{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T16:36:21Z","timestamp":1789144581970,"version":"build-2803163510"},"reference-count":56,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,8,6]],"date-time":"2025-08-06T00:00:00Z","timestamp":1754438400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"United States Department of Agriculture (USDA) Animal and Plant Health Inspection Service (APHIS)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>African swine fever (ASF) needs to be controlled, and prevention of the spread of African swine fever virus (ASFV) is dependent on enhanced surveillance and early disease detection. Commercial swine operations, especially in North America, Europe, and Asia, are characterized by comparatively large numbers of pigs, and sampling individual pigs, which represents the main strategy for current ASF surveillance, can be both costly and labor intensive. A study performed in Ghana was designed to estimate the diagnostic sensitivity of pen-based aggregate oral fluid testing for ASFV in infected pigs in a pen of 30 animals and to evaluate its utility as a tool to support surveillance of ASF in the US. This study was performed in three phases: (i) virus (Ghana ASFV24) amplification in a target host species to generate the challenge inoculum; (ii) titration of the inoculum (10% spleen homogenate) in target host species to determine the minimum dose inducing acute ASF in pigs with survival up to 5\u20136 days post-inoculation (dpi); and (iii) the main study, involving 186 pigs, consisting of 6 replicates of 30 pigs per pen and one seeder pig inoculated with wildtype ASFV (highly virulent genotype II) per pen. Daily sampling of aggregate oral fluids, uncoagulated blood, oropharyngeal swabs, fecal and water nipple swabs, and recording of rectal temperatures and clinical observations was carried out. The seeder pigs were each inoculated intramuscularly with 0.5 mL of the 10% spleen homogenate, which induced the desired clinical course of ASF in the pigs, with survival of up to 6 dpi. ASFV DNA was detected in the seeder pigs as early as 1 dpi and 2 dpi in the blood and oropharyngeal swabs, respectively. Transmission of ASFV from the seeder pigs to the contact pig population was detected via positive amplification of ASFV DNA in aggregate oral fluid samples at 3 days post-contact (dpc) in 4 out of 6 pens, and in all 6 pens, at 4 dpc. Testing of oropharyngeal swabs and blood samples from individual pigs revealed a variable number of ASFV-positive pigs between 3 and 5 dpc, with detection of 100% positivity between 6 and 18 dpc, the study endpoint. These findings demonstrate the potential utility of aggregate oral fluid sampling for sensitive and early detection of ASFV incursion into na\u00efve swine herds. It also demonstrates that testing of environmental samples from the premises could further enhance overall ASF early detection and surveillance strategies.<\/jats:p>","DOI":"10.3390\/v17081089","type":"journal-article","created":{"date-parts":[[2025,8,6]],"date-time":"2025-08-06T15:09:53Z","timestamp":1754492993000},"page":"1089","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Evaluation of Aggregate Oral Fluid Sampling for Early Detection of African Swine Fever Virus Infection"],"prefix":"10.3390","volume":"17","author":[{"given":"Bonto","family":"Faburay","sequence":"first","affiliation":[{"name":"Foreign Animal Disease Diagnostic Laboratory, National Bio and Agro-Defense Facility, Animal and Plant Health Inspection Service, United States Department of Agriculture, Manhattan, KS 66502, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen","family":"O\u2019Hara","sequence":"additional","affiliation":[{"name":"Center for Epidemiology and Animal Health, Strategy and Policy, Animal and Plant Health Inspection Service, Veterinary Services, United States Department of Agriculture, Fort Collins, CO 80523, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-6518-7068","authenticated-orcid":false,"given":"Marta","family":"Remmenga","sequence":"additional","affiliation":[{"name":"Center for Epidemiology and Animal Health, Strategy and Policy, Animal and Plant Health Inspection Service, Veterinary Services, United States Department of Agriculture, Fort Collins, CO 80523, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7092-7237","authenticated-orcid":false,"given":"Theophilus","family":"Odoom","sequence":"additional","affiliation":[{"name":"Veterinary Services Department, Ministry of Food and Agriculture, Accra M-37, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0829-5774","authenticated-orcid":false,"given":"Sherry","family":"Johnson","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine, University of Ghana, Accra LG-25, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2125-6382","authenticated-orcid":false,"given":"William","family":"Tasiame","sequence":"additional","affiliation":[{"name":"School of Veterinary Medicine, Kwame Nkrumah University of Science and Technology, Kumasi 00233, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4435-3206","authenticated-orcid":false,"given":"Matilda","family":"Ayim-Akonor","sequence":"additional","affiliation":[{"name":"CSIR-Animal Research Institute, Accra P.O. Box AH 20, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-1509-691X","authenticated-orcid":false,"given":"Benita","family":"Anderson","sequence":"additional","affiliation":[{"name":"Veterinary Services Department, Ministry of Food and Agriculture, Accra M-37, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-7398-1732","authenticated-orcid":false,"given":"Kingsley","family":"Kwabena Amoako","sequence":"additional","affiliation":[{"name":"Veterinary Services Department, Ministry of Food and Agriculture, Accra M-37, Ghana"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diane","family":"Holder","sequence":"additional","affiliation":[{"name":"Foreign Animal Disease Diagnostic Laboratory, National Bio and Agro-Defense Facility, Animal and Plant Health Inspection Service, United States Department of Agriculture, Manhattan, KS 66502, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3475-6335","authenticated-orcid":false,"given":"Wu","family":"Ping","sequence":"additional","affiliation":[{"name":"Foreign Animal Disease Diagnostic Laboratory (FADDL), National Veterinary Services Laboratories, Animal and Plant Health Inspection Service, United State Department of Agriculture, Plum Island Animal Disease Center, Plum Island, Orient, NY 11957, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michelle","family":"Zajac","sequence":"additional","affiliation":[{"name":"Foreign Animal Disease Diagnostic Laboratory, National Bio and Agro-Defense Facility, Animal and Plant Health Inspection Service, United States Department of Agriculture, Manhattan, KS 66502, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7169-3954","authenticated-orcid":false,"given":"Vivian","family":"O\u2019Donnell","sequence":"additional","affiliation":[{"name":"Foreign Animal Disease Diagnostic Laboratory (FADDL), National Veterinary Services Laboratories, Animal and Plant Health Inspection Service, United State Department of Agriculture, Plum Island Animal Disease Center, Plum Island, Orient, NY 11957, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-6002-9007","authenticated-orcid":false,"given":"Lizhe","family":"Xu","sequence":"additional","affiliation":[{"name":"Foreign Animal Disease Diagnostic Laboratory (FADDL), National Veterinary Services Laboratories, Animal and Plant Health Inspection Service, United State Department of Agriculture, Plum Island Animal Disease Center, Plum Island, Orient, NY 11957, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robin","family":"Holland","sequence":"additional","affiliation":[{"name":"Foreign Animal Disease Diagnostic Laboratory, National Bio and Agro-Defense Facility, Animal and Plant Health Inspection Service, United States Department of Agriculture, Manhattan, KS 66502, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0332-6159","authenticated-orcid":false,"given":"Corrie","family":"Brown","sequence":"additional","affiliation":[{"name":"LifeStock International, Athens, GA 30606, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Randall","family":"Levings","sequence":"additional","affiliation":[{"name":"National Veterinary Services Laboratories, Animal and Plant Health Inspection Service, Veterinary Services, United States Department of Agriculture, Ames, IA 50010, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3794-3040","authenticated-orcid":false,"given":"Suelee","family":"Robbe-Austerman","sequence":"additional","affiliation":[{"name":"National Veterinary Services Laboratories, Animal and Plant Health Inspection Service, Veterinary Services, United States Department of Agriculture, Ames, IA 50010, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1136\/vr.h699","article-title":"African swine fever in wild boar in Europe: A notable challenge","volume":"176","author":"Neimanis","year":"2015","journal-title":"Vet. 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