Unique cultural methods used to detect viable cryptosporidium parvum oocysts in environmental samples

Cryptosporidium parvum is an infectious enteric protozoan parasite that causes waterborne disease, severe gastroenteritis and is associated with high mortality in immunocompromised individuals. Detection of oocysts in water is very difficult and current methodologies do not determine viability. This project has focused on low level detection of Cryptosporidium parvum in environmental samples using a unique cultural method. Previously, cell culture methods have been used to assess the developmental stages of Cryptosporidium; however, no cultural methods have been employed with environmental samples. The percentage of viable oocysts can be estimated by detecting intracellular developmental stages of the parasite using fluorescently labelled antibodies. Other methods are not capable of low level detection or high sensitivity. We are evaluating detection of single foci of infection, indicating that one of the four sporozoites released from the viable oocyst has infected a single cell.