Antimicrobial resistance patterns and phylogenetic analysis of Shiga toxin-producing Escherichia coli strains from goats using both Clermont phylogenetic schemes

Foodborne transmission of Shiga toxin-producing Escherichia coli (STEC) poses a threat to public health. The Clermont typing schemes (previous and revised) have been used widely to phylotype E. coli. The present study was conducted to compare the relationship of the Clermont phylogenet-ic schemes in STEC strains isolated from goats and antibiotic resistance patterns in the southeast of Iran. Overall 52 strains carrying the stx gene were used for subsequent analysis. All strains were determined by analysing the genomic DNA with a PCR-based method using the two Clermont et al. (2000, 2013) schemes. Extended spectrum beta-lactamase (ESBL) producing strains were con-firmed by the double disk-diffusion method. STEC strains were also tested for susceptibility to 20 antimicrobials agents. In the original Clermont method, the prevalent phylogroups were B1 (69.2%) and A (28.8%). The significant phylogenetic groups of strains according to the revised Clermont method were B1 (82.7%), A (13.5%) and unknown (3.8%). However, STEC strains underwent changes as noted from A to B1 (17.3%), B1 to unknown (3.8%), B1 to A (1.9%) and D to B1 (1.9%) groupings. Of the 52 stx-positive strains, two ESBL producing strains were detected. Susceptibility data showed that the most frequent resistance phenotype was related to cefazolin (90.4%), streptomycin (88.5%), ampicillin (86.5%) and oxytetracycline (82.7%) respectively. Alt-hough the overall frequency of the reassigned phylotypes was not significant, most changes oc-curred within the A phylotype. Therefore, implementation of the new method on isolates belong-ing to the A phylotype in the old method seems to be necessary to obtain accurate results.

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The frequency of virulent genes and antimicrobial resistance patterns of diarrheagenic Escherichia coli isolated from stools of children presenting with diarrhea in a tertiary hospital in Abakaliki, Nigeria

International Journal of One Health ◽

10.14202/ijoh.2020.147-152 ◽

2020 ◽

Vol 6 (2) ◽

pp. 147-152

Author(s):

Ebuka Elijah David ◽

Muhammad Arfat Yameen ◽

Ikechuku Okorie Igwenyi ◽

Arthur Chinedu Okafor ◽

Uket Nta Obeten ◽

...

Keyword(s):

Escherichia Coli ◽

Tertiary Care ◽

Tertiary Hospital ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Diarrheagenic Escherichia Coli ◽

Resistance Patterns ◽

Etiological Agents ◽

Antibiotic Resistance Patterns

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.

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Prevalence of Shiga Toxin-Producing Escherichia coli O157 and Non-O157 Serogroups Isolated from Fresh Raw Beef Meat Samples in an Industrial Slaughterhouse

International Journal of Microbiology ◽

10.1155/2021/1978952 ◽

2021 ◽

Vol 2021 ◽

pp. 1-6

Author(s):

Kiandokht Babolhavaeji ◽

Leili Shokoohizadeh ◽

Morteza Yavari ◽

Abbas Moradi ◽

Mohammad Yousef Alikhani

Keyword(s):

Escherichia Coli ◽

Shiga Toxin ◽

Genetic Linkage ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Antibiotic Resistant ◽

E Coli ◽

Beef Meat ◽

Resistance Patterns ◽

Meat Samples

Background. The aims of the current study are the identification of O157 and non-O157 Shiga Toxin-Producing Escherichia coli (STEC) serogroups isolated from fresh raw beef meat samples in an industrial slaughterhouse, determination of antimicrobial resistance patterns, and genetic linkage of STEC isolates. Materials and Methods. A total of 110 beef samples were collected from the depth of the rump of cattle slaughtered at Hamadan industrial slaughterhouse. After detection of E. coli isolates, STEC strains were identified according to PCR for stx1, stx2, eaeA, and hlyA virulence genes, and STEC serogroups (O157 and non-O157) were identified by PCR. The genetic linkage of STEC isolates was analyzed by the ERIC- (Enterobacterial Repetitive Intergenic Consensus-) PCR method. The antimicrobial susceptibility of STEC isolates was detected by the disk diffusion method according to CLSI guidelines. Results. Among 110 collected beef samples, 77 (70%) were positive for E. coli. The prevalence of STEC in E. coli isolates was 8 (10.4%). The overall prevalence of O157 and non-O157 STEC isolates was 12.5% (one isolate) and 87.5% (7 isolates), respectively. The hemolysin gene was detected in 25% (2 isolates) of STEC strains. Evaluation of antibiotic resistance indicated that 100% of STEC isolates were resistant to ampicillin, ampicillin-sulbactam, amoxicillin-clavulanic acid, and cefazolin. Resistance to tetracycline and ciprofloxacin was detected in 62.5% and 12.5% of isolates, respectively. The analysis of the ERIC-PCR results showed five different ERIC types among the STEC isolates. Conclusion. The isolation of different clones STECs from beef and the presence of antibiotic-resistant isolates indicate that more attention should be paid to the hygiene of slaughterhouses.

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Prevalence of Shiga toxin-producing and Enteropathogenic Escherichia coli Isolated from Chicken Meat in the west of Iran

10.21203/rs.2.11448/v4 ◽

2019 ◽

Author(s):

omid zarei ◽

Leili Shokoohizadeh ◽

Hadi Hossainpour ◽

Mohammad Yousef Alikhani

Keyword(s):

Escherichia Coli ◽

Shiga Toxin ◽

Genetic Relatedness ◽

Foodborne Pathogen ◽

Chicken Meat ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Resistance Patterns ◽

High Level

Abstract Objective: Shiga toxin-producing Escherichia coli (STEC) is known as a crucial zoonotic foodborne pathogen. Totally, 257 raw chicken meat were collected from markets in Hamadan, west of Iran. The samples were cultured on selective media and the virulence genes of E. coli isolates were analyzed by PCR. The antibiotic resistance patterns were determined by the disk diffusion method. The genetic relatedness of the E. coli O157 isolates was analyzed by ERIC-PCR. Results: Totally, 93 (36%; 95% CI 41.9- 30.1%) isolates were identified as E. coli. Based on microbiological tests, 36 (38.7%; 95% CI 48.6-28.8), 7 (7.5%; 95% CI 12.8-2.2%), and 12 (12.9%; 95% CI 19.7- 6.1%) of the E. coli isolates were characterized as STEC, Enteropathogenic E. coli, and attaching and effacing E. coli (AEEC) strains, respectively. A high level of resistance to nalidixic acid (91.4%; 95% CI 97.1- 85.7%), tetracycline (89.8%; 95% CI 96.2-83.5%), ampicillin (82.8%; 95% CI 90.2-75.1%), and sulfametoxazole-trimotoprime (71%; 95% CI 80.2-61.8%) was detected among the E. coli isolates. The analysis of ERIC-PCR results showed five different ERIC types among the E. coli O157 isolates. Based on findings. Control and check-up of poultry meats should be considered as a crucial issue for public health.

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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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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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Antibiotic Resistance Patterns of Diverse Escherichia coli Phylogenetic Groups Isolated from the Al-Hillah River in Babylon Province, Iraq

The Scientific World JOURNAL ◽

10.1155/2019/5927059 ◽

2019 ◽

Vol 2019 ◽

pp. 1-8 ◽

Cited By ~ 2

Author(s):

Mourouge Saadi Alwash ◽

Hawraa Mohammed Al-Rafyai

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Anthropogenic Activities ◽

Health Concern ◽

Resistant Bacteria ◽

Anthropogenic Pressures ◽

Phylogenetic Groups ◽

E Coli ◽

Resistance Patterns ◽

Antibiotic Resistance Patterns

Surface water contamination remains a major worldwide public health concern and may contribute to the dissemination of antibiotic-resistant bacteria. The Al-Hillah River in the city of Babylon Province, Iraq, diverts flows from the Euphrates River. Because of its importance in irrigation and population density, it faces several forced and unforced changes due to anthropogenic activities. To evaluate water quality, water samples were collected from three sites with different anthropogenic pressures along the Al-Hillah River. These samples were subjected to bacteriological analyses, i.e., total coliforms, Escherichia coli, and faecal enterococci. The phylogenetic groups of the E. coli isolates (n = 61) were typed by rapid PCR-based analyses. Representatives of each isolate were tested phenotypically for resistance to six classes of antibiotics and characterized according to their phylogenetic groups. The results demonstrated the highest resistance levels were to β-lactam antibiotics, followed by fosfomycin and aminoglycosides. Escherichia coli isolates belonging to phylogenetic groups A and B2 were the most common and were characterized by a higher prevalence of antibiotic resistance. This study is important for understanding the current conditions of the Al-Hillah River, as the data reveal a high prevalence of multiresistance among E. coli isolates circulating at the three sampling sites.

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Drug resistance, serotypes, and phylogenetic groups among uropathogenic Escherichia coli including O25-ST131 in Mexico City

The Journal of Infection in Developing Countries ◽

10.3855/jidc.1703 ◽

2011 ◽

Vol 5 (12) ◽

pp. 840-849 ◽

Cited By ~ 36

Author(s):

José Molina-López ◽

Gerardo Aparicio-Ozores ◽

Rosa María Ribas-Aparicio ◽

Sandra Gavilanes-Parra ◽

María Elena Chávez-Berrocal ◽

...

Keyword(s):

Escherichia Coli ◽

Multidrug Resistance ◽

Mexico City ◽

Uropathogenic Escherichia Coli ◽

Diffusion Method ◽

Phylogenetic Groups ◽

Disk Diffusion Method ◽

E Coli ◽

Pcr Multiplex ◽

Tract Infections

Introduction: The increasing prevalence of uropathogenic Escherichia coli (UPEC) strains resistant to multiple antibiotics complicates the treatment of urinary tract infections (UTIs). This study aimed to analyze the antimicrobial resistance, serotypes, and phylogenetic groups among strains of E. coli isolated from outpatients with UTIs in Mexico City. Methodology: A total of 119 E. coli isolates were recovered from urine samples from outpatients with clinical diagnosis of uncomplicated UTIs from 2004 to 2007. The serotype was assessed by agglutination in microtiter plates; susceptibility to antimicrobials was determined by the disk diffusion method. Clone O25-ST131 and phylogenetic groups of E. coli strains were tested by methods based on PCR multiplex. Results: The predominant serotype was O25:H4 (21.2%). Resistance to antibiotics was ampicillin (83.7%); piperacillin (53.8%); the fluoroquinolone group (55.5-60.6%), and trimethoprim/sulfamethoxazole (TMP/SMX) (56.4%). Additionally, 36 (30.2%) isolates were multidrug-resistant and 13 of these 36 strains were identified as E. coli O25-ST131 clone by an allele-specific PCR-based assay. Phylogenetic analysis showed that 15 of 17 isolates with serotype O25:H4 belonged to group B2. Conclusions: This is the first report that establishes the presence in Mexico of the O25-ST131 clonal group of E. coli, which has been associated with multidrug-resistance and with high virulence potential. The spread of this clone in Mexico should be monitored closely. We found a correlation between serotype O25:H4 and multidrug resistance in UPEC strains. Our results indicate that the use of ampicillin, fluoroquinolones, and TMP/SMX should be reviewed when selecting empirical therapy for UTIs.

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Antimicrobial Resistance in Outpatient Escherichia coli Urinary Isolates in Dakar, Senegal

The Journal of Infection in Developing Countries ◽

10.3855/jidc.362 ◽

2007 ◽

Vol 1 (03) ◽

pp. 263-268 ◽

Cited By ~ 20

Author(s):

Jean-Marie Sire ◽

Pierre Nabeth ◽

Jean-David Perrier-Gros-Claude ◽

Ibrahim Bahsoun ◽

Tidiane Siby ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Antimicrobial Susceptibility ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Extended Spectrum Beta Lactamase ◽

Predominant Pathogen ◽

Overall Resistance ◽

Tract Infections ◽

Resistance Rates

Background: Data regarding the evolution of antimicrobial resistance are needed to suggest appropriate empirical treatment of urinary tract infections (UTI) in developing countries. To assess the antimicrobial susceptibility of Escherichia coli, the predominant pathogen in community-acquired UTI, a prospective multicenter study was carried out in Dakar, Senegal. Methodology: From February 2004 to October 2006, 1010 non-duplicate E. coli strains were collected from four centres. Antimicrobial susceptibility testing was performed using disk diffusion method according to the recommendations of the CA-SFM (2004). Results: Most of the isolates were resistant to amoxicillin (73.1%), amoxicillin-clavulanic acid (67.5%), cephalothin (55.8%), and trimethoprim/sulfamethoxazole (68.1%). Extended spectrum beta-lactamase was detected in 38 strains. The overall resistance rates to nalidixic acid, norfloxacin and ciprofloxacin were 23.9%, 16.4% and 15.5%, respectively. Most of the strains were susceptible to gentamicin, nitrofurantoin and fosfomycin (respective susceptibility rates, 93.8%, 89.9%, and 99.3%). During this period, a significant decrease in sensitivity was observed for cephalothin, fluoroquinolones and trimethoprim/sulfamethoxazole (p

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Pathogenic features of urinary Escherichia coli strains causing Asymptomatic Bacteriuria during Pregnancy

10.21203/rs.2.22234/v1 ◽

2020 ◽

Author(s):

Samane Mohebi ◽

zahra Hashemizade ◽

Mahtab Hadadi ◽

Soudeh Kholdi ◽

Kasra Javadi ◽

...

Keyword(s):

Escherichia Coli ◽

Virulence Genes ◽

Asymptomatic Bacteriuria ◽

Urine Samples ◽

Diffusion Method ◽

Low Birth Weight Infants ◽

Phylogenetic Groups ◽

Disk Diffusion Method ◽

Cross Sectional ◽

E Coli

Abstract Background Asymptomatic bacteriuria is one of the common problems in pregnancy. Pyelonephritis, preterm labor and low birth weight infants have been associated with bacterial infection. Urinary tract infection (UTI) during pregnancy is frequently associated with complications. An observational cross-sectional study including investigated the prevalence of virulence genes, antimicrobial resistance, and its relationship with phylogenetic groups among E. coli strains isolated from pregnant women with asymptomatic bacteriuria who referred to Hafez hospital, Shiraz, Iran.Material and Methods A total of 300 urine samples were screened for Escherichia coli strains. Susceptibility testing was determined by the disk-diffusion method. The phylogenetic groups and 13 virulence genes were identified by PCR. ESBL and AmpC producing isolates were detected using phenotypic methods. PCR was used to identify the bla TEM , bla SHV and bla CTXM genes in ESBL and AmpC-positive isolates.Results Our results revealed that among 300 urine samples, 105 (35%) were positive for E. coli . The data showed that the highest and the lowest resistance rates were observed against nalidixic acid (82.1%), and imipenem (2.8%), respectively. The prevalence of ESBLs and AmpC-β-lactamase, in the E. coli isolates was 41% and 9.5% respectively. bla CTXM was the commonest genotype (93%). Phylogenetic group distribution was as follow: B1 2.8%, A 14.2%, B2 61.9%, and D 4.6%. Our result showed that most of the virulence genes belonged to group B2 and also several virulence genes such as hlyA , cnf-1 , and papGII genes were positively associated with group B2. Conclusion Among E. coli strains isolated from patients with UTIs, different features phylogroups, with special virulence factors, could cause severe infection. Awareness about the Virulence patterns distribution among Phylogenetic groups of UPEC could greatly aid in confine and prevent the development of lethal infection caused by these strains.

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Prevalence and antibiotic resistance profile of Shiga toxin-producing Escherichia coli isolated from diarrheal samples

Iranian Journal of Microbiology ◽

10.18502/ijm.v12i4.3931 ◽

2020 ◽

Author(s):

Farzad Esavand Heydari ◽

Mojtaba Bonyadian ◽

Hamdallah Moshtaghi ◽

Masoud Sami

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Shiga Toxin ◽

Virulence Genes ◽

Enterohemorrhagic Escherichia Coli ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Resistance Profile ◽

E Coli ◽

Antibiotic Resistance Profile

Background and Objectives: Enterohemorrhagic Escherichia coli (EHEC) causes bloody and non-bloody diarrhea, intestinal infection and extraintestinal complications in humans. This study aimed to isolate and evaluate the prevalence of E. coli O157: H7 and other Shiga toxin-producing E. coli (STEC) and identify the virulence genes (stx1, stx2, hly and eaeA) from patients with diarrhea. Also, the antibiotic resistance profile of the isolated strains was evaluated. Materials and Methods: A total of 100 stool samples were collected from patients with acute diarrhea referring to the hospital and clinics in Isfahan County, Iran. Phenotypic tests and PCR assay were used for detection of E. coli O157: H7 and other Shiga toxin-producing E. coli. The presence of virulence genes (stx1, stx2, hly and eaeA) were identified by PCR. The antibiotic resistance profile of the isolates was determined using the agar disk diffusion method. The results were analyzed descriptively by Sigma stat version 4 software. Results: Seventy - eight out of 100 samples (78%) were contaminated with E. coli. E. coli O157 was isolated from five samples (6.4%), of which only two strains (2.56%) were identified as E. coli O157: H7. According to the results, out of two E. coli O157: H7 isolates, one (50%) isolate contained eaeA and two isolates (100%) contained Stx1, Stx2, hlyA genes. Out of three (3.84%) E. coli O157: HN, one of the isolate (33.3%) contained stx1 and, two isolates (66.7%) were positive for hlyA genes. Also, the results revealed that six strains (7.69%) were non-O157: H7 STEC, of which two isolates (33.3%) contained stx1 and four isolates (66.7%) were positive for stx2 and hlyA genes. The results of antibiogram tests revealed that all of the STEC isolates (100%) were sensitive to imipenem followed by kanamycin, gentamicin and nitrofurantoin (91%). High resistance (54.5%) to ampicillin and ciprofloxacin was observed among the STEC isolates. Conclusion: The results of the current study showed that although the prevalence of E. coli O157: H7 was low among patients with diarrhea, the other STEC strains with relative resistance to antibiotics are more prevalent.

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