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Journal of Pathogens
Volume 2014, Article ID 408204, 10 pages
http://dx.doi.org/10.1155/2014/408204
Research Article

Effective Concentration and Detection of Cryptosporidium, Giardia, and the Microsporidia from Environmental Matrices

1Center for Environmental Diagnostics and Bioremediation, University of West Florida, 11,000 University Parkway, Building 58, Room 70, Pensacola, FL 32514, USA
2Water Quality Laboratory, City of Tampa Water Department, 7125 North 30th Street, Tampa, FL 33610, USA

Received 31 May 2014; Revised 11 August 2014; Accepted 21 August 2014; Published 10 September 2014

Academic Editor: Nongnuch Vanittanakom

Copyright © 2014 Joseph A. Moss et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Cryptosporidium spp., Giardia spp., and members of Microsporidia are enteropathogenic parasites of humans and animals, producing asymptomatic to severe intestinal infections. To circumvent various impediments associated with current detection methods, we tested a method providing multistage purification and separation in a single, confined step. Standard real-time PCR was used as a detection method. Samples spiked with C. parvum and G. intestinalis were split for comparison to standard Method 1623. Results were equivalent to immunomagnetic procedures for Cryptosporidium, and Giardia. Overall percent recovery for Cryptosporidium with Method 1623 averaged 26.89% (std 21.44%; min = 0%; max = 73%) and was similar but less variable for qPCR method at an estimated average of 27.67 (std 17.65%; min = 5%; max = 63%). For Giardia, Method 1623 had an overall average recovery of 27.11% (std 17.98%; min = 1%; max = 58%), while multistage purification and qPCR had an estimated lower overall recovery at 18.58% (std 13.95%; min = 0%; max = 35%). Microsporidia were also readily detected with an estimated recovery of 46.81% overall (std 17.66%; min = 18%; max = 70%) for E. intestinalis and 38.90% (std 14.36%; min = 13%; max = 62%) for E. bieneusi.