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ARS Home » Plains Area » Kerrville, Texas » Knipling-Bushland U.S. Livestock Insects Research Laboratory » LAPRU » Research » Publications at this Location » Publication #350370
Title: Efficient method for extracting DNA of parasites causing bovine babesiosis from tick vectors

Author
item GUERRA, JOSE - University Of Texas Rio Grande Valley
item Tidwell, Jason
item KLAFKE, GUIHERME - Department Of Energy
item Thomas, Donald
item PRUITT, KENNETH - University Of Texas Rio Grande Valley
item Miller, Robert
item Perez De Leon, Adalberto - Beto

Submitted to: Subtropical Agriculture and Environments
Publication Type: Other
Publication Acceptance Date: 2/9/2018
Publication Date: N/A
Citation: N/A

Interpretive Summary: Babesiosis is a disease of cattle caused by a parasite in the blood. It is spread among cattle by the bites of the cattle tick. The Cattle Fever Tick eradication program works to prevent the spread of this disease. In that effort the program tests blood and ticks taken from cattle exposed to the disease. The first step in the test is to extract the parasites DNA, if present, from the blood or the tick. The purpose of this research is to test three different methods of extracting the DNA from the southern cattle fever tick to determine if they are infected with the parasites causing bovine babesiosis. The factors used to measure the efficiency of each method are: quantity of DNA, purity of the DNA, and the time necessary to perform the extraction.

Technical Abstract: The southern cattle tick, Rhipicephalus (Boophilus) microplus, is an economically important pest costing animal agriculture billions of dollars worldwide. This research focuses on a comparison of three different tick DNA extraction methods: phenol-chloroform extraction (method 1), a modified version of the Aljanabi and Martinez protocol (method 2), and PureLink DNA Mini Kit protocol (method 3). These methods were used to purify nucleic acids and to eliminate contaminants to produce good quality DNA from R. microplus, which can be used to test for the presence of Babesia bovis, and B. bigemina. Efficacy is based on outcome factors such as purification quality, time efficiency, and average yield of quality DNA collected from each tick sample. With method 1, 85% of the samples produced quality DNA; 78% of the samples subjected to method 2 produced quality DNA; and, with method 3 75% of the samples produced quality DNA. Method 1 produced a higher yield of DNA whereas method 2 and 3 yielded equivalent DNA amounts. All methods yielded high quality DNA quality, yet methods 2 and 3 were more time efficient than method 1. This research highlights the importance of obtaining quality DNA for effective downstream analysis. In addition to testing for Babesia parasites, other molecular tests can be done on these samples for research on acaricide mutations, RT-PCR, population genetics, and next-gen sequencing.