Submitted to: American Association of Veterinary Parasitologists Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 5/14/2003
Publication Date: 8/1/2003
Citation: Trout, J.M., Santin, M., Fayer, R. 2003. Genotype of giardia in dairy calves and pastureland wildlife. American Association of Veterinary Parasitologists Proceedings, July 19-22, 2003, Denver, CO. p.47.
Technical Abstract: Fecal samples were collected from 2-6 week-old dairy calves from two farms in each of the following states: VT, NY, PA, MD, VA, NC, and FL, as well as from white-tailed deer in MD and coyote in PA. For analysis by immunofluorescent microscopy and PCR, fifteen gram portions of each sample were mixed with water, passed through a 45 micron screen, mixed with CsCl, and centrifuged. The top 4 ml of suspension from the tubes was collected, pelleted by centirfugation, washed with distilled water, and resuspended in 100 :l of distilled water. Fifty :l was stained with a commercial Giardia specific mAb. The remianing 50 :l was subjected to DNA extraction using a commercial tissue DNA extraction kit. For coyotes, duodenal scrapings were also obtained. A portion was subjected to DNA extraction as described above, while the remainder was fixed and examined by light microscopy for the the presence of trophozoites. PCR amplification was done for the TPI, 16S, and $-giardin genes, using a nested PCR for TPI and 16S. PCR products were sequenced and the Giardia gene sequences obtained from animals in this survey were compared to each other and to previously sequenced Giardia isolates. PCR and gene sequence analysis revealed that calves were infected primarily with the hoofed-livestock genotype, however, a small percentage of Assemblage A genotype was also detected. Assemblage A Giardia was also detected in one white-tailed deer. Preliminary sequencing of the 16s gene suggests the cyote Giardia may also fall into Assemblage A. MEETING ABSTRACT: AMERICAN ASSOCIATION OF VETERINARY PARASITOLOGISTS 48TH ANNUAL CONFERENCE