Genetic Characterization and Phylogenetic Analysis of <i>Babesia bigemina</i> Isolates in Cattle from South Africa Based on <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> Genes
<i>Babesia bigemina</i> is one of the most important etiological agents of bovine babesiosis, a tick-borne disease posing a major threat in the livestock industry globally, including South Africa. Despite the huge economic impact of cattle babesiosis in South Africa, antigenic variation...
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| author | Phillip Senzo Mtshali Moses Sibusiso Mtshali |
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| description | <i>Babesia bigemina</i> is one of the most important etiological agents of bovine babesiosis, a tick-borne disease posing a major threat in the livestock industry globally, including South Africa. Despite the huge economic impact of cattle babesiosis in South Africa, antigenic variation observed among <i>B. bigemina</i> strains worldwide has impeded the successful development of a single vaccine with the potential to eliminate the disease. As such, there is still a dearth of information regarding the conservation of <i>B. bigemina</i> genes encoding functionally important proteins that play a crucial role during the invasion of bovine erythrocytes by merozoites. Fifty blood samples previously collected from cattle in eight provinces of South Africa were genetically tested for the presence of <i>B. bigemina</i> DNA fragments using four nested PCR-based assays. The genes targeted coded for <i>Spe</i>I-<i>Ava</i>I restriction fragment, rhoptry-associated protein 1 (<i>BgRAP-1</i>), apical membrane antigen 1 (<i>BgAMA-1</i>) and β-tubulin (<i>BgβTUB</i>). PCR-generated fragments of randomly selected samples were sequenced. BLAST searches in GenBank were performed with newly determined sequences to search for homologous sequences. Neighbor-joining phylogenies were inferred from aligned, contiguous sequences of <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> genes. Nested PCR assays generated single fragments of 170 bp, 472 bp, 765 bp and 302 bp for <i>Spe</i>I-<i>Ava</i>I, <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> fragments, respectively. Of the 50 bovine samples tested by nested PCR, 82% (42/50; 95% CI = 69.2–90.2%), 68% (34/50; 95% CI = 54.2–79.2%), 50% (25/50; 95% CI = 36.6–63.4%) and 46% (23/50; 95% CI = 33.0–59.6%) possessed <i>B. bigemina</i>-specific <i>Spe</i>I-<i>Ava</i>I, <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> DNA fragments, respectively. The <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> sequences of South African <i>B. bigemina</i> isolates shared 98–100% similarity with previously reported sequences of strains originating from cattle in countries other than South Africa. The high genetic conservation observed among geographical isolates of <i>B. bigemina</i> suggests the conserved functional role of BgRAP-1 and BgAMA-1 proteins as potential candidates that could be incorporated in recombinant subunit vaccines. |
| format | Article |
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| spelling | doaj-art-2e531f52f2f2453aa69c9d0e91c672ce2025-08-20T02:24:43ZengMDPI AGBiology2079-77372025-03-0114435510.3390/biology14040355Genetic Characterization and Phylogenetic Analysis of <i>Babesia bigemina</i> Isolates in Cattle from South Africa Based on <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> GenesPhillip Senzo Mtshali0Moses Sibusiso Mtshali1Veterinary Parasitology Programme, Research and Scientific Services Department, National Zoological Gardens of South Africa, Pretoria 0001, South AfricaVeterinary Parasitology Programme, Research and Scientific Services Department, National Zoological Gardens of South Africa, Pretoria 0001, South Africa<i>Babesia bigemina</i> is one of the most important etiological agents of bovine babesiosis, a tick-borne disease posing a major threat in the livestock industry globally, including South Africa. Despite the huge economic impact of cattle babesiosis in South Africa, antigenic variation observed among <i>B. bigemina</i> strains worldwide has impeded the successful development of a single vaccine with the potential to eliminate the disease. As such, there is still a dearth of information regarding the conservation of <i>B. bigemina</i> genes encoding functionally important proteins that play a crucial role during the invasion of bovine erythrocytes by merozoites. Fifty blood samples previously collected from cattle in eight provinces of South Africa were genetically tested for the presence of <i>B. bigemina</i> DNA fragments using four nested PCR-based assays. The genes targeted coded for <i>Spe</i>I-<i>Ava</i>I restriction fragment, rhoptry-associated protein 1 (<i>BgRAP-1</i>), apical membrane antigen 1 (<i>BgAMA-1</i>) and β-tubulin (<i>BgβTUB</i>). PCR-generated fragments of randomly selected samples were sequenced. BLAST searches in GenBank were performed with newly determined sequences to search for homologous sequences. Neighbor-joining phylogenies were inferred from aligned, contiguous sequences of <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> genes. Nested PCR assays generated single fragments of 170 bp, 472 bp, 765 bp and 302 bp for <i>Spe</i>I-<i>Ava</i>I, <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> fragments, respectively. Of the 50 bovine samples tested by nested PCR, 82% (42/50; 95% CI = 69.2–90.2%), 68% (34/50; 95% CI = 54.2–79.2%), 50% (25/50; 95% CI = 36.6–63.4%) and 46% (23/50; 95% CI = 33.0–59.6%) possessed <i>B. bigemina</i>-specific <i>Spe</i>I-<i>Ava</i>I, <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> DNA fragments, respectively. The <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> sequences of South African <i>B. bigemina</i> isolates shared 98–100% similarity with previously reported sequences of strains originating from cattle in countries other than South Africa. The high genetic conservation observed among geographical isolates of <i>B. bigemina</i> suggests the conserved functional role of BgRAP-1 and BgAMA-1 proteins as potential candidates that could be incorporated in recombinant subunit vaccines.https://www.mdpi.com/2079-7737/14/4/355<i>Babesia bigemina</i>cattleSouth Africagenetic characterizationphylogenies |
| spellingShingle | Phillip Senzo Mtshali Moses Sibusiso Mtshali Genetic Characterization and Phylogenetic Analysis of <i>Babesia bigemina</i> Isolates in Cattle from South Africa Based on <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> Genes Biology <i>Babesia bigemina</i> cattle South Africa genetic characterization phylogenies |
| title | Genetic Characterization and Phylogenetic Analysis of <i>Babesia bigemina</i> Isolates in Cattle from South Africa Based on <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> Genes |
| title_full | Genetic Characterization and Phylogenetic Analysis of <i>Babesia bigemina</i> Isolates in Cattle from South Africa Based on <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> Genes |
| title_fullStr | Genetic Characterization and Phylogenetic Analysis of <i>Babesia bigemina</i> Isolates in Cattle from South Africa Based on <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> Genes |
| title_full_unstemmed | Genetic Characterization and Phylogenetic Analysis of <i>Babesia bigemina</i> Isolates in Cattle from South Africa Based on <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> Genes |
| title_short | Genetic Characterization and Phylogenetic Analysis of <i>Babesia bigemina</i> Isolates in Cattle from South Africa Based on <i>BgRAP-1</i>, <i>BgAMA-1</i> and <i>BgβTUB</i> Genes |
| title_sort | genetic characterization and phylogenetic analysis of i babesia bigemina i isolates in cattle from south africa based on i bgrap 1 i i bgama 1 i and i bgβtub i genes |
| topic | <i>Babesia bigemina</i> cattle South Africa genetic characterization phylogenies |
| url | https://www.mdpi.com/2079-7737/14/4/355 |
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