Babesia Canis Canis, Babesia Canis Vogeli, Babesia Canis Rossi: Differentiation of the Three Subspecies By A Restriction Fragment Length Polymorphism Analysis On Amplified Small Subunit Ribosomal Rna Genes

Abstract

The parasites Babesia canis and Babesia gibsoni (phylum Apicomplexa) are responsible for canine babesiosis throughout the world. Babesia canis was previously described as a group of three biologically different subspecies, namely B. canis canis, B. canis vogeli, and B. canis rossi. We report partial sequences of small subunit ribosomal RNA gene (ssu-rDNA) of each subspecies amplified in vitro with primers derived from a semi-conserved region of the ssu-rDNA genes in other Babesia species. The polymerase chain reaction combined with a restriction fragment length polymorphism analysis, using HinfI and TaqI restriction enzymes, confirmed the separation of B. canis into three subspecies. These sequences were compared with previously published sequences of other Babesia species. A phylogenetic approach showed that the three subspecies of B. canis belong to the clade of Babesia species sensu stricto where B. canis canis clusters with B. canis rossi whereas B. canis vogeli might form a monophyletic group with the cluster B. divergens and B. odocoilei. Our results show that the three subspecies of B. canis can readily be differentiated at the molecular level and suggest that they might be considered as true species.

Retrospective Analysis of Canine Vector-borne Diseases (CVBD) in Germany with Emphasis on the Endemicity and Risk Factors of Leishmaniosis

“…(15,155 samples) and E. canis (570 samples), DNA was extracted from whole blood samples according to Dyachenko et al (2012), and a conventional PCR was performed according to Carret et al (1999), targeting the SSU rDNA gene (Babesia spp.) and to McBride et al (1996), targeting the 16S ribosomal RNA gene (E. canis).…”

Diagnosis of Imported Canine Filarial Infections in Germany 2008 – 2010

“…DNA was performed by means of a conventional PCR according to Carret et al (1999). No further differentiation was performed on samples with positive detection of Babesia DNA.…”

Detection and molecular characterization of piroplasms species from naturally infected dogs in southeast Brazil

“…To define the species of Babesia, the amplification products were digested with the enzymes Hinf I (Biolabs®, New England) and Taq I (Biolabs®, New England) for 3 hours (CARRET et al, 1999) and then analyzed by 2% agarose gel electrophoresis.…”

“…The program consisted in an initial denaturation step at 94 °C for 5 minutes, followed by 30 denaturation cycles at 94 °C for 1 minute, annealing at 55 °C for 1 minute and extension at 72 °C for 1 minute. A final extension step at 72 °C for 5 minutes was used (CARRET et al, 1999).…”