{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T20:42:14Z","timestamp":1783716134117,"version":"3.55.0"},"reference-count":50,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,12,7]],"date-time":"2021-12-07T00:00:00Z","timestamp":1638835200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Reprod. Health"],"abstract":"<jats:p>The development of therapies targeted to improve the health of women has utilized direct vaginal delivery as a more effective and less toxic method of protection from HIV and other pathogens. Vaginal applicants and delivery devices that provide sustained effects have been met with increasing acceptability, but the efficacy and toxicity outcomes have not been successfully predicted by preclinical <jats:italic>in vitro<\/jats:italic> studies and animal modeling. We have explored the utilization of sheep as a model for testing the safety of vaginal applicants and devices based on spatial and structural similarities to the human vagina. As recently noted by the FDA, an additional safety measure is an impact on the vaginal microbiome (VMB) that is known to contribute to vaginal health and influence pathogen susceptibility and drug metabolism. To advance the utility of the sheep vaginal model, we completed a thorough molecular characterization of the ovine VMB utilizing both next-generation sequencing (NGS) and PCR methods. The process also created a custom PCR array to quantify ovine VMB community profiles in an affordable, higher throughput fashion. The results from vaginal swabs (&amp;gt;475 samples) collected from non-pregnant crossbred Dorset and Merino ewes treated with selected vaginal applicants or collected as sham samples established 16 VMB community types (VMB CTs). To associate VMB CTs with eubiosis or dysbiosis, we also completed custom ELISAs for six cytokines identifying IL1B, IL8, TNFa, and CXCL10 as useful markers to support the characterization of ovine vaginal inflammation. The results indicated that <jats:italic>Pasteurella, Actinobacillus, Pseudomonas, Bacteroides, Leptotrichia<\/jats:italic>, and <jats:italic>E. coli<\/jats:italic> were common markers of eubiosis (low inflammatory marker expression), and that <jats:italic>Haemophilus, Ureaplasma<\/jats:italic>, and <jats:italic>Corynebacterium<\/jats:italic> were associated with dysbiosis (high cytokine levels). Utilizing the optimized workflow, we also confirmed the utility of three commonly used vaginal applicants for impact on the VMB and inflammatory state, producing a dataset that supports the recommendation for the use of sheep for testing of vaginal applicants and devices as part of preclinical pipelines.<\/jats:p>","DOI":"10.3389\/frph.2021.714829","type":"journal-article","created":{"date-parts":[[2021,12,7]],"date-time":"2021-12-07T13:21:02Z","timestamp":1638883262000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Characterization of the Ovine Vaginal Microbiome and Inflammation Patterns as an Improved Testing Model of Human Vaginal Irritation"],"prefix":"10.3389","volume":"3","author":[{"given":"Richard B.","family":"Pyles","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aaron L.","family":"Miller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carrie","family":"Maxwell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lauren","family":"Dawson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicola","family":"Richardson-Harman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Glenn","family":"Swartz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cynthia","family":"O'Neill","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cattlena","family":"Walker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gregg N.","family":"Milligan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy","family":"Madsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Massoud","family":"Motamedi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gracie","family":"Vargas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen L.","family":"Vincent","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,12,7]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"324","DOI":"10.1016\/j.ajog.2018.11.1089","article-title":"Ecological dynamics of the vaginal microbiome in relation to health and disease","volume":"220","author":"Greenbaum","year":"2019","journal-title":"Am J Obstet Gynecol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/j.maturitas.2016.05.015","article-title":"Menopause and the vaginal microbiome","volume":"91","author":"Muhleisen","year":"2016","journal-title":"Maturitas."},{"key":"B3","doi-asserted-by":"publisher","first-page":"e00487","DOI":"10.1128\/IAI.00487-20","article-title":"The complex link between the female genital microbiota, genital infections, and inflammation","volume":"89","author":"Dabee","year":"2021","journal-title":"Infect Immun."},{"key":"B4","doi-asserted-by":"publisher","first-page":"3340","DOI":"10.3389\/fmicb.2018.03340","article-title":"Cultivated human vaginal microbiome communities impact zika and herpes simplex virus replication in ex vivo vaginal mucosal cultures","volume":"9","author":"Amerson-Brown","year":"2019","journal-title":"Front Microbiol."},{"key":"B5","doi-asserted-by":"publisher","first-page":"e93419","DOI":"10.1371\/journal.pone.0093419","article-title":"Cultivated vaginal microbiomes alter HIV-1 infection and antiretroviral efficacy in colonized epithelial multilayer cultures","volume":"9","author":"Pyles","year":"2014","journal-title":"PLoS ONE."},{"key":"B6","doi-asserted-by":"publisher","first-page":"e25300","DOI":"10.1002\/jia2.25300","article-title":"The genital tract and rectal microbiomes: their role in HIV susceptibility and prevention in women","volume":"22","author":"Abdool Karim","year":"2019","journal-title":"J Int AIDS Soc."},{"key":"B7","doi-asserted-by":"publisher","first-page":"103291","DOI":"10.1016\/j.jri.2021.103291","article-title":"Liebenberg L. 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