scholarly journals Evaluation of different phenotypic diffusion methods in the identification of extended spectrum beta lactamase producing uropathogenic

2021 ◽  
Vol 8 (3) ◽  
pp. 243-248
Author(s):  
H Anandkumar ◽  
Amaresh Nigudgi ◽  
Vinay Hajare ◽  
Sunil Biradar

: The study was aimed to identify the occurrence of extended spectrum of Beta lactamases (ESBLs), to compare different phenotypic methods used for the confirmation and to evaluate the antibiotic resistance pattern in ESBL producing uropathogenic : The strains were isolated from urine and the isolates resistance to at least one of the three representative cephalosporins (cefotaxime, cefpodoxime and ceftazidime) was tested for ESBL production by Double disc synergy test (DDST), Inhibitory potentiated disc diffusion (IPDD) test and quantitative E-strip method.: Of 120strains isolated, 62(51.6%) were resistant to at least one of the three cephalosporins and 28 (45.1%) were positive for ESBL by IPDD and E-strip test. However, 9 (14.5%) strains were positive by DDST method. Among third generation cephalosporins, cefpodoxime was (45.8%) better screening indicator followed by ceftazidime (40.0%) and cefotaxime (37.5%). Most of the ESBL producers (97.3%) were resistant to three or more drugs, compared to (51.2%) non-ESBL producers. : The acceptable method for detection of ESBL producing were IPDD and E-strip tests compared to DDST with better sensitivity (100%), specificity (95.8%) and positive predictive value (96.5%). ESBL producers showed significantly (p<0.05) higher resistance to tobramycin, amoxyclav and amikacin compared to non ESBL producers.

2021 ◽  
Vol 37 (2) ◽  
pp. 113-125
Author(s):  
M.K Salihu ◽  
A Yarima ◽  
H.I Atta

1983. These enzymes possess the ability to inactivate susceptible β-lactam antibiotics i.e. penicillins, first, second and third generation cephalosporins and aztreonam, but not cephamycins and carbapenems . Their mode of action is by hydrolyzing the β-lactam ring. Even before the first β-lactam antibiotic (penicillin) was developed, resistance to β-lactam antibiotics was observed . ESBL genes are plasmids- and transposons- mediated, as such, can be spread easily to other species of bacteria. Resistance of ESBL- producing bacteria to the β-lactam antibiotics is a continuing cause of public health problems , it is increasingly being observed in community and nosocomial acquired infections. Detection and identification of these ESBLs in the laboratory is of prime importance for the selection of appropriate antibiotics to be used in the treatment of infections caused by ESBL- producing bacteria. The aim of this review is to explain in detail , several phenotypic methods used in the detection and confirmation of extended spectrum β lactamases. Keywords: Antibiotic resistance, ESBL, bacteria, phenotypic method


2020 ◽  
Vol 14 (8) ◽  
pp. 2746-2757
Author(s):  
Souleymane Soré ◽  
Yacouba Sawadogo ◽  
Juste Isidore Bonkoungou ◽  
Sephora P. Kaboré ◽  
Saidou Béogo ◽  
...  

Extended spectrum beta-lactamase producing Enterobacteriaceae (ESBL-PE) represent a threat for failure of empirical antibiotic therapy and are associated with high mortality, morbidity and expenses. The aims of this study was to determine the prevalence of ESBL-PE and multidrug resistant enterobacteria (MDR), enterobacteria profil, investigate the associated resistance in wastewater and salads. After wastewater and salad sampling, enterobacteria was isoled on (EMB + 4μg / L cefotaxim). The stains of Enterobacteriaceae were identified by using biochemical methods and confirmed as ESBL by double-disc synergy test (amoxicillin/clavulanic acid with cefotaxime 30 μg, ceftazidime 30 μg and ceftriaxone 30 μg). Finally, the associated resistance was investigated by testing the susceptibility of the strains by the disc diffusion method. Global prevalence of ESBL-PE was 53.92% (95% CI: 48,2-59,5) (153/293), 61.11% from wastewater and 42.47% from salads. Major ESBL-E was Escherichia coli (73.44%), followed by Klebsiella pneumoniae (21.88%). Resistance to the aminoglycoside , fluroquinolonones and sulfonamides classes were dominant, observed in 53,83%, 93,86% and 98,95% of the isolates, respectively. The frequence of MDR was hight to channel1 (32,40%) and channel2 (26,26%). This study reports very worrying results. There is an urgent need to develop measures to monitor the spread of these multidrug-resistant strains.Keywords: Wastewater, ESBL-PE, Salads, Ouagadougou.


2021 ◽  
Vol 57 (4) ◽  
pp. 283
Author(s):  
Ardhiya Puspita ◽  
Radita Yuniar Arizandy ◽  
Eddy Bagus Wasito ◽  
Kuntaman Kuntaman

Highlight :Bacteriologically for colonization of  ESBL producing Enterobacteriaceae in cockroaches (Periplaneta americana) were analyzed.The prevalence of ESBL producing bacteria among cockroaches in hospitals is bigger than in households.Abstract: Cockroach (Periplaneta americana) is one of the vectors in the environment that can transmit disease. Cockroaches can act as potential mechanical vectors of antibiotic resistant bacteria. Enterobacteriaceae is a gram-negative bacteria that has natural habitats in the digestive tract of humans and animals. Enterobacteriaceae that produce Extended Spectrum β-lactamases (ESBLs) have emerged as major pathogens in hospitals. The study analyzed the prevalence of ESBL producing bacteria in cockroaches that lived in hospitals and residential homes. In this study, a total of 200 cockroaches consisting of 100 cockroaches from the hospital environment and 100 cockroaches from the residential environment were analyzed bacteriologically for colonization of  ESBL producing Enterobacteriaceae. The specimen of the alimentary tract was taken and sub-cultured in MacConkey agar supplemented with cefotaxime 2 ug/ml. Growth colonies were suggested as an ESBL-producing bacteria, then were confirmed as ESBL producers by the Double Disk Synergy Test (DDST). The ESBL gene was detected by Polymerase Chain Reaction (PCR). Among 100 household cockroach samples, 14 (14%) were identified as ESBL producers, while 100 hospital cockroaches were 26 (26%) positive ESBL. The ESBL gene, in hospital cockroach were identified of CTXM 19 (19%), SHV 7 (7%), and not any TEM gene, while among household cockroaches were identified CTXM 2 (2%), SHV 11 (11%), and also not detected TEM ESBL gene. Among ESBL genes, only the CTXM gene was significantly different between household and hospital cockroaches.


Author(s):  
F. Z. Uyanga ◽  
E. O. Ekundayo ◽  
E. O. Nwankwo ◽  
A. I. Inimfon

Background/Purpose: The aim of this study was to evaluate the effectiveness of CHROMagar ESBL in comparison with Double Disc Synergy Test (DDST) for the detection of ESBL producing uropathogens. Methods: Six hundred and sixty urine samples were collected from pregnant women attending antenatal at General hospital Ikot Ekpene, Eket and Oron. Two hundred and fifty eight isolates were obtained while two hundred and thirty one isolates were ESBL producers. Microbact 24E(Oxoid, UK) was used in the identification of bacterial isolates, antibiotic susceptibility test was done using Kirby-Bauer disk diffusion method following CLSI guidelines using commercially available disc (Oxoid Ltd). Double disk synergy test was carried out on the isolates and inoculation was done using CHROMagar ESBL (France). Results: The prevalence of ESBL 35% was recorded. The sensitivity and specificity of DDST was 88% and 89%, respectively. CHROMagar showed an increase in sensitivity and specificity at 48 h with 98% and 99.0%, respectively. 80% of the ESBL producing isolates were multi drugs resistant. The predominant bacterial pathogens were Enterobacter cloacae (23%), Proteus mirabilis (14%) and Acinetobacter baumanii (13.4%).The ESBL producing isolates showed maximum resistance against Ceftazidime (90%), Cefotaxime (91%), Azetronam (95%), Amikacin (68.2%) followed by ofloxacin (70%) while maximum sensitivity was seen for imipenem (90%) and Augumentin (80%). The study demonstrated that CHROMagar was superior and more sensitive than DDST. Conclusion: CHROMagar ESBL seems to be the most reliable method among phenotypic methods for detection of ESBL in the absence of PCR.


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.


2015 ◽  
Vol 17 (3) ◽  
Author(s):  
Debora C. Kajeguka ◽  
Petro P. Nambunga ◽  
Frank Kabissi ◽  
Benjamin Kamugisha ◽  
Nancy Kassam ◽  
...  

Background: Production of Extended Spectrum Beta-Lactamase (ESBL) by bacteria is a chronic problem in a health care set up. In order to have adequate information for treatment of bacterial infections especially ESBL producing isolates, it is crucial to understand the trends in the antibiotic-resistance pattern, occurrence and their geographical spread.  The objective of this study was to determine the antimicrobial resistance pattern among phenotype ESBL producing isolates in northern Tanzania.Methods: From July 2013 to January 2014, urine, pus and blood samples were collected from patients suspected to have bacterial infections at Kilimanjaro Christian Medical Centre in Moshi, Tanzania. The isolates were identified based on standard laboratory procedures. Antimicrobial susceptibility tests were carried out using various antimicrobial discs as per the recommendations of Clinical Laboratory Standard Institute.Results: A total of 330 specimens were collected. They consisted of 46 urine, 264 pus (from wound) and 20 blood samples. Among isolated bacteria, ESBL producers were 29.7% (98) and non-producers were 70.5% (232).  Escherichia coli and Klebsiella pneumoniae were the most isolated bacteria and dominant ESBL producers.  ESBL production was highly associated with moderate condition at discharge and longer periods of admission. More than 60% of the ESBL producing E. coli were resistant to ceftazidime, cefpodoxime, cefotaxime, amoxycilin, ciprofloxacin, and gentamycin. More than 80% of ESBL producing K. pneumonia and Proteus mirabilis were resistant to ceftazidime and cefotaxime. Fifty four percent of ESBL producing K. pneumonia were resistant to gentamycin.Conclusion: This study shows that ESLB phenotypes among Gram-negative bacteria are common among patients attending a tertiary hospital in northern in Tanzania. The findings suggest that clinical microbiology laboratories should take into account the diagnosis of ESBL producers in order to define the degree of the problem so as to establish a proper treatment protocol.


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.


2006 ◽  
Vol 50 (9) ◽  
pp. 2990-2995 ◽  
Author(s):  
Xiaofei Jiang ◽  
Zhe Zhang ◽  
Min Li ◽  
Danqiu Zhou ◽  
Feiyi Ruan ◽  
...  

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%.


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


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