Tags

Type your tag names separated by a space and hit enter

Investigations of the relationship between use of in vitro cell culture-quantitative PCR and a mouse-based bioassay for evaluating critical factors affecting the disinfection performance of pulsed UV light for treating Cryptosporidium parvum oocysts in saline.
J Microbiol Methods. 2010 Mar; 80(3):267-73.JM

Abstract

Cryptosporidium parvum is an enteric coccidian parasite that is recognised as a frequent cause of water-borne disease in humans. We report for the first time on use of the in vitro HCT-8 cell culture-quantitative PCR (qPCR) assay and the in vivo SCID-mouse bioassay for evaluating critical factors that reduce or eliminate infectivity of C. parvum after irradiating oocysts in saline solution under varying operational conditions with pulsed UV light. Infections post UV treatments were detected by immunofluorescence (IF) microscopy and by quantitative PCR in cell culture, and by IF staining of faeces and by hematoxylin and eosin staining of intestinal villi in mice. There was a good agreement between using cell culture-qPCR and the mouse assay for determining reduction or elimination of C. parvum infectivity as a consequence of varying UV operating conditions. Reduction in infectivity depended on the intensity of lamp discharge energy applied, amount of pulsing and population size of oocysts (P < or = 0.05). Conventional radiometer was unable to measure fluence or UV dose in saline samples due to the ultra-short non-continuous nature of the high-energy light pulses. Incorporation of humic acid at a concentration above that found in surface water (i.e., < or =10 ppm) did not significantly affect PUV disinfection capability irrespective of parameters tested (P < or = 0.05). These observations show that use of this HCT-8 cell culture assay is equivalent to using the 'gold standard' mouse-based infectivity assay for determining disinfection performances of PUV for treating C. parvum in saline solution.

Authors+Show Affiliations

Department of Nursing and Health Science, Athlone Institute of Technology, Ireland.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

20096310

Citation

Garvey, Mary, et al. "Investigations of the Relationship Between Use of in Vitro Cell Culture-quantitative PCR and a Mouse-based Bioassay for Evaluating Critical Factors Affecting the Disinfection Performance of Pulsed UV Light for Treating Cryptosporidium Parvum Oocysts in Saline." Journal of Microbiological Methods, vol. 80, no. 3, 2010, pp. 267-73.
Garvey M, Farrell H, Cormican M, et al. Investigations of the relationship between use of in vitro cell culture-quantitative PCR and a mouse-based bioassay for evaluating critical factors affecting the disinfection performance of pulsed UV light for treating Cryptosporidium parvum oocysts in saline. J Microbiol Methods. 2010;80(3):267-73.
Garvey, M., Farrell, H., Cormican, M., & Rowan, N. (2010). Investigations of the relationship between use of in vitro cell culture-quantitative PCR and a mouse-based bioassay for evaluating critical factors affecting the disinfection performance of pulsed UV light for treating Cryptosporidium parvum oocysts in saline. Journal of Microbiological Methods, 80(3), 267-73. https://doi.org/10.1016/j.mimet.2010.01.017
Garvey M, et al. Investigations of the Relationship Between Use of in Vitro Cell Culture-quantitative PCR and a Mouse-based Bioassay for Evaluating Critical Factors Affecting the Disinfection Performance of Pulsed UV Light for Treating Cryptosporidium Parvum Oocysts in Saline. J Microbiol Methods. 2010;80(3):267-73. PubMed PMID: 20096310.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Investigations of the relationship between use of in vitro cell culture-quantitative PCR and a mouse-based bioassay for evaluating critical factors affecting the disinfection performance of pulsed UV light for treating Cryptosporidium parvum oocysts in saline. AU - Garvey,Mary, AU - Farrell,Hugh, AU - Cormican,Martin, AU - Rowan,Neil, Y1 - 2010/01/21/ PY - 2009/12/01/received PY - 2010/01/07/revised PY - 2010/01/09/accepted PY - 2010/1/26/entrez PY - 2010/1/26/pubmed PY - 2010/5/12/medline SP - 267 EP - 73 JF - Journal of microbiological methods JO - J Microbiol Methods VL - 80 IS - 3 N2 - Cryptosporidium parvum is an enteric coccidian parasite that is recognised as a frequent cause of water-borne disease in humans. We report for the first time on use of the in vitro HCT-8 cell culture-quantitative PCR (qPCR) assay and the in vivo SCID-mouse bioassay for evaluating critical factors that reduce or eliminate infectivity of C. parvum after irradiating oocysts in saline solution under varying operational conditions with pulsed UV light. Infections post UV treatments were detected by immunofluorescence (IF) microscopy and by quantitative PCR in cell culture, and by IF staining of faeces and by hematoxylin and eosin staining of intestinal villi in mice. There was a good agreement between using cell culture-qPCR and the mouse assay for determining reduction or elimination of C. parvum infectivity as a consequence of varying UV operating conditions. Reduction in infectivity depended on the intensity of lamp discharge energy applied, amount of pulsing and population size of oocysts (P < or = 0.05). Conventional radiometer was unable to measure fluence or UV dose in saline samples due to the ultra-short non-continuous nature of the high-energy light pulses. Incorporation of humic acid at a concentration above that found in surface water (i.e., < or =10 ppm) did not significantly affect PUV disinfection capability irrespective of parameters tested (P < or = 0.05). These observations show that use of this HCT-8 cell culture assay is equivalent to using the 'gold standard' mouse-based infectivity assay for determining disinfection performances of PUV for treating C. parvum in saline solution. SN - 1872-8359 UR - https://www.unboundmedicine.com/medline/citation/20096310/Investigations_of_the_relationship_between_use_of_in_vitro_cell_culture_quantitative_PCR_and_a_mouse_based_bioassay_for_evaluating_critical_factors_affecting_the_disinfection_performance_of_pulsed_UV_light_for_treating_Cryptosporidium_parvum_oocysts_in_saline_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0167-7012(10)00019-9 DB - PRIME DP - Unbound Medicine ER -