scholarly journals Bacteriological and Molecular Characterization of Diarrheagenic Escherichia Coli Pathotypes From Children in Duhok City, Iraq

2020 ◽  
Vol 8 (2) ◽  
pp. 52-57
Author(s):  
Haveen K. Hasan ◽  
Najim A. Yassin ◽  
Souzan H. Eassa
Keyword(s):  
Artificial Feeding ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Different Types ◽  
Resistance Patterns ◽  
Vitek 2 ◽  
Antibiotic Resistance Patterns ◽  
Susceptibility Profiles

Background: A variety of diarrheagenic E coli (DEC) are responsible for causing different types of diarrhea in children especially in developing courtiers. Objectives: This study was primarily aimed to isolate and bacteriological characterizing of E coli from diarrheic infant stool and to investigate their antibiotic resistance patterns and then using molecular identification of DEC pathotypes for better discrimination. Methods: Total of 400 fresh stools specimens were collected from children with diarrhea in Heevi Hospital in Duhok city, Iraq. The samples were cultured on selective media such as (MacConkey and MacConkey sorbitol agar). Colonies were identified through biochemical reaction and VITEK 2 system and then antibiotic susceptibility profiles were determined. Results: A total of 349(87.2%) samples were yielded positive for growth of E coli. Out of these, 50 phenotypically-identified E coli were then subjected to PCR assay targeting certain virulence factors (alt, eae, sxt1 and sxt2) for discrimination of pathotyes. 13/50(26%) Enterotoxigenix E. coli (ETEC) was detected, 5/50(10%) Enterohemorrhagic E coli EHEC was detected, while no Enteropathogenic E coli (EPEC) was detected. All pathotypes were more frequent in samples from male children aged between 2-3 years that were artificial feeding pattern. Moreover, all pathotypes expressed high resistant to ampicillin, cephalosporin and tetracycline while little resistance to imipenem was observed. Conclusion: The study concludes presence of diarrheagenic E coli pathotypes in our community causing diarrhea in children and emphasize on using of PCR assay for best discrimination. 

scholarly journals 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

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.

scholarly journals ESBL Producing Escherichia coli in Faecal Sludge Treatment Plants: An Invisible Threat to Public Health in Rohingya Camps, Cox's Bazar, Bangladesh

2021 ◽  
Vol 9 ◽  
Author(s):  
Md. Sakib Hossain ◽  
Sobur Ali ◽  
Monir Hossain ◽  
Salman Zahir Uddin ◽  
M. Moniruzzaman ◽  
...  
Keyword(s):  
Public Health ◽  
Virulence Gene ◽  
Multidrug Resistant ◽  
Sludge Treatment ◽  
E Coli ◽  
Resistance Patterns ◽  
Plasmid Profiling ◽  
Antibiotic Resistance Patterns ◽  
Faecal Sludge

Introduction: Human faecal sludge contains diverse harmful microorganisms, making it hazardous to the environment and public health if it is discharged untreated. Faecal sludge is one of the major sources of E. coli that can produce extended-spectrum β-lactamases (ESBLs).Objective: This study aimed to investigate the prevalence and molecular characterization of ESBL-producing E. coli in faecal sludge samples collected from faecal sludge treatment plants (FSTPs) in Rohingya camps, Bangladesh.Methods: ESBL producing E. coli were screened by cultural as well as molecular methods and further characterized for their major ESBL genes, plasmid profiles, pathotypes, antibiotic resistance patterns, conjugation ability, and genetic similarity.Results: Of 296 isolates, 180 were phenotypically positive for ESBL. All the isolates, except one, contained at least one ESBL gene that was tested (blaCTX−M−1, blaCTX−M−2, blaCTX−M−8, blaCTX−M−9, blaCTX−M−15, blaCTX−M−25, blaTEM, and blaSHV). From plasmid profiling, it was observed that plasmids of 1–211 MDa were found in 84% (151/180) of the isolates. Besides, 13% (24/180) of the isolates possessed diarrhoeagenic virulence genes. From the remaining isolates, around 51% (79/156) harbored at least one virulence gene that is associated with the extraintestinal pathogenicity of E. coli. Moreover, 4% (3/156) of the isolates were detected to be potential extraintestinal pathogenic E. coli (ExPEC) strains. Additionally, all the diarrhoeagenic and ExPEC strains showed resistance to three or more antibiotic groups which indicate their multidrug-resistant potential. ERIC-PCR differentiated these pathogenic isolates into seven clusters. In addition to this, 16 out of 35 tested isolates transferred plasmids of 32–112 MDa to E. coli J53 recipient strain.Conclusion: The present study implies that the faecal sludge samples examined here could be a potential origin for spreading MDR pathogenic ESBL-producing E. coli. The exposure of Rohingya individuals, living in overcrowded camps, to these organisms poses a severe threat to their health.

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

scholarly journals Characterization of cefotaxime resistant Escherichia coli isolated from broiler farms in Ecuador

2018 ◽  
Author(s):  
Christian Vinueza-Burgos ◽  
David Ortega-Paredes ◽  
Cristian Narváez ◽  
Lieven De Zutter ◽  
Jeannete Zurita
Keyword(s):  
Antimicrobial Resistance ◽  
Latin American ◽  
Production Systems ◽  
Diffusion Method ◽  
Poultry Production ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Resistance Patterns ◽  
Latin American Countries

AbstractAntimicrobial resistance (AR) is a worldwide concern. Up to a 160% increase in antibiotic usage in food animals is expected in Latin American countries. The poultry industry is an increasingly important segment of food production and contributor to AR. The objective of this study was to evaluate the prevalence, AR patterns and the characterization of relevant resistance genes in Extended Spectrum β-lactamases (ESBL) and AmpC E. coli from large poultry farms in Ecuador. Sampling was performed from June 2013 to July 2014 in 6 slaughterhouses that slaughter broilers from 115 farms totaling 384 flocks. Each sample of collected caeca was streaked onto TBX agar supplemented with cefotaxime (3 mg/l). In total, 176 isolates were analyzed for antimicrobial resistance patterns by the disk diffusion method and for blaCTX-M, blaTEM, blaCMY, blaSHV, blaKPC, and mcr-1 by PCR and sequencing. ESBL and AmpC E. coli were found in 362 flocks (94.3%) from 112 farms (97.4%). We found that 98.3% of the isolates were multi-resistant to antibiotics. Low resistance was observed for ertapenem and nitrofurantoin. The most prevalent ESBL genes were the blaCTX-M (90.9%) blaCTX-M-65, blaCTX-M-55 and blaCTX-M-3 alleles. Most of the AmpC strains presented the blaCMY-2 gene. Three isolates showed the mcr-1 gene. Poultry production systems represent a hotspot for antimicrobial resistance in Ecuador, possibly mediated by the extensive use of antibiotics. Monitoring this sector in national and regional plans of antimicrobial resistance surveillance should therefore be considered.

scholarly journals Molecular characterization of extended-spectrum beta-lactamase producing Enterobacteriaceae in a Saudi Arabian tertiary hospital

2014 ◽  
Vol 8 (03) ◽  
pp. 282-288 ◽  
Author(s):  
Hoda Hassan ◽  
Baha Abdalhamid
Keyword(s):  
Tertiary Hospital ◽  
Multidrug Resistant ◽  
Beta Lactamase ◽  
M Gene ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Extended Spectrum ◽  
Vitek 2 ◽  
High Level

Introduction: The aim of this study was to determine the prevalence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae), and Proteus mirabilis (P. mirabilis). In addition, different methods for detection of these enzymes, including the newly introduced CHROMagar ESBL, were evaluated. Methodology: A total of 382 Enterobacteriaceae clinical isolates were obtained from King Fahad Specialist Hospital – Dammam, during 2011 and screened for production of ESBL using advanced expert system of Vitek 2, CHROMagar and ESBL-E-strips. PCR assay was used to detect blaTEM, blaSHV, and blaCTX-M genes. Susceptibility to a panel of antibiotics was determined. Results: The overall proportion of ESBL-producing enterobacterial isolates was 30.6%, which was higher in E. coli (35.8%) than in K. pneumoniae (25.7%). ESBL genotypes showed remarkable increase in the CTX-M (97.4%) compared to SHV (23.1%). The predominant ESBL was CTX-M- 15 (92.1 %). No TEM ESBL was detected in this study. The Vitek2 showed the highest sensitivity (100%), and the CHROMagar had the lowest specificity (97.3%) compared to the molecular method. All isolates were susceptible to imipenem and meropenem. Conclusions: This study confirms a high level of blaCTX-M positive ESBL isolates are circulating in the Eastern Province of Saudi Arabia. The trend of a multidrug-resistant profile associated with the recovery of the blaCTX-M gene is alarming.

scholarly journals Molecular Detection of Diarrheagenic Escherichia coli from Children with Acute Diarrhea in Tertiary Care Hospitals of Dhaka, Bangladesh.

2013 ◽  
Vol 5 (2) ◽  
pp. 59-66 ◽  
Author(s):  
Sushmita Roy ◽  
SM Shamsuzzaman ◽  
Kazi Z Mamun
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Acute Diarrhea ◽  
Tertiary Care ◽  
Medical Science ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Stool Samples

Objective: Multiplex PCR assay was used for diagnosis of diarrheagenic Escherichia coli (DEC) in stool samples of children (under 5 years) with acute diarrhea.  Methods: Samples were collected from January 2011 to December 2011, from Dhaka Medical College Hospital and Dhaka Shishu Hospital. Multiplex PCR with five specific primer pairs to detect enteropathogenic E. coli (eae, bfp), enterotoxigenic E. coli (lt, st) and enteroaggregative E. coli (aat) were used. However, enteroinvasive E. coli, enterohemorrhagicE. coli and diffusely adhererentE. coli were not sought. Result: In total, 135 (67.5%) E. coli were isolated from 200 stool samples. The prevalence of DEC was 68 (34%). Among DEC, most frequently isolated pathotype was EPEC 40 (58.82%), followed by ETEC 24 (35.29%) and EAggEC 18 (26.47%). Among the EPEC, 5 (12.5%) were typical EPEC. Among the 68 DEC positive cases, 22 samples contained more than one pathogenic gene in various combinations. Among the combination of DEC, EPEC+ETEC combination was 6 (27.27%) followed by ETEC+EAggEC 4 (18.18%), EPEC+EAggEC and ETEC+EPEC+EAggEC were both in 3 (13.6%). Conclusion:This study shows that DEC is a common cause of childhood diarrhea in Dhaka city of Bangladesh. By using multiplex PCR assay, DEC can be diagnosed in one PCR reaction that makes a conclusive diagnosis of diarrhea. DOI: http://dx.doi.org/10.3126/ajms.v5i2.8576 Asian Journal of Medical Science, Volume-5(2) 2014: 59-66

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

2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
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.

Microbial Profile and Antibiotic Susceptibility of Campylobacter spp. and Salmonella spp. in Broilers Processed in Air-Chilled and Immersion-Chilled Environments

2002 ◽  
Vol 65 (6) ◽  
pp. 948-956 ◽  
Author(s):  
MARCOS X. SÁNCHEZ ◽  
WADE M. FLUCKEY ◽  
MINDY M. BRASHEARS ◽  
SHELLY R. McKEE
Keyword(s):  
Antibiotic Resistance ◽  
Bacterial Growth ◽  
Cross Contamination ◽  
Salmonella Spp ◽  
E Coli ◽  
Microbial Profile ◽  
Resistance Patterns ◽  
Poultry Processing ◽  
Antibiotic Resistance Patterns ◽  
Campylobacter Spp

Carcass chilling is considered a critical step for inhibiting bacterial growth during poultry processing. The objective of this study was to compare microbiological loads and the incidence of Salmonella spp. and Campylobacter spp. on broiler carcasses subjected to immersion chilling and air chilling. Additionally, the antibiotic resistance patterns of pathogen isolates were determined. The results of this study indicated that the incidence of Salmonella spp. and Campylobacter spp. tends to be significantly lower in air-chilled broilers, suggesting that cross-contamination may be more prevalent for immersion-chilled broilers. No significant differences were detected between chilling treatments for total aerobic populations or for generic E. coli or coliform counts. Psychrotrophic populations were significantly larger (P &lt; 0.05) in immersion-chilled broilers than in their air-chilled counterparts. Campylobacter isolates from immersion-chilled broilers had a higher incidence of resistance to nalidixic acid (NAL) and related fluoroquinolones than isolates from air-chilled broilers did. Additionally, Campylobacter isolates from air-chilled broilers had a higher frequency of resistance to tetracycline than isolates from immersion-chilled broilers did. With regard to Salmonella, isolates from immersion-chilled broilers had a higher incidence of resistance to NAL than isolates from air-chilled samples did. No Salmonella isolates from immersion- or air-chilled broilers were resistant to the fluoroquinolonestested. The chilling method used during processing may influence the microbial profile of postchilled broilers.

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