No abstract

No abstract

Brucellosis is one of the leading causes of abortion in domestic animals that imposes costs on both economy and society. The disease is highly zoonotic and poses risk to animal handlers due to its zoonotic nature. It causes stillbirth, loss of kids and abortion in last term of pregnancy. Reproductive damage includes infertility in does and orchitis and epididymitis in breeding bucks, which result in high financial losses to farmers and the agriculture industry as a whole. It requires highly sensitive and specific assays to diagnose the disease at field level. In the current study, a visual loop-mediated isothermal amplification (LAMP) assay and the TaqMan® real-time PCR were developed with high sensitivity and specificity. For the TaqMan® probe, real-time PCR primers were developed using Omp31 gene as target and primers were designed using discontiguous conserved sequences of Omp31 gene. The Omp31 probes were designed by attaching 6-FAM reporter dye at the 5' end and BHQ-1 quencher at the 3' end. Published primers were used for visual LAMP assay targeting the Omp25 gene. Sensitivity of the standardized visual LAMP assay and TaqMan® real-time PCR assay was determined by serial dilution of positive Brucella melitensis DNA (10 to 10 ng) obtained from standard culture. The TaqMan® probe real-time assay can detect as low as 100 fg of B. melitensis DNA, whereas culture from vaginal swab washings has a limit of detection (LOD) of only 1 cfu/ml. Similarly, the visual LAMP assay can detect as low as 10 fg of B. melitensis DNA as compared to an LOD of 30 cfu/ml from culture of vaginal swab washings. Both assays were compared with serological tests (serum tube agglutination test (STAT) and indirect enzyme-linked immunosorbent assay (iELISA)) for diagnostic sensitivity and specificity. Diagnostic sensitivities and specificities for TaqMan® real-time PCR vs. LAMP assays were 98 and 100% vs. 100 and 97.8%, respectively. Results of visual LAMP assay indicated that LAMP is a fast, specific, sensitive, inexpensive and suitable method for diagnosis of B. melitensis infection under field conditions. On the other hand, Omp31 TaqMan® probe real-time assay can be used in conjunction with the other field-based diagnostic tests due to its high specificity.

The present study was conducted with the aim to record the pathological conditions in the female genital tract of goats. A total of 660 female genital tracts of goats were collected randomly from various slaughter houses as well as from CIRG post-mortem house regardless of breed and age (3 months to 8 years old) of the animals. Out of total 660 samples collected and examined, 154 (23.32%) revealed different types of pathological conditions. Highest occurrence of pathological conditions was observed in the uterus (53.24%), followed by those in the ovary (25.97%), cervix (11.03%), fallopian tubes (5.19%) and vagina (4.54%). Endometritis (37/154; 24.02%) was the most common pathological lesion followed by ovarian cyst (22/154; 14.28%). Other pathological conditions included congenital anomalies (3/154; 1.94%), hydrosalpinx (2/154; 1.29%), cervicitis (10/154; 6.4%), vaginitis (3/154; 1.94%) etc. Abnormalities like ovarian tumour, hydrometra, retention of placenta and uterine pigmentations were also recorded.In this study thelesions that could result into infertility or sterility of the goats occurred in 12.7% ofther eproductive systems examined.

In the present study, out of 1156 neonatal goat kids, 238 showing clinical diarrhea were used for detection of toxinotypes of Clostridium perfringens, enteropathogenic E. coli (ePeC), Group a rotavirus (GarV) and Bovine coronavirus (BCV). Isolation and toxinotyping of isolates were done by multiplex Polymerase chain reaction (PCr) using primers for cpa, cpb, cpb2, etx and iap genes. For EPEC, isolation and identification were done using bfpa gene and SYBr green based real time PCr (qPCr). GarV and BCV were detected, by onestep rt-PCr (osrt-PCr). the incidence of C. perfringens was 15.13% with 75% isolates toxinotype a, 25% type D and 61.11% of isolates carrying the β2-toxin gene. The incidence of EPEC was 68.07% based on qPCr, whereas 21.85% were positive for GarV and 15.97% for BCV by osrt-PCr. there was mixed infection of C. perfringens and ePeC in 11.76% and 3.78% for C. perfringens and GarV and 2.1% of C. perfringens and BCV. ePeC and GarV was 19.74% and ePeC plus BCV positivity was 11.34%. GarV and BCV was 5.88%, and 4.20% had mixed infection of ePeC, GarV and BCV. Of the total diarrheic kids sampled, 0.84% had mixed infection of C. perfringens, GARV, BCV and EPEC. On the basis of the above findings, it may be concluded that isolation, multiplex PCr and real time PCr facilitated the characterization of circulating C. perfringens toxinotypes and ePeC in goats reared under semi-arid conditions. the importance of enteritis caused by GarV and BCV and their role in mixed infection in goats requires extensive screening and pathogenicity studies to associate the symptoms with disease.