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Differentiation between Israeli B. bovis vaccine strain and field isolates
T Molad1, L Fleiderovitz1, B Leibovitz1
1Division of Parasitology, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
Veterinary Parasitology
|February 1, 2015
Summary
This study developed a PCR-RFLP method to differentiate Israeli Babesia bovis vaccine strains from field isolates. This technique identifies unique genetic markers in the rap-1 gene, aiding in molecular epidemiology.
Area of Science:
- Veterinary Parasitology
- Molecular Biology
- Genetics
Background:
- Babesia bovis causes significant economic losses in cattle globally.
- Distinguishing between vaccine strains and field isolates of B. bovis is crucial for effective disease control and epidemiological studies.
- Previous methods lacked the specificity to reliably differentiate between vaccine and field strains in Israel.
Purpose of the Study:
- To develop a molecular method for distinguishing the Israeli Babesia bovis vaccine strain from field isolates.
- To analyze genetic variations in the rhoptry-associated protein-1 (rap-1) gene between vaccine and field strains.
- To provide insights into the molecular epidemiology of B. bovis in Israel.
Main Methods:
- Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) assay.
- Targeted amplification and analysis of the rhoptry-associated protein-1 (rap-1) gene.
- Sequence alignment of deduced amino acid sequences using ClustalW.
Main Results:
- Identified unique EcoRI and HaeIII restriction site variations in the rap-1 gene of the Israeli vaccine strain.
- Achieved 97.5%–100% amino acid sequence identity among Israeli B. bovis strains and field isolates.
- Observed approximately 90% amino acid identity between Israeli strains and B. bovis from other countries (Argentina, Mexico, Brazil, USA).
Conclusions:
- The developed PCR-RFLP assay effectively distinguishes between Israeli B. bovis vaccine and field isolates, even in mixed populations.
- This method offers a valuable tool for monitoring vaccine efficacy and understanding B. bovis molecular epidemiology in Israel.
- Genetic variations in the rap-1 gene provide a basis for differentiating B. bovis strains.
