Methods for Growing and Titrating African Swine Fever Virus: Field and Laboratory Samples

Carrascosa, Ángel L.; Bustos, María José; León, Patricia de

doi:10.1002/0471143030.cb2614s53

Cited by 111 publications

(108 citation statements)

References 17 publications

(37 reference statements)

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“…For detection of viable ASFV in serum and tissue samples, the haemadsorption test (HAT) was employed as previously described (Pietschmann et al., ) on the basis of standard procedures (Carrascosa, Bustos, & de Leon, ). In brief, 100 μl of sera/tissue samples was added in duplicates to 200 μl PBMC‐derived macrophages seeded in 48‐well microplates (5 × 10 6 cells/μl) which were allowed to mature for 3 days.…”

Section: Methodsmentioning

confidence: 99%

No evidence for long-term carrier status of pigs after African swine fever virus infection

Petrov

Forth

Zani

et al. 2018

Transbound Emerg Dis

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This study targeted the assessment of a potential African swine fever virus (ASFV) carrier state of 30 pigs in total which were allowed to recover from infection with ASFV "Netherlands'86" prior exposure to six healthy sentinel pigs for more than 2 months. Throughout the whole trial, blood and swab samples were subjected to routine virological and serological investigations. At the end of the trial, necropsy of all animals was performed and viral persistence and distribution were assessed. Upon infection, a wide range of clinical and pathomorphological signs were observed. After an initial acute phase in all experimentally inoculated pigs, 66.6% recovered completely and seroconverted. However, viral genome was detectable in blood samples for up to 91 days. Lethal outcomes were observed in 33.3% of the pigs with both acute and prolonged courses. No ASFV transmission occurred over the whole in-contact phase from survivors to sentinels. Similarly, infectious ASFV was not detected in any of the tissue samples from ASFV convalescent and in-contact pigs. These findings indicate that the suggested role of ASFV survivors is overestimated and has to be reconsidered thoroughly for future risk assessments.

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Section: Methodsmentioning

confidence: 99%

No evidence for long-term carrier status of pigs after African swine fever virus infection

Petrov

Forth

Zani

et al. 2018

Transbound Emerg Dis

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“…For the experimental in vivo studies, three ASFV isolates belonging to p72 genotype II obtained from the outbreaks in Armenia in 2007 (Arm07), Ukraine in 2012 (Ukr12/ Zapo), and Lithuania in 2014 (LT14/1490) were used. The ASFV isolates were propagated (2 to 6 passages) in porcine blood monocytes (PBM) recovered from naive domestic pigs (43,44). Titrations of ASFV stocks were performed using a hemadsorption assay to monitor the endpoint dilution of ASFV isolates into PBM.…”

Section: Methodsmentioning

confidence: 99%

Assessment of African Swine Fever Diagnostic Techniques as a Response to the Epidemic Outbreaks in Eastern European Union Countries: How To Improve Surveillance and Control Programs

et al. 2015

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A frican swine fever (ASF) is a complex and lethal viral disease affecting swine and has a significant socioeconomic impact on both the developed and developing world. It has a major negative effect on national, regional, and international trade and constrains pig production in affected areas. The devastating acute form of the disease is characterized, among other features, by functional and congestive-hemorrhagic disorders of the digestive and respiratory systems and causes around 100% mortality in infected pigs (1). Both European wild boars (Sus scrofa) and feral pigs are susceptible and exhibit clinical signs and mortality rates similar to those of domestic pigs. In contrast, African wild pigs (Phacochoerus and Potamochoerus spp.) are resistant to the disease (2-10).The causative agent of the disease, the ASF virus (ASFV), is a large double-stranded DNA virus and the only member of the Asfarviridae family, genus Asfivirus (11, 12). The virus genome is 170 to 192 kb long (13-17). ASF is endemic in sub-Saharan Africa, where it was first described in 1921 (18). Several outbreaks have occurred since then in Europe and South and Central America. In most non-African countries, the disease has been successfully eradicated, the only exception being Sardinia (Italy), where the disease is still endemic (19,20). In April 2007, the disease spread from East Africa to the Republic of Georgia (21), and outbreaks occurred in Armenia, Azerbaijan, and the Russian Federation (22). The ongoing spread of ASFV into adjacent eastern European countries, such as Ukraine (23,24) and Belarus (25), and the situation in Russia affecting both wild boars and domestic pigs placed neighboring areas in the European Union (EU) at risk for the spread of ASFV. The first cases of ASF in wild boars in Lithuania and Poland were reported in early 2014 in areas bordering . According to the World Organisation for Animal Health (OIE), during 2014, nearly 260 ASF cases or outbreaks in wild boars and domestic pigs were detected in EU countries (Latvia, Lithuania, Estonia, and Poland). This situation, combined with the uncertainty present in Belarus, has created a permanent risk of reintroducing ASF into the EU via wild boars or the illegal trade of contaminated pork products and waste (31).No vaccine is available to prevent ASF infection. The control and eradication measures applicable are based on classical disease control methods, including surveillance, epidemiological investigation, tracing of pigs, and stamping out in infected holdings. Citation Gallardo C, Nieto R, Soler A, Pelayo V, Fernández-Pinero J, MarkowskaDaniel I, Pridotkas G, Nurmoja I, Granta R, Simón A, Pérez C, Martín E, Fernández-Pacheco P, Arias M. 2015. Assessment of African swine fever diagnostic techniques as a response to the epidemic outbreaks in eastern European Union countries: how to improve surveillance and control programs.

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“…Additionally, the animal was found to have a high antibody titre (1:327,680) to ASFV by the indirect immunoperoxidase test (IPT) (Gallardo et al., ). The serum sample was subjected to molecular characterization (Bastos et al., ), virus isolation (VI) and haemadsorption (HAD) test in primary swine blood monocyte cell cultures (PBM) (Carrascosa, Bustos, & de Leon, ). The virus, named Lv17/WB/Rie1, was able to replicate in PBM in the absence of HAD as it was verified in three subsequent passages by PCR (Fernandez‐Pinero et al., ).…”

Section: The Studymentioning

confidence: 99%

“…Twenty‐one different tissues were obtained at necropsy (Table ). ASFV detection was performed in tissues and EDTA‐blood by PCR and VI (Carrascosa et al., ; Fernandez‐Pinero et al., ). The ASFV antibody titres were determined by end‐point dilution using IPT (Gallardo et al., ).…”

Section: The Studymentioning

confidence: 99%

Attenuated and non‐haemadsorbing (non‐HAD) genotypeIIAfrican swine fever virus (ASFV) isolated in Europe, Latvia 2017

Gallardo

Soler

Rodze

et al. 2019

Transbound Emerg Dis

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121

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A non‐haemadsorbing (non‐HAD) ASF virus (ASFV) genotype II, namely Lv17/WB/Rie1, was isolated from a hunted wild boar in Latvia in 2017. Domestic pigs experimentally infected with the non‐HAD ASFV developed a nonspecific or subclinical form of the disease. Two months later, these animals were fully protected when exposed to other domestic pigs infected with a related virulent HAD genotype II ASFV.

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Methods for Growing and Titrating African Swine Fever Virus: Field and Laboratory Samples

Cited by 111 publications

References 17 publications

No evidence for long-term carrier status of pigs after African swine fever virus infection

No evidence for long-term carrier status of pigs after African swine fever virus infection

Assessment of African Swine Fever Diagnostic Techniques as a Response to the Epidemic Outbreaks in Eastern European Union Countries: How To Improve Surveillance and Control Programs

Attenuated and non‐haemadsorbing (non‐HAD) genotypeIIAfrican swine fever virus (ASFV) isolated in Europe, Latvia 2017

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