Molecular identification of diarrheagenic Escherichia coli pathotypes and their antibiotic resistance patterns among diarrheic children and in contact calves in Bahir Dar city, Northwest Ethiopia
Abstract:Diarrheagenic Escherichia coli strains are an essential cause of diarrheal infection in younger children and animals. The study was focused on understanding the associated characteristics of various DEC strains among children and calves, establishing the possible zoonotic transmission, and determining their antibiotic resistance patterns. Samples from 144 acute diarrheic children and 50 diarrheic calves were collected and processed using traditional culture methods. The molecular identification of pathotypes w… Show more
“…In the present study, E. coli isolates were found sensitive to ciprofloxacin (94.7%), kanamycin (89.5%), and chloramphenicol (84.2%). This sensitivity result agrees with the findings of Belete et al [ 24 ] and Dejene et al [ 15 ]. However, isolates were found resistant to amoxicillin (89.5%), in line with 64.9% resistance in a study conducted by Belete et al [ 24 ] in Bahir Dar town.…”
Section: Discussionsupporting
confidence: 93%
“…Dairy production is practiced in and around the town, supporting the livelihood of the inhabitants of Bahir Dar. The mean annual rainfall of the city is about 596 mm, with a bimodal rainy season distribution of 70%, which occurs from June to September and 30% from March to April [ 24 ]. Fig.…”
Section: Methodsmentioning
confidence: 99%
“…The PCR assays were performed using four specific primers ( Table 1 ), targeting the stx1 , stx2 , eae , and ribo157 genes as previously described [ 24 , 28 ]. The DNA was extracted using a Qiagen DNeasy Blood and Tissue Kit based on the manufacturer's instructions (Qiagen, Germantown, MD, USA).…”
Section: Methodsmentioning
confidence: 99%
“…Mueller Hinton (MHA) agar media (Oxoid, England) and a 0.5 Mac-Farland standard suspension of the bacteria were prepared to culture and adjust the density of the bacterial suspension, respectively. The result of the zone of inhibition was measured and interpreted as sensitive, intermediate, or resistant according to the Clinical and Laboratory Standards Institute (CLSI) guidelines [ 24 ].…”
“…In the present study, E. coli isolates were found sensitive to ciprofloxacin (94.7%), kanamycin (89.5%), and chloramphenicol (84.2%). This sensitivity result agrees with the findings of Belete et al [ 24 ] and Dejene et al [ 15 ]. However, isolates were found resistant to amoxicillin (89.5%), in line with 64.9% resistance in a study conducted by Belete et al [ 24 ] in Bahir Dar town.…”
Section: Discussionsupporting
confidence: 93%
“…Dairy production is practiced in and around the town, supporting the livelihood of the inhabitants of Bahir Dar. The mean annual rainfall of the city is about 596 mm, with a bimodal rainy season distribution of 70%, which occurs from June to September and 30% from March to April [ 24 ]. Fig.…”
Section: Methodsmentioning
confidence: 99%
“…The PCR assays were performed using four specific primers ( Table 1 ), targeting the stx1 , stx2 , eae , and ribo157 genes as previously described [ 24 , 28 ]. The DNA was extracted using a Qiagen DNeasy Blood and Tissue Kit based on the manufacturer's instructions (Qiagen, Germantown, MD, USA).…”
Section: Methodsmentioning
confidence: 99%
“…Mueller Hinton (MHA) agar media (Oxoid, England) and a 0.5 Mac-Farland standard suspension of the bacteria were prepared to culture and adjust the density of the bacterial suspension, respectively. The result of the zone of inhibition was measured and interpreted as sensitive, intermediate, or resistant according to the Clinical and Laboratory Standards Institute (CLSI) guidelines [ 24 ].…”
“…AMR among pathogenic E. coli strains are steadily increasing in Ethiopia [7,21]. Despite the growing threat of antimicrobial resistance in our country, little attention is given to its management, and novel alternatives have yet to be explored.…”
Escherichia coli is a common fecal coliform, facultative aerobic, gram-negative bacterium. Pathogenic strains of such microbes have evolved to cause diarrhea, urinary tract infections, and septicemias. The emergence of antibiotic resistance urged the identification of an alternative strategy. The use of lytic bacteriophages against the control of pathogenic E. coli in clinics and different environmental setups (waste and drink water management) has become an alternative therapy to antibiotic therapy. Thus, this study aimed to isolate and characterize lytic bacteriophage from various sources in Addis Ababa, tested them against antimicrobial-resistant diarrheagenic E. coli strains and evaluated their therapeutic potential under in vitro conditions. A total of 14 samples were processed against six different diarrheagenic E. coli strains. The conventional culture and plaque analysis agar overlay method was used to recover lytic bacteriophage isolates. The phage isolates were characterized to determine their lytic effect, growth characteristics, host range activity and stability under different temperature and pH conditions. Phage isolates were identified by scanning electron microscope (SEM), and molecular techniques (PCR). In total, 17 phages were recovered from 84 tested plates. Of the 17 phage isolates, 11 (65%) were Myoviridae-like phages, and 6 (35%) phage isolates were Podoviridae and Siphoviridae by morphology and PCR identification. Based on the host range test, growth characteristics and stability test 7 potent phages were selected. These phages demonstrated better growth characteristics, including short latent periods, highest burst sizes, and wider host ranges, as well as thermal stability and the ability to survive in a wide range of pH levels. The promising effect of these phages against AMR pathogens has raised the possibility of their use in the biological control of bacterial infections.
Enterotoxigenic Escherichia coli (ETEC) is one of the leading causes of infant diarrhea in low- and middle-income countries (LMICs). Diarrheal pathogens are transmitted through environmental reservoirs; however, the bacterial clones that spread across the human-environment interphases remind unexplored. We aimed to determine the relationship and clonal dissemination of ETEC between children with diarrhea (> 5 years of age) and polluted water samples from local river in La Paz, Bolivia. Our study used whole genome sequencing and phenotypic fingerprinting system (PhenePlates) to analyze ETEC strains. We showed that ST218 and ST410 LT+STh CS23 ETEC were found with high frequency in both samples. The CS23 ETEC isolates were found within several STs, E. coli phylogroups A, B1, C, and D, and across ETEC lineages. Our comparative genomic analysis and PhenePlate screening of globally distributed clinical ETEC strains suggested that virulent CS23 plasmids acquisition occurs independently of the bacterial chromosomal background. Environmental strains were more often multidrug-resistant (MDR) than clinical isolates and harbored the class 1 integron-integrase gene intI1 next to the MDR cassettes. Retrospective analysis of antibiotic resistance in ETEC revealed a high frequency of MDR in clinical isolates. The LT+STh CS23 ETEC isolates showed an increased biofilm ability at environmental temperature, equal cytotoxicity, and significantly lower adherence to human epithelial cells compared to ETEC expressing other CFs. Together, our findings suggest that CS23 is more prevalent in ETEC than previously estimated, and the Choqueyapu River is a reservoir for LT+STh CS23 ETEC containing strains capable of causing diarrheal cases in children.
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