Chromosome-level genome assembly of Babesia caballi reveals diversity of multigene families among Babesia species

Ochi, Akihiro; Kidaka, Taishi; Hakimi, Hassan; Asada, Masahito; Yamagishi, Junya

doi:10.1186/s12864-023-09540-w

Cited by 3 publications

(1 citation statement)

References 53 publications

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“…Xinjiang , T. equi , P. falciparum , T. gondii and B. caballi ). Finally, it is important to note that B. caballi was phylogenetically closer to B. bigemina and B. ovata than to B. bovis [ 47 ].…”

Section: Discussionmentioning

confidence: 99%

Comparative Transcriptome Analysis of Babesia bigemina Attenuated Vaccine and Virulent Strains of Mexican Origin

Santamaria,

Estrada,

López

et al. 2024

Vaccines

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Bovine babesiosis, caused by the protozoan Babesia bigemina, is one of the most important hemoparasite diseases of cattle in Mexico and the world. An attenuated B. bigemina strain maintained under in vitro culture conditions has been used as a live attenuated vaccine; however, the biological mechanisms involved in attenuation are unknown. The objective of this study was to identify, through a comparative transcriptomics approach, the components of the B. bigemina virulent parasites that are differentially expressed in vivo, as opposed to those expressed by B. bigemina attenuated vaccine parasites when inoculated into naïve cattle. The biological material under study was obtained by inoculating spleen-intact cattle with infected erythrocytes containing either the attenuated strain or a virulent field strain. After RNA extraction, transcriptomic analysis (RNA-seq) was performed, followed by bioinformatic Differential Expression (DE) analysis and Gene Ontology (GO) term enrichment. The high-throughput sequencing results obtained by analyzing three biological replicates for each parasite strain ranged from 9,504,000 to 9,656,000, and 13,400,000 to 15,750,000 reads for the B. bigemina attenuated and virulent strains, respectively. At least 519 differentially expressed genes were identified in the analyzed strains. In addition, GO analysis revealed both similarities and differences across the three categories: cellular components, biological processes, and molecular functions. The attenuated strain of B. bigemina derived from in vitro culture presents global transcriptomic changes when compared to the virulent strain. Moreover, the obtained data provide insights into the potential molecular mechanisms associated with the attenuation or pathogenicity of each analyzed strain, offering molecular markers that might be associated with virulence or potential vaccine candidates.

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Section: Discussionmentioning

confidence: 99%

Comparative Transcriptome Analysis of Babesia bigemina Attenuated Vaccine and Virulent Strains of Mexican Origin

Santamaria,

Estrada,

López

et al. 2024

Vaccines

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ves1αgenes expression is the major determinant ofBabesia bovis-infected erythrocytes cytoadhesion to endothelial cells

Hakimi,

Yamagishi,

Sakaguchi

et al. 2024

Preprint

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Babesia boviscauses the most pathogenic form of babesiosis in cattle, resulting in high mortality in naive adults. This parasite invades red blood cells (RBCs) within the bovine hosts where they multiply and produce clinical disease.Babesia bovisexports numerous proteins into invaded RBCs changing its properties. Thus, the infected RBCs (iRBCs) are capable to cytoadhere in the microvasculature of internal organs and brain, leading to respiratory distress, neurologic signs, and mortality. Variant Erythrocyte Surface Antigen 1 (VESA1) is one of those exported proteins byB. boviswhich represents a major virulence factor due to its central role in immune evasion by antigenic variation and intravascular parasite sequestration. VESA1 is a heterodimer protein encoded byves1αandves1βmultigene family and localized on the ridges, the focal point for cytoadhesion. To gain further insights into the molecular mechanisms of cytoadhesion ofB. bovis, we panned the parasites with bovine brain microvasculature endothelial cells, which resulted in obtaining several clones with different cytoadherence abilities. The transcriptome analysis of 2 high and 2 low cytoadherent clones revealed thatves1αsequences were diversified, likely resulting from genomic recombination. On the other hand,ves1βsequences were almost identical among these 4 clones. Insertion and expression ofves1αof a clone with high binding intoef-1αlocus of a low binging clone increased cytoadherence confirming the role ofves1αsuggested by our transcriptome data. Whole genome sequencing of cytoadherent clones revealed active locus of ves1 on chromosome 2. These results suggest that VESA1a proteins encoded byves1αgenes determine the cytoadherence specificity and/or cytoadherence strength ofB. bovisand they are in the active site for recombination.

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In vitro evaluation of the automated hematology analyzer XN-31 for rapid diagnosis of equine piroplasmosis

Ochi,

Toya,

Sengoku

et al. 2024

Microbiol Spectr

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Equine piroplasmosis (EP) is a protozoal disease affecting equids, caused by Theileria equi and Babesia caballi . EP is conventionally diagnosed using microscopic, molecular, and/or serological methods, which are time-consuming. Consequently, there is a need for faster testing methods. In this study, we evaluated the application of the Sysmex XN-31 automated hematology analyzer, originally a rapid test for detecting malaria in humans, for the diagnosis of EP. The cultured parasites were measured using the XN-31 that had been customized for horse blood samples (XN-31m). The following parameters were evaluated: limit of detection (LoD), limit of quantification (LoQ), linearity, carryover, precision, and correlation with microscopic examination. The XN-31m detected infected red blood cells (RBCs) in approximately 1 minute. The LoD and LoQ for B. caballi were 4.54 infected RBCs/μL and 14.10 infected RBCs/μL, while those for T. equi were 5.80 infected RBCs/μL and 11.44 infected RBCs/μL, respectively. Linearity showed excellent correlation ( R 2 > 0.99), and carryover never exceeded 0.5%. The coefficient of variation was under 5%. The correlation between the results obtained using XN-31m and microscopic examination was high ( R 2 > 0.98). In conclusion, the XN-31 analyzer detected B. caballi and T. equi parasites in approximately 1 minute with high sensitivity. The results indicate the potential of the XN-31 analyzer as a fast and user-friendly diagnostic method for EP. IMPORTANCE In this study, we demonstrated that the automated hematology analyzer, XN-31, can detect red blood cells infected with Babesia caballi and Theileria equi in about 1 minute. We evaluated the diagnostic performance of the XN-31 analyzer for equine piroplasmosis, providing evidence of its potential as a diagnostic tool for this disease.

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Chromosome-level genome assembly of Babesia caballi reveals diversity of multigene families among Babesia species

Cited by 3 publications

References 53 publications

Comparative Transcriptome Analysis of Babesia bigemina Attenuated Vaccine and Virulent Strains of Mexican Origin

Comparative Transcriptome Analysis of Babesia bigemina Attenuated Vaccine and Virulent Strains of Mexican Origin

ves1αgenes expression is the major determinant ofBabesia bovis-infected erythrocytes cytoadhesion to endothelial cells

In vitro evaluation of the automated hematology analyzer XN-31 for rapid diagnosis of equine piroplasmosis

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