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Applied and Environmental Microbiology, May 2003, p. 2842-2847, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2842-2847.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Recovery and Enumeration of Cryptosporidium parvum from Animal Fecal Matrices

Cheryl M. Davies,^1* Christine Kaucner,¹ Daniel Deere,² and Nicholas J. Ashbolt¹

Centre for Water and Waste Technology, School of Civil and Environmental Engineering, University of New South Wales, Sydney, New South Wales 2052,¹ Sydney Catchment Authority, Penrith, New South Wales 2751, Australia²

Received 9 September 2002/ Accepted 20 February 2003

Accurate quantification of Cryptosporidium parvum oocysts in animal fecal deposits on land is an essential starting point for estimating watershed C. parvum loads. Due to the general poor performance and variable recovery efficiency of existing enumeration methods, protocols were devised based on initial dispersion of oocysts from feces by vortexing in 2 mM tetrasodium pyrophosphate, followed by immunomagnetic separation. The protocols were validated by using an internal control seed preparation to determine the levels of oocyst recovery for a range of fecal types. The levels of recovery of 10² oocysts from cattle feces (0.5 g of processed feces) ranged from 31 to 46%, and the levels of recovery from sheep feces (0.25 g of processed feces) ranged from 21% to 35%. The within-sample coefficients of variation for the percentages of recovery from five replicates ranged from 10 to 50%. The ranges for levels of recovery of oocysts from cattle, kangaroo, pig, and sheep feces (juveniles and adults) collected in a subsequent watershed animal fecal survey were far wider than the ranges predicted by the validation data. Based on the use of an internal control added to each fecal sample, the levels of recovery ranged from 0 to 83% for cattle, from 4 to 62% for sheep, from 1 to 42% for pigs, and from 40 to 73% for kangaroos. Given the variation in the levels of recovery of oocysts from different fecal matrices, it is recommended that an internal control be added to at least one replicate of every fecal sample analyzed to determine the percentage of recovery. Depending on the animal type and based on the lowest approximate percentages of recovery, between 10 and 100 oocysts g of feces^-1 must be present to be detected.

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* Corresponding author. Mailing address: Centre for Water and Waste Technology, School of Civil and Environmental Engineering, University of New South Wales, Sydney, NSW 2052, Australia. Phone: 612 9385 5102. Fax: 612 9313 8624. E-mail: c.davies{at}unsw.edu.au.

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Applied and Environmental Microbiology, May 2003, p. 2842-2847, Vol. 69, No. 5
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.5.2842-2847.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Cox, P., Griffith, M., Angles, M., Deere, D., Ferguson, C. (2005). Concentrations of Pathogens and Indicators in Animal Feces in the Sydney Watershed. Appl. Environ. Microbiol. 71: 5929-5934 [Abstract] [Full Text]  
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