scholarly journals A 3×4 drop-plating protocol for estimation of Antimicrobial-resistant bacteria, taking Extended-spectrum beta-lactamases producing Escherichia coli as an example.

2021 ◽  
Author(s):  
Yang Zhong
Keyword(s):  
Escherichia Coli ◽  
Risk Assessment ◽  
Resistant Bacteria ◽  
Beta Lactamases ◽  
E Coli ◽  
Colony Counting ◽  
Plating Method ◽  
Extended Spectrum Beta Lactamases ◽  
Direct Spread ◽  
Synergy Test

Abstract The estimation of antimicrobial-resistant (AMR) bacteria plays an important role in risk assessment and surveillance. To test the concentration of resistant bacteria with colony count is a fast and straightforward way to perform. Here we describe an optimized drop-plating method for colony counting of resistant bacteria. We took the ESBL-producing E. coli in freshwater samples as an example. The optimized methods can successfully quantify ESBL-producing E. coli of water samples in a concentration range of 104 CFU/L to 106 CFU/L. We have shown that this drop-plating method is comparable to the direct spreading method by testing with both methods on a series of simulated samples, which were constructed using raw surface water spiked with different concentrations of ESBL-producing E. coli. The ESBL-producing phenotype has been further confirmed with the double-disc synergy test. Compared to direct spread methods, our methods can save consumables and operate with smaller sample sizes. Therefore, this method could be more sustainable in AMR surveillance and risk assessment.

scholarly journals Deteksi Fenotipik Escherichia coli Penghasil Extended Spectrum Beta-lactamases (ESBLs) pada Sampel Makanan di Krian Sidoarjo

Life Science ◽  
2019 ◽  
Vol 8 (1) ◽  
pp. 95-105
Author(s):  
Yulianto Ade Prasetya ◽  
Ike Yuyun Winarsih ◽  
Kharisma Aprilia Pratiwi ◽  
Merinsa Chorry Hartono ◽  
Dita Nur Rochimah
Keyword(s):  
Escherichia Coli ◽  
Methylene Blue ◽  
Food Samples ◽  
Beta Lactamases ◽  
E Coli ◽  
Extended Spectrum ◽  
Confirmation Test ◽  
Extended Spectrum Beta Lactamases ◽  
Fried Foods ◽  
Synergy Test

Escherichia coli is a group of Enterobacteriaceae bacteria that often contaminate food so that it can cause diarrhea. These bacteria are very difficult to treat if they are able to produce the Extended-Spectrum Beta-Lactamases (ESBLs) enzyme. The purpose of this study was to identify ESBLs-producing E. coli in food samples in Krian Sidoarjo. Food samples (fried foods, cilok tempura and chili sauce) were collected from ten different places. The sample was then grown on Eosin Methylene Blue (EMB) medium and purified by the 16 streak method, as well as biochemical character tests. The ESBLs phenotypic E. coli method was carried out by screening test and confirmation test using a Double Disk Synergy Test (DDST). Thirty colonies were able to grow on EMB media, but after microscopic identification and biochemistry testing only four samples were E. coli positive and were able to produce ESBLs from the phenotypic test that had been carried out. ESBLs-producing E. coli testing is important not only for nosocomial infections but also for the community so it needs attention to the spread of ESBLs resistance among microorganism species.   Escherichia coli termasuk kelompok bakteri Enterobacteriaceae yang sering mengkontaminasi makanan sehingga dapat menyebabkan diare. Bakteri ini sangat sulit diobati apabila mampu memproduksi enzim Extended Spectrum Beta-Lactamases (ESBLs). Tujuan penelitian ini adalah untuk mengindetifikasi E. coli penghasil ESBLs pada sampel makanan di Krian Sidoarjo. Sampel makanan (gorengan, cilok, tempura, dan saus sambal) dikumpulkan dari sepuluh tempat berbeda. Sampel kemudian ditumbuhkan pada medium Eosin Metilen Blue (EMB) dan dimurnikan dengan metode streak 16, serta dilakukan karakteristik uji biokimia. Metode fenotipik E. coli penghasil ESBLs dilakukan dengan uji skrining dan uji konfirmasi menggunakan double disk synergy test (DDST). Sebanyak tiga puluh koloni mampu tumbuh pada media EMB, namun setelah diidentifikasi mikroskopis dan uji biokiomia hanya empat sampel positif E. coli dan mampu menghasilkan ESBLs dari uji fenotipik yang telah dilakukan. Pengujian E. coli penghasil ESBLs penting dilakukan bukan hanya pada infeksi nosokomial, tetapi juga pada komunitas sehingga perlu mendapat perhatian terhadap penyebaran resistensi ESBLs diantara spesies mikroorganisme.

scholarly journals ESBL-Producing Escherichia coli Carrying CTX-M Genes Circulating among Livestock, Dogs, and Wild Mammals in Small-Scale Farms of Central Chile

Antibiotics ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 510
Author(s):  
Julio A. Benavides ◽  
Marília Salgado-Caxito ◽  
Andrés Opazo-Capurro ◽  
Paulina González Muñoz ◽  
Ana Piñeiro ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Resistant Bacteria ◽  
Small Scale ◽  
Middle Income ◽  
Central Chile ◽  
Wild Mammals ◽  
Beta Lactamases ◽  
E Coli ◽  
Fecal Carriage ◽  
Extended Spectrum Beta Lactamases

Antibiotic-resistant bacteria of critical importance for global health such as extended-spectrum beta-lactamases-producing (ESBL)-Escherichia coli have been detected in livestock, dogs, and wildlife worldwide. However, the dynamics of ESBL-E. coli between these animals remains poorly understood, particularly in small-scale farms of low and middle-income countries where contact between species can be frequent. We compared the prevalence of fecal carriage of ESBL-E. coli among 332 livestock (207 cows, 15 pigs, 60 horses, 40 sheep, 6 goats, 4 chickens), 82 dogs, and wildlife including 131 European rabbits, 30 rodents, and 12 Andean foxes sharing territory in peri-urban localities of central Chile. The prevalence was lower in livestock (3.0%) and wildlife (0.5%) compared to dogs (24%). Among 47 ESBL-E. coli isolates recovered, CTX-M-group 1 was the main ESBL genotype identified, followed by CTX-M-groups 2, 9, 8, and 25. ERIC-PCR showed no cluster of E. coli clones by either host species nor locality. To our knowledge, this is the first report of ESBL-E. coli among sheep, cattle, dogs, and rodents of Chile, confirming their fecal carriage among domestic and wild animals in small-scale farms. The high prevalence of ESBL-E. coli in dogs encourages further investigation on their role as potential reservoirs of this bacteria in agricultural settings.

scholarly journals Prevalence Of Extended Spectrum Beta-Lactamases (ESBLs)-Producing Escherichia Coli Isolated From UTI Patients Attending some Selected Hospitals In Minna, Nigeria

2021 ◽  
Vol 37 (2) ◽  
pp. 56-73
Author(s):  
F Iseghohi ◽  
J.C Igwe ◽  
M Galadima ◽  
A.F Kuta ◽  
A.M Abdullahi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Multidrug Resistance ◽  
Urine Samples ◽  
Antibiotics Resistance ◽  
Healthy Individuals ◽  
Beta Lactamases ◽  
E Coli ◽  
Extended Spectrum ◽  
Extended Spectrum Beta Lactamases ◽  
Tract Infections

Globally, urinary tract infections are one of the most common infections in need of urgent clinical attention. The prevalence of extended spectrum beta-lactamases (ESBL)- producing Escherichia coli isolated from urine samples of some UTI patients and s of apparently healthy individuals in Minna, Nigeria, is investigated. Standard microbiological techniques were used to conduct this study. A total of 170 catch midstream urine samples submitted to the Medical Microbiology Laboratories of 4 different hospitals (and samples from healthy individuals) were randomly collected for 5 months and examined for microbial growths. Female patients (65.9%) submitted more urine samples for UTI test than their male counterpart (34.1%). The age ranges of 21 -30 (26.5%) and 31 - 40 (25.3%) had the highest percentages of infection rate while those within the ages 1- 10 (3.5%) and ≥ 71 (2.3%) were the least infected. This study observed a prevalence of 23.5% of E. coli in Minna metropolis and a significant number (30%) of healthy individuals (HI) was observed to harbor the E. coli in their urine. The isolates were highly susceptible to Gentamicin (65%), Ofloxacin (65%), Tetracycline (62.5%), Cotrimoxazole (62.5%), and Streptomycin (57.5%). Mildly susceptible to Pefloxacin (37.5%), Chloramphenicol (37.5%), and Ciprofloxacin (35%). There were significant resistance to most of the beta-lactames tested [Cefuroxime (80%), Amoxicillin (42.5%), Augmentin (40), Cefotaxime (20%) and Ceftaxidime (7.5%)]. Two of the isolates were resistant to all the 13 antibiotics tested; 70% (28) of the isolates had multiple antibiotics resistance index (MARI) ≥0.3. Multidrug resistance was expressed in 37.5% of the isolates tested. The study showed a vast resistant pool in the environment. Only 25% of the E. coli isolated from the urine samples produced beta-lactamases phenotypically, most of which expressed resistance to more than 5 of the antibiotics tested and had MARI of ≥ 0.5. Further evaluation showed that 25% (10/40) of the E. coli isolated from the UTI patients in Minna, Nigeria, were ESBL- producers and could harbor one or two of the genes. TEM gene was expressed in 70% (7) of the isolates that produced ESBL phenotypically, 60% 6) harbored CTXM gene, 20% (2) had the OXA gene while none of the bacteria harbored the SHV gene. The study established a 5.9% ESBL prevalence among the E. coli isolated from UTI in the environment studied. This study established that E. coli is one of the prevalent bacteri urea majorly isolated from UTI patients in Minna. The prevalent E. coli are multidrug resistant and could harbor more than one ESBL gene . keywords: Escherichia coli, Minna, UTI, ESBL, Multidrug resistance

scholarly journals Improvement of Antibiotics Susceptibility of Escherichia coli in a Tertiary Hospital in Japan

2019 ◽  
pp. 1-5
Author(s):  
Yuji Watanabe ◽  
Masafumi Seki
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Tertiary Hospital ◽  
Clinical Settings ◽  
Infection Control Team ◽  
Beta Lactamases ◽  
E Coli ◽  
Control Team ◽  
Extended Spectrum Beta Lactamases ◽  
Antibiotics Susceptibility

Antimicrobial stewardship team (AST) and Infection Control Team (ICT) have recently been linked Infectious diseases (ID) physicians, and implemented in clinical settings in Japan. The microbiological effects of an AST and ICT, in addition to Diagnostic stewardship team (DST) supported by ID physicians in our tertiary hospital were shown in significant reduction of antibiotic resistance of Escherichia coli (E coli) including extended spectrum beta-lactamases (ESBL)-producing E coli.

scholarly journals Characterization of ESBL-Producing Enterobacteria from Fruit Bats in an Unprotected Area of Makokou, Gabon

2020 ◽  
Vol 8 (1) ◽  
pp. 138 ◽  
Author(s):  
Pierre Philippe Mbehang Nguema ◽  
Richard Onanga ◽  
Guy Roger Ndong Atome ◽  
Jean Constant Obague Mbeang ◽  
Arsène Mabika Mabika ◽  
...  
Keyword(s):  
Resistant Bacteria ◽  
Fruit Bats ◽  
Antibiotic Resistant ◽  
Disk Diffusion Test ◽  
Beta Lactamases ◽  
E Coli ◽  
Terrestrial Mammals ◽  
Extended Spectrum Beta Lactamases ◽  
First Time

In Gabon, terrestrial mammals of protected areas have been identified as a possible source of antibiotic-resistant bacteria. Some studies on antibiotic resistance in bats have already been carried out. The main goal of our study was to detect extended-spectrum beta-lactamases (ESBLs) that are produced by enterobacteria from bats in the Makokou region in Gabon. Sixty-eight fecal samples were obtained from 68 bats caught in the forests located 1 km from the little town of Makokou. After culture and isolation, 66 Gram-negative bacterial colonies were obtained. The double-disk diffusion test confirmed the presence of ESBLs in six (20.69%) Escherichia coli isolates, four (13.79%) Klebsiella pneumoniae isolates, and one (3.45%) Enterobacter cloacae isolate. The analysis based on the nucleotide sequences of the ESBL resistance genes showed that all cefotaximase-Munichs (CTX-Ms) were CTX-M-15 and that all sulfhydryl variables (SHVs) were SHV-11: 41.67% CTX-M-15-producing E. coli, 16.67% CTX-M-15+SHV-11-producing E. coli, 8.33% CTX-M-15-producing K. pneumoniae, 25% CTX-M-15+SHV-11-producing K. pneumoniae, and 8.33% CTX-M-15-produced E. cloacae. This study shows for the first time the presence of multiresistant ESBL-producing enterobacteria in fruit bats in Makokou.

scholarly journals Dogs of Nomadic Pastoralists in Northern Kenya Are Reservoirs of Plasmid-Mediated Cephalosporin- and Quinolone-Resistant Escherichia coli, Including Pandemic Clone B2-O25-ST131

2012 ◽  
Vol 56 (7) ◽  
pp. 4013-4017 ◽  
Author(s):  
Katerina Albrechtova ◽  
Monika Dolejska ◽  
Alois Cizek ◽  
Dagmar Tausova ◽  
Jiri Klimes ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Content Type ◽  
Beta Lactamases ◽  
Northern Kenya ◽  
E Coli ◽  
Nomadic Pastoralists ◽  
Extended Spectrum ◽  
Extended Spectrum Beta Lactamases ◽  
Pfge Profiles

ABSTRACTResistance inEscherichia coliisolates colonizing gastrointestinal tracts of dogs, cats, and their owners in Northern Kenya was investigated with an emphasis on extended-spectrum beta-lactamases (ESBLs). Totals of 47 (22%,n= 216), 2 (4%,n= 50), and 4 (17%,n= 23) CTX-M-15-producingE. coliisolates were obtained from dogs, cats, and humans, respectively. CTX-M-15-producingE. coliisolates with identical PFGE profiles were detected in animals and humans living in the same area.

scholarly journals Silver Resistance Genes Are Overrepresented among Escherichia coli Isolates with CTX-M Production

2014 ◽  
Vol 80 (22) ◽  
pp. 6863-6869 ◽  
Author(s):  
Susanne Sütterlin ◽  
Petra Edquist ◽  
Linus Sandegren ◽  
Marlen Adler ◽  
Thomas Tängdén ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Resistance Genes ◽  
Selective Pressure ◽  
Human Origin ◽  
Content Type ◽  
Beta Lactamases ◽  
E Coli ◽  
Genes Encoding ◽  
Extended Spectrum Beta Lactamases ◽  
M 14

ABSTRACTMembers of theEnterobacteriaceaewith extended-spectrum beta-lactamases (ESBLs) of the CTX-M type have disseminated rapidly in recent years and have become a threat to public health. In parallel with the CTX-M type expansion, the consumption and widespread use of silver-containing products has increased. To determine the carriage rates of silver resistance genes in differentEscherichia colipopulations, the presence of three silver resistance genes (silE,silP, andsilS) and genes encoding CTX-M-, TEM-, and SHV-type enzymes were explored inE. coliisolates of human (n= 105) and avian (n= 111) origin. The antibiotic profiles were also determined. Isolates harboring CTX-M genes were further characterized, and phenotypic silver resistance was examined. ThesilEgene was present in 13 of the isolates. All of them were of human origin. Eleven of these isolates harbored ESBLs of the CTX-M type (P= 0.007), and eight of them were typed as CTX-M-15 and three as CTX-M-14. None of thesilE-positive isolates was related to the O25b-ST131 clone, but 10 out of 13 belonged to the ST10 or ST58 complexes. Phenotypic silver resistance (silver nitrate MIC > 512 mg/liter) was observed after silver exposure in 12 of them, and a concomitant reduced susceptibility to piperacillin-tazobactam developed in three. In conclusion, 12% of the humanE. coliisolates but none of the avian isolates harbored silver resistance genes. This indicates another route for or level of silver exposure for humans than that caused by common environmental contamination. SincesilE-positive isolates were significantly more often found in CTX-M-positive isolates, it is possible that silver may exert a selective pressure on CTX-M-producingE. coliisolates.

scholarly journals Extended-spectrum beta-lactamases producing Escherichia coli and Klebsiella pneumoniaei: A Multi-centric Study Across Karnataka

2014 ◽  
Vol 6 (01) ◽  
pp. 007-013 ◽  
Author(s):  
Sridhar PN Rao ◽  
Prasad Subba Rama ◽  
Vishwanath Gurushanthappa ◽  
Radhakrishna Manipura ◽  
Krishna Srinivasan
Keyword(s):  
Escherichia Coli ◽  
Clinical Isolates ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Esbl Production ◽  
Beta Lactamases ◽  
E Coli ◽  
Karnataka State ◽  
Extended Spectrum Beta Lactamases ◽  
Esbl Producers

ABSTRACT Background: There are sporadic reports on detection of extended-spectrum beta-lactamases (ESBL) producers from Karnataka; hence, this is a first multicentric study across Karnataka state to determine the prevalence of ESBL production among clinical isolates of Escherichia coli and Klebsiella pneumoniaei. Aims and objectives: To determine the prevalence of ESBL producing clinical isolates of E. coli and K. pneumoniae from five geographically distributed centers across Karnataka, to study the susceptibility of ESBL producing isolates to other beta-lactam and beta-lactam-beta-lactamase inhibitors and to demonstrate transferability of plasmids coding for ESBL phenotype. Materials and Methods: Two hundred isolates of E. coli and K. pneumoniae each were collected from each of the five centers (Bellary, Dharwad, Davangere, Kolar and Mangalore). They were screened for resistance to screening agents (ceftazidime, cefotaxime, ceftriaxone, aztreonam) and positive isolates were confirmed for ESBL production by test described by Clinical and Laboratory Standards Institute . Co-production of ESBL and AmpC beta-lactamase was identified by using amino-phenylboronic acid disk method. Susceptibility of ESBL producers to beta-lactam antibiotics and beta-lactamase inhibitors was performed. Transferability of plasmids was performed by conjugation experiment. Results: Overall prevalence of ESBL production among E. coli and K. pneumoniae across five centers of the state was 57.5%. ESBL production was found to be 61.4% among E. coli and 46.2% among K. pneumoniae. ESBL production was significantly more among E. coli than K. pneumoniae. Significant variations in distribution of ESBL across the state was observed among E. coli isolates, but not among K. pneumoniae isolates. All ESBL producers demonstrated minimum inhibitory concentration levels ≥2 μg/ml towards cefotaxime, ceftazidime and ceftriaxone. Conclusion: Overall prevalence of ESBL production among clinical isolates of E. coli and K. pneumoniae across Karnataka state was high. The prevalence of ESBL production was significantly higher with E. coli than K. pneumoniae isolates. Higher rates of resistance to ceftriaxone and cefotaxime than to ceftazidime suggests the possibility of presence of CTX-M type ESBLs. Of all the beta-lactam/beta-lactamase inhibitor combinations tested, cefepime-tazobactam demonstrated highest in-vitro activity against ESBL producers. There was no statistical difference in the transferability of plasmids among E. coli and K. pneumoniae.

scholarly journals Genetic Investigation of Beta-Lactam Associated Antibiotic Resistance Among Escherichia Coli Strains Isolated from Water Sources

2017 ◽  
Vol 11 (1) ◽  
pp. 203-210 ◽  
Author(s):  
Reza Ranjbar ◽  
Mehrdad Sami
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Surface Water ◽  
Water Resources ◽  
Water Sources ◽  
Resistant Bacteria ◽  
Beta Lactam ◽  
E Coli ◽  
Surface Water Resources ◽  
Extended Spectrum Beta Lactamases

Background: Antimicrobial resistance is an important factor threatening human health. It is widely accepted that antibiotic resistant bacteria such as Escherichia coli (E. coli) released from humans and animals into the water sources, can introduce their resistance genes into the natural bacterial community. Objective: The aim of this study was to investigate the prevalence of blaTEM, blaCTX, blaSHV, blaOXA and blaVEB associated-antibiotic resistance among E. coli bacteria isolated from different water resources in Iran. Methods: The study contained all E. coli strains segregated from different surface water sources. The Kirby-Bauer method and combined discs method was determined in this study for testing antimicrobial susceptibility and strains that produced Extended-Spectrum Beta Lactamases (ESBL), respectively. DNA extraction kit was applied for genomic and plasmid DNA derivation. Finally the frequency of resistant genes including blaTEM, blaCTX, blaSHV, blaOXA and blaVEB in ESBL producing isolates were studied by PCR. Results: One hundred E. coli strains were isolated and entered in the study. The highest antibiotic resistance was observed on clindamycin (96%). Moreover, 38.5% isolates were ESBL producers. The frequency of different ESBLs genes were 37%, 27%, 27%, and 25% for blaTEM, blaCTX, blaSHV, and blaOXA, respectively. The blaVEB wasn’t found in any isolates. Conclusion: The study revealed a high prevalence of CTX-M, TEM, SHV and OXA genes among E. coli strains in surface water resources. In conclusion, these results raised a concern regarding the presence and distribution of these threatening factors in surface water sources and its subsequent outcomes.

scholarly journals PREVALENCE OF EXTENDED SPECTRUM BETA-LACTAMASES (ESBLS) PRODUCING ESCHERICHIA COLI AND KLEBSIELLA PNEUMONIAE AMONG HOSPITALIZED PATIENTS FROM NIGERIA

2021 ◽  
Vol 5 (2) ◽  
pp. 584-595
Author(s):  
Olivia Sochi Egbule ◽  
Bernard O. Ejechi
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
General Hospital ◽  
Hospitalized Patients ◽  
Diffusion Method ◽  
Alkaline Lysis ◽  
Beta Lactamases ◽  
Resistance Patterns ◽  
Extended Spectrum Beta Lactamases ◽  
Synergy Test

The aim of this study was to determine the resistance patterns and ESBLs production among clinical isolates of Escherichia coli and Klebsiella pneumoniae in two government hospitals of Delta State, Nigeria. Urine, blood and wound samples were aseptically collected from hospitalized patients, bacteriologically processed and isolates identified using standard protocols. Antimicrobial susceptibility testing was determined by disc diffusion method. The plasmid DNA of Multidrug resistance (MDR) isolates were extracted by alkaline lysis method. Phenotypic ESBL production of the MDR isolates was done by Double Disc Synergy Test (DDST) while PCR was used to detect blaCTX-M, blaSHV and blaTEM among isolates. A total of 217 isolates were obtained, of which 161(74.2%) and 56(25.8%) were Escherichia coli and Klebsiella pneumoniae respectively. The antimicrobial resistance varied from one location to another. All isolates obtained from blood of general hospital Warri (GHW) were 100% resistant to amoxicillin clavulanic acid and the cephalosporins (ceftazidime, cefotaxime, and cefuroxime). Isolates from General hospital Agbor (GHA) showed high resistance of 75.0% to cefotaxime, 93.8% to each of ceftazidime and cefuroxime. Overall low resistance to nitrofurantoin was observed in E. coli isolates obtained from urine of GHW (27.5%) and GHA (20.8%). Out of 217 isolates, 75.1% (163/217) were MDR, of which 36.8% and 39.3% produced ESBL by DDST and PCR respectively. The most common ESBL gene was blaCTX-M expressed by 28(17.2%) of the isolates. The high prevalence of MDR and ESBL underscores the need for a continuous local monitoring of antibiotic resistance.    

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