Non-invasive methods for identifying oocysts of Sarcocystis spp. from definitive hosts |
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| Authors: | Zheng Xiang Xinwen Chen Lijun Yang Yongshu He Runsheng Jiang Benjamin M Rosenthal Pengtao Luan SW Attwood Yangxian Zuo Ya-ping Zhang Zhaoqing Yang |
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| Institution: | 1. Parasitology Department, Kunming Medical College, Kunming, Yunnan 650031, PR China;2. Department of Biology, Yunnan University, Kunming, Yunnan 650091, PR China;3. Division of Health Social Sciences, Institute for Health Science, Kunming Medical College, Kunming, Yunnan 650031, PR China;4. Animal Parasitic Disease Laboratory, Agricultural Research Service, US Department of Agriculture, BARC East Building, 1180 Beltsville, MD 20705, USA;5. State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, PR China;6. Laboratory for Conservation and Utilization of Bioresource, Yunnan University, Kunming 650091, PR China;7. State Key Laboratory of Genetic Resource and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, PR China;1. Animal Parasitic Diseases Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, United States Department of Agriculture, Beltsville 20705-2350, MD, USA;2. Alaska Department of Fish and Game, Fairbanks, AK 99701, USA;3. United States Department of Agriculture, Animal and Plant Health Inspection Service, Wildlife Services, 9001 East Frontage Road, Suite A, Palmer 99645, AK, USA;4. Veterinary Pathology Services, Joint Pathology Center, 606 Stephen Sitter Avenue, Silver Spring 20910, MD, USA;1. Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH, 43210, United States;2. Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH, 43210, United States;3. Department of Veterinary Science, University of Kentucky, 108 Gluck Equine Research Center, Lexington, KY, 40546, United States;4. Rood & Riddle, Equine Hospital, Lexington, KY, 40511, United States;1. Department of Food Safety and Infection Biology, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, P.O. Box 8146 Dep., NO-0033 Oslo, Norway;2. Norwegian Veterinary Institute, P.O. Box 750 Sentrum, NO-0106 Oslo, Norway;1. Parasitology Division, National Veterinary Research Institute (NVRI), PMB 01 Vom, Plateau State, Nigeria;2. Koret School of Veterinary Medicine, The Hebrew University of Jerusalem, Rehovot, Israel;3. Parasitology Unit, Department of Microbiology and Molecular Genetics, The Kuvin Center for the Study of Infectious and Tropical Diseases, Hebrew University-Hadassah Medical School, Jerusalem, Israel;1. Department of Preventive Veterinary Medicine and Animal Health, University of Sao Paulo, Sao Paulo, Sao Paulo, Brazil;2. Department of Microbiology and Parasitology, University of Pelotas, Pelotas, Rio Grande do Sul, Brazil;1. Laboratory of Veterinary Pathology, Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium;2. Department of Veterinary Public Health and Food Safety, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium;3. Laboratory of Parasitology, Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium;4. Center for Medical Genetics Ghent (CMGG), Ghent University Hospital, De Pintelaan 185, 9000 Ghent, Belgium |
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| Abstract: | Because the excreted sporocysts and/or oocysts of various species of Sarcocystis may not be discriminated morphologically, we sought to validate a diagnostic technique based on variation in the 18S rDNA sequence. Oocysts and/or sporocysts from three taxa of Sarcocystis were collected from human, feline, and canine definitive hosts that had fed upon meats infected with the muscle cysts of Sarcocystis hominis, Sarcocystis fusiformis and a species of Sarcocystis from water buffalo that could not be distinguished from Sarcocystis cruzi. Using a new collection method employing filter paper, these excreted oocysts and sporocysts were subjected to DNA extraction, as were the corresponding muscle cysts. Methods employing PCR–RFLP and DNA sequencing of a partial 18S rDNA gene (ssrRNA) sequence were then used to successfully distinguish among the three taxa. The same, unique restriction digestion pattern characterizes the tissue cysts and oocysts and/or sporocysts of each parasite taxon. The technique makes possible amplification and identification of species specific gene sequences based on DNA extracted from as few as 7 excreted sporocysts (the equivalent of 3 and 1/2 oocysts) from freshly prepared material, or as few as 50 sporocysts from feces samples that had been stored in potassium dichromate (K2Cr2O7) for as long as 6 years. This represents the first report using molecular diagnostic procedures to diagnose oocysts of Sarcocystis in faecal samples, describing a valuable new tool for studying the epidemiology of various Sarcocystis species. |
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| Keywords: | Sarcocystis Oocysts Faeces Taxonomy 18S rRNA Diagnostics |
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