Molecular characterization of pathogenic Escherichia coli isolated from diarrheic and in-contact cattle and buffalo calves.

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 (stx1 and stx2), 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. coli (EAEC) with a 30.7, 2.7, 12.7, and 7.3% distribution, respectively. Meanwhile, the detection rates of f5, stx1, stx2, st, lt, eae, hylA, and astA genes were 17.3, 27.3, 6.7, 10, 37.3, 17.7, 9.3, and 20.7%, respectively. These virulence genes were found either single or in different combinations, such as stx/eae, stx/st/f5, eae/st/f5, or st/lt/f5. Four attaching-effacing shigatoxigenic E. coli isolates (AE-STEC) harboring stx/eae were retrieved from diarrheic calves. Although none of the stx-or eae-positive isolates was verified as O157:H7, STEC isolates detected in apparently healthy calves have potential pathogenicity to humans highlighting their zoonotic importance as reservoirs. Atypical combinations of ETEC/STEC and ETEC/EPEC were also detected in percentages of 14.7 and 2.7%, respectively. Most of these atypical combinations were found more in buffalo calves than in cattle calves. While STEC and EPEC isolates were detected more in cattle calves than in buffalo calves, ETEC isolates were the same in the two species. The pathogenic E. coli infection in calves was recorded to be higher in the first weeks of life with the largest numbers of virulence factor-positive isolates detected at the age of 4 weeks. Histopathological examination of five intestinal samples collected from four dead buffalo calves revealed typical attaching and effacing (AE) lesion which was correlated with the presence of intimin encoding virulence gene (eae). Other lesions characterized by hemorrhagic enteritis, shortening and fusion of intestinal villi and desquamation of the lining epithelium of intestinal mucosa had also been detected.

Serological evaluation for the current epidemic situation of foot and mouth disease among cattle and buffaloes in Egypt.

AIM: The present study was aimed to investigate the epidemic situation of foot-and-mouth disease (FMD) in Egypt from 2016 to 2018 based on the detection of FMD virus (FMDV) in carrier or previously infected animals, by determination of antibodies against non-structural protein (NSP), implementation a pilot study on circulating FMDV serotypes and assure the efficacy of locally produced inactivated trivalent vaccine. MATERIALS AND METHODS: A total of 1500 sera were collected from apparent healthy vaccinated cattle and buffaloes from three Egyptian geographical sectors, representing ten governorates. Determination of FMD antibodies against NSP was carried out using 3ABC enzyme-linked immunosorbent assay (ELISA) test. Serotyping of the circulating FMDV and assure the vaccine efficacy was performed using solid-phase competitive ELISA. RESULTS: The 3ABC ELISA test revealed 26.4% and 23.7% positive for FMDV-NSP antibodies in cattle and buffalo sera, respectively. The highest positivity was in Delta Sector among both cattle 42.3% and buffaloes 28.8%. Serotyping of FMDV-positive NSP sera in El-Qalyubia Governorate for the circulating FMDV serotypes O, A, and Southern African Territories (SAT) 2 was 52.2%, 17.4%, and 30.4% in cattle and 31.8%, 27.3%, and 40.9% in buffaloes, respectively. The overall protection level due to the vaccination program was 62.1 and 60.9% in cattle and buffaloes, respectively, while the protective level of the FMDV serotypes O, A, and SAT2 included in the inactivated trivalent vaccine was 73.9, 84.6, and 63.8% in cattle and 72.3, 82.3, and 63.5% in buffaloes, respectively. CONCLUSION: The present study recommended full determination for the immunogenic relationship between the vaccine strains and the field strains to attain maximum protection against the circulating viruses.

Evaluation of different diagnostic methods for diagnosis of Lumpy skin disease in cows.

Viral isolation, polymerase chain reaction (PCR), dot blot hybridization (DBH), and indirect enzyme-linked immunosorbent assay (iELISA) were used for the diagnosis of lumpy skin disease in clinically infected, fevered, and apparently normal dairy cows. Lumpy skin disease virus (LSDV) was isolated from skin biopsies and blood samples collected from clinically infected cows in percentages of 72% and 20%, respectively. The virus recovered from blood samples collected from fevered cows in percentage of 33.3%. Both PCR and DBH detected viral DNA in 100% of skin biopsies collected from clinically infected cows whereas the detection rates in blood samples collected from clinically infected animals were 100% and 84% using PCR and DBH, respectively. Viral DNA was detected in blood samples collected from fevered cows using PCR and DBH in percentages of 77.8% and 66.6%, respectively. Only 19.1% of blood samples collected from in-contact cows was positive for both of PCR and DBH. Detection rates of antibodies against LSDV using iELISA in serum samples collected from clinically infected and fevered cows were 56% and 11.1%, respectively, whereas all in-contact cows had no antibodies against the virus.