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1.
Front Microbiol ; 15: 1427606, 2024.
Article in English | MEDLINE | ID: mdl-38966393

ABSTRACT

Peste des petits ruminants (PPR), a disease of socioeconomic importance has been a serious threat to small ruminants. The causative agent of this disease is PPR virus (PPRV) which belongs to the genus Morbillivirus. Hemagglutinin (H) is a PPRV coded transmembrane protein embedded in the viral envelope and plays a vital role in mediating the entry of virion particle into the cell. The infected host mounts an effective humoral response against H protein which is important for host to overcome the infection. In the present study, we have investigated structural, physiological and functional properties of hemagglutinin protein using various computational tools. The sequence analysis and structure prediction analysis show that hemagglutinin protein comprises of beta sheets as the predominant secondary structure, and may lack neuraminidase activity. PPRV-H consists of several important domains and motifs that form an essential scaffold which impart various critical roles to the protein. Comparative modeling predicted the protein to exist as a homo-tetramer that binds to its cognate cellular receptors. Certain amino acid substitutions identified by multiple sequence alignment were found to alter the predicted structure of the protein. PPRV-H through its predicted interaction with TLR-2 molecule may drive the expression of CD150 which could further propagate the virus into the host. Together, our study provides new insights into PPRV-H protein structure and its predicted functions.

2.
Biol Proced Online ; 26(1): 22, 2024 Jul 05.
Article in English | MEDLINE | ID: mdl-38969986

ABSTRACT

Peste des petits ruminants (PPRV), a highly contagious viral disease, causes significant economic losses concerning sheep and goats. Recently, PPR viruses (PPRVs), have adopted new hosts and lineage IV of PPRVs represents genetic diversity within the same lineage. 350 samples, including blood, swabs, and tissues from sheep/goats, were collected during the 2020-2021 disease outbreaks in Pakistan. These samples were analysed through RT-PCR and three isolates of PPRV with accession numbers, MW600920, MW600921, and MW600922, were submitted to GenBank, based on the partial N-gene sequencing. This analysis provides a better understanding of genetic characterizations and a targeted RT-PCR approach for rapid PPRV diagnosis. An IELISA test was developed using the semi-purified antigen MW600922 isolate grown in Vero cells. The PPRV isolates currently present high divergence with the Turkish strain; conversely, similarities equivalent to 99.73% were observed for isolates collected from Pakistan. The developed indirect ELISA (IELISA) test demonstrated antibody detection rates at dilutions of 1:200 for antibodies (serum) and 1:32 for antigens. In comparison to cELISA, high specificity (85.23%) and sensitivity (90.60%) rates were observed. In contrast to the virus neutralization test (VNT), IELISA was observed to be 100% specific and 82.14% sensitive in its results. Based on these results, serological surveys conducted for PPR antibodies using IELISA can be a more effective strategy on a larger scale. Furthermore, our results demonstrate a significant breakthrough in the research in terms of cost-effectiveness and storage efficiency, and the developed IELISA test is highly recommended for use in developing countries.


Peste des petits ruminants (PPRV) is a transboundary, highly contagious, and economically significant viral disease affecting small ruminants and wildlife. PPRV, a disease that only targets animals, is the focus of the Global Eradication Programme (PPRV GEP), which aims to eradicate the disease by 2030. Following the completion of the first phase of the GEP (2017­2021), Pakistan has initiated the second phase: PPRV presence and the implementation of a control strategy. Rapid and accurate laboratory diagnosis is vital to the disease's effective control and eradication. In the present study, we have improved diagnosis by reverse transcriptase polymerase chain reaction (RT-PCR), which not only can detect low viral concentrations but also contributes to the genetic analysis of lineage-IV viruses. However, the development of cost-effective indirect ELISA (iELISA) may allow for the analysis of serum samples obtained from larger populations of small ruminants.

3.
Vet J ; : 106185, 2024 Jun 20.
Article in English | MEDLINE | ID: mdl-38908779

ABSTRACT

The aim of the present study was to investigate the frequency, genetic variability, and phylogeny of the peste des petits ruminants virus (PPRV) in ovine and caprine fetuses. During 2014 and 2017, a total of 1054 embryos/fetuses were collected in Turkey. A real-time RT-PCR assay was used for the detection of the PPRV RNA. Genetic characterization and phylogenetic analysis of the PPRV field isolates were conducted by sequencing fusion (F) protein and nucleoprotein (N) gene segments. Samples were also collected from ewes (n = 83) and nanny goats (n = 3) that had aborted and whose embryos/fetuses were found to be PPRV positive. PPRV positive embryos/fetuses were also tested for the presence of Listeria monocytogenes, Campylobacter spp., Coxiella burnetii, Chlamydophila abortus, Brucella spp., akabane virus, aino virus, bluetongue virus, border disease virus, bovine viral diarrhea virus, Cache Valley virus, and Schmallenberg virus. PPRV RNA was detected in 123 (11.7%) of the 1054 embryos/fetuses, 78 of the 83 (94%) ewes and 3 (100%) nanny goats. Border disease virus RNA and Chlamydophila abortus DNA were detected in 7 and 12 PPRV positive sheep fetuses, respectively, while other bacterial and viral agents were not detected. Phylogenetically, the field isolates in this study belong to lineage IV, and compared to other strains of lineage IV considered in this study, they showed 1 and 5 new amino acid substitutions in the F and N gene sequences, respectively. The results of the study suggest that PPRV plays an important role in abortion. Therefore, PPRV needs to be taken into consideration in sheep and goats abortions.

4.
J Virol Methods ; 329: 114971, 2024 Jun 12.
Article in English | MEDLINE | ID: mdl-38876255

ABSTRACT

Peste des petis ruminants (PPR) is an acute, highly contagious fatal disease affecting both domestic and wild small ruminants, caused by Morbillivirus caprinae (also known as peste des petis ruminants virus (PPRV)). Herein, a rapid method based on recombinase aided amplification-clustered regularly interspaced short palindromic repeats-Cas12a (RAA-CRISPR Cas12a) to detect PPRV was developed. CRISPR RNAs and RAA primers for PPRV-N (nucleocapsid) and PPRV-M (matrix) fragments were designed. The reaction system was constructed following screening and optimization. Detection could be completed within in 50 minutes at 37°C. Detection of gradient dilutions of plasmids carrying of PPRV N and M gene fragments indicated a minimum limit of detection of 10 copies/µL. There were no cross-reactions with related viruses and all tested lineages of PPRV were detected successfully. The method also showed good repeatability. The detection of clinical samples (previously detected using reverse transcription polymerase chain reaction (RT-PCR)) indicated good consistency between the RAA-CRISPR Cas12a method and RT-PCR. Thus, the RAA-CRISPR Cas12a method for rapid PPRV diagnosis has strong specificity, high sensitivity, and stable repeatability. Moreover, the results can be observed visually under blue or UV light or using lateral flow strips without complex instruments.

5.
Pathogens ; 12(12)2023 Dec 07.
Article in English | MEDLINE | ID: mdl-38133306

ABSTRACT

Peste des petits ruminants (PPR) is a highly contagious viral disease of small ruminants that threatens livelihoods and food security in developing countries and, in some cases, wild ungulate species conservation. The Greater Serengeti-Mara Ecosystem (GSME) encompasses one of the major wildlife populations of PPR virus (PPRV)-susceptible species left on earth, although no clinical disease has been reported so far. This study aimed to gain further knowledge about PPRV circulation in the GSME by identifying which factors predict PPRV seropositivity in African buffalo (Syncerus caffer). Following an ecological niche modeling framework to map host-pathogen distribution, two models of PPRV exposure and buffalo habitat suitability were performed using serological data and buffalo censuses. Western Maasai Mara National Reserve and Western Serengeti National Park were identified as high-risk areas for PPRV exposure in buffalo. Variables related to wildlife-livestock interaction contributed to the higher risk of PPRV seropositivity in buffalo, providing supportive evidence that buffalo acquire the virus through contact with infected livestock. These findings can guide the design of cost-effective PPRV surveillance using buffalo as a sentinel species at the identified high-risk locations. As more intensive studies have been carried out in Eastern GSME, this study highlights the need for investigating PPRV dynamics in Western GSME.

6.
J Gen Virol ; 104(10)2023 10.
Article in English | MEDLINE | ID: mdl-37831061

ABSTRACT

Peste des petits ruminants virus (PPRV) is known to induce transient immunosuppression in infected small ruminants by modulating several cellular pathways involved in the antiviral immune response. Our study shows that the PPRV-coded non-structural proteins C and V can interact with the cellular NF-κB p65 subunit. The PPRV-C protein interacts with the transactivation domain (TAD) while PPRV-V interacts with the Rel homology domain (RHD) of the NF-κB p65 subunit. Both viral proteins can suppress the NF-κB transcriptional activity and NF-κB-mediated transcription of cellular genes. PPRV-V protein expression can significantly inhibit the nuclear translocation of NF-κB p65 upon TNF-α stimulation, whereas PPRV-C does not affect it. The NF-κB-mediated pro-inflammatory cytokine gene expression is significantly downregulated in cells expressing PPRV-C or PPRV-V protein. Our study provides evidence suggesting a role of PPRV non-structural proteins V and C in the modulation of NF-κB signalling through interaction with the NF-κB p65 subunit.


Subject(s)
Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Animals , Humans , Peste-des-petits-ruminants virus/genetics , Peste-des-Petits-Ruminants/metabolism , Cytokines/genetics , Cytokines/metabolism , NF-kappa B/genetics , NF-kappa B/metabolism , Viral Proteins/genetics , Viral Proteins/metabolism , Ruminants , Gene Expression , Goats/genetics
7.
Viruses ; 15(9)2023 08 24.
Article in English | MEDLINE | ID: mdl-37766213

ABSTRACT

Autophagy is an essential and highly conserved catabolic process in cells, which is important in the battle against intracellular pathogens. Viruses have evolved several ways to alter the host defense mechanisms. PPRV infection is known to modulate the components of a host cell's defense system, resulting in enhanced autophagy. In this study, we demonstrate that the N protein of PPRV interacts with the core components of the class III phosphatidylinositol-3-kinase (PI3K) complex-I and results in the induction of autophagy in the host cell over, thereby expressing this viral protein. Our data shows the interaction between PPRV-N protein and different core components of the autophagy pathway, i.e., VPS34, VPS15, BECN1 and ATG14L. The PPRV-N protein can specifically interact with VPS34 of the PI3K complex-I and colocalize with the proteins of PI3K complex-I in the same sub-cellular compartment, that is, in the cytoplasm. These interactions do not affect the intracellular localization of the different host proteins. The autophagy-related genes were transcriptionally modulated in PPRV-N-expressing cells. The expression of LC3B and SQSTM1/p62 was also modulated in PPRV-N-expressing cells, indicating the induction of autophagic activity. The formation of typical autophagosomes with double membranes was visualized by transmission electron microscopy in PPRV-N-expressing cells. Taken together, our findings provide evidence for the critical role of the N protein of the PPR virus in the induction of autophagy, which is likely to be mediated by PI3K complex-I of the host.


Subject(s)
Nucleocapsid Proteins , Peste-des-petits-ruminants virus , Phosphatidylinositol 3-Kinases , Autophagy , Phosphatidylinositols
8.
Microbiol Spectr ; : e0068923, 2023 Sep 01.
Article in English | MEDLINE | ID: mdl-37655907

ABSTRACT

This study reports development and optimization of a new method for the assessment and verification of the inactivation of peste des petits ruminants virus (PPRV) by chemical agents, including Triton X-100 and commercially available viral lysis buffers. Virus inactivation was confirmed by virus isolation (VI) on Vero cells following capture of the potential residual viruses from treated samples using Nanotrap magnetic virus particles (NMVPs). Since chemical agents are cytotoxic, treated PPRV samples could not be used directly for VI on Vero cell monolayers; instead, they were diluted in Eagle's Minimum Essential Medium (EMEM) to neutralize cytotoxicity and then subjected to virus capture using NMVPs. The NMVPs and the captured viruses were then clarified on a magnetic stand, reconstituted in EMEM, and inoculated onto Vero cells that were examined for cytopathic effect (CPE). No CPE was observed on cells inoculated with treated viruses captured by NMVPs; but CPE was observed on cells inoculated with untreated viruses, including those captured by NMVPs. For further verification, the supernatants of the VI cultures (treated or untreated) were subjected to RNA extraction and PPRV-specific real-time RT-PCR (RT-qPCR). The cycle threshold values were undetectable for the supernatants of VI cultures inoculated with NMVPs reconstituted from treated PPRV but detectable for the supernatants of VI cultures inoculated with untreated PPRV or the NMVPs reconstituted from untreated PPRV, indicating complete inactivation of PPRV. This new method of verification of virus inactivation using NMVPs can be applied to other high impact viruses of agricultural or public health importance. IMPORTANCE Research including diagnosis on highly contagious viruses at the molecular level such as PCR and next-generation sequencing requires complete inactivation of the virus to ensure biosafety and biosecurity so that any accidental release of the virus does not compromise the safety of the susceptible population and the environment. In this work, peste des petits ruminants virus (PPRV) was inactivated with chemical agents, and the virus inactivation was confirmed by virus isolation (VI) using Vero cells. Since the chemical agents are cytotoxic, inactivated virus (PPRV) was diluted 1:100 to neutralize cytotoxicity, and the residual viruses (if any) were captured using Nanotrap magnetic virus particles (NMVPs). The NMVPs and the captured viruses were subjected to VI. No CPE was observed, indicating complete inactivation, and the results were further supported by real-time RT-PCR. This new protocol to verify virus inactivation can be applicable to other viruses.

9.
Protein J ; 42(6): 685-697, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37421558

ABSTRACT

The lack of specific antiviral therapy and complications associated with the existing peste des petits ruminants (PPR) vaccines accentuates the search of novel antiviral blocking agents in order to curtail the PPR infection at initial level. The synthetic hemagglutinin-neuraminidase (HN) homologous peptides may compete with the natural HN protein of PPR virus for binding to signaling lymphocytic activation molecule (SLAM) receptor, consequently, may disrupt peste des petits ruminants virus (PPRV) at entry level. Therefore, insilico analysis, synthesis, purification and subsequent characterization of HN homologous peptides were conducted in this study. The HN homologous peptides were synthesized by means of solid phase chemistry and were purified by reversed-phase-high performance liquid chromatography. The mass as well as sequence of HN homologous peptides were assessed by mass spectroscopy while its secondary structure was elucidated by circular dichroism spectroscopy. The binding (interaction) efficacy of HN homologous peptides with PPRV antibodies was assessed via indirect enzyme linked immunosorbent assay, visual detection test (red wine to purple), bathochromic shift under UV-Vis spectrophotometry and lateral flow immunochromatographic strip test. The antiviral properties and cytotoxicity of these peptides were also assessed in B95a cell line with changes in cytopathic effect and titer of PPRV (Sungri/96). The presence of green fluorescein isothiocyanate over the B95a cell surface pointed towards the binding of HN homologous peptides with surface SLAM receptor. Moreover, the intact beta sheet configuration in water and lower cytotoxicity [cytotoxic concentration 50 (CC50) > 1000 µg/ml] of these peptides signifies its in vivo use. Among HN homologous peptides, the binding efficacy and antiviral properties of pep A was relatively high in comparison to pep B and Pep ppr peptides. The prerequisite concentration of HN homologous peptides (pep A = 12.5 µg/ml; pep B = 25 µg/ml; pep ppr = 25 µg/ml) to exemplify its antiviral effect was much lower than its CC50 level. Hence, this study signifies the therapeutic potential of synthetic HN homologous peptides.

10.
Viruses ; 15(6)2023 06 05.
Article in English | MEDLINE | ID: mdl-37376624

ABSTRACT

The peste des petits ruminant (PPR) virus is a transboundary virus found in small domestic ruminants that causes high morbidity and mortality in naive herds. PPR can be effectively controlled and eradicated by vaccinating small domestic ruminants with a live-attenuated peste des petits ruminant virus (PPRV) vaccine, which provides long-lasting immunity. We studied the potency and safety of a live-attenuated vaccine in goats by detecting their cellular and humoral immune responses. Six goats were subcutaneously vaccinated with a live-attenuated PPRV vaccine according to the manufacturer's instructions, and two goats were kept in contact. Following vaccination, the goats were monitored daily, and we recorded their body temperature and clinical score. Heparinized blood and serum were collected for a serological analysis, and swab samples and EDTA blood were collected to detect the PPRV genome. The safety of the used PPRV vaccine was confirmed by the absence of PPR-related clinical signs, a negative pen-side test, a low virus genome load as detected with RT-qPCR on the vaccinated goats, and the lack horizontal transmission between the in-contact goats. The strong humoral and cellular immune responses detected in the vaccinated goats showed that the live-attenuated PPRV vaccine has a strong potency in goats. Therefore, live-attenuated vaccines against PPR can be used to control and eradicate PRR.


Subject(s)
Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Vaccines, Attenuated , Animals , Goat Diseases/diagnosis , Goats , Immunity, Humoral , Peste-des-petits-ruminants virus/genetics , Vaccines, Attenuated/adverse effects
11.
Int J Biol Macromol ; 245: 125536, 2023 Aug 01.
Article in English | MEDLINE | ID: mdl-37369256

ABSTRACT

Colorectal cancer (CRC) is a common and highly malignant neoplasm, ranking as the fourth most frequent cause of cancer-related deaths worldwide. Recently, non-human oncolytic viruses such as Peste des petits ruminants virus (PPRV) are considered as a potent candidate in the viral therapy of cancer. In the current study, the apoptotic effects of matrix (M) protein of PPRV was investigated on SW480 CRC cells. The M gene was cloned into the pcDNA™3.1/Hygro(+) expression vector and transfected into the cancer cells. The cytotoxic effects of the M protein on SW480 cells were confirmed using MTT assay. Furthermore, flow cytometry results showed that the M protein induces apoptosis in 91 % of CRC cells. Interestingly, the expression of the M gene in SW480 cells led to the up-regulation of genes including Bax, p53, and Caspase-9, as well as an increase in the Bax/Bcl-2 ratio. By using bioinformatics modeling, we hypothesized that the M protein could interact with Bax factor through its BH3-like motif and could further activate the intrinsic apoptosis pathway. Ultimately, this study provided the first evidence of the pro-apoptotic activity of PPRV M protein indicating its possible development as a promising novel anti-cancer agent.


Subject(s)
Colorectal Neoplasms , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Animals , Humans , Peste-des-petits-ruminants virus/genetics , bcl-2-Associated X Protein/genetics , Apoptosis , Goats
12.
Onderstepoort J Vet Res ; 90(1): e1-e8, 2023 Feb 28.
Article in English | MEDLINE | ID: mdl-36861926

ABSTRACT

Clinical signs suggestive of peste des petits ruminants (PPR) involved herds of small ruminants, which were described elsewhere in Sudan. Peste des petits ruminants was confirmed using an Immunocapture ELISA (IC-ELISA) assay in samples of infected and dead animals in areas of outbreaks. Therefore, to update information regarding the current situation and for assessment of the serological prevalence of PPR in small ruminants mingled at Central and Western Sudan during 2018-2019, 368 sera were collected from sheep (325 sera) and goats (43 sera) with different ages and breeds. These sera included 186 sera (173 sheep and 13 goats) from White Nile State and 182 sera (152 sheep and 30 goats) from Kordofan States. Competitive ELISA demonstrated higher prevalence of PPRV antibodies of 88.9%, 90.7% and 88.6% in both sheep and goats, goats, and sheep sera, respectively. Moreover, 100%, 94.7% and 78.5% seroprevalence values were demonstrated in South Kordofan, North Kordofan and White Nile States. The higher seroprevalence values detected in sera of unvaccinated sheep and goats indicated the wide exposure of these animals to PPRV and presence of protection following PPR viral infection. The findings of the study indicated that PPR is endemic in the surveyed areas of Sudan.Contribution: The study will contribute effectively to the global eradication programme of PPR organised by the World Organization for Animal Health (WOAH, formerly OIE) and Food and Agriculture Organization (FAO). To completely eliminate PPR from Sudan by 2030, local efforts should be directed towards effectively and wholly vaccinating small ruminants using PPRV vaccine especially in routes of seasonal animal's movement and shared grazing areas.


Subject(s)
Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Sheep Diseases , Sheep , Animals , Goats , Peste-des-Petits-Ruminants/epidemiology , Prevalence , Seroepidemiologic Studies , Sudan/epidemiology , Goat Diseases/epidemiology , Sheep Diseases/epidemiology
13.
Pol J Vet Sci ; 26(1): 137-142, 2023 Mar.
Article in English | MEDLINE | ID: mdl-36961274

ABSTRACT

Health status of Polish goat population in regard to the viral diseases remained mostly unknown. In order to determine serological status of Polish goats for selected emerging ruminant viruses, 365 serum samples collected between 2017 and 2019 in 36 districts within 10 of Polish provinces, were tested. No antibodies specific to Peste de Petite Ruminants Virus (PPRSV) and capripoxviruses (CaPV) were found in any of the tested animals. Only single individual (0.27%) was seropositive to Blutongue Virus (BTV). Antibodies directed to Schmallenberg Virus (SBV) were detected in 46 goats which represented 12.6% of the tested population. No association between seropositivity to SBV and year of sampling, province of origin, gender and age was found. In conclusion, among studied viral pathogens, currently only SBV seemed to be important for epidemiological status of Polish goats.


Subject(s)
Goat Diseases , Sheep Diseases , Viruses , Animals , Sheep , Goats , Poland/epidemiology , Antibodies, Viral , Seroepidemiologic Studies , Ruminants , Sheep Diseases/epidemiology
14.
Vet Sci ; 10(1)2023 Jan 13.
Article in English | MEDLINE | ID: mdl-36669056

ABSTRACT

(1) Background: Peste des petits ruminants (PPR) is a highly contagious animal disease affecting small ruminants, leading to significant economic losses. There has been little published data on PPR virus (PPRV) infection in the United Arab Emirates (UAE); (2) Methods: four outbreaks reported in goats and Dama gazelle in 2021 were investigated using pathological and molecular testing; (3) Results: The infected animals showed symptoms of dyspnea, oculo-nasal secretions, cough, and diarrhea. Necropsy findings were almost similar in all examined animals and compliant to the classical forms of the disease. Phylogenetic analysis based on N gene and F gene partial sequences revealed a circulation of PPRV Asian lineage IV in the UAE, and these sequences clustered close to the sequences of PPRV from United Arab Emirates, Pakistan, Tajikistan and Iran; (4) Conclusions: PPRV Asian lineage IV is currently circulating in the UAE. To the best of our knowledge, this is a first study describing PPRV in domestic small ruminant in the UAE.

15.
J Virol ; 97(2): e0171222, 2023 02 28.
Article in English | MEDLINE | ID: mdl-36651745

ABSTRACT

The pathogenic mechanisms of peste des petits ruminants virus (PPRV) infection remain poorly understood, leaving peste des petits ruminants (PPR) control and eradication especially difficult. Here, we determined that PPRV nucleocapsid (N) protein triggers formation of stress granules (SGs) to benefit viral replication. A mass spectrometry-based profiling of the interactome of PPRV N protein revealed that PPRV N protein interacted with protein kinase R (PKR)-activating protein (PACT), and this interaction was confirmed in the context of PPRV infection. PACT was essential for PPRV replication. Besides, the ectopic expression of N activated the PKR/eIF2α (α subunit of eukaryotic initiation factor 2) pathway through induction of PKR phosphorylation, but it did not induce PKR phosphorylation in PACT-deficient (PACT-/-) cells. PPRV N interacted with PACT, impairing the interaction between PACT and a PKR inhibitor, transactivation response RNA-binding protein (TRBP), which subsequently enhanced the interaction between PACT and PKR and thus promoted the activation of PKR and eIF2α phosphorylation, resulting in formation of stress granules (SGs). Consistently, PPRV infection induced SG formation through activation of the PKR/eIF2α pathway, and knockdown of N impaired PPRV-induced SG formation. PPRV-induced SG formation significantly decreased in PACT-/- cells as well. The role of SG formation in PPRV replication was subsequently investigated, which showed that SG formation plays a positive role in PPRV replication. By using an RNA fluorescence in situ hybridization assay, we found that PPRV-induced SGs hid cellular mRNA rather than viral mRNA. Altogether, our data provide the first evidence that PPRV N protein plays a role in modulating the PKR/eIF2α/SG axis and promotes virus replication through targeting PACT. IMPORTANCE Stress granule (SG) formation is a conserved cellular strategy to reduce stress-related damage regulating cell survival. A mass spectrometry-based profiling of the interactome of PPRV N protein revealed that PPRV N interacted with PACT to regulate the assembly of SGs. N protein inhibited the interaction between PACT and a PKR inhibitor, TRBP, through binding to the M1 domain of PACT, which enhanced the interaction between PACT and PKR and thus promoted PKR activation and subsequent eIF2α phosphorylation as well as SG formation. The regulatory function of N protein was strikingly abrogated in PACT-/- cells. SGs induced by PPRV infection through the PKR/eIF2α pathway are PACT dependent. The loss-of-function assay indicated that PPRV-induced SGs were critical for PPRV replication. We concluded that the PPRV N protein manipulates the host PKR/eIF2α/SG axis to favor virus replication.


Subject(s)
Nucleocapsid Proteins , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , RNA-Binding Proteins , Stress Granules , Virus Replication , Animals , Humans , In Situ Hybridization, Fluorescence , Nucleocapsid Proteins/metabolism , Peste-des-Petits-Ruminants/physiopathology , Peste-des-petits-ruminants virus/physiology , Protein Kinases/metabolism , RNA-Binding Proteins/metabolism , Stress Granules/metabolism , Virus Replication/genetics
16.
BMC Vet Res ; 19(1): 11, 2023 Jan 16.
Article in English | MEDLINE | ID: mdl-36647038

ABSTRACT

BACKGROUND: Peste des petits ruminants (PPR) disease is a cross-species infectious disease that severely affects small ruminants and causes great losses to livestock industries in various countries. Distinguishing vaccine-immunized animals from naturally infected animals is an important prerequisite for the eradication of PPR. At present PPRV are classified into lineages I through IV, and only one vaccination strain, Nigeria/75/1, belongs to lineage II, but all of the epidemic strains in China at present are from lineage IV. RESULTS: To achieve this goal, we developed an SYBR Green I real-time qRT-PCR method for rapid detection and identification of PPRV lineages II and IV by analyzing different melting curve analyses. The negative amplification of other commonly circulating viruses such as orf virus, goat poxvirus, and foot-and-mouth disease virus demonstrated that primers targeting the L gene of PPRV were extremely specific. The sensitivity of the assay was assessed based on plasmid DNA and the detection limit achieved was 100 copies of PPRV lineages II and IV. CONCLUSION: Since the method has high sensitivity, specificity, and reproducibility, it will be effectively differentiated PPRV lineages II from PPRV lineages IV in PPRV infected animals.


Subject(s)
Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Animals , Peste-des-petits-ruminants virus/genetics , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Reproducibility of Results , Peste-des-Petits-Ruminants/epidemiology , Ruminants , Goats , Goat Diseases/epidemiology
17.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-976193

ABSTRACT

@#Peste des petits ruminants(PPR)is a highly contagious disease with high mortality which is caused by peste des petits ruminants virus(PPRV)infection and leads to enormous economic losses for sheep husbandry both at home and abroad. The knowledge of interactions between PPRV and host cells is the basis of understanding of the pathogenesis and the prerequisite of disease prevention and control. This paper reviews the studies on the activation of host innate immune response by PPRV infection,the immune evasion of PPRV mediated by viral protein products,and the induction of apoptosis,autophagy and inflammasome resistance to virus,which provides a reference for further discussion on the immune evasion of PPRV and the key nodes involved in the interaction between PPRV and the host,thus providing theoretical guidance for effective prevention and control of PPR transmission.

18.
Chinese Journal of Biologicals ; (12): 1014-1020, 2023.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-996576

ABSTRACT

@#Peste des petits ruminants(PPR)is an acute and highly contagious disease caused by peste des petits ruminants virus(PPRV),which mainly infects goats and sheep with high morbidity and mortality. Because of its serious pathological damage and wide spread,PPR has caused huge economic losses to the aquaculture industry and international trade in various countries,so it is particularly important to establish a rapid and accurate detection method for the prevention and control of the disease. This paper reviews the progress in research on biological detection techniques for PPRV detection,such as routine RT-PCR,routine multiplex PCR,real-time fluorence quantitative PCR(RT-qPCR),pyrosequencing,nested PCR(nPCR),loop-mediated isothermal amplification(LAMP)and recombinase polymerase amplification(RPA),so as to provide bases and ideas for scientific detection and identification of PPRV.

19.
BMC Vet Res ; 18(1): 429, 2022 Dec 09.
Article in English | MEDLINE | ID: mdl-36494681

ABSTRACT

BACKGROUND: A Peste des petits ruminant is an acute, highly contagious and economically important transboundary viral disease of small ruminants. Despite the fact that food and agriculture organization and world organization for animal health plan to eradicate the disease by 2030, some studies indicated an increasing seropositivity of PPR infection in sheep and goats in Ethiopia. A cross-sectional study was employed to estimate the seroprevalence of PPR and to assess risk factors during the study period, February to April, 2020. Following purposive selection of the study districts, simple random sampling technique was employed to select individual animal during sample collection. A total of 384 serum samples were collected from apparently healthy sheep and goats. Competitive Enzyme Linked Immunosorbent Assay was used to detect the presence of antibodies against PPR at national veterinary institute. Descriptive statistics, Pearson's chi-square (X2) and logistic regression analysis were used is this study. RESULTS: The overall animal level seroprevalence of PPR virus was found to be 60.15% (n = 231/384) and species level prevalence rate was found to be 38.18% (n = 42) in sheep and 68.98% (n = 189) in goats in the study areas. Among the associated risk factors considered; species, sex, age and herd sizes were significantly associated (P < 0.05) with the disease occurrence. Among the associated risk factors considered in this study, species, sex, age and herd size were found to be statistically associated with the seropositivity of PPR infection. CONCLUSION: The present study finding revealed that a higher seroprevalence of PPR virus infection and this confirms peste des petits ruminant virus is circulating in Afar region. Further studies should be carried out on the entire region to determine PPR seroprevalence and to develop appropriate control and eradication strategies of PPR disease.


Subject(s)
Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Sheep Diseases , Animals , Sheep , Peste-des-Petits-Ruminants/epidemiology , Goats , Seroepidemiologic Studies , Cross-Sectional Studies , Enzyme-Linked Immunosorbent Assay/veterinary , Ruminants , Risk Factors
20.
Virusdisease ; 33(4): 413-421, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36447813

ABSTRACT

The 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 < 0.01) and goats (p < 0.01). Further, this serosurvey revealed that 60% of the epi-units (n = 185) had > 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. Supplementary Information: The online version contains supplementary material available at 10.1007/s13337-022-00796-6.

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