Rivera-Diaz, Natalia A.

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    Use of the real time TAQMAN®-PCR for detection of Babesia bovis and Babesia bigemina infection in dairy farms of Puerto Rico
    (2017) Rivera-Diaz, Natalia A.; Mesonero, Alexander; College of Agricultural Sciences; Jimenez, Esbal; Sánchez, Héctor; Klafke, Guilherme; Department of Animal Science; Resto Irizarry, Pedro J.
    The dairy industry constitutes an important resource to the socioeconomically development of Puerto Rico. It is the main agricultural enterprise supporting more than 25,000 employees and producing nearly $215 million during the fiscal year 2010 (26% of the total agriculture revenue. However, the production and stability of our dairy industry is challenged by hemoparasitic diseases like babesiosis, a vector borne disease transmitted by the cattle fever tick (Rhipicephalus microplus), that result in economical and health repercussions to the dairy farms. Fast and efficient identification of cattle babesiosis can prevent or decrease the dissemination of the disease, and consequently allow for early control and proper treatment. The present study evaluates the efficiency of a multiplex Real Time Taqman®- PCR technique as a laboratory diagnostic test for cattle babesiosis, compared with conventional detection techniques currently used (immunoassays-ELISA; conventional end point PCR). The prevalence of Babesia bovis and B.bigemina in different cattle ranches was analyzed taking into account the results obtained with nested-PCR and the suggested real time Taqman® PCR. Sample collection was performed in four commercial dairy cattle farms located in the northwest and southwest part of Puerto Rico. A total of 158 cows were tested for identification of Babesia bovis and Babesia bigemina using nPCR and real time Taqman®-PCR. A McNemar’s Chi-square test performed with the SAS/STAT program indicated that both detection methods resulted with 100% of specificity with some variation of sensitivity for detection of B.bovis and B.bigemina. Statistical analysis revealed no difference between tests, with pvalue 0.0833 for B.bovis and pvalue 0.3173 for B.bigemina (p value>0.05). Despite the lack of significant differences in the results obtained between the two modalities employed, there are clear advantages of using Real Time Taqman® PCR over nPCR. Time consumption in preparation and turnaround time using Real Time Taqman®-PCR, especially if a large amount of samples are to be processed, demonstrated to be much lower than nPCR. Results obtained in the study indicated the prevalence of Babesia bovis in the premises under evaluation, a total of 15 positive results. San Sebastian farm being the main premise infected with a total of 12 positive results for Babesia bovis.