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Applied and Environmental Microbiology, May 1999, p. 1998-2005, Vol. 65, No. 5
Department of
Microbiology,1 and Department of
Microbiology and Immunology, College of Veterinary
Medicine,2 Cornell University, Ithaca, New York,
and Environmental and Resource Sciences, University of Nevada,
Reno, Nevada3
Received 16 November 1998/Accepted 2 February 1999
A small-volume sentinel chamber was developed to assess the effects
of environmental stresses on survival of sucrose-Percoll-purified Cryptosporidium parvum oocysts in soil and animal wastes.
Chambers were tested for their ability to equilibrate with external
chemical and moisture conditions. Sentinel oocysts were then exposed to stresses of the external environment that affected their viability (potential infectivity), as indicated by results of a dye permeability assay. Preliminary laboratory experiments indicated that temperatures between 35 and 50°C and decreases in soil water potential (
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Use of a Sentinel System for Field Measurements of
Cryptosporidium parvum Oocyst Inactivation in Soil and
Animal Waste
0.003 to
3.20 MPa) increased oocyst inactivation rates. The effects of two
common animal waste management practices on oocyst survival were
investigated on three dairy farms in Delaware County, N.Y., within the
New York City watershed: (i) piling wastes from dairy youngstock
(including neonatal calves) and (ii) spreading wastes as a soil
amendment on an agricultural field. Sentinel containers filled with
air-dried and sieved (2-mm mesh) youngstock waste or field soil were
wetted and inoculated with 2 million oocysts in an aqueous suspension
and then placed in waste piles on two different farms and in soil
within a cropped field on one farm. Controls consisted of purified
oocysts in either phosphate-buffered saline or distilled water
contained in sealed microcentrifuge tubes. Two microdata loggers
recorded the ambient temperature at each field site. Sentinel
experiments were conducted during the fall and winter (1996 to 1997)
and winter (1998). Sentinel containers and controls were removed at 2- to 4-week intervals, and oocysts were extracted and tested by the dye
permeability assay. The proportions of potentially infective oocysts
exposed to the soil and waste pile material decreased more rapidly than their counterpart controls exposed to buffer or water, indicating that
factors other than temperature affected oocyst inactivation in the
waste piles and soil. The effect of soil freeze-thaw cycles was evident
in the large proportion of empty sentinel oocysts. The potentially
infective sentinel oocysts were reduced to <1% while the proportions
in controls did not decrease below 50% potentially infective during
the first field experiment. Microscopic observations of empty oocyst
fragments indicated that abrasive effects of soil particles were a
factor in oocyst inactivation. A similar pattern was observed in a
second field experiment at the same site.
*
Corresponding author. Mailing address: Department of
Microbiology, Wing Hall, Cornell University, Ithaca, NY 14853-8101. Phone: (607) 254-5117. Fax: (607) 255-3904. E-mail:
mbj1{at}cornell.edu.
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