Comparative and temporal transcriptome analysis of peste des petits ruminants virus infected goat peripheral blood mononuclear cells

Peste des petits ruminanats virus (PPRV), a morbillivirus causes an acute, highly contagious disease – peste des petits ruminants (PPR), affecting goats and sheep. Sungri/96 vaccine strain is widely used for mass vaccination programs in India against PPR and is considered the most potent vaccine providing long-term immunity. However, occurrence of outbreaks due to emerging PPR viruses may be a challenge. In this study, the temporal dynamics of immune response in goat peripheral blood mononuclear cells (PBMCs) infected with Sungri/96 vaccine virus was investigated by transcriptome analysis. Infected goat PBMCs at 48 h and 120 h post infection revealed 2540 and 2000 differentially expressed genes (DEGs), respectively, on comparison with respective controls. Comparison of the infected samples revealed 1416 DEGs to be altered across time points. Functional analysis of DEGs reflected enrichment of TLR signaling pathways, innate immune response, inflammatory response, positive regulation of signal transduction and cytokine production. The upregulation of innate immune genes during early phase (between 2-5 days) viz. interferon regulatory factors (IRFs), tripartite motifs (TRIM) and several interferon stimulated genes (ISGs) in infected PBMCs and interactome analysis indicated induction of broad-spectrum anti-viral state. Several Transcription factors – IRF3, FOXO3 and SP1 that govern immune regulatory pathways were identified to co-regulate the DEGs. The results from this study, highlighted the involvement of both innate and adaptive immune systems with the enrichment of complement cascade observed at 120 h p.i., suggestive of a link between innate and adaptive immune response. Based on the transcriptome analysis and qRT-PCR validation, an in vitro mechanism for the induction of ISGs by IRFs in an interferon independent manner to trigger a robust immune response was predicted in PPRV infection

Modulation of Host miRNAs Transcriptome in Lung and Spleen of Peste des Petits Ruminants Virus Infected Sheep and Goats

Peste des petits ruminants (PPR) is one of the highly contagious viral disease, characterized by fever, sore mouth, conjunctivitis, gastroenteritis, and pneumonia, primarily affecting sheep and goats. Reports suggested variable host response in goats and sheep and this host response vis-a-vis the expression of microRNAs (miRNAs) has not been investigated. Here, miRNAs were sequenced and proteomics data were generated to identify the role of differentially expressed miRNA (DEmiRNA) in PPR virus (PPRV) infected lung and spleen tissues of sheep and goats. In lungs, 67 and 37 DEmiRNAs have been identified in goats and sheep, respectively. Similarly, in spleen, 50 and 56 DEmiRNAs were identified in goats and sheep, respectively. A total of 20 and 11 miRNAs were found to be common differentially expressed in both the species in PPRV infected spleen and lung, respectively. Six DEmiRNAs—miR-21-3p, miR-1246, miR-27a-5p, miR-760-3p, miR-320a, and miR-363 were selected based on their role in viral infections, apoptosis, and fold change. The target prediction analysis of these six selected DEmiRNAs from the proteome data generated, revealed involvement of more number of genes in lung and spleen of goats than in sheep. On gene ontology analysis of host target genes these DEmiRNAs were found to regulate several immune response signaling pathways. It was observed that the pathways viz. T cell receptor signaling, Rap1 signaling, Toll-like receptor signaling, and B cell receptor signaling governed by DEmiRNAs were more perturbed in goats than in sheep. The data suggests that PPRV-induced miR-21-3p, miR-320a, and miR-363 might act cooperatively to enhance viral pathogenesis in the lung and spleen of sheep by downregulating several immune response genes. The study gives an important insight into the molecular pathogenesis of PPR by identifying that the PPRV—Izatnagar/94 isolate elicits a strong host response in goats than in sheep

Dysregulated miRNAome and Proteome of PPRV Infected Goat PBMCs Reveal a Coordinated Immune Response

In this study, the miRNAome and proteome of virulent Peste des petits ruminants virus (PPRV) infected goat peripheral blood mononuclear cells (PBMCs) were analyzed. The identified differentially expressed miRNAs (DEmiRNAs) were found to govern genes that modulate immune response based on the proteome data. The top 10 significantly enriched immune response processes were found to be governed by 98 genes. The top 10 DEmiRNAs governing these 98 genes were identified based on the number of genes governed by them. Out of these 10 DEmiRNAs, 7 were upregulated, and 3 were downregulated. These include miR-664, miR-2311, miR-2897, miR-484, miR-2440, miR-3533, miR-574, miR-210, miR-21-5p, and miR-30. miR-664 and miR-484 with proviral and antiviral activities, respectively, were upregulated in PPRV infected PBMCs. miR-210 that inhibits apoptosis was downregulated. miR-21-5p that decreases the sensitivity of cells to the antiviral activity of IFNs and miR-30b that inhibits antigen processing and presentation by primary macrophages were downregulated, indicative of a strong host response to PPRV infection. miR-21-5p was found to be inhibited on IPA upstream regulatory analysis of RNA-sequencing data. This miRNA that was also highly downregulated and was found to govern 16 immune response genes in the proteome data was selected for functional validation vis-a-vis TGFBR2 (TGF-beta receptor type-2). TGFBR2 that regulates cell differentiation and is involved in several immune response pathways was found to be governed by most of the identified immune modulating DEmiRNAs. The decreased luciferase activity in Dual Luciferase Reporter Assay indicated specific binding of miR-21-5p and miR-484 to their target thus establishing specific binding of the miRNAs to their targets.This is the first report on the miRNAome and proteome of virulent PPRV infected goat PBMCs

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Not AvailableMolecular identification of Peste-des-petits-ruminant virus from goats of assam state North-East India.Not Availabl

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Not Availablehe nucleotide sequences of the phosphoprotein (P) gene of peste des petits ruminants (PPRV) vaccine virus (PPRV Sungri/96) belongs to Asian lineage have been determined and the deduced amino acid sequences were compared with another vaccine strain PPRV/Nigeria75/1 and with those of the other morbilliviruses. The 1652 nucleotides of the P gene encode a phosphoprotein of 509 amino acid residues (from nucleotide numbers 60 to 1587), which is 91% identical to that of PPRV/Nigeria75/1. The C protein consists of 177 amino acid residues and is 91% identical with that of PPRV/Nigeria75/1. The conserved mRNA editing site (5′TTAAAAGGGCACAG) was present at positions 742–756 in the P gene, which is conserved in all other morbilliviruses. The CTT trinucleotide sequence is present at the N/P and P/M intergenic region, which is totally conserved in morbilliviruses. This will be the third sequence for the P gene of PPRV since that of the vaccine strain and a wild-type Turkish isolate has been published already.Not Availabl

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Not AvailableThis study describes the first confirmed report of contagious ecthyma in Black Bengal goats from Tripura state, a North-Eastern state of India situated at the Indo-Bangladesh border. Outbreaks were characterized by the high rates of morbidity (58-67%), low mortality (8-10%) and case fatality (11-15%). The etiology of the outbreaks was confirmed as orf virus (ORFV) by standard virological/serological and molecular techniques including sequence analysis of B2L, a major envelop protein gene of genus Parapoxvirus. Sequence and phylogenetic analysis based on B2L gene of ORFV isolates from Tripura revealed that they were closely related to each other and also to other Indian isolates, in particular to ORFV-Shahjahanpur 82/04 isolate from North India. They revealed several specific nucleotide/amino acid substitutions, namely G299A (G100D), G660A, C705T, C795T (N267D) and G872A (R291H) which may be of notable epidemiological significance. This report necessitates the systematic investigation of orf outbreaks in susceptible populations including wild species particularly at transboundary regions by use of rapid diagnostics to control the infection by deploying an effective vaccine/therapeutics and better managemental practices.Not Availabl

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Not AvailablePeste des petits ruminants (PPR) is a contagious, World Organization for Animal Health notifiable, economically important, transboundary morbilliviral disease of sheep and goats. Studying seroprevalence of PPR from different geographical areas under varying agro-climatic conditions may help in formulating effective and appropriate disease control strategies under the ongoing national PPR control program. The present cross-sectional study describes the prevalence of PPR virus antibodies in sheep and goats in the various epidemiological units in different states (Haryana, Himachal Pradesh [HP], Jammu and Kashmir [J&K], Punjab, Uttarakhand [UK], and Uttar Pradesh [UP]) of the northern region of India.Not Availabl

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Not AvailableThis cross-sectional serosurvey outlines the seroprevalence of peste des petits ruminants (PPR) in small ruminants in the hilly terrain north-eastern state of Sikkim in India. A total of 776 small ruminants’ (sheep [n = 99] and goats [n = 677]) serum samples were collected from 62 epidemiological units (epi-units) of the state using a two-stage sampling method from September 2017 to February 2018. The collected sera were screened for the small ruminant morbillivirus (SRMV) antibodies using a PPR-competitive enzyme-linked immunosorbent assay (cELISA). The results showed 1.16% (95% confidence interval [CI]: 0.61-2.19%) apparent prevalence with almost zero calculated true prevalence, which implies the insignificant prevalence of SRMV antibodies in the small ruminants population. Further, only a few samples from <5 % epi-units have shown positive for SRMV antibodies with an overall seropositive of <30% in the surveyed epi-units in Sikkim. This report is first of its kind on the seroprevalence of PPR in a unique niche of hilly terrain settings and it implies that SRMV has not yet become established in the small ruminants in Sikkim, despite the enzootic nature of the disease in the rest of India. This necessitates the imposing of strict quarantine measures and restrictions in the small ruminants trade along with the implementation of the intensive active surveillance programme to make a PPR free state.Not Availabl

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Not AvailableThe cross-sectional serosurvey for post-vaccination assessment of peste des petits ruminants (PPR) virus (PPRV) antibodies in sheep and goats was carried out in different states in the central and western regions of India after the implementation of vaccination under the PPR control programme. The serum samples (n = 4687) were collected from sheep (n = 1539) and goats (n = 3148) from August 2017 to March 2018 at various epidemiological units (n = 301) of the studied regions using a stratified random sampling method and PPR competitive ELISA kit was employed to detect PPRV antibodies. The results revealed 34, 21, 52, 74, 68, and 65% of prevalence of PPRV antibodies in small ruminants in Madhya Pradesh, Goa, Chhattisgarh, Maharashtra, Gujarat, and Rajasthan states, respectively, with a difference in seropositivity in sheep and goats across the states in sheep (p 50% prevalence of post vaccination PPRV antibodies across states due to variations in vaccination rates and patterns. The vaccination coverage and the reported outbreaks varied between the states in the studied regions. Due to continuous vaccination under the control program, the reported PPR outbreaks have progressively declined in most of the studied states, and the PPR risk areas are confined to a few districts and sporadically, outbreaks are reported indicating the effectiveness of vaccination. These findings provide valuable information on potential PPRV episystems, and will assist with activities regarding intensive surveillance, vaccination, biosecurity, and modification of policy decisions towards designing and implementing control and eradication measures. Further, the present situation necessitates continuous mass vaccination and active surveillance programs to make these regions free from PPR in consonance with the PPR Global Control and Eradication Strategy under the PPR Global Eradication Program.Not Availabl

Seroprevalence of peste des petits ruminants in sheep and goats in Eastern India

Not AvailableThe seroprevalence study of peste des petits ruminants (PPR) in small ruminants in Bihar and Odisha states in the Eastern region of India was carried out. A total of 1836 serum samples were collected from sheep (n = 648) and goats (n = 1188) from various epidemiological units (n = 112) in these states by a two-stage sampling plan during April 2017-March 2018. These samples were tested for the detection of virus antibodies by PPR competitive ELISA kit. The results revealed that the seroprevalence of PPR in sheep and goats in Bihar and Odisha states was 30.91% and 54.20%, respectively. Further, the chi-square analysis showed that the association exists between the presence of PPR virus antibodies in the goats (χ2 = 93.28, p < 0.01) and between the states (χ2 = 82.61, p < 0.01). This cross-sectional serosurvey also infers that the sheep and goats in most of the epi-units (n = 87) had < 70% of PPR virus antibodies prevalence. This warrants the intensive continuous mass vaccination program for a few more years to achieve the desired level of population immunity (epidemiological units protection level) and active surveillance to make these states free from PPR in the Eastern region of India.Not Availabl