Investigation of plasmid-mediated resistance in E. coli isolated from healthy and diarrheic sheep and goats.

Escherichia coli is zoonotic bacteria and the emergence of antimicrobial-resistant strains becomes a critical issue in both human and animal health globally. This study was therefore aimed to investigate the plasmid-mediated resistance in E. coli strains isolated from healthy and diarrheic sheep and goats. A total of 234 fecal samples were obtained from 157 sheep (99 healthy and 58 diarrheic) and 77 goats (32 healthy and 45 diarrheic) for the isolation and identification of E. coli. Plasmid DNA was extracted using the alkaline lysis method. Phenotypic antibiotic susceptibility profiles were determined against the three classes of antimicrobials, which resistance is mediated by plasmids (Cephalosporins, Fluoroquinolone, and Aminoglycosides) using the disc-diffusion method. The frequency of plasmid-mediated resistance genes was investigated by PCR. A total of 159 E. coli strains harbored plasmids. The isolates antibiogram showed different patterns of resistance in both healthy and diarrheic animals. A total of (82; 51.5%) E. coli strains were multidrug-resistant. rmtB gene was detected in all Aminoglycoside-resistant E. coli, and the ESBL-producing E. coli possessed different CTX-M genes. Similarly, fluoroquinolone-resistant E. coli possessed different qnr genes. On the analysis of the gyrB gene sequence of fluoroquinolone-resistant E. coli, multiple point mutations were revealed. In conclusion, a high prevalence of E. coli with high resistance patterns to antimicrobials was revealed in the current study, in addition to a wide distribution of their resistance determinants. These findings highlight the importance of sheep and goats as reservoirs for the dissemination of MDR E. coli and resistance gene horizontal transfer.

Diagnostic tools of caprine and ovine anaplasmosis: a direct comparative study.

BACKGROUND: The diagnosis of anaplasmosis is rather conflicting with other haemoprotozoans. Hence, the study aimed to compare and evaluate the efficiency of competitive ELISA (cELISA), indirect fluorescence antibody (IFA), and Polymerase chain reaction (PCR) for precise diagnosis of Anaplasma spp. and to assess their concordance with microscopic examination (ME). RESULTS: A total of 312 blood samples (189 sheep and 123 goats) were examined for Anaplasma infection during a 1 year period. Giemsa-stained blood smears were examined under the microscope. IFA and cELISA were used for the detection of Anaplasma spp. antibodies. PCR was used as a standard of truth and for the identification of Anaplasma species. Using cELISA assay, 47.4% (148) were positive (93 sheep and 55 goats) with a sensitivity and specificity of 91.9, and 86.9%, respectively. Using IFA, it was found that 57.4% (179)were positive (113 sheep and 66 goats) with a sensitivity and specificity of 100, and 93.3%, respectively. PCR assay identified A. ovis in 49 (25.3%) sheep and 30 (15.5%) goats, and A. phagocytophilumin 74 (38.1%) sheep and 41 (20.8%) goats. CONCLUSIONS: High sensitivity and specificity values of IFA and ELISA tests compared to microscopic examination strongly support their utility in the diagnosis of Anaplasma infection. PCR was a more specific diagnostic tool that allows to discriminate between Anaplasma subspecies, which makes it the method of choice for anaplasmosis diagnosis.

Investigation of viral and bacterial enteropathogens of diarrheic sheep and goats in Medina, Saudi Arabia.

Diarrhea is a serious problem in sheep and goat farming, causing great economic losses. Most viral and bacterial enteropathogens of diarrheic sheep and goats are considered foodborne pathogens with a potential to be zoonotic. The present study investigated the prevalence of some viral and bacterial enteropathogens in diarrheic sheep and goats between October 2015and February 2016 in Medina, Saudi Arabia. A total of 310 fecal samples were collected from diarrheic sheep (n=193) and goats (n=117). The samples were screened for the presence of rotavirus and bovine coronavirus (BCoV) using Sandwich (ELISA) technique. The bacterial enteropathogens were isolated and identified biochemically and the virulence factors of Escherichia coli were determined by using PCR. Escherichia coli was the most prevalent agent in both sheep and goats (34.7% and 30.7%, respectively). According to the expressed virulence genes, Enterotoxigenic Escherichia coli (ETEC) was detect in 34.3% of sheep isolates and 30.6% of goats isolates. Salmonella species was isolated from 3.6% of sheep and 2.6% of goats. Klebsiella species was isolated from 1.6% of sheep but not from goats. Regarding viral agents, rotavirus was found in 31.6% of sheep and 27.4% of goats, while BCoV was detected in 19.6% of the sheep and 16.2% of the goats. The prevalence of bacterial and viral enteropathogens was significantly higher in the 0-12 months age group compared to the older age groups. Double infection was the most common (53.0%) infection pattern compared to single (37.5%) and triple (9.5%) infections. Rotavirus infection was significantly associated with ETEC infection. In conclusion, we report high prevalence of rotavirus and ETEC in sheep and goats, which are of veterinary and public health importance. This study provides valuable data on the prevalence of viral and bacterial enteropathogens in sheep and goats in Medina, Saudi Arabia that will be useful to develop control measures for these pathogens.

Molecular studies on diarrhea-associated Escherichia coli isolated from humans and animals in Egypt.

UNLABELLED: The molecular characteristics of Escherichia coli isolates from Egypt and the relationship of E. coli strains from claves, camels and humans are limited. We analyzed the genetic relationships of 48 diarrhea-associated E. coli strains isolated from sporadic diarrheal cases from humans (n=26), calves (n=14) and camels (n=8) using multilocus sequence type (MLST), virulence genes, and pulsed field gel electrophoresis (PFGE). Enterotoxigenic E. coli (ETEC) accounted for 60.4% of all samples and the rest were Enteropathogenic E. coli (EPEC) 10.4%, Diffuse adhering E. coli (DAEC) 8.3%, Enteroaggreagative E. coli (EAEC) 6.3%, Verotoxigenic E. coli (VTEC) 6.3%, Untypable E. coli. 6.3% and Atypical enteropathogenic E. coli (aEPEC) 2.1%. We identified 17 new sequence types (ST) and 12 new alleles. Generally, strains divided into 6 clonal complexes, and clonal complex (CC) 10 was the major one, detected in (15/48; 31.3%) strains from humans, calves and camels. The close relationship among the strains from different hosts was regarding to mdh, purA, and recA genes which presented a minor variation in relation to other housekeeping genes. CONCLUSION: MLST analysis suggested an endemic prevalence of clonal complex (CC) 10 in Egypt. Same sequencing types (ST) could be detected in human, calf and camel, especially ST10, indicating the ability of E. coli to cross the host barrier. Together with PFGE results and virulence genotypes we conclude that human, calf and camel can be colonized and infected with similar E. coli strains and provide evidence of calves and camels role as a reservoir for similar strains of diarrhea-associated E. coli.

Molecular characterization and susceptibility of methicillin-resistant and methicillin-susceptible Staphylococcus aureus isolates from hospitals and the community in Vladivostok, Russia.

A prospective study was conducted during an 8-month period, from August 2006 to April 2007, to describe the epidemiology of Staphylococcus aureus-associated infections. In addition, the molecular characteristics, antimicrobial susceptibilities and antibiotic resistance determinants were identified in S. aureus isolates from hospitals and the community in Vladivostok, Russia. Among the 63 S. aureus isolates eligible for this study, methicillin resistance was observed in 48% (n = 30). Hospital-acquired strains accounted for 93% (28/30) of all methicillin-resistant S. aureus (MRSA) isolates. The major MRSA clone (sequence type (ST) 239, staphylococcal cassette chromosome mec (SCCmec) type III, Panton-Valentine leukocidin (PVL)-negative, with two related staphylococcal protein A gene (spa) types (types 3 and 351)) represented 90% of all of the MRSA isolates. This clone was multidrug-resistant, and 41% of isolates showed resistance to rifampicin. Community-acquired MRSA isolates (n = 2) were categorized as ST30, SCCmecIV, spa type 19, and PVL-positive, and as ST8, SCCmecIV, of a novel spa type 826, and PVL-negative. Eight different STs were detected among methicillin-susceptible S. aureus (MSSA) isolates, of which 55% were PVL-positive. One MSSA clone, which was categorized as ST121, spa type 273, and PVL-positive, caused fatal community-acquired pneumonia infections. The strains predominantly isolated in hospitals in Russia belonged to the multidrug-resistant Brazilian/Hungarian ST239 MRSA clone; however, this clone has new antibiotic susceptibilities. Additionally, the emergence of PVL-positive MSSA strains with enhanced virulence was observed, warranting continued surveillance.