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Investigation of Waterborne MicrosporidiaEPA Grant Number: U915584
Title: Investigation of Waterborne Microsporidia
Investigators: Mital, Jeffrey T.
Institution: John Jay College of Criminal Justice
EPA Project Officer: Broadway, Virginia
Project Period: September 1, 1999 through August 1, 2001
Project Amount: $60,444
RFA: STAR Graduate Fellowships (1999) RFA Text | Recipients Lists
Research Category: Fellowship - Environmental Science/Forensic Science , Academic Fellowships , Ecological Indicators/Assessment/Restoration
The objective of this research project is to develop a simple and efficient method for the routine detection of waterborne microsporidia, and to apply this method to the investigation of microsporidia in surface water.
The proposed technique concentrates microsporidia spores by filtration, and detects their presence using polymerase chain reaction (PCR) amplification of the gene that codes for a small subunit of ribosomal RNA in microsporidia. Submicron filters of different material, construction, and pore size are being tested to determine the most efficient filter for capture and recovery of spores. Various methods are being evaluated to determine the most effective means of extracting and isolating amplifiable DNA from within the tough spore. Primers are used to amplify DNA from most species of microsporidia that are infectious to humans. Surface water samples will be tested for the presence of microsporidia using the refined technique. The specific species present then will be determined by a combination of methods. Each species will result in an amplification product of a different length, ranging from 410 to 433 base pairs. The length of the PCR product can be determined, within one base pair, using Perkin Elmer's ABI Prism 310 Genetic Analyzer. Species confirmation will be accomplished by sequencing the PCR product.