{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:52:00Z","timestamp":1787028720068,"version":"build-2736575974"},"reference-count":49,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T00:00:00Z","timestamp":1737590400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Research Foundation of South Africa","award":["115566"],"award-info":[{"award-number":["115566"]}]},{"name":"University of the Free State","award":["115566"],"award-info":[{"award-number":["115566"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Wastewater surveillance is a promising tool to monitor potential outbreaks and determine the disease burden within a community. This system has been extensively used to monitor polio and COVID-19 infection levels, yet few attempts have been made to apply it to monitoring pathogenic yeast. This study aimed to investigate the application of wastewater surveillance for potentially pathogenic yeast in wastewater treatment plant influent. This was done by comparing culture-dependent data with culture-independent data and investigating the fluconazole concentration in wastewater. Additional studies on the growth of isolated strains were conducted. We found that a multiplex PCR system to detect multiple yeasts holds promise as a molecular detection tool for wastewater surveillance. Culture-dependent results indicated that Candida spp. specifically C. krusei and C. glabrata, were most prominent. Growth studies supported that these species grow well in this environment while the less frequently isolated yeasts grew poorly. The data from culture-dependent and independent techniques showed some correlation, with similar species being identified with both, further promoting the use of molecular tools for surveillance. This study highlights the presence of potentially pathogenic yeasts in wastewater, which may indicate the prevalence of these yeasts in the environment or community. This wastewater may also be a potential source of infection for persons encountering it due to poor wastewater management.<\/jats:p>","DOI":"10.3390\/jof11020086","type":"journal-article","created":{"date-parts":[[2025,1,23]],"date-time":"2025-01-23T09:01:13Z","timestamp":1737622873000},"page":"86","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Culture-Dependent and -Independent Wastewater Surveillance for Multiple Pathogenic Yeasts"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2212-543X","authenticated-orcid":false,"given":"Tyla","family":"Baker","sequence":"first","affiliation":[{"name":"Department of Microbiology and Biochemistry, University of the Free State, P.O. Box 339, Bloemfontein 9301, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5795-5313","authenticated-orcid":false,"given":"Phillip Armand","family":"Bester","sequence":"additional","affiliation":[{"name":"School of Pathology, University of the Free State, P.O. Box 339, Bloemfontein 9301, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0182-1084","authenticated-orcid":false,"given":"Olihile Moses","family":"Sebolai","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Biochemistry, University of the Free State, P.O. Box 339, Bloemfontein 9301, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacobus","family":"Albertyn","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Biochemistry, University of the Free State, P.O. Box 339, Bloemfontein 9301, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6928-5663","authenticated-orcid":false,"given":"Carolina Henritta","family":"Pohl","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Biochemistry, University of the Free State, P.O. Box 339, Bloemfontein 9301, South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e428","DOI":"10.1016\/S1473-3099(23)00692-8","article-title":"Global incidence and mortality of severe fungal disease","volume":"24","author":"Denning","year":"2024","journal-title":"Lancet Infect. Dis."},{"key":"ref_2","unstructured":"World Health Organization (2022). WHO Fungal Priority Pathogens List to Guide Research, Development and Public Health Action, World Health Organization."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1038\/s41579-019-0322-2","article-title":"Drug resistance and tolerance in fungi","volume":"18","author":"Berman","year":"2020","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Sanglard, D. (2016). Emerging threats in antifungal-resistant fungal pathogens. Front. Med., 3.","DOI":"10.3389\/fmed.2016.00011"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1299","DOI":"10.1111\/1462-2920.14055","article-title":"Emergence of azole resistant Aspergillus fumigatus and one health: Time to implement environmental stewardship","volume":"20","author":"Chowdhary","year":"2018","journal-title":"Environ. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1016\/S1473-3099(09)70265-8","article-title":"Azole resistance in Aspergillus fumigatus: A side-effect of environmental fungicide use?","volume":"9","author":"Verweij","year":"2009","journal-title":"Lancet Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1016\/j.jenvman.2010.11.011","article-title":"Removal of heavy metal ions from wastewater: A review","volume":"92","author":"Fu","year":"2011","journal-title":"J. Environ. Manag."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2932","DOI":"10.5897\/AJB2007.000-2462","article-title":"Wastewater treatment plants as a source of microbial pathogens in receiving watersheds","volume":"6","author":"Okoh","year":"2007","journal-title":"Afr. J. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/S0011-9164(04)00169-9","article-title":"Removal of heavy metals from wastewater by membrane processes: A comparative study","volume":"154","author":"Qdais","year":"2004","journal-title":"Desalination"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Yang, J., Hou, B., Wang, J., Tian, B., Bi, J., Wang, N., Li, X., and Huang, X. (2019). Nanomaterials for the removal of heavy metals from wastewater. Nanomaterials, 9.","DOI":"10.3390\/nano9030424"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e00591-22","DOI":"10.1128\/mbio.00591-22","article-title":"Microbiome analysis for wastewater surveillance during COVID-19","volume":"13","author":"Brumfield","year":"2022","journal-title":"mBio"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"116160","DOI":"10.1016\/j.watres.2020.116160","article-title":"Identification of multiple potential viral diseases in a large urban center using wastewater surveillance","volume":"184","author":"McCall","year":"2020","journal-title":"Water Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"20586","DOI":"10.2807\/1560-7917.ES2013.18.38.20586","article-title":"Insidious reintroduction of wild poliovirus into Israel","volume":"18","author":"Anis","year":"2013","journal-title":"Eurosurveillance"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1126\/science.90.2333.258","article-title":"Poliomyelitic virus in sewage","volume":"90","author":"Paul","year":"1939","journal-title":"Science"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"o1546","DOI":"10.1136\/bmj.o1546","article-title":"Poliovirus is detected in sewage from north and east London","volume":"377","author":"Wise","year":"2022","journal-title":"BMJ"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"138875","DOI":"10.1016\/j.scitotenv.2020.138875","article-title":"Computational analysis of SARS-CoV-2\/COVID-19 surveillance by wastewater-based epidemiology locally and globally: Feasibility, economy, opportunities and challenges","volume":"730","author":"Hart","year":"2020","journal-title":"Sci. Total Environ."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1164","DOI":"10.1038\/s41587-020-0684-z","article-title":"Measurement of SARS-CoV-2 RNA in wastewater tracks community infection dynamics","volume":"38","author":"Peccia","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"117400","DOI":"10.1016\/j.watres.2021.117400","article-title":"Wastewater surveillance of SARS-CoV-2 across 40 U.S. states from February to June 2020","volume":"202","author":"Wu","year":"2021","journal-title":"Water Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"165459","DOI":"10.1016\/j.scitotenv.2023.165459","article-title":"Detection of the clinically persistent, pathogenic yeast spp. Candida auris from hospital and municipal wastewater in Miami-Dade County, Florida","volume":"898","author":"Babler","year":"2023","journal-title":"Sci. Total Environ."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1755","DOI":"10.1021\/acs.est.2c07763","article-title":"Community-scale wastewater surveillance of Candida auris during an ongoing outbreak in southern Nevada","volume":"57","author":"Barber","year":"2023","journal-title":"Environ. Sci. Technol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"422","DOI":"10.3201\/eid2902.221523","article-title":"Candida auris discovery through community wastewater surveillance during healthcare outbreak, Nevada, USA, 2022","volume":"29","author":"Rossi","year":"2023","journal-title":"Emerg. Infect. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zulli, A., Chan, E.M.G., Shelden, B., Duong, D., Xu, X.S., White, B.J., Wolfe, M.K., and Boehm, A.B. (2024). Prospective study of Candida auris nucleic acids in wastewater solids in 190 wastewater treatment plants in the United States suggests widespread occurrence. mBio, 15.","DOI":"10.1128\/mbio.00908-24"},{"key":"ref_23","unstructured":"(2024, September 29). COVID-19 Prevention Research Programme: Wastewater Surveillance for SARS-CoV-2. Wastewater Sampling Guide. Available online: https:\/\/www.samrc.ac.za\/wbe\/SARS-CoV-2WastewaterCollectionManual.pdf."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Harrigan, W.F., and McCance, E.M. (1966). Laboratory Methods in Microbiology, Academic Press.","DOI":"10.1016\/B978-1-4832-3205-8.50005-2"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"7371","DOI":"10.1007\/s00253-017-8471-6","article-title":"Comparison of EMA-, PMA- and DNase qPCR for the determination of microbial cell viability","volume":"101","author":"Reyneke","year":"2017","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.diagmicrobio.2018.09.007","article-title":"YEAST PANEL multiplex PCR for identification of clinically important yeast species: Stepwise diagnostic strategy, useful for developing countries","volume":"93","author":"Arastehfar","year":"2019","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.funeco.2015.10.001","article-title":"Yeasts and yeast-like fungi in tap water and groundwater, and their transmission to household appliances","volume":"20","author":"Zalar","year":"2016","journal-title":"Fungal Ecol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"129718","DOI":"10.1016\/j.chemosphere.2021.129718","article-title":"Antifungal agents, yeast abundance and diversity in surface water: Potential risks to water users","volume":"274","author":"Monapathi","year":"2021","journal-title":"Chemosphere"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1582","DOI":"10.1590\/S1517-83822012000400043","article-title":"Water quality and diversity of yeast from tropical lakes and rivers from the Rio Doce basin in Southeastern Brazil","volume":"43","author":"Medeiros","year":"2012","journal-title":"Braz. J. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3813","DOI":"10.1016\/j.watres.2009.05.024","article-title":"Occurrence of filamentous fungi and yeasts in three different drinking water sources","volume":"43","author":"Pereira","year":"2009","journal-title":"Water Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1590\/S1516-89132007000100001","article-title":"D Yeasts and filamentous fungi in bottled mineral water and tap water from municipal supplies","volume":"50","author":"Yamaguchi","year":"2007","journal-title":"Braz. Arch. Biol. Technol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Ayanbimpe, G.M., Abbah, V.E., and Ior, C.A. (2012). Yeasts and yeast-like fungal contaminants of water used for domestic purposes in Jos, Nigeria. Microbiol. Res., 3.","DOI":"10.4081\/mr.2012.e24"},{"key":"ref_33","first-page":"129","article-title":"Fungi associated with the activated-sludge process of sewage treatment at the Lebanon, Ohio, sewage treatment plant","volume":"70","author":"Cooke","year":"1970","journal-title":"Ohio J. Sci."},{"key":"ref_34","first-page":"249","article-title":"The occurrence of fungi in activated sludge","volume":"40","author":"Cooke","year":"1969","journal-title":"Mycopathologia"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1016\/j.desal.2005.03.085","article-title":"The comparative mycological analysis of wastewater and sewage sludges from selected wastewater treatment plants","volume":"185","author":"Kacprzak","year":"2005","journal-title":"Desalination"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1016\/j.biortech.2012.11.101","article-title":"Extracellular enzyme production and phylogenetic distribution of yeasts in wastewater treatment systems","volume":"129","author":"Yang","year":"2013","journal-title":"Bioresour. Technol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Zein, S.E.I., Hindy, J.R., and Kanj, S.S. (2020). Invasive Saprochaete infections: An emerging threat to immunocompromised patients. Pathogens, 9.","DOI":"10.3390\/pathogens9110922"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"493","DOI":"10.2323\/jgam.34.493","article-title":"Candida palmioleophila sp. nov., a yeast capable of assimilating crude palm oil, formerly identified as Torulopsis candida","volume":"34","author":"Nakase","year":"1988","journal-title":"J. Gen. Appl. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1128\/JCM.02071-10","article-title":"Candida palmioleophila: Characterization of a previously overlooked pathogen and its unique susceptibility profile in comparison with five related species","volume":"49","author":"Jensen","year":"2011","journal-title":"J. Clin. Microbiol."},{"key":"ref_40","first-page":"191","article-title":"Candida palmioleophila Carba14 is capable of degrading carbamazepine","volume":"28","year":"2023","journal-title":"Bioremediation J."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Parslow, B.Y., and Thornton, C.R. (2022). Continuing shifts in epidemiology and antifungal susceptibility highlight the need for improved disease management of invasive candidiasis. Microorganisms, 10.","DOI":"10.3390\/microorganisms10061208"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/j.bjm.2017.09.002","article-title":"The potential of the newly isolated thermotolerant yeast Pichia kudriavzevii RZ8-1 for high-temperature ethanol production","volume":"49","author":"Chamnipa","year":"2018","journal-title":"Braz. J. Microbiol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Chu, Y., Li, M., Jin, J., Dong, X., Xu, K., Jin, L., Qiao, Y., and Ji, H. (2023). Advances in the application of the non-conventional yeast Pichia kudriavzevii in food and biotechnology industries. J. Fungi, 9.","DOI":"10.3390\/jof9020170"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Douglass, A.P., Offei, B., Braun-Galleani, S., Coughlan, A.Y., Martos, A.A.R., Ortiz-Merino, R.A., Byrne, K.P., and Wolfe, K.H. (2018). Population genomics shows no distinction between pathogenic Candida krusei and environmental Pichia kudriavzevii: One species, four names. PloS Pathog., 14.","DOI":"10.1371\/journal.ppat.1007138"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1016\/j.ijantimicag.2017.05.012","article-title":"Molecular basis of antifungal drug resistance in yeasts","volume":"50","author":"Morio","year":"2017","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Aboutalebian, S., Ahmadikia, K., Fakhim, H., Chabavizadeh, J., Okhovat, A., Nikaeen, M., and Mirhendi, H. (2021). Direct detection and identification of the most common bacteria and fungi causing otitis externa by a stepwise multiplex PCR. Front. Cell Infect. Microbiol., 11.","DOI":"10.3389\/fcimb.2021.644060"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Corr\u00eaa-Moreira, D., da Costa, G.L., de Lima Neto, R.G., Pinto, T., Salom\u00e3o, B., Machado Fumian, T., Ferreira Mannarino, C., Prado, T., Pereira Miagostovich, M., and de Souza Ramos, L. (2024). Screening of Candida spp. in wastewater in Brazil during COVID-19 pandemic: Workflow for monitoring fungal pathogens. BMC Biotechnol., 24.","DOI":"10.1186\/s12896-024-00868-z"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"119599","DOI":"10.1016\/j.watres.2023.119599","article-title":"Health risk posed by direct ingestion of yeasts from polluted river water","volume":"231","author":"Steffen","year":"2023","journal-title":"Water Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"144302","DOI":"10.1016\/j.jclepro.2024.144302","article-title":"Novel insights on biofilm development in sewers: Cross-kingdom exchange of quorum-sensing signaling molecules","volume":"484","author":"Ren","year":"2024","journal-title":"J. Clean. Prod."}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/11\/2\/86\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:34:16Z","timestamp":1759919656000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/11\/2\/86"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,23]]},"references-count":49,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["jof11020086"],"URL":"https:\/\/doi.org\/10.3390\/jof11020086","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,23]]}}}