{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,27]],"date-time":"2026-01-27T09:27:36Z","timestamp":1769506056416,"version":"3.49.0"},"reference-count":31,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2025,4,22]],"date-time":"2025-04-22T00:00:00Z","timestamp":1745280000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100001458","name":"American Kennel Club Canine Health Foundation","doi-asserted-by":"publisher","award":["02519"],"award-info":[{"award-number":["02519"]}],"id":[{"id":"10.13039\/100001458","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the State of North Carolina"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:sec id=\"sec011\">\n<jats:title>Introduction<\/jats:title>\n<jats:p>The potential role of pathogens, particularly vector-transmitted infectious agents, as a cofactor or cause of neoplasia has not been intensively investigated. We previously reported a potential link between <jats:italic>Bartonella<\/jats:italic> spp. bacteremia and splenic hemangiosarcoma (HSA) in dogs living in the United States. The purpose of this study was to: 1\/ further determine the prevalence of <jats:italic>Bartonella<\/jats:italic> spp. DNA in dogs with splenic HSA from throughout the United States; 2\/ assess the impact of sample preservation methods on <jats:italic>Bartonella<\/jats:italic> spp. DNA amplification using characterized tissue samples from dogs diagnosed with HSA.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec012\">\n<jats:title>Methods<\/jats:title>\n<jats:p>In a blinded manner, we determined the presence of <jats:italic>Bartonella<\/jats:italic> spp. DNA in scrolls from biorepository formalin-fixed paraffin-embedded (FFPE) spleens from dogs living in three distant locations geographically transecting the United States. DNA extracted from non-lesional spleens (<jats:italic>n<\/jats:italic> = 249), nodular lymphoid hyperplasia spleens (<jats:italic>n<\/jats:italic> = 248), and splenic HSA (<jats:italic>n<\/jats:italic> = 330) were tested by quantitative polymerase chain reaction (qPCR), and droplet digital PCR (ddPCR). Subsequently, <jats:italic>Bartonella<\/jats:italic> PCR results from FFPE tissues and formalin-fixed tissues were compared using previously tested fresh frozen tissues from an additional 48 dogs with HSA.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec013\">\n<jats:title>Results<\/jats:title>\n<jats:p>There was no significant difference in the proportion of <jats:italic>Bartonella<\/jats:italic> PCR positive FFPE tissues from dogs diagnosed with an alesional spleen, nodular lymphoid hyperplasia, and splenic HSA. Regardless of the histological diagnosis, the most common <jats:italic>Bartonella<\/jats:italic> species identified was <jats:italic>B<\/jats:italic>. <jats:italic>henselae<\/jats:italic> (32\/38). <jats:italic>Bartonella<\/jats:italic> spp. DNA was detected in a significantly larger proportion of fresh frozen tissues compared to FFPE tissues, when tested by qPCR (22\/48 versus 1\/48; <jats:italic>p<\/jats:italic> &lt;0.0001) or ddPCR (19\/48 versus 1\/48; <jats:italic>p<\/jats:italic> &lt;0.0001). Using ddPCR, <jats:italic>Bartonella<\/jats:italic> DNA was more often amplified from formalin-fixed tissues compared to FFPE tissues (15\/39 versus 1\/39; <jats:italic>p<\/jats:italic> &lt;0.0001). The sensitivity of qPCR on FFPE samples and formalin-fixed samples, when comparing to fresh frozen samples as the reference standard, was 4.5% and 11.8%, respectively.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec014\">\n<jats:title>Conclusion<\/jats:title>\n<jats:p>Due to decreased DNA amplification efficiency, FFPE scrolls should not be used for the detection of <jats:italic>Bartonella<\/jats:italic> infection in spleen samples from dogs with HSA. PCR testing of fresh frozen tissues substantially improves the detection of <jats:italic>Bartonella<\/jats:italic> spp. infection. If fresh frozen tissues are not available, formalin-fixed tissues should be tested with digital PCR to enhance <jats:italic>Bartonella<\/jats:italic> DNA detection.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0321806","type":"journal-article","created":{"date-parts":[[2025,4,22]],"date-time":"2025-04-22T17:32:50Z","timestamp":1745343170000},"page":"e0321806","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Molecular detection of Bartonella spp. DNA in dogs with hemangiosarcoma"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2264-307X","authenticated-orcid":true,"given":"Cynthia","family":"Robveille","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ricardo G.","family":"Maggi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0949-5698","authenticated-orcid":true,"given":"Erin","family":"Lashnits","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5740-9550","authenticated-orcid":true,"given":"Taryn A.","family":"Donovan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keith E.","family":"Linder","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel P.","family":"Regan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin 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