to the Clinical and Laboratory Standards Institute (CLSI) guidelines.  Carbapenem-resistant isolates were subjected to the detection of MBL production by different methods.  Out of 80 samples, 28 (35%) samples showed significant growth. MBL production was observed in 20 samples.


2016 ◽  
Vol 6 (2) ◽  
pp. 94-99
Author(s):  
Amir Ibrahimagić ◽  
Emina Idrizović ◽  
Edita Divjan ◽  
Biljana Klimenta

Introduction: Infections due to extended-spectrum beta-lactamase (ESBL)-producing isolates in patients are hard to treat and cause high morbidity and mortality. ESBL-producing bacteria have been increasingly detected in environmental samples in different countries since 2002, and have gained considerable attention worldwide.Methods: Antibiotic susceptibility of all isolates was determined using the disk diffusion method. The production of ESBLs was determined by the double-disk synergy test.Results: Among the outpatient clinical samples, out of 2857 Gram-negative bacteria, 184 (6.5%) ESBL-producing bacteria were isolated. In this group, 143 (77.7%) were from urine samples, 26 (14.1%) from surgical wounds, 6 (3.3%) from umbilical swabs, and 9 (4.9%) from other patients sites (upper respiratory tract, cannula, eyes, genital swabs). Escherichia coli was isolated in 62 (33.7%), and Klebsiella spp. in 50 (27.8%) cases. Among the environmental samples, out of 381 Gram-negative bacteria, 52 (13.6%) were ESBL-producing isolates. In this group, 37 (71.2%) were sampled from water, 7 (13.5%) from food, and 8 (15.4%) from environmental surfaces. The most prevalent ESBL-producing bacteria isolated from the environmental samples were E. coli (isolated from 26 samples), Klebsiella spp. (10), non-fermenters (9), and other bacteria isolated from 7 samples. The clinical outpatient ESBL-producing isolates showed resistance to all cephalosporins, ranging from 25% (cefepime) to 100% (cefuroxime). The environmental ESBL-producing isolates showed resistance to cefuroxime, aztreonam, cefpodoxime, amoxicillin/clavulanate, and cefoxitin in the range of 65-100%.Conclusions: Prevalence of antibiotic resistance of ESBL-producing strains is high and requires routine detection of ESBL-producing isolates in the laboratories, designing of appropriate antibiotic prescribing policies and control of the risk factors.


Author(s):  
Ganiyat Shitta ◽  
Olufunmilola Makanjuola ◽  
Olusolabomi Adefioye ◽  
Olugbenga Adekunle Olowe

Background: Extended Spectrum Beta Lactamase (ESBL) production in gram negative bacteria confers multiple antibiotic resistance, adversely affecting antimicrobial therapy in infected individuals. ESBLs result from mutations in β-lactamases encoded mainly by the bla TEM,bla SHVand bla CTX-Mgenes. The prevalence of ESBL producing bacteria has been on the increase globally especially its upsurge among isolates from community-acquired infections. Aim: To determine ESBL prevalence and identify ESBL genes among clinical isolates in Osun State, Nigeria. Material and Methods: A cross-sectional study was carried out from August 2016 –July 2017 in Osun State, Nigeria. Three hundred and sixty Gram negative bacteria recovered from clinical samples obtained from both community and healthcare associated infections were tested. They included147 Escherichia coli(40.8%), 116 Klebsiella spp(32.2%), 44 Pseudomo-nas aeruginosa(12.2%) and23 Proteus vulgaris (6.4%) isolates. Others were Acinetobacter baumannii, Serratia rubidae, Citrobacter spp, Enterobacter spp and Salmonella typhi. Disk diffusion antibiotic susceptibility testing was carried out, isolates were screened for ESBL production and confirmed using standard laboratory procedures. ESBLs resistance genes were identified by Polymerase Chain Reaction (PCR). Results: All isolates demonstrated multiple antibiotic resistance. Resistance to ampicillin, amoxicillin with clavulanate and erythromycin was 100%, whereas resistance to Imipenem was very low (5.0%). : Overall prevalence of ESBL producers was 41.4% with Klebsiellaspp as the highest ESBL producing Enterobacteriacaea. ESBL producers were more prevalent among the hospital pathogens than community pathogens, 58% vs 29.5% (p=0.003). ESBL genes were detected in all ESBL producers with the blaCTX-Mgene predominating (47.0%) followed by blaTEM(30.9%) and blaSHVgene was the least, 22.1%. The blaCTX-Mgene was also the most prevalent in the healthcare pathogens (62%) but it accounted for only 25% in those of community origin. Conclusion: A high prevalence of ESBL producing gram negative organisms occurs both in healthcare and in the community in our environment with the CTX-M variant predominating. Efforts to control spread of these pathogens should be addressed.


2017 ◽  
Vol 10 (1) ◽  
pp. 8-12
Author(s):  
Shikha Paul ◽  
Sanya Tahmina Jhora ◽  
Prashanta Prasun Dey ◽  
Bilkis Ara Begum

Detection of Extended spectrum beta lactamase (ESBL) enzyme producing bacteria in hospital settings is vital as ESBL genes are transmissible. This study was carried out to determine the distribution of ESBL producing gram negative isolates at a tertiary care hospital in Dhaka city which deals with the patients hailing from relatively low socioeconomic status.Onehundred and twenty four gram negative bacteria isolated from different clinical specimens from outpatient and inpatient departments of Sir Salimullah Medical College and Mitford Hospital (SSMC & MH) were tested for ESBL by E test ESBL method in the department of microbiology of Sir Salimullah medical college (SSMC) from March 2013 to August 2013.Out of 124 gram negative bacteria 69 (55.65%) were positive for ESBL. Among the ESBL producers, Esch.coli was the highest (46.38%) which was followed by Serratia spp (11.59%), Enterobacter spp (10.14%), Proteus spp, (8.70%), Acinetobacter spp.(7.24%) and Klebsiella spp.(5.79%). Out of 32 Esch.coli isolated from outpatient department, 10 (31.25%) were positive for ESBL. On the other hand out of 27 Esch. coli isolated from inpatient department, 22 (81.48%) were positive for ESBL. The difference was statistically significant (p<0.001).So the present study reveals that the distribution of ESBL producers is more among the hospitalized patients than the patients of the community.Bangladesh J Med Microbiol 2016; 10 (1): 8-12


Author(s):  
Mutasim E. Ibrahim ◽  
Mohammed Abbas ◽  
Abdullah M. Al-Shahrai ◽  
Bahaeldin K. Elamin

Background. Emergence of pathogenic bacteria carrying β-lactamase-resistant determinants has become a major health problem in the hospital setting. The study aimed to determine antibiotic-resistant patterns and frequency of extended-spectrum β-lactamase- (ESBL-) producing Gram-negative bacteria (GNB) and AmpC β-lactamase-producing GNB. Methodology. A prospective cross-sectional study was conducted during a period from September 2017 to August 2018 at King Abdullah Hospital, Bisha Province, Saudi Arabia. GNB (n = 311) were recovered from patients’ clinical specimens including sputum, urine, wound pus, blood, tracheal aspirates and high vaginal swabs, umbilical discharge, eye discharge, and cerebrospinal fluids. Isolates were identified by the Phoenix identification system. Antimicrobial susceptibility was tested by the Kirby–Bauer disk procedure. Phenotypic characterization of ESBLs and AmpC β-lactamases was performed utilizing the double-disk synergy test and inhibitor-based method, respectively. Associations with outcome measures were determined by simple descriptive statistics and a chi-square test. Results. Out of 311 GNB isolates, the frequency of ESBL and AmpC β-lactamase producers was 84 (27%) and 101 (32.5%), respectively. Klebsiella pneumoniae and Escherichia coli were common ESBL producers. AmpC β-lactamases predominate among Acinetobacter spp. and Pseudomonas aeruginosa. Coproduction of ESBLs and AmpC β-lactamases was found in 36 (11.6%) isolates, with very close relative frequencies among K. pneumoniae, Acinetobacter spp., and P. aeruginosa. β-Lactamase producers were predominantly found in the surgical department (56.5%) and ICUs (44.2%). ESBL producers revealed high resistance for cefuroxime (96.4%), cefotaxime (92.9%), and trimethoprim/sulfamethoxazole (90.5%). The resistance rates were significantly higher among ESBL producers than nonproducers for cephalosporins (p<0.001), amoxicillin/clavulanate (p<0.001), piperacillin/tazobactam (p=0.010), nitrofurantoin (p=0.027), aztreonam (p<0.001), ciprofloxacin (p=0.002), and trimethoprim/sulfamethoxazole (p<0.001). Significantly higher (p<0.05) resistance rates were observed among AmpC β-lactamase producers than nonproducers for all tested antibiotics. Conclusions. This finding showed a high prevalence of ESBL- and AmpC β-lactamase-producing GNB in our hospital. Quality control practice and routine detection of β-lactamase producers before deciding on antibiotic therapy are advocated.


Sign in / Sign up

Export Citation Format

Share Document