A field study evaluating the humoral immune response in Mongolian sheep vaccinated against sheeppox virus

Sheeppox is a transboundary disease of sheep caused by infection with the capripoxvirus sheeppox virus (SPPV). Sheeppox is found in Africa, the Middle East and Asia and is characterised by fever, multifocal cutaneous raised lesions, and death, with substantial negative impact on affected flocks. Vaccination with live attenuated capripoxvirus (CPPV) strains is an effective and widely used means of controlling sheeppox outbreaks, however there are few reports of post-vaccination field surveillance studies of sheeppox. This study used a commercially available ELISA and a fluorescence-based neutralisation assay (FVNT) to examine quantitative and temporal features of the humoral response of sheep vaccinated with a live attenuated CPPV strain in Mongolia. 400 samples were tested using the ELISA, and a subset of 45 also tested with the FVNT. There was substantial agreement between the FVNT and ELISA tests. Antibodies to CPPV were detected between 40 and 262 days post vaccination. There was no significant difference between serological status (positive / negative) and sex or age, however an inverse correlation was found between the length of time since vaccination and serological status. Animals between 90 and 180 days post-vaccination were more likely to be positive than animals greater than 180 days post vaccination. This data provides temporal parameters to consider when planning sheeppox post-vaccination monitoring programmes. In summary, our results show a commercial CPPV ELISA kit is a robust and reliable assay for use in resource-restricted low and low-middle income countries for post CPPV vaccination surveillance on a regional or national level.The attached .xls file contains all raw data used in the associated publication, " A comparative serological field study evaluating the humoral immune response in Mongolian sheep vaccinated against sheeppox virus." The dataset identifies all individual sheep by a unique identification number (ID_num). Each unique ID_num is associated with relevant metadata: Province name, Sum name, herder name coded (Herded_Id), animal species, age of the animal, in years, when sampled (Age), sex of the animal "F" or "M" (Sex), date when the animal was sampled (Date_sample_collected), date when the animal was vaccinated for sheeppox according to the vaccination records (Date_vaccinated_raw data), and for samples in which the exact date of vaccination was not available and a range of potential time was given, the midpoint date within this range (Date_vaccinated_midpoint), animals for which vaccination date was not available receive an "NA" value; time from vaccination to sampled in days (Time_from_vaccinated_midpoint); %S/P values for the ELISA test conducted in the Mongolia lab (Ag_ELISA_SCVL_OD_SP) and %S/P values for the ELISA test conducted in the UK lab (Ag_ELISA_TPI_OD_SP), results from the ELISA test classified as binary variable "Positive" or "Negative" (Ag_ELISA_SCVL_bin and Ag_ELISA_TPI_bin); titres from the fluorescence-based neutralisation assay (FVNT Titre) and results from the FVNT test classified as binary variable ("Positive" or "Negative"), all animals that were not tested by FVNT receive an "NA" value in these columns. Funding provided by: Biotechnology and Biological Sciences Research CouncilCrossref Funder Registry ID: http://dx.doi.org/10.13039/501100000268Award Number: BBS/E/I/00007031Funding provided by: Biotechnology and Biological Sciences Research CouncilCrossref Funder Registry ID: http://dx.doi.org/10.13039/501100000268Award Number: BB/E/I/00007036Funding provided by: Biotechnology and Biological Sciences Research CouncilCrossref Funder Registry ID: http://dx.doi.org/10.13039/501100000268Award Number: BB/E/I/00007037Funding provided by: Biotechnology and Biological Sciences Research CouncilCrossref Funder Registry ID: http://dx.doi.org/10.13039/501100000268Award Number: BBS/E/I/00007039Funding provided by: Biotechnology and Biological Sciences Research CouncilCrossref Funder Registry ID: http://dx.doi.org/10.13039/501100000268Award Number: BB/J004324/1Funding provided by: Biotechnology and Biological Sciences Research CouncilCrossref Funder Registry ID: http://dx.doi.org/10.13039/501100000268Award Number: BBS/E/D/20002173Funding provided by: Horizon 2020Crossref Funder Registry ID: http://dx.doi.org/10.13039/501100007601Award Number: 773701Funding provided by: Food and Agriculture Organization of the United Nations*Crossref Funder Registry ID: Award Number: TCP/MON/3603Funding provided by: IdVet*Crossref Funder Registry ID: Award Number: Funding provided by: Food and Agriculture Organization of the United NationsCrossref Funder Registry ID: Award Number: TCP/MON/3603Funding provided by: IdVetCrossref Funder Registry ID:Blood samples from sheep and associated data were collected as part of the post-vaccination surveillance programme for sheeppox implemented by the Mongolian General Authority for Veterinary Services (GAVS) in 2016

TRAF2 facilitates Vaccinia virus replication by promoting rapid virus entry.

Tumor necrosis factor receptor (TNFR)-associated factor 2 (TRAF2) is a pivotal intracellular mediator of signaling pathways downstream of TNFR1 and -2 with known pro- and antiviral effects. We investigated its role in the replication of the prototype poxvirus vaccinia virus (VACV). Loss of TRAF2 expression, either through small interfering RNA treatment of HeLa cells or through genetic knockout in murine embryonic fibroblasts (MEFs), led to significant reductions in VACV growth following low-multiplicity infection. In single-cycle infections, there was delayed production of both early and late VACV proteins as well as accelerated virus-induced alterations to cell morphology, indicating that TRAF2 influences early stages of virus replication. Consistent with an early role, uncoating assays showed normal virus attachment but delayed virus entry in the absence of TRAF2. Although alterations to c-Jun N-terminal kinase (JNK) signaling were apparent in VACV-infected TRAF2(−/−) MEFs, treatment of wild-type cells with a JNK inhibitor did not affect virus entry. Instead, treatment with an inhibitor of endosomal acidification greatly reduced virus entry into TRAF2(−/−) MEFs, suggesting that VACV is reliant on the endosomal route of entry in the absence of TRAF2. Thus, TRAF2 is a proviral factor for VACV that plays a role in promoting efficient viral entry, most likely via the plasma membrane. IMPORTANCE Tumor necrosis factor receptor-associated factors (TRAFs) are key facilitators of intracellular signaling with roles in innate and adaptive immunity and stress responses. We have discovered that TRAF2 is a proviral factor in vaccinia virus replication in both HeLa cells and mouse embryonic fibroblasts and that its influence is exercised through promotion of efficient virus entry

Identification of a functional small non-coding RNA encoded by African swine fever virus

African swine fever (ASF) poses a major threat to pig populations and food security worldwide. The disease is endemic to Africa and Eastern Europe and is rapidly emerging into Asia, where it has led to the deaths of millions of pigs in the last 12 months. The development of safe and effective vaccines to protect pigs against ASF has been hindered by lack of understanding of the complex interactions between ASFV and the host cell. We focused our work on characterizing the interactions between ASFV and sncRNAs. Although comparatively modest changes to host sncRNA abundances were observed upon ASFV infection, we discovered and characterized a novel functional ASFV-encoded sncRNA. The results from this study add important insights into ASFV host-pathogen interactions. This knowledge may be exploited to develop more effective ASFV vaccines that take advantage of the sncRNA system.</jats:p

Socio-economic impact of Foot-and-Mouth Disease outbreaks and control measures:An analysis of Mongolian outbreaks in 2017

Mongolia is a large landlocked country in central Asia and has one of the highest per capita livestock ratios in the world. During 2017 reported Foot and Mouth disease (FMD) outbreaks in Mongolia increased considerably, prompting widespread disease control measures. This study estimates the socio‐economic impact of FMD and subsequent control measures on Mongolian herders. The analysis encompassed quantification of the impact on subsistence farmers’ livelihoods and food security and estimation of the national level gross losses due to reaction and expenditure during 2017. Data were collected from 112 herders across eight Provinces that reported disease. Seventy of these herders had cases of FMD, while 42 did not have FMD in their animals but were within quarantine zones. Overall, 86/112 herders reported not drinking milk for a period of time and 38/112 reduced their meat consumption. Furthermore, 55 herders (49.1%) had to borrow money to buy food, medicines and/or pay bills or bank loans. Among herders with FMD cases, the median attack rate was 31.7%, 3.8% and 0.59% in cattle, sheep and goats respectively, with important differences across Provinces. Herders with clinical cases before the winter had higher odds of reporting a reduction in their meat consumption. National level gross losses due to FMD in 2017 were estimated using government data. The estimate of gross economic loss was 18.4 billion Mongolian‐tugriks (US$7.35 million) which equates to approximately 0.65% of the Mongolian GDP. The FMD outbreaks combined with current control measures has negatively impacted herders’ livelihoods (including herders with and without cases of FMD) which is likely to reduce stakeholder advocacy. Possible strategies that could be employed to ameliorate the negative effects of the current control policy were identified. The findings and approach are relevant to other FMD endemic regions aiming to control the disease

Financial impact of sheeppox and goatpox and estimated profitability of vaccination for subsistence farmers in selected northern states of Nigeria

Sheeppox and goatpox (SGP) are important transboundary diseases, endemic in Nigeria, causing severe clinical manifestations, impacting production, and resulting in economic losses. Vaccination is an effective control measure against SGP in endemic countries but is not currently implemented in Nigeria. This study aimed to estimate SGP financial impact and assess economic viability of SGP vaccination at the herd and regional level under different scenarios in Northern Nigeria. Integrated stochastic production and economic herd models were developed for transhumance and sedentary herds. Models were run for two disease scenarios (severely and slightly affected) and with and without vaccination, with data parameterisation from literature estimates, field survey and authors’ experience. Herd-level net financial impact of the disease and its vaccination was assessed using gross margin (GM) and partial budget analyses. These were then used to assess regional financial impact of disease and profitability of a 3-year vaccination programme using a cost-benefit analysis. The regional-analysis was performed under 0 %, 50 % and 100 % government subsidy scenarios; as a standalone programme or in combination with other existing vaccination programmes; and for risk-based and non-risk-based intervention. Median SGP losses per reproductive female were £27 (90 % CI: £31-£22), and £5 (90 % CI: £7-£3), in sedentary, and £30 (90 % CI: £41-21), and £7 (90 % CI: £10-£3), in transhumance herds, for severely and slightly affected scenarios respectively. Selling animals at a reduced price, selling fewer young animals, and reduced value of affected animals remaining in the herd were the greatest contributors to farmer’s SGP costs. SGP-affected herds realised a GM reduction of up to 121 % in sedentary and 138 % in transhumance. Median estimated regional SGP cost exceeded £24 million. Herd-level median benefits of vaccination per reproductive female were £23.76 (90 % CI: £19.28-£28.61), and £4.01 (90 % CI: £2.36-£6.31), in sedentary, and £26.85 (90 % CI: £17.99-£37.02) and £7.45 (90 % CI: £3.47-£15.14) in transhumance herds, in severely and slightly affected scenarios, respectively. Median benefit: cost ratio (BCR) for severely affected herds at 50% subsidies was 6.62 (90% CI: 5.30-8.90) for sedentary, and 5.14 (90% CI: 3.31-13.81) for transhumance herds. The regional SGP vaccination standalone programme BCR: 7–27, regional SGP vaccination with existing vaccination programme BCR: 7–228 and vaccinating high-risk areas BCR: 19–439 were found to be economically viable for all subsidy levels explored. Vaccinating low-risk areas only realised benefits with 100 % of government subsidies. This study further increases understanding of SGP’s impact within Northern Nigeria and demonstrates vaccination is an economically viable control strategy at the herd-level and also regionally, depending on the strategy and government subsidy levels considered

Quantifying and modelling the acquisition and retention of lumpy skin disease virus by haematophagus insects reveals clinically but not subclinically-affected cattle are promoters of viral transmission and key targets for control of disease outbreaks

Lumpy skin disease virus (LSDV) is a vector-transmitted poxvirus that causes disease in cattle. Vector species involved in LSDV transmission and their ability to acquire and transmit the virus are poorly characterized. Using a highly representative bovine experimental model of lumpy skin disease, we fed four model vector species (Aedes aegypti, Culex quinquefasciatus, Stomoxys calcitrans, and Culicoides nubeculosus) on LSDV-inoculated cattle in order to examine their acquisition and retention of LSDV. Subclinical disease was a more common outcome than clinical disease in the inoculated cattle. Importantly, the probability of vectors acquiring LSDV from a subclinical animal (0.006) was very low compared with that from a clinical animal (0.23), meaning an insect feeding on a subclinical animal was 97% less likely to acquire LSDV than one feeding on a clinical animal. All four potential vector species studied acquired LSDV from the host at a similar rate, but Aedes aegypti and Stomoxys calcitrans retained the virus for a longer time, up to 8 days. There was no evidence of virus replication in the vector, consistent with mechanical rather than biological transmission. The parameters obtained in this study were combined with data from studies of LSDV transmission and vector life history parameters to determine the basic reproduction number of LSDV in cattle mediated by each of the model species. This reproduction number was highest for Stomoxys calcitrans (19.1), followed by C. nubeculosus (7.1) and Ae. aegypti (2.4), indicating that these three species are potentially efficient transmitters of LSDV; this information can be used to inform LSD control programs. IMPORTANCE Lumpy skin disease virus (LSDV) causes a severe systemic disease characterized by cutaneous nodules in cattle. LSDV is a rapidly emerging pathogen, having spread since 2012 into Europe and Russia and across Asia. The vector-borne nature of LSDV transmission is believed to have promoted this rapid geographic spread of the virus; however, a lack of quantitative evidence about LSDV transmission has hampered effective control of the disease during the current epidemic. Our research shows subclinical cattle play little part in virus transmission relative to clinical cattle and reveals a low probability of virus acquisition by insects at the preclinical stage. We have also calculated the reproductive number of different insect species, therefore identifying efficient transmitters of LSDV. This information is of utmost importance, as it will help to define epidemiological control measures during LSDV epidemics and of particular consequence in resource-poor regions where LSD vaccination may be less than adequate

A Loss of Function Analysis of Host Factors Influencing Vaccinia virus Replication by RNA Interference

Vaccinia virus (VACV) is a large, cytoplasmic, double-stranded DNA virus that requires complex interactions with host proteins in order to replicate. To explore these interactions a functional high throughput small interfering RNA (siRNA) screen targeting 6719 druggable cellular genes was undertaken to identify host factors (HF) influencing the replication and spread of an eGFP-tagged VACV. The experimental design incorporated a low multiplicity of infection, thereby enhancing detection of cellular proteins involved in cell-to-cell spread of VACV. The screen revealed 153 pro- and 149 anti-viral HFs that strongly influenced VACV replication. These HFs were investigated further by comparisons with transcriptional profiling data sets and HFs identified in RNAi screens of other viruses. In addition, functional and pathway analysis of the entire screen was carried out to highlight cellular mechanisms involved in VACV replication. This revealed, as anticipated, that many pro-viral HFs are involved in translation of mRNA and, unexpectedly, suggested that a range of proteins involved in cellular transcriptional processes and several DNA repair pathways possess anti-viral activity. Multiple components of the AMPK complex were found to act as pro-viral HFs, while several septins, a group of highly conserved GTP binding proteins with a role in sequestering intracellular bacteria, were identified as strong anti-viral VACV HFs. This screen has identified novel and previously unexplored roles for cellular factors in poxvirus replication. This advancement in our understanding of the VACV life cycle provides a reliable knowledge base for the improvement of poxvirus-based vaccine vectors and development of anti-viral theraputics