Computer-Assisted Laser Scanning and Video Microscopy for Analysis of Cryptosporidium parvum Oocysts in Soil, Sediment, and Feces.

Applied and Environmental Microbiology
L J Anguish, W C Ghiorse

Abstract

A computer-assisted laser scanning microscope equipped for confocal laser scanning and color video microscopy was used to examine Cryptosporidium parvum oocysts in two agricultural soils, a barnyard sediment, and calf fecal samples. An agar smear technique was developed for enumerating oocysts in soil and barnyard sediment samples. Enhanced counting efficiency and sensitivity (detection limit, 5.2 x 10(sup2) oocysts(middot)g [dry weight](sup-1)) were achieved by using a semiautomatic counting procedure and confocal laser scanning microscopy to enumerate immunostained oocysts and fragments of oocysts in the barnyard sediment. An agarose-acridine orange mounting procedure was developed for high-resolution confocal optical sectioning of oocysts in soil. Stereo images of serial optical sections revealed the three-dimensional spatial relationships between immunostained oocysts and the acridine orange-stained soil matrix material. In these hydrated, pyrophosphate-dispersed soil preparations, oocysts were not found to be attached to soil particles. A fluorogenic dye permeability assay for oocyst viability (A. T. Campbell, L. J. Robertson, and H. V. Smith, Appl. Environ. Microbiol. 58:3488-3493, 1992) was modified by adding an immunost...Continue Reading

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Citations

May 7, 2003·Applied and Environmental Microbiology·Cheryl M DaviesNicholas J Ashbolt
Sep 6, 2003·Applied and Environmental Microbiology·W Quintero-BetancourtJ B Rose
Feb 16, 2002·Environmental Microbiology·S C KachlanyW C Ghiorse
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Jan 15, 2013·Parasitology Research·Jennifer A ZambriskiHussni O Mohammed
Jan 29, 2003·Journal of Environmental Quality·Xin Dai, Jan Boll
Jul 2, 1998·Applied and Environmental Microbiology·E Llobet-BrossaR Amann
Jul 2, 1999·Applied and Environmental Microbiology·E Kuczynska, D R Shelton
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Jan 20, 2018·Parasitology Research·Shahira A Ahmed, Panagiotis Karanis
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Jun 3, 1998·Applied and Environmental Microbiology·M J WalkerW C Ghiorse
Dec 5, 1998·Applied and Environmental Microbiology·L J RobertsonH V Smith
May 12, 2005·Environmental Science & Technology·Pamela L Young, Simeon J Komisar

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