Molecular types, virulence profiles and antimicrobial resistance of Escherichia coli causing bovine mastitis

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.

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

Baghdad Science Journal ◽

10.21123/bsj.2019.16.3(suppl.).0682 ◽

2019 ◽

Vol 16 (3(Suppl.)) ◽

pp. 0682 ◽

Cited By ~ 1

Author(s):

MKK Et al.

Keyword(s):

Escherichia Coli ◽

Clinical Specimen ◽

Pcr Amplification ◽

Tertiary Hospital ◽

Clinical Isolates ◽

Diffusion Method ◽

Clinical Samples ◽

Disk Diffusion Method ◽

E Coli ◽

Resistance Rates

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.

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Antibiotic Resistance of Escherichia Coli Isolated from Broiler Chickens

Folia Veterinaria ◽

10.2478/fv-2019-0021 ◽

2019 ◽

Vol 63 (3) ◽

pp. 1-8

Author(s):

M. Sciberras ◽

M. Pipová ◽

I. Regecová ◽

P. Jevinová ◽

S. Demjanová

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Broiler Chickens ◽

Tetracycline Resistance ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Phenotypic Resistance ◽

E Coli ◽

Mueller Hinton Agar ◽

Almost All

Abstract The purpose of this study was to detect the antibiotic resistance of forty-one Escherichia coli isolates from the intestinal contents of slaughtered broiler chickens using the disk diffusion method according to Kirby-Bauer. Mueller-Hinton agar plates were inoculated with 0.1 ml overnight broth cultures of individual E. coli isolates and the disks with the following concentrations of antibiotics were applied onto them: ampicillin (10 μg), cefotaxime (30 μg), gentamicin (10 μg), streptomycin (10 μg), azithromycin (15 μg), tetracycline (30 μg), ciprofloxacin (30 μg) and levofloxacin (3 μg). After the incubation at 37 °C for 16—18 hours, the inhibition zones were measured and interpreted in accordance with the Clinical and Laboratory Standard Institute (CLSI) zone diameter breakpoints. Almost all E. coli isolates showed resistance to tetracycline (92.68 %), most of them were resistant to gentamicin (75.61 %) and levofloxacine (70.73 %). Phenotypic resistance to tetracycline was further confirmed with the help of the Polymerase Chain Reaction (PCR) procedure focused on the presence of specific tet(A) and tet(B) genes. These genes were detected in all 41 E. coli isolates. On the contrary, E. coli isolates were highly susceptible to both azithromycin and streptomycin. In conclusion, the study highlighted the role of commensal E. coli bacteria isolated from the intestines of broiler chickens as an important reservoir of tetracycline resistance genes.

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Escherichia coli Isolated from Urinary Tract Infections of Lebanese Patients between 2000 and 2009: Epidemiology and Profiles of Resistance

Chemotherapy Research and Practice ◽

10.1155/2011/218431 ◽

2011 ◽

Vol 2011 ◽

pp. 1-6 ◽

Cited By ~ 10

Author(s):

Ziad Daoud ◽

Claude Afif

Keyword(s):

Escherichia Coli ◽

Urinary Tract ◽

Urinary Tract Infections ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Standard Culture ◽

Tract Infections ◽

Susceptibility Profiles ◽

Third Generation Cephalosporins

The purpose of this study was to investigate the bacterial etiology of urinary tract infections in one of the busiest hospitals of Lebanon and to examine the epidemiologic and microbiologic properties of Escherichia coli isolated from urinary tract infections of Lebanese patients over a 10-year period. Methods. This retrospective study analyzed the data generated between 2000 and 2009 (10,013 Gram-positive and Gram-negative bacteria). Bacterial identification was based on standard culture and biochemical characteristics of isolates. Antimicrobial susceptibility was tested by the disk diffusion method, and ESBL production was detected by synergy with third-generation cephalosporins and amoxiclav. Results. E. coli was the most frequent isolate throughout the ten years (60.64% of the total isolates). It was followed by Klebsiella pneumoniae and Proteus sp., Pseudomonas aeruginosa, Enterococcus sp., and Streptococcus agalactiae. E. coli occurred more frequently in women (69.8%) than in men (61.4%). The lowest percentage of susceptibility of E. coli was manifested against piperacillin and ampicillin. An increase in the production of ESBL was observed (2.3% in 2000 to 16.8% in 2009). Conclusions. The etiology of urinary tract infections and their susceptibility profiles are important to be evaluated in countries like Lebanon where a severe misuse of antibiotics at all levels is observed.

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Molecular characterization of antimicrobial resistance and virulence genes of Escherichia coli isolates from bovine mastitis

Veterinary World ◽

10.14202/vetworld.2020.1588-1593 ◽

2020 ◽

Vol 13 (8) ◽

pp. 1588-1593

Author(s):

Zuhair Bani Ismail ◽

Sameeh M. Abutarbush

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Shiga Toxin ◽

Virulence Genes ◽

Bovine Mastitis ◽

Virulence Gene ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Procaine Penicillin

Background and Aim: Mastitis is a common and economically important disease in dairy cattle. It remains one of the most common reasons for the extensive use of antimicrobials in dairy farms leading to the emergence of antimicrobial-resistant pathogens. The aim of this study was to determine the patterns of antimicrobial resistance of Escherichia coli isolates from bovine mastitis and to identify prominent antimicrobial resistance and virulence genes among isolated strains. Materials and Methods: Antimicrobial susceptibility testing against six antibiotic groups, including tetracyclines, aminoglycosides, beta-lactams, macrolides, sulfonamides, and fluoroquinolones was performed using the disk diffusion method. PCR was performed on resistant isolates to detect resistance and virulence genes using commercially available primers. Results: Out of 216 milk samples cultured, 14 samples yielded E. coli isolates. All isolates (100%) were resistant to ampicillin, amoxicillin, procaine penicillin, streptomycin, oxytetracycline, and sulfamethoxazole-trimethoprim. Only one isolate (7%) was sensitive to gentamicin, and all isolates (100%) were sensitive to enrofloxacin and ciprofloxacin. All isolates carried at least one resistance gene against one or more of the major antibiotic groups. All isolates carried the ereA, tetG, tetE, and tetB genes, followed by tetA (93%), ampC (86%), strA (86%), sul1 (78%), tetD (71%), tetC (57%), aadA (57%), and strB (36%). The lowest percentage of isolates carried bla1 (17%) and bla2 (12%) genes, and none of the isolates carried the qnrA gene. Most of the isolates (93%) carried the Shiga toxin 1 virulence gene, followed by complement resistance protein (79%), intimin (64%), Shiga toxin 2 (36%), cytotoxic necrotizing factor (35%), aerotaxis receptor (21%), and type 1 fimbriae (15%). Conclusion: Results of this study indicate that the high percentages of E. coli isolate from bovine mastitis are resistant to two or more of the major antibiotic groups, irrespective of the presence or absence of relevant resistance or virulence genes.

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Molecular Characterization of Carbapenemase-Producing Enterobacterales in Children with Diarrhea in Rural Burkina Faso

Journal of Drug Delivery and Therapeutics ◽

10.22270/jddt.v11i1.4513 ◽

2021 ◽

Vol 11 (1) ◽

pp. 84-92

Author(s):

René Dembélé ◽

Issiaka Soulama ◽

Wendpoulomdé Aimé Désiré Kaboré ◽

Ali Konaté ◽

Assèta Kagambèga ◽

...

Keyword(s):

Escherichia Coli ◽

Burkina Faso ◽

Treatment Options ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Klebsiella Pneumoniae Carbapenemase ◽

Resistance Rates ◽

Encoding Genes

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

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Phenotypic Antimicrobial Susceptibility of Escherichia coli from Raw Meats, Ready-to-Eat Meats, and Their Related Samples in One Health Context

Microorganisms ◽

10.3390/microorganisms9020326 ◽

2021 ◽

Vol 9 (2) ◽

pp. 326

Author(s):

Frederick Adzitey ◽

Nurul Huda ◽

Amir Husni Mohd Shariff

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Antimicrobial Susceptibility ◽

Resistance Index ◽

Diffusion Method ◽

Resistant Bacteria ◽

Disk Diffusion Method ◽

E Coli ◽

The Usa ◽

Raw Meats

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.

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Evaluation of extended spectrum beta lactamase enzymes prevalence in clinical isolates of Escherichia coli

Microbiology Research ◽

10.4081/mr.2011.e8 ◽

2011 ◽

Vol 2 (1) ◽

pp. 8

Author(s):

Ronak Bakhtiari ◽

Jalil Fallah Mehrabadi ◽

Hedroosha Molla Agamirzaei ◽

Ailar Sabbaghi ◽

Mohammad Mehdi Soltan Dallal

Keyword(s):

Escherichia Coli ◽

Disk Diffusion ◽

Confirmatory Test ◽

Diffusion Method ◽

Multi Drug Resistance ◽

Beta Lactamase ◽

Disk Diffusion Method ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Extended Spectrum

Resistance to b-lactam antibiotics by gramnegative bacteria, especially Escherichia coli (E. coli), is a major public health issue worldwide. The predominant resistance mechanism in gram negative bacteria particularly E. coli 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 E. coli isolates were detected by standard biochemical tests. To performing initial screening of ESBLs was used from Disk diffusion method on E. coli 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%) E. coli 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.

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Occurrence of toxigenic Escherichia coli in raw milk cheese in Brazil

Arquivo Brasileiro de Medicina Veterinária e Zootecnia ◽

10.1590/s0102-09352007000200035 ◽

2007 ◽

Vol 59 (2) ◽

pp. 508-512 ◽

Cited By ~ 27

Author(s):

B.R. Paneto ◽

R.P. Schocken-Iturrino ◽

C. Macedo ◽

E. Santo ◽

J.M. Marin

Keyword(s):

Escherichia Coli ◽

Polymerase Chain Reaction ◽

Antimicrobial Agents ◽

Raw Milk ◽

Diffusion Method ◽

Chain Reaction ◽

Disk Diffusion Method ◽

E Coli ◽

Polymerase Chain ◽

Raw Milk Cheese

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%).

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Survey and Sequence Characterization of Bovine Mastitis-Associated Escherichia coli in Dairy Herds

Frontiers in Veterinary Science ◽

10.3389/fvets.2020.582297 ◽

2020 ◽

Vol 7 ◽

Author(s):

John I. Alawneh ◽

Ben Vezina ◽

Hena R. Ramay ◽

Hulayyil Al-Harbi ◽

Ameh S. James ◽

...

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Bovine Mastitis ◽

Antibiotic Resistance Genes ◽

Tetracycline Resistance ◽

Sequence Type ◽

Genotypic Differences ◽

Beta Lactam ◽

Sequence Characterization ◽

E Coli

Escherichia coli is frequently associated with mastitis in cattle. “Pathogenic” and “commensal” isolates appear to be genetically similar. With a few exceptions, no notable genotypic differences have been found between commensal and mastitis-associated E. coli. In this study, 24 E. coli strains were isolated from dairy cows with clinical mastitis in three geographic regions of Australia (North Queensland, South Queensland, and Victoria), sequenced, then genomically surveyed. There was no observed relationship between sequence type (ST) and region (p = 0.51). The most common Multi Locus Sequence Type was ST10 (38%), then ST4429 (13%). Pangenomic analysis revealed a soft-core genome of 3,463 genes, including genes associated with antibiotic resistance, chemotaxis, motility, adhesion, biofilm formation, and pili. A total of 36 different plasmids were identified and generally found to have local distributions (p = 0.02). Only 2 plasmids contained antibiotic resistance genes, a p1303_5-like plasmid encoding multidrug-resistance (trimethoprim, quaternary ammonium, beta-lactam, streptomycin, sulfonamide, and kanamycin) from two North Queensland isolates on the same farm, while three Victorian isolates from the same farm contained a pCFSAN004177P_01-like plasmid encoding tetracycline-resistance. This pattern is consistent with a local spread of antibiotic resistance through plasmids of bovine mastitis cases. Notably, co-occurrence of plasmids containing virulence factors/antibiotic resistance with putative mobilization was rare, though the multidrug resistant p1303_5-like plasmid was predicted to be conjugative and is of some concern. This survey has provided greater understanding of antibiotic resistance within E. coli-associated bovine mastitis which will allow greater prediction and improved decision making in disease management.

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A Preliminary Study: Antibiotic Resistance Patterns of Escherichia coli and Enterococcus Species from Wildlife Species Subjected to Supplementary Feeding on Various South African Farms

Animals ◽

10.3390/ani10030396 ◽

2020 ◽

Vol 10 (3) ◽

pp. 396 ◽

Cited By ~ 1

Author(s):

Michaela Sannettha van den Honert ◽

Pieter Andries Gouws ◽

Louwrens Christiaan Hoffman

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Supplementary Feeding ◽

Health Concern ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Wildlife Species ◽

Human Contact ◽

Resistance Patterns

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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