5bef13ccdaf3a8af25eb2b78ed358bdd pmc_9864530.pdf ff33bcbe23cf5625109ec1a65a7e8403b5d7e5c8 pmc_9864530.pdf 761e99f1d1a49701d49a9d427825b05833ac8242d1e5f08cf544a8860e585866 pmc_9864530.pdf Title: Uptake and Survival of African Swine Fever Virus in Mealworm (Tenebrio molitor) and Black Soldier Fly (Hermetia illucens) Larvae Subject: Insect production offers a sustainable source of nutrients for livestock. This comes with a risk for transmission of pathogens from the insects into the livestock sector, including viruses causing serious diseases, such as African swine fever virus (ASFV), classical swine fever virus and foot-and-mouth disease virus. ASFV is known to survive for a long time within animal meat and byproducts. Therefore, we conducted experimental exposure studies of insects to ASFV using larvae of two key insect species produced for food and feed, the mealworm; Tenebrio molitor, and the black soldier fly, Hermetia illucens. The larvae were exposed to ASFV POL/2015/Podlaskie, via oral uptake of serum or spleen material from ASFV-infected pigs. Using qPCR, the amounts of viral DNA present immediately after exposure varied from ~104.7 to 107.2 genome copies per insect. ASFV DNA was detectable in the larvae of H. illucens for up to 3 days post exposure and in T. molitor larvae for up to 9 days post exposure. To assess the presence of infectious virus within the larvae and with this, the risk of virus transmission via oral consumption, pigs were fed cakes containing larvae exposed to ASFV. Pigs that consumed 50 T. molitor or 50 H. illucens virus-exposed larvae did not become infected with ASFV. Thus, it appears, that in our experimental setting, the risk of ASFV transmission via consumption of unprocessed insect larvae, used as feed, is low. Keywords: African swine fever virus; black soldier fly; feed safety; insect rearing; mealworm; virus survival; virus transmission Author: Ann Sofie Olesen, Christina Marie Lazov, Antoine Lecocq, Francesc Accensi, Annette Bruun Jensen, Louise Lohse, Thomas Bruun Rasmussen, Graham J. Belsham and Anette Bøtner Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Dec 28 05:40:57 2022 CET ModDate: Wed Dec 28 06:24:49 2022 CET 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: 1665779 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- GFKJOY+VnURWPalladioL Type 1 Custom yes yes no 10 0 EPYIDY+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 HBSZFV+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 KTWNQB+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 HSYNCK+URWPalladioL-BoldItal Type 1 Custom yes yes no 28 0 UXHOAG+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 47 0 QMDDXF+CMSY10 Type 1 Builtin yes yes no 69 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 76 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 97 0 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TimesNewRomanPSMT CID TrueType Identity-H yes no yes 203 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-02-02 02:36:39 CET RepresentationInformation: pmc_9864530.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-01-26 02:11:44 CET Size: 1665779 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 412 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4923ab24 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Children: Item: Title: Insect Exposures Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Exposure Viruses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Experimental Insects Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Feeding of Insects Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Sampling and Euthanasia Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Pre-Processing of Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: ASFV DNA Detection by Quantitative Real-Time Polymerase Chain Reaction (qPCR) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Virus Detection by Virus Isolation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Pig Exposures Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Children: Item: Title: Pigs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Challenge Material Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Study Design Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Clinical Examination and Euthanasia Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Pig Sampling Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: ASFV DNA detection by quantitative real-time polymerase chain reaction (qPCR) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Antibody Detection Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Descriptive Statistics Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Children: Item: Title: Detection of ASFV in T. molitor Larvae and Serum Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Detection of ASFV in H. illucens Larvae, Spiked Feed Samples and Spleen Suspensions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Oral Exposure of Pigs to ASFV-Exposed T. molitor and H. illucens Larvae Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Info: Title: Uptake and Survival of African Swine Fever Virus in Mealworm (Tenebrio molitor) and Black Soldier Fly (Hermetia illucens) Larvae Author: Ann Sofie Olesen, Christina Marie Lazov, Antoine Lecocq, Francesc Accensi, Annette Bruun Jensen, Louise Lohse, Thomas Bruun Rasmussen, Graham J. Belsham and Anette Bøtner Subject: Insect production offers a sustainable source of nutrients for livestock. This comes with a risk for transmission of pathogens from the insects into the livestock sector, including viruses causing serious diseases, such as African swine fever virus (ASFV), classical swine fever virus and foot-and-mouth disease virus. ASFV is known to survive for a long time within animal meat and byproducts. Therefore, we conducted experimental exposure studies of insects to ASFV using larvae of two key insect species produced for food and feed, the mealworm; Tenebrio molitor, and the black soldier fly, Hermetia illucens. The larvae were exposed to ASFV POL/2015/Podlaskie, via oral uptake of serum or spleen material from ASFV-infected pigs. Using qPCR, the amounts of viral DNA present immediately after exposure varied from ~104.7 to 107.2 genome copies per insect. ASFV DNA was detectable in the larvae of H. illucens for up to 3 days post exposure and in T. molitor larvae for up to 9 days post exposure. To assess the presence of infectious virus within the larvae and with this, the risk of virus transmission via oral consumption, pigs were fed cakes containing larvae exposed to ASFV. Pigs that consumed 50 T. molitor or 50 H. illucens virus-exposed larvae did not become infected with ASFV. Thus, it appears, that in our experimental setting, the risk of ASFV transmission via consumption of unprocessed insect larvae, used as feed, is low. 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