6f5669add722936b8794877e0f23b1e6 pmc_11857857.pdf 4f8fe23ec0f50baed912c28395fa7c2f8e662dc4 pmc_11857857.pdf 543da53c603aaddd1a799c77d1c4bf6764a508e15e829d0d9865915fc918a26a pmc_11857857.pdf Title: Evaluation of the Dose of African Swine Fever Virus Required to Establish Infection in Pigs Following Oral Uptake Subject: African swine fever virus (ASFV) is known to be very stable within a protein-rich environment and indirect virus transmission can be mediated via oral uptake of different materials. However, experimental studies in pigs have shown that infection by ASFV via the oral route can be difficult to establish. Currently, there is a lack of studies using strict oral inoculations of pigs with different doses of ASFV. Therefore, we aimed to determine the dose of a European genotype II ASFV that is required to establish infection of pigs by the oral route. In this study, 24 pigs were divided into four groups of six. Three of the groups were fed with a low, medium or high dose of the ASFV POL/2015/Podlaskie virus. The pigs in the fourth group served as positive controls and were inoculated intranasally, just once, using the low dose of the virus. All the pigs inoculated intranasally with ASFV succumbed to the infection, while only three of the six pigs that were fed the high dose of the virus became infected. None of the 12 pigs that were fed with either the medium or low dose of the virus became infected, despite receiving up to thirteen doses each. In two of the pigs infected by intranasal inoculation, the presence of a variant form of the ASFV genome was detected. The results obtained in this study underline that ASFV infection is more difficult to establish via the oral route when compared to the intranasal route. The high dose needed in order to establish oral infection could have implications for future strategies using baited vaccines containing infectious live-attenuated ASFV. Keywords: ASFV; dosing study; feeding; transmission; oral uptake Author: Ann Sofie Olesen, Christina Marie Lazov, Francesc Accensi, Camille Melissa Johnston, Thomas Bruun Rasmussen, Anette Bøtner, Louise Lohse and Graham J. Belsham Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Sat Feb 8 13:08:43 2025 CET ModDate: Sat Feb 8 13:11:31 2025 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 15 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2268821 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- RKFMJG+VnURWPalladioL Type 1 Custom yes yes yes 10 0 YLGYUG+URWPalladioL-Roma Type 1 Custom yes yes yes 16 0 NFWCHF+URWPalladioL-Bold Type 1 Custom yes yes yes 22 0 LTDUEW+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 OVNEXW+EURM10 Type 1 Builtin yes yes yes 53 0 PDYDVH+CMSY10 Type 1 Builtin yes yes yes 61 0 JJPJDN+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 72 0 JJPKFP+TimesNewRomanPSMT TrueType WinAnsi yes yes no 78 0 JJPJEO+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 81 0 JJPJEP+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 87 0 JJPJFA+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 93 0 JJPJFB+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 96 0 JJPJFC+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 99 0 JJPJMB+PalatinoLinotype-Roman TrueType MacRoman yes yes no 102 0 JJPJDN+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 118 0 JJPJEO+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 124 0 JJPJEP+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 130 0 JJPJFA+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 136 0 JJPJFB+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 139 0 JJPJFC+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 142 0 JJPJMB+PalatinoLinotype-Roman TrueType MacRoman yes yes no 145 0 JJPJDN+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 155 0 JJPJEO+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 161 0 JJPJEP+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 167 0 JJPJFA+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 173 0 JJPJFB+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 176 0 JJPJFC+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 179 0 JJPJMB+PalatinoLinotype-Roman TrueType MacRoman yes yes no 182 0 DPLNIF+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 196 0 DPLNIG+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 202 0 DPLNII+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 205 0 DPLNJI+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 208 0 DPLNJJ+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 214 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-03-28 02:08:29 CET RepresentationInformation: pmc_11857857.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-03-27 02:09:45 CET Size: 2268821 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 415 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Pigs and Housing Destination: subsection.2.1 Item: Title: Virus and Inoculation Material Destination: subsection.2.2 Item: Title: Study Design Destination: subsection.2.3 Item: Title: Clinical Examinations and Euthanasia Destination: subsection.2.4 Item: Title: Sampling Destination: subsection.2.5 Item: Title: qPCR Analysis of EDTA-Blood and Swab Samples Destination: subsection.2.6 Item: Title: Detection of Infectious Virus in Nasal Swabs Destination: subsection.2.7 Item: Title: Anti-ASFV Antibody Detection in Serum Destination: subsection.2.8 Item: Title: Preparation of Long PCR Products from Viral DNA Destination: subsection.2.9 Item: Title: Variant Calling Destination: subsection.2.10 Item: Title: Deletion Screening by PCR Destination: subsection.2.11 Item: Title: Nanopore Sequencing Destination: subsection.2.12 Item: Title: Results Destination: section.3 Children: Item: Title: Preparation and Back-Titration of the Inoculation Material Destination: subsection.3.1 Item: Title: Course of Infection in Intranasally Inoculated Pigs Destination: subsection.3.2 Item: Title: Course of ASFV Infection in Orally Inoculated Pigs Destination: subsection.3.3 Item: Title: Variant Analysis of ASFV in the Inoculum Destination: subsection.3.4 Item: Title: Deletion Screening by PCR and Sequencing Destination: subsection.3.5 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: appendix.A. Info: Title: Evaluation of the Dose of African Swine Fever Virus Required to Establish Infection in Pigs Following Oral Uptake Author: Ann Sofie Olesen, Christina Marie Lazov, Francesc Accensi, Camille Melissa Johnston, Thomas Bruun Rasmussen, Anette Bøtner, Louise Lohse and Graham J. Belsham Subject: African swine fever virus (ASFV) is known to be very stable within a protein-rich environment and indirect virus transmission can be mediated via oral uptake of different materials. However, experimental studies in pigs have shown that infection by ASFV via the oral route can be difficult to establish. Currently, there is a lack of studies using strict oral inoculations of pigs with different doses of ASFV. Therefore, we aimed to determine the dose of a European genotype II ASFV that is required to establish infection of pigs by the oral route. In this study, 24 pigs were divided into four groups of six. Three of the groups were fed with a low, medium or high dose of the ASFV POL/2015/Podlaskie virus. The pigs in the fourth group served as positive controls and were inoculated intranasally, just once, using the low dose of the virus. All the pigs inoculated intranasally with ASFV succumbed to the infection, while only three of the six pigs that were fed the high dose of the virus became infected. None of the 12 pigs that were fed with either the medium or low dose of the virus became infected, despite receiving up to thirteen doses each. In two of the pigs infected by intranasal inoculation, the presence of a variant form of the ASFV genome was detected. The results obtained in this study underline that ASFV infection is more difficult to establish via the oral route when compared to the intranasal route. The high dose needed in order to establish oral infection could have implications for future strategies using baited vaccines containing infectious live-attenuated ASFV. 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