Genetic characterisation of African swine fever viruses from outbreaks in southern Africa (1973-1999)

African swine fever (ASF) is a highly lethal and economically significant disease of domestic pigs in the southern African sub-region, where outbreaks regularly occur. There is anecdotal evidence suggesting that trans-boundary movement of infected animals may have played a role in precipitating widespread outbreaks in the past, however, since the 1970s outbreaks have generally been more localised, particularly in those countries where control of animal movement is strictly regulated. The origin and relatedness of regional ASF outbreaks was investigated here by means of a two-step genetic characterisation approach whereby p72 gene sequencing was used to delineate genotypes, prior to intra-genotypic resolution of viral relationships by central variable region (CVR) characterisation of the 9RL ORF. In this manner, regional virus heterogeneity and epidemiological links between outbreaks could be assessed for the first time through phylogenetic analysis of the C-terminal end of the p72 gene of viruses recovered from domestic pig outbreaks in southern Africa between 1973 and 1999. The phylogeny revealed the presence of 14 distinct p72 genotypes of which 6 (genotypes XVII-XXII) were considered novel. Eight of these were country-specific with the remaining six having a trans-boundary distribution. CVR products were heterogeneous in size ranging from 377bp to 533bp across the 14 southern African genotypes. Within-genotype CVR comparisons revealed the presence of a genotype XIX virus with an extended field presence in South Africa (1985-1996) and permitted discrimination between three genotype VII viruses that were identical across the p72 gene.     

Molecular characterization of African swine fever virus from domestic pigs in northern Tanzania during an outbreak in 2013

African swine fever (ASF) is an acute, highly contagious and deadly viral hemorrhagic fever of domestic pigs caused by African swine fever virus (ASFV), a double-stranded DNA virus of the family Asfarviridae. In this study, molecular diagnosis and characterization of outbreak ASFV in northern Tanzania, was performed on spleen, lymph node, kidney, and heart samples collected in June and July 2013 from domestic pigs that died during a hemorrhagic disease outbreak. Confirmatory diagnosis of ASF was performed using polymerase chain reaction (PCR) by partial amplification of B646L gene of ASFV encoding the major capsid protein p72 using PPA1/PPA2 primers. PCR using PPA1/PPA2 primers produced an expected PCR product size, confirming ASF outbreak in northern Tanzania. In addition, nucleotide amplification and sequencing, and phylogenetic reconstruction of the variable 3′-end of the B646L gene and complete E183L gene encoding the inner envelope transmembrane protein p54 showed that the 2013 outbreak ASFV from northern Tanzania were 100 % identical and clustered into ASFV B646L (p72) and E183L (p54) genotype X. Furthermore, the tetrameric amino acid repeats within the central variable region (CVR) of the B602L gene coding for the J9L protein had the signature BNBA(BN)₅NA with a single novel tetramer NVDI (repeat code N). The results of the present study confirm an ASF outbreak in northern Tanzania in the year 2013 and show that the present outbreak ASFV is closely related to other ASFV from ticks, warthogs, and domestic pigs previously reported from Tanzania.     

Review of African swine fever outbreaks history in South Africa: From 1926 to 2018

The article reviews the outbreaks and distribution of African swine fever (ASF) in South Africa since the first probable outbreak that occurred in the Koedoesrand Ward in 1926. Retrospective data on the ASF outbreaks in South Africa were obtained from the World Organisation for Animal Health (OIE) disease database and the South African veterinary services annual reports in addition to published articles and online sources. South Africa has experienced many outbreaks that can be divided into 2 time periods: the period before the development of the OIE diseases database (1993) and the period after. More than 141 outbreaks of ASF were reported during the first period. Since the development of OIE disease database, 72 outbreaks directly involving 2968 cases, 2187 dead and 2358 killed pigs mainly in smallholder pig farms were reported. The median number of cases for a given ASF outbreak is 17, but in 50% of outbreaks no pigs were killed for prevention. The most important ASF outbreak was reported in April 2014 in the Greater Zeerust district (North West province) involving 326 cases and 1462 killed pigs. However, the outbreak with highest mortality involving 250 pigs was reported in 2016 (Free State province). According to phylogenetic analysis, nine p72 genotypes (I, III, IV, VII, VIII, XIX, XX, XXI and XXII) have been identified in South Africa. Season-wise, more outbreaks were recorded during summer. It was also observed that the OIE disease database could contain errors that would have been introduced through compiled forms at country level. Spatiotemporal studies on ASF outbreaks in South Africa are therefore required in order to assess statistically and quantitatively the clustering of outbreaks over space and time.     

非洲猪瘟对我国养猪业的影响与防控建议

非洲猪瘟是由非洲猪瘟病毒引起的一种高度接触传染性、广泛出血性猪病毒病,死亡率可高达100%,无商品化疫苗可用于其预防。自8月初我国爆发首例非洲猪瘟疫情以来,截至11月5日,短短三个月时间里,已有14个省累计爆发58起疫情,扑杀生猪近50万头,直接经济损失数十亿元,给我国养猪业造成了巨大冲击。面对前所未有的非洲猪瘟疫情,我国政府的应对是果断有力的,但也面临种种挑战。本文对我国非洲猪瘟疫情形势及其对相关行业的影响进行了分析,总结了非洲猪瘟防控的重点和难点,并针对目前的形势,提出了遏制非洲猪瘟疫情进一步蔓延的建议和对策。     

African swine fever in Mozambique: review, risk factors and considerations for control

African swine fever (ASF) is the most important disease that constrains pig production in Mozambique. Until 1994 it was apparently restricted to the central and northern provinces, but since 1994 outbreaks have been experienced throughout the country. ASF causes severe economic losses both in the small commercial sector and among the large numbers of small-scale producers in the family sector in rural and peri-urban areas. The history of ASF in Mozambique since its first confirmation in 1960 is briefly reviewed, recent outbreaks are reported, and the available information on the virus genotypes that have been responsible for some of the outbreaks is presented. Epidemiological factors that contribute to ASF outbreaks and strategies for limiting the negative effects of the disease in the different pig farming sectors in Mozambique, including raising farmer and community awareness, are discussed.     

Pathological lesions and presence of viral antigens in four surviving pigs in African swine fever outbreak farms in Vietnam

Investigation of the role of animals that have recovered and survived from African swine fever (ASF) in carrying the ASF virus is currently intense and ongoing. However, no clear definition of the carrier stage has been established. The aim of the present study was to establish criteria to elucidate a clear status of survival in naturally ASF-infected domestic pigs in Vietnam. Seroconversion from previous infection was confirmed by serological assay, and the absence of the viral genome in various organs was also assured by molecular analysis of a partial p72 gene. We recognized that histopathological evidence could benefit from further insights into the status and role of the surviving animals; therefore, we performed a histopathological study on four pigs from farms with a history of ASF outbreak. We found fibrotic changes in the reparative process as the main finding in all four pigs. Immunohistochemical detection of viral protein revealed an interesting result. Despite the negative result from viral genome detection, the p30 protein gave a positive signal in the tonsils, lung, and stomach. This raises the possibility of stress-induced viral reactivation in long-term survivors and the risk of further outbreaks from human handling of contaminated carcasses.     

African swine fever outbreak on a medium-sized farm in Uganda: biosecurity breaches and within-farm virus contamination

In Uganda, a low-income country in east Africa, African swine fever (ASF) is endemic with yearly outbreaks. In the prevailing smallholder subsistence farming systems, farm biosecurity is largely non-existent. Outbreaks of ASF, particularly in smallholder farms, often go unreported, creating significant epidemiological knowledge gaps. The continuous circulation of ASF in smallholder settings also creates biosecurity challenges for larger farms. In this study, an on-going outbreak of ASF in an endemic area was investigated on farm level, including analyses of on-farm environmental virus contamination. The study was carried out on a medium-sized pig farm with 35 adult pigs and 103 piglets or growers at the onset of the outbreak. Within 3 months, all pigs had died or were slaughtered. The study included interviews with farm representatives as well as biological and environmental sampling. ASF was confirmed by the presence of ASF virus (ASFV) genomic material in biological (blood, serum) and environmental (soil, water, feed, manure) samples by real-time PCR. The ASFV-positive biological samples confirmed the clinical assessment and were consistent with known virus characteristics. Most environmental samples were found to be positive. Assessment of farm biosecurity, interviews, and the results from the biological and environmental samples revealed that breaches and non-compliance with biosecurity protocols most likely led to the introduction and within-farm spread of the virus. The information derived from this study provides valuable insight regarding the implementation of biosecurity measures, particularly in endemic areas.     

Newcastle disease virus isolated from recent outbreaks in Taiwan phylogenetically related to viruses (genotype VII) from recent outbreaks in western Europe

Three major outbreaks of Newcastle disease (ND) occurred in Taiwan in the last three decades (in 1969, 1984, and 1995). Newcastle disease viruses (NDVs) isolated in the three outbreaks, together with those isolated in 1998, were sequenced between nucleotides 47 and 435 of the fusion gene. A phylogenetic tree based on sequences obtained showed that the NDV isolated in 1969 was similar to the genotype III viruses. In contrast, all isolates in 1984 and seven of the eight isolates in 1995, together with all isolates in 1998, fell into the genotype VII. These results suggest that the 1969 outbreak of ND in Taiwan was caused by the genotype III virus, whereas the 1984 and 1995 outbreaks were caused by the genotype VII viruses. To date, the genotype VII viruses have caused many outbreaks in east Asia and western Europe. We suspect that these outbreaks have constituted the fourth panzootic of ND, which is distinct from the third panzootic caused by the "pigeon PMV-1 viruses." NDV isolated in Taiwan in 1984 was the earliest isolation of the genotype VII virus.     

Antigenic and genetic characterisation of Newcastle disease viruses isolated from outbreaks in domestic fowl and turkeys in Great Britain during 1997

Antigenic and genetic analyses of viruses from the 11 outbreaks of Newcastle disease in Great Britain, 12 of the outbreaks in Northern Ireland and the single outbreak in the Republic of Ireland which occurred in 1997, indicated that they were all essentially similar. In addition, the viruses from the British Isles were very similar to viruses isolated from three outbreaks in pheasants in Denmark between August and November 1996, from a goosander in Finland in September 1996, from an outbreak in chickens in Norway in February 1997, and from an outbreak in chickens in Sweden in November 1997. Viruses from outbreaks in other countries during 1995 to 1997 could be distinguished antigenically and/or genetically from the 1996 to 1997 Scandinavian/British Isles isolates, as could viruses responsible for two separate outbreaks in caged birds in quarantine premises in Great Britain in March 1997. Minor nucleotide differences in the 413-base region of the fusion gene and the 187-base region of the haemagglutinin-neuraminidase gene sequenced in this study allowed the 1996 to 1997 Scandinavian/British Isles isolates to be divided into groups. These groups broadly corresponded to the clusters of disease outbreaks, but suggested that the discrete outbreak in Scotland was probably the result of virus spread from Northern Ireland. Overall, the antigenic and genetic analyses of these viruses were consistent with the theory that the virus was introduced into the British Isles by migratory birds moving from north-east Europe. However, it was not possible to rule out other sources, such as the movement of pheasants from Denmark.     

Comparison of Pseudomonas aeruginosa isolates from mink by serotyping and pulsed-field gel electrophoresis

Isolates of Pseudomonas aeruginosa from clinical infections in mink were subjected to serotyping and pulsed-field gel electrophoresis (PFGE) using SpeI. A total of 212 isolates of P. aeruginosa from the year 1998 to 2001 were included in this study: 168 isolates from mink obtained from 74 farm outbreaks of haemorrhagic pneumonia. Isolates from mink were separated into 34 distinct clones by PFGE subtyping. All isolates from mink infected during the same farm outbreak were identical, except in one case where two different strains were isolated from mink obtained from the same farm outbreak. P. aeruginosa of specific PFGE types were found to cause clusters of outbreaks on several farms within a few weeks of each other. However, PFGE types of strains causing clusters of farm outbreaks changed from year to year. These results suggest that some outbreaks of haemorrhagic pneumonia are caused by pathogenic strains of P. aeruginosa spread between farms and animals either mechanically, or through feed or water from a common source, rather than by random nosocomial infections with strains from the farm environment.     

Serum concentrations of C-reactive protein, serum amyloid A, and haptoglobin in pigs inoculated with African swine fever or classical swine fever viruses

OBJECTIVE: To determine serum concentrations of the selected acute-phase proteins (APPs) haptoglobin, serum amyloid A (SAA), and C-reactive protein (CRP) in pigs experimentally inoculated with classical swine fever (CSF) and African swine fever (ASF) viruses. ANIMALS: 8 crossbred (Large White x Landrace) 10-week-old pigs. PROCEDURES: Pigs were allocated to 2 groups (4 pigs/group). One group was inoculated with the CSF virus Alfort 187 strain, whereas the other groupwas inoculated with the ASF virus Spain 70 isolate. Blood samples were collected at various time points. At the end of the study, pigs were euthanized and a complete necropsy was performed, including histologic and immunohistochemical analyses. RESULTS: Serum concentrations of APPs increased in pigs inoculated with CSF and ASF viruses, which suggested an acute-phase response in the course of both diseases. The most noticeable increase in concentration was recorded for SAA in both groups (up to a 300-fold increase for CSF virus and an approx 40-fold increase for ASF virus), followed by CRP and then haptoglobin, which each had only 3- to 4-fold increases. CONCLUSIONS AND CLINICAL RELEVANCE: Serum concentrations of APPs increased significantly in pigs inoculated with CSF and ASF viruses. However, differences were evident in serum concentrations of the proteins evaluated in this study.     

African swine fever among slaughter pigs in Mubende district, Uganda

Owing to frequent reports of suspected outbreaks and the presence of reservoir hosts and vectors (warthogs, bushpigs and O. moubata ticks), African swine fever (ASF) is believed to be an endemic disease in Uganda. There have, however, been very few studies carried out to confirm its existence in Uganda. This study was carried out to describe the prevalence of ASF based on pathologic lesions and analysis of serum samples from slaughtered pigs during a suspected outbreak in the Mubende district of Uganda. The study was based on visits to 22 slaughterhouses where individual pigs were randomly selected for a detailed ante-mortem and post-mortem inspections. Sera were also collected for laboratory analysis. A total of 997 pigs (53.7% male and 46.3% female) were examined for lesions suggestive of ASF and sero-positivity of sera for ASF antibodies. The sera were tested using enzyme-linked immunosorbent assay (ELISA) and positive samples were further confirmed with an immunoblot assay. The results showed that 3.8% (38/997) of the pigs examined had clinical signs and post-mortem lesions suggestive of ASF. Two of 997 (0.2%) sera analysed were positive for ASF antibodies. Of the sub-counties investigated, Bagezza (12%) and Kiyuni (11%) had the highest prevalence of lesions suggestive of ASF based on ante- and post-mortem examination results, while Mubende town council (1.7%) had the lowest. This study found a low number of pigs (3.8%) with lesions suggestive of ASF at slaughter and an even lower number of pigs (0.2%) that were seropositive at slaughter, however a significantly higher number of pigs were slaughtered during the outbreak as a strategy for farmers to avoid losses associated with mortality.     

Survey of infectious agents involved in acute respiratory disease in finishing pigs.

Outbreaks of respiratory disease constitute a major health problem in herds of finishing pigs and their aetiology often remains unclear. In this study, 16 outbreaks of respiratory disease with acute clinical signs in finishing pigs were investigated to determine which infectious agents were involved. From each herd four diseased and two clinically healthy pigs were examined pathologically and for the presence of viruses, bacteria and mycoplasmas. In addition, paired blood samples from 10 groupmates of the diseased pigs were tested for antibodies against commonly known causal agents of respiratory disease. A clear diagnosis was possible in 12 of the 16 outbreaks. Seven were due to an infection with influenza virus and five were due to an infection with Actinobacillus pleuropneumoniae. A combination of influenza virus and A pleuropneumoniae may have caused one other outbreak, but no clear cause could be established for the other three outbreaks.     

Mechanism of thrombocytopenia in African swine fever

Pigs were inoculated with an African swine fever (ASF) isolate of moderate virulence, and the changes in the number of circulating blood platelets during infection were correlated with the appearance of antiviral antibody and fluctuations in total plasma hemolytic complement concentrations. Thrombocytopenia was detected by postinoculation days (PID) 7 and 8, and antiviral antibody was detected by PID 7, using an indirect immunofluorescence technique. The total hemolytic complement concentration was moderately and transiently decreased from PID 5 to 9, but was consistently low from PID 18 to 26. Pigs inoculated with an ASF virus isolate of greater virulence had a decrease in platelet counts on PID 6 and 7, and the total plasma hemolytic complement levels decreased in all pigs by PID 6 to 7. Antibody to ASF virus was not detected in pigs inoculated with the more virulent isolate. Pigs sensitized to ASF viral antigen with an inactivated-virus vaccine or by previous infection with ASF were challenge exposed. Sensitized pigs became clinically ill and thrombocytopenic by 24 to 72 hours earlier than did inoculated, nonsensitized pigs. Vaccinated pigs inoculated with homologous virus had lower blood virus concentrations than did nonvaccinated pigs. African swine fever virus-sensitized pigs inoculated with heterologous virus had a higher fatality rate than did nonsensitized pigs, and the pigs died peracutely, with only a few gross lesions in evidence. In vitro experiments demonstrated that ASF virus antigen induced platelet aggregation in platelet-rich plasma from recovered, nonviremic pigs. Viral antigen, antibody, or complement was not demonstrable on the surface of platelets from pigs inoculated with ASF virus isolate, by direct immunofluorescence testing.     

Retrospective genetic analysis of SAT-1 type foot-and-mouth disease outbreaks in West Africa (1975-1981)

The complete 1D genome region encoding the immunogenic and phylogenetically informative VP1 gene was genetically characterized for 23 South African Territories (SAT)-1 viruses causing foot-and-mouth (FMD) disease outbreaks in the West African region between 1975 and 1981. The results indicate that two independent outbreaks occurred, the first involved two West African countries, namely Niger and Nigeria, whilst the second affected Nigeria alone. In the former epizootic, virus circulation spanned a period of 2 years, whilst in the latter virus was recovered from the field over a 3 year period. Comparison of the West African viruses with SAT-1 viruses from other regions on the continent revealed that the two West African lineages identified in this study are regionally distinct. Furthermore, variation in VP1 gene length was identified in SAT-1 viruses for the first time, further emphasizing the uniqueness of these pathogens in West Africa. This first retrospective analysis in which the molecular epidemiology of SAT-1 viruses in West Africa is reported, provides a useful measure of the regional variation of these viruses and is an essential first step in the establishment of a West African sequence database that will be a useful reference for future outbreak eventualities.     

Coagulation changes in African swine fever virus infection

Pigs were infected with highly virulent (Tengani '62), with moderately virulent (DR '79) African swine fever (ASF) virus, or with virulent hog cholera (HC) virus. Changes in platelet counts, selected coagulation assays and concentrations of factor VIII-related antigen (VIIIR:Ag) were monitored. Permeability of aortic endothelium was studied after the injection of Evan's blue dye on various days after infection with DR '79 ASF virus. Virulent ASF virus caused prolongation of the activated partial thromboplastin time (APTT), 1-stage prothrombin time, and thrombin clotting time as early as postinoculation day (PID) 4. These changes became progressively more severe until death. Both virulent HC and DR'79 viruses induced an increase APPT and thrombin clotting time at PID 3 to 4, only occasionally did the prothrombin time increased significantly (P less than 0.01). The APPT began to decrease on PID 7 and 8, but only DR'79-infected pigs lived long enough to regain a normal APTT. Infection by ASF viruses caused acute thrombocytopenia after PID 6 and platelet counts of HC virus-infected pigs decreased progressively from the onset of fever to levels of 1 to 2 X 10(5)/mm3 at PID 6 to 7. All ASF virus-infected pigs had an increase in VIIIR:Ag beginning at PID 3, with maximum increases at PID 6 to 7. Hog cholera virus infection did not cause consistent changes in levels of VIIIR:Ag. Pigs infected with DR'79 virus did not have increased vascular permeability to Evan's blue dye during infection; however, there was markedly decreased staining of the aorta after pigs became thrombocytopenic.     

Transmissible Virus Diseases in Porcine Reproduction

Contents This paper describes the risk of transmission and possible consequences of viral diseases in pigs (as CSFV, PRRSV and SVD), transmitted via AI and embryo transfer. Transmission via A1 is, however, more unlikely for CSF and SVD than for PRRS. The likelihood of disease transmission is greater with the introduction of a boar into a herd than through the use of fresh or frozen semen. The probability that an infection with CSF or SVD virus starts within an AI centre is very small, because of the high hygienic measurements and quarantine period, although the viruses can be transmitted if these centra are located within a protection zone. Therefore, during an outbreak, it should be recommended to stop semen distribution within this zone.     

新疆乌鲁木齐市一起非洲猪瘟疫情的紧急流行病学调查

2019年4月,新疆乌鲁木齐市某生猪养殖合作社暴发非洲猪瘟疫情。为查明疫情可能来源及扩散范围,科学指导疫情处置工作,通过现场调查、问卷调查与访谈、采样检测等方式,对该合作社及流行病学相关场点开展了紧急流行病学调查。溯源调查发现,此次疫情由调运的生猪及车辆机械性带毒引发可能性最大;追踪调查结果显示,疫情局限于该养殖合作社,但对屠宰场有污染。调查结果提示,应提高养殖户生物安全意识,完善车辆洗消制度,严格调运和屠宰环节的检疫监管。     

Isolation of respiratory bovine coronavirus, other cytocidal viruses, and Pasteurella spp from cattle involved in two natural outbreaks of shipping fever

OBJECTIVE: To identify cytocidal viruses and Pasteurella spp that could be isolated from cattle involved in 2 natural outbreaks of shipping fever. ANIMALS: 105 and 120 castrated male 4- to 8-month-old feedlot cattle involved in 1997 and 1998 outbreaks, respectively. PROCEDURES: Nasal swab specimens and blood samples were collected, and cattle were vaccinated on arrival at an order-buyer barn from 4 local auction houses. Four days later, they were transported to a feedlot, and additional nasal swab specimens and blood samples were collected. Nasal swab specimens were submitted for virus isolation and bacterial culture; blood samples were submitted for measurement of respiratory bovine coronavirus (RBCV) hemagglutinin inhibition titers. RESULTS: 93 of 105 cattle and 106 of 120 cattle developed signs of respiratory tract disease during 1997 and 1998, respectively, and RBCV was isolated from 81 and 89 sick cattle, respectively, while at the order-buyer's barn or the day after arrival at the feedlot. During the 1997 outbreak, bovine herpesvirus 1 was isolated from 2 cattle at the order-buyer's barn and from 5 cattle 7 and 14 days after arrival at the feedlot, and parainfluenza virus 3 was isolated from 4 cattle 14 days after arrival at the feedlot. During the 1998 outbreak, bovine herpesvirus 1 was isolated from 2 cattle at the order-buyer's barn and on arrival at the feedlot and from 5 cattle 7 and 14 days after arrival at the feedlot, and parainfluenza virus 3 was isolated from 1 animal the day of, and from 18 cattle 7 and 14 days after, arrival at the feedlot. Pasteurella spp was cultured from 4 and 6 cattle at the order-buyer's barn and from 92 and 72 cattle on arrival at the feedlot during the 1997 and 1998 outbreaks, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that RBCV may play a causative role in outbreaks of shipping fever in cattle. More than 80% of the sick cattle shed RBCV at the beginning of 2 outbreaks when the Pasteurella spp infection rate was low.