The survival of African swine fever virus (ASFV) on different matrices and its infectivity in wild as well as domestic swine is still a matter of interest. ASFV is resistant to environmental effects; this fact is enhanced by the presence of organic material. Therefore, the aim of this work was to determine the ability of laboratory ASFV to survive in soil at different temperatures (4 and 22 °C) and with and without the presence of blood using culture procedures. The suitability of the procedure for determining the viability and titre of the ASFV field strain by the hemadsorption method was also verified, when a higher decrease in virus infectivity in the case of clay compared with peat was demonstrated. The stability of the virus was clearly temperature-dependent, the infectious virus was detected after 112 days, and the viral DNA was still detected in the matrix 210 days after inoculation in a relatively high and stable concentration (between 106 and 107 genome equivalents/mL). Based on this knowledge, soil and other environmental samples could provide rapid and reliable information on the disease outbreak and serve as indicators of the risk posed by the affected locality.
African swine fever virus is the causative agent of an acute and highly contagious disease affecting domestic and wild members of the family Suidae. The virus can be transmitted by direct contact among infected animals or via a contaminated environment or feed. Since the contaminated meat or products thereof have been characterised as the most probable vehicle in several outbreaks, the aim of the present study was to define viability of the virus in meat under conditions of freezing and chilling (−25 °C and 6 °C) and low temperature cooking (55 °C for 2.5 h and for 1 h). Two independent methods were employed; cell culture as a reference and real-time polymerase chain reaction combined with palladium compound (BB-PdCl2 and PdCl2COD) pre-treatment as an alternative method. Obtained results demonstrated a minimal decrease in the infectious virus titre during storage at −25 °C, and a remaining amount of viruses in meat stored at 6 °C for 14 months that can cause a disease after ingestion. The results obtained by both methods applied on the samples corresponded to each other. In contrast, results related to the virus’ persistence in thermal-treated meat indicated much lower stability than previously thought; infectious viruses were not detected by infectivity assay after the treatment at 55 °C for 1 h. The observed difference of one order of magnitude of virus detected using palladium compound pre-treatment suggests presence of intact rather than infectious viruses. A better suitability of PdCl2COD compared to BB-PdCl2 pre-treatment was demonstrated.
African swine fever (ASF) is highly contagious haemorrhagic viral disease of domestic pigs and wild boars. The causative agent can be transmitted by direct contact with infected animals or via a contaminated environment, fomites, feed, meat and products thereof. Soft ticks (genus Ornithodoros) are known reservoirs and transmission vectors of the virus. As the disease causes serious problems in many countries, rapid detection of the agent and early diagnosis could help in prevention of its spread. Therefore, a multiple-analyte profile (xMAP) technology based on multiple oligonucleotide ligation followed by polymerase chain reaction (MOL-PCR) was introduced and verified. A system targeting two independent loci of the virus genome was designed to increase the likelihood of different strains detection and an internal control was employed to verify the correct course of the analysis. The sensitivity was experimentally determined as 10 genomic copies of the virus in one µl of isolated DNA. The system was verified on samples originating from a recent ASF outbreak in the Czech Republic (six spleen) and the Czech market (eight liver and heart tissues) with real-time polymerase chain reaction used as a reference method. The results of both methods were in agreement, even in samples with a low concentration of the virus genome (9.45 × 101 genomic copies/µl of DNA). The system introduced represents an open method allowing the detection and semi-quantification of up to 50 targets/agents in one reaction. It can, therefore, be used for rapid one-step screening and as an effective tool for risk management.
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