| ObjectiveCryptosporidium is an opportunistic protozan parasite that is widely distributed in the world. It was emerged as one of six pathogens causing humans diarrhea. Cryptosporidium can cause waterborne or food - borne diarrhea outbreak, it is also the common reason, which causes the death of AIDS patients and children.Cryptosporidium has been one of important pathogenic testing items for AIDS overseas. The features of the oocysts is useful in diagnosis for the Crypto-sporidiosis. The morphology of Cryptosporidium oocysthas been described, but, there is no any report in the morphological parameters about Cryptoridium by digital image processing technology as well as no any comparative report in oocysts from different area and different animal by this technology. The morphological parameters achieved by computer is more objective, precise, sensitive and massive than the traditional methods in obtaining morphologic information. Recently, the technology has been used on researching in some pathogen, such as Pneumocystis. To define the morphological changing feature and offer more confident morphologic parameters for morphological studying, the morphological parameter of Cryptosporidium oocyst was established and analyzed by digital imagine processing technologyMethod1. The source of Cryptosporidium oocysts1.1 Muris oocysts were collected from feces of Shen - Yang KM mice from experiment animal department of China Medical University, immunodepressed by Dexamethsone for 10 days.1.2 bovine oocysts were collected from Chang - Chun catties ( Provided by Prof, zhang xi chen).2. Staining and definiting of Cryptosporidium oocystsFeces of positive Cryptosporidium oocyst from KM mice was diluted and smeared by normal saline, smears were stained by modified fast - acid staining and mounted for using.3. Geting and analysis images of Cryptosporidium oocystsOocysts'images were acquired by photograph system, oocyst length, oocyst width, perimeter, area and equivalent diameter etc. were obtained by computer digital image processing system.4. Data analysisThe oocyst morphological parameters were analyzed by Excell statistic analysis software and t test.Result1. The inducing result of mouse infected by CryptosporidiumThe onset of oocyst shedding occurred on 5 — day after inducing, severe infection was seen on 6 to 7 - day. Symptoms such as depression and reducing appetite and water occured at the same time. All mice infected Cryptosporidium were killed on 10 - day. Feces obtained from intestine was stained by modified fast — acid staining and examined under microscope, Cryptosporidium oocysys were full of visual fields.2. Staining feature of Cryptosporidium oocystsThe Cryptosporidium oocysts stained by fast - acid staining appeared as round densely stained red bodies on a blue - green background. The wall is smooth and colorless, crescent sporozoites and residual granules ( stained brown or black) are clear, a vacuoles in the body is also observed.3. Collecting oocysts'images and analyzing data1190 muris Cryptosporidium oocysts were collected. The average of oocyst length is 5. 93|xm, ranging from 3. 36jxm ~ 8. 51|xm in 95% confidence interval. The average of oocyst width is 4.96|am, ranging from 3.26juim ~ 6. 66u,m in 95% confidence interval. The average of perimeter is 18. 03 (im, the average of area is 16.08|xm2. The average of shape index is 1.19.Collecting bovine Cryptosporidium oocysts 922. The average of oocyst length is 5. 19[im, ranging from 2. 91|xm ~7. 47(xm in 95% confidence interval. The average of oocyst width is 4.17 juun, ranging from 2. 72jmm ~5. 36juum in 95% confidence interval. The average of perimeter is 15.74(xm, the average of area is 12. lljxm2. The average of shape index is 1. 25.4. Comparing of oocysts comimg from different area and animalsThe two host - source Cryptosporidium oocysts are different significantly in length and width (p < 0.01) byt -test,but the shape index is similar. Oocysts from Shen -Yang KM mice are bigger than those from Chang - Chun catties by statistic analysis.Conclusion1. The morphological parameters obtain by digital image processing technology from muris and bovine Cryptosporidium oocysts are difference by t test.2. The data and parameters of oocysts obtained from computer are objective and precise, offering scientific foundation for measuring and evaluating oocysts as well as taxonomy of Cryptosporidium spp.
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