An Updated Review of Ornithodoros Ticks as Reservoirs of African Swine Fever in Sub-Saharan Africa and Madagascar.

This updated review provides an overview of the available information on Ornithodoros ticks as reservoirs and biological vectors of the ASF virus in Africa and Indian Ocean islands in order to update the current knowledge in this field, inclusive of an overview of available methods to investigate the presence of ticks in the natural environment and in domestic pig premises. In addition, it highlights the major areas of research that require attention in order to guide future investigations and fill knowledge gaps. The available information suggests that current knowledge is clearly insufficient to develop risk-based control and prevention strategies, which should be based on a sound understanding of genotype distribution and the potential for spillover from the source population. Studies on tick biology in the natural and domestic cycle, including genetics and systematics, represent another important knowledge gap. Considering the rapidly changing dynamics affecting the African continent (demographic growth, agricultural expansion, habitat transformation), anthropogenic factors influencing tick population distribution and ASF virus (ASFV) evolution in Africa are anticipated and have been recorded in southern Africa. This dynamic context, together with the current global trends of ASFV dissemination, highlights the need to prioritize further investigation on the acarological aspects linked with ASF ecology and evolution.

Innovative Research Offers New Hope for Managing African Swine Fever Better in Resource-Limited Smallholder Farming Settings: A Timely Update.

African swine fever (ASF) in domestic pigs has, since its discovery in Africa more than a century ago, been associated with subsistence pig keeping with low levels of biosecurity. Likewise, smallholder and backyard pig farming in resource-limited settings have been notably affected during the ongoing epidemic in Eastern Europe, Asia, the Pacific, and Caribbean regions. Many challenges to managing ASF in such settings have been identified in the ongoing as well as previous epidemics. Consistent implementation of biosecurity at all nodes in the value chain remains most important for controlling and preventing ASF. Recent research from Asia, Africa, and Europe has provided science-based information that can be of value in overcoming some of the hurdles faced for implementing biosecurity in resource-limited contexts. In this narrative review we examine a selection of these studies elucidating innovative solutions such as shorter boiling times for inactivating ASF virus in swill, participatory planning of interventions for risk mitigation for ASF, better understanding of smallholder pig-keeper perceptions and constraints, modified culling, and safe alternatives for disposal of carcasses of pigs that have died of ASF. The aim of the review is to increase acceptance and implementation of science-based approaches that increase the feasibility of managing, and the possibility to prevent, ASF in resource-limited settings. This could contribute to protecting hundreds of thousands of livelihoods that depend upon pigs and enable small-scale pig production to reach its full potential for poverty alleviation and food security.

Retrospective Multi-Locus Sequence Analysis of African Swine Fever Viruses by "PACT" Confirms Co-Circulation of Multiple Outbreak Strains in Uganda.

African swine fever (ASF) is a haemorrhagic fever of swine that severely constrains pig production, globally. In Uganda, at least 388 outbreaks of ASF were documented from 2001 to 2012. We undertook a retrospective serological and molecular survey of ASF virus (ASFV) using banked samples collected from seven districts (Pallisa, Lira, Abim, Nebbi, Kabarole, Kibaale, and Mukono) of Uganda. Six assays (ELISA for antibody detection, diagnostic p72 gene PCR and genomic amplification, and sequencing of four gene regions (p72 [P], p54 [A], CVR of the 9RL-ORF [C], and TK [T]), hereinafter referred to as P-A-C-T (PACT)) were evaluated. Antibodies to ASFV were detected in the Abim district (6/25; 24.0%), and the remainder of the serum samples were negative (187/193; 96.9%). For the tissue samples, ASFV detection by assay was 8.47% for P, 6.78% for A, 8.47% for C, and 16.95% for T. The diagnostic PCR (p72 gene) detected seven positive animals from four districts, whereas the TK assay detected ten positives from all seven districts. In addition to the superior detection capability of TK, two virus variants were discernible, whereas CVR recovered three variants, and p72 and p54 sequencing each identified a single variant belonging to genotype IX. Our results indicate that dependence on serology alone underestimates ASF positivity in any infected region, that multi-locus sequence analysis provides better estimates of outbreak strain diversity, and that the TK assay is superior to the WOAH-prescribed conventional p72 diagnostic PCR, and warrants further investigation.

Review of the Pig-Adapted African Swine Fever Viruses in and Outside Africa.

The region in eastern, central and southern Africa (ECSA) where African swine fever (ASF) originated in a sylvatic cycle is home to all the p72 genotypes of ASF virus identified so far. While 20 of the 24 genotypes have been isolated from outbreaks in domestic pigs in the region, only five of the genotypes (I, II, VIII, IX, X) have an extended field presence associated with domestic pigs. Of the genotypes that appear to be strongly adapted to domestic pigs, two have spread beyond the African continent and have been the focus of efforts to develop vaccines against ASF. Most of the experimental ASF vaccines described do not protect against a wider spectrum of viruses and may be less useful in the event of incursions of different strains or where multiple genotypes co-exist. The other three pig-adapted strains that are currently restricted to the ECSA region might spread, and priority should be given to understanding not only the genetic and antigenic characteristics of these viruses but also their history. We review historic and current knowledge of the distribution of these five virus genotypes, and note that as was the case for genotype II, some pig-associated viruses have the propensity for geographical range expansion. These features are valuable for prioritizing vaccine-development efforts to ensure a swift response to virus escape. However, whilst ASF vaccines are critical for high-production systems, global food security relies on parallel efforts to improve biosecurity and pig production in Africa and on continued ASFV surveillance and characterisation in the ECSA region.

Live Attenuated African Swine Fever Viruses as Ideal Tools to Dissect the Mechanisms Involved in Cross-Protection.

African swine fever (ASF) has become the major threat for the global swine industry. Furthermore, the epidemiological situation of African swine fever virus (ASFV) in some endemic regions of Sub-Saharan Africa is worse than ever, with multiple virus strains and genotypes currently circulating in a given area. Despite the recent advances on ASF vaccine development, there are no commercial vaccines yet, and most of the promising vaccine prototypes available today have been specifically designed to fight the genotype II strains currently circulating in Europe, Asia, and Oceania. Previous results from our laboratory have demonstrated the ability of BA71∆CD2, a recombinant LAV lacking CD2v, to confer protection against homologous (BA71) and heterologous genotype I (E75) and genotype II (Georgia2007/01) ASFV strains, both belonging to same clade (clade C). Here, we extend these results using BA71∆CD2 as a tool trying to understand ASFV cross-protection, using phylogenetically distant ASFV strains. We first observed that five out of six (83.3%) of the pigs immunized once with 106 PFU of BA71∆CD2 survived the tick-bite challenge using Ornithodoros sp. soft ticks naturally infected with RSA/11/2017 strain (genotype XIX, clade D). Second, only two out of six (33.3%) survived the challenge with Ken06.Bus (genotype IX, clade A), which is phylogenetically more distant to BA71∆CD2 than the RSA/11/2017 strain. On the other hand, homologous prime-boosting with BA71∆CD2 only improved the survival rate to 50% after Ken06.Bus challenge, all suffering mild ASF-compatible clinical signs, while 100% of the pigs immunized with BA71∆CD2 and boosted with the parental BA71 virulent strain survived the lethal challenge with Ken06.Bus, without almost no clinical signs of the disease. Our results confirm that cross-protection is a multifactorial phenomenon that not only depends on sequence similarity. We believe that understanding this complex phenomenon will be useful for designing future vaccines for ASF-endemic areas.

Investigation into eradication of African swine fever in domestic pigs from a previous outbreak (2016/17) area of South Africa.

A serological survey was conducted to evaluate the eradication of African swine fever (ASF) infection eighteen months after clinical surveillance and selective culling had been completed during domestic cycle outbreaks in parts of South Africa in 2016/17. Three hundred and twenty-two serum samples from 85 pig keepers were collected in the study area and tested for the presence of antibodies against the ASF virus (ASFV). None of the samples contained detectable levels of antibodies against ASFV. These results together with the findings from clinical surveillance following culling activities suggest that the disease had been eradicated from the domestic pig population in this area following the outbreaks. Questionnaire responses from the pig keepers in this area highlighted the need to implement basic biosecurity measures in smallholder pig keepers to prevent outbreaks of ASF in South Africa.

Investigation of African swine fever outbreaks in pigs outside the controlled areas of South Africa, 2012-2017.

South Africa historically experienced sporadic African swine fever (ASF) outbreaks in domestic pigs in the northern parts of the country. This was subsequently indicated to be because of spillover from the sylvatic cycle of ASF between warthog and tampans (soft ticks) in the area. South Africa declared this area an ASF-controlled area in 1935, and the area is still controlled in terms of the Animal Diseases Act, 1984 (Act 35 of 1984). Two main epidemics of ASF in domestic pigs were identified outside of the South African ASF-controlled area. The first occurred in 2012 with outbreaks in Gauteng and Mpumalanga provinces, and the second occurred in 2016-2017 with outbreaks in the North West, Free State and Northern Cape provinces. These were the first ASF epidemics in South Africa associated with transmission of the disease via a domestic cycle. This study found that the spread of ASF in these epidemics was mainly via auctions, swill feeding and scavenging. These three aspects need to be addressed in terms of awareness and education on the disease including implementation of biosecurity measures in order to prevent future ASF outbreaks in South Africa. Specific biosecurity measures should be implemented in the semi-commercial sector to prevent ASF-infected pigs and pig products from being moved to naïve pigs and therefore spreading the disease.

Understanding African swine fever outbreaks in domestic pigs in a sylvatic endemic area: The case of the South African controlled area between 1977-2017.

South Africa declared a controlled area for African swine fever (ASF) in 1935, consisting of the northern parts of Limpopo, Mpumalanga, North West and Kwa-Zulu Natal Provinces. The area was delineated based on the endemic presence of the sylvatic cycle of ASF, involving warthogs and argasid ticks. Occasionally, spillover occurs from the sylvatic cycle to domestic pigs, causing ASF outbreaks. In the period 1977 to 2017, 59 outbreaks of ASF were reported in domestic pigs within the ASF controlled area of South Africa. During these outbreaks, at least 4,031 domestic pigs either died or were culled. Season did not affect the number of reported ASF outbreaks, but the number of reported outbreaks in this area per year was thought to be slowly increasing, although not statistically significant. Outbreaks occurred predominantly in Limpopo province (93%) and were mostly due to contact (or suspected contact) with warthog or warthog carcasses. Clustering analysis of outbreaks found that the local municipalities of Ramotshere Moiloa, Lephalale and Thabazimbi had the highest relative risk for outbreaks. In 32 of the 59 outbreaks, the genotype of the ASF virus (ASFV) involved could be determined. Phylogenetic analysis of ASFVs detected in domestic pigs during the study period revealed that p72 genotypes I, III, IV, VII, VIII, XIX, XX, XXI and XXII had been involved in causing outbreaks within the ASF controlled area. No outbreaks were reported in the Kwa-Zulu Natal part of the controlled area during this period. South Africa is unlikely to eradicate all sources of ASFV as spillover from the sylvatic cycle in the controlled area continued to occur, but with the implementation of appropriate biosecurity measures pigs can be successfully farmed despite the presence of ASFV in African wild suids and soft ticks.

Characterization of SAT2 foot-and-mouth disease 2013/2014 outbreak viruses at the wildlife-livestock interface in South Africa.

The Southern African Territories (SAT)-type foot-and-mouth disease viruses (FMDV) are endemic to the greater Kruger National Park (KNP) area in South Africa, where they are maintained through persistent infections in African buffalo. The occurrence of FMDV within the Greater KNP area constitutes a continual threat to the livestock industry. To expand on knowledge of FMDV diversity, the genetic and antigenic relatedness of SAT2-type viruses isolated from cattle during a FMD outbreak in Mpumalanga Province in 2013 and 2014 were investigated. Cattle from twelve diptanks tested positive on polymerase chain reaction (PCR), and molecular epidemiological relationships of the viruses were determined by VP1 sequencing. Phylogenetic analysis of the SAT2 viruses from the FMD outbreak in Mpumalanga in 2013/2014 revealed their genetic relatedness to other SAT2 isolates from topotype I (South Africa, Zimbabwe and Mozambique), albeit genetically distinct from previous South African outbreak viruses (2011 and 2012) from the same topotype. The fifteen SAT2 field isolates clustered into a novel genotype with ≥98.7% nucleotide identity. High neutralization antibody titres were observed for four 2013/2014 outbreak viruses tested against the SAT2 reference antisera representative of viruses isolated from cattle and buffalo from South Africa (topotype I) and Zimbabwe (topotype II). Comparison of the antigenic relationship (r1 values) of the outbreak viruses with reference antisera indicated a good vaccine match with 90% of r1 values > 0.3. The r1 values for the 2013/2014 outbreak viruses were 0.4 and above for the three South African vaccine/reference strains. These results confirm the presence of genetic and antigenic variability in SAT2 viruses and suggest the emergence of new variants at the wildlife-livestock interface in South Africa. Continuous characterization of field viruses should be performed to identify new virus strains as epidemiological surveillance to improve vaccination efforts.

The Epidemiology of African Swine Fever in "Nonendemic" Regions of Zambia (1989-2015): Implications for Disease Prevention and Control.

African swine fever (ASF) is a highly contagious and deadly viral hemorrhagic disease of swine. In Zambia, ASF was first reported in 1912 in Eastern Province and is currently believed to be endemic in that province only. Strict quarantine measures implemented at the Luangwa River Bridge, the only surface outlet from Eastern Province, appeared to be successful in restricting the disease. However, in 1989, an outbreak occurred for the first time outside the endemic province. Sporadic outbreaks have since occurred almost throughout the country. These events have brought into acute focus our limited understanding of the epidemiology of ASF in Zambia. Here, we review the epidemiology of the disease in areas considered nonendemic from 1989 to 2015. Comprehensive sequence analysis conducted on genetic data of ASF viruses (ASFVs) detected in domestic pigs revealed that p72 genotypes I, II, VIII and XIV have been involved in causing ASF outbreaks in swine during the study period. With the exception of the 1989 outbreak, we found no concrete evidence of dissemination of ASFVs from Eastern Province to other parts of the country. Our analyses revealed a complex epidemiology of the disease with a possibility of sylvatic cycle involvement. Trade and/or movement of pigs and their products, both within and across international borders, appear to have been the major factor in ASFV dissemination. Since ASFVs with the potential to cause countrywide and possibly regional outbreaks, could emerge from "nonendemic regions", the current ASF control policy in Zambia requires a dramatic shift to ensure a more sustainable pig industry.

Reemergence of African Swine Fever in Zimbabwe, 2015.

Zimbabwe is the only country in southern Africa with no reported African swine fever (ASF) outbreaks during 1993-2014. However, the 2015 discovery of genotype II ASF virus in Zimbabwe indicates the reemergence of ASF in this country and suggests that this viral genotype may be spreading through eastern and southern Africa.

Incidence of adenoviruses in raw and treated water.

Adenoviruses are of major public health importance and are associated with a variety of clinical manifestations, i.e. gastroenteritis, eye infections and respiratory infections. The importance of water in the epidemiology of adenoviruses and the potential health risks constituted by adenoviruses in water sources and supplies are widely recognised. This study was conducted to assess the incidence of human adenoviruses in raw and treated water systems. Various raw and treated water were routinely monitored for the presence of adenoviruses, over a 1-year period (July 2000-June 2001). The supplies were derived from acceptable quality surface water sources using treatment processes, which conform to international standards for the production of safe drinking water. Adenoviruses were detected by firstly amplifying the viruses in cell cultures and then amplifying the extracted nucleic acids of these viruses using molecular techniques (nested PCR). The results indicated human adenoviruses present in 13 (12.75%) of the raw and 9 (4.41%) of the treated water samples tested. The combination of cell culture and nested PCR has proved to be a quick and reliable method for the detection of adenoviruses in water environments.