Typification of virulent and low virulence Babesia bigemina clones by 18S rRNA and rap-1c

The population structure of original Babesia bigemina isolates and reference strains with a defined phenotypic profile was assessed using 18S rRNA and rap-1c genes. Two reference strains, BbiS2P-c (virulent) and BbiS1A-c (low virulence), were biologically cloned in vitro. The virulence profile of the strains and clones was assessed in vivo. One fully virulent and one low-virulence clone were mixed in identical proportions to evaluate their growth efficiency in vitro. Each clone was differentiated by two microsatellites and the gene gp45. The 18S rRNA and rap-1c genes sequences from B. bigemina biological clones and their parental strains, multiplied exclusively in vivo or in vitro, were compared with strain JG-29. The virulence of clones derived from the BbiS2P-c strain was variable. Virulent clone Bbi9P1 grew more efficiently in vitro than did the low-virulence clone Bbi2A1. The haplotypes generated by the nucleotide polymorphism, localized in the V4 region of the 18S rRNA, allowed the identification of three genotypes. The rap-1c haplotypes allowed defining four genotypes. Parental and original strains were defined by multiple haplotypes identified in both genes. The rap-1c gene, analyzed by high-resolution melting (HRM), allowed discrimination between two genotypes according to their phenotype, and both were different from JG-29. B. bigemina biological clones made it possible to define the population structure of isolates and strains. The polymorphic regions of the 18S rRNA and rap-1c genes allowed the identification of different subpopulations within original B. bigemina isolates by the definition of several haplotypes and the differentiation of fully virulent from low virulence clones.

Bibliographic Details

Main Authors:	Thompson, Carolina Soledad, Baravalle, María Eugenia, Valentini, Beatriz Susana, Mangold, Atilio Jose, Torioni, Susana Marta, Ruybal, Paula, Farber, Marisa Diana, Echaide, Ignacio Eduardo
Format:	info:ar-repo/semantics/artículo biblioteca
Language:	eng
Published:	2014-06
Subjects:	Babesia bigemina, ARN, Virulencia, Clones, Genética, RNA, Virulence, Genetics, 18S rRNA,
Online Access:	https://www.sciencedirect.com/science/article/pii/S0014489414000599 http://hdl.handle.net/20.500.12123/3031 https://doi.org/10.1016/j.exppara.2014.03.016
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