Prevalence and Molecular Characterization of Antimicrobial Resistance in Escherichia coli Isolated from Raw Milk and Raw Milk Cheese in Egypt

ABSTRACT The goal of this study was to examine antimicrobial resistance and characterize the implicated genes in 222 isolates of Escherichia coli from 187 samples of raw milk and the two most popular cheeses in Egypt. E. coli isolates were tested for susceptibility to 12 antimicrobials by a disk diffusion method. Among the 222 E. coli isolates, 66 (29.7%) were resistant to one or more antimicrobials, and half of these resistant isolates showed a multidrug resistance phenotype (resistance to at least three different drug classes). The resistance traits were observed to tetracycline (27.5%), ampicillin (18.9%), streptomycin (18.5%), sulfamethoxazole-trimethoprim (11.3%), cefotaxime (4.5%), kanamycin (4.1%), ceftazidime (3.6%), chloramphenicol (2.3%), nalidixic acid (1.8%), and ciprofloxacin (1.4%). No resistance to fosfomycin and imipenem was observed. Tetracycline resistance genes tetA, tetB, and tetD were detected in 53 isolates, 9 isolates, and 1 isolate, respectively, but tetC was not detected. Aminoglycoside resistance genes strA, strB, aadA, and aphA1 were detected in 41, 41, 11, and 9 isolates, respectively. Sulfonamide resistance genes sul1, sul2, and sul3 were detected in 7, 25, and 3 isolates, respectively. Of 42 ampicillin-resistant isolates, blaTEM, blaCTX-M, and blaSHV were detected in 40, 9, and 3 isolates, respectively, and 10 (23.8%) ampicillin-resistant isolates were found to produce extended-spectrum β-lactamase. Each bla gene of extended-spectrum β-lactamase–producing E. coli was further subtyped to be blaCTX-M-15, blaCTX-M-104, blaTEM-1, and blaSHV-12. The class 1 integron was also detected in 28 resistant isolates, and three different patterns were obtained by PCR-restriction fragment length polymorphism. Sequencing analysis of the variable region revealed that four isolates had dfrA12/orfF/aadA2, two had aadA22, and one had dfrA1/aadA1. These data suggest that antimicrobial-resistant E. coli are widely distributed in the milk production and processing environment in Egypt and may play a role in dissemination of antimicrobial resistance to other pathogenic and commensal bacteria.

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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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Antimicrobial-Resistant and Extended-Spectrum β-Lactamase–Producing Escherichia coli in Raw Cow's Milk

Journal of Food Protection ◽

10.4315/0362-028x.jfp-14-250 ◽

2015 ◽

Vol 78 (1) ◽

pp. 72-77 ◽

Cited By ~ 21

Author(s):

ALENA SKOČKOVÁ ◽

KATEŘINA BOGDANOVIČOVÁ ◽

IVANA KOLÁČKOVÁ ◽

RENÁTA KARPÍŠKOVÁ

Keyword(s):

Escherichia Coli ◽

Resistance Genes ◽

Diffusion Method ◽

Cow’S Milk ◽

Public Health Issue ◽

Resistant Bacteria ◽

Cow's Milk ◽

E Coli ◽

Extended Spectrum ◽

Raw Cow’S Milk

The occurrence of antimicrobial-resistant bacteria is an important public health issue. The aim of this study was the monitoring of resistant Escherichia coli in raw cow's milk with a focus on the detection of extended-spectrum β-lactamase (ESBL)–producing strains. In total, 263 samples of raw milk from 40 farms were collected and investigated in 2010 to 2013 in the Czech Republic. Detection of E. coli was performed and evaluated according to ISO 16649-2, and antibiotic resistance was screened by the disk diffusion method. The presence of E. coli was detected in 243 (92.4%) samples. In total, 270 isolates were obtained. Resistance to β-lactam (31.8%) and tetracycline (13.0%) antibiotics was detected most often and also multiresistant strains (5.5%) were observed. E. coli isolates found to be resistant to β-lactam, tetracycline, and quinolone antibiotics were assayed by PCR to detect selected genes encoding those resistance mechanisms. In isolates in which any bla genes were detected, a double-disk synergy test was performed. ESBL production was confirmed in 2 (0.7%) isolates. The genetic analysis identified the presence of the blaCTX-M gene and other resistance genes (tet(B) and qnrB). Both ESBL-positive isolates originated from the same farm and had an identical pulsed-field gel electrophoresis profile. The findings of our study indicate that milk can be a reservoir of bacteria carrying resistance genes with a potential for spreading through the food chain.

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Fecal carriage of extended-spectrum β-lactamase-producing Enterobacteriaceae in hospital and community settings in Chad

Antimicrobial Resistance and Infection Control ◽

10.1186/s13756-019-0626-z ◽

2019 ◽

Vol 8 (1) ◽

Cited By ~ 3

Author(s):

Oumar Ouchar Mahamat ◽

Abdelsalam Tidjani ◽

Manon Lounnas ◽

Mallorie Hide ◽

Julio Benavides ◽

...

Keyword(s):

Escherichia Coli ◽

Pathogenic Bacteria ◽

Hospitalized Patients ◽

Diffusion Method ◽

Community Settings ◽

Carriage Rate ◽

Disk Diffusion Method ◽

E Coli ◽

Fecal Carriage ◽

Extended Spectrum

Abstract Background Fecal carriage of extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-PE) remains poorly documented in Africa. The objective of this study was to determine the prevalence of ESBL-PE fecal carriage in Chad. Methods In total, 200 fresh stool samples were collected from 100 healthy community volunteers and 100 hospitalized patients from January to March 2017. After screening using ESBL-selective agar plates and species identification by MALDI-TOF mass spectrometry, antibiotic susceptibility was tested using the disk diffusion method, and ESBL production confirmed with the double-disc synergy test. The different ESBL genes in potential ESBL-producing isolates were detected by PCR and double stranded DNA sequencing. Escherichia coli phylogenetic groups were determined using a PCR-based method. Results ESBL-PE fecal carriage prevalence was 44.5% (51% among hospitalized patients vs 38% among healthy volunteers; p < 0.05). ESBL-producing isolates were mostly Escherichia coli (64/89) and Klebsiella pneumoniae (16/89). PCR and sequencing showed that 98.8% (87/89) of ESBL-PE harbored blaCTX-M genes: blaCTX-M-15 in 94.25% (82/87) and blaCTX-M-14 in 5.75% (5/87). Phylogroup determination by quadruplex PCR indicated that ESBL-producing E. coli isolates belonged to group A (n = 17; 27%), C (n = 17; 27%), B2 (n = 9; 14%), B1 (n = 8; 13%), D (n = 8; 13%), E (n = 1; 1.6%), and F (n = 1; 1.6%). The ST131 clone was identified in 100% (9/9) of E. coli B2 strains. Conclusions The high fecal carriage rate of ESBL-PE associated with CTX-M-15 in hospital and community settings of Chad highlights the risk for resistance transmission between non-pathogenic and pathogenic bacteria.

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Molecular Characterization of β-Lactamase Producing Genes and Integrons in Diarrheagenic Escherichia Coli From Diarrheal Children Less Than Five Years of Age in Ouagadougou, Burkina Faso.

10.21203/rs.3.rs-628039/v1 ◽

2021 ◽

Author(s):

Rene Dembele ◽

Wendpoulomdé A.D. Kaboré ◽

Issiaka Soulama ◽

Oumar Traoré ◽

Nafissatou Ouédraogo ◽

...

Keyword(s):

Escherichia Coli ◽

Molecular Characterization ◽

Resistance Genes ◽

Antibiotic Resistance Genes ◽

Control Measures ◽

Diffusion Method ◽

Biochemical Tests ◽

Disk Diffusion Method ◽

E Coli

Abstract Background The aim of this study was to determine the resistance of diarrheagenic Escherichia coli strains to β-lactams antibiotics and to perform the molecular characterization of Extended Spectrum β-lactamases (ESBL) and integrons genes. Methods This study was carried out from August 2013 to October 2015 and involved 31 DEC strains isolated from diarrheal stools samples collected from children less than five years of age. The identification and characterization of DEC strains was done through the standard biochemical tests those were confirmed using API 20E and Polymerase Chain Reaction (PCR). The determination of antimicrobial resistance was realized by the disk diffusion method then an amplification of the β-lactamase resistance genes and integrons by PCR was done. Results Out of the 419 E. coli strains identified, 31 isolates (7.4%) harbored the DEC virulence genes. From these DEC, 21 (67.7%) were ESBL-producing E. coli. Susceptibility to ESBL-producing E. coli showed that the majority of isolates were highly resistant to amoxicillin (77.4%), amoxicillin clavulanic acid (77.4%) and piperacillin (64.5%). The following antibiotic resistance genes and integron were identified from the 31 DEC isolates: blaTEM (6.5%), blaSHV (19.4%), blaOXA (38.7%) blaCTX−M (9.7%), Int1 (58.1%) and Int3 (19.4%). No class 2 integrons (Int2) was characterized. Conclusions Because of the high prevalence of multidrug-resistant ESBL organisms found in this study among pediatric patients, there is a need of stringent pediatric infection control measures.

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Antibiotic Resistance of Escherichia coli Isolated from Processing of Brewery Waste with the Addition of Bulking Agents

Sustainability ◽

10.3390/su131810174 ◽

2021 ◽

Vol 13 (18) ◽

pp. 10174

Author(s):

Katarzyna Wolny-Koładka ◽

Marek Zdaniewicz

Keyword(s):

Escherichia Coli ◽

Drug Resistance ◽

Antibiotic Resistance ◽

Diffusion Method ◽

Bulking Agents ◽

Disk Diffusion Method ◽

Beta Lactamases ◽

E Coli ◽

Extended Spectrum ◽

Extended Spectrum Beta Lactamases

The aim of the study was to determine the drug resistance profile and to assess the presence of genes responsible for the production of extended-spectrum beta-lactamases in Escherichia coli isolated from energy-processed hop sediment with the addition of bulking agents. Antibiotic resistance was determined by the disk diffusion method and the PCR technique to detect genes determining the extended-spectrum beta-lactamases (ESBLs) mechanism. A total of 100 strains of E. coli were collected. The highest resistance was found to aztreonam, tetracycline, ampicillin, ticarcillin, and ceftazidime. The bacteria collected were most often resistant to even 10 antibiotics at the same time and 15 MDR strains were found. The ESBL mechanism was determined in 14 isolates. Among the studied genes responsible for beta-lactamase production, blaTEM was the most common (64%). The study revealed that the analysed material was colonised by multi-drug-resistant strains of E. coli, which pose a threat to public health. The obtained results encourage further studies to monitor the spread of drug resistance in E. coli.

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Multiple and High-Risk Clones of Extended-Spectrum Cephalosporin-Resistant and blaNDM-5-Harbouring Uropathogenic Escherichia coli from Cats and Dogs in Thailand

Antibiotics ◽

10.3390/antibiotics10111374 ◽

2021 ◽

Vol 10 (11) ◽

pp. 1374

Author(s):

Naiyaphat Nittayasut ◽

Jitrapa Yindee ◽

Pongthai Boonkham ◽

Teerapong Yata ◽

Nipattra Suanpairintr ◽

...

Keyword(s):

Escherichia Coli ◽

High Risk ◽

Antimicrobial Resistance ◽

Resistance Genes ◽

Multidrug Resistant ◽

Integrase Gene ◽

E Coli ◽

Extended Spectrum ◽

Antimicrobial Resistance Genes ◽

Tract Infections

Resistance to extended-spectrum cephalosporins (ESC) and carbapenems in Escherichia coli (E. coli), increasingly identified in small animals, indicates a crisis of an antimicrobial resistance situation in veterinary medicine and public health. This study aimed to characterise the genetic features of ESC-resistant E. coli isolated from cats and dogs with urinary tract infections in Thailand. Of 72 ESC-resistant E. coli isolated from diagnostic samples (2016–2018), blaCTX-M including group 1 (CTX-M-55, -15 and -173) and group 9 (CTX-M-14, -27, -65 and -90) variants were detected in 47 isolates (65.28%) using PCR and DNA sequencing. Additional antimicrobial resistance genes, including plasmid-mediated AmpC (CIT and DHA), blaNDM-5, mcr-3, mph(A) and aac(6′)-Ib-cr, were detected in these isolates. Using a broth microdilution assay, all the strains exhibited multidrug-resistant phenotypes. The phylogroups were F (36.11%), A (20.83%), B1 (19.44%), B2 (19.44%) and D (4.17%), with several virulence genes, plasmid replicons and an integrase gene. The DNA fingerprinting using a repetitive extragenic palindromic sequence-PCR presented clonal relationships within phylogroups. Multiple human-associated, high-risk ExPEC clones associated with multidrug resistance, including sequence type (ST) 38, ST131, ST224, ST167, ST354, ST410, ST617 and ST648, were identified, suggesting clonal dissemination. Dogs and cats are a potential reservoir of ESC-resistant E. coli and significant antimicrobial resistance genes.

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Occurrence of Escherichia coli producing extended spectrum β-lactamases in food-producing animals

Acta Veterinaria Hungarica ◽

10.1556/004.2021.00036 ◽

2021 ◽

Author(s):

Bence Balázs ◽

József Bálint Nagy ◽

Zoltán Tóth ◽

Fruzsina Nagy ◽

Sándor Károlyi ◽

...

Keyword(s):

Escherichia Coli ◽

Gene Families ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Chain Reactions ◽

E Coli ◽

Esbl Gene ◽

Source Of Infection ◽

Extended Spectrum ◽

Esbl Producers

Abstract Multidrug resistance due to the production of extended-spectrum beta-lactamases (ESBLs) is a major problem in human as well as in veterinary medicine. These strains appear in animal and human microbiomes and can be the source of infection both in animal and in human healthcare, in accordance with the One Health theorem. In this study we examined the prevalence of ESBL-producing bacteria in food-producing animals. We collected 100 porcine and 114 poultry samples to examine the prevalence of ESBL producers. Isolates were identified using the MALDI-TOF system and their antibiotic susceptibility was tested using the disk diffusion method. ESBL gene families and phylogroups were detected by polymerase chain reactions. The prevalence of ESBL producers was relatively high in both sample groups: 72 (72.0%) porcine and 39 (34.2%) poultry isolates were ESBL producers. Escherichia coli isolates were chosen for further investigations. The most common ESBL gene was CTX-M-1 (79.3%). Most of the isolates belong to the commensal E. coli phylogroups. The porcine isolates could be divided into three phylogroups, while the distribution of the poultry isolates was more varied. In summary, ESBL-producing bacteria are prevalent in the faecal samples of the examined food-producing animals, with a dominance of the CTX-M-1 group enzymes and commensal E. coli phylogroups.

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Antibiotic resistance and extended-spectrum β-lactamase in Escherichia coli isolates from imported 1-day-old chicks, ducklings, and turkey poults

Veterinary World ◽

10.14202/vetworld.2020.1037-1044 ◽

2020 ◽

Vol 13 (6) ◽

pp. 1037-1044

Author(s):

Mona A. A. AbdelRahman ◽

Heba Roshdy ◽

Abdelhafez H. Samir ◽

Engy A. Hamed

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Diffusion Method ◽

Poultry Production ◽

Disk Diffusion Method ◽

Antibiotic Resistant ◽

E Coli ◽

Potential Source ◽

Extended Spectrum ◽

Turkey Poults

Aim: Antimicrobial resistance is a global health threat. This study investigated the prevalence of Escherichia coli in imported 1-day-old chicks, ducklings, and turkey poults. Materials and Methods: The liver, heart, lungs, and yolk sacs of 148 imported batches of 1-day-old flocks (chicks, 45; ducklings, 63; and turkey poults, 40) were bacteriologically examined for the presence of E. coli. Results: We isolated 38 E. coli strains from 13.5%, 6.7%, and 5.4% of imported batches of 1-day-old chicks, ducklings, and turkey poults, respectively. They were serotyped as O91, O125, O145, O78, O44, O36, O169, O124, O15, O26, and untyped in the imported chicks; O91, O119, O145, O15, O169, and untyped in the imported ducklings; and O78, O28, O29, O168, O125, O158, and O115 in the imported turkey poults. The E. coli isolates were investigated for antibiotic resistance against 16 antibiotics using the disk diffusion method and were found resistant to cefotaxime (60.5%), nalidixic acid (44.7%), tetracycline (44.7%), and trimethoprim-sulfamethoxazole (42.1%). The distribution of extended-spectrum β-lactamase (ESBL) and ampC β-lactamase genes was blaTEM (52.6%), blaSHV (28.9%), blaCTX-M (39.5%), blaOXA-1 (13.1%), and ampC (28.9%). Conclusion: Imported 1-day-old poultry flocks may be a potential source for the dissemination of antibiotic-resistant E. coli and the ESBL genes in poultry production.

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