Location: Tick and Biting Fly Research
Project Number: 3094-32000-039-52-A
Project Type: Cooperative Agreement
Start Date: Jun 15, 2014
End Date: Oct 31, 2018
Genotype Babesia bovis, and as much as possible B. bigemina, isolates from blood and associated ticks infesting cattle in Puerto Rico.
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.