“…Bacterial genomic DNA extraction using the boiling method and the molecular confirmation of the presumptive E. coli isolates were done as described by Iwu et al [ 19 ]. Verification of PCR amplification products was performed in 1.5% agarose gel stained with ethidium bromide and electrophoresed at 120 V for 45 min using 0.5 × Tris-borate-EDTA (TBE) buffer and then viewed in UV transilluminator (ALLIANCE 4.7).…”
Background. Diarrhea has been reported as the leading cause of childhood mortality and morbidity globally but with disproportionate impacts in developing nations. Among bacterial etiologic agents of diarrhea, diarrheagenic Escherichia coli is the main cause of the disease among children under the age of 5 years. This study is aimed at determining the prevalence and antibiogram pattern of diarrheagenic Escherichia coli (DEC) pathotypes associated with diarrhea cases in the study area. Methods. A total of 120 presumptive isolates of E. coli were obtained from diarrheal stool samples from male and female patients below 12 years of age using chromogenic agar. Confirmation of the isolates and screening for virulence genes were determined by polymerase chain reaction (PCR) while antimicrobial susceptibility testing was performed using the disk diffusion method. The presence of antibiotic resistance genes to chloramphenicol and tetracycline among the confirmed isolates was also profiled by PCR based on the observed phenotypic resistance pattern. Results. Of the 120 presumptive isolates, 88.3% (106/120) were confirmed as E. coli through PCR. The molecular pathotyping of the confirmed isolates showed their distribution as 41% (43/106) of diffusely adhering E. coli (DAEC), 17% (18/106) of enterohemorrhagic E. coli (EHEC), 17% (18/106) of enteropathogenic E. coli (EPEC), and 10% (11/106) of enteroinvasive E. coli (EIEC), while enteroaggregative E. coli (EAEC) and enterotoxigenic E. coli (ETEC) were not detected, and the remaining 15% did not belong to any pathotype. Notably, high resistance of the isolates to commonly used antimicrobials was observed as follows: ampicillin (98%), chloramphenicol (94%), trimethoprim-sulfamethoxazole (96%), and tetracycline (90.6%), while a relatively low number of the confirmed isolates were resistant to ciprofloxacin (45%) and imipenem (36%). In addition, 94% of the isolates that exhibited phenotypic resistance against chloramphenicol harbored the catA1 resistance gene while 89% that showed resistance to tetracycline had tetA genes. Conclusions. These findings showed that DEC could be considered as the leading etiologic bacterial agent responsible for diarrhea in the study community, and the observable high degree of resistance of the isolates to antimicrobial agents is of huge significance, calling for stakeholders to adopt and consolidate the existing antimicrobial stewardship scheme of the government, in order to ensure an uncompromised public health.
“…Bacterial genomic DNA extraction using the boiling method and the molecular confirmation of the presumptive E. coli isolates were done as described by Iwu et al [ 19 ]. Verification of PCR amplification products was performed in 1.5% agarose gel stained with ethidium bromide and electrophoresed at 120 V for 45 min using 0.5 × Tris-borate-EDTA (TBE) buffer and then viewed in UV transilluminator (ALLIANCE 4.7).…”
Background. Diarrhea has been reported as the leading cause of childhood mortality and morbidity globally but with disproportionate impacts in developing nations. Among bacterial etiologic agents of diarrhea, diarrheagenic Escherichia coli is the main cause of the disease among children under the age of 5 years. This study is aimed at determining the prevalence and antibiogram pattern of diarrheagenic Escherichia coli (DEC) pathotypes associated with diarrhea cases in the study area. Methods. A total of 120 presumptive isolates of E. coli were obtained from diarrheal stool samples from male and female patients below 12 years of age using chromogenic agar. Confirmation of the isolates and screening for virulence genes were determined by polymerase chain reaction (PCR) while antimicrobial susceptibility testing was performed using the disk diffusion method. The presence of antibiotic resistance genes to chloramphenicol and tetracycline among the confirmed isolates was also profiled by PCR based on the observed phenotypic resistance pattern. Results. Of the 120 presumptive isolates, 88.3% (106/120) were confirmed as E. coli through PCR. The molecular pathotyping of the confirmed isolates showed their distribution as 41% (43/106) of diffusely adhering E. coli (DAEC), 17% (18/106) of enterohemorrhagic E. coli (EHEC), 17% (18/106) of enteropathogenic E. coli (EPEC), and 10% (11/106) of enteroinvasive E. coli (EIEC), while enteroaggregative E. coli (EAEC) and enterotoxigenic E. coli (ETEC) were not detected, and the remaining 15% did not belong to any pathotype. Notably, high resistance of the isolates to commonly used antimicrobials was observed as follows: ampicillin (98%), chloramphenicol (94%), trimethoprim-sulfamethoxazole (96%), and tetracycline (90.6%), while a relatively low number of the confirmed isolates were resistant to ciprofloxacin (45%) and imipenem (36%). In addition, 94% of the isolates that exhibited phenotypic resistance against chloramphenicol harbored the catA1 resistance gene while 89% that showed resistance to tetracycline had tetA genes. Conclusions. These findings showed that DEC could be considered as the leading etiologic bacterial agent responsible for diarrhea in the study community, and the observable high degree of resistance of the isolates to antimicrobial agents is of huge significance, calling for stakeholders to adopt and consolidate the existing antimicrobial stewardship scheme of the government, in order to ensure an uncompromised public health.
“…Molecular confirmation of presumptive S. aureus and E. coli isolates was done by PCR using a primer pair to target the thermonuclease (Nuc) gene for S. aureus [ 14 , 38 ] and uidA gene for E. coli [ 39 , 40 ] (Figures 1 and 2 ). Quality control strains S. aureus ATCC 25923 and E. coli ATCC 25922 served as positive controls.…”
Section: Methodsmentioning
confidence: 99%
“…Multiple antibiotic resistance phenotypes (MARPs) for S . aureus isolates from the formal and informal meat sectors were then generated for isolates that were resistant to five or more antimicrobials [ 39 ]. The frequencies, percentages, and number of antimicrobials to which the isolates were resistant and resistance patterns were obtained from the antimicrobial susceptibility testing (AST).…”
Background. Foodborne diseases (FBD) caused by resistant pathogens are a global public health problem. One main driver of the increasing FBD incidence is the transfer of pathogenic organisms from animal guts to carcasses during processing and subsequent transfer from meat products to consumers. Methods. In this study, meat samples from abattoirs in the formal meat sector (FMS) (n=140) and slaughter points in the informal meat sector (IMS) (n=104) were collected for microbial detection and phenotypic AMR determination using polymerase chain reaction. Results. The antibiogram of Staphylococcus aureus isolates revealed that resistance to clindamycin (74.3%) and ampicillin (59.5%) was highest in the FMS, while resistance to penicillin (83.8%) and tetracycline (82.1%) was highest in the IMS. Escherichia coli isolates show significant resistance to chloramphenicol (90.7%) and tetracycline (82.3%) in the FMS. Likewise, resistance to tetracycline (92.3%) and sulfamethoxazole/trimethoprim (87.5%) was highest in the IMS. The multiple antibiotic resistance index (MARI) for S. aureus and E. coli ranged from 0.3 to 0.8 and 0.2 to 0.5, respectively. Conclusion. This study suggests high-level contamination of meat with resistant pathogens and highlights the public health consequences associated with consuming such unhygienic products.
“…Escherichia coli is a ubiquitous gut microorganism which forms part of the natural flora of the gastrointestinal system. Its ability to acquire both resistant determinants and virulence factors has been acknowledged by many researchers [3].…”
This study aimed to characterise antibiotics resistance of Escherichia coli isolates from the formal meat sector (FMS) and informal meat sectors (INMS).
MethodA total of 162 and 102 E. coli isolates from the FMS, and INMS respectively were isolated by standard culture-based, and biochemical reactions. The isolates were further confirmed by polymerase chain reaction (PCR). The disc diffusion method was used to screen for antimicrobial susceptibility against 19 different antibiotics. The presence of class 1-2 integrons in each E. coli isolates was assessed using 3 0 -CS and 5 0 -CS regions specific primers.
ResultAmong the 19 antimicrobials, resistance to tetracyclines, aminoglycosides, cephalosporins, and nitrofurans were found to be more frequent than carbapenems and chloramphenicol. The number of multi-drug resistance ranged from three to ten antimicrobials. The resistant determinants with the highest prevalence in the FMS and INMS were;
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