Investigation of Peste des petits ruminants virus circulation in Uttarakhand, India: a step towards global eradication of PPR by 2030.

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.

Flock level socio-economic and other associated risk factors for Peste des petits ruminants (PPR) exposure in sheep and goats in Madhya Pradesh state, India.

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.

Post-Vaccination Sero-Monitoring of Peste des Petits Ruminants in Sheep and Goats in Karnataka: Progress towards PPR Eradication in India.

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.

Adoption of veterinary vaccines, determining factors, and barriers in Southwest Ethiopia: Implications for livestock health and disease management strategies.

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.

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.

[Preparation of colloidal gold test strips for the detection of antibodies to peste des petits ruminants based on monoclonal antibodies to N protein].

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.

Modelling flock heterogeneity in the transmission of peste des petits ruminants virus and its impact on the effectiveness of vaccination for eradication.

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.

Farmers' valuation and willingness to pay for vaccines to protect livestock resources against priority infectious diseases in Ghana.

INTRODUCTION: Livestock vaccination coverage rates remain low in many lower and middle income countries despite effective vaccines being commonly available. Consequently, many preventable infectious livestock diseases remain highly prevalent, causing significant animal mortalities and threatening farmers' livelihood and food security. This study sought to assess farmers' maximum willingness to pay (WTP) for contagious bovine pleuropneumonia (CBPP), and peste-des-petits-ruminants (PPR) vaccination of cattle, and sheep and goats, respectively. METHODS: Overall, 350 ruminant livestock farmers were randomly selected from three districts located in the northern, middle and southern farming belts of Ghana. We implemented a double-bounded dichotomous contingent valuation experiment, where farmers indicated their WTP for vaccinating each livestock specie(s) owned at randomly assigned price points. WTP responses were analyzed using maximum likelihood estimation, and factors influencing WTP were assessed using censored regression analysis accounting for village-level clustering. RESULTS: Mean WTP for CBPP vaccination was USD 1.43 or Ghanaian Cedi (GHC) 8.63 (95% CI: GHC 7.08-GHC 10.19) per cattle. Mean WTP for PPR vaccination was USD 1.17 or GHC 7.02 (95% CI: GHC 5.99-GHC 8.05) per sheep, and USD 1.1 or GHC 6.66 (95% CI: GHC 5.89-GHC 7.44) per goat. WTP was positively associated with resilience, limited knowledge about vaccines (assessed prior to WTP experiment), farmland size, and male gender, after adjusting for other covariates. To attain 70% vaccination coverage in Ghana, vaccination costs should be no larger than GHC 5.30 (USD 0.88) for CBPP per cattle and GHC 3.89 (USD 0.65) and GHC 3.67 (USD 0.61), respectively, for PPR vaccines per sheep and goat. CONCLUSIONS: Ruminant livestock farmers in Ghana value vaccination highly, and are, on average, willing to pay vaccination costs that exceed the prevailing market prices (GHC 6 for CBPP and GHC 5 for PPR vaccination) to protect their livestock resources. To achieve 70% coverage, only minor subsidies would likely be required. These results suggest that effective disease control in these settings should be possible with appropriate distribution strategies.

Ex-ante impact of pest des petits ruminant control on micro and macro socioeconomic indicators in Senegal: A system dynamics modelling approach.

Vaccination is considered as the main tool for the Global Control and Eradication Strategy for peste des petits ruminants (PPR), and the efficacity of the PPR-vaccine in conferring long-life immunity has been established. Despite this, previous studies asserted that vaccination can be expensive and consequently, the effectiveness of disease control may not necessarily translate to overall profit for farmers. Also, the consequences of PPR control on socioeconomic indicators like food and nutrition security at a macro-national level have not been explored thoroughly. Therefore, this study seeks to assess ex-ante the impact of PPR control strategies on farm-level profitability and the socioeconomic consequences concerning food and nutrition security at a national level in Senegal. A bi-level system dynamics model, compartmentalised into five modules consisting of integrated production-epidemiological, economics, disease control, marketing, and policy modules, was developed with the STELLA Architect software, validated, and simulated for 30 years at a weekly timestep. The model was parameterised with data from household surveys from pastoral areas in Northern Senegal and relevant existing data. Nine vaccination scenarios were examined considering different vaccination parameters (vaccination coverage, vaccine wastage, and the provision of government subsidies). The findings indicate that compared to a no-vaccination scenario, all the vaccination scenarios for both 26.5% (actual vaccination coverage) and 70% (expected vaccination coverage) resulted in statistically significant differences in the gross margin earnings and the potential per capita consumption for the supply of mutton and goat meat. At the prevailing vaccination coverage (with or without the provision of government subsidies), farm households will earn an average gross margin of $69.43 (annually) more than without vaccination, and the average per capita consumption for mutton and goat meat will increase by 1.13kg/person/year. When the vaccination coverage is increased to the prescribed threshold for PPR eradication (i.e., 70%), with or without the provision of government subsidies, the average gross margin earnings would be $72.23 annually and the per capita consumption will increase by 1.23kg/person/year compared to the baseline (without vaccination). This study's findings offer an empirical justification for a sustainable approach to PPR eradication. The information on the socioeconomic benefits of vaccination can be promoted via sensitization campaigns to stimulate farmers' uptake of the practice. This study can inform investment in PPR control.

Peste des Petits Ruminants (PPR) vaccine R&D investment: financial assessment of vaccine development and administration in India.

The present study assessed the financial viability of Peste des Petits Ruminants (PPR) vaccine Research & Development (R&D) investment in India and the Gross Technology Revenue (GTR) accrual to the different stakeholders. The Net Present Value (NPV), Internal Rate of Return (IRR) and Benefit Cost Ratio (BCR) of PPR vaccine development and administration were USD 16,326.6 million (INR 130,612 crore), USD 184,54.2 million (INR 147,633 crore) and USD 21,645.6 million (INR 173,164 crore); 162.2%, 167.6% and 169.7% and 43.3:1, 48.8:1 and 57.1:1, respectively under low, medium and high disease incidence scenarios. The estimated cumulative GTR accrued during 2001-02 to 2017-18 by the innovating public research institutions (Indian Council of Agricultural Research-Indian Veterinary Research Institute (ICAR-IVRI) and Tamil Nadu Veterinary and Animal Sciences University (TANUVAS)), private vaccine producers, public sector biologicals and government revenues in terms of taxes was USD 0.696 million (INR 5.568 crore) for ICAR-IVRI and USD 0.033 million (INR 0.26 crore) for TANUVAS; USD 5.00 million (INR 40 crore); USD 7.141 million (INR 57.1 crore) and USD 0.671 million (INR 5.36 crore), respectively. Overall, financial benefits of PPR vaccine development and administration to control PPR in India outweighs the investment in manifolds.

Access to vaccination services for priority ruminant livestock diseases in Ghana: Barriers and determinants of service utilization by farmers.

INTRODUCTION: Livestock diseases are a major constraint to agricultural productivity, frequently causing significant livelihood losses for farmers, and negatively affecting public food safety and security. Vaccines provide an effective and profitable means for controlling most infectious livestock diseases, but remain underutilized. This study sought to assess the barriers and determinants of vaccination utilization for priority livestock diseases in Ghana. METHODS: We conducted a mixed-method study involving a quantitative survey with ruminant livestock farmers (N = 350) and seven focus group discussions (FGD) involving 65 ruminant livestock farmers. The survey data were analyzed, and distribution of barriers to vaccination access described. We evaluated the determinants of vaccination utilization (any use of vaccination against contagious-bovine-pleuropneumonia (CBPP) and peste-des-petits-ruminants (PPR) in 2021) using logistic regression analyses at the 0.05 significance level. FGD transcripts were analyzed deductively. We used triangulation to achieve convergence across the different datasets and analyses. RESULTS: The farmers kept an average (median) of 5 tropical livestock units (TLUs) of ruminant livestock (IQR=2.6-12.0) that were on average 8 kilometers (IQR=1.9-12.4) away from veterinary officers (VOs). Only 16% (56/350) of herds were vaccinated against the diseases. Most farmers (274/350) had limited knowledge on vaccines against CBPP and PPR infections, 63% (222/350) perceived low risk of these diseases to their herds. About half of farmers reported experiencing outbreaks of either disease in the study year (2021). Farmers scored on average 80.5 out of 98 (IQR=74-85) on the RS-14 resilience scale. After adjusting for farmers' livestock rearing experience, herd size, sex, wealth status, distance to VOs, previous disease outbreaks, and perceived risk of the diseases, vaccination utilization was negatively associated with limited knowledge (aOR=0.19, 95%CI=0.08-0.43), and positively associated with personal exposure to outbreaks in the study year (aOR=5.26, 95%CI=2.01-13.7) and increasing resilience (aOR=1.13, 95%CI=1.07-1.19). FGDs revealed farmer misconceptions about vaccines, costs of vaccines, and timely access to vaccines from VOs as additional barriers. CONCLUSIONS: Acceptability, affordability, accessibility, and availability of vaccine services represent the main barriers to vaccines utilization by ruminant livestock farmers in Ghana. Given that limited knowledge regarding the value of vaccination and shortfalls in veterinary service supply are of central importance for both the demand and supply side, more collaboration between the different stakeholders in a transdisciplinary manner to effectively address the low vaccination utilization problem is needed.

Development of cell culture based peste des petits ruminants (PPR) virus vaccine candidate from Bangladeshi isolates.

This study was conducted to develop a cell culture based PPR virus vaccine candidate using recent Bangladeshi strain of peste des petits ruminant's (PPR) virus. PPR virus was isolated from field outbreaks, confirmed by RT-PCR and used as viral inoculum for serial passaging in Vero cells for adaptation and attenuation. 60th serial passage had completed and RT-PCR and real time RT-PCR were done in every 5 passages for confirmation of PPR virus in tissue culture fluid (TCF). To assess the adaptation and attenuation cytopathology, virus titration, sequencing of both F and N genes and live animal experimentation were done. Different cellular alterations produced by PPR virus in infected Vero cells including syncytia formation, development of both intranuclear and intra cytoplasmic inclusion bodies and finally cell degradation are the indications of adaptation. The virus titre was found 2.5, 3.31, 3.55, 4.44, 4.71 and 6.5 Log10 TCID50/ml at 10th, 20th, 30th, 40th, 50th and 60th passages level respectively. In F gene sequence analysis it has been observed that few nucleotide (nt) and mino acid (aa) has been substituted as the effects of serial passaging of PPR virus in Vero cells. TCF at 60th passage level was found effective to produced protective antibody (Ab) titre in live animal experimentation. It is concluded that serially passaged and Vero cells adapted PPR virus TCF could be used as a vaccine candidate for further use to develop a potent & effective vaccine against PPR diseases.

Pathogenesis of wild-type- and vaccine-based recombinant peste des petits ruminants virus (PPRV) expressing EGFP in experimentally infected domestic goats.

Peste des petits ruminants virus (PPRV) is a highly contagious morbillivirus related to measles and canine distemper virus, mostly affecting small ruminants. The corresponding PPR disease has a high clinical impact in goats and is characterized by fever, oral and nasal erosions, diarrhoea and pneumonia. In addition, massive infection of lymphoid tissues causes lymphopaenia and immune suppression. This results in increased susceptibility to secondary bacterial infections, explaining the observed high mortality in some outbreaks. We studied the pathogenesis of PPR by experimental inoculation of Dutch domestic goats with a recombinant virulent PPRV strain modified to express EGFP and compared it to an EGFP-expressing vaccine strain of PPRV. After intratracheal inoculation with virulent PPRV, animals developed fever, viraemia and leucopaenia, and shed virus from the respiratory and gastro-intestinal tracts. Macroscopic evaluation of fluorescence at the peak of infection 7 days post-inoculation (dpi) showed prominent PPRV infection of the respiratory tract, lymphoid tissues, gastro-intestinal tract, mucosae and skin. Flow cytometry of PBMCs collected over time demonstrated a cell-associated viraemia mediated by infected lymphocytes. At 14 dpi, pathognomonic zebra stripes were detected in the mucosa of the large intestine. In contrast, vaccine strain-inoculated goats remained largely macroscopically fluorescence negative and did not present clinical signs. A low-level viraemia was detected by flow cytometry, but at necropsy no histological lesions were observed. Animals from both groups seroconverted as early as 7 dpi and sera efficiently neutralized virulent PPRV in vitro. Combined, this work presents a study of the pathogenesis of wild type- and vaccine-based PPRV in its natural host. This study shows the strength of recombinant EGFP-expressing viruses in fluorescence-guided pathogenesis studies.

A recombinant capripoxvirus expressing the F protein of peste des petits ruminants virus and the P12A3C of foot-and-mouth disease virus.

BACKGROUND: Peste des petits ruminants (PPR), foot-and-mouth disease (FMD) and sheep pox and goat pox are three important infectious diseases that infect goats, sheep and other small ruminants. It is well-known that the prevention of three diseases rely mainly on their individual vaccines. However, the vaccines have a variety of different disadvantages, such as short duration of immunity, increasing the number of vaccinations, and poor thermal stability. The purpose of this study is to construct a recombinant goat pox virus (rGPV) capable of expressing the F gene of PPRV and the P12A3C gene of FMDV as a live vector vaccine. RESULTS: The IRES, FMDV P12A3C and PPRV F genes into the multi-cloning site of the universal transfer plasmid pTKfpgigp to construct a recombinant transfer plasmid pTKfpgigpFiP12A3C, and transfected GPV-infected lamb testis (LT) cells with liposomes and produced by homologous recombination Recombinant GPV (rGPV/PPRVF-FMDVP12A3C, rGPV). The rGPV was screened and purified by green florescence protein (GFP) and xanthine-guanine-phosphoribosyltransferase gene (gpt) of Escherichia coli as selective markers, and the expression of rGPV in LT cells was detected by RT-PCR and immunofluorescence techniques. The results showed that the virus strain rGPV/PPRVF-FMDVP12A3C containing FMDV P12A3C and PPRV F genes was obtained. The exogenous genes FMDV P12A3C and PPRV F contained in rGPV were normally transcribed and translated in LT cells, and the expression products could specifically react with PPRV and FMDV antiserum. Then, the rGPV was intradermally inoculated with goats, the animal experiments showed that rGPV/PPRVF-FMDVP12A3C could induce high levels of specific antibodies against GPV, PPRV and FMDV. CONCLUSIONS: The constructed rGPV induced high levels of specific antibodies against GPV, PPRV and FMDV. The study provides a reference for " one vaccine with multiple uses " of GPV live vector vaccine.

Creation of poxvirus expressing foot-and-mouth and peste des petits ruminant disease virus proteins.

OBJECTIVE: Foot-and-mouth disease (FMD) and Peste des petits ruminant disease (PPR) are acute and severe infectious diseases of sheep and are listed as animal diseases for compulsory immunization. However, there is no dual vaccine to prevent these two diseases. The Modified Vaccinia virus Ankara strain (MVA) has been widely used in the construction of recombinant live vector vaccine because of its large capacity of foreign gene, wide host range, high safety, and immunogenicity. In this study, MVA-GFP recombinant virus skeleton was used to construct dual live vector vaccines against FMD and PPR. METHODS: The recombinant plasmid pUC57-FMDV P1-2A3CPPRV FH was synthesized and transfected into MVA-GFP infected CEF cells for homologous recombination. RESULTS: The results showed that a recombinant virus without fluorescent labeling was obtained after multiple rounds of plaque screening. The recombinant virus successfully expressed the target proteins, and the empty capsid of FMDV could be observed by transmission electron microscope (TME), and the expression levels of foreign proteins (VP1 and VP3) detected by ELISA were like those detected in FMDV-infected cells. This study laid the foundation for the successful construction of a live vector vaccine against FMD and PPR. KEY POINTS: • A recombinant MVA expressing FMDVP12A3C and PRRV HF proteins • Both the FMDV and PRRV proteins inserted into the virus were expressed • The proteins expressed by the recombinant poxvirus were assembled into VLPs.

Preparation of colloidal gold test strips for the detection of antibodies to peste des petits ruminants based on monoclonal antibodies to N protein / 生物工程学报

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.

Investigation of the effect of mutual vaccination with pest des petits ruminants and polyvalent foot and mouth disease vaccines on the immune response of sheep.

Background: Pest des petits ruminants (PPRs) and foot and mouth disease (FMD) are two viral infectious diseases affecting sheep dramatically causing great economic losses. Therefore, attention should be directed toward their control, especially through the application of well-designed vaccination schedules with specific potent vaccines. Aim: Determination of the possibility of sheep vaccination with PPR and FMD vaccines in a mutual schedule. Methods: Different groups of sheep have vaccinated with live attenuated PPR vaccine and inactivated polyvalent FMD vaccine in a mutual manner (one before the other at weekly intervals or simultaneously) followed by monitoring of the induced immunity to both vaccines using serum neutralization test (SNT) and enzyme linked immune sorbent assay (ELISA). Results: SNT and ELISA revealed that there was no antagonizing effect of any vaccine on the immune response to the mutual vaccination of sheep to the other where the obtained antibody titers in single vaccinated sheep groups were similar to those in the simultaneous vaccinated group. Conclusion: Simultaneous vaccination of sheep with PPR and polyvalent FMD vaccine is of applicable benefit saving time, effort, and stress factors on the animals.

Breaking institutional barriers to enhance women's participation in and benefit from the Peste des Petits Ruminants and Newcastle Disease vaccine value chains for Sembabule district of Uganda.

This paper describes the institutional context that shapes the visibility and positioning of women along the Peste des Petits Ruminants (PPR) and Newcastle Disease (ND) vaccine value chains for Sembabule district of Uganda. It examines the institutional barriers and opportunities that affect women's empowerment derived from inclusion of women in the decision-making processes along the livestock vaccine value chain (LVVC) and that can support viable women-centered and owned enterprises, at the vaccine development, delivery, distribution and use level. Qualitative data analysis tools such as focus group discussions, focus meals, jar voices and key informant interviews were used. Using outcome mapping, a stakeholder analysis of the critical partners in the PPR and ND value chain was done involving the regulators, vaccine manufacturers, importers, distributors, agrovets, public and private veterinary service deliverers, local leaders, women groups, and farmers. The study concluded that training related to gender equality and livestock vaccines, infrastructural and technical support to the poultry and goat women and men farmers and other chain actors are inadequate in themselves to increase vaccine adoption and improve livestock productivity in Sembabule district. Strategies that promote gender-transformative collaborative efforts among the LVVC actors and build viable gender-transformed women groups and networks are critical to increase women's participation in and benefit from the livestock vaccine value chain.

Genomic characterization of peste des petits ruminants vaccine seed "45G37/35-k", Russia.

Production of peste des petits ruminants (PPR) vaccines in Russia is based on two attenuated virus strains ("45G37/35-k" and "ARRIAH") of common origin. Here, the identity of the strain PPRV/45G37/35-k was investigated using a full genome, Illumina deep sequencing approach. Phylogenomic analysis showed that PPRV/45G37/35-k belongs to the same lineage as the widely used PPRV vaccine strain Nigeria/75/1 (lineage II). However, 248 nucleotide differences separate the genomes of these vaccine strains, indicating that the PPRV vaccine strains produced in Russia are new strains not yet recognised by the World Organization for Animal Health (WOAH). Detailed information on the safety and efficacy of these vaccines should be provided to the WOAH before further national and international distribution.

Effectiveness and profitability of preventive veterinary interventions in controlling infectious diseases of ruminant livestock in sub-Saharan Africa: a scoping review.

Agriculture in general, and livestock production in particular, serve as a livelihood source for many people in sub-Saharan Africa (SSA). In many settings, lack of control of infectious diseases hampers livestock productivity, undermining the livelihood of rural populations. This scoping review sought to identify veterinary interventions previously evaluated as well as their relative effectiveness in controlling infectious livestock diseases. To be included, papers had to be written in English, German or French, and had to describe the effectiveness and/or profitability of preventive veterinary intervention(s) against anthrax, blackleg, bovine tuberculosis, brucellosis, contagious bovine pleuropneumonia, contagious caprine pleuropneumonia, foot-and-mouth disease, goat pox, lumpy skin disease, pasteurellosis, peste des petits ruminants, and/or sheep pox in any SSA country. Of the 2748 publications initially screened, 84 met our inclusion criteria and were analyzed. Most of the studies (n = 73, 87%) evaluated the effectiveness and/or profitability of vaccination, applied exclusively, applied jointly with, or compared to strategies like deworming, antimicrobial treatment, surveillance, feed supplementation, culling and dipping in reducing morbidity and/or mortality to livestock diseases. The effectiveness and/or profitability of antimicrobial treatment (n = 5), test and slaughter (n = 5), and use of lay animal health workers (n = 1) applied exclusively, were evaluated in the other studies. Vaccination was largely found to be both effective and with positive return on investment. Ineffective vaccination was mainly due to loss of vaccine potency under unfavorable field conditions like adverse weather events, cold chain failure, and mismatch of circulating pathogen strain and the vaccines in use.In summary, vaccination is the most effective and profitable means of controlling infectious livestock diseases in SSA. However, to achieve effective control of these diseases, its implementation must integrate pathogen surveillance, and optimal vaccine delivery tools, to overcome the reported field challenges.