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The diagnostic performance of a cocktail formula consisting of two Babesia (B.) bovis recombinant proteins, including spherical body protein 1 (BbSBP-1) and spherical body protein 4 (BbSBP-4), was evaluated in the present study for the global detection of B. bovis infection in cattle and for the differentiation between B. bovis and B. bigemina infections. The efficacy and the practicality of the rBbSBP-1 and rBbSBP-4 cocktail formula for differentiation between the infection caused by both parasites were assessed using indirect enzyme-linked immunosorbent assay (iELISA) with serum samples collected from cattle experimentally infected by B. bovis (n = 33) or B. bigemina (n = 30). Cocktail antigen exhibited the highest optical density (OD) values with B. bovis-infected sera and the lowest OD values with normal bovine sera or B. bigemina-infected sera in comparison with the single antigen. A total of 581 field serum samples collected from four countries with known B. bovis endemicity: Ghana (n = 154), Egypt (n = 162), Thailand (n = 96), and South Africa (n = 169) were screened also in the current study using iELISA and the results were compared to those of indirect fluorescent antibody test (IFAT) as a reference. A cocktail formula (rBbSBP-1 and rBbSBP-4) exhibited the highest concordance rate (89.90%) and kappa value (0.73). The obtained results revealed the reliability of the rBbSBP-1 and rBbSBP-4 cocktail antigen for the detection of specific antibodies to B. bovis in cattle and demonstrated the usefulness of cocktail antigen for differentiation between B. bovis and B. bigemina infections compared with the single antigen in cattle, which will be useful for epidemiological surveys and control of bovine babesiosis., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Babesia bovis, B. bigemina, Trypanosoma evansi, and Anaplasma marginale infections cause serious diseases in cattle, and are primarily transmitted by arthropod vectors (ticks for B. bovis, B. bigemina, and A. marginale and various types of flies for T. evansi). In the last few years, there have been many reports of a high prevalence of certain protozoan infections in northern Egypt, but no accurate or adequate data are available for the southern regions. Therefore, in this study, we screened for evidence of such diseases in economically important cattle species using serum samples. The seroprevalence of protozoan infections in cattle was determined with enzyme-linked immunosorbent assays using species-specific diagnostic antigens. In a total of 301 cattle serum samples, 27 (9.0%), 100 (33.2%), and 127 (42.2%) were positive for specific antibodies against B. bovis, B. bigemina, and T. evansi, respectively. Sera from 90 cattle were also tested for antibodies against A. marginale, and 25 (28%) of them were positive. The highest coinfection rate occurred for B. bigemina and T. evansi with 10.6% (32/301). When age, sex, locality, and breeding system were investigated as predisposing factors, bulls and cattle <3 years old were more vulnerable to B. bovis infections than older animals, and geographic location affected the B. bigemina infection rate. The recorded seroprevalence of hemoprotozoan parasites and A. marginale in cattle suggests that these diseases have the potential capacity to detrimentally affect meat and milk production in southern Egypt., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Cattle, buffaloes, and sheep are the main sources of meat and milk in Egypt, but their productivity is thought to be greatly reduced by hemoprotozoan parasitic diseases. In this study, we analyzed the infection rates of Babesia bovis, Babesia bigemina, Theileria annulata, and Theileria orientalis, using parasite-specific PCR assays in blood-DNA samples sourced from cattle (n=439), buffaloes (n=50), and sheep (n=105) reared in Menoufia, Behera, Giza, and Sohag provinces of Egypt. In cattle, the positive rates of B. bovis, B. bigemina, T. annulata, and T. orientalis were 3.18%, 7.97%, 9.56%, and 0.68%, respectively. On the other hand, B. bovis and T. orientalis were the only parasites detected in buffaloes and each of these parasites was only found in two individual DNA samples (both 2%), while one (0.95%) and two (1.90%) of the sheep samples were positive for B. bovis and B. bigemina, respectively. Sequence analysis showed that the B. bovis Rhoptry Associated Protein-1 and the B. bigemina Apical Membrane Antigen-1 genes were highly conserved among the samples, with 99.3-100% and 95.3-100% sequence identity values, respectively. In contrast, the Egyptian T. annulata merozoite surface antigen-1 gene sequences were relatively diverse (87.8-100% identity values), dispersing themselves across several clades in the phylogenetic tree containing sequences from other countries. Additionally, the T. orientalis Major Piroplasm Surface Protein (MPSP) gene sequences were classified as types 1 and 2. This is the first report of T. orientalis in Egypt, and of type 2 MPSP in buffaloes. Detection of MPSP type 2, which is considered a relatively virulent genotype, suggests that T. orientalis infection may have veterinary and economic significance in Egypt. In conclusion, the present study, which analyzed multiple species of Babesia and Theileria parasites in different livestock animals, may shed an additional light on the epidemiology of hemoprotozoan parasites in Egypt., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)