• Parasites, Hosts and Diseases
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• v.32 no.1
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• pp.7-12
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• 1994

A calf and 50 mice were infected with Cryptosporidium parvum and their fecal materials were collected and treated 10 ether extraction (EE), followed by discontinuous sucrose gradients (DSG) method. EE method was to remove some of fat or lipid from feces. Sediments were washed by centrifugation ($1,500{\;}{\times}{\;}g$ for 10 min., 3 times) In phosphate-buffered saline and then these washed sediments were sleeved sequentially through stainless steel screens with a final mesh of 250 ($61{\;}{\mu\textrm{m}}$ porosity) to remove other debris. After sieving, the materials were suspended in 2.5% potassium dlchromate solution. Oocysts were counted by using a hemocytometer and the recovery rate of pure oocysts was calculated on the basis of the count. Following centrifugation ($1,500{\;}{\times}{\;}g$ for 30 min.) by DSG method, most oocysts were recovered at the interface between a gravity of 1.103 and 1.064. The recovery rates of pure oocysts from the fecal suspension of the calf ($3.8{\;}{\times}{\;}10^7/ml$) and the mouse ($3.2{\;}{\times}{\;}10^6/ml$) treated with EE method were 81.6% and 51.6%, respectively. It is suggested that the recovery rate was dependent on the number of oocysts In each suspension treated with EE method. To get the 50% recovery rate, there must be more than $2{\;}{\times}{\;}10^6$ oocysts per ml of the fecal suspension treated with EE method. By the combination of the two methods it was possible to isolate C. parvum oocysts from normal feces of the calf and mouse as well as from dlarrhelc feces.

• Parasites, Hosts and Diseases
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• v.41 no.4
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• pp.197-201
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• 2003

The present study was undertaken to determine the viability and infectivity of oocysts of Cryptosporidium baileyi that had been stored from 1 to 40 months at $4^{\circ}C$ preserved in 2.5% potassium dichromate solution. Oocysts of C. baileyi were purified from the feces of experimentally infected chickens using discontinuous sucrose gradients. Subsequently, the purified oocysts were suspended in 2.5% potassium dichromate solution at a concentration of $1{\;}{\times}10^7$ organism/ml, and their viabilities were assessed by nucleic acid staining, histologic examination, and infectivity to 2-day-old chickens. All chickens inoculated with oocysts that had been stored for 1-18 months developed patent infections, while chickens infected with older oocysts remained uninfected. Between 5.8% and 82.2% of the oocysts, stored at $4^{\circ}C$ in 2.5% potassium dichromate solution, were found to be viable, as determined by nucleic acid staining. Parasite colonization in the bursa of Fabricius was detected in the microvillus border of bursal epithelium. The finding that C. baileyi oocysts remain infective to chickens for at least 18 months offers important time-saving advantages to investigators who frequently require large numbers of oocysts.