Transboundary Diseases Research Articles

Multiplexed DIVA tests for rapid detection of FMDV infection/circulation in endemic countries

Foot-and-mouth disease (FMD) is an important transboundary disease affecting domestic and wild ruminants. Due to FMD outbreaks, the annual economic losses in endemic countries range from USD 6.1 billion to 200 billion. It also restricts the export of animals/animal by-products to FMD-free countries. FMD-free countries can experience a more severe economic loss due to the culling of infected animals as experienced by the UK in 2001 outbreaks. In endemic countries outbreaks occur mainly due to unrestricted animal movements. This creates a difficult situation in an endemic setting for controlling FMD spread to nearby areas. During post-vaccination surveillance, testing of serum samples using single test may not be able to substantiate complete freedom from infection. Thus, there is a requirement of more sensitive, robust, and accurate diagnostic tests to detect the FMDV infection/virus circulation in the vaccinated population with more accuracy than the available diagnostic tests. This can be achieved by using multiple antigens and setting the criteria for the positivity/negativity of the samples. Thus, this review emphasizes the comparison and the practical utility of the available diagnostic tests which detect antibodies against single antigen with those which detect antibodies against multiple antigens in single testing. It also emphasizes the utility of these tests in PCP-FMD (Progressive Control Pathway for Foot-and-Mouth Disease) going on in endemic countries.

Biosafety and Biosecurity Challenges Facing Veterinary Diagnostic Laboratories in Lower-Middle Income Countries in Southeast Asia: A Case Study of Thailand.

Introduction:Global concerns over emerging and transboundary infectious zoonotic diseases have increased disease diagnostic demands, especially in the veterinary sector. In developing or newly developed countries where the sector often works under limited capacity, biosafety and biosecurity are unlikely to be high-priority issues. A recent development program supported by the Biological Threat Reduction Program of the Defense Threat Reduction Agency funded by the US government aimed to increase biosafety and biosecurity measures of government veterinary diagnostic and research laboratories in Thailand.Objective:The purpose of this article is to identify biosafety and biosecurity challenges, opportunities, and recommendations.Methods:Eleven government laboratory centers were assessed against the Biosafety in Microbiological and Biomedical Laboratories (BMBL) biosafety level 2 (BSL-2) requirements checklist. The BMBL assessment outcomes were then combined with the outcomes of discussion sessions, and the results of pre- and post-test questionnaires conducted during biosafety assessment workshops and self-evaluation reports using the Food and Agriculture Organization Biosafety Laboratory Mapping Tool of each laboratory center were reviewed and summarized.Results:Despite established national policies on laboratory biosafety and biosecurity, major challenges included (1) harmonization and enforcement of these policies, especially at the regional level, and (2) engagement of personnel in implementations of biosafety and biosecurity measures.Conclusion:Consistent biosafety policy and allocated resources together with regular training are required to develop sustainable biosafety and biosecurity at the national level. Collaboration between regional countries, international organizations, and donors is essential for improving biosafety and biosecurity on a global scale through setting regional priorities, enacting regulatory standards, and providing technical and financial support.

MicroRNA Expression Profile in Peripheral Blood Lymphocytes of Sheep Vaccinated with Nigeria 75/1 Peste Des Petits Ruminants Virus.

Peste des petits ruminants (PPR) is one of the highly contagious transboundary viral diseases of small ruminants. Host microRNA (miRNA) expression patterns may change in response to virus infection, and it mainly works as a post-transcriptional moderator in gene expression and affects viral pathogenesis and replication. In this study, the change of miRNA expression profile in peripheral blood lymphocyte (PBMC) from sheep inoculated with PPR vaccine virus in vivo as well as primary sheep testicular (ST) cells inoculated with PPR vaccine virus in vitro were determined via deep sequencing technology. In PBMC cells, 373 and 115 differentially expressed miRNAs (DEmiRNAs) were identified 3 days and 5 days post inoculated (dpi), respectively. While, 575 DEmiRNAs were identified when comparing miRNA profiles on 5 dpi with 3 dpi. Some of the DEmiRNAs were found to change significantly via time-course during PPR vaccine virus inoculated. Similarly, in ST cells, 136 DEmiRNAs were identified at 3 dpi in comparison with mock-inoculation. A total of 12 DEmiRNAs were validated by real-time quantitative PCR (RT-qPCR). The oar-miR-150, oar-miR-370-3p and oar-miR-411b-3p were found common differentially expressed in both PPR vaccine virus-inoculated PBMC cells and ST cells. Targets prediction and functional analysis of the DEmiRNAs uncovered mainly gathering in antigen processing and presentation pathways, protein processing in endoplasmic reticulum pathways and cell adhesion molecules pathways. Our study supplies information about the DEmiRNAs in PPR vaccine virus-inoculated PBMC cells and ST cells, and provides clues for further understanding the function of miRNAs in PPR vaccine virus replication.

SEROLOGICAL AND MOLECULAR STUDIES ON BTV IN SMALL RUMINANTS AT RISK AREA IN EGYPT POST- BT OUTBREAK IN LIBYA

Bluetongue virus (BTV) is one of the most pressing trans-boundary viral diseases that cause worldwide great loss in the bovine industry either directly or indirectly worldwide. The current study was conducted to clarify the infection status of BTV at Marsa-Matruh (The West border with Libya) which is considered a high risk spot and an entrance gate for diseases in Egypt.Retrospective study on blue tongue outbreaks worldwide were carried out and the data were collected from the OIE website for pointing out the risky spots for blue tongue that may serve as a gateway of the disease to Egypt. Small ruminants in the Marsa-Matruh province were investigated in this study (𝑛 = 555; 398 sheep and 157 goats; aged between 6 ms to 3 years old) during a period from June to October 2017. Both serum and whole blood samples were collected, for detection of BTV antibodies and antigen using ELISA, while, BTV RNA was extracted from the whole blood samples for the detection of viral RNA. The results of retrospective study revealed the disease was absent in Egypt while it was reported in the border countries including Libya and in Palestinian during 2016. Moreover, serological examination demonstrated BTV specific antibodies at 54.5% and 30.5% for sheep and for goats, with an overall prevalence of 47.7 %. However, all serum samples tested negative for detection of bluetongue (BT) antigen. Further, RT-PCR confirmed the molecular findings showed that all blood samples tested negative for BT antigen. Since the BTV is capable of crossing Egypt borders through Libya, specially quarantine measures are mandatory at Marsa-Matruh province with careful animal screening for the evidence of infection/carrier status using suitable serological technique prior to importation.

Prioritization of livestock transboundary diseases in Belgium using a multicriteria decision analysis tool based on drivers of emergence.

During the past decade, livestock diseases have (re‐)emerged in areas where they had been previously eradicated or never been recorded before. Drivers (i.e. factors of (re‐)emergence) have been identified. Livestock diseases spread irrespective of borders, and therefore, reliable methods are required to help decision‐makers to identify potential threats and try stopping their (re‐)emergence. Ranking methods and multicriteria approaches are cost‐effective tools for such purpose and were applied to prioritize a list of selected diseases (N = 29 including 6 zoonoses) based on the opinion of 62 experts in accordance with 50 drivers‐related criteria. Diseases appearing in the upper ranking were porcine epidemic diarrhoea, foot‐and‐mouth disease, low pathogenic avian influenza, African horse sickness and highly pathogenic avian influenza. The tool proposed uses a multicriteria decision analysis approach to prioritize pathogens according to drivers and can be applied to other countries or diseases.

MOLECULAR EPIDEMIOLOGY AND ECONOMIC IMPACT OF GOAT POX ON SMALL HOLDER SHEEP AND GOATS FARMERS IN NORTH CENTRAL NIGERIA

Goat pox (GTP) is a major transboundary disease of sheep and goats caused by GTP virus (GTPV) leading to significant economic losses in many regions of Africa, the Middle East and Asia where the disease is present. In this study, participatory rural appraisal techniques and active disease follow-up were used to investigate suspected GTP outbreaks in Kanam Local Government Area, North Central Nigeria to study the epidemiology and determine the financial impact of the disease amongst sheep and goats farmers. Skin scabs were collected from goats with nodular pox-like lesions during field observation. All the respondents that participated in this study confirmed previous outbreaks of goat pox in their flock and only 33% of them had in the past sought veterinary services to manage the disease medically. Morbidity and mortality rates were determined to be 47% and 33%, respectively with mortality rates relatively higher in younger animals. From figures given by respondents, the annual cost of treating the disease ranged from 538 to 1,307 USD; the annual losses due to mortality was estimated to be 2,730 USD and the annual financial losses estimated to be 6,455USD. GTPV was detected in twenty percent (2/10) of the samples analyzed using Real time polymerase chain reaction, confirming GTP in the study area. Findings of this study reaffirm that participatory rural appraisal is a reliable investigative tool to determine incidence of livestock diseases.

Introduction: Improved aquatic animal health management is vital to aquaculture's role in global food security.

Global food security and nutrition depend heavily on aquaculture, the continued growth of which is crucial as the world heads towards a human population of at least 9 billion by 2050, while harvests from wild capture will, at best, stabilise at current levels. Thus, a fundamental question is: how can we sustainably increase aquatic food production? It is clear that aquatic animal diseases present a substantial threat and, consequently, aquatic animal health management has a critical role in food security. An ecosystem approach to aquaculture will mitigate impacts on ecosystem services and biodiversity, and provide the necessary resilience to future disease threats, including those exacerbated by climate change. Due to the nature of aquatic production systems, this approach must encompass fisheries and other sectors that share the same resources. Improved aquatic animal health management must be a key component of aquaculture's future. At the national level, public-private partnerships are vital in achieving objectives of common benefit. Improved disease reporting and response is critical in the control of listed and emerging diseases and can only be achieved through government, industry and stakeholder collaboration. Great potential exists to improve biosecurity from the farm to national level, but this will only be achieved through collaboration. Industry cannot develop effective biosecurity without a clear government strategy and support, specifically legislation which provides an effective framework for safe trade. Governments have a key role in creating a regulatory environment that supports effective biosecurity and is attractive to investment; such as one that supports the development and regulatory approval of therapeutics. The improved control of transboundary diseases requires the wider and more consistent implementation of OIE standards, particularly on disease notification. This can only be achieved through improved collaboration between trading partners and by supporting low- and middle-income countries to strengthen their aquatic animal health services. There is incredible potential for aquaculture to continue its rapid growth and increase its contribution to global food security. However, sustainable growth of aquaculture is threatened by both known diseases, which we cannot effectively control, and new diseases, which may become pandemic. Recent pandemics have shown that global production systems are epidemiologically connected and, consequently, aquatic animal diseases present a shared global threat that demands global solidarity. The world now depends on a sustainable future for aquaculture and improved aquatic animal health management is critical to its continued and growing contribution to global food security.

Clinical investigation of PPR outbreak and sero-prevalence of PPR viral antibody in different areas of Bangladesh

Peste des petits ruminants (PPR) is an acute, highly contagious, world organization for animal health (OIE) notifiable and economically important transboundary viral disease of sheep and goats associated with high morbidity and mortality and caused by PPR virus. This research work was done in 2016-2017 by executing, surveillance and clinical investigation studies to determine present status of circulating PPR virus and its detection of antibody level of PPRV in different areas of Bangladesh. cELISA was conducted to detect the PPR antibody and RT-PCR also used for identification of N gene PPRV. The clinical outbreak of PPR, the total 124 samples was collected at the six locations of the country and highest case fatality (morbidity) was recorded at Jhenaidah 93.75% (75 out of 80).The highest morbidity rate and mortality rate was 69.23% and 13.07% respectively. The result of RT-PCR indicates the PPR virus circulating in the different regions of Bangladesh. For sero-prevalence of PPR antibodies of 366 serum samples were collected at different region of Bangladesh such as Chuadanga 47.81% (22 out of 47), Sirajganj 34.21% (13 out of 38), Thakurgaon 48.15% (26 out of 54), Satkhira 56.92% (37out of 65), Jhenaidah 33.33% (28 out of 84) and Chattogram 30.79 % (24 out of 78). It is reflected that the selected areas are highly suspected PPR and need to proper vaccination against PPR vaccine that can protect PPR disease in goat and sheep which helps to meet global PPR control strategy as well as contribute to achieve the two (2) number of Sustainable Development Goals (SDGs).
 Asian Australas. J. Biosci. Biotechnol. 2019, 4 (2), 109-115

Expression and characterization of the non-structural protein V of small ruminant morbillivirus.

Peste-des-petits ruminants is a transboundary viral disease of small ruminants caused by small ruminant morbillivirus (SRMV). In the present study, the full-length V gene of SRMV was constructed through site-directed mutagenesis from the P gene transcripts of the vaccine virus (Sungri/96 India) and expressed in a prokaryotic expression system. In animals, the seroconversion against this protein occurs from 14-days and is getting produced from 48h in cell culture. An indirect ELISA developed using this protein has a relative sensitivity and relative specificity of 77.73% and 73.775%, respectively as compared to c-ELISA. In this ELISA, it was observed that most of the convalescent animals elicited higher level of antibodies than vaccinated animals.

Seroprevalence and risk factors for lumpy skin disease virus seropositivity in cattle in Uganda

BackgroundLumpy skin disease (LSD) is a transboundary cattle disease caused by a Capripoxvirus of the family Poxviridae. In Uganda, documented information on the epidemiology of the disease is rare and there is no nationwide control plan, yet LSD is endemic. This study set out to investigate the seroprevalence of lumpy skin disease and determine the risk factors for LSD seropositivity, by carrying out a cross-sectional study in 21 districts of Uganda.ResultsA total of 2,263 sera samples were collected from 65 cattle herds and an indirect ELISA was used to screen for lumpy skin disease virus (LSDV) antibodies. We used univariable and multivariable mixed effect logistic regression models to identify risk factors for LSD seropositivity. The overall animal and herd-level seroprevalences were 8.7% (95% CI: 7.0–9.3) and 72.3% (95% CI: 70.0–80.3), respectively. Animal-level seroprevalence in Central region (OR = 2.13, p = 0.05, 95% CI: 1.10–4.64) was significantly different from the Northern region (Reference) and Western region (OR = 0.84, p = 0.66, 95% CI: 0.39–1.81). Management type, sex, age, mean annual precipitation > 1000 mm, and drinking from communal water sources were statistically significant risk factors for occurrence of anti-LSDV antibodies in cattle. Breed, region, herd size, contact with buffalo and other wildlife and introduction of new cattle did not have a statistically significant association with being positive for LSDV.ConclusionWe report a high herd-level LSDV seroprevalence in Uganda with a moderate animal-level seroprevalence. Cattle with the highest risk of LSD infection in Uganda are those in fenced farms, females > 25 months old, in an area with a mean annual rainfall > 1000 mm, and drinking from a communal water source.

Peste des petits ruminants: A review

Peste-des-petits-ruminants (PPR) is a notifiable, contagious and economically important transboundary viral disease of small ruminant causing high morbidity and mortality. It belongs to negative-sense, single-stranded RNA paramyxovirus of genus Morbillivirus. PPR occurs in populations of immunologically naive sheep and goats, illness and death can be high as >90%. The virus is commonly found in developing countries, especially in tropical countries where the disease is widespread. After the eradication of the rinderpest virus, which is closely relate to PPRV of small ruminants contaminated with SRMV are diagnosed having pyrexia, oculo-nasal discharges, necrotizing and erosive stomatitis, gastroenteritis, diarrhea and broncho pneumonia, whereas, gross pathology, histological findings along with laboratory confirmation of specific virus antigen, antibodies, genome in the clinical samples through a variety of serological and molecular diagnostic tests can be useful. In addition there should be social economic surveys, disease hot spot recognition and identification of role of additional species in disease transmission. SRMV control may be achieved by confinement of contaminated animals, subtraction of theoretically tainted fomites with control at import and export of sheep with goats from contaminated areas. PPR can be controlled through mass vaccination program. In the future, the preparation of a marker vaccine with a robust companion test may assist in serosurveillance for the detection of infection in vaccinated animals to control the disease. Keywords: Control; Epidemiology; Goat; Peste des petits ruminants; Sheep; Vaccination http://dx.doi.org/10.19045/bspab.2019.80063

A spatial risk assessment model framework for incursion of exotic animal disease into the European Union Member States

Disease incursion and transmission modelling can play an important role in elucidating important pathways and dynamics of transboundary diseases. It is an important pre-requisite for preparedness and rapid response. A model framework has been developed which makes use of global datasets to predict the probability of entry of exotic animal pathogens to European Union (EU) member states (MSs) via some of the most likely routes of introduction: legal trade of livestock and meat products, illegal trade of red meat, wild animal dispersion, windborne vector dispersion and human introduction of pets. The model was designed to be applicable for a wide range of pathogens, many of which have limited data. We demonstrate its application through four case study pathogens: African swine fever, Classical swine fever, Bluetongue and classical rabies.The model results highlight the differences in probability between EU MSs; the absolute values for entry via a given route differed across MSs whilst different pathogens were predicted as having the highest probability of entry for the same route across MSs. Scenario analyses suggested that the probability of entry was heavily influenced by the pathogen prevalence in the country of origin and the extent to which EU MSs pose a risk to each other; the greatest risk was predominantly from countries within the EU. While we believe the input data are obtained from high quality sources, there are still big issues with regards uncertainty in some areas, in particular with regards to prevalence of pathogens in vector populations and consistency of reporting of pathogen prevalence in animals across all countries of the world. Thus, it is inevitable that there is a high degree of uncertainty associated with the absolute values. However, the main strength of the model is the broad range of analyses over pathogens, EU MSs and routes of entry. The model is also relatively easy to update with new data and a web based visualisation tool has been developed which allows users to interrogate the results of the model. As such, we believe that the model proposed here can be a useful quantitative complement to current qualitative early warning systems, helping to drive risk-based surveillance activities, by providing detailed quantitative comparisons to indicate which pathogens are most likely to enter the EU, by which route and into which areas within Europe.

RISK OF INTRODUCING HIGHLY DANGEROUS ANIMAL VESICULAR DISEASES INTO THE RUSSIAN FEDERATION

Animal vesicular diseases are rather widespread in the world. Transboundary vesicular diseases are mainly registered in African and Asian countries. They primarily include foot and mouth disease, sheep and goat pox, peste des petits ruminants, lumpy skin disease, swine vesicular disease, vesicular stomatitis, as well as relatively new Seneca Valley virus infection. The transboundary spread of vesicular diseases is mainly caused by legal and illegal movements of animals, animal products, feeds, live vaccines; migration of wild animals across the borders from neighboring countries and passive mechanical transmission of infection. The risk of vesicular disease agent introduction with legal import of live animals into the territory of the Russian Federation is minimal. Most likely, the causative agent can be introduced when livestock products are illegally imported from the following countries: China, Turkey, India, Mongolia, Iran, Kazakhstan, etc. Violations of foreign trade regulations pose a direct threat to the epidemic situation and economic security of Russia. There is a possibility of the introduction of vesicular agents with animal feed imports into the country since some viruses can potentially survive for a long time in some ingredients. Taking into account the fact that some neighboring countries are infected with vesicular diseases, the threat of infection introduction into Russia through migration of wild animals along the North Caucasian, Central Asian and Far Eastern routes still remains.

Molecular Characterization of African Swine Fever Viruses from Outbreaks in Peri-Urban Kampala, Uganda.

African swine fever (ASF) is an infectious transboundary disease of domestic pigs and wild swine and is currently the most serious constraint to piggery in Uganda. The causative agent of ASF is a large double-stranded linear DNA virus with a complex structure. There are twenty-four ASFV genotypes described to date; however, in Uganda, only genotypes IX and X have been previously described. Inadequate ASF outbreak investigation has contributed to the delayed establishment of effective interventions to aid the control of ASF. Continuous virus characterization enhances the understanding of ASF epidemiology in terms of viral genome variations, extent, severity, and the potential source of the viruses responsible for outbreaks. We collected samples from pigs that had died of a hemorrhagic disease indicative of ASF. DNA was extracted from all samples and screened with the OIE recommended diagnostic PCR for ASF. Partial B646L (p72), full-length E183L (p54) genes, and CVR region of the P72 gene were amplified, purified, and sequenced. Web-based BLAST and MEGA X software were used for sequence analysis. ASF was confirmed in 10 of the 15 suspected pig samples. Phylogenetic analysis confirmed circulation of genotype IX by both full-length E183 (p54) and partial B646L (p72) gene sequencing. Intragenotypic resolution of the CVR region revealed major deletions in the virus genome, in some isolates of this study. The marked reduction in the number of tetrameric tandem repeats in some isolates of this study could potentially play a role in influencing the virulence of this particular genotype IX in Uganda.

Characterising the insect-borne transmission of the cattle poxvirus lumpy skin disease virus

Lumpy skin disease (LSD) is an emerging poxviral disease of cattle caused by the Capripoxvirus lumpy skin disease virus (LSDV) which generates widespread cutaneous lesions in affected animals. LSD is recognised as a transboundary high consequence disease in Africa where it contributes to rural poverty and food insecurity. In 2015 LSDV spread to southeastern Europe and currently poses a threat to cattle in neighbouring regions. Previous research indicates that LSDV is most likely transmitted by insect vectors however details of transmission pathways are unclear. This study was designed to identify the risk of transmission of LSDV posed by different insect vectors. A bovine experimental model of LSD was established in the high containment facilities at The Pirbright Institute. Potential insect vectors (Aedes aegypti, Culex quinquefasciatus, Stomoxys calcitrans and Culicoides nubeculosus) were fed on LSDV-infected cattle then incubated for up to eight days. Cattle and insects were regularly sampled to quantify LSDV present in different tissues and vector species. This data was then used to model the dynamics of LSDV infection and transmission. All four species of vector successfully acquired LSDV from infected cattle and maintained the virus up to eight days post feeding. The outputs of this research will now be used to design more effective LSD control programmes.

Detection of The Red Sea Bream Iridovirus (RSIVD) and Quality of Frozen Mackerel (Scomber japonicus) Imported Through the Port of Tanjung Mas Semarang

The imported frozen fish and fishery products can be a carrier of transboundary fish pathogen that subsequently may cause serious threat to the natural fish inhabitants, sustainability of aquaculture, human health and fisheries business in general. One of the transboundary fish diseases is Red Sea Bream Iridovirus Disease (RSIVD) caused by RSIV. The virus has been reported occurred in many countries that has active trading of fish and fisheries product with Indonesia such as Japan Cina, Taipei, Hong Kong, Korea, Malaysia, Philippines, Singapore and Thailand. In the last four years, whole frozen mackerel (Scomber japonicus) is the most imported fisheries commodity through Tanjung Emas Port. The objective of this study were to detect the presence of RSIV using PCR method according to OIE and quality as food fish for human consumption by examining for Organoleptic test, TPC, E. coli, histamine test, and the presence of parasites Anisakis sp. according to Indonesian national standard (SNI). Fish samples (25 fishes/entries) were obtained by random sampling of 10 importing entries. Results showed that there was no RSIV detected. The results of quality testing include organoleptic test, TPC, E. coli, histamine meet the SNI standard of raw fish quality. Anisakis sp. was found with the prevalence between 12 - 100%. There is one import entry that does not meet the requirements of the minimum value of organoleptic and TPC values so that it does not meet the quality standards of raw materials.

Characterising the cell-mediated immune response to Lumpy skin disease virus

Lumpy skin disease (LSD), a high-impact disease of cattle and water buffalo, is a direct threat to the European cattle industry. The causative agent is the lumpy skin disease virus (LSDV), an enveloped dsDNA virus which belongs to the family Poxviridae. Affected cattle present multiple cutaneous nodules that are characteristic of a LSDV infection. The case fatality rate of LSD is low, however affected animals suffer substantial production losses including weight loss and reduced milk production. On a wider scale, LSD is a high consequence transboundary disease with mandated export restrictions imposed on affected countries leading to substantial economic costs. Currently, there is no cure for LSDV infected cattle. Hence, mass vaccination is an important component in minimizing the spread of LSDV. There are only a handful of commercially available live attenuated vaccines in the current market. These vaccines vary in efficacy, quality, and safety, which leaves some scope for further development. This improvement is hampered by a poor understanding of the immunology of LSDV, particularly the protective immune mechanisms. In this project, we are investigating the immune response to LSDV with a focus on characterising the cell-mediated immune response. Peripheral blood mononuclear cells from LSDV immunised/infected cattle and flow cytometry assays are used to detect the production of IFN-γ by CD4+T-helper cells and CD8+cytotoxic T cells in response to live LSDV stimulation. Our goal is to improve understanding of the immune response to LSDV in order to develop effective vaccines and control programs in the future.

Maximising data to optimise animal disease early warning systems and risk assessment tools within Europe

Timely and reliable data and information availability and sharing is essential for early warning, prevention and control of transboundary diseases. While there are a growing number of global datasets capable of providing information for use in early warning systems and risk assessment (RA) tools, there are currently time-consuming data cleansing and harmonisation activities which need to be carried out before they can be reliably used and combined. Thus, using global datasets as they stand can lead to errors in RA parameterisation and results due to inherent biases in the data, e.g. missing disease prevalence data treated as a zero may inadvertently penalise those countries which do report disease outbreaks as opposed to those countries which are affected by a pathogen but do not report outbreak data. It is therefore of great importance that data are clearly provided and easy to understand and that data providers strive for greater harmonisation of database standards.In this paper the datasets utilised in the SPARE ('Spatial risk assessment framework for assessing exotic disease incursion and spread through Europe') project are described and discussed in terms of key criteria: accessibility, availability, completeness, consistency and quality. It is evident that most databases exist as information portals and not exclusively for RA purposes. Another striking issue from this assessment is the need for enhanced data sharing specifically with regards to data on illegal seizures, arthropod vector/wildlife abundance, intra-country livestock movement and national animal disease surveillance.It is hoped that the outcomes of this work will promote discussion and exchange between data providers, including the development of standardised data exchange protocols. The transformation of datasets to a common format is a considerable challenge but recommendations could and should be made on the standardisation of datasets and reporting in order to achieve a unified approach across Europe.

Prokaryotic expression, purification and evaluation of goatpox virus ORF117 protein as a diagnostic antigen in indirect ELISA to detect goatpox.

Goatpox is an economically significant transboundary viral disease of goats that is caused by goatpox virus (GTPV). This study describes the prokaryotic expression of the GTPV ORF117 protein, a homologue of vaccinia virus A27L, and evaluation of its diagnostic potential in ELISA. The GTPV ORF117 gene was cloned into the pET32a vector to express recombinant ORF117 protein (rA27L) in E. coli BL21-CodonPlus (DE3)-RIPL. The bacterial expression of the protein was confirmed by western blot analysis using anti-GTPV polyclonal antibodies that detected rA27L, which is ~ 35 kDa in size. rA27L was affinity purified under native conditions and used to assess the antibody response in an optimized indirect ELISA. The purified antigen specifically reacted with anti-GTPV and anti-SPPV serum in ELISA. A preliminary screening of random and purposive serum samples (n = 520) from sheep and goats using this optimized ELISA gave a positivity rate of 19.4 % with a diagnostic specificity of 88.7% and diagnostic sensitivity of 98.5% when compared to the gold standard serum neutralization test. Our results suggest that the indirect ELISA based on the rA27L protein has potential for serosurveillance and seromonitoring of GTPV in goats.

First molecular and serological detection of Epizootic Hemorrhagic Disease virus in white tailed deer ( Odocoileus virginianus ) from Tamaulipas, Mexico

ABSTRACT Epizootic hemorrhagic disease viruses (EHDV) are dsRNA arboviruses transmitted by biting midges of the genus Culicoides that cause disease in domestic and wild ruminants. Epizootic hemorrhagic disease (EHD) is considered the most important infectious disease of white tailed deer (WTD) in North America, some studies in Northeast Mexico reported EHDV-seropositive WTD and EHDV-infected Culicoides vectors. The increasing population of WTD that share habitat with livestock in Northeast México highlights the importance of EHD for the livestock industry in the transboundary region with the U.S. One hundred and twenty two samples from WTD in Tamaulipas state, Mexico were tested by ELISA and RT-PCR for EHDV antibodies and nucleic acid, respectively. Twelve animals were seropositive to ELISA and eleven animals were positive by RT-PCR. This is the first report of EHDV nucleic acid detection in WTD from Mexico. It is hypothesized that applying the transboundary disease approach to interdisciplinary research will help fill knowledge gaps, which could help develop countermeasures to mitigate the threat of EHDV infection in wildlife and livestock along the U.S.-Mexico border.