Identification, characterization and molecular epidemiology of Escherichia coli isolated from lamb and goat kids with diarrhoea (original) (raw)
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Zoonoses and Public Health, 2009
The purpose of this study was to determine aetiological agents of diarrhoea in neonatal calves and to investigate virulence gene markers of Escherichia coli strains isolated from calves by multiplex polymerase chain reaction (PCR). Eighty-two diarrhoeic calves and 18 healthy calves were used as subjects. Faeces were taken from the rectums of all the calves and were subjected to bacterial culture. Antigen enzyme-linked immunosorbent assay (ELISA) was performed to detect rotavirus, coronavirus and E. coli K99 in faeces of all the calves. A multiplex PCR was used to characterize E. coli strains in all the calves. Escherichia coli was isolated from 37 faeces samples, Enterococcus ssp. was isolated from 22 faeces samples and Salmonella was isolated from one faeces sample in diarrhoeic calves. Furthermore, only E. coli was isolated from all 18 faeces samples of healthy calves. Of the 37 E. coli isolated from diarrhoeic calves, K99 (18.9%), F41 (18.9%), heat-stable enterotoxin a (STa) (18.9%), Shiga toxin 1 (Stx1; 13.5%) and Shiga toxin 2 (Stx2; 5.4%) and intimin (8.1%) genes were identified by multiplex PCR. Of the 18 E. coli isolated from healthy calves, K99 (16.6%) and intimin (55.5%) genes were identified by PCR. A total of 15 rotavirus, 11 coronavirus and 11 E. coli K99 were detected in diarrhoeic calves by the antigen ELISA. As a result, this study shows that rotavirus, coronavirus, E. coli and Enterococcus ssp. were determined to play a role in the aetiology of diarrhoea in the neonatal calves. K99, F41, STa, Stx1 and Stx2 were found as the most common virulence gene markers of E. coli strains isolated from calves with diarrhoea. Multiplex PCR may be useful for characterization of E. coli isolated from calves.
Open Vet J. 9:65-73, 2019
Enterotoxigenic Escherichia coli (ETEC) is the most common and global cause of neonatal calf diarrhea, but there is a little information regarding calf ETEC strains in Argentina. In this study, five ETEC isolates from diarrheic dairy calves (2-10 d old) from Buenos Aires and Cordoba, Argentina were characterized on the basis of virulence gene (VG) pattern, O:H serotyping, hemolytic phenotype, phylogenetic group affiliation, antimicrobial (AM) resistance profile, and presence of integron class 1 and 2. The five isolates were examined by polymerase chain reaction (PCR) for the presence of 18 bovine VGs and showed the following genotypes: F5 + /F41 + /sta + (D242), F5 + /sta + (D158), F5 + /sta + (D157), F5 + (D151-9), and F5 + /iucD + (D151-5). These VGs confer pathogenic potential and most of them are associated with the ETEC pathotype. The five isolates showed a non-hemolytic phenotype, belonged to five different serotypes: O101:H − , O141:H − , O60:H − , ONT:H10, and ONT:H − , and were assigned to the phylogenetic group A by the quadruplex Clermont PCR method. The AM resistance of the three isolates D242, D157, and D151-5 was determined by agar disk diffusion method for 24 AMs and they exhibited a multi-resistance phenotype (resistance to four different AM classes: Cephalosporins, Penicillins, Macrolides, and Ansamycins). In addition, class 1 integrons were found in the isolate D151-5 containing the dfrA17-aadA5 gene cassette and in the bovine ETEC reference strain FV10191 containing the dfrA1-aadA1 gene cassette. The present study revealed for the first time the occurrence of multi-resistant ETEC associated with neonatal diarrhea in dairy calves in Argentina. This finding may be used for diagnostic and therapeutic purposes.
Semina: Ciências Agrárias, 2021
Neonatal diarrhea is the main cause of early mortality and morbidity in farm animals and the source of huge, direct and indirect, economic husbandry losses. Escherichia coli, a common harmless commensal bacterium, can turn into a main diarrheal pathogen through antibiotic resistance and the expression of genetically acquired virulence factors. In this study, fecal samples obtained from eight farms of animals with clinical signs characteristic of diarrhea were subjected to culture and bacterial isolation. Colonies suggestive of E. coli were identified through morphological and biochemical characteristics. Susceptibility tests to the main veterinary antibacterial agents were conducted using agar disk diffusion followed by phenotypical detection of extended-spectrum ?-lactamase (ESBL). A total of 301 colonies were characterized as E. coli and, out of the 192 that were tested, 134 showed resistance to three or more classes of antimicrobial drugs and were classified as multidrug resistan...
Advances in Animal and Veterinary Sciences, 2019
H ealth of the newborn calves is a matter of balance concerning the pathogens, immune system of the newborn animal, environment and management system. Infection in newborn calves is multifactorial in nature and it seems to be the result of interaction between different factors that may contribute to the buildup of infection including the immunological, nutritional, and environmental factors, (Hosein, 2018). Neonatal calf diarrhea is one of the most serious health problems in the livestock industry and an important cause of economic losses due to high morbidity and mortality rates, high treatment costs and low growth rate, (Al-Alo et al., 2018; Anderson et al., 2003; Bazeley, 2003). Diarrhea is the most important cause of losses in calves up to 30 days of age and still problematic, likely because of the multi-factorial nature of the disease, Gomez and Weese (2017). Difference between health and disease among new born calves is very often just a slight tip of a delicate balance that weighs calf and environmental factors with the pathogens to which the calf will be exposed. E. coli an important enteric pathogen of bovine neonates, is established in intestines shortly after the birth and remains throughout life. It has been incriminated as a major cause of diarrhea characterized by progressive dehydration and death that may occur depending on the age of the calf when scour started and the pathotypes of E. coli involved (Nguyen et al., 2011). Several E. coli serotypes, causing morbidity and mortality, have been isolated from calves suffering from diarrhea (Wani et al., 2003). E. coli pathotypes that are incriminated in neonatal calf diar-research Article Abstract | A total of 14 (30.43%) diarrheic calves of 1-4 weeks out of 46 calves kept under poor hygienic conditions in a dairy farm were suffering clinically from acute diarrhea. Diarrheic calves showed fever, diarrhea, dehydration, pneumonia recumbence and death of two calves. Thirteen (92.85 %) isolates of E. coli out of the 14 collected rectal swaps of diarrheic calves were identified on bacteriological and molecular basis. The results of the amplification of phoAgene using PCR revealed that all E. coli isolates showed positive result for the presence of phoAgene, thus confirming their identity as E. coli. All tested E. coli isolates were positive for intimin (eae) A, attaching and effacing gene (gene species specific) for E. coli (100%). No isolate had shiga toxin1 (stx1), shiga toxin 2 (stx2), hemolysin (hylA), and E. coli enterotoxin genes heat stable enterotoxin (st) and heat labile enterotoxin (lt). The obtained results indicate the possible participation of pathogenic E. coli in calf diarrhea. EPEC represented 100% of the tested E. coli strains obtained from diarrheic calves. The obtained results indicated that EPEC infection is a major health problem among calves and suggests the significance of poor hygiene measures in the investigated farm the possible participation of calves in the zoonotic transmission of pathogenic E. coli.
Open veterinary journal, 2019
Enterotoxigenic Escherichia coli (ETEC) is the most common and global cause of neonatal calf diarrhea, but there is a little information regarding calf ETEC strains in Argentina. In this study, five ETEC isolates from diarrheic dairy calves (2-10 d old) from Buenos Aires and Cordoba, Argentina were characterized on the basis of virulence gene (VG) pattern, O:H serotyping, hemolytic phenotype, phylogenetic group affiliation, antimicrobial (AM) resistance profile, and presence of integron class 1 and 2. The five isolates were examined by polymerase chain reaction (PCR) for the presence of 18 bovine VGs and showed the following genotypes: F5 + /F41 + /sta + (D242), F5 + /sta + (D158), F5 + /sta + (D157), F5 + (D151-9), and F5 + /iucD + (D151-5). These VGs confer pathogenic potential and most of them are associated with the ETEC pathotype. The five isolates showed a non-hemolytic phenotype, belonged to five different serotypes: O101:H − , O141:H − , O60:H − , ONT:H10, and ONT:H − , and were assigned to the phylogenetic group A by the quadruplex Clermont PCR method. The AM resistance of the three isolates D242, D157, and D151-5 was determined by agar disk diffusion method for 24 AMs and they exhibited a multi-resistance phenotype (resistance to four different AM classes: Cephalosporins, Penicillins, Macrolides, and Ansamycins). In addition, class 1 integrons were found in the isolate D151-5 containing the dfrA17-aadA5 gene cassette and in the bovine ETEC reference strain FV10191 containing the dfrA1-aadA1 gene cassette. The present study revealed for the first time the occurrence of multi-resistant ETEC associated with neonatal diarrhea in dairy calves in Argentina. This finding may be used for diagnostic and therapeutic purposes.
This study was performed to estimate the prevalence of Enterotoxigenic Escherichia coli (ETEC) in calves' diarrhoea samples by molecular and serological methods. Rectal swabs from 127 diarrhoeaic calves were collected from 10 different farms of Tehran province during March to September 2014. ETEC was detected by both mPCR and ELISA. According to the results, ETEC was detected in 14 (11%) and 5 (3.9%) diarrhoea samples by mPCR and ELISA methods respectively. All 14 positive samples were detected by mPCR method had three virulence genes including K99 (F5), F41 and STa. Although 11 (78.5%) and 3 (21.4%) positive cases were identified from traditional and semi-industrial farms respectively, no isolates were identified related to the industrial farms. The results represented that detection assays such as ELISA which is only able to detect fimbriae (F5), will follow false-positive cases. For this reason mPCR assay has been developed and recognized as effective, rapid and reliable tools for detection of ETEC.
Tropical Animal Health and Production
Escherichia coli field isolates from calves were characterized and categorized into the most significant diarrheagenic pathotypes using polymerase chain reaction (PCR) assays with different specific primers. The used PCR systems were designed to detect sequences representing the group-specific virulence genes encoding fimbriae f5 (K99), Shiga toxins (stx 1 and stx 2), heat-stable enterotoxins (st), heat-labile enterotoxins (lt), intimin (eae), hemolysin (hylA), and EAEC heat-stable enterotoxin (astA). In the present work, a total of 150 E. coli field isolates were recovered from 150 fecal swabs collected from 100 diarrheic and 50 apparently healthy in-contact cattle and buffalo calves under 3 months old. Out of these 150 isolated E. coli, 106 isolates from 77 diarrheic and 29 in-contact calves harbored one or more of the investigated virulence genes. The pathotyping of the isolates could classify them into shigatoxigenic E. coli (STEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), and enteroaggregative E.
Epidemiological study of diarrheagenic Escherichia coli virulence genes in newborn calves
Pakistan Veterinary Journal, 2016
Received: Revised: Accepted: Online available: June 20, 2015 September 14, 2015 September 27, 2015 January 09, 2016 Diarrheagenic Escherichia coli (DEC) are considered one of the major etiological agents of neonatal calf diarrhea (NCD). One hundred calves (2-21 days-old) suffering from diarrhea in Sharkia Governorate, Egypt were used in this study. Six E. coli virulence genes were characterized by multiplex PCR including; heat-stable enterotoxins (STa; 33.3%), heat-labile enterotoxin (LT; 30%), two Shiga toxins: (Stx1; 86.67%), (Stx2; 26.67%) produced from Shiga toxin producing E. coli (STEC) and two enterotoxigenic E. coli (ETEC) adhesions (F5; 13.3%) and (F41; 16.67%). Each isolate was found to carry one or more virulence genes. Clinicobiochemical examination showed significant (P<0.01) increase in serum total protein, globulin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine, blood urea nitrogen (BUN), and potassium levels. Meanwhile; serum albumin...
Tropical Animal Health and Production, 2017
The aim of the present study was to determine the analysis of virulence genes and antimicrobial profile of diarrheagenic Escherichia coli isolated from diseased lambs. Two hundred ninety E. coli isolates were recovered from 300 rectal swabs of diarrheic lambs and were confirmed by biochemical tests. The pathotype determination was done according to the presence of genes including f5, f41, LTI, STI, bfp, ipaH, stx 1 , stx 2 , eae, ehlyA, cnf 1 , cnf 2 , cdIII, cdIV, and f17 by PCR method. Sixty-six isolates (23.72%) possessed the STI gene and categorized into entrotoxigenic E. coli (ETEC). Nine isolates (3.1%) and five isolates (1.72%) were positive for the cnf1 and cnf2 genes which categorized into necrotoxic E. coli (NTEC). Hundred and seventeen isolates (40.34%) harbored stx 1 and/or stx 2 and classified as Shiga toxin-producing E. coli (STEC). Thirteen isolates (4.48%) were assigned to atypical entropathogenic E. coli (aEPEC) and possessed eae gene. Two isolates (0.68%) were positive for ipaH gene and were assigned to entroinvasive E. coli (EIEC). Statistical analysis showed a specific association between eae gene and STEC pathotype (P < 0.0001). The most prevalent resistance was observed against lincomycin (96.5%) and the lowest resistance was against kanamycine (56.02%), respectively. The high prevalence of STEC and ETEC indicates that diarrheic lambs represent an important reservoir for humans. ETEC may play an important role for frequent occurrence of diarrhea in lambs observed in this region. Due to high antibiotic resistance, appropriate control should be implemented in veterinary medicine to curb the development of novel resistant isolates.