{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T04:36:16Z","timestamp":1775536576749,"version":"3.50.1"},"reference-count":40,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2021,5,28]],"date-time":"2021-05-28T00:00:00Z","timestamp":1622160000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJERPH"],"abstract":"<jats:p>Detecting environmental exposures and mitigating their impacts are growing global public health challenges. Antibody tests show great promise and have emerged as fundamental tools for large-scale exposure studies. Here, we apply, demonstrate and validate the utility of a salivary antibody multiplex immunoassay in measuring antibody prevalence and immunoconversions to six pathogens commonly found in the environment. The study aimed to assess waterborne infections in consenting beachgoers recreating at an Iowa riverine beach by measuring immunoglobulin G (IgG) antibodies against select pathogens in serially collected saliva samples. Results showed that nearly 80% of beachgoers had prior exposures to at least one of the targeted pathogens at the beginning of the study. Most of these exposures were to norovirus GI.1 (59.41%), norovirus GII.4 (58.79%) and Toxoplasma gondii (22.80%) and over half (56.28%) of beachgoers had evidence of previous exposure to multiple pathogens. Of individuals who returned samples for each collection period, 6.11% immunoconverted to one or more pathogens, largely to noroviruses (GI.1: 3.82% and GII.4: 2.29%) and T. gondii (1.53%). Outcomes of this effort illustrate that the multiplex immunoassay presented here serves as an effective tool for evaluating health risks by providing valuable information on the occurrence of known and emerging pathogens in population surveillance studies.<\/jats:p>","DOI":"10.3390\/ijerph18115797","type":"journal-article","created":{"date-parts":[[2021,5,28]],"date-time":"2021-05-28T11:33:20Z","timestamp":1622201600000},"page":"5797","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Salivary Antibodies against Multiple Environmental Pathogens Found in Individuals Recreating at an Iowa Beach"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4948-884X","authenticated-orcid":false,"given":"Swinburne A. J.","family":"Augustine","sequence":"first","affiliation":[{"name":"Center for Public Health and Environmental Assessment, United States Environmental Protection Agency, Cincinnati, OH 45268, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9881-3293","authenticated-orcid":false,"given":"Tarsha N.","family":"Eason","sequence":"additional","affiliation":[{"name":"Center for Environmental Methods and Measurement, United States Environmental Protection Agency, Athens, GA 30605, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tim","family":"Wade","sequence":"additional","affiliation":[{"name":"Center for Public Health and Environmental Assessment, United States Environmental Protection Agency, Research Triangle Park, NC 27709, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shannon M.","family":"Griffin","sequence":"additional","affiliation":[{"name":"Center for Public Health and Environmental Assessment, United States Environmental Protection Agency, Cincinnati, OH 45268, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elizabeth","family":"Sams","sequence":"additional","affiliation":[{"name":"Center for Public Health and Environmental Assessment, United States Environmental Protection Agency, Research Triangle Park, NC 27709, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kaneatra","family":"Simmons","sequence":"additional","affiliation":[{"name":"Department of Arts and Sciences\/Learning Support, Fort Valley State University, Fort Valley, GA 31030, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Malini","family":"Ramudit","sequence":"additional","affiliation":[{"name":"Oak Ridge Institute for Science Education, Oak Ridge, TN 37831, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin","family":"Oshima","sequence":"additional","affiliation":[{"name":"Center for Environmental Methods and Measurement, United States Environmental Protection Agency, Cincinnati, OH 45268, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alfred","family":"Dufour","sequence":"additional","affiliation":[{"name":"Center for Environmental Methods and Measurement, United States Environmental Protection Agency, Cincinnati, OH 45268, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1007\/s00248-015-0629-0","article-title":"A Perspective on the Global Pandemic of Waterborne Disease","volume":"76","author":"Ford","year":"2018","journal-title":"Microb. Ecol."},{"key":"ref_2","unstructured":"Wade, T.J., Krueger, W., Sams, E., Converse, R., Hudgens, E., and Dufour, A. (2015, January 16\u201319). Health Risks Associated with Swimming at an Inland River. Proceedings of the Society for Epidemiologic Research Conference, Denver, CO, USA."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1007\/s40572-016-0096-x","article-title":"Use of Pathogen-Specific Antibody Biomarkers to Estimate Waterborne Infections in Population-Based Settings","volume":"3","author":"Exum","year":"2016","journal-title":"Curr. Environ. Health Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1016\/j.cmi.2016.07.016","article-title":"Speculations on the Clinical Significance of Asymptomatic Viral Infections","volume":"22","author":"Gentile","year":"2016","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"362","DOI":"10.7326\/M20-3012","article-title":"Prevalence of Asymptomatic Sars-Cov-2 Infection: A Narrative Review","volume":"173","author":"Oran","year":"2020","journal-title":"Ann. Intern. Med."},{"key":"ref_6","first-page":"e54415","article-title":"Developing a Salivary Antibody Multiplex Immunoassay to Measure Human Exposure to Environmental Pathogens","volume":"115","author":"Augustine","year":"2016","journal-title":"J. Vis. Exp."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1196\/annals.1384.012","article-title":"Do Salivary Antibodies Reliably Reflect Both Mucosal and Systemic Immunity?","volume":"1098","author":"Brandtzaeg","year":"2007","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.jim.2010.11.005","article-title":"Development of a Multiplex Microsphere Immunoassay for the Quantitation of Salivary Antibody Responses to Selected Waterborne Pathogens","volume":"364","author":"Griffin","year":"2011","journal-title":"J. Immunol. Methods"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1080\/1354750X.2016.1265009","article-title":"The Utility of Saliva for the Assessment of Anti-Pneumococcal Antibodies: Investigation of Saliva as a Marker of Antibody Status in Serum","volume":"23","author":"Heaney","year":"2018","journal-title":"Biomarkers"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1373\/clinchem.2010.152124","article-title":"Oral Fluid Testing: Promises and Pitfalls","volume":"57","author":"Huestis","year":"2011","journal-title":"Clin. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Rodenburg, G.D., Sanders, E.A.M., Van Gils, E.J.M., Veenhoven, R.H., Zborowski, T., Dobbelsteen, G.P.J.M.V.D., Bloem, A.C., Berbers, G.A.M., and Bogaert, D. (2012). Salivary Immune Responses to the 7-Valent Pneumococcal Conjugate Vaccine in the First 2 Years of Life. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0046916"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1234","DOI":"10.1093\/infdis\/jiy638","article-title":"Minimally Invasive Saliva Testing to Monitor Norovirus Infection in Community Settings","volume":"219","author":"Pisanic","year":"2018","journal-title":"J. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"84","DOI":"10.3389\/fpubh.2017.00084","article-title":"Immunoprevalence to Six Waterborne Pathogens in Beachgoers at Boqueron Beach, Puerto Rico: Application of a Microsphere-Based Salivary Antibody Multiplex Immunoassay","volume":"5","author":"Augustine","year":"2017","journal-title":"Front. Public Health"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.jim.2015.05.001","article-title":"Application of Salivary Antibody Immunoassays for the Detection of Incident Infections with Norwalk Virus in a Group of Volunteers","volume":"424","author":"Griffin","year":"2015","journal-title":"J. Immunol. Methods"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"231","DOI":"10.3389\/fpubh.2019.00231","article-title":"Visitors to a Tropical Marine Beach Show Evidence of Immunoconversions to Multiple Waterborne Pathogens","volume":"7","author":"Simmons","year":"2019","journal-title":"Front. Public Health"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Wade, T.J., Augustine, S.A.J., Griffin, S.M., Sams, E.A., Oshima, K.H., Egorov, A.I., Simmons, K.J., Eason, T.N., and Dufour, A.P. (2018). Asymptomatic Norovirus Infection Associated with Swimming at a Tropical Beach: A Prospective Cohort Study. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0195056"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-019-56040-7","article-title":"Application of a Multiplex Salivary Immunoassay to Detect Sporadic Incident Norovirus Infections","volume":"9","author":"Wade","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Augustine, S.A.J., Eason, T.N., Simmons, K.J., Griffin, S.M., Curioso, C.L., Ramudit, M.K.D., Sams, E.A., Oshima, K.H., Dufour, A., and Wade, T.J. (2020). Rapid Salivary Igg Antibody Screening for Hepatitis A. J. Clin. Microbiol., 58.","DOI":"10.1128\/JCM.00358-20"},{"key":"ref_19","unstructured":"Anonymous (2020, August 26). The Power of Multiplexing. Thermofisher, Available online: https:\/\/www.thermofisher.com\/content\/dam\/LifeTech\/global\/technical-reference-library\/s2s\/dbourdon\/The%20power%20of%20multiplexing.pdf."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jim.2015.06.002","article-title":"Statistical Approaches to Developing a Multiplex Immunoassay for Determining Human Exposure to Environmental Pathogens","volume":"425","author":"Augustine","year":"2015","journal-title":"J. Immunol. Methods"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.tibtech.2015.03.002","article-title":"Digital detection of biomarkers assisted by nanoparticles: Application to diagnostics","volume":"33","author":"Cretich","year":"2015","journal-title":"Trends Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1111\/jtm.12092","article-title":"Seroepidemiology of norovirus-associated travelers\u2019 diarrhea","volume":"21","author":"Ajami","year":"2014","journal-title":"J. Travel Med."},{"key":"ref_23","first-page":"1028","article-title":"Diagnosis of Norwalk Virus Infection by Indirect Enzyme Immunoassay Detection of Salivary Antibodies to Recombinant Norwalk Virus Antigen","volume":"11","author":"Moe","year":"2004","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2866","DOI":"10.1128\/jcm.31.11.2866-2872.1993","article-title":"Detection of antibody to recombinant Norwalk virus antigen in specimens from outbreaks of gastroenteritis","volume":"31","author":"Monroe","year":"1993","journal-title":"J. Clin. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1093\/aje\/154.4.357","article-title":"Toxoplasma gondii Infection in the United States: Seroprevalence and Risk Factors","volume":"154","author":"Jones","year":"2001","journal-title":"Am. J. Epidemiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1177\/003335491112600408","article-title":"Seroprevalence of Hepatitis A Virus Antibodies in the U.S.: Results from the National Health and Nutrition Examination Survey","volume":"126","author":"Klevens","year":"2011","journal-title":"Public Health Rep."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Krueger, W.S., Hilborn, E.D., Converse, R.R., and Wade, T.J. (2014). Drinking water source and human Toxoplasma gondii infection in the United States: A cross-sectional analysis of NHANES data. BMC Public Health, 14.","DOI":"10.1186\/1471-2458-14-711"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3","DOI":"10.5812\/jjm.20783","article-title":"Determining the Role of Helicobacter pylori in Chronic Sinus Infections Using the Polymerase Chain Reaction","volume":"8","author":"Nikakhlagh","year":"2015","journal-title":"Jundishapur J. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2278","DOI":"10.1128\/JCM.00381-10","article-title":"Detection of Campylobacter jejuni by Culture and Real-Time PCR in a French Cohort of Patients with Guillain-Barre Syndrome","volume":"48","author":"Orlikowski","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e12453","DOI":"10.1111\/hel.12453","article-title":"Seroprevalence of Helicobacter pylori Hispanics living in Puerto Rico: A population-based study","volume":"23","author":"Sevilla","year":"2018","journal-title":"Helicobacter"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2847","DOI":"10.1007\/s00216-011-5047-7","article-title":"Lanthanide-based time-resolved luminescence immunoassays","volume":"400","author":"Hagan","year":"2011","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.1111\/j.1432-2277.2009.00927.x","article-title":"The aging of the immune system","volume":"22","author":"Weiskopf","year":"2009","journal-title":"Transpl. Int."},{"key":"ref_33","first-page":"57","article-title":"B-cell memory in malaria: Myths and realities","volume":"293","author":"Ndungu","year":"2019","journal-title":"Immunol. Rev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1001\/jama.1983.03330250028023","article-title":"Effect of Immunoglobulin on Hepatitis A in Day-care Centers","volume":"249","author":"Hadler","year":"1983","journal-title":"JAMA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"102255","DOI":"10.1016\/j.isci.2021.102255","article-title":"Seasonal and daytime variation in multiple immune parameters in humans: Evidence from 329,261 participants of the UK Biobank cohort","volume":"24","author":"Wyse","year":"2021","journal-title":"Iscience"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"786","DOI":"10.1093\/aje\/kwh107","article-title":"A population-based seroprevalence study of hepatitis A virus using oral fluid in England and Wales","volume":"159","author":"Edmunds","year":"2004","journal-title":"Am. J. Epidemiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1017\/S0950268803001493","article-title":"A novel method of oral fluid collection to monitor immunity to common viral infections","volume":"132","author":"Edmunds","year":"2004","journal-title":"Epidemiol. Infect."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1128\/CDLI.6.4.577-580.1999","article-title":"A Simple Saliva-Based Test for Detecting Antibodies to Human Immunodeficiency Virus","volume":"6","author":"Schramm","year":"1999","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"ref_39","first-page":"161","article-title":"Salivary antibody testing in a school outbreak of hepatitis A","volume":"109","author":"Stuart","year":"1992","journal-title":"Epidemiol. Infect."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Yap, G., Sil, B.K., and Ng, L.-C. (2011). Use of Saliva for Early Dengue Diagnosis. PLoS Negl. Trop. Dis., 5.","DOI":"10.1371\/journal.pntd.0001046"}],"container-title":["International Journal of Environmental Research and Public Health"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-4601\/18\/11\/5797\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:09:46Z","timestamp":1760162986000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-4601\/18\/11\/5797"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,28]]},"references-count":40,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2021,6]]}},"alternative-id":["ijerph18115797"],"URL":"https:\/\/doi.org\/10.3390\/ijerph18115797","relation":{},"ISSN":["1660-4601"],"issn-type":[{"value":"1660-4601","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,28]]}}}