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

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.

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Detection of CMY-type beta-lactamases in Escherichia coli isolates from paediatric patients in a tertiary care hospital in Mexico

Antimicrobial Resistance and Infection Control ◽

10.1186/s13756-020-00840-4 ◽

2020 ◽

Vol 9 (1) ◽

Author(s):

Jocelin Merida-Vieyra ◽

Agustín De Colsa-Ranero ◽

Yair Calderón-Castañeda ◽

Alejandra Aquino-Andrade

Keyword(s):

Boronic Acid ◽

Tertiary Care ◽

Fourth Generation ◽

Production Test ◽

Care Hospital ◽

Disk Diffusion Test ◽

Beta Lactamases ◽

E Coli ◽

Paediatric Patients ◽

Extended Spectrum Beta Lactamases

Abstract Background The aim of this study was to detect CMY-type beta-lactamases in E. coli isolates obtained from paediatric patients. Methods In total, 404 infection-causing E. coli isolates resistant to third and fourth generation cephalosporins (3GC, 4GC) were collected from paediatric patients over a 2 years period. The identification and susceptibility profiles were determined with an automated microbiology system. Typing of blaCMY and other beta-lactamase genes (blaTEM, blaSHV, blaCTX-M, blaVIM, blaIMP, blaKPC, blaNDM, blaOXA and blaGES) was realized by PCR and sequencing. Phenotypic detection of AmpC-type enzymes was performed using boronic acid (20 mg/mL) and cloxacillin (20 mg/mL) as inhibitors, and the production of extended-spectrum beta-lactamases was determined with the double-disk diffusion test with cefotaxime (CTX) and ceftazidime (CAZ) discs alone and in combination with clavulanic acid. The CarbaNP test and modified carbapenem inhibition method (mCIM) were used for isolates with decreased susceptibility to carbapenems. The clonal origin of the isolates was established by pulsed-field gel electrophoresis (PFGE), phylotyping method and multilocus sequence typing. Results CMY-type beta-lactamases were detected in 18 isolates (4.5%). The allelic variants found were CMY-2 (n = 14) and CMY-42 (n = 4). Of the E. coli strains with CMY, the AmpC phenotypic production test was positive in 11 isolates with cloxacillin and in 15 with boronic acid. ESBL production was detected in 13 isolates. Coexistence with other beta-lactamases was observed such as CTX-M-15 ESBL and original spectrum beta-lactamases TEM-1 and TEM-190. In one isolate, the CarbaNP test was negative, the mCIM was positive, and OXA-48 carbapenemase was detected. Phylogroup A was the most frequent (n = 9) followed by B2, E and F (n = 2, respectively), and through PFGE, no clonal relationship was observed. Eleven different sequence types (ST) were found, with ST10 high-risk clone being the most frequent (n = 4). Seventy-two percent of the isolates were from health care-associated infections; the mortality rate was 11.1%. Conclusions This is the first report in Mexico of E. coli producing CMY isolated from paediatric patients, demonstrating a frequency of 4.5%. In addition, this is the first finding of E. coli ST10 with CMY-2 and OXA-48.

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Paper-based analytical devices for colorimetric detection of S. aureus and E. coli and their antibiotic resistant strains in milk

The Analyst ◽

10.1039/d0an01075h ◽

2020 ◽

Vol 145 (22) ◽

pp. 7320-7329

Author(s):

Muhammad Asif ◽

Fazli Rabbi Awan ◽

Qaiser Mahmood Khan ◽

Bongkot Ngamsom ◽

Nicole Pamme

Keyword(s):

Pathogenic Bacteria ◽

Colorimetric Detection ◽

Antibiotic Resistant ◽

Beta Lactamases ◽

E Coli ◽

Milk Samples ◽

Extended Spectrum Beta Lactamases ◽

Paper Microfluidic ◽

Resistant Strains ◽

Appropriate Prescription

We investigate paper microfluidic devices for detection of pathogenic bacteria and their sensitivity towards β-lactamase and Extended Spectrum Beta Lactamases (ESBLs) in milk samples to enable appropriate prescription of antibiotics for mastitis.

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Antibiotic resistance and virulence of Escherichia coli strains isolated from animal rendering plant

Scientific Reports ◽

10.1038/s41598-020-72851-5 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Gabriela Gregova ◽

Vladimir Kmet

Keyword(s):

Escherichia Coli ◽

Antibacterial Agents ◽

Virulence Genes ◽

Gene Encoding ◽

Antibiotic Resistant ◽

Beta Lactamases ◽

E Coli ◽

Animal Wastes ◽

Extended Spectrum Beta Lactamases ◽

High Level

Abstract Processing of animal carcasses and other animal wastes in rendering plants is a significant source of antibiotic resistant microorganisms. The main goal of this study was to investigate the resistance to 18 antibacterial agents including β-lactams, fluoroquinolones, colistin and virulence factors (iss, tsh, cvaC, iutA, papC, kps and ibeA genes) in 88 Escherichia coli strains isolated from a rendering plant over 1 year period. ESBL (Extended-spectrum beta-lactamases) and plasmid-mediated Amp were screened by interpretative reading of MIC. ESBL phenotype was detected in 20.4% of samples and high level of resistance to fluoroquinolone was found in 27.2% of strains. Cephalosporinase CTX-M1, cephamycinase CMY-2, integrase 1 and transposon 3 genes were detected by PCR. Furthermore, there were found three CMY-2 producing E. coli with O25b-ST131, resistant to the high level of enrofloxacin and containing the gene encoding the ferric aerobactin receptor (iutA). One enrofloxacin resistant E. coli strain possessed iss, ibeA, kps and papC virulence genes also with CMY-2, integrase1 and Tn3. ST131 E. coli with CMY-2 has a zoonotic potential and presents a serious health risk to humans.

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Single Ribosomal Protein Mutations in Antibiotic-Resistant Bacteria Analyzed by Mass Spectrometry

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.45.11.3046-3055.2001 ◽

2001 ◽

Vol 45 (11) ◽

pp. 3046-3055 ◽

Cited By ~ 28

Author(s):

Sheri K. Wilcox ◽

Gregory S. Cavey ◽

James D. Pearson

Keyword(s):

Mass Spectrometry ◽

Antibiotic Resistance ◽

Ribosomal Proteins ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

Mutant Proteins ◽

E Coli ◽

Maldi Tof ◽

Molecular Masses

ABSTRACT Mutations in several ribosomal proteins are known to be related to antibiotic resistance. For several strains of Escherichia coli, the mutated protein is known but the amino acid actually altered has not been documented. Characterization of these determinants for antibiotic resistance in proteins will further the understanding of the precise mechanism of the antibiotic action as well as provide markers for resistance. Mass spectrometry can be used as a valuable tool to rapidly locate and characterize mutant proteins by using a small amount of material. We have used electrospray and matrix-assisted laser desorption ionization–time of flight (MALDI–TOF) mass spectrometry to map out all 56 ribosomal proteins in E. coli based on intact molecular masses. We used this fingerprinting approach to locate variants of ribosomal proteins displaying a change in mass. In particular we have studied proteins responsible for streptomycin, erythromycin, and spectinomycin resistance in three strains of E. coli, and then we characterized each mutation responsible for resistance by analyzing tryptic peptides of these proteins by using MALDI-TOF and nanoelectrospray tandem mass spectrometry. The results provided markers for antibiotic resistance and demonstrated that mass spectrometry can be used to rapidly investigate changes in individual proteins from a complex with picomole amounts of protein.

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Community Origin and Previous Use of Antimicrobials Increase the Risk of Nosocomial Multidrug-resistant Bacteria Colonisation in the Intensive Care Unit in a Brazilian Hospital

The Open Public Health Journal ◽

10.2174/1874944501912010449 ◽

2019 ◽

Vol 12 (1) ◽

pp. 449-454

Author(s):

Debora Vandresen ◽

Maria Helena B. Werlang ◽

Mirian Carla B. Silva ◽

Juliana S. Link ◽

Paulo Cezar N. Fortes

Keyword(s):

Intensive Care Unit ◽

Intensive Care ◽

Antibiotic Use ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Antibiotic Resistant ◽

Beta Lactamases ◽

Hospitalised Patients ◽

Extended Spectrum Beta Lactamases ◽

Hospital Acquired

Background: Hospitalised patients are often surrounded by microorganisms, and antibiotic-resistant pathogens are a major and growing threat to public health. Objective: This study aimed to investigate the epidemiology and the risk factors for colonisation by multidrug-resistant organisms (MDROs) in a Brazilian hospital. Methods: Patients in the Intensive Care Unit (ICU) who underwent nasal and rectal swab cultures for the surveillance of colonisation by MDROs were evaluated in a retrospective study. MDROs were determined by routine microbiological cultures. Results: Of the 785 patients included in this study, 86 presented positive results for MDRO colonisation. Overall, the most frequently isolated organism was Klebsiella pneumoniae (41.9%), followed by Escherichia coli (33.7%). The main type of resistance was the production of extended-spectrum beta-lactamases (ESBL). The prevalence of MDRO infections was significantly associated with the patient's origin (community or hospital-acquired). Having been submitted to previous antimicrobial drug therapy was significantly associated with MDRO infection (relative risk [RR]: 4.02 [2.60 - 6.23]). Conclusion: MDRO ICU colonisation was variable, with similar frequencies as other centres, and important factors, including previous hospital stay and antibiotic use, were closely related to MDRO colonisation. Therefore, control interventions should reduce their rates, especially considering the particularities of each geographic centre.

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

10.21203/rs.3.pex-1588/v1 ◽

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.

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Occurrence of extended-spectrum beta-lactamases in human and bovine isolates of Escherichia coli from Oyo state, Nigeria

The Journal of Infection in Developing Countries ◽

10.3855/jidc.3430 ◽

2014 ◽

Vol 8 (06) ◽

pp. 774-779 ◽

Cited By ~ 16

Author(s):

Christiana Inwezerua ◽

Nuno Mendonça ◽

Vera Calhau ◽

Sara Domingues ◽

Olufemi Ezekiel Adeleke ◽

...

Keyword(s):

Escherichia Coli ◽

Recipient Strain ◽

Beta Lactamases ◽

E Coli ◽

Class 1 Integrons ◽

Extended Spectrum ◽

Plasmid Replicon ◽

Class 1 ◽

Extended Spectrum Beta Lactamases

Introduction: The main objective of the study was the molecular characterization of extended spectrum β-lactamases (ESBL) in Escherichia coli isolates collected from human and bovine samples in Oyo state, Nigeria. Methodology: Between August 2010-2011, 114 E. coli isolates were collected from hospitals (n = 57) and bovine (n = 57). PCR and sequencing were used for identification of ESBLs, upstream sequences, plasmid-mediated quinolone resistance (PMQR) genes and class 1 integrons. Plasmid incompatibility groups were identified among ESBL-positive isolates by PCR. Genetic relatdness was assessed by rep-PCR and MLST. Transfer of ESBL determinants to the recipient strain E. coli J53 was performed by broth mating assays. Results: CTX-M15 was the unique ESBL found in eight human isolates. Six CTX-M-15 producers also carry the aac(6’)-lb-cr gene and/or qnrB gene, and class 1 integrons. FIA, FIB, H11, H12, F, Y and K were the plasmid replicon types found. CTX-M-15 and PMQR determinants were transferred by conjugation in two E. coli assigned by MLST to ST131 and ST2695, a new allele. Conclusions: The study highlights the dissemination hability of CTX-M-15 associated with PMQR, and the presence of class 1 integrons, able to capture additional genes, justifying the urgent need of antimicrobial resistance surveillance in Nigeria.

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A trimethoprim derivative impedes antibiotic resistance evolution

Nature Communications ◽

10.1038/s41467-021-23191-z ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Madhu Sudan Manna ◽

Yusuf Talha Tamer ◽

Ilona Gaszek ◽

Nicole Poulides ◽

Ayesha Ahmed ◽

...

Keyword(s):

Antibiotic Resistance ◽

Resistant Bacteria ◽

Gene Encoding ◽

Antibiotic Resistant ◽

E Coli ◽

Resistance Evolution ◽

Evolutionary Trajectories ◽

Evolution Of Resistance

AbstractThe antibiotic trimethoprim (TMP) is used to treat a variety of Escherichia coli infections, but its efficacy is limited by the rapid emergence of TMP-resistant bacteria. Previous laboratory evolution experiments have identified resistance-conferring mutations in the gene encoding the TMP target, bacterial dihydrofolate reductase (DHFR), in particular mutation L28R. Here, we show that 4’-desmethyltrimethoprim (4’-DTMP) inhibits both DHFR and its L28R variant, and selects against the emergence of TMP-resistant bacteria that carry the L28R mutation in laboratory experiments. Furthermore, antibiotic-sensitive E. coli populations acquire antibiotic resistance at a substantially slower rate when grown in the presence of 4’-DTMP than in the presence of TMP. We find that 4’-DTMP impedes evolution of resistance by selecting against resistant genotypes with the L28R mutation and diverting genetic trajectories to other resistance-conferring DHFR mutations with catalytic deficiencies. Our results demonstrate how a detailed characterization of resistance-conferring mutations in a target enzyme can help identify potential drugs against antibiotic-resistant bacteria, which may ultimately increase long-term efficacy of antimicrobial therapies by modulating evolutionary trajectories that lead to resistance.

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Occurrence of bacteria with a dangerous extent of antibiotic resistance in poultry in the Central Region of Moravia

Acta Veterinaria Brno ◽

10.2754/avb201887020165 ◽

2018 ◽

Vol 87 (2) ◽

pp. 165-172 ◽

Cited By ~ 2

Author(s):

Jan Bardoň ◽

Patrik Mlynárčik ◽

Petra Procházková ◽

Magdaléna Röderová ◽

Kristýna Mezerová ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Broad Spectrum ◽

Clinical Samples ◽

Resistant Bacteria ◽

Methicillin Resistant ◽

Beta Lactamases ◽

E Coli ◽

Human Food Chain ◽

Extended Spectrum Beta Lactamases ◽

Phenotypic Tests

The study aimed to determine the occurrence ofEnterobacteriaceaeproducing broad-spectrum beta-lactamases, vancomycin-resistant enterococci (VRE) and methicillin-resistantStaphylococcus aureusstrains in poultry in Moravia, Czech Republic, including phenotypic and genotypic analyses of the extent of resistance. Using chromogenic screening media, a total of 240 clinical samples collected from poultry and the poultry farm environment were processed. Phenotypic tests identified 23 isolates of broad-spectrum beta-lactamase-producingEscherichia coliand one VRE isolate (Enterococcus faeciumwith VanA resistance). Methicillin-resistantStaphylococcus aureusstrains were not detected. Among the isolates producing broad-spectrum beta-lactamases, 17 produced extended-spectrum beta-lactamases, most frequently CTX-M; the remaining 6 isolates were CIT-type AmpC enzymes. No carbapenemase-producing strains were detected. Pulsed-field gel electrophoresis showed that 21E. colistrains (91%) were genetically unrelated isolates. Increasing resistance of bacteria to antibiotic agents poses a serious issue for both human and veterinary medicine globally. For humans, a potential source of resistant bacteria may be animals or their products entering the human food chain, for example poultry. The presented study extends existing knowledge about the occurrence of resistant bacteria in poultry in Moravia and describes the phenotype and genotype of their resistance to antibiotics.

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

Antibiotics ◽

10.3390/antibiotics10050510 ◽

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.

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