scholarly journals Detection of CTX-M-type ESBLs from Escherichia coli Clinical Isolates from a Tertiary Hospital, Malaysia

2019 ◽  
Vol 16 (3(Suppl.)) ◽  
pp. 0682 ◽  
Author(s):  
MKK Et al.

The present study aims to detect CTX-M-type ESBL from Escherichia coli clinical isolates and to analyze their antibotic susceptibility patterns. One hundred of E. coli isolates were collected from different clinical samples from a tertiary hospital. ESBL positivity was determined by the disk diffusion method. PCR used for amplification of CTX-M-type ESBL produced by E. coli. Out of 100 E. coli isolates, twenty-four isolates (24%) were ESBL-producers. E. coli isolated from pus was the most frequent clinical specimen that produced ESBL (41.66%) followed by urine (34.21%), respiratory (22.23%), and blood (19.05%).  After PCR amplification of these 24 isolates, 10 (41.66%) isolates were found to possess CTX-M genes. The CTX-M type ESBL producing E. coli against antibiotics belonging to different families showed the highest resistance rates to Ampicillin (100%), Cefotaxime (97%), Cefuroxime (95%), and Ciprofoxacin (86%). Carbapenem groups of antibiotics, Meropenem (89%) and Imipenem (85%) have the highest susceptibility rate among all antibiotics used in this study. The outcome of the antimicrobial susceptibility testing of significant CTX-M- type ESBL producing E. coli could be useful to avoid failure or prolong treatments.

2019 ◽  
Vol 6 (1) ◽  
pp. e000369 ◽  
Author(s):  
Magdalena Nüesch-Inderbinen ◽  
Nadine Käppeli ◽  
Marina Morach ◽  
Corinne Eicher ◽  
Sabrina Corti ◽  
...  

BackgroundEscherichia coli is an important aetiological agent of bovine mastitis worldwide.MethodsIn this study, 82 E. coli from bovine mastitis milk samples from 49 farms were analysed for their genetic diversity using phylogenetic grouping and multilocus sequence typing. The isolates were examined by PCR for a selection of virulence factors (VFs). Antimicrobial susceptibility profiles were assessed using the disk diffusion method.ResultsThe most prevalent phylogroups were group B1 (41.5 per cent of the isolates) and group A (30.5 per cent). A variety of 35 different sequence types (STs) were identified, including ST1125 (11 per cent), ST58 (9.8 per cent), ST10 (8.5 per cent) and ST88 (7.3 per cent). Aggregate VF scores (the number of unique VFs detected for each isolate) ranged from 1 to 3 for 63.4 per cent of the isolates and were at least 4 for 12.2 per cent. For 24.4 per cent of the isolates, the score was 0. The three most frequent VFs were traT, fyuA and iutA. The majority (72 per cent) of the isolates harboured traT. The majority (68.3 per cent) of the isolates were fully susceptible to all antimicrobials tested, with 22 per cent resistant to ampicillin and 14.6 per cent to tetracycline. Resistance rates were low for gentamicin (3.7 per cent), amoxicillin/clavulanic acid (2.4 per cent) and ceftiofur (1.2 per cent), respectively.ConclusionAmong the study’s sample population, E. coli strains were genotypically diverse, even in cows from the same farm, although some STs occurred more frequently than others. Susceptibility to clinically relevant compounds remained high.


2017 ◽  
Vol 66 (2) ◽  
pp. 163-169
Author(s):  
Mona T. Kashef ◽  
Omneya M. Helmy

Aminoglycosides are used in treating a wide range of infections caused by Gram-positive and Gram-negative bacteria; however, aminoglycoside resistance is common and occurs by several mechanisms. Among these mechanisms is bacterial rRNA methylation by the 16S rRNA methyl transferase (16S-RMTase) enzymes; but data about the spread of this mechanism in Egypt are scarce. Cephalosporins are the most commonly used antimicrobial agents in Egypt; therefore, this study was conducted to determine the frequency of 16S-RMTase among third generation cephalosporin-resistant clinical isolates in Egypt. One hundred and twenty three cephalosporin resistant Gram-negative clinical isolates were screened for aminoglycosides resistance by the Kirby Bauer disk diffusion method and tested for possible production of 16S-RMTase. PCR testing and sequencing were used to confirm the presence of 16S-RMTase and the associated antimicrobial resistance determinants, as well as the genetic region surrounding the armA gene. Out of 123 isolates, 66 (53.66%) were resistant to at least one aminoglycoside antibiotic. Only one Escherichia coli isolate (E9ECMO) which was totally resistant to all tested aminoglycosides, was confirmed to have the armA gene in association with blaTEM-1, blaCTX-M-15, blaCTX-M-14 and aac(6)-Ib genes. The armA gene was found to be carried on a large A/C plasmid. Genetic mapping of the armA surrounding region revealed, for the first time, the association of armA with aac(6)-Ib on the same transposon. In conclusion, the isolation frequency of 16S-RMTase was low among the tested aminoglycoside-resistant clinical samples. However, a novel composite transposon has been detected conferring high-level aminoglycosides resistance.


2020 ◽  
Vol 6 (2) ◽  
pp. 147-152
Author(s):  
Ebuka Elijah David ◽  
Muhammad Arfat Yameen ◽  
Ikechuku Okorie Igwenyi ◽  
Arthur Chinedu Okafor ◽  
Uket Nta Obeten ◽  
...  

Aim: This study was aimed to determine the virulent genes and antibiotic resistance patterns among circulating diarrheagenic Escherichia coli (DEC) pathotypes in a tertiary care health center in east of Nigeria. Materials and Methods: Diarrheal stool samples were obtained from 80 children under 5 years and E. coli was isolated and identified using standard biochemical and molecular methods. Multiplex polymerase chain reaction (PCR) was used to detect eight virulent genes of DEC. Disk diffusion method was used to determine the antibiotic susceptibility of DEC. Results: DEC infection was observed in 54 (68%) children among which ial gene for enteroinvasive E. coli (EIEC) (40% [n=22]) was commonly detected followed by eltA/eltB for enterotoxigenic E. coli (ETEC) (30% [n=16]), pCVD for enteroaggregative E. coli (EAEC) (20% [n=11]), and eaeA/bfpA for typical enteropathogenic E. coli (EPEC) (10% [n=5]). The DEC isolates phenotypically exhibited resistance for ampicillin (AMP) (44 [81%]), followed by ciprofloxacin (CIP)/ levofloxacin (LEV) (28 [52%]), cefoxitin (FOX) (11 [20%]), and amoxicillin-clavulanic acid (AMC) (6 [11%]). About 60% isolates of stable toxins-ETEC were resistant to AMC, CIP, and LEV while all the labile toxin-ETEC exhibited resistance to AMP. About 60% (n=6) resistance were seen in EAEC against ampicillin, AMC, FOX, CIP, and LEV. In EIEC, all the isolates (n=22) were resistant to AMP while 50% (n=11) were resistant to both CIP and LEV. All EPEC (n=5) were resistant to AMP, FOX, CIP, and LEV. Conclusion: High frequency of virulent ial and eltA/eltB genes for EIEC and ETEC, respectively, suggests that they are the primary etiological agents of diarrhea in children among DEC pathotypes. Resistance of DEC to more than two classes of antibiotics indicate possible emergence of multidrug resistance.


2021 ◽  
Vol 11 (1) ◽  
pp. 84-92
Author(s):  
René Dembélé ◽  
Issiaka Soulama ◽  
Wendpoulomdé Aimé Désiré Kaboré ◽  
Ali Konaté ◽  
Assèta Kagambèga ◽  
...  

Background and objective: In recent years, carbapenemase-producing Enterobacterales (CPE) resistance to antibiotics has dramatically increased leading to limitations of their treatment options. In the present study, we investigated the occurrence of carbapenemase-producing Escherichia coli and Salmonella in rural Burkina Faso.Materials and methods: Salmonella isolates were serotyped according to the Kauffman White scheme. Diarrheagenic Escherichia coli (DEC) strains was identified using 16-plex Polymerase Chain Reaction (PCR), whereas antibiotic susceptibility was realized using the disk diffusion method. Furthermore, multiplex PCR assays were carried out using oligonucleotides to detect the presence of genes of the blaKPC, blaVIM, blaIMP, blaTEM, blaSHV, blaOXA and blaCTX-M types in all E. coli and Salmonella strains.Results: The study highlighted high resistance rates of the identified bacteria to common antibiotics. Likewise, two strains of E. coli were imipenem resistant with carbapenemase-encoding genes. The genes detected were Klebsiella pneumoniae carbapenemase (KPC), Verona integrin-encoded metallo-β-lactamase (VIM) and Imipenemase (IMP-2) reaching a rate of 40% each in E. coli strains. However, no Salmonella carbapenemases blaKPC, blaVIM or blaIMP were detected.Conclusion: This study showed that for a real-time infection control and prompt application of antimicrobial chemotherapy, characterization of carbapenemase-producing Enterobacterales in patients is crucial.Keywords: Antibiotics, Carbapenemase-Producing Enterobacterales, children, Burkina Faso


2021 ◽  
Vol 9 (2) ◽  
pp. 326
Author(s):  
Frederick Adzitey ◽  
Nurul Huda ◽  
Amir Husni Mohd Shariff

Meat is an important food source that can provide a significant amount of protein for human development. The occurrence of bacteria that are resistant to antimicrobials in meat poses a public health risk. This study evaluated the occurrence and antimicrobial resistance of E. coli (Escherichia coli) isolated from raw meats, ready-to-eat (RTE) meats and their related samples in Ghana. E. coli was isolated using the USA-FDA Bacteriological Analytical Manual and phenotypic antimicrobial susceptibility test was performed by the disk diffusion method. Of the 200 examined meats and their related samples, 38% were positive for E. coli. Notably, E. coli was highest in raw beef (80%) and lowest in RTE pork (0%). The 45 E. coli isolates were resistant ≥ 50% to amoxicillin, trimethoprim and tetracycline. They were susceptible to azithromycin (87.1%), chloramphenicol (81.3%), imipenem (74.8%), gentamicin (72.0%) and ciprofloxacin (69.5%). A relatively high intermediate resistance of 33.0% was observed for ceftriaxone. E. coli from raw meats, RTE meats, hands of meat sellers and working tools showed some differences and similarities in their phenotypic antimicrobial resistance patterns. Half (51.1%) of the E. coli isolates exhibited multidrug resistance. The E. coli isolates showed twenty-two different resistant patterns, with a multiple antibiotic resistance index of 0.0 to 0.7. The resistant pattern amoxicillin (A, n = 6 isolates) and amoxicillin-trimethoprim (A-TM, n = 6 isolates) were the most common. This study documents that raw meats, RTE meats and their related samples in Ghana are potential sources of antimicrobial-resistant E. coli and pose a risk for the transfer of resistant bacteria to the food chain, environment and humans.


2011 ◽  
Vol 2 (1) ◽  
pp. 8
Author(s):  
Ronak Bakhtiari ◽  
Jalil Fallah Mehrabadi ◽  
Hedroosha Molla Agamirzaei ◽  
Ailar Sabbaghi ◽  
Mohammad Mehdi Soltan Dallal

Resistance to b-lactam antibiotics by gramnegative bacteria, especially <em>Escherichia coli (E. coli)</em>, is a major public health issue worldwide. The predominant resistance mechanism in gram negative bacteria particularly <em>E. coli </em>is via the production of extended spectrum beta lactamase (ESBLs) enzymes. In recent years, the prevalence of b-lactamase producing organisms is increased and identification of these isolates by using disk diffusion method and no-one else is not satisfactory. So, this investigation focused on evaluating the prevalence of ESBL enzymes by disk diffusion method and confirmatory test (Combined Disk). Five hundred clinical samples were collected and 200 <em>E. coli </em>isolates were detected by standard biochemical tests. To performing initial screening of ESBLs was used from Disk diffusion method on <em>E. coli </em>isolates. A confirmation test (Combined Disk method) was performed on isolates of resistant to cephalosporin's indicators. Up to 70% isolates exhibited the Multi Drug Resistance phenotype. In Disk diffusion method, 128(64%) <em>E. coli </em>isolates which resistant to ceftazidime and cefotaxime while in Combined Disk, among 128 screened isolates, 115 (89.8%) isolates were detected as ESBLs producers. This survey indicate beta lactamase enzymes are playing a significant role in antibiotic resistance and correct detection of them in phenotypic test by using disk diffusion and combined Disk is essential for accurate recognition of ESBLs.


2007 ◽  
Vol 59 (2) ◽  
pp. 508-512 ◽  
Author(s):  
B.R. Paneto ◽  
R.P. Schocken-Iturrino ◽  
C. Macedo ◽  
E. Santo ◽  
J.M. Marin

The occurrence of toxigenic Escherichia coli in raw milk cheese was surveyed in Middle Western Brazil. Fifty samples of cheese from different supermarkets were analyzed for E.coli. The isolates were serotyped and screened for the presence of verotoxigenic E. coli (VTEC) and enterotoxigenic E. coli (ETEC) by Polymerase Chain Reaction (PCR). The susceptibility to thirteen antimicrobial agents was evaluated by the disk diffusion method. E.coli were recovered from 48 (96.0%) of the samples. The serogroups identified were O125 (6.0%), O111 (4.0%), O55 (2.0%) and O119 (2.0%). Three (6.0%) and 1(2.0%) of the E.coli isolates were VTEC and ETEC, respectively. Most frequent resistance was observed to the following antimicrobials: cephalothin (60.0%), nalidixic acid (40.0%), doxycyclin (33.0%), tetracycline (31.0%) and ampicillin (29.0%).


2020 ◽  
Vol 4 (3) ◽  
pp. 323-327
Author(s):  
Mamunu Abdulkadir SULAIMAN ◽  
H.S Muhammad ◽  
Aliyu Muhammad Sani ◽  
Aminu Ibrahim ◽  
Ibrahim Muhammad Hussain ◽  
...  

Multidrug resistance (MDR) exhibited by some strains of Escherichia coli may be due to acquiring mobile genetic element (R-plasmid) by the bacteria, or intrinsically induced by inappropriate use of antibiotics by the hosts.  Infection by such strains may result to prolonged illness and greater risk of death. The study evaluated the impact of curing on antibiotic resistance on selected clinical isolates of E. coli. Twenty clinical isolates of E. coli from our previous studies were re-characterized using conventional microbiological techniques. Antibiotic sensitivity testing was determined by disk diffusion method, MDR selected based on resistance to ≥ 2 classes of antibiotics. Multiple antibiotic resistance (MAR) index was determined as ratio of the number of antibiotic resisted to the total number of antibiotics tested and considered significant if ≥. 0.2. The isolates that showed significant MAR index were subjected to plasmid curing using acridine orange, thereafter, profiled for plasmid and the cured ones were re-tested against the antibiotics they initially resisted. Out of the 20 isolates, 19 (95%) were confirmed as E. coli, all (100%) of which were MDRs, which was highest against augmentin (78.9%) followed by amoxacillin (52.6%). However, after the plasmid curing only 6 (31.6%) out of the 19 isolates cured retained significant MAR index and the level of the significance had reduced drastically in 16 (84.2%) isolates. Conclusively, curing assay can completely eliminate R-plasmid acquired resistance. More studied on plasmid curing agents for possible augmentation of the agents into antibiotics may see the rise of successful antibiotic era again.


2021 ◽  
Vol 14 (8) ◽  
Author(s):  
Seyed Ali Bazghandi ◽  
Mohsen Arzanlou ◽  
Hadi Peeridogaheh ◽  
Hamid Vaez ◽  
Amirhossein Sahebkar ◽  
...  

Background: Drug resistance and virulence genes are two key factors for the colonization of Pseudomonas aeruginosa in settings with high antibiotic pressure, such as hospitals, and the development of hospital-acquired infections. Objectives: The objective of this study was to investigate the prevalence of drug resistance and virulence gene profiles in clinical isolates of P. aeruginosa in Ardabil, Iran. Methods: A total of 84 P. aeruginosa isolates were collected from clinical specimens of Ardabil hospitals and confirmed using laboratory standard tests. The disk diffusion method was used for antibiotic susceptibility testing and polymerase chain reaction (PCR) for the identification of P. aeruginosa virulence genes. Results: The highest and the lowest antibiotic resistance rates of P. aeruginosa strains were against ticarcillin-clavulanate (94%) and doripenem (33.3%), respectively. In addition, the frequency of multidrug-resistant (MDR) P. aeruginosa was 55.9%. The prevalence of virulence factor genes was as follows: algD 84.5%, lasB 86.9%, plcH 86.9%, plcN 86.9%, exoU 56%, exoS 51.2%, toxA 81%, nan1 13.1%, and pilB 33.3%. A significant association was observed between resistance to some antibiotics and the prevalence of virulence genes in P. aeruginosa. Conclusions: Our results revealed a high prevalence of antibiotic resistance, especially MDR, and virulence-associated genes in clinical isolates of P. aeruginosa in Ardabil hospitals. Owing to the low resistance rates against doripenem, gentamicin, and tobramycin, these antibiotics are recommended for the treatment of infections caused by highly resistant and virulent P. aeruginosa strains.


Animals ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 396 ◽  
Author(s):  
Michaela Sannettha van den Honert ◽  
Pieter Andries Gouws ◽  
Louwrens Christiaan Hoffman

Studies have shown that antibiotic resistance among wild animals is becoming a public health concern, owing to increased contact and co-habitation with domestic animals that, in turn, results in increased human contact, indirectly and directly. This type of farming practice intensifies the likelihood of antibiotic resistant traits in microorganisms transferring between ecosystems which are linked via various transfer vectors, such as rivers and birds. This study aimed to determine whether the practice of wildlife supplementary feeding could have an influence on the antibiotic resistance of the bacteria harboured by the supplementary fed wildlife, and thus play a potential role in the dissemination of antibiotic resistance throughout nature. Escherichia coli and Enterococcus were isolated from the faeces of various wildlife species from seven different farms across South Africa. The Kirby-Bauer disk diffusion method was used according to the Clinical and Laboratory Standards Institute 2018 guidelines. The E. coli (F: 57%; N = 75% susceptible) and Enterococcus (F: 67%; N = 78% susceptible) isolates from the supplementary fed (F) wildlife were in general, found to be more frequently resistant to the selection of antibiotics than from those which were not supplementary fed (N), particularly towards tetracycline (E. coli F: 56%; N: 71%/Enterococcus F: 53%; N: 89% susceptible), ampicillin (F: 82%; N = 95% susceptible) and sulphafurazole (F: 68%; N = 98% susceptible). Interestingly, high resistance towards streptomycin was observed in the bacteria from both the supplementary fed (7% susceptible) and non-supplementary fed (6% susceptible) wildlife isolates. No resistance was found towards chloramphenicol and ceftazidime.


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