Nucleocapsid (N) protein gene of turkey coronavirus (TCoV) was expressed in a prokaryotic system and used to develop an enzyme-linked immunosorbent assay (ELISA) for detection of antibody to TCoV. Anti-TCoV hyperimmune turkey serum and normal turkey serum were used as positive or negative controls for optimization of the ELISA. Goat anti-turkey IgG (H+L) conjugated with horseradish peroxidase was used as detector antibody. Three hundred and twenty two turkey sera from the field were used to evaluate the performance of ELISA and determine the cut-off point of ELISA. The established ELISA was also examined with serum samples obtained from turkeys experimentally infected with TCoV. Those serum samples were collected at various time intervals from 1 to 63 days post-infection. The optimum conditions for differentiation between anti-TCoV hyperimmune serum and normal turkey serum were recombinant TCoV N protein concentration at 20 μg/ml, serum dilution at 1:800, and conjugate dilution at 1:10,000. Of the 322 sera from the field, 101 were positive for TCoV by immunofluorescent antibody assay (IFA). The sensitivity and specificity of the ELISA relative to IFA test were 86.0% and 96.8%, respectively, using the optimum cut-off point of 0.2 as determined by logistic regression method. Reactivity of anti-rotavirus, anti-reovirus, anti-adenovirus, or anti-enterovirus antibodies with the recombinant N protein coated on the ELISA plates was not detected. These results indicated that the established antibody-capture ELISA in conjunction with recombinant TCoV N protein as the coating protein can be utilized for detection of antibodies to TCoV in turkey flocks.
In this research Romanian Sheep pox virus was identified and confirmed by using PCR, Live attenuated Romanian Sheep pox vaccine (RSPV) was prepared and its titer on VERO cell was log 10 2.5 TCID50/dose. It was sterile, safe and potent. We used eight susceptible calves 6-8 months old, five calves vaccinated with 0.5 ml of prepared RSP vaccine intra dermally (I/D) in tail folds while three calves kept as control. Evaluation of both acquired humoral and cellular immunity by using lymphocyte proliferation assay, SNT, ELISA and Interferon Gamma Bioassay (IFN-ᵧ). The result showed that lymphocyte proliferation began to increase till reach to its peak (1.629) at 10 th day then decrease after that, While Interferon gamma (IFN-ᵧ) detected in 1 st day till 7 th day post vaccination then decreases after that. Results of SNT and ELISA revealed that Protective serum neutralizing antibody titer started at three weeks (1.5), (1.09) post vaccination then reach to its peak at 12 th weeks (2.5), (1.85) respectively and persisted till 28 weeks. From this study we found that live attenuated RSP vaccine a good immunogenic response where it was induced a high level of antibody titer with prolonged duration of immunity and higher lymphocyte and interferon gamma levels, So It considered good vaccine to control Lumpy Skin Disease (LSD).
Mastitis is a multifactorial disease and very difficult to control. It results from injury, chemical irritation and infection caused by different bacterial species. Mastitis remains one of the most common economic problems of dairy industry worldwide as it is the most expensive disease of dairy animals resulting in the reduction of milk production and quality. These expenses in terms of reduction of production, discarding milk, drug therapy, veterinarian charges, culling of incurable animals and extrause of labor. Analysis of bacteriological examination of milk samples was made to identify the main etiological agents involved in the disease. The organisms were identified on the basis of their cultural, staining characteristics, biochemical reactions .and molecular detection. Milk sample of 23 cows, which were positive for California mastitis test, cultured for microbiological examination in the study period. Two bacterial species were isolated, Staphylococcus aureus (Staph. aureus) and E. coli bacterial isolates. The predominant isolated bacteria were Staph. aureus with isolation rate of 37.77% however E. coli was isolated with isolation rate of 13.33%). Serum alkaline phosphatase (ALP) enzyme and calcium levels were highly significant decreased while C-Reactive protein (CRP) titre and phosphorus levels were highly significant increased. Lactate dehydrogenase enzyme(LDH), Aspartate aminotransferase enzyme(AST), Gamma-glutamyl transferase enzyme(GGT), Albumin, sodium, potassium and chloride levels were nonsignificantly changed in serum of mastitic cows compared to healthy ones. While LDH, ALP and phosphorus levels were highly significant increase in milk of mastitic cows compared to that of healthy ones. However, the calcium level was high significantly decreased in mastitic cows compared to healthy ones. Molecular detection of Staph. aureus and E. coli isolates by PCR. The expected sizes of PCR products of Staph. aureus was (984bp), while that for E. coli were (662bp).
Latex agglutination immunoassay is widely applied in diagnosis favored by advances in nanotechnology. This study was designed to investigate an inexpensive approach for producing highly specific protein antigen. Soluble Brucella proteins (SBPs 50) fractions were extracted from reference strain of Brucella abortus (strain 99) by differential precipitation with 50% ammonium sulfate. Analysis of the attained proteins by electrophoresis (SDS-PAGE) resulted in predominant 37.6 kDa protein, a medium-sized 30.6 kDa, and low molecular weight protein of 17.2 kDa when compared with low-molecular-mass marker ranged from (14 kDa to 92 kDa). To sustain as an advanced antigen applied in diagnosis of ruminant brucellosis, covalent coupling of obtained SBPs 50 to carboxylated polystyrene microsphere latex beads (0.81 ± 0.15 µm diameter) was used to assess the potential diagnostic utility of latex beads coated with the obtained SBPs. The developed assay (latex agglutination test, LAT) scored high diagnostic specificity (DSp) in all examined species; cattle, buffaloes, sheep, and goats (92.3%, 99.0%, 94.1%, and 95.9%) respectively. The diagnostic sensitivity (DSe) of LAT recorded (97.2%, 95.0%, 91.7%, and 85.6%) for cattle, buffaloes, sheep and goats correspondingly which were relatively lower than other counterpart conventional screening immunoassays; buffered antigens (BAPA and RBT). Kappa agreement values (ƙ) between latex agglutination test (LAT) and complement fixation test (CFT), were (0.90, 0.95, 0.83 and 0.76) for cattle, buffaloes, sheep and goats correspondingly indicating almost perfect correlation in examined species except in goats which showed substantial correlation. Besides DSe, DSp and ƙ agreement, other diagnostic indicators were assessed. From the results of this study, Latex agglutination test coated with SBPs 50 was proved to be an advisable method for diagnosis of ruminant brucellosis due to its high specificity, simplicity, fast, promptness, easy to interpret technique and low cost.
In this work five vaccination protocols were prepared and applied these protocols in sheep. Serum samples were collected from each group at time points 25, 50, 77, 89, and 98 days post vaccination. The results revealed that humoral antibodies were detected in groups 3, 4 and 5 by using Rose Bengal test (RBT), Buffer Acidified Plate Antigen Test (BAPAT), and Tube Agglutination Test at 98 days, 77 days, and 77 days respectively, while for ELISA test revealed that were positive at 50, 89, 98, 98, and 98 days post vaccination in group 1, 2,3,4 and 5 respectively. Cell mediated immunity was evaluated by Lymphocyte Blastogenesis Assay Test and Brucellin test (Delayed Type Hyper Sensitivity Test). The results indicated that there were no significant differences in between mean of different groups at P ≤ 0.05, So for Skin Delayed Hyper Sensitivity test, Group 1 and group 6 were negative while Group 2, 3, 4 and 5 were positive. Conclusion, animals in Groups 3, 4 and 5 had humoral immune response and can be protected from abortion in pregnant ewes and prevent infection. In this work, we evaluated to potential of three doses reduced Rev.1 mixed with E. coli flagellin which induced protection without need of adjuvant against I/P Brucella melitensis challenge. Also these data suggest that flagellin proteins might induce protective immune responses and these proteins will be a good candidate for subunit vaccine against ovine brucellosis in sheep.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.