Evaluation of doxycycline sensitivity in vitro against Babesia gibsoni by Real Time-PCR

A Matsuu,Mh Talukder,Y Hikasa

doi:10.3329/jbau.v11i1.18219

A Matsuu, Mh Talukder + Show 1 more

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https://doi.org/10.3329/jbau.v11i1.18219

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Abstract

Doxycycline sensitivity against in vitro cultured Babesia gibsoni was evaluated by real-time PCR and parasitemia. The culture of B. gibsoni was successfully continued and mean parasitemia was 4.63% when in vitro drug sensitivity test was started. The drug sensitivity test by the real-time PCR calculated the gene copy number from cultured sample. Even if the complete growth inhibition was at certain concentration, the genomic DNA might remain in existence. The results revealed that the doses of doxycycline 8?M, 16 ?M, 32 ?M and 64 ?M of both real-time PCR and parasitemia inhibited the growth of B. gibsoni in a dose-dependent manner at 96 h and 144 h. We determined the IC50 of doxycycline 17.9 ?M at 96 h and 13.8 ?M at 144 h after treatment using real-time PCR. Morphological observation revealed that the number of parasites decreased per erythrocyte and line shaped parasites increased in erythrocytes with the increased concentration of doxycycline. Doxycycline effectively inhibits the growth of B. gibsoni in vitro. Therefore, doxycycline can be used to treat B. gibsoni infection. Further studies are important to know the doxycycline resistance in B. gibsoni. DOI: http://dx.doi.org/10.3329/jbau.v11i1.18219 J. Bangladesh Agril. Univ. 11(1): 97-102, 2013

Highlights

B. gibsoni is a tick-borne intra-erythrocytic protozoan parasite
There is Evaluation of doxycycline sensitivity in vitro against Babesia gibsoni no precise report emphasizing the efficacy of doxycycline sensitivity alone in vitro against B. gibsoni by Real Time-PCR (RT-PCR)
The growth inhibitory effect of different concentrations of doxycycline against B. gibsoni tested in a continued culture and evaluated by RT-PCR at different time points

Summary

Introduction

B. gibsoni is a tick-borne intra-erythrocytic protozoan parasite. It is one of the most common causal agents of severe canine haemolytic anemia. To evaluate the effect of potential drugs against this pathogen, an in vitro susceptibility test would be important by Real Time-PCR (RT-PCR) but limited information is available. B. gibsoni infected dogs either clinical patient or experimental are required for this method.

Results

Conclusion