Molecular characterization of porcine reproductive and respiratory syndrome virus identified in 2021 from Nepal.

Porcine reproductive and respiratory syndrome (PRRS), an important viral disease of swine caused by PRRS virus (PRRSV) was first confirmed in Nepal in 2013. Since then, the virus has spread throughout the country and has now become endemic affecting the pig production nationally. However, molecular characterization of circulating strains has not been done in Nepal yet. In the present study, serum samples were collected from outbreak areas of different districts of Nepal and samples positive for PRRSV by ELISA were sent to Animal and Plant Health Agency (APHA), United Kingdom for sequence analysis. Out of 35 samples that were sent to APHA, only one sample was found positive by PCR and subjected to sequence analysis based on ORF5, ORF7 and Nsp2. The results from the phylogenetic analysis demonstrated that the PRRSV strain belongs to PRRSV-2 and lineage 8 strain. The sequences from the Nepalese PRRSV strain revealed a high degree of similarity with the strains isolated from India, China and Vietnam, with the closest genetic relatedness to the Indian isolates from 2020 and 2018. This is the first study on molecular characterization of PRRS virus circulating in Nepal. Further studies on strains circulating in Nepal are very essential to understand the virus diversity, its spread and evolution.

Farm characteristics and sero-prevalence of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in pigs of Nepal.

BACKGROUND: Porcine reproductive and respiratory syndrome is a highly infectious disease of swine caused by PRRS virus (PRRSV). OBJECTIVES: To evaluate the prevalence of PRRSV antibodies in the four districts of hilly and terai regions of Nepal. Toassess the farm characteristics through a questionnaire interview of farmersregarding management practices and PRRS. METHODS: A cross-sectional study was conducted from July 2020 to June 2021 to determine the sero-prevalence of PRRSV in pigs. A total of 180 porcine serum samples were collected from 23 pig farms and tested for PRRSV antibodies by ELISA. Alongside, farm characteristics were also assessed through questionnaire to determine the level of biosecurity measures in the farm, knowledge of the disease and possible control mechanisms. RESULTS: Out of 180 samples, 37 were tested positive resulting the overall sero-prevalence of 20.5%. There was significant association between different districts (p < 0.05) and PRRS prevalence. Prevalence of PRRSV antibody was found higher in Kaski district (10.5%) followed by Sunsari (8.8%) district. Based on age groups, highest prevalence was found in age groups of above 18 months (9.4%), followed by 13-18 months age groups (7.7%). Regarding the knowledge level of the disease, 43% of the farmers responded that they have heard about the disease. Biosecurity practices in the farm was found very poor where only 40% of the farms had disinfectant at the entrance of the farm and 25% pig farmers were found using separate boots while dealing with pigs. CONCLUSIONS: The findings of this study reveal the presence of PRRSV antibodies in pigs of Nepal. In addition poor biosecurity measures, management practices and poor knowledge level about the disease among farmers highly affect in the control and prevention of disease thereby affecting the pig production and productivity. Therefore, government should develop and implement effective control measures and biosecurity programs.

Lymphopenia Caused by Virus Infections and the Mechanisms Beyond.

Viral infections can give rise to a systemic decrease in the total number of lymphocytes in the blood, referred to as lymphopenia. Lymphopenia may affect the host adaptive immune responses and impact the clinical course of acute viral infections. Detailed knowledge on how viruses induce lymphopenia would provide valuable information into the pathogenesis of viral infections and potential therapeutic targeting. In this review, the current progress of viruses-induced lymphopenia is summarized and the potential mechanisms and factors involved are discussed.

Serological investigations of Peste des Petits Ruminants in cattle of Nepal.

Peste des Petits Ruminant (PPR) is an infectious viral disease of small ruminants caused by PPR virus. Although goat and sheep are the primary hosts of PPR, studies have continuously reported the prevalence of circulating antibodies in large ruminants, which could bring a potential challenge to effectively control and eradicate PPR. In Nepal, seroprevalence of PPRV antibodies in cattle have not been monitored yet. To address this, a total of 255 cattle sera were collected from Rupandehi, Banke, Bara and Chitwan districts of Nepal where outbreak of PPR in small ruminants was reported previously. The sera samples were tested by competitive ELISA and the result indicated the prevalence of 5.88% PPRV antibodies in cattle which indicates the exposure of cattle to PPR virus. To make the disease control program effective, intensive monitoring of both domestic and wild animals is very important.

Inhibition of eIF2α Phosphorylation by Peste des Petits Ruminant Virus Phosphoprotein Facilitates Viral Replication.

Peste des petits ruminant virus (PPRV) causes a highly contagious disease in small ruminants. The molecular mechanism of PPRV replication and its interactions with hosts are poorly studied. In other paramyxoviruses, the viral phosphoprotein (P) has been associated with multiple functions for key biological processes such as the regulation of transcription, translation, and the control of cell cycle. Phosphorylation of the α subunit of eukaryotic initiation factor 2 (eIF2α) is an important process for gene regulation in host cells under stress, including viral infection. In the present study, molecular mechanisms associated with PPRV replication and viral interaction with host cells were investigated. We describe the ability of PPRV to dephosphorylate eIF2α and the potential of PPRV P protein to induce the host cellular growth arrest DNA damage protein (GADD34), which is known to be associated with eIF2α dephosphorylation. Furthermore, we observed that PPRV P protein alone could block PERK/eIF2α phosphorylation. We speculate that PPRV exploits eIF2α dephosphorylation to facilitate viral replication and that PPRV P protein is involved in this molecular mechanism. This work provides new insights into further understanding PPRV pathobiology and its viral/host interactions.

Estimating Emission Load from Road Transportation within the Bhaktapur Municipality, Nepal.

Vehicular emissions have been playing a pivotal role in deteriorating air quality in many urban parts of Nepal causing adverse impacts upon the health of commuters and pedestrians attributed to severe respiratory diseases. Primary data such as the number of vehicles (N) were obtained using two-hour peak (8 am to 10 am) and two-hour nonpeak (1 pm to 3 pm) count, after which average annual vehicle kilometer (VKT) and fuel economy (F) required for emission load estimation were obtained from vehicle survey using the simple random sampling method, sampling size taken statistically under 5% margin of error. Secondary data in this study include emission factors and derived equations from a published article. The vehicular emission load of Bhaktapur Municipality were found to be 3,310 tons/year including CO2, CO, NOx, HC, and PM10 of which CO2 accounts for 94.36% of total emissions followed by CO (4.39%), HC (0.72%), NOx (0.35%), and PM10 (0.18%), respectively. Significant positive correlation was found (r = 0.92, p=0.002) between CO2 and PM10 (r = 0.87, p=0.009), between CO2 and NOx (r = 0.90, p=0.004), between CO and HC (r = 0.74, p=0.05), and between NOx and PM10, respectively. The scenario analysis shows that the introduction of electric vehicles at different rates within the municipality can reduce the emissions to a significant amount. Exponential growth in vehicular gaseous pollutants potent to jeopardize the environment and welfare can become inevitable in the future if clean energy technology is not promoted early.

Development of an Enzyme-Linked Immunosorbent Assay Based on CD150/SLAM for the Detection of Peste des Petits Ruminant Virus.

Peste des petits ruminant (PPR) is an economically important severe viral disease of small ruminants that affects primarily the respiratory and digestive tract. Specific detection of the PPR virus (PPRV) antigen plays an important role in the disease control and eradication program. In this study, an indirect enzyme-linked immunosorbent assay (ELISA) based on the recombinant goat signaling lymphocyte activation molecule (SLAM) as the capture ligand was successfully developed for the detection of the PPRV antigen (PPRV SLAM-iELISA). The assay was highly specific for PPRV with no cross-reactions among foot and mouth disease virus, Orf virus, sheep pox virus, and goat pox virus and had a sensitivity with a detection limit of 1.56 × 101 TCID50/reaction (50 µl). Assessment of 136 samples showed that the developed PPRV SLAM-iELISA was well correlated with real-time RT-qPCR assays and commercially available sandwich ELISA for detection of PPRV and showed relative sensitivity and specificity of 93.75 and 100.83%, respectively. These results suggest that the developed PPRV SLAM-iELISA is suitable for specific detection of the PPRV antigen. This study demonstrated for the first time that the goat SLAM, the cellular receptor for PPRV, can be used for the development of a diagnostic method for the detection of PPRV.

Expanding Diversity of Susceptible Hosts in Peste Des Petits Ruminants Virus Infection and Its Potential Mechanism Beyond.

Peste des petits ruminants (PPR) is a severe respiratory and digestive tract disease of domestic small ruminants caused by PPR virus (PPRV) of the genus Morbillivirus. Although the primary hosts of PPRV are goats and sheep, the host range of PPRV has been continuously expanding and reported to infect various animal hosts over the last decades, which could bring a potential challenge to effectively control and eradicate PPR globally. In this review, we focused on current knowledge about host expansion and interspecies infection of PPRV and discussed the potential mechanisms involved.

Host Cellular Receptors for the Peste des Petits Ruminant Virus.

Peste des Petits Ruminant (PPR) is an important transboundary, OIE-listed contagious viral disease of primarily sheep and goats caused by the PPR virus (PPRV), which belongs to the genus Morbillivirus of the family Paramyxoviridae. The mortality rate is 90-100%, and the morbidity rate may reach up to 100%. PPR is considered economically important as it decreases the production and productivity of livestock. In many endemic poor countries, it has remained an obstacle to the development of sustainable agriculture. Hence, proper control measures have become a necessity to prevent its rapid spread across the world. For this, detailed information on the pathogenesis of the virus and the virus host interaction through cellular receptors needs to be understood clearly. Presently, two cellular receptors; signaling lymphocyte activation molecule (SLAM) and Nectin-4 are known for PPRV. However, extensive information on virus interactions with these receptors and their impact on host immune response is still required. Hence, a thorough understanding of PPRV receptors and the mechanism involved in the induction of immunosuppression is crucial for controlling PPR. In this review, we discuss PPRV cellular receptors, viral host interaction with cellular receptors, and immunosuppression induced by the virus with reference to other Morbilliviruses.

Detection of antibodies against Bluetongue virus among domestic ruminants in the highlands of Nepal.

Bluetongue (BT) is one of the most economically important transboundary animal diseases. In recent years, it has been considered a disease related to climate change. A study was undertaken in 2013 in Nepal to measure the prevalence of Bluetongue virus (BTV) infection among domestic ruminants inhabiting the 3 agro-climatic zones with altitudes ranging from 150 to 2,400 metres above sea level. Twelve clusters representing the 3 altitudes were selected. The presence of antibodies against BTV was demonstrated in serum samples of sheep, goats, cattle, buffaloes, yaks/chauries, and chyangra goats (Himalayan goat) of Nepal. For this purpose, a total of 2,084 sera were collected from a population of 202 sheep, 739 goats, 590 cattle, 379 buffaloes, 105 yaks/chauries, and 69 chyangra goats between February 2013 and January 2014. The presence of antibodies against BTV was investigated using competitive enzyme-linked immunosorbent assay (c-ELISA). Of the 2,084 collected sera, 45.20% were positive for BTV antibodies. Species-wise prevalence was 17.82%, 47.50%, 53.05%, 58.05%, 7.62%, and 20.29% in sheep, goats, cattle, buffaloes, yak, and chyangra goats, respectively. Contrary to the general belief, maximum numbers of seropositive cases were recorded in buffaloes followed by cattle, goats, chyangra goats, sheep, and yak/chauries. The samples collected in the post-monsoon period (July-August is the monsoon period) show a seroprevalence higher than the pre-monsoon samples. This study shows the seroprevalence of BT in domestic ruminant population of Nepal at all altitudes. The highest prevalence has been reported in the plains of Terai followed by gradual decline in the mid-hills, and in the high mountains. Furthermore, detection of antibodies against BTV in both small and large ruminants (chyangra goats and yak/chauries) dwelling in high altitudes in the absence of BT vaccination is suggesting vector movement to the highlands as a consequence of warmer climate. These findings suggest that the climatic conditions, even at the higher elevation, are suitable for the survival of biting midges responsible for the transmission of BTV.