Rhipicephalus appendiculatus serpin-3 (RAS-3), R. appendiculatus serpin-4 (RAS-4) and a 36kDa immuno-dominant protein of R. appendiculatus (RIM36) were reported as candidate antigens for the anti-tick vaccine to control ixodid ticks. In the present study, we generated recombinant proteins of RAS-3 (rRAS-3), RAS-4 (rRAS-4) and RIM36 (rRIM36), and assessed their potency as an anti-tick cocktail vaccine in cattle model. RT-PCR analysis showed that RAS-3, RAS-4 and RIM136 transcripts were detected in both adult male and female ticks during feeding. Immunization of cattle with the combination of rRAS-3, rRAS-4 and rRIM36 had raised antibodies against all recombinants and anti-sera had reacted with the molecules from the tick salivary gland extract. Tick infestation challenge demonstrated protective immunity against female ticks, resulting in mortality rates of 39.5 and 12.8 % for the vaccinated and control groups, respectively. Moreover, the mortality rate of Theileria parva-infected female ticks was 48.5 and 10.8 % in the vaccinated and control group, respectively. In order to evaluate the levels of pathogen transmission capacity by T. parva-infected ticks fed on immunized cattle, the occurrence of T. parva in the bovine parotid lymph node and peripheral blood was also determined and quantified by real-time PCR. Although the infection with T. parva could not be protected by the vaccine, the occurrence of pathogen in peripheral blood was delayed 1 to 2 days after the infestation challenge in vaccinated group. These results suggest that this cocktail vaccine plays a role in the prevention of tick infestation.

Puumala virus (PUUV), a causative agent of hemorrhagic fever with renal syndrome (HFRS), is prevalent in Europe and European Russia. No vaccine has been developed for PUUV-associated HFRS, primarily because of the low viral yield in cultured cells. A PUUV strain known as DTK/Ufa-97 was isolated in Russia and adapted for growth in Vero E6 cells maintained in serum-free medium. The DTK/Ufa-97 strain produced a higher viral titer in serum-free medium, suggesting that it may prove useful in the development of an HFRS vaccine. When PUUV-infected Vero E6 cells were grown in serum-free medium, the DTK/Ufa-97 strain yielded more copies of intracellular viral RNA and a higher viral titer in the culture fluid than did the Sotkamo strain. Phylogenetic analysis revealed that PUUVs can be classified into multiple lineages according to geographical origin, and that the DTK/Ufa-97 strain is a member of the Bashkiria-Saratov lineage. The deduced amino acid sequences of the small, medium, and large segments of the DTK/Ufa-97 strain were 99.2% to 100%, 99.3% to 99.8%, and 99.8% identical, respectively, to those of the Bashkirian PUUV strains and 96.9%, 92.6%, and 97.4% identical, respectively, to those of the Sotkamo strain, indicating that the PUUVs are genetically diverse. However, DTK/Ufa-97 and other strains of PUUV exhibited similar patterns of binding to a panel of monoclonal antibodies against Hantaan virus. In addition, diluted antisera (i.e., ranging from 1:160 to 1:640) specific to three strains of PUUV neutralized both homologous and heterologous viruses. These results suggest that the DTK/Ufa-97 strain is capable of extensive growth and is antigenically similar to genetically distant strains of PUUV.

An inactivated oil-based Newcastle disease vaccine was prepared using Mukteswar vaccine strain. The virus was propagated in 10-day old embryonating eggs and inactivated by 0.12% formalin for 48 hours at 37 degree C. The vaccine was formulated with 1 part antigen (aqueous phase) and 4 parts oil base. The oil base contained Tween-80 1%, Arlacel-A 10% and Mineral oil 89%. The stability of the vaccine was found satisfactory after 6 months and its viscosity and injectability was fairly ideal. The antigenicity of the vaccine was determined in 16 week-old pullets. The seromonitoring of the vaccinated and the control pullets was carried out for three months post- vaccination by Haemagglutination inhibition (HI) test. Blood samples were taken at fortnightly intervals. The Geometric mean HI titre of the vaccinated pullets on the day of vaccination 15, 30, 45, 60, 75, 90, 105 and 120 days post- vaccination was 18.4, 4.9, 87.5, 192.3, 257.6, 111.4, 91.7, 63.9 and 30.0. However, in non-vaccinated control pullets it was found to be 18.4, 3.7, 3.7, 4.3, 3.8, 4.0, 3.5, 2.8 and 2.3 respectively. The inactivated oil-based vaccine induced a marked antibody response which continued upto three months.

The level of antibody by ELISA,immunsosuppressive effect baes on the response of birds to Newcastle disease vaccination and damage to bursa of Fabricius by IBD vaccination were studied. The efficacy of six different IBD vaccination schedules were studied using intermediate and intermediate plus strains of vaccines either alone or in combination. In vaccinated groups, the sero-conversion of the vaccine virus was noticed during fourth week, reaching to the peak between eight to twelve weeks of age in different groups. Afterwards, there was a gradual decrease in the titres, by the end of 20th week (maximum period tested). There was no significant difference in the titres of different treatment groups. However, all the groups showed titres above protective level during the entire period of study. There was significant difference in bursa body weight (B-BW) ratios of vaccinated groups in comparison with control group. Histopathological studies of bursal sections revealed depletion of lymphoid follicles, presence of cystic spaces, edema and hemorrhages. The birds vaccinated with hot strain of IBD vaccine showed metaplastic changes, presence of foam cells with pronounced interfollicular fibrosis. The bursal scores were maximum in the groups vaccinated with hot strains of IBD vaccines.

This study was conducted to determine the optimum storage temperature for Rinderpest vaccine prepared on vero cells to know the shelf life of the vaccine. The vials were randomly selected from one batch of the vaccine, titrated and stored at minus 20 degree centigrade (Freezer), 4 degree centigrade (refrigerator) and room temperature. The titre was found to be 105.1 per ml. The vials stored at minus 20 degree centigrade & 4 degree centigrade were subjected to titration after an interval of six months for 3 and 2 years respectively. The vials stored at room temperature were tested after 4 weeks, 8 weeks and 12 weeks. Titration results indicated that the titre of the vaccine vials stored at room temperature decreased by 100.9 101.2 and 101.6 after storage time of 4 weeks, 8 weeks and 12 weeks respectively. The vials stored at 4 degree centigrade maintained their titre for a period of six months but after that the loss in titre was 100.4, 101.0, and 102.4 after storage time of one, one and half and two 2 years respectively. The vaccine vials stored at minus 20 degree centigrade maintained their original titres (initial titre of the vaccine) even after the storage for three years. It is concluded that vero cell adapted Rinderpest virus vaccine can be stored at 4 degree centigrade for a period of six months, however, at 20 degree centigrade it can be stored for three years without any adverse effect on titre.