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A new intestinal trypanosomatid was found parasitizing Megaselia scalaris. Forms with posterior kinetoplasts occurred in both natural infections and cloned cultures, but not in experimental infections. Average size and morphological variation were reduced in experimental infections with cloned stocks, compared to natural infections. A growth comparison was made between Herpetomonas muscarum and H. megaseliae. Differences in day of peak population, maximum numbers, and total time of survival were observed in culture. Attempts to infect Drosophila melanogaster with H. megaseliae were unsuccessful.