Ruminant Morbillivirus Research Articles

Development of a SYBR Green-based RT-qPCR assay for the rapid detection of peste-des-petits-ruminants virus.

Peste-des-petits-ruminants (PPR) is primarily a disease of small ruminants caused by peste-des-petits-ruminants virus (PPRV; Paramyxoviridae, Morbillivirus caprinae), formerly the small ruminant morbillivirus. PPRV can cause significant morbidity and mortality in small ruminants and a significant economic impact. Conventional reverse-transcription PCR (RT-PCR), and probe-based and SYBR Green-based RT quantitative real-time PCR (RT-qPCR), are employed for the molecular detection of PPRV. Here we describe a SYBR Green-based RT-qPCR for rapid and sensitive detection of PPRV. We designed the specific primers from the conserved region of the fusion gene (F) of PPRV. The standard curve of the established RT-qPCR assay had a good linear relationship. The developed assay was also 3 log units more sensitive than the conventional RT-PCR, with a detection limit of 13.6 copies and an efficiency of 98.2%. There was no cross-reactivity with other caprine respiratory viruses, namely bluetongue virus, goatpox virus, and orf virus. The positive detection rate of clinical samples was 11 of 64 (17.2%) versus 10 of 64 (15.6%) by conventional RT-PCR. We confirmed our results by sequencing the full F and N genes of the isolates. Our SYBR Green RT-qPCR can be used as a fast, economical, and sensitive alternative to RT-PCR for the detection of PPRV.

Molecular epidemiology of small ruminant morbillivirus (SRMV) isolates from field outbreaks in Kerala, India based on fusion (F) and nucleoprotein (N) gene

Molecular epidemiology of small ruminant morbillivirus (SRMV) isolates from field outbreaks in Kerala, India based on fusion (F) and nucleoprotein (N) gene

Development and characterization of a novel nanobody with SRMV neutralizing activity

Peste des petits ruminants (PPR) is an acute, contact infectious disease caused by the small ruminant morbillivirus (SRMV), and its morbidity in goats and sheep can be up to 100% with significant mortality. Nanobody generated from camelid animals such as alpaca has attracted wide attention because of its unique advantages compared with conventional antibodies. The main objective of this study was to produce specific nanobodies against SRMV and identify its characteristics. To obtain the coding gene of SRMV-specific nanobodies, we first constructed an immune phage-displayed library from the VHH repertoire of alpaca that was immunized with SRMV-F and -H proteins. By using phage display technology, the target antigen-specific VHHs can be obtained after four consecutive rounds of biopanning. Results showed that the size of this VHH library was 2.26 × 1010 CFU/mL and the SRMV-F and -H specific phage particles were greatly enriched after four rounds of biopanning. The positive phage clones were selected and sequenced, and total of five independent different sequences of SRMV-specific nanobodies were identified. Subsequently, the DNA fragments of the five nanobodies were cloned into E. coli BL21(DE3), respectively, and three of them were successfully expressed and purified. Specificity and affinity towards inactivated SRMV of these purified nanobodies were then evaluated using the ELISA method. Results demonstrated that NbSRMV-1-1, NbSRMV-2-10, and NbSRMV-1-21 showed no cross-reactivity with other antigens, such as inactivated BTV, inactivated FMDV, His-tag labeled protein, and BSA. The ELISA titer of these three nanobodies against inactivated SRMV was up to 1:1000. However, only NbSRMV-1-21 displayed SRMV neutralizing activity at a maximum dilution of 1:4. The results indicate that the nanobodies against SRMV generated in this study could be useful in future applications. This study provided a novel antibody tool and laid a foundation for the treatment and detection of SRMV.

Determining the decreasing trend of maternal immunity against small ruminant morbillivirus in goat kids.

Small ruminant morbillivirus (SRMV) is the etiological agent of Peste des petits ruminants (PPR) disease. PPR is one of the most important viral diseases of small ruminant husbandry. In the endemic countries, vaccination is the main way to control this disease. Administering the first PPR vaccine in goat kids requires decreased maternal immunity. The aim of this study was to determine the decreasing trend of maternal immunity against SRMV in goat kids born from vaccinated goats. Twenty Saanen goat kids were studied in two groups including control (n = 5, receiving colostrum from unvaccinated goats) and treatment (n = 15, receiving colostrum from vaccinated goats). Virus neutralisation (VN) test was used to evaluate serum specific antibodies against SRMV in goat kids from birth to 100 days of age. The first goat kid (n = 1) in the treatment group was seronegative at the age of 28 days. All the goat kids were seronegative at the age of 100 days. The average serum titre of the goat kids at the age of 70-100 days became negative. Some goat kids became seronegative before reaching the age of receiving the first PPR vaccine. The age of 70-100 days could be a good age range to give the first dose of PPR vaccine to the goat kids, but more studies were needed on the effectiveness of this vaccine at this age range.

Serological Detection of SRMV, Bovine Viral Diarrhea Virus, BVDV and BEFV in Camels (Camelus dromedarius) in Southwest Iran

Background: Small ruminant morbillivirus (SRMV), bovine viral diarrhea virus (BVDV), bovine herpes virus-1 (BHV-1), and bovine ephemeral fever virus (BEFV) are among the most important viruses of farm animals. Objectives: This study aimed at the serological detection of SRMV, BVDV, BHV-1, and BEFV in the camel population of Khuzestan Province, located in southwest Iran. Methods: A total of 150 camel blood samples were randomly collected from free-ranging, seemingly healthy camels of both sexes and various ages in 8 different regions of Khuzestan Province, Iran. The sera were tested for SRMV, BVDV, BHV-1, and BEFV with serum neutralization test. Results: The seropositive samples for SRMV, BVDV, and BHV-1 were 1(0.67%), 7(4.67%), and 11(7.33%), respectively. There was no seroconversion against BEFV in the serum samples. Conclusion: The camel population of Khuzestan Province is subclinically infected with SRMV, BVDV, and BHV-1 and could play a significant role in the epizootiology of these viral diseases in this region, which is very important for the control and eradication programs.

Immunoinformatics Approach for Epitope Mapping of Immunogenic Regions (N, F and H Gene) of Small Ruminant Morbillivirus and Its Comparative Analysis with Standard Vaccinal Strains for Effective Vaccine Development

Outbreaks of small ruminant morbillivirus (SRMV) are regularly occurring in Pakistan despite vaccine availability. This study was designed to identify substitutions within the immunogenic structural and functional regions of the nucleocapsid, fusion, and hemagglutinin genes of SRMV and their comparison with vaccinal strains of Nigerian and Indian origin. Swabs and tissue samples were collected from diseased animals. RT-PCR was used to characterize selected genes encoded by viral RNA. The study's N, F, and H protein sequences and vaccinal strains were analyzed for B and T cell epitope prediction using ABCpred, Bipred, and IEDB, respectively. Significant substitutions were found on the C terminus of the nucleocapsid, within the fusion motif region of the fusion gene and in the immunoreactive region of the hemagglutinin gene. Our results emphasize the need for the development of effective vaccines that match the existing variants of SRMV strains circulating in Pakistan.

Comparison of Biosafety and Diagnostic Utility of Biosample Collection Cards.

Six different biosample collection cards, often collectively referred to as FTA (Flinders Technology Associates) cards, were compared for their ability to inactivate viruses and stabilize viral nucleic acid for molecular testing. The cards were tested with bluetongue virus, foot-and-mouth disease virus (FMDV), small ruminant morbillivirus (peste des petits ruminants virus), and lumpy skin disease virus (LSDV), encompassing non-enveloped and enveloped representatives of viruses with double-stranded and single-stranded RNA genomes, as well as an enveloped DNA virus. The cards were loaded with virus-containing cell culture supernatant and tested after one day, one week, and one month. The inactivation of the RNA viruses was successful for the majority of the cards and filters. Most of them completely inactivated the viruses within one day or one week at the latest, but the inactivation of LSDV presented a greater challenge. Three of the six cards inactivated LSDV within one day, but the others did not achieve this even after an incubation period of 30 days. Differences between the cards were also evident in the stabilization of nucleic acid. The amount of detectable viral genome on the cards remained approximately constant for all viruses and cards over an incubation period of one month. With some cards, however, a bigger loss of detectable nucleic acid compared with a directly extracted sample was observed. Using FMDV, it was confirmed that the material applied to the cards was sufficiently conserved to allow detailed molecular characterization by sequencing. Furthermore, it was possible to successfully recover infectious FMDV by chemical transfection from some cards, confirming the preservation of full-length RNAs.

Novel and classical morbilliviruses: Current knowledge of three divergent morbillivirus groups.

Currently, seven species of morbillivirus have been classified. Six of these species (Measles morbillivirus, Rinderpest morbillivirus, Small ruminant morbillivirus, Canine morbillivirus, Phocine morbillivirus, and Cetacean morbillivirus) are highly infectious and cause serious systemic diseases in humans, livestock, domestic dogs, and wild animals. These species commonly use the host proteins signaling lymphocytic activation molecule (SLAM) and nectin-4 as receptors, and this usage contributes to their virulence. The seventh species (Feline morbillivirus: FeMV) is phylogenetically divergent from the six SLAM-using species. FeMV differs from the SLAM-using morbillivirus group in pathogenicity and infectivity, and is speculated to use non-SLAM receptors. Recently, novel species of morbilliviruses have been discovered in bats, rodents, and domestic pigs. Because the ability to use SLAM and nectin-4 is closely related to the infectivity and pathogenicity of morbilliviruses, investigation of the potential usage of these receptors is useful for estimating infectivity and pathogenicity. The SLAM-binding sites in the receptor-binding protein show high similarity among the SLAM-using morbilliviruses. This feature may help to estimate whether novel morbillivirus species can use SLAM as a receptor. A novel morbillivirus species isolated from wild mice diverged from the classified morbilliviruses in the phylogenetic tree, forming a third group separate from the SLAM-using morbillivirus group and FeMV. This suggests that the novel rodent morbillivirus may exhibit a different risk from the SLAM-using morbillivirus group, and analyses of its viral pathogenicity and infectivity toward humans are warranted.

Integrated analysis of long-noncoding RNA and circular RNA expression in Peste-des-Petits-Ruminants Virus (PPRV) infected marmoset B lymphocyte (B95a) cells.

Peste-des-Petits-Ruminants (PPR) or goat plague is an important viral disease of sheep and goats caused by the small ruminant morbilli virus or PPR virus (PPRV). Long non coding RNAs (lncRNA) and circular RNAs (circRNA) play a pivotal role in several biological processes including regulation of virus-host interactions. The present study explored the expression of lncRNA, circRNA and their functions in PPRV infected B-lymphocyte (B95a) cells. The results revealed a total of 4531 lncRNA and 2348 circRNA expression in both mock and PPRV infected samples. Analysis of differentially expressed (DE) RNA identified 123 DE-lncRNA and 39 DE-circRNA as significantly dysregulated. Functional analysis of cis-target genes of DE-lncRNA indicated activation of TCF dependent WNT signaling and PKN1 stimulated transcription process. Interactions (sponging) of microRNA (miRNA) revealed 344 DE-lncRNA-miRNA and 93 DE-circRNA-miRNA pairs. The competing endogenous RNA (ceRNA) network of lncRNA/circRNA-miRNA-mRNA in PPRV infected B95a cells was represented by 69 ceRNA pairs. We validated the DE-circRNA by targeted amplification and sequencing of back spliced junctions (BSJs). The present study revealed a profile of lncRNA, circRNA and their potential ceRNA network in PPRV infection. The results provide insight for better understanding of PPRV-host interactions.

The extraction of Peste Des Petits Ruminants Virus RNA from paraffin-embedded tissues using a modified extraction method

Peste des petits ruminants (PPR) which is caused by small ruminant morbillivirus (PPRV) has an important economic impact on small ruminant farming due to high mortality rates, weight loss and restrictions on the export of small ruminants products. Molecular assays are commonly used in the diagnosis of the disease. Extraction of RNA from formalin-fixed paraffin-embedded (FFPE) tissues is challenging because of the RNA is often degraded by formalin fixation process. Although commercial kits have been developed for extraction of nucleic acids from FFPE tissues, they are expensive than other extraction kits. In this study, a modified extraction method was evaluated for detection of PPRV from FFPE tissues. A total of 20 FFPE tissue samples including 15 PPRV positive and 5 PPRV negative FFPE tissue samples were used. Two years ago, these selected FFPE tissue samples were analysed by nucleoprotein gene based one step real time RT-PCR method before they were fixed with formalin and embedded in paraffin. FFPE tissue samples were extracted using modified extraction method and were tested by fusion (F) gene based one step RT-PCR. PPRV specific RNA was detected in 12 FFPE tissue samples whereas 3 positive samples were found negative by one-step RT-PCR. Furthermore, 5 negative FFPE tissue samples were also found negative. Three false negative results were from samples with high real-time RT-PCR cycle threshold. Therefore, false negative results could be related with lower viral loads which might be lower than detection limit of the one-step RT-PCR. The results of the study show that modified extraction method could be used for RNA extraction from FFPE tissues which had been stored for 2 years.

The Experimental Infection of Goats with Small Ruminant Morbillivirus Originated from Barbary Sheep.

Peste des Petits Ruminants (PPR) is a transboundary contagious disease in domestic small ruminants. Infections with the small ruminant morbillivirus (SRMV) were regularly found in wildlife, with unknown roles in PPR epidemiology. In order to access infection dynamics and virulence, we infected German Edelziege goats intranasally with a SRMV isolate that originated from Barbary sheep from an outbreak in the United Arab Emirates. Six goats were infected with cell culture-isolated SRMV, and two goats were kept in contact. Goats were daily monitored, and clinical score was recorded. EDTA blood, nasal, conjunctival and rectal swab samples were collected for the detection of SRMV genome load and serum for serological analysis. Short incubation period in infected (4 to 5 dpi) as well as in contact goats (9 dpi) was followed by typical clinical signs related to PPR. The highest viral load was detectable in conjunctival and nasal swab samples with RT-qPCR and rapid pen-side test. Specific antibodies were detected at 7 dpi in infected and 14 dpi in contact goats. In general, high virulence and easy transmission of the virus originated from wildlife in domestic goats was observed. The virus isolate belongs to Asian lineage IV, genetically related to Chinese and Mongolian strains.

Epidemiology, risk factors and molecular characterization of small ruminant morbillivirus in Haryana, India

Peste des petitis ruminants is an economically important transboundary and notifiable viral disease of sheep and goats. In this study, 14 PPR suspected outbreaks among sheep and goats were investigated in four districts of Haryana, India, during July 2020 to October, 2021. The causative agent of the disease; small ruminant morbillivirus was detected by Reverse Transcription Polymerase Chain Reaction targeting full gene sequences of fusion protein gene and confirmed by sequencing. The overall morbidity and cumulative mortality in these outbreaks were 37.56% and 12.09%, respectively. Risk factor analysis identified significant difference in mortality based on age with higher mortality in young ones; 21% as compared to adults; 7.55%. Analysis of the vaccination status revealed significant difference in morbidity and mortality with higher morbidity and mortality in un-vaccinated animals as compared to vaccinated ones. The sequencing and phylogenetic analysis of representative samples revealed that the strains of the present study fall in lineage IV (96.6–99.1%) along with other Indian isolates but made a separate cluster (sub-lineage). The comparison of deduced amino acid (aa) sequence analysis of fusion protein of circulating field strains with reference vaccine strain and other lineage IV strains revealed four N-linked glycosylation sites instead of three. The findings of the present study revealed changes in fusion protein of some of the circulating field strains of SRMV in Haryana, India. Further detailed studies are warranted to delineate the molecular details of these circulating field strains and to evaluate the effectiveness of currently used vaccine against these mutated strains.

Prevalence, Risk Factors for Exposure, and Socio-Economic Impact of Peste Des Petits Ruminants in Karenga District, Karamoja Region, Uganda.

Peste des petits ruminants (PPR), a disease caused by small ruminant morbillivirus (SRM), is highly contagious with high morbidity and mortality. Controlling PPR requires a proper understanding of the epidemiological dynamics and impact of the disease in a range of geographical areas and management systems. Karenga district, located in the pastoral region of Karamoja in northeastern Uganda, and in the vicinity of Kidepo Valley National Park, is characterised by free cross-border (South Sudan and Kenya) livestock trade, communal grazing, and transhumance. This study was conducted from November through December 2020 to determine the seroprevalence of anti-SRM antibodies, the risk factors associated with the occurrence, and the socio-economic impact of PPR in Karenga. A total of 22 kraals were randomly selected from all administrative units, and 684 small ruminants (sheep = 115, goats = 569) were selected for serum collection using systematic random sampling. Exposure to SRM was determined using a competitive enzyme-linked immunosorbent assay. The overall true seroprevalence of SRM antibodies was high, 51.4 (95% confidence interval [CI] 45–52.6). Multivariate logistic regression for risk factors showed that seroprevalence varied significantly by location (26.8% to 87.8%, odds ratio (OR) ≤ 14.5). The odds of exposure to SRM were higher in sheep (73.9%) than in goats (43.8%) (OR = 1.7, p = 0.08), and seropositivity was higher in animals greater than two years old (65.5%; OR = 11.1, p < 0.001), or those one to two years old (24.7%; OR = 1.6, p = 0.2), compared to small ruminants less than one year old (16.1%). Using participatory epidemiology approaches (semi-structured interviews, clinical examinations, pairwise ranking, proportional piling, impact matrix scoring) with 15 key informants and 22 focus groups of pastoralists, PPR was the second most important small ruminant disease: relative morbidity 14%, relative mortality 9%, and case fatality rate 78%, and impacted productivity mainly in terms of treatment costs, mortality, marketability, and conflicts. These findings provide evidence to support the implementation of disease surveillance and control strategies to mitigate the impact of PPR in Karamoja and other pastoral areas in eastern Africa.

Mutation Profiles of eGFP-Tagged Small Ruminant Morbillivirus During 45 Serial Passages in Ribavirin-Treated Cells.

Small ruminant morbillivirus (SRMV), formerly known as peste-des-petits-ruminants virus, classified into the genus Morbillivirus in the family Paramyxoviridae. Its L protein functions as the RNA-dependent RNA polymerases (RdRp) during viral replication. Due to the absence of efficient proofreading activity in their RdRps, various RNA viruses reveal high mutation frequencies, making them evolve rapidly during serial passages in cells, especially treated with a certain mutagen, like ribavirin. We have previously rescued a recombinant enhanced green fluorescence protein-tagged SRMV (rSRMV-eGFP) using reverse genetics. In this study, the rSRMV-eGFP was subjected to serial passages in ribavirin-treated cells. Due to the ribavirin-exerted selective pressure, it was speculated that viral progenies would form quasispecies after dozens of passages. Viral progenies at passage-10, -20, -30, -40, and -50 were separately subjected to next-generation sequencing (NGS), consequently revealing a total of 34 single-nucleotide variations, including five synonymous, 21 missense, and one non-sense mutations. The L sequence was found to harbor eight missense mutations during serial passaging. It was speculated that at least one high-fidelity variant was present in viral quasispecies at passage-50. If purified from the population of viral progenies, this putative variant would contribute to clarifying a molecular mechanism in viral high-fidelity replication in vitro.

Zinc finger antiviral protein (ZAP) inhibits small ruminant morbillivirus replication in vitro

Small ruminant morbillivirus (SRMV) is a highly contagious and economically important viral disease of small domestic and wild ruminants. Difficulty with its stable proliferation in ovis aries–derived cells has led to a relative lag in the study of its natural immunity and pathogenesis. Here we report the antiviral properties of ZAP against SRMV, a single-stranded negative-stranded RNA virus of the genus Morbillivirus. ZAP expression was significantly induced in sheep endometrial epithelial cells following SRMV infection. ZAP inhibited SRMV replication in cells after infection, while its overexpression in Vero-SLAM cells significantly increased their resistance to SRMV replication. The ZAP protein co-localized with SRMV RNA in the cytoplasm and ZAP-responsive elements were mapped to the 5′ untranslated region of SRMV nucleocapsid, phosphoprotein, matrix, and fusion. In summary, ZAP confers resistance to SRMV infection by directly targeting viral RNA and inhibiting viral replication. Our findings further extend the ranges of viral targets of ZAP and help elucidate the mechanism of SRMV replication.

Production of small ruminant morbillivirus, rift valley fever virus and lumpy skin disease virus in CelCradle\u2122 -500A bioreactors

BackgroundAnimal vaccination is an important way to stop the spread of diseases causing immense damage to livestock and economic losses and the potential transmission to humans. Therefore effective method for vaccine production using simple and inexpensive bioprocessing solutions is very essential. Conventional culture systems currently in use, tend to be uneconomic in terms of labor and time involved. Besides, they offer a limited surface area for growth of cells. In this study, the CelCradle™-500A was evaluated as an alternative to replace conventional culture systems in use such as Cell factories for the production of viral vaccines against small ruminant morbillivirus (PPR), rift valley fever virus (RVF) and lumpy skin disease virus (LSD).ResultsTwo types of cells Vero and primary Lamb Testis cells were used to produce these viruses. The study was done in 2 phases as a) optimization of cell growth and b) virus cultivation. Vero cells could be grown to significantly higher cell densities of 3.04 × 109 using the CelCradle™-500A with a shorter doubling time as compared to 9.45 × 108 cells in Cell factories. This represents a 19 fold increase in cell numbers as compared to seeding vs only 3.7 fold in Cell factories. LT cells achieved modestly higher cell densities of 6.7 × 108 as compared to 6.3 × 108 in Cell factories. The fold change in densities for these cells was 3 fold in the CelCradle™-500A vs 2.5 fold in Cell factories. The titers in the conventional system and the bioreactor were not significantly different. However, the Cell-specific virus yield for rift valley fever virus and lumpy skin disease virus are higher (25 virions/cell for rift valley fever virus, and 21.9 virions/cell for lumpy skin disease virus versus 19.9 virions/cell for rift valley fever virus and 10 virions/cell for lumpy skin disease virus).ConclusionsThis work represents a novel study for primary lamb testis cell culture in CellCradle™-500A bioreactors. In addition, on account of the high cell densities obtained and the linear scalability the titers could be further optimized using other culture process such us perfusion.

A new molecular approach to the diagnosis of small ruminant morbillivirus with EvaGreen based assay

A new molecular approach to the diagnosis of small ruminant morbillivirus with EvaGreen based assay

A Novel Recombinant Newcastle Disease Virus Vectored DIVA Vaccine against Peste des Petits Ruminants in Goats.

Peste des petits ruminants virus (PPRV, species: small ruminant morbillivirus) is the causative agent of the eponymous notifiable disease, the peste des petits ruminants (PPR) in wild and domestic sheep and goats. Mortality rates vary between 50% and 100%, causing significant losses of estimated 1.5 to 2 billion US Dollars per year. Live-attenuated PPRV vaccine strains are used in the field for disease prevention, but the application of a more thermostable vaccine enabling differentiation between infected and vaccinated animals (DIVA) would be highly desirable to achieve the goal of global disease eradication. We generated a recombinant Newcastle disease virus (rNDV) based on the live-attenuated NDV Clone 30 that expresses the surface protein hemagglutinin (H) of PPRV strain Kurdistan/11 (rNDV_HKur). In vitro analyses confirmed transgene expression as well as virus replication in avian, caprine, and ovine cells. Two consecutive subcutaneous vaccinations of German domestic goats with rNDV_HKur prevented clinical signs and hematogenic dissemination after an intranasal challenge with virulent PPRV Kurdistan/11. Virus shedding by different routes was reduced to a similar extent as after vaccination with the live-attenuated PPRV strain Nigeria 75/1. Goats that were either not vaccinated or inoculated with parental rNDV were used as controls. In summary, we demonstrate in a proof-of-concept study that an NDV vectored vaccine can protect against PPR. Furthermore, it provides DIVA-applicability and a high thermal tolerance.

Evaluation of mucoadhesive property and the effect of Boswellia carteri gum on intranasal vaccination against small ruminant morbillivirus infection (PPR)

ABSTRACTA study was conducted to evaluate mucoadhesive property and immunomodulatory effect of phytogenic gums from Boswellia frereana, Boswellia carteri andCommiphora myrrha on intranasal Peste des petits ruminants (PPR) vaccination in goats and sheep in an ex-vivo and in-vivo situations. Plant gums were purified, dried and compressed into 500gm tablets. Modified shear stress measurement technique was used on freshly excised trachea and intestine tissues of goat to measure peak adhesion time. Forty eight animals (24 goats and 24 sheep) were divided into eight groups (of 3 goats and 3 sheep) and immunized intranasally with gum-vaccine combinations in two ratios (1:1, 1:2). Antibody against PPR virus was measured on day 14, 28, 42 and 56 post vaccination using H-based PPR bELISA. The peak adhesion time of the different gums was transient. PPR virus antibodies were detected in all immunized goats and sheep but not in unvaccinated control. The best percentage inhibition was recorded for Boswellia carteri-vaccine combination group at a ratio of 1:1. Administration of Boswellia carteri-PPR vaccine combination through intranasal or subcutaneous route, elicited similar antibody titre, implying that the intranasal route may be used as a non-invasive alternative delivery in PPR vaccination of small ruminants.

A comprehensive global perspective on phylogenomics and evolutionary dynamics of Small ruminant morbillivirus

A string of complete genome sequences of Small ruminant morbillivirus (SRMV) have been reported from different parts of the globe including Asia, Africa and the Middle East. Despite individual genome sequence-based analysis, there is a paucity of comparative genomic and evolutionary analysis to provide overarching and comprehensive evolutionary insights. Therefore, we first enriched the existing database of complete genome sequences of SRMVs with Pakistan-originated strains and then explored overall nucleotide diversity, genomic and residue characteristics, and deduced an evolutionary relationship among strains representing a diverse geographical region worldwide. The average number of pairwise nucleotide differences among the whole genomes was found to be 788.690 with a diversity in nucleotide sequences (0.04889 ± S.D. 0.00468) and haplotype variance (0.00001). The RNA-dependent-RNA polymerase (L) gene revealed phylogenetic relationship among SRMVs in a pattern similar to those of complete genome and the nucleoprotein (N) gene. Therefore, we propose another useful molecular marker that may be employed for future epidemiological investigations. Based on evolutionary analysis, the mean evolution rate for the complete genome, N, P, M, F, H and L genes of SRMV was estimated to be 9.953 × 10–4, 1.1 × 10–3, 1.23 × 10–3, 2.56 × 10–3, 2.01 × 10–3, 1.47 × 10–3 and 9.75 × 10–4 substitutions per site per year, respectively. A recombinant event was observed in a Pakistan-originated strain (KY967608) revealing Indian strains as major (98.1%, KR140086) and minor parents (99.8%, KT860064). Taken together, outcomes of the study augment our knowledge and current understanding towards ongoing phylogenomic and evolutionary dynamics for better comprehensions of SRMVs and effective disease control interventions.