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United States Department of Agriculture

Agricultural Research Service

Research Project: Genotyping Babesia Strains Circulating in Bovines and Cattle Fever Ticks in Puerto Rico

Location: Tick and Biting Fly Research

Project Number: 6205-32000-034-52
Project Type: General Cooperative Agreement

Start Date: Jun 15, 2014
End Date: Jun 14, 2015

Objective:
Genotype Babesia bovis, and as much as possible B. bigemina, isolates from blood and associated ticks infesting cattle in Puerto Rico.

Approach:
DNA will be extracted from blood samples and ticks from Puerto Rico and the samples screened by PCR for B. bovis and B. bigemina DNA in USDA-ARS lab. Babesia positive DNA samples will be forwarded to collaborator for genotyping. The 18S ribosomal RNA gene, which is the accepted standard genetic marker for molecular comparisons of Babesia (as well as other organisms), will be amplified by PCR and then cloned and sequenced by collaborator. For finer distinction among isolates, two markers will be analyzed. The genomic region spanning the ribosomal RNA internal transcribed spacers 1 and 2 and the 5.8S rRNA gene will be amplified by PCR, cloned, and sequenced. The surface antigen, MS2b, will be amplified and analyzed for restriction fragment length polymorphisms using HindIII, EcoR1, and PstI. Representative samples of the resulting MS2b genotypes will be sequence verified.

Last Modified: 9/10/2014
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