{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T13:04:24Z","timestamp":1791291864417,"version":"4.1.0"},"reference-count":36,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2022,7,2]],"date-time":"2022-07-02T00:00:00Z","timestamp":1656720000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016349","name":"Egg Industry Center","doi-asserted-by":"publisher","award":["2704235"],"award-info":[{"award-number":["2704235"]}],"id":[{"id":"10.13039\/100016349","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>The present study was conducted to assess the potential vector role of feedstuffs for the area spreading of avian influenza virus (AIV). Firstly, feed samples were collected from commercial poultry facilities that experienced highly pathogenic avian influenza (H5N2) in 2014\u20132015 for AIV testing by a real-time RT\u2013PCR specific for the viral matrix gene. Secondly, feed materials obtained from an AIV-negative farm were spiked with various concentrations of a low pathogenic AIV H5N2. Virus-spiked cell culture media were prepared in the same manner and used for comparison. The spiked feed and media samples were tested by a multiplex real-time RT\u2013PCR ran in a quantitative manner, either immediately or after incubation at \u221220, 4, 22, and 37 \u00b0C for 24, 48, and 72 h. Some of the feedstuffs collected from the poultry facilities or feed mills were positive for AIV RNA but negative by the virus isolation (VI) test, while all the formaldehyde-treated feedstuffs were PCR-negative. In the spiked feeds, the AIV titer was 1\u20133 logs lower than that in the corresponding media, even when tested immediately after spiking, suggesting that feed might have a negative impact on the virus or PCR detection. The half-life of AIV RNA was shorter at a higher temperature. A significant decay in the viral RNA over time was noted at 37 \u00b0C (p &lt; 0.05), suggesting that feedstuffs should be maintained in the cold chain when testing is desired. Furthermore, the thermal degradation of AIV suggests that the heat treatment of feeds could be an alternative to chemical treatment when contamination is suspected. Collectively, the study observations indicate that AIV survivability in feed is relatively low, thus rendering it a low risk.<\/jats:p>","DOI":"10.3390\/pathogens11070755","type":"journal-article","created":{"date-parts":[[2022,7,4]],"date-time":"2022-07-04T20:59:18Z","timestamp":1656968358000},"page":"755","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Evaluation of Feedstuffs as a Potential Carrier of Avian Influenza Virus between Feed Mills and Poultry Farms"],"prefix":"10.3390","volume":"11","author":[{"given":"Shahan","family":"Azeem","sequence":"first","affiliation":[{"name":"Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"},{"name":"Institute of Microbiology, Faculty of Veterinary Science, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuko","family":"Sato","sequence":"additional","affiliation":[{"name":"Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baoqing","family":"Guo","sequence":"additional","affiliation":[{"name":"Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1190-5713","authenticated-orcid":false,"given":"Anna","family":"Wolc","sequence":"additional","affiliation":[{"name":"Department of Animal Science, College of Agriculture and Life Sciences, Iowa State University, Ames, IA 50011, USA"},{"name":"Hy-Line International, Dallas Center, IA 50063, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanjun","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hai","family":"Hoang","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mahesh","family":"Bhandari","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen","family":"Mayo","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Yuan","sequence":"additional","affiliation":[{"name":"Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jihun","family":"Yoon","sequence":"additional","affiliation":[{"name":"College of Engineering, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2540-8769","authenticated-orcid":false,"given":"Phillip C.","family":"Gauger","sequence":"additional","affiliation":[{"name":"Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3969-7285","authenticated-orcid":false,"given":"Kyoung-Jin","family":"Yoon","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"},{"name":"Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,2]]},"reference":[{"key":"ref_1","unstructured":"ICTV (2022, May 25). International Committee on Taxonomy of Viruses. Orthomyxoviridae. Available online: https:\/\/talk.ictvonline.org\/ictv-reports\/ictv_9th_report\/negative-sense-rna-viruses-2011\/w\/negrna_viruses\/209\/orthomyxoviridae."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1128\/mr.56.1.152-179.1992","article-title":"Evolution and ecology of influenza A viruses","volume":"56","author":"Webster","year":"1992","journal-title":"Microbiol. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"4269","DOI":"10.1073\/pnas.1116200109","article-title":"A distinct lineage of influenza A virus from bats","volume":"109","author":"Tong","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Tong, S., Zhu, X., Li, Y., Shi, M., Zhang, J., Bourgeois, M., Yang, H., Chen, X., Recuenco, S., and Gomez, J. (2013). New World Bats Harbor Diverse Influenza A Viruses. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003657"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Munster, V.J., Baas, C., Lexmond, P., Waldenstrom, J., Wallensten, A., Fransson, T., Rimmelzwaan, G.F., Beyer, W.E., Schutten, M., and Olsen, B. (2007). Spatial, temporal, and species variation in prevalence of influenza A viruses in wild migratory birds. PLoS Pathog., 3.","DOI":"10.1371\/journal.ppat.0030061"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Global Consortium for H5N8 and Related Influenza Viruses (2016). Role for migratory wild birds in the global spread of avian influenza H5N8. Science, 354, 213\u2013216.","DOI":"10.1126\/science.aaf8852"},{"key":"ref_7","unstructured":"United Stated Department of Agriculture-Animal and Plant Health Inspection Agency-Veterinary Services (2022, May 25). 2022 Detections of Highly Pathogenic Avian Influenza in Wild Birds, Available online: https:\/\/www.aphis.usda.gov\/aphis\/ourfocus\/animalhealth\/animal-disease-information\/avian\/avian-influenza\/hpai-2022\/2022-hpai-wild-birds."},{"key":"ref_8","unstructured":"United Stated Department of Agriculture-Animal and Plant Health Inspection Agency (2022, May 25). USDA Confirms Highly Pathogenic H7N3 Avian Influenza in a Commercial Flock in Chesterfield County, South Carolina, Available online: https:\/\/www.aphis.usda.gov\/aphis\/newsroom\/stakeholder-info\/sa_by_date\/sa-2020\/sa-04\/hpai-sc."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.3201\/eid\/1706.100880","article-title":"Wild birds and increased transmission of highly pathogenic avian influenza (H5N1) among poultry, Thailand","volume":"17","author":"Keawcharoen","year":"2011","journal-title":"Emerg. Infect. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1637\/11137-050815-Reg","article-title":"High Rates of Detection of Clade 2.3.4.4 Highly Pathogenic Avian Influenza H5 Viruses in Wild Birds in the Pacific Northwest During the Winter of 2014\u201315","volume":"60","author":"Ip","year":"2016","journal-title":"Avian Dis."},{"key":"ref_11","unstructured":"Centre for Infectious Disease Research and Policy (2022, May 25). Report Finds $1.2 Billion in Iowa Avian Flu Damage. Available online: http:\/\/www.cidrap.umn.edu\/news-perspective\/2015\/08\/report-finds-12-billion-iowa-avian-flu-damage."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"340","DOI":"10.3201\/eid2302.161963","article-title":"H5Nx Panzootic Bird Flu\u2014Influenza\u2019s Newest Worldwide Evolutionary Tour","volume":"23","author":"Taubenberger","year":"2017","journal-title":"Emerg. Infect. Dis."},{"key":"ref_13","unstructured":"United Stated Department of Agriculture-Animal and Plant Health Inspection Agency-Veterinary Services (2022, May 25). 2016 HPAI Preparedness and Response Plan, Available online: https:\/\/www.aphis.usda.gov\/animal_health\/downloads\/animal_diseases\/ai\/hpai-preparedness-and-response-plan-2015.pdf."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"886","DOI":"10.3201\/eid2105.142020","article-title":"Novel Eurasian Highly Pathogenic Avian Influenza a H5 Viruses in Wild Birds, Washington, USA, 2014","volume":"21","author":"Ip","year":"2015","journal-title":"Emerg. Infect. Dis."},{"key":"ref_15","unstructured":"United Stated Department of Agriculture-Animal and Plant Health Inspection Agency-Veterinary Services (2022, May 25). Epidemiologic and Other Analyses of HPAI-Affected Poultry Flocks: June 15, 2015 Report, Available online: http:\/\/www.aphis.usda.gov\/animal_health\/animal_dis_spec\/poultry\/downloads\/Epidemiologic-Analysis-June-15-2015.pdf."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Spackman, E., Pantin-Jackwood, M.J., Kapczynski, D.R., Swayne, D.E., and Suarez, D.L. (2016). H5N2 Highly pathogenic avian influenza viruses from the US 2014\u20132015 outbreak have an unusually long pre-clinical period in turkeys. BMC Vet. Res., 12.","DOI":"10.1186\/s12917-016-0890-6"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Swayne, D.E. (2008). High pathogenicity avian influenza in the Americas. Avian Influenza, Blackwell Publishing.","DOI":"10.1002\/9780813818634"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1637\/11465-070616-ResNote","article-title":"Inactivation of avian influenza virus in nonpelleted chicken feed","volume":"60","author":"Toro","year":"2016","journal-title":"Avian Dis."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Spackman, E. (2008). Avian influenza virus isolation and propagation in chicken eggs. Avian Influenza Virus, Humana Press.","DOI":"10.1007\/978-1-59745-279-3"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Kawaoka, Y., and Neumann, G. (2012). Infleunza virus isolation. Influenza Virus: Methods and Protocols, Methods in Molecular Biology, Humana Press.","DOI":"10.1007\/978-1-61779-621-0"},{"key":"ref_21","unstructured":"Dufour-Zavala, L., Swayne, D.E., Glisson, J.R., Pearson, J.E., Reed, W.M., and Jackwood, M.W. (2008). Virus propagation in embryonating eggs. A Laboratory Manual for the Isolation, Identification and Characterization of Avian Pathogens, American Association of Avian Pathologists. [5th ed.]."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Spackman, E. (2008). Hemagglutination assay for the avian influenza virus. Avian Influenza Virus, Humana Press.","DOI":"10.1007\/978-1-59745-279-3"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Spackman, E. (2014). Avian influenza virus detection and quantitation by real-time RT-PCR. Animal Influenza Virus, Humana Press.","DOI":"10.1007\/978-1-4939-0758-8_10"},{"key":"ref_24","unstructured":"R Core Team (2017, July 03). The R Project for Statistical Computing (ver 3.3.3). Available online: http:\/\/www.r-project.org\/."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1475-2867-2-1","article-title":"Real-time RT-PCR analysis of mRNA decay: Half-life of beta-actin mRNA in human leukemia CCRF-CEM and Nalm-6 cell lines","volume":"2","author":"Leclerc","year":"2002","journal-title":"Cancer Cell Int."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1007\/BF00347743","article-title":"The use of half-lives and associated confidence intervals in biological research","volume":"14","author":"Bryan","year":"1990","journal-title":"Vet. Res. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1637\/8736-032509-ResNote.1","article-title":"Avian Influenza Virus H9N2 Survival at Different Temperatures and pHs","volume":"54","author":"Davidson","year":"2010","journal-title":"Avian Dis."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1186\/1743-422X-6-38","article-title":"Avian influenza virus (H5N1); effects of physico-chemical factors on its survival","volume":"6","author":"Shahid","year":"2009","journal-title":"Virol. J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"131","DOI":"10.3389\/fevo.2018.00131","article-title":"Environmental persistence of influenza H5N1 is driven by temperature and salinity: Insights from a Bayesian meta-analysis","volume":"6","author":"Martin","year":"2018","journal-title":"Front. Ecol. Evol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Dee, N., Havas, K., Shah, A., Singrey, A., Spronk, G., Niederwerder, M., Nelson, E., and Dee, S. (2022). Evaluating the effect of temperature on viral survival in plant-based feed during storage. Transbound. Emerg. Dis., 1\u20136.","DOI":"10.1111\/tbed.14546"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1637\/8229-012508-Reg.1","article-title":"Using mean infectious dose of high- and low-pathogenicity avian influenza viruses originating from wild duck and poultry as one measure of infectivity and adaptation to poultry","volume":"52","author":"Swayne","year":"2008","journal-title":"Avian Dis."},{"key":"ref_32","unstructured":"United Stated Department of Agriculture-Animal and Plant Health Inspection Agency-Veterinary Services (2022, May 25). Risk That Poultry Feed Made with Corn\u2014Potentially Contaminated with Eurasian-North American Lineage H5N2 HPAI Virus from Wild Migratory Birds\u2014Results in Exposure of Susceptible Commercial Poultry, Available online: https:\/\/www.aphis.usda.gov\/animal_health\/animal_dis_spec\/poultry\/downloads\/hpai_contaminated_feed.pdf."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1186\/s40813-017-0064-3","article-title":"Survival of porcine epidemic diarrhea virus (PEDV) in thermally treated feed ingredients and on surfaces","volume":"3","author":"Trudeau","year":"2017","journal-title":"Porcine Health Manag."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"891","DOI":"10.3201\/eid2505.181495","article-title":"Infectious dose of African swine fever virus when consumed naturally in liquid or feed","volume":"25","author":"Niederwerder","year":"2019","journal-title":"Emerg. Infect. Dis."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1623","DOI":"10.1111\/tbed.13498","article-title":"Stability of classical swine fever virus and pseudorabies virus in animal feed ingredients exposed to transpacific shipping conditions","volume":"67","author":"Stoian","year":"2020","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Dee, S.A., Bauermann, F.V., Niederwerder, M.C., Singrey, A., Clement, T., and de Lima, M. (2018). Survival of viral pathogens in animal feed ingredients under transboundary shipping models. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0194509"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/11\/7\/755\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:42:09Z","timestamp":1760139729000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/11\/7\/755"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,2]]},"references-count":36,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["pathogens11070755"],"URL":"https:\/\/doi.org\/10.3390\/pathogens11070755","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,2]]}}}