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DETECTION OF CRYPTOSPORIDIUM PARVUM AND GIARDIA LAMBLIA CARRIED BY SYNANTHROPIC FLIES BY COMBINED FLUORESCENT IN SITU HYBRIDIZATION AND A MONOCLONAL ANTIBODY

Wild-caught synanthropic flies were tested for the presence of Cryptosporidium parvum and Giardia lamblia on their exoskeletons and in their digestive tracks by fluorescent in situ hybridization and fluorescein isothiocyanate (FITC)–conjugated monoclonal antibody (MAb) against Cryptosporidium and Giardia cell wall epitopes. The levels of C. parvum were positively correlated with the levels of G. lamblia, indicating a common source of contamination. The majority of oocysts and cysts were potentially viable (C. parvum = 80% and G. lamblia = 69%). More G. lamblia cysts occurred on the exoskeleton of the flies than within the digestive tracts; the opposite relationship was observed for C. parvum. No genotype other than C. parvum G2 was found to be associated with flies. Because filth flies carry viable C. parvum oocysts and G. lamblia cysts acquired naturally from unhygienic sources, they can be involved in the epidemiology of cryptosporidiosis and giardiasis. Fluorescent oligonucleotide probes used together with FITC–conjugated MAb represent a convenient and cost-effective technique for rapid and specific identification of human-infectious species of Cryptosporidium and Giardia mechanically transported by flies, and for the assessment of the viability of these pathogens.

Received December 27, 2001. Accepted for publication May 9, 2002.

Acknowledgments: We thank D. E. Griffin (Johns Hopkins University, Baltimore, MD) and J. Trout (United States Department of Agriculture, Beltsville, MD) for facilitating this study, and D. Norris (Johns Hopkins University, Baltimore, MD) for his comments on the manuscript. We also thank B. Engber (North Carolina Department of Environment and Natural Resources, Raleigh, NC) for his consultation on capturing of wild flies and selection of capture sites.

Financial support: The study was supported by the U.S. Environmental Protection Agency (Washington, DC) (grant no. R824995), the Faculty Innovation Fund (grant no. H040-951-0189), The Center for A Livable Future (Baltimore, MD) (grant no. H040-951-0180), and the AKC Fund (New York, NY) (grant no. H630-951-2002).

Authors’ addresses: Thaddeus K. Graczyk and Ronald Knight, The W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, 615 North Wolfe Street, Baltimore, MD 21205. Barbara H. Grimes, Division of Environmental Health, On-Site Wastewater Section, 1642 Mail Service Center, Raleigh, NC, Telephone: 919-715-0141, Fax: 919-715-3227. Alexandre J. Da Silva and Norman J. Pieniazek, Division of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30341, Telephone: 770-488-4073, Fax: 770-488-4108. Duncan A. Veal, Fluorometrics Research Team, Department of Biologic Sciences, Macquarie University, Sydney, New South Wales 2109, Australia, Telephone: 61-2-9850-8185, Fax: 61-2-9850-8253.

Reprint requests: Thaddeus K. Graczyk, The W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, 615 North Wolfe Street, Baltimore, MD 21205, Telephone: 410-614-4984, Fax: 410-955-0105, E-mail: tgraczyk{at}jhsph.edu.

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