Genetic characterization of African swine fever virus isolates from soft ticks at the wildlife/domestic interface in Mozambique and identification of a novel genotype

African swine fever virus (ASFV ) is one of the most threatening infectious diseases of pigs. There are not sufficient data to indicate the importance of the sylvatic cycle in the spread and maintenance of the disease locally and potentially, globally. To assess the capacity to maintain ASF in the environment, we investigated the presence of soft tickreservoirs of ASFV in Gorongosa National Park (GNP ) and its surrounding villages. A total of 1,658 soft ticks were recovered from warthog burrows and pig pens at the wildlife/livestock interface of the GNP and viral DNA was confirmed by nested PCR in 19% of Ornithodoros porcinus porcinus and 15% of O. p. domesticus . However, isolation of ASFV was only achieved in approximately 50% of the PCR ‐positive samples with nineteen haemadsorbing virus isolates recovered. These were genotyped using a combination of partial sequencing of the B646L gene (p72 ) and analysis of the central variable region (CVR ) of the B602L gene. Eleven isolates were classified as belonging to genotype II and homologous to contemporary isolates from southern Africa, the Indian Ocean and eastern Europe. Three isolates grouped within genotype V and were similar to previous isolates from Mozambique and Malawi. The remaining five isolates constituted a new, previously unidentified genotype, designated genotype XXIV . This work confirms for the first time that the virus currently circulating in eastern Europe is likely to have a wildlife origin, and that the large diversity of ASFV maintained in wildlife areas can act as a permanent sources of different strains for the domestic pig value chain in Mozambique and beyond its boundaries. Their genetic similarity to ASFV strains currently spreading across Europe justifies the need to continue studying the sylvatic cycle in this African country and other parts of southern Africa in order to identify potential hot spots of ASF emergence and target surveillance and control efforts.     

Epidemiology of African swine fever in Africa today: Sylvatic cycle versus socio‐economic imperatives

African swine fever (ASF) is believed to have evolved in eastern and southern Africa in a sylvatic cycle between common warthogs (Phacochoerus africanus) and argasid ticks of the Ornithodoros moubata complex that live in their burrows. The involvement of warthogs and possibly other wild suids in the maintenance of ASF virus means that the infection cannot be eradicated from Africa, but only prevented and controlled in domestic pig populations. Historically, outbreaks of ASF in domestic pigs in Africa were almost invariably linked to the presence of warthogs, but subsequent investigations of the disease in pigs revealed the presence of another cycle involving domestic pigs and ticks, with a third cycle becoming apparent when the disease expanded into West Africa where the sylvatic cycle is not present. The increase in ASF outbreaks that has accompanied the exponential growth of the African pig population over the last three decades has heralded a shift in the epidemiology of ASF in Africa, and the growing importance of the pig husbandry and trade in the maintenance and spread of ASF. This review, which focuses on the ASF situation between 1989 and 2017, suggests a minor role for wild suids compared with the domestic cycle, driven by socio‐economic factors that determine the ability of producers to implement the control measures needed for better management of ASF in Africa.     

Surveillance and control of African Swine Fever in free‐ranging pigs in Sardinia

African swine fever (ASF) is a notifiable infectious disease, caused by the ASF virus (ASFV), which is a DNA virus belonging to the family Asfarviridae, genus Asfivirus. This disease has gained importance in the last decade after its spread in several countries in Eastern and Central Europe, and more recently, in China. Despite the efforts made to eradicate it, ASF is still present on the Mediterranean island of Sardinia (Italy) and has been since 1978. ASF risk factors on the island have been analysed in previous studies; the role of free‐ranging pigs in virus persistence has been suggested, but has not been fully elucidated. The most recent eradication plan provides more stringent measures to combat free‐ranging pigs and any kind of illegality in the pig sector. From December 2017 to June 2018, a total of 29 depopulation actions were performed in 13 municipalities in central Sardinia, during which 2,281 free‐ranging pigs were culled and more than 50% of them were tested for ASFV and antibody presence (1,218 and 1,416, respectively). A total of 651 pigs were seropositive, with a mean seroprevalence of 53.4% (CI 95% = 50.6–56.3), and 38 were ASFV positive (virus prevalence = 2.6%; CI 95% = 2.1–3.0). To the best of our knowledge, the present study is the first to provide a complete evaluation of this millennial system of pig farming and ASFV prevalence in free‐ranging pigs. Furthermore, it has emphasised the necessity of combining the maintenance of an epidemiological surveillance program with continuous education of farmers and other people involved in pig husbandry, based on cultural and economic aspects.     

Assessment of interactions between African swine fever virus,bushpigs (Potamochoerus larvatus), Ornithodoros ticks and domestic pigs in north‐western Madagascar

Since its introduction in Madagascar in 1998, African swine fever (ASF) has severely affected national pig production and persists as a common disease in that country. Two of its natural hosts in the African continent, the bushpig (Potamochoerus larvatus) and tick vectors of the Ornithodoros moubata complex, are reported in west and central regions of the island. However, their role in the maintenance and transmission of the virus has been insufficiently studied. In this work, we tried to assess their potential role in the epidemiology of the disease in Madagascar, by assessing the levels of interaction between (i) ASF virus (ASFV) and bushpigs and (ii) between soft ticks and domestic and wild suids in north‐western Madagascar. Twenty‐seven sera and 35 tissue samples from bushpigs were collected and analysed for the presence of anti‐ASF antibodies and viral DNA. In addition, the sera from 27 bushpigs and 126 domestic pigs were analysed with an ELISA test for the detection of antibodies against salivary antigens from Ornithodoros ticks. No circulation of ASFV or anti‐ASFV antibodies nor anti‐tick antibodies were detected in bushpigs. However, seven of the domestic pig sera (5.6% of the total sample population) were antibody positive for O. moubata antigens. The probability of freedom from ASFV in the bushpig population using Bayesian statistical methods ranged between 73% and 84%. The probabilities of absence of anti‐tick antibodies in domestic and wild pigs were estimated at 63% and 71%, respectively. These preliminary results suggest that bushpigs are unlikely to play a significant role in the maintenance and transmission of ASFV in Madagascar. Nevertheless, further ASFV surveys are needed on that species to confirm this assumption. In addition, the presence of antibodies against O. moubata in domestic pigs suggests that soft ticks may be able to maintain ASFV within a domestic pig cycle in areas of Madagascar where they remain present.     

Co‐circulation of multiple genotypes of African swine fever viruses among domestic pigs in Zambia (2013–2015)

During 2013–2015, several and severe outbreaks of African swine fever (ASF ) affected domestic pigs in six provinces of Zambia. Genetic characterization of ASF viruses (ASFV s) using standardized genotyping procedures revealed that genotypes I, II and XIV were associated with these outbreaks. Molecular and epidemiological data suggest that genotype II ASFV (Georgia 2007/1‐like) detected in Northern Province of Zambia may have been introduced from neighbouring Tanzania. Also, a genotype II virus detected in Eastern Province of Zambia showed a p54 phylogenetic relationship that was inconsistent with that of p72, underscoring the genetic variability of ASFV s. While it appears genotype II viruses detected in Zambia arose from a domestic pig cycle, genotypes I and XIV possibly emerged from a sylvatic cycle. Overall, this study demonstrates the co‐circulation of multiple genotypes of ASFV s, involvement of both the sylvatic and domestic pig cycle in ASF outbreaks in Zambia and possible trans‐boundary spread of the disease in south‐eastern Africa. Indeed, while there is need for regional or international concerted efforts in the control of ASF , understanding pig marketing practices, pig population dynamics, pig housing and rearing systems and community engagement will be important considerations when designing future prevention and control strategies of this disease in Zambia.     

Understanding African Swine Fever infection dynamics in Sardinia using a spatially explicit transmission model in domestic pig farms

African swine fever virus (ASFV ) has been endemic in Sardinia since 1978, resulting in severe losses for local pig producers and creating important problems for the island's veterinary authorities. This study used a spatially explicit stochastic transmission model followed by two regression models to investigate the dynamics of ASFV spread amongst domestic pig farms, to identify geographic areas at highest risk and determine the role of different susceptible pig populations (registered domestic pigs, non‐registered domestic pigs [brado ] and wild boar) in ASF occurrence. We simulated transmission within and between farms using an adapted version of the previously described model known as Be‐FAST . Results from the model revealed a generally low diffusion of ASF in Sardinia, with only 24% of the simulations resulting in disease spread, and for each simulated outbreak on average only four farms and 66 pigs were affected. Overall, local spread (indirect transmission between farms within a 2 km radius through fomites) was the most common route of transmission, being responsible for 98.6% of secondary cases. The risk of ASF occurrence for each domestic pig farm was estimated from the spread model results and integrated in two regression models together with available data for brado and wild boar populations. There was a significant association between the density of all three populations (domestic pigs, brado , and wild boar) and ASF occurrence in Sardinia. The most significant risk factors were the high densities of brado (OR = 2.2) and wild boar (OR = 2.1). The results of both analyses demonstrated that ASF epidemiology and infection dynamics in Sardinia create a complex and multifactorial disease situation, where all susceptible populations play an important role. To stop ASF transmission in Sardinia, three main factors (improving biosecurity on domestic pig farms, eliminating brado practices and better management of wild boars) need to be addressed.     

Clinical symptoms of African swine fever in domestic pig farms in the Republic of Korea, 2019

This study describes the clinical characteristics of the African swine fever (ASF) outbreaks in 14 domestic pig farms in the Republic of Korea. ASF outbreak was identified by farmers' notifications in 11 farms and by active surveillance in the remaining three. At the time of notification, farmers reported sudden death, abortion and anorexia in sows. Death was the primary symptom identified by farmers in fattener pigs. The number of animals exhibiting clinical symptoms did not exceed four heads at notification, and the number of asymptomatic virus positives was four heads per farm on average. As ASF virus was detected only in the same pig house (in a pen for fattener pigs) in each of 14 ASF outbreak farms, there has been no evidence of house‐to‐house viral spread within any of the ASF outbreak farms. This in turn supports our hypothesis that infection was successfully detected during its initial phase.     

Monitoring of African Swine Fever in the Wild Boar Population of the Most Recent Endemic Area of Spain

Wild boars are natural hosts for African swine fever (ASF). The ASF virus (ASFV) can persist for long periods in the environment, such as in ticks and contaminated products, which may be sources of infection for wild boar populations. African swine fever was eradicated in domestic pig populations in Spain in 1995, after 35 years of significant effort. To determine whether ASFV can persist in wild boar hosts after it has been eradicated from domestic pigs and to study the role of wild boar in helping ASFV persist in the environment, we checked for the presence of ASFV in wild boars in Doñana National Park, one of the largest natural habitats of wild boar in Spain and one of the last areas where ASF was endemic prior its eradication. Samples from 158 animals collected between 2006 and 2010 were analysed using serological and nucleic acid‐based diagnostic techniques recommended by the World Organization for Animal Health (OIE). None of the samples was found to be positive. These results confirm the absence of disease in wildlife in what was once one of the areas most affected by ASF in Spain, and they suggest that wild boars play a limited role in ASFV persistence. These results confirm that ASFV cannot persist in isolated wild boar populations for long periods of time without the interaction of other factors such as re‐infection by contact with domestic pigs or by feeding on contaminated swill.     

Experimental Infection of Domestic Pigs with African Swine Fever Virus Lithuania 2014 Genotype II Field Isolate

An experimental infection was conducted to evaluate horizontal transmission, clinical, virological and humoral response induced in domestic pigs infected with African swine fever (ASF) genotype II virus circulating in 2014 into the European Union (EU). Ten naive pigs were placed in contact with eight pigs experimentally inoculated with the Lithuanian LT14/1490 ASF virus (ASFV) responsible for the first ASF case detected in wild boar in Lithuania in January 2014. Clinical examination and rectal temperature were recorded each day. Blood sampling from every animal was carried out twice weekly. Blood samples were examined for presence of ASF virus‐specific antibodies and for determining the ASFV viral load. From the obtained results, it was concluded that the Lithuanian ASFV induced an acute disease which resulted in 94, 5% mortality. The disease was easily detected by real‐time PCR prior to the onset of clinical signs and 33% of the animals seroconverted. All findings were in accordance with observations previously made in domestic pigs and wild boar when infected with ASF genotype II viruses characterized by a high virulence. One in‐contact pig remained asymptomatic and survived the infection. The role of such animals in virus transmission would need further investigation.     

Short time window for transmissibility of African swine fever virus from a contaminated environment

Since the introduction of African swine fever virus (ASFV ) into the Baltic states and Poland in 2014, the disease has continued to spread within these regions. In 2017, the virus spread further west and the first cases of disease were reported in the Czech Republic and Romania, in wild boar and domestic pigs, respectively. To control further spread, knowledge of different modes of transmission, including indirect transmission via a contaminated environment, is crucial. Up until now, such an indirect mode of transmission has not been demonstrated. In this study, transmission via an environment contaminated with excretions from ASFV ‐infected pigs was investigated. Following euthanasia of pigs that were infected with an isolate of ASFV from Poland (POL /2015/Podlaskie/Lindholm), healthy pigs were introduced into the pens, in which the ASFV ‐infected pigs had been housed. Introduction was performed at 1, 3, 5 or 7 days, following euthanasia of the infected pig groups. Pigs, that were introduced into the contaminated environment after 1 day, developed clinical disease within 1 week, and both ASFV DNA and infectious virus were isolated from their blood. However, pigs introduced into the contaminated pens after 3, 5 or 7 days did not develop any signs of ASFV infection and no viral DNA was detected in blood samples obtained from these pigs within the following 3 weeks. Thus, it was shown that exposure of pigs to an environment contaminated with ASFV can result in infection. However, the time window for transmissibility of ASFV seems very limited, and, within our experimental system, there appears to be a rapid decrease in the infectivity of ASFV in the environment.     

Risk Factors for African Swine Fever in Smallholder Pig Production Systems in Uganda

Smallholder pig production in Uganda is constrained by poor management and high disease burden, with African swine fever (ASF) being one of the most important contributors. However, data to develop appropriate evidence‐based disease mitigating interventions along the pig value chain are lacking. This study aimed at determining risk factors associated with the occurrence of outbreaks of ASF in selected districts. A cross‐sectional survey of 1195 pig‐keeping households in three districts was carried out between April and July 2013. Households were classified into one of three value chain domains (VCDs) based on where the production was located and where most of the products were sold: rural‐rural (R‐R), rural‐urban (R‐U) and urban‐urban (U‐U). Findings revealed that crop farming is the most common primary activity in the R‐R and R‐U VCDs, while pig keeping was the most common primary activity in the U‐U VCDs. Pigs are mostly kept tethered or left to roam in the R‐R and R‐U VCDs, while in the U‐U VCDs, they are mostly confined in corrals. Nearly 20% of the farmers whose farms were hit by an ASF outbreak subsequently sold all their pigs (healthy and sick) to the market in panic. Factors that positively correlated with recent ASF outbreaks were prompt disposal of dead pigs on farms (P < 0.001, OR = 2.3), wild animals present in the village (P < 0.001, OR = 1.7) and farmers sourcing drugs from stockists (P < 0.001, OR = 1.6); while protective factors were the presence of perimeter fences (P = 0.03, OR = 0.5), attendance of farmers at secondary‐school level and above (P < 0.001, OR = 0.6), routine cleaning of the pig pens (P < 0.001, OR = 0.6) and pigs being the only livestock kept by farmer (P = 0.01, OR = 0.7). Given the current situation, there is a need to raise awareness among farmers and other value chain actors of biosecurity measures and create incentives for farmers to report ASF cases.     

Serological Surveillance and Direct Field Searching Reaffirm the Absence of Ornithodoros Erraticus Ticks Role in African Swine Fever Cycle in Sardinia

African swine fever (ASF), one of the most important diseases of swine, has been endemic in the Italian island of Sardinia for more than 35 years. During these decades, several strategies and eradication efforts have been implemented in the island with limited success. Strong climatic and ecological similarities exist between Sardinia and one area of the Iberian Peninsula where Ornithodoros erraticus ticks were involved in the persistence of ASF from 1960 to 1995. This fact leads to the hypothesis that, potentially, Ornithodoros ticks could be also involved in the ASF cycle in Sardinia, thus accounting for some of the reoccurring ASF outbreaks in this island. Initial efforts aimed at detection of Ornithodoros ticks in Sardinia were performed during the 1970s/1980s with no positive results. Accordingly, the absence of Ornithodoros ticks in Sardinia has been generally accepted. However, since a new and reinforced ASF eradication programme has been recently launched in Sardinia, it is essential to clarify the presence and role of these soft ticks in the epizootiology of ASF in this island. For that purpose, 1767 porcine serum samples collected from all around the island (1261 from domestic and 506 from wild boar) were analysed by ELISA for antibodies to salivary antigens of Ornithodoros erraticus. In addition, Ornithodoros ticks were directly searched in a number of pig premises that have suitable habitats for these ticks and were located in areas repeatedly affected by ASF. Only one serum sample resulted positive in the serological assay, and no Ornithodoros ticks were collected in none of the premises. These results indicate that these soft tick species are not involved in the epizootic cycle of ASF in Sardinia and highlight the importance of controlling other risk factors still present in the island for effectively eradicate the disease.     

Thirty‐Five‐Year Presence of African Swine Fever in Sardinia: History,Evolution and Risk Factors for Disease Maintenance

Despite the implementation of control efforts and funds to fight against the disease, African swine fever (ASF) has been present in Sardinia since 1978. It has caused serious problems for both the industrial pig sector and the regional authorities in Sardinia, as well as the economy of Italy and the European Union, which annually supports the costly eradication programme. During this time, ASF has persisted, especially in the central‐east part of Sardinia where almost 75% of the total outbreaks are concentrated. The Sardinian pig sector is clearly divided into two categories based on the specialization and industrialization of production: industrial farms, which represents only 1.8% of the farms in the island and non‐professional holdings, which are comprised of small producers (90% of pig holdings have <15 pigs) and apply little to no biosecurity measures. Additionally, illegally raised pigs are still bred in free‐ranging systems in certain isolated parts of the island, despite strict regulations. The illegal raising of pigs, along with other high‐risk management practices (e.g. use of communal areas) are likely the primary reasons for endemic persistence of the virus in this area. The compensation provided to the farmers, and other aspects of the eradication programme have also negatively influenced eradication efforts, indicating that socio‐cultural and economic factors play an important role in the epidemiology of ASF on the island. The aim of this study was to comprehensively review the evolution of the 35‐year presence of ASF in Sardinia, including control measures, and the environmental and socio‐economic factors that may have contributed to disease endemicity on the island. The present review highlights the need for a coordinated programme that considers these socio‐economic and environmental factors and includes an assessment of new cost‐effective control strategies and diagnostic tools for effectively controlling ASF in Sardinia.     

Diagnostic specificity of the African swine fever virus antibody detection enzyme‐linked immunosorbent assay in feral and domestic pigs in the United States

African swine fever (ASF) is a highly contagious haemorrhagic disease of pigs that has the potential to cause mortality nearing 100% in naïve animals. While an outbreak of ASF in the United States’ pig population (domestic and feral) has never been reported, an introduction of the disease has the potential to cause devastation to the pork industry and food security. During the recovery phase of an outbreak, an antibody detection diagnostic assay would be required to prove freedom of disease within the previously infected zone and eventually nationwide. Animals surviving an ASF infection would be considered carriers and could be identified through the persistence of ASF viral antibodies. These antibodies would demonstrate exposure to the disease and not vaccination, as there is no ASF vaccine available. A well‐established commercial enzyme‐linked immunosorbent assay (ELISA) detects antibodies against ASF virus (ASFV), but the diagnostic specificity of the assay had not been determined using serum samples from the pig population of the United States. This study describes an evaluation of the World Organization for Animal Health (OIE)‐recommended Ingezim PPA COMPAC ELISA using a comprehensive cohort (n = 1791) of samples collected in the United States. The diagnostic specificity of the assay was determined to be 99.4% (95% confidence interval (CI): [98.9, 99.7]). The result of this study fills a gap in understanding the performance of the Ingezim PPA COMPAC ELISA in the ASF naïve pig population of the United States.     

Antigenic regions of African swine fever virus phosphoprotein P30

African swine fever (ASF) is one of the most complex and lethally haemorrhagic viral diseases of swine, affecting all breeds and ages of pigs. In the absence of ASF vaccines, reliable laboratory diagnosis and restricted biosecurity are critical for disease prevention and control. A detection of ASF‐specific antibodies in an unvaccinated pig is a good marker for the diagnosis of ASF. The immunoperoxidase test (IPT) is a sensitive test for detecting ASF virus (ASFV) antibodies. However, due to the complexity of the procedure, the IPT is only suitable to be used as a confirmatory test. The ASFV p30 protein‐based enzyme‐linked immunosorbent assay (ELISA) is widely used for ASFV antibody screening, but the sensitivity is not comparable to the IPT. It is essential to have a better understanding of the antigenic properties of ASFV p30 to improve p30‐based serologic tests. In this study, we developed a panel of 21 monoclonal antibodies (mAbs) against ASFV p30. With 14 out of the 21 mAbs, we defined 4 antigenic regions that contain at least 4 linear epitopes. Nine of the 14 mAbs mapped to antigenic regions 3 and 4 reacted with p30 in all serologic methods tested in this study, such as indirect immunofluorescence assay (IFA), ELISA and Western blot. The antigenic regions 3 and 4 are highly conserved and immunodominant in host antibody response. These mAbs and the defined p30 antigenic regions 3 and 4 provide valuable tools for the development and improvement of ASF serologic assays.     

Infection of pigs with African swine fever virus via ingestion of stable flies (Stomoxys calcitrans)

Within Eastern Europe, African swine fever virus (ASFV ) has unexpectedly spread to farms with high biosecurity. In an attempt to explain this process, pigs were allowed to ingest flies that had fed on ASFV ‐spiked blood, which had a realistic titre for an infected pig. Some of the pigs became infected with the virus. Thus, ingestion of blood‐sucking flies, having fed on ASFV ‐infected wild boar before entering stables, represents a potential route for disease transmission.     

Detection of African Swine Fever Antibodies in Experimental and Field Samples from the Russian Federation: Implications for Control

African swine fever (ASF) re‐entered in Europe in 2007 by Georgia rapidly affecting neighbouring countries. Since then, ASF has caused severe problems to the Russian Federation (RF) and spread to Northern and Western regions, including Ukraine (2012 and 2014) and Belarus (2013). At the beginning of 2014, dead wild boars were found in Lithuania and Poland. Several outbreaks have been later notified in the European Union(EU), affecting domestic pigs and wild boar of Latvia, Lithuania and Poland, and also wild boar in Estonia, causing major problems for the EU pig sector. Some studies have been performed with this ASFV isolate, revealing that it belongs to genotype II and causes an acute form of the disease. However, few data are available about the presence of antibodies in field and experimental samples from the affected area. This study analysed samples from experimental infections with ASFV isolated from the RF in 2013 (74 sera and 3 tissue exudates), and field samples from the RF from 2013 to 2014 (266 samples, including 32 and 7 tissue exudates from domestic pigs and wild boar, respectively). All samples were tested by a commercial ELISA and, some of them (79), also by immunochromatographic tests. Positive and doubtful samples were confirmed by immunoblotting test. Positive results were found in experimental and field samples, which confirm the presence of antibodies against ASFV in the RF. Antibodies were detected in animals inoculated with three different ASFV isolates, with some differences found among them. Only a small percentage of field samples was positive for ASF antibodies (3.7%), in agreement with other observations that reported a high virulence for the ASFV isolates in the area. These results confirm the potential presence of survivor animals that should be considered in affected areas to help design effective control and eradication plans against ASF.     

Recombinant Protein p30 for Serological Diagnosis of African Swine Fever by Immunoblotting Assay

This article is devoted to the development and evaluation of the immunoblotting test system for serological diagnosis of African swine fever (ASF), based on the highly purified recombinant p30 of ASF virus (ASFV) strain Stavropol 01/08 (Stavropol 2008), representative of the ASFV currently circulating in the Russian Federation. The main project stages are as follows: (i) cloning of the central hydrophilic region of the ASFV gene CP204L (p30) into a prokaryotic vector; (ii) expression and chromatographic purification of the recombinant product p30 with thioredoxin and poly‐histidine site (p30e1_TrxA_6xHis); (iii) development of the immunoblotting test system (Rec p30‐IB) using the highly purified recombinant p30; and (iv) evaluation of Rec p30‐IB using sera and organ samples from domestic pigs and wild boars experimentally or naturally infected by ASFV. Testing of the Rec p30‐IB showed the diagnostic specificity and sensitivity of the assay to be 98.75% and 100.00%, respectively. High sensitivity of the Rec p30‐IB allowed the detection of ASFV‐specific antibodies in samples of organs of the immune system and blood sera, collected from domestic pigs and wild boars, starting from 6 to 8 days post‐infection, regardless of virus virulence, seroimmunotype and geographic origin of the samples (East Europe, South Europe, West Europe, Central and south‐east Africa).     

Seroprevalence of Rift valley fever in South African domestic and wild suids (1999–2016)

Rift valley fever (RVF) is a vector‐borne viral disease of domestic ruminants, camels and man, characterized by widespread abortions and neonatal deaths in animals, and flu‐like symptoms, which can progress to hepatitis and encephalitis in humans. The disease is endemic in Africa, Saudi Arabia and Yemen, and outbreaks occur after periods of high rainfall, or in environments supporting the proliferation of RVF virus (RVFV)‐infected mosquito vectors. The domestic and wild animal maintenance hosts of RVFV, which may serve as sources of virus during inter‐epidemic periods (IEPs) and contribute to occurrence of sporadic outbreaks, remain unknown, although reports indicate that the African buffalo (Syncerus caffer) may play a role. Due to the close proximity of the habitats of domestic pigs and warthogs to those of known domestic and wild ruminant RVFV maintenance hosts respectively, our study investigated their possible role in the epidemiology of RVF in South Africa by evaluating RVFV exposure and seroconversion in suids. A total of 107 warthog and 3,984 domestic pig sera from 2 and all 9 provinces of South Africa, respectively, were screened for presence of RVFV neutralizing antibodies using the virus neutralization test (VNT). Sero‐positivity rates of 1.87% (95% CI: 0.01%–6.9%) and 0.68% (95% CI: 0.49%–1.04%) were observed for warthogs and domestic pigs, respectively, but true prevalence rates, taking test sensitivity and specificity into account, were lower for both groups. There was a strong association between the results of the two groups (χ² = 0.75, p = .38), and differences in prevalence between the epidemic and IEPs were non‐significant for all suid samples tested (p > .05). This study, which provides the first evidence of probable exposure and infection of South African domestic pigs and warthogs to RVFV, indicates that further investigations are warranted, to fully clarify the role of suids in the epidemiology of RVF.     

First Detection of Antibodies Against African Swine Fever Virus in Faeces Samples

African swine fever (ASF) is a viral, highly lethal haemorrhagic disease of swine with no available vaccine or effective treatment. Introduction of ASF into a country triggers immediate restriction measures that cause significant economic losses and threatens spread to neighbouring countries. Wild boar populations have been recently assigned an essential role in the spread of African swine fever virus (ASFV) to European countries. Therefore, effective surveillance and monitoring of wild boar populations is required, but sampling wild boar is logistically challenging and expensive. This study assessed the feasibility of detecting antibodies against ASFV in faeces for later implementation in surveillance and control programmes. Two groups of pigs were experimentally infected with an attenuated ASFV isolate Ken05, and blood, oral fluid and faecal samples were tested for the presence of viral DNA using quantitative real‐time polymerase chain reaction (qPCR) to monitor infection progress. Faecal samples were analysed using two indirect enzyme‐linked immunosorbent assays (ELISAs) based on semipurified viral protein (vp) 72 or purified recombinant vp30 expressed in mammalian cells. Faecal samples from 9 of 10 pigs with non‐haemorrhagic diarrhoea tested positive for antibodies against ASFV using the two ELISA tests that showed a positive correlation. The serum sample results from the two indirect ELISAs were compared against results from the reference ELISA technique and the immunoperoxidase test. Our findings indicate the feasibility of faecal sampling for detecting anti‐ASFV antibodies, which may provide a practical non‐invasive alternative for sampling wild boar populations. In conclusion, the application of these ELISA tests to faecal field samples could be particularly useful to screen for the presence of ASF in field conditions.