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dc.contributor.authorKim, Minjien_US
dc.contributor.authorShapiro, Karenen_US
dc.contributor.authorRajal, Verónica Beatrizen_US
dc.contributor.authorPackham, Andreaen_US
dc.contributor.authorAguilar, Beatrizen_US
dc.contributor.authorRueda, Lezlieen_US
dc.contributor.authorWuertz, Stefanen_US
dc.identifier.citationKim, M., Shapiro, K., Rajal, V. B., Packham, A., Aguilar, B., Rueda, L. & Wuertz, S. (2021). Quantification of viable protozoan parasites on leafy greens using molecular methods. Food Microbiology, 99, 103816-.
dc.description.abstractProtozoan contamination in produce is of growing importance due to their capacity to cause illnesses in consumers of fresh leafy greens. Viability assays are essential to accurately estimate health risk caused by viable parasites that contaminate food. We evaluated the efficacy of reverse transcription quantitative PCR (RT-qPCR), propidium monoazide coupled with (q)PCR, and viability staining using propidium iodide through systematic laboratory spiking experiments for selective detection of viable Cryptosporidium parvum, Giardia enterica, and Toxoplasma gondii. In the presence of only viable protozoa, the RT-qPCR assays could accurately detect two to nine (oo)cysts/g spinach (in 10 g processed). When different proportions of viable and inactivated parasite were spiked, mRNA concentrations correlated with increasing proportions of viable (oo)cysts, although low levels of false-positive mRNA signals were detectable in the presence of high amounts of inactivated protozoa. Our study demonstrated that among the methods tested, RT-qPCR performed more effectively to discriminate viable from inactivated C. parvum, G. enterica and T. gondii on spinach. This application of viability methods on leafy greens can be adopted by the produce industry and regulatory agencies charged with protection of human public health to screen leafy greens for the presence of viable protozoan pathogen contamination.en_US
dc.relation.ispartofFood Microbiologyen_US
dc.rights© 2021 Elsevier Ltd. All rights reserved. This paper was published in Food Microbiology and is made available with permission of Elsevier Ltd.en_US
dc.subjectEngineering::Civil engineeringen_US
dc.titleQuantification of viable protozoan parasites on leafy greens using molecular methodsen_US
dc.typeJournal Articleen
dc.contributor.schoolSchool of Civil and Environmental Engineeringen_US
dc.contributor.researchSingapore Centre for Environmental Life Sciences and Engineering (SCELSE)en_US
dc.description.versionSubmitted/Accepted versionen_US
dc.description.acknowledgementThis work was supported by the Center for Produce Safety (California Department of Food and Agriculture grant# SBC15064CPS).en_US
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