Multi-locus typing scheme for Babesia bovis and Babesia bigemina reveals high levels of genetic variability in strains from Northern Argentina

Guillemi, Eliana Carolina; Ruybal, Paula; Lia, Verónica Viviana; González, Sergio; Farber, Marisa Diana; Wilkowsky, Silvina Elizabeth

doi:10.1016/j.meegid.2012.12.005

Cited by 13 publications

(10 citation statements)

References 44 publications

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“…Therefore, special management measures have to be taken to prevent economic losses related to infection of this pathogen in cattle populations such as decreasing the tick transfer between cattle and buffaloes on farms with both hosts, as well as sharing of needles between them. On the other hand, the lack of genetic structure could be due to a variability of apicomplexan parasites that has been proposed as a survival strategy against the vertebrate immune response [ 44 – 46 ]. In this sense, genetic diversity observed here could be associated with continuous immune pressure by both hosts.…”

Section: Discussionmentioning

confidence: 99%

Molecular surveillance and phylogenetic traits of Babesia bigemina and Babesia bovis in cattle (Bos taurus) and water buffaloes (Bubalus bubalis) from Colombia

et al. 2018

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BackgroundBabesia bigemina and B. bovis are two economically important hemoparasites affecting both cattle and buffaloes involved in dairy and beef production. In Colombia, although some parasitological and serological studies suggest an endemicity of these pathogens in areas under 1000 m, little is known about its molecular prevalence in different host. The objective of this study was to estimate the prevalence and molecular traits of these parasites in cattle and buffaloes from two Colombian regions.MethodsBetween 2014 and 2016, a three-point longitudinal survey was designed in farms from Caribbean and Orinoquia regions to evaluate the molecular prevalence of B. bigemina and B. bovis using a nested PCR (n-PCR) targeting hypothetical protein (hyp) and rhoptry-associated protein (RAP-1) genes, respectively. A total of 1432 cattle, 152 buffalo and 1439 Rhipicephalus microplus samples were analyzed. Moreover, phylogenetic relationship of isolates was analyzed using the 18S rRNA gene.ResultsA molecular prevalence of 31.6% (24.2% for B. bigemina and 14.4% for B. bovis), 23.6% (6.5% for B. bigemina and 17.7% for B. bovis) and 4.3% (3.5% for B. bigemina and 1.0% for B. bovis) was observed in cattle, buffaloes and Rhipicephalus microplus, respectively. Higher values of infection were observed during the wet season and late wet season; nevertheless, other variables such as age, production type, sex, breed and babesiosis control were also significantly associated with infection. Prevalence analysis showed that B. bovis infection was higher in cattle that coexist with buffaloes, when compared to those which did not. For each species, phylogenetic analyses revealed a high genetic diversity of isolates without clusters related to the isolation source.ConclusionsTo our knowledge, this is the first longitudinal survey that evaluates through molecular methods, the infection of B. bigemina and B. bovis in two important livestock regions from Colombia. This study reveals that the prevalence of infection by Babesia spp., in cattle and buffaloes are modulated by seasonal variations, host factors and vector traits. Our results provide new insights on the epidemiological aspects of infection of Babesia spp., in cattle and buffaloes, which must be taken into consideration when babesiosis control programs are implemented in the study area.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-3091-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Molecular surveillance and phylogenetic traits of Babesia bigemina and Babesia bovis in cattle (Bos taurus) and water buffaloes (Bubalus bubalis) from Colombia

et al. 2018

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“…The following Argentinean B. bigemina strains were used: S1A, S2A, and M1A (all attenuated vaccine strains) and the pathogenic B38, S3P, and S2P strains. Details of the geographic origin of these strains and the respective multilocus genotypes have been described in a previous publication [16]. For the in silico analysis, the B. bigemina genomes of BOND, JG29, and the PR strains were used [12].…”

Section: B Bigemina Strains and Genomic Dna Isolationmentioning

confidence: 99%

Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

et al. 2020

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Background Thrombospondin-related anonymous protein (TRAP) has been described as a potential vaccine candidate for several diseases caused by apicomplexan parasites. However, this protein and members of this family have not yet been characterized in Babesia bigemina, one of the most prevalent species causing bovine babesiosis. Methods The 3186-bp Babesia bigemina TRAP-1 (BbiTRAP-1) gene was identified by a bioinformatics search using the B. bovis TRAP-1 sequence. Members of the TRAP and TRAP-related protein families (TRP) were identified in Babesia and Theileria through a search of the TSP-1 adhesive domain, which is the hallmark motif in both proteins. Structural modeling and phylogenetic analysis were performed with the identified TRAP proteins. A truncated recombinant BbiTRAP-1 that migrates at approximately 107 kDa and specific antisera were produced and used in Western blot analysis and indirect fluorescent antibody tests (IFAT). B-cell epitopes with neutralizing activity in BbiTRAP-1 were defined by enzyme-linked immunosorbent assays (ELISA) and invasion assays. Results Three members of the TRAP family of proteins were identified in B. bigemina (BbiTRAP-1 to -3). All are type 1 transmembrane proteins containing the von Willebrand factor A (vWFA), thrombospondin type 1 (TSP-1), and cytoplasmic C-terminus domains, as well as transmembrane regions. The BbiTRAP-1 predicted structure also contains a metal ion-dependent adhesion site for interaction with the host cell. The TRP family in Babesia and Theileria species contains the canonical TSP-1 domain but lacks the vWFA domain and together with TRAP define a novel gene superfamily. A variable number of tandem repeat units are present in BbiTRAP-1 and could be used for strain genotyping. Western blot and IFAT analysis confirmed the expression of BbiTRAP-1 by blood-stage parasites. Partial recognition by a panel of sera from B. bigemina-infected cattle in ELISAs using truncated BbiTRAP-1 suggests that this protein is not an immunodominant antigen. Additionally, bovine anti-recombinant BbiTRAP-1 antibodies were found to be capable of neutralizing merozoite invasion in vitro. Conclusions We have identified the TRAP and TRP gene families in several Babesia and Theileria species and characterized BbiTRAP-1 as a novel antigen of B. bigemina. The functional relevance and presence of neutralization-sensitive B-cell epitopes suggest that BbiTRAP-1 could be included in tests for future vaccine candidates against B. bigemina.

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“…The following Argentinean strains were used: S1A, S2A and M1A (all attenuated vaccine strains) and the pathogenic B38, S3P and S2P strains. Details of the geographic origin of these strains and the respective multilocus genotypes were already described (14). For in silico analysis, the B. bigemina genomes of BOND, JG29 and PR strains were used (10).…”

Section: B Bigemina Strains and Genomic Dna Isolationmentioning

confidence: 99%

Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

Montenegro

Paoletta

Ortiz

et al. 2020

Preprint

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Background The Thrombospondin-Related Anonymous Protein (TRAP) has been described as a potential vaccine candidate in several apicomplexan parasites. However, this protein and members of this family have not been characterized yet in Babesia bigemina, one of the most prevalent species causing bovine babesiosis.Methods The Babesia bigemina TRAP-1 (BbiTRAP-1) gene was identified by a bioinformatics search using the B. bovis TRAP-1 sequence. Members of the TRAP and TRAP-related protein family (TRP) families were identified in Babesia and Theileria through the search of the TSP-1 adhesive domain, the hallmark motif of TRAP proteins. Structural modelling and phylogenetic analysis were performed with the identified TRAP proteins. A truncated recombinant BbiTRAP-1 and specific antisera were produced, and used in Western blot analysis and indirect fluorescent antibody test (IFAT). B-cell epitopes with neutralizing activity in BbiTRAP-1 were defined by ELISA and invasion assays.Results TRAP family has 3 members in B. bigemina (BbiTRAP-1-3). All are type 1 transmembrane proteins containing the von Willebrand factor A (vWFA), thrombospondin type 1 (TSP-1) and cytoplasmic C-terminus domains along with transmembrane regions. The BbiTRAP-1 predicted structure also contains a metal ion-dependent adhesion site (MIDAS) for interaction with the host cell. The TRP family in Babesia and Theileria species contains the canonical TSP-1 domain but lack the vWFA domain,and together with TRAP define a novel gene superfamily. A variable number of tandem repeat units is present in BbiTRAP-1 and could be used for strain genotyping. Western blot and IFAT analysis confirmed expression of BbiTRAP-1 by blood stage parasites. Partial recognition by a panel of sera from B. bigemina infected cattle in ELISA using truncated BbiTRAP-1 suggests that this protein is not an immunodominant antigen. Additionally, bovine anti- recombinant BbiTRAP-1 antibodies were capable of neutralize merozoite invasion in vitro. Conclusions We identified the TRAP and TRP gene families in several Babesia and Theileria species, and characterized BbiTRAP-1 as a novel antigen of B. bigemina. The functional relevance and presence of neutralization-sensitive B-cell epitopes suggest that BbiTRAP-1 could be included in future vaccine candidates against B. bigemina.

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Multi-locus typing scheme for Babesia bovis and Babesia bigemina reveals high levels of genetic variability in strains from Northern Argentina

Cited by 13 publications

References 44 publications

Molecular surveillance and phylogenetic traits of Babesia bigemina and Babesia bovis in cattle (Bos taurus) and water buffaloes (Bubalus bubalis) from Colombia

Molecular surveillance and phylogenetic traits of Babesia bigemina and Babesia bovis in cattle (Bos taurus) and water buffaloes (Bubalus bubalis) from Colombia

Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

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