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Title: Molecular characterization of babesiosis infected cattle: Improvement of diagnosis and profiling of the immune response genes expression

Author
item ABDEL, AZIZ KB - Egypt National Research Center
item KHALIL, WKB - Egypt National Research Center
item MAHMOUD, MS - Egypt National Research Center
item HASAN, NA - Ain Shams University Of Cairo
item MABROUK, DM - Egypt National Research Center
item Suarez, Carlos

Submitted to: Global Veterinaria
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/1/2014
Publication Date: 2/1/2014
Citation: Abdel, A., Khalil, W., Mahmoud, M., Hasan, N., Mabrouk, D., Suarez, C.E. 2014. Molecular characterization of babesiosis infected cattle: Improvement of diagnosis and profiling of the immune response genes expression. Global Veterinaria. Global Veterinaria 12(2):197-206.

Interpretive Summary: The main aim of this study was to improve the detection of Babesia (B.) spp. in naturally infected cattle in Egypt. In addition, we analyzed the pattern of expression of selected cytokine genes in response to infection of bovines with B. bovis and B. bigemina. Infections were detected using both, traditional and novel diagnostic techniques competitive enzyme linked immunosorbent assay (cELISA) and nested polymerase chain reaction (nPCR). Blood samples were collected from 296 healthy Egyptian cattle and examined for babesial infection. Higher prevalence was recorded by cELISA, followed by nPCR, whereas lowest prevalence was recorded by microscopic examination of blood smears. Twenty bovine samples that were found positive for B. bovis, twenty samples that were positive for both species (B.bovis and B. bigemina) using cELISA and nPCR and twenty samples, negative for both species using the same techniques, were selected for the quantification of expression of the cytokine genes Interferon gamma (IFN-g), Interleukin-1 beta (IL-1b) and Transforming growth factor beta 1 (TGF-b1). All of the Babesia-positive samples were considered to have sub-clinical infection of babesiosis. The results revealed that infected cattle showed highly significant up-regulation of IL-1b and TGF-b1 genes and down-regulation of IFN-g gene when compared with non-Babesia infected animals. The improvement of babesial infection diagnosis combined with the understanding of the immune response will facilitate our understanding of the disease and the development of improved methods for control for these important diseases of cattle.

Technical Abstract: The main aim of this study was to improve the detection of Babesia (B.) spp. in naturally infected cattle in Egypt. In addition, we analyzed the pattern of expression of selected cytokine genes in response to infection of bovines with B. bovis and B. bigemina. Infections were detected using both, traditional and novel diagnostic techniques competitive enzyme linked immunosorbent assay (cELISA) and nested polymerase chain reaction (nPCR). Blood samples were collected from 296 healthy Egyptian cattle and examined for babesial infection. Higher prevalence was recorded by cELISA, followed by nPCR, whereas lowest prevalence was recorded by microscopic examination of blood smears. Twenty bovine samples that were found positive for B.bovis, twenty samples that were positive for both species (B.bovis and B. bigemina) using cELISA and nPCR and twenty samples, negative for both species using the same techniques, were selected to carry out a real-time PCR for the quantification of expression of the cytokines genes Interferon gamma (IFN-g), Interleukin-1 beta (IL-1b) and Transforming growth factor beta 1 (TGF-b1). All of the Babesia-positive samples were considered to have sub-clinical infection of babesiosis. The results revealed that infected cattle showed highly significant up-regulation of IL-1b and TGF-b1 genes and down-regulation of IFN-g gene when compared with non-Babesia infected animals. The improvement of babesial infection diagnosis combined with the understanding of the immune complex response will facilitate our understanding of the disease and the development of improved methods for control