Cryptosporidium in Drinking Water Racheal Nell Title: Cryptosporidium spp. in Drinking Water: Samples from rural sites in Switzerland Introduction: Cryptosporidium is a coccidian parasite that infects the intestine of warm blooded animals and humans. Crypto's life cycle first starts in contaminated matter, Crypto then feeds off of the fecal matter and waits for the next host. They can survive several months without a host, a host is infected orally threw direct fecal contact or contaminated food and water. Once they are directly put into the stomach. Most parasites would be killed by the toxic acids found in mammal’s stomachs but Crypto's niche of having a protective double layer oocysts around it. Crypto is then passes through the stomach into the intestine. In the intestine Crypto latches on to the intestinal lining and the new Crypto sporozoites is released. The sporozoites then invade the epithelial cells and start to reproduce asexually or sexually within the cell. Once the epithelial cell is full of sporozoites the cell wall breaks and they are released into the intestine. They then try to reattach themselves to the lining or is released from the host and the cycle continues. When infected the host will experience symptoms 1-14 days after being infected. severe diarrhea, dehydration, cramps, fever and nausea are some of the common symptoms that lead into weight loss and in some extreme cases death. Reason for research: In 1995 a Cryptosporidium outbreak occurred in Milwaukee Japan, infecting 15,000-400,000 people. Soon after this outbreak multiple investigations developed to find the source of the contamination. It was concluded that the source of the infection was from drinking water that was contaminated with fecal material. Nowadays, we have developed regulations to keep Crypto under control reducing the infection rate to 1 for every 10,000. In Switzerland, the drinking water distribution system include and extra treatment to insure the water in Crypto free. However, in the rural areas of Switzerland the water is often in contact with grazing animal's excrement and feces. The water is rarely treated but depended on for consumption, the aim of this study was to see what the risk of infection would be through consumption of the rural drinking water. Method & Materials: Three samples of water (100mL) was taken from three different locations of rural areas, site A- Alpine Village, Site B- Alpine Camp and Site C Mittelland town in Switzerland at three different times. The samples were then transferred to a lab and tested for Cryptosporidium spp and E.coli. For detection of the Crypto, the oocysts were stained with specific surface antibodies and then looked at one by one through fluorescence microscopy. The oocysts were then suspended my lysed open for DNA extraction. The DNA extraction was then used for PCR and sequencing of the PCR products for further genotyping. The PCR products were then put through sequence alignments for phylogenic analysis. Results: The 18SrRNA gene of Crypto sp. was collected from a water sample at site A and deposited into GenBank. At Site A, all three samples were contaminated with both Crypto and E.coli. At site B the samples varied and had the highest concentration of oocyst after a heavy rainfall. At Site C only one sample had an elevated oocyst concentration. Discussion/Conclusion: After the experiment it was determined that the concentrations of the oocyst present in the total water samples could be the direct source of the outbreak in the children. They found a direct correlation with the amount of E.coli and Crypto, the greater concentration of E.coli increased the concentration of Crypto oocsyt. Verifying that the contamination could be coming from the fecal matter from all of the cattle and land animals. When the heavy rain fell they noticed a great increase of concentration of oocyst, the soil better infiltrates the manure during the heavy rainfall but on dry land the manure stays on the surface of the soil. Bibliography: http://www.ncbi.nlm.nih.gov/pubmed/23037557