{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T17:29:07Z","timestamp":1789234147205,"version":"build-2803163510"},"reference-count":39,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T00:00:00Z","timestamp":1645574400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["K22-CA226047"],"award-info":[{"award-number":["K22-CA226047"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["T32-GM007270"],"award-info":[{"award-number":["T32-GM007270"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["T32-HG000035"],"award-info":[{"award-number":["T32-HG000035"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["1849227"],"award-info":[{"award-number":["1849227"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["1701480"],"award-info":[{"award-number":["1701480"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["1950443"],"award-info":[{"award-number":["1950443"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Many pathogens can cause cancer, but cancer itself does not normally act as an infectious agent. However, transmissible cancers have been found in a few cases in nature: in Tasmanian devils, dogs, and several bivalve species. The transmissible cancers in dogs and devils are known to spread through direct physical contact, but the exact route of transmission of bivalve transmissible neoplasia (BTN) has not yet been confirmed. It has been hypothesized that cancer cells from bivalves could be released by diseased animals and spread through the water column to infect\/engraft into other animals. To test the feasibility of this proposed mechanism of transmission, we tested the ability of BTN cells from the soft-shell clam (Mya arenaria BTN, or MarBTN) to survive in artificial seawater. We found that MarBTN cells are highly sensitive to salinity, with acute toxicity at salinity levels lower than those found in the native marine environment. BTN cells also survive longer at lower temperatures, with 50% of cells surviving greater than 12 days in seawater at 10 \u00b0C, and more than 19 days at 4 \u00b0C. With one clam donor, living cells were observed for more than eight weeks at 4 \u00b0C. We also used qPCR of environmental DNA (eDNA) to detect the presence of MarBTN-specific DNA in the environment. We observed release of MarBTN-specific DNA into the water of laboratory aquaria containing highly MarBTN-diseased clams, and we detected MarBTN-specific DNA in seawater samples collected from MarBTN-endemic areas in Maine, although the copy numbers detected in environmental samples were much lower than those found in aquaria. Overall, these data show that MarBTN cells can survive well in seawater, and they are released into the water by diseased animals. These findings support the hypothesis that BTN is spread from animal-to-animal by free cells through seawater.<\/jats:p>","DOI":"10.3390\/pathogens11030283","type":"journal-article","created":{"date-parts":[[2022,2,24]],"date-time":"2022-02-24T00:53:26Z","timestamp":1645664006000},"page":"283","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":31,"title":["Survival and Detection of Bivalve Transmissible Neoplasia from the Soft-Shell Clam Mya arenaria (MarBTN) in Seawater"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7256-6490","authenticated-orcid":false,"given":"Rachael M.","family":"Giersch","sequence":"first","affiliation":[{"name":"Pacific Northwest Research Institute, Seattle, WA 98122, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5068-2199","authenticated-orcid":false,"given":"Samuel F. M.","family":"Hart","sequence":"additional","affiliation":[{"name":"Pacific Northwest Research Institute, Seattle, WA 98122, USA"},{"name":"Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satyatejas G.","family":"Reddy","sequence":"additional","affiliation":[{"name":"Bigelow Laboratory for Ocean Sciences, East Boothbay, ME 04544, USA"},{"name":"University of Georgia, Athens, GA 30602, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marisa A.","family":"Yonemitsu","sequence":"additional","affiliation":[{"name":"Pacific Northwest Research Institute, Seattle, WA 98122, USA"},{"name":"Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda J.","family":"Orellana Rosales","sequence":"additional","affiliation":[{"name":"Bigelow Laboratory for Ocean Sciences, East Boothbay, ME 04544, USA"},{"name":"Southern Maine Community College, South Portland, ME 04106, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madelyn","family":"Korn","sequence":"additional","affiliation":[{"name":"Pacific Northwest Research Institute, Seattle, WA 98122, USA"},{"name":"Tulane University, New Orleans, LA 70118, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brook M.","family":"Geleta","sequence":"additional","affiliation":[{"name":"Pacific Northwest Research Institute, Seattle, WA 98122, USA"},{"name":"Macalester College, Saint Paul, MN 55105, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter D.","family":"Countway","sequence":"additional","affiliation":[{"name":"Bigelow Laboratory for Ocean Sciences, East Boothbay, ME 04544, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7544-6918","authenticated-orcid":false,"given":"Jos\u00e9 A.","family":"Fern\u00e1ndez Robledo","sequence":"additional","affiliation":[{"name":"Bigelow Laboratory for Ocean Sciences, East Boothbay, ME 04544, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7855-1388","authenticated-orcid":false,"given":"Michael J.","family":"Metzger","sequence":"additional","affiliation":[{"name":"Pacific Northwest Research Institute, Seattle, WA 98122, USA"},{"name":"Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1016\/j.cell.2006.05.051","article-title":"Clonal origin and evolution of a transmissible cancer","volume":"126","author":"Murgia","year":"2006","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2340","DOI":"10.1111\/j.1558-5646.2009.00724.x","article-title":"Origins and evolution of a transmissible cancer","volume":"63","author":"Rebbeck","year":"2009","journal-title":"Evol. Int. J. Org. Evol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1038\/439549a","article-title":"Allograft theory: Transmission of devil facial-tumour disease","volume":"439","author":"Pearse","year":"2006","journal-title":"Nature"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1016\/j.cell.2011.11.065","article-title":"Genome sequencing and analysis of the Tasmanian devil and its transmissible cancer","volume":"148","author":"Murchison","year":"2012","journal-title":"Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/j.cell.2015.02.042","article-title":"Horizontal transmission of clonal cancer cells causes leukemia in soft-shell clams","volume":"161","author":"Metzger","year":"2015","journal-title":"Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1051\/alr:2004052","article-title":"Neoplastic diseases of commercially important marine bivalves","volume":"17","author":"Barber","year":"2004","journal-title":"Aquat. Living Resour."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.jip.2015.06.004","article-title":"Neoplastic diseases of marine bivalves","volume":"131","author":"Carballal","year":"2015","journal-title":"J. Invertebr. Pathol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.jip.2009.06.009","article-title":"Reverse transcriptase activity in tissues of the soft shell clam Mya arenaria affected with haemic neoplasia","volume":"102","author":"AboElkhair","year":"2009","journal-title":"J. Invertebr. Pathol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/0022-2011(81)90033-1","article-title":"Isolation of a viral agent causing hematopoietic neoplasia in the soft-shell clam, Mya arenaria","volume":"38","author":"Oprandy","year":"1981","journal-title":"J. Invertebr. Pathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"111","DOI":"10.3354\/dao007111","article-title":"Environmental limits for survival of sarcoma cells from the soft-shell clam Mya arenaria","volume":"7","author":"Sunila","year":"1989","journal-title":"Dis. Aquat. Org."},{"key":"ref_11","unstructured":"Moore, J.D. (1993). Pathogenesis of disseminated neoplasia in eastern Pacific Mytilus trossulus. [Ph.D. Dissertation, University of Washington]."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1038\/nature18599","article-title":"Widespread trans-mission of independent cancer lineages within multiple bivalve species","volume":"534","author":"Metzger","year":"2016","journal-title":"Nature"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Metzger, M.J., and Goff, S.P. (2016). A Sixth Modality of Infectious Disease: Contagious Cancer from Devils to Clams and Beyond. PLoS Pathog., 12.","DOI":"10.1371\/journal.ppat.1005904"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e47788","DOI":"10.7554\/eLife.47788","article-title":"A single clonal lineage of transmissible cancer identified in two marine mussel species in South America and Europe","volume":"8","author":"Giersch","year":"2019","journal-title":"eLife"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"736","DOI":"10.1111\/mec.16052","article-title":"Prevalence and polymorphism of a mussel transmissible cancer in Europe","volume":"31","author":"Hammel","year":"2021","journal-title":"Mol. Ecol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5809","DOI":"10.1038\/s41598-021-85098-5","article-title":"First description of a widespread Mytilus trossu-lus-derived bivalve transmissible cancer lineage in M. trossulus itself","volume":"11","author":"Skazina","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e66946","DOI":"10.7554\/eLife.66946","article-title":"Mitochondrial genome sequencing of marine leukaemias reveals cancer contagion between clam species in the Seas of Southern Europe","volume":"11","author":"Bruzos","year":"2022","journal-title":"eLife"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e2000222","DOI":"10.1002\/bies.202000222","article-title":"Transmissible cancers in mammals and bivalves: How many examples are there? Predictions indicate widespread occurrence","volume":"43","author":"Dujon","year":"2021","journal-title":"Bioessays"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1111\/j.1749-6632.1977.tb19287.x","article-title":"Prevalence of neoplasia in 10 New England populations of the soft-shell clam (Mya arenaria)","volume":"298","author":"Brown","year":"1977","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1111\/j.1749-6632.1977.tb19281.x","article-title":"Neoplasia in soft-shell clams (Mya arenaria) collected from oil-impacted sites","volume":"298","author":"Yevich","year":"1978","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_21","first-page":"35","article-title":"Epizootiology and distribution of transmissible sarcoma in Maryland softshell clams, Mya arenaria, 1984\u20131988","volume":"90","author":"Farley","year":"1991","journal-title":"Environ. Health Perspect."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1016\/0300-9629(92)90731-5","article-title":"Serum-cell interactions in transmission of sarcoma in the soft shell clam, Mya arenaria L","volume":"102","author":"Sunila","year":"1992","journal-title":"Comp. Biochem. Physiol. Part A: Physiol."},{"key":"ref_23","first-page":"227","article-title":"Transplantables Rundzellensarkom des Hundes","volume":"4","author":"Sticker","year":"1906","journal-title":"J. Cancer Res. Clin. Oncol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1002\/path.1700120221","article-title":"An experimental investigation of infective sarcoma of the dog, with a consideration of its relationship to cancer","volume":"12","author":"Wade","year":"1908","journal-title":"J. Pathol. Bacteriol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1111\/j.1365-2656.2012.02025.x","article-title":"Biting injuries and transmission of Tasmanian devil facial tumour disease","volume":"82","author":"Hamede","year":"2013","journal-title":"J. Anim. Ecol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"127","DOI":"10.3354\/meps12991","article-title":"Developing an eDNA toolkit to quantify broadcast spawning events of the sea scallop Placopecten magellanicus: Moving beyond fertilization assays","volume":"621","author":"Bayer","year":"2019","journal-title":"Mar. Ecol. Prog. Ser."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41467-019-14105-1","article-title":"Environmental DNA reveals seasonal shifts and potential interactions in a marine community","volume":"11","author":"Djurhuus","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"180537","DOI":"10.1098\/rsos.180537","article-title":"Water, water everywhere: Environmental DNA can unlock population structure in elusive marine species","volume":"5","author":"Parsons","year":"2018","journal-title":"R. Soc. Open Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1038\/s42003-021-02085-2","article-title":"Environmental DNA monitoring of oncogenic viral shedding and genomic profiling of sea turtle fibropapillo-matosis reveals unusual viral dynamics","volume":"4","author":"Farrell","year":"2021","journal-title":"Commun Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"20202010","DOI":"10.1098\/rspb.2020.2010","article-title":"Environmental DNA from multiple pathogens is elevated near active Atlantic salmon farms","volume":"287","author":"Shea","year":"2020","journal-title":"Proc. R. Soc. B Boil. Sci."},{"key":"ref_31","unstructured":"Maine State Legislature, Maine Office of Policy and Legal Analysis, Bentley, C., and Schneider, D. (2022, February 02). Report of the Commission to Study the Effects of Coastal and Ocean Acidification and its Existing and Potential Effects on Species that are Commercially Harvested and Grown Along the Maine Coast. Office of Policy and Legal Analysis, Available online: https:\/\/digitalmaine.com\/opla_docs\/145."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1006\/jipa.1996.4641","article-title":"Experimental field studies with Mya arenaria (Bivalvia) on the induction and effect of hematopoietic neoplasia","volume":"69","author":"Weinberg","year":"1997","journal-title":"J. Invertebr. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1007\/BF01344301","article-title":"Hematopoietic neoplasia in Mya arenaria: Prevalence and indices of physiological condition","volume":"105","author":"Leavitt","year":"1990","journal-title":"Mar. Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"24110","DOI":"10.1038\/s41598-021-03598-w","article-title":"Traits of a mussel transmissible cancer are reminiscent of a parasitic life style","volume":"11","author":"Burioli","year":"2021","journal-title":"Sci Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-020-76677-z","article-title":"Hemocytes released in seawater act as Trojan horses for spreading of bacterial infections in mussels","volume":"10","author":"Caza","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1002\/lom3.10325","article-title":"Correcting in situ chlorophyll fluorescence time-series observations for nonphotochemical quenching and tidal variability reveals nonconservative phytoplankton variability in coastal waters","volume":"17","author":"Carberry","year":"2019","journal-title":"Limnol. Oceanogr. Methods"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Wickham, H. (2016). ggplot2: Elegant Graphics for Data Analysis, Springer.","DOI":"10.1007\/978-3-319-24277-4_9"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1111\/1755-0998.12281","article-title":"The room temperature preservation of filtered Environmental DNA samples and assimilation into a phenol-chloroform-isoamyl alcohol DNA extraction","volume":"15","author":"Renshaw","year":"2015","journal-title":"Mol. Ecol. Resour."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"14175","DOI":"10.1073\/pnas.1409945111","article-title":"Activation of transcription and retrotransposition of a novel retroelement, Steamer, in neoplastic hemocytes of the mollusk Mya arenaria","volume":"111","author":"Arriagada","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/11\/3\/283\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:26:27Z","timestamp":1760135187000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/11\/3\/283"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,23]]},"references-count":39,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["pathogens11030283"],"URL":"https:\/\/doi.org\/10.3390\/pathogens11030283","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2021.12.25.473725","asserted-by":"object"}]},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,23]]}}}