Stability and integrity of self-assembled bovine parvovirus virus­like particles (BPV­VLPs) of VP2 and combination of VP1VP2 assisted by baculovirus-insect cell expression: a potential logistical platform for vaccine deployment.

BACKGROUND: Bovine parvovirus (BPV) is an autonomous DNA virus with a smaller molecular size and subtle differences in its structural proteins, unlike other animal parvoviruses. More importantly, this virus has the potential to produce visible to silent economic catastrophes in the livestock business, despite receiving very little attention. Parvoviral virus-like particles (VLPs) as vaccines and as logistical platforms for vaccine deployment are well studied. However, no single experimental report on the role of VP1 in the assembly and stability of BPV-VLPs is available. Furthermore, the self-assembly, integrity and stability of the VLPs of recombinant BPV VP2 in comparison to VP1 VP2 Cap proteins using any expression method has not been studied previously. In this study, we experimentally evaluated the self-assembling ability with which BPV virus-like particles (VLPs) could be synthesized from a single structural protein (VP2) and by integrating both VP2 and VP1 amino acid sequences. METHODS: In silico and experimental cloning methods were carried out. His-tagged and without-His-tag VP2 and V1VP2-encoding amino acid sequences were cloned and inserted into pFastbacdual, and insect cell-generated recombinant protein was evaluated by SDS‒PAGE and western blot. Period of infectivity and expression level were determined by IFA. The integrity and stability of the BPV VLPs were evaluated by transmission electron microscopy. The secondary structure of the BPV VLPs from both VP2 and V1VP2 was analyzed by circular dichroism. RESULTS: Our findings show that VP2 alone was equally expressed and purified into detectable proteins, and the stability at different temperatures and pH values was not appreciably different between the two kinds of VLPs. Furthermore, BPV-VP2 VLPs were praised for their greater purity and integrity than BPV-VP1VP2 VLPs, as indicated by SDS‒PAGE. Therefore, our research demonstrates that the function of VP1 has no bearing on the stability or integrity of BPV-VLPs. CONCLUSIONS: In summary, incredible physiochemically stable BPV VP2-derived VLPs have been found to be promising candidates for the development of multivalent vaccines and immunodiagnostic kits against enteric viruses and to carry heterogeneous epitopes for various economically important livestock diseases.

Prioritization of neglected tropical zoonotic diseases: A one health perspective from Tigray region, Northern Ethiopia.

Neglected tropical zoonotic diseases (NTZDs) continue to have a major effect on the health of humans and animals. In this study, a one health approach was used to prioritize and rank neglected tropical zoonotic diseases at the regional and zonal levels in Tigray National Regional State, Ethiopia. For prioritization of NTZDs a cross-sectional study through a structured questionnaire was administered to 313 health experts from human and animal health sectors. In addition, focus group discussions (FGD) were held with purposively selected key informants. Descriptive, and Multivariable analysis was applied to report the results and a ranked list of diseases was developed at the zonal and regional level. In the region, 8 of the 12 World Health Organization listed NTZDs were considered major diseases including anthrax, brucellosis, bovine tuberculosis, taeniasis, leishmaniasis, rabies, schistosomiasis, and soil-transmitted helminths. Considering the zoonotic and socioeconomic importance of the diseases at the regional level, rabies ranked 1stwhereas anthrax, bovine tuberculosis, leishmaniasis, and brucellosis were ranked from 2nd to 5th, respectively. The FGD result also supported the prioritization result. The Multivariable analysis showed a statistically significant difference in the zonal distribution of anthrax (р = 0.009, OR = 1.16), taeniasis (p<0.001, OR = 0.82), leishmaniasis (p<0.001, OR = 1.91), rabies (p = 0.020, OR = 0.79) and soil-transmitted helminths (p = 0.007, OR = 0.87) but not for brucellosis (p = 0.585), bovine tuberculosis (p = 0.505), and schistosomiasis (p = 0.421). Anthrax (p<0.001, OR = 26.68), brucellosis (p<0.001, OR = 13.18), and taeniasis (p<0.001, OR = 6.17) were considered as the major zoonotic diseases by veterinary practitioners than human health practitioners whereas, leishmaniasis was recognized as a major health challenge by human health professionals. Understanding the priority diseases in the region is supportive for informed decision-making and prioritizes the limited resources to use. Furthermore, strengthening the collaboration between human and animal health professions is important to control the diseases.