title: Development of a combined in vitro cell culture--quantitative PCR assay for evaluating the disinfection performance of pulsed light for treating the waterborne enteroparasite Giardia lamblia.
abstract: Giardia lamblia is a flagellated protozoan parasite that is recognised as a frequent cause of water-borne disease in humans and animals. We report for the first time on the use of a combined in vitro HCT-8 cell culture-quantitative PCR assay for evaluating the efficacy of using pulsed UV light for treating G. lamblia parasites. Findings showed that current methods that are limited to using vital stains before and after cyst excystation are not appropriate for monitoring or evaluating cyst destruction post PUV-treatments. Use of the human ileocecal HCT-8 cell line was superior to that of the human colon Caco-2 cell line for in vitro culture and determining PUV sensitivity of treated cysts. G. lamblia cysts were also shown to be more resistant to PUV irradiation compared to treating similar numbers of Cryptosporidium parvum oocysts. These observations also show that the use of this HCT-8 cell culture assay may replace use of animal models for determining disinfection performances of PUV for treating both C. parvum and G. lamblia.
authors.affiliation: Bioscience Research Institute, Athlone Institute of Technology, Dublin Rd, Athlone, Westmeath, Ireland. Electronic address: mgarvey@ait.ie.
authors.forename: Mary
authors.initials: M
authors.lastname: Garvey
authors.affiliation: Bioscience Research Institute, Athlone Institute of Technology, Dublin Rd, Athlone, Westmeath, Ireland.
authors.forename: Alessia
authors.initials: A
authors.lastname: Stocca
authors.affiliation: Bioscience Research Institute, Athlone Institute of Technology, Dublin Rd, Athlone, Westmeath, Ireland.
authors.forename: Neil
authors.initials: N
authors.lastname: Rowan
mesh_terms: D018938:Caco-2 Cells; D045744:Cell Line, Tumor; D016785:Cryptosporidium parvum / Q000235:genetics / Q000302:isolation & purification / Q000528:radiation effects; D016054:DNA, Protozoan / Q000032:analysis; D004203:Disinfection / Q000379:methods* / Q000592:standards; D016829:Giardia lamblia / Q000235:genetics / Q000302:isolation & purification / Q000528:radiation effects*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013997:Time Factors; D014466:Ultraviolet Rays; D014867:Water / Q000469:parasitology*
chemical_list: D016054:DNA, Protozoan; D014867:Water
keywords: Cell culture; Giardia lamblia; In vitro; Pulsed UV inactivation; Real time PCR