ABSTRACT
Peste des petits ruminants virus (PPRV) is a multi-host pathogen with sheep and goats as main hosts. To investigate the role of cattle in the epidemiology of PPR, we simulated conditions similar to East African zero-grazing husbandry practices in a series of trials with local Zebu cattle (Bos taurus indicus) co-housed with goats (Capra aegagrus hircus). Furthermore, we developed a mathematical model to assess the impact of PPRV-transmission from cattle to goats. Of the 32 cattle intranasally infected with the locally endemic lineage IV strain PPRV/Ethiopia/Habru/2014 none transmitted PPRV to 32 co-housed goats. However, these cattle or cattle co-housed with PPRV-infected goats seroconverted. The results confirm previous studies that cattle currently play a negligible role in PPRV-transmission and small ruminant vaccination is sufficient for eradication. However, the possible emergence of PPRV strains more virulent for cattle may impact eradication. Therefore, continued monitoring of PPRV circulation and evolution is recommended.
Subject(s)
Goat Diseases , Goats , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Animals , Peste-des-Petits-Ruminants/transmission , Peste-des-Petits-Ruminants/virology , Peste-des-Petits-Ruminants/epidemiology , Cattle , Peste-des-petits-ruminants virus/immunology , Peste-des-petits-ruminants virus/physiology , Goats/virology , Goat Diseases/virology , Goat Diseases/transmission , Cattle Diseases/transmission , Cattle Diseases/virology , Cattle Diseases/epidemiology , Disease Eradication/methodsABSTRACT
BACKGROUND: Contagious and economically devastating, peste des petits ruminants (PPR) is a viral disease affecting goats and sheep, causing significant losses in livestock productivity and posing a threat to food security and rural livelihoods worldwide. OBJECTIVES: This study was conducted to assess the status of goat farmer's knowledge, attitude and practice (KAP) about PPR disease at Sylhet district of Bangladesh. METHODS: A comprehensive cross-sectional survey, conducted over 11 weeks, targeted 130 goat owners. Following a pilot study with 20 farmers, a set of 17 validated questions on PPR KAP was validated. Data collection was performed through face-to-face interviews by a trained team using KOBO Toolbox, with interpretation of responses based on established thresholds for knowledge (>65%), attitude (>75%) and practice (>70%). RESULTS: Overall, 67.7% of participants demonstrated a good level of knowledge about PPR disease. Males exhibited 1.42 times higher odds of knowledge compared to females (odds ratio = 1.42). The middle age group (31-45 years) showed significantly higher knowledge levels (83.3%, p < 0.001). Within this age group, positive practice levels were also notably higher (54.8%). Those with a higher secondary education background exhibited the most positive practice levels (>80%). Participants whose additional income came from non-governmental organization employment showed a higher positive practice level (71.4%), 1.46 times higher than other income sources. CONCLUSION: Strategic interventions should prioritize female farmers, educational empowerment and collaboration with non-governmental organizations to bolster livestock health and rural livelihoods in Bangladesh as part of national PPR control strategy to fulfil the goals of Office International des Epizooties/World Organization for Animal Health (OIE/WOAH) and Food and Agriculture Organization (FAO) PPR eradication by 2030.
Subject(s)
Farmers , Goat Diseases , Goats , Health Knowledge, Attitudes, Practice , Peste-des-Petits-Ruminants , Animals , Bangladesh/epidemiology , Peste-des-Petits-Ruminants/prevention & control , Peste-des-Petits-Ruminants/epidemiology , Goat Diseases/prevention & control , Goat Diseases/epidemiology , Goat Diseases/virology , Goat Diseases/psychology , Farmers/psychology , Male , Cross-Sectional Studies , Female , Adult , Humans , Middle Aged , Animal Husbandry/methods , Surveys and Questionnaires , Young AdultABSTRACT
Here, we report the discovery of two viruses associated with a disease characterized by severe diarrhea on a large-scale goat farm in Jilin province. Electron Microscopy observations revealed two kinds of virus particles with the sizes of 150-210 nm and 20-30 nm, respectively. Detection of 276 fecal specimens from the diseased herds showed the extensive infection of peste des petits ruminants virus (63.77%, 176/276) and caprine enterovirus (76.81%, 212/276), with a co-infection rate of 57.97% (160/276). These results were partially validated with RT-PCR, where all five PPRV-positive and CEV-positive specimens yielded the expected size of fragments, respectively, while no fragments were amplified from PPRV-negative and CEV-negative specimens. Moreover, corresponding PPRV and CEV fragments were amplified in PPRV and CEV double-positive specimens. Histopathological examinations revealed severe microscopic lesions such as degeneration, necrosis, and detachment of epithelial cells in the bronchioles and intestine. An immunohistochemistry assay detected PPRV antigens in bronchioles, cartilage tissue, intestine, and lymph nodes. Simultaneously, caprine enterovirus antigens were detected in lung, kidney, and intestinal tissues from the goats infected by the peste des petits ruminants virus. These results demonstrated the co-infection of peste des petits ruminants virus with caprine enterovirus in goats, revealing the tissue tropism for these two viruses, thus laying a basis for the future diagnosis, prevention, and epidemiological survey for these two virus infections.
Subject(s)
Coinfection , Diarrhea , Enterovirus Infections , Goat Diseases , Goats , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Animals , Peste-des-Petits-Ruminants/virology , Peste-des-Petits-Ruminants/epidemiology , Peste-des-Petits-Ruminants/pathology , Peste-des-petits-ruminants virus/isolation & purification , Peste-des-petits-ruminants virus/genetics , Goat Diseases/virology , Goat Diseases/epidemiology , China/epidemiology , Coinfection/veterinary , Coinfection/virology , Coinfection/epidemiology , Enterovirus Infections/veterinary , Enterovirus Infections/virology , Enterovirus Infections/epidemiology , Diarrhea/virology , Diarrhea/veterinary , Diarrhea/epidemiology , Enterovirus/isolation & purification , Enterovirus/genetics , Enterovirus/classification , Feces/virology , PhylogenyABSTRACT
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 50â¯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.
Subject(s)
CRISPR-Cas Systems , Goats , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Sensitivity and Specificity , Peste-des-petits-ruminants virus/genetics , Peste-des-petits-ruminants virus/isolation & purification , Animals , Peste-des-Petits-Ruminants/diagnosis , Peste-des-Petits-Ruminants/virology , Nucleocapsid Proteins/genetics , Viral Matrix Proteins/genetics , Goat Diseases/diagnosis , Goat Diseases/virology , Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , RNA, Viral/genetics , Sheep , Bacterial Proteins , Endodeoxyribonucleases , CRISPR-Associated ProteinsABSTRACT
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 (11.7â¯%) of the 1054 embryos/fetuses, 78 of the 83 (94â¯%) ewes and 3 (100â¯%) 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.
Subject(s)
Abortion, Veterinary , Goat Diseases , Goats , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Phylogeny , Sheep Diseases , Animals , Peste-des-petits-ruminants virus/genetics , Peste-des-petits-ruminants virus/isolation & purification , Sheep , Abortion, Veterinary/virology , Abortion, Veterinary/microbiology , Peste-des-Petits-Ruminants/virology , Female , Goat Diseases/virology , Goat Diseases/microbiology , Sheep Diseases/virology , Sheep Diseases/microbiology , Pregnancy , Turkey/epidemiology , Aborted Fetus/virology , Aborted Fetus/microbiology , RNA, Viral/geneticsABSTRACT
BACKGROUND: Peste des Petits Ruminants (PPR) is a world organization for animal health (WOAH) notifiable and economically important transboundary, highly communicable viral disease of small ruminants. PPR virus (PPRV) belongs to the genus Morbillivirus of the family Paramyxoviridae. AIM: The present cross-sectional epidemiological investigation was accomplished to estimate the apparent prevalence and identify the risk factors linked with peste des petits ruminants (PPR) in the previously neglected northern border regions of Pakistan. METHOD: A total of 1300 samples (serum = 328; swabs = 972) from 150 flocks/herds were compiled from sheep (n = 324), goats (n = 328), cattle (n = 324), and buffaloes (n = 324) during 2020-2021 and tested using ELISA for detection of viral antibody in sera or antigen in swabs. RESULTS: An overall apparent prevalence of 38.7% (504 samples) and an estimated true prevalence (calculated by the Rogan and Gladen estimator) of 41.0% (95% CI, 38.0-44 were recorded in the target regions. The highest apparent prevalence of 53.4% (85 samples) and the true prevalence of 57.0%, 95% Confidence Interval (CI) were documented in the Gilgit district and the lowest apparent prevalence of 53 (25.1%) and the true prevalence of 26.0%, 95% Confidence Interval (CI), 19.0-33.0) was reported in the Swat district. A questionnaire was designed to collect data about associated risk factors that were put into a univariable logistic regression to decrease the non-essential assumed risk dynamics with a P-value of 0.25. ArcGIS, 10.8.1 was used to design hotspot maps and MedCalc's online statistical software was used to calculate Odds Ratio (OR). Some of the risk factors significantly different (P < 0.05) in the multivariable logistic regression were flock/herd size, farming methods, nomadic animal movement, and outbreaks of PPR. The odds of large-sized flocks/herds were 1.7 (OR = 1.79; 95% Confidence Interval (CI) = 0.034-91.80%) times more likely to be positive than small-sized. The odds of transhumance and nomadic systems were 1.1 (OR = 1.15; 95% Confidence Interval (CI) = 0.022-58.64%) and 1.0 (OR = 1.02; 95% Confidence Interval (CI) = 0.020-51.97%) times more associated to be positive than sedentary and mixed farming systems, respectively. The odds of nomadic animal movement in the area was 0.7 (OR = 0.57; 95% Confidence Interval (CI) = 0.014-38.06%) times more associated to be positive than in areas where no nomadic movement was observed. In addition, the odds of an outbreak of PPR in the area were 1.0 (OR = 1.00; 95% Confidence Interval (CI) = 0.018-46.73%) times more associated to be positive than in areas where no outbreak of PPR was observed. CONCLUSIONS: It was concluded that many northern regions considered endemic for PPR, large and small ruminants are kept and reared together making numerous chances for virus transmission dynamic, so a big threats of disease spread exist in the region. The results of the present study would contribute to the global goal of controlling and eradicating PPR by 2030.
Subject(s)
Goat Diseases , Goats , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Sheep Diseases , Animals , Pakistan/epidemiology , Peste-des-Petits-Ruminants/epidemiology , Peste-des-Petits-Ruminants/virology , Risk Factors , Prevalence , Sheep , Cross-Sectional Studies , Goat Diseases/epidemiology , Goat Diseases/virology , Sheep Diseases/epidemiology , Sheep Diseases/virology , Peste-des-petits-ruminants virus/isolation & purification , Cattle , Buffaloes/virology , Cattle Diseases/epidemiology , Cattle Diseases/virology , Antibodies, Viral/bloodABSTRACT
This study was conducted to estimate the seroprevalence of Peste des petits ruminants virus (PPRV) and to determine the virus distribution in unvaccinated goats in the Pantnagar region of Uttarakhand state, India. A total of 212 serum samples from goats were collected randomly from various villages in three districts (Udhamsingh Nagar, Nainital, and Almora) of Uttarakhand. Serum samples were tested for anti-PPRV antibodies by a commercially available kit. RNA was extracted from the clinical samples and it was subjected to one-step RT-PCR, followed by virus isolation from positive samples. A total of 41 animals from various villages were found to be seropositive with a prevalence rate of 19.33%. PPR outbreaks were also reported from the Tarai region of Uttarakhand, and detection by PCR confirmed PPRV in 8 goats. Two representative swab samples were subjected to virus isolation in Vero cells and both samples showed typical cytopathic effects. The present study shows that PPRV is circulating in the Tarai region of Uttarakhand and mass vaccination for PPR must be followed in this region to increase herd immunity to a protective level. To the best of our knowledge, this is the first investigation of PPRV seroprevalence in unvaccinated goats of Uttarakhand, India.
Subject(s)
Goat Diseases , Goats , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Animals , Peste-des-Petits-Ruminants/epidemiology , Peste-des-Petits-Ruminants/prevention & control , Peste-des-Petits-Ruminants/virology , India/epidemiology , Peste-des-petits-ruminants virus/isolation & purification , Peste-des-petits-ruminants virus/immunology , Goat Diseases/epidemiology , Goat Diseases/virology , Goat Diseases/prevention & control , Seroepidemiologic StudiesABSTRACT
BACKGROUND: Peste des petits ruminants (PPR) is a contagious and fatal disease of sheep and goats. PPR virus (PPRV) infection induces endoplasmic reticulum (ER) stress-mediated unfolded protein response (UPR). The activation of UPR signaling pathways and their impact on apoptosis and virus replication remains controversial. OBJECTIVES: To investigate the role of PPRV-induced ER stress and the IRE1-XBP1 and IRE1-JNK pathways and their impact on apoptosis and virus replication. METHODS: The cell viability and virus replication were assessed by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, immunofluorescence assay, and Western blot. The expression of ER stress biomarker GRP78, IRE1, and its downstream molecules, PPRV-N protein, and apoptosis-related proteins was detected by Western blot and quantitative reverse transcription-polymerase chain reaction, respectively. 4-Phenylbutyric acid (4-PBA) and STF-083010 were respectively used to inhibit ER stress and IRE1 signaling pathway. RESULTS: The expression of GRP78, IRE1α, p-IRE1α, XBP1s, JNK, p-JNK, caspase-3, caspase-9, Bax and PPRV-N were significantly up-regulated in PPRV-infected cells, the expression of Bcl-2 was significantly down-regulated. Due to 4-PBA treatment, the expression of GRP78, p-IRE1α, XBP1s, p-JNK, caspase-3, caspase-9, Bax, and PPRV-N were significantly down-regulated, the expression of Bcl-2 was significantly up-regulated. Moreover, in PPRV-infected cells, the expression of p-IRE1α, p-JNK, Bax, and PPRV-N was significantly decreased, and the expression of Bcl-2 was increased in the presence of STF-083010. CONCLUSIONS: PPRV infection induces ER stress and IRE1 activation, resulting in apoptosis and enhancement of virus replication through IRE1-XBP1s and IRE1-JNK pathways.
Subject(s)
Butylamines , Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Sheep Diseases , Sulfonamides , Thiophenes , Sheep , Animals , MAP Kinase Signaling System , Caspase 3/metabolism , Caspase 9/metabolism , Endoplasmic Reticulum Chaperone BiP , Endoribonucleases/metabolism , bcl-2-Associated X Protein/metabolism , Protein Serine-Threonine Kinases , Goats/metabolism , Apoptosis , Endoplasmic Reticulum StressABSTRACT
To effectively control and eradicate PPR, the comprehensive understanding of risk factors associated with PPR exposure is vital. Hence, this study investigated socioeconomic and other associated risk determinants for PPR exposure at flock level in sheep and goats in a non-vaccination programme implemented Madhya Pradesh state India. A total of 410 sheep and goat flocks, comprised mostly of goats but also some mixed flocks, were surveyed during 2016 using a multistage random sampling procedure. Further, 230 blood samples were also collected from the farmers-reported PPR affected flocks and sera were tested using c-ELISA to confirm PPR exposure. The primary data on socioeconomic factors, farm management factors, health status, vaccination details and other epidemiological risk factors were collected from flock owners and descriptive statistics, chi-square analysis and logistic regression models were fitted to identify the significant risk factors for PPR incidence. The farmer's education, flock size, rearing pattern, and awareness of PPR vaccination were found to be significant pre-disposing risk factors for PPR exposure in the flocks. Hence, the control and eradication strategy need to be designed comprehensively considering the key social factors like education and vaccination awareness along with other flock level risk factors to eradicate PPR by 2030 in consonance with the global plan.
Subject(s)
Goat Diseases , Peste-des-Petits-Ruminants , Sheep Diseases , Animals , Sheep , Goats , Peste-des-Petits-Ruminants/epidemiology , Peste-des-Petits-Ruminants/prevention & control , Risk Factors , Socioeconomic Factors , India/epidemiology , Goat Diseases/epidemiology , Sheep Diseases/epidemiologyABSTRACT
The Peste des petits ruminant virus (PPRV) is a member of the Paramyxoviridae family and is classified into the genus Measles virus. PPRV predominantly infects small ruminants, leading to mortality rates of nearly 100%, which have caused significant economic losses in developing countries. Host proteins are important in virus replication, but the PPRV nucleocapsid (N) protein-host interacting partners for regulating PPRV replication remain unclear. The present study confirmed the interaction between PPRV-N and the host protein vimentin by co-immunoprecipitation and co-localization experiments. Overexpression of vimentin suppressed PPRV replication, whereas vimentin knockdown had the opposite effect. Mechanistically, N was subjected to degradation via the ubiquitin/proteasome pathway, where vimentin recruits the E3 ubiquitin ligase NEDD4L to fulfill N-ubiquitination, resulting in the degradation of the N protein. These findings suggest that the host protein vimentin and E3 ubiquitin ligase NEDD4L have an anti-PPRV effect.
Subject(s)
Nucleocapsid Proteins , Peste-des-petits-ruminants virus , Vimentin , Virus Replication , Nucleocapsid Proteins/metabolism , Nucleocapsid Proteins/genetics , Vimentin/metabolism , Vimentin/genetics , Animals , Peste-des-petits-ruminants virus/physiology , Peste-des-petits-ruminants virus/genetics , Peste-des-petits-ruminants virus/metabolism , Humans , Ubiquitination , Host-Pathogen Interactions , HEK293 Cells , Nedd4 Ubiquitin Protein Ligases/metabolism , Nedd4 Ubiquitin Protein Ligases/genetics , Cell Line , Peste-des-Petits-Ruminants/virology , Peste-des-Petits-Ruminants/metabolism , Protein BindingABSTRACT
Peste des petits ruminants (PPR) presents economic challenges in enzootic countries impacting small ruminant productivity. The state of Karnataka, India, implemented a mass vaccination campaign in alignment with the PPR-Global Eradication Programme (GEP) and the National Strategic Plan for PPR eradication. This study was conducted from January to March 2023 to assess seroconversion in post-vaccinated goats and sheep at the epidemiological unit (epi-unit) level, aligning with the World Organisation for Animal Health (WOAH) and the Food and Agriculture Organization (FAO) guidelines in the PPR Global Control and Eradication Strategy (GCES). Before vaccination, 3466 random serum samples were collected from small ruminants of three age groups (6-12 months, 1-2 years, and >2 years) across 116 epi-units, spanning 82 taluks in 28 districts. Post-vaccination sero-monitoring included 1102 serum samples collected from small ruminants of the 6-12-month age group only, across 111 epi-units covering 64 taluks in 23 districts. The PPRV antibody status was determined using an indigenous hemagglutinin (H) protein monoclonal antibody-based competitive ELISA kit. Pre-vaccination, the PPR seropositivity rates were 55%, 62%, and 66% in the age groups of 6-12 months, 1-2 years, and >2 years, respectively, with a 61% PPRV antibody prevalence across all the age groups. Notably, 41% of the epi-units exhibited antibody prevalence rates of ≥70%, indicating a substantial population immunity, possibly attributed to the previous vaccination program in the state since 2011. In contrast, only 17% of the epi-units had below 30% seroprevalence rates, emphasizing the need for intensified vaccination. Statistical analysis of the data revealed significant correlations (p < 0.05) between the presence of PPRV antibodies and host factors such as species, breed, and sex. Post-vaccination seroprevalence in the 6-12 months age group was found to be 73.4%, indicating the use of an efficacious vaccine. On the evaluation of vaccination immunity in the 6-12 months age group, it was revealed that over 69% of the epi-units achieved a response surpassing ≥70%, indicating a significant improvement from 42% of the epi-units in pre-vaccination. For active PPR eradication, a mass vaccination campaign (>95% coverage) targeting small ruminant populations aged >4 months is advocated, aiming to achieve the desired herd immunity of >80%. This study offers crucial insights into PPR baseline seroprevalence/immunity status and vaccine efficacy, guiding national strategies towards a PPR-free India and further supporting the global eradication initiative.
Subject(s)
Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Sheep Diseases , Sheep , Animals , Peste-des-Petits-Ruminants/epidemiology , Peste-des-Petits-Ruminants/prevention & control , Goats , Seroepidemiologic Studies , India/epidemiology , Goat Diseases/epidemiology , Goat Diseases/prevention & control , Sheep Diseases/epidemiology , Sheep Diseases/prevention & control , Vaccination/veterinary , Antibodies, Viral , Enzyme-Linked Immunosorbent Assay/veterinaryABSTRACT
Peste des petits ruminants is an acute and highly contagious disease caused by the Peste des petits ruminants virus (PPRV). Host proteins play a crucial role in viral replication. However, the effect of fusion (F) protein-interacting partners on PPRV infection is poorly understood. In this study, we found that the expression of goat plasminogen activator urokinase (PLAU) gradually decreased in a time- and dose-dependent manner in PPRV-infected goat alveolar macrophages (GAMs). Goat PLAU was subsequently identified using co-immunoprecipitation and confocal microscopy as an F protein binding partner. The overexpression of goat PLAU inhibited PPRV growth and replication, whereas silencing goat PLAU promoted viral growth and replication. Additionally, we confirmed that goat PLAU interacted with a virus-induced signaling adapter (VISA) to antagonize F-mediated VISA degradation, increasing the production of type I interferon. We also found that goat PLAU reduced the inhibition of PPRV replication in VISA-knockdown GAMs. Our results show that the host protein PLAU inhibits the growth and replication of PPRV by VISA-triggering RIG-I-like receptors and provides insight into the host protein that antagonizes PPRV immunosuppression.IMPORTANCEThe role of host proteins that interact with Peste des petits ruminants virus (PPRV) fusion (F) protein in PPRV replication is poorly understood. This study confirmed that goat plasminogen activator urokinase (PLAU) interacts with the PPRV F protein. We further discovered that goat PLAU inhibited PPRV replication by enhancing virus-induced signaling adapter (VISA) expression and reducing the ability of the F protein to degrade VISA. These findings offer insights into host resistance to viral invasion and suggest new strategies and directions for developing PPR vaccines.
Subject(s)
Goat Diseases , Goats , Host-Pathogen Interactions , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Urokinase-Type Plasminogen Activator , Viral Fusion Proteins , Animals , Adaptor Proteins, Signal Transducing/metabolism , DEAD Box Protein 58/metabolism , Goat Diseases/immunology , Goat Diseases/metabolism , Goat Diseases/virology , Goats/immunology , Goats/virology , Macrophages, Alveolar , Peste-des-Petits-Ruminants/immunology , Peste-des-Petits-Ruminants/metabolism , Peste-des-Petits-Ruminants/virology , Peste-des-petits-ruminants virus/growth & development , Peste-des-petits-ruminants virus/immunology , Peste-des-petits-ruminants virus/metabolism , Protein Binding , Urokinase-Type Plasminogen Activator/genetics , Urokinase-Type Plasminogen Activator/metabolism , Viral Fusion Proteins/metabolismABSTRACT
In Ethiopia, the use of veterinary vaccines to control animal diseases is an effective strategy. A study conducted in Southwest Ethiopia from October 2020 to October 2021 aimed to determine the adoption level of veterinary vaccines and factors affecting their use. The study used multistage random sampling to select districts and interviewed 476 farmers who had either adopted or not adopted the vaccines. The study found that certain diseases should be prioritized for vaccination to safeguard the health of cattle, sheep, goats, and poultry. These include anthrax (19.12 %), blackleg (17.65 %), foot and mouth disease (10.50 %), and lumpy skin disease (8.82 %) in cattle, and pasteurellosis (18.07 %), contagious caprine pleuropneumonia (15.97 %), peste des petits ruminants (14.15 %), and Orf (13.45 %) in sheep and goats. Newcastle disease (21.85 %), infectious bursal disease (19.33 %), and coccidiosis (17.02 %) were identified as high-priority diseases for flock health. Overall, 30.7 % of farmers were adopters of veterinary vaccines, while 69.3 % were non-adopters. The study identified several factors that influence the likelihood of adopting veterinary vaccines, including breed type (OR = 9.1, p < 0.0001), production size (OR = 9.7, p < 0.0001), production type (OR = 2.7, p < 0.0001), and farm location (OR = 9.8, p = 0.001). Common barriers to vaccination included a lack of disease knowledge, high vaccine costs, limited vaccine availability, and administration difficulties. Insights from the study can guide strategies for promoting veterinary vaccine adoption in Ethiopia. Stakeholders should pay attention to these findings since vaccine use is crucial for controlling animal diseases, enhancing animal health, and preventing economic losses. Further research is needed to investigate factors affecting enhanced veterinary vaccine adoption.
Subject(s)
Animal Diseases , Cattle Diseases , Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Sheep Diseases , Viral Vaccines , Sheep , Animals , Cattle , Ethiopia/epidemiology , Livestock , Goats , Goat Diseases/epidemiology , Goat Diseases/prevention & control , Sheep Diseases/epidemiology , Sheep Diseases/prevention & control , Peste-des-Petits-Ruminants/epidemiology , Peste-des-Petits-Ruminants/prevention & control , Animal Diseases/epidemiology , Animal Diseases/prevention & controlABSTRACT
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.
Subject(s)
Bacteriophages , Camelids, New World , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Single-Domain Antibodies , Animals , Sheep , Single-Domain Antibodies/genetics , Escherichia coli/genetics , Peste-des-petits-ruminants virus/genetics , Peste-des-Petits-Ruminants/prevention & control , Antibodies , Antigens , GoatsABSTRACT
This study aimed at investigating the genetic lineages of peste des petits ruminants virus (PPRV) currently circulating in Burkina Faso. As part of PPR surveillance in 2021 and 2022, suspected outbreaks in different regions were investigated. A risk map was produced to determine high-risk areas for PPR transmission. Based on alerts, samples were obtained from three regions and all sampled localities were confirmed to fall within PPR high risk areas. We collected swab samples from the eyes, mouth, and nose of sick goats. Some tissue samples were also collected from dead animals suspected to be infected by PPRV. In total, samples from 28 goats were analysed. Virus confirmation was performed with RT-PCR amplification targeting the nucleocapsid (N) gene. Partial N gene sequencing (350 bp) was carried out using the RT-PCR products of positives samples to characterise the circulating lineages. Eleven sequences, including ten new sequences, have been obtained. Our study identified the presence of the PPRV lineage IV in the three studied regions in Burkina Faso with a genetic heterogeneity recorded for the sequences analysed. Previously published data and results of this study suggest that PPRV lineage IV seems to be replacing lineage II in Burkina Faso.
Subject(s)
Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Animals , Peste-des-petits-ruminants virus/genetics , Peste-des-Petits-Ruminants/epidemiology , Burkina Faso/epidemiology , Molecular Epidemiology , Goat Diseases/epidemiology , Phylogeny , Ruminants , GoatsABSTRACT
Peste des petits ruminants (PPR) is a highly contagious viral disease and one of the deadliest affecting wild goats, sheep, and small ruminants; however, goats are generally more sensitive. The causative agent is the Peste des Petits Ruminants virus (PPRV), which is a single-stranded RNA virus of negative polarity belonging to the Paramyxoviridae family. In February 2020, an active outbreak of PPR was reported in a herd of a transhumant farmer in the village of Gainth Pathé (department of Kounguel, Kaffrine region, Senegal). Of the ten swabs collected from the goats, eight returned a positive result through a quantitative real-time PCR. The sample that yielded the strongest signal from the quantitative real-time PCR was further analyzed with a conventional PCR amplification and direct amplicon sequencing. A phylogenetic analysis showed that the sequence of the PPR virus obtained belonged to lineage IV. These results confirm those found in the countries bordering Senegal and reinforce the hypothesis of the importance of animal mobility between these neighboring countries in the control of PPRV. In perspective, following the discovery of this lineage IV in Senegal, a study on its dispersion is underway throughout the national territory. The results that will emerge from this study, associated with detailed data on animal movements and epidemiological data, will provide appropriate and effective information to improve PPR surveillance and control strategies with a view to its eradication.
Subject(s)
Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Sheep Diseases , Sheep , Animals , Peste-des-petits-ruminants virus/genetics , Peste-des-Petits-Ruminants/epidemiology , Senegal/epidemiology , Phylogeny , Goat Diseases/epidemiology , Ruminants , Goats , Sheep Diseases/epidemiologyABSTRACT
The fusion of haemagglutinin-neuraminidase (HN) protein of peste des petits ruminant (PPR) virus with signaling lymphocyte activation molecules (SLAM) host cell receptor consequences the virus entry and multiplication inside the host cell. The use of synthetic SLAM homologous peptides (i.e., molecular decoy for HN protein of PPR virus) may check PPR infection at the preliminary stage. Hence, the predicted SLAM homologous peptides using bioinformatics tools were synthesized by solid phase chemistry with standard Merrifield's 9-fluorenylmethoxycarbonyl (Fmoc) chemistry and were purified by reverse phase high performance liquid chromatography. The secondary structures of synthesized peptides were elucidated by circular dichroism spectroscopy. The in vitro interactions of these peptides were studied through indirect Enzyme Linked Immuno Sorbent Assay (ELISA) and visual surface plasmon UV-visible spectroscopy. The SLAM homologous peptides were able to interact with the peste des petits ruminant virus (PPRV) with varying binding efficiency. The interaction of SLAM homologous peptide with the PPR virus was ascertained by the change in the plasmon color from red wine to purple during visual detection and also by bathochromic shift in absorbance spectra under UV-visible spectrophotometry. The cytotoxic and anti-PPRV effect of these peptides were also evaluated in B95a cell line using PPR virus (Sungri/96). The cytotoxic concentration 50 (CC50 ) value of each peptide was greater than 1000 µg mL-1 . The anti-PPRV efficiency of SLAM-22 was relatively high among SLAM homologous peptides, SLAM-22 at 25 µg mL-1 concentration showed a reduction of more than log10 3 virus titer by priming of B95a cell line while the use of SLAM-15 and Muco-17 at the same concentration dropped virus titer from log10 4.8 to log10 2.5 and log10 3.1 respectively. The concentration of SLAM homologous peptide (25 µg mL-1 ) to exert its anti-PPRV effect was much less than its CC50 level (>1000 µg mL-1 ). Therefore, the synthetic SLAM homologous peptides may prove to be better agents to target PPRV.
Subject(s)
Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Animals , Peste-des-petits-ruminants virus/metabolism , Peste-des-Petits-Ruminants/metabolism , Cell Line , Viral Proteins/metabolism , Peptides/pharmacology , Peptides/metabolism , GoatsABSTRACT
A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.
Subject(s)
Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Animals , Mice , Peste-des-Petits-Ruminants/diagnosis , Peste-des-Petits-Ruminants/prevention & control , Antibodies, Monoclonal , Reproducibility of Results , Antibodies, Viral , Enzyme-Linked Immunosorbent Assay , GoatsABSTRACT
Peste des petits ruminants (PPR) is a highly contagious and fatal disease of mostly domestic goats and sheep. First reported in Uganda in 2007, the extent of peste des petits ruminants virus (PPRV) exposure, geographical distribution and risk factors of its transmission and spread are not clearly understood. In this study, we used cluster random sampling methodology to select study villages from three districts representing three different production systems along Uganda's "cattle corridor". Between October and December 2022, 2520 goat and sheep serum samples were collected from 252 households with no history of PPR vaccination in the past one year. The household heads were interviewed to assess possible risk factors of PPRV transmission using a structured questionnaire. The serum samples were screened with a commercial competitive enzyme-linked immunosorbent assay (cELISA) for PPRV antibodies. The determined overall true seroprevalence of PPRV was 27.3% [95% CI: 25.4-29.1]. The seroprevalence of PPRV antibodies in different production systems was 44.1% [95% CI: 40.6-47.7], 31.7% [95% CI: 28.4-35.0] and 6.1% [95% CI: 4.4-7.9] for pastoral, agropastoral and mixed crop-livestock production systems respectively. A mixed-effects multivariable logistic regression model revealed strong statistical evidence of association between female animals and PPRV antibody seropositivity compared to males [OR= 2.45, 95% CI: 1.7-3.5, p < 0.001]. The likelihood of being PPRV antibody seropositive significantly increased with increasing small ruminant age. Animals older than 3 years were more than three times as likely to be PPRV seropositive compared to animals aged under 1 year [OR= 3.41, 95% CI: 2.39-4.85, p < 0.001]. There was no statistical evidence of association between small ruminant species and PPRV antibody seropositivity (p = 0.423). Village flocks that interacted with neighboring flocks daily during grazing (IRR = 1.59, 95% CI: 1.19-2.13) and watering around swamps (IRR = 1.59, 95% CI: 1.19-2.13) were highly correlated with increased number of PPRV seropositive animals as compared to flocks that were more restricted in grazing and watered around other water sources other than swamps. Flocks from pastoral and agropastoral production systems were more than 10 times more likely to have seropositive animals than mixed crop-livestock flocks. Targeting PPR control interventions (vaccination and livestock movement control) to pastoral and agro-pastoral small ruminant production systems that are very prone to PPR incursions is recommended to prevent PPRV spread to low-risk smallholder mixed crop-livestock production systems.
Subject(s)
Cattle Diseases , Goat Diseases , Peste-des-Petits-Ruminants , Peste-des-petits-ruminants virus , Sheep Diseases , Male , Female , Animals , Sheep , Cattle , Peste-des-Petits-Ruminants/epidemiology , Seroepidemiologic Studies , Uganda/epidemiology , Goat Diseases/epidemiology , Sheep Diseases/epidemiology , Enzyme-Linked Immunosorbent Assay/veterinary , Risk Factors , Goats , Antibodies, Viral , LivestockABSTRACT
Peste des petits ruminants (PPR) is an acute infectious disease of small ruminants targeted for global eradication by 2030. The Global Strategy for Control and Eradication (GSCE) recommends mass vaccination targeting 70% coverage of small ruminant populations in PPR-endemic regions. These small ruminant populations are diverse with heterogeneous mixing patterns that may influence PPR virus (PPRV) transmission dynamics. This paper evaluates the impact of heterogeneous mixing on (i) PPRV transmission and (ii) the likelihood of different vaccination strategies achieving PPRV elimination, including the GSCE recommended strategy. We develop models simulating heterogeneous transmission between hosts, including a metapopulation model of PPRV transmission between villages in lowland Ethiopia fitted to serological data. Our results demonstrate that although heterogeneous mixing of small ruminant populations increases the instability of PPRV transmission-increasing the chance of fadeout in the absence of intervention-a vaccination coverage of 70% may be insufficient to achieve elimination if high-risk populations are not targeted. Transmission may persist despite very high vaccination coverage (>90% small ruminants) if vaccination is biased towards more accessible but lower-risk populations such as sedentary small ruminant flocks. These results highlight the importance of characterizing small ruminant mobility patterns and identifying high-risk populations for vaccination and support a move towards targeted, risk-based vaccination programmes in the next phase of the PPRV eradication programme. Our modelling approach also illustrates a general framework for incorporating heterogeneous mixing patterns into models of directly transmitted infectious diseases where detailed contact data are limited. This study improves understanding of PPRV transmission and elimination in heterogeneous small ruminant populations and should be used to inform and optimize the design of PPRV vaccination programmes.