RESEARCH PAPER
Comparison of the detection efficiency of haemoparasite DNA in blood and faecal samples – the way to eco-epidemiological studies
 
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University of Warsaw, Poland
CORRESPONDING AUTHOR
Anna Bajer   

University of Warsaw, Miecznikowa 1, 02-096, Warszawa, Poland
 
KEYWORDS
TOPICS
ABSTRACT
Introduction and objective:
It is easier and non-invasive to obtain faecal samples compared with blood samples. Molecular techniques may enable detection of parasites even in tiny amounts of blood-containing faeces. We aimed to compare the sensitivity of detection of three Babesia species and Hepatozoon canis in blood and faecal samples, including samples derived from naturally infected hosts.

Material and methods:
Three groups were involved: 1) Nine BALB/c mice infected with Babesia microti sampled during acute (n=3), post-acute (n=3) and chronic phases of infection (n=3); 2) Eight dogs with symptoms of babesiosis; 3) Six red foxes infected with B. vulpes, one fox infected with B. canis, four foxes infected with H. canis. Genomic DNA was extracted from blood and faeces by use of commercial kits and amplified with genus-specific primers in one-step or nested PCR reactions. Selected PCR products were sequenced.

Results:
No positive results for faecal samples were obtained from H. canis-positive foxes in contrast to Babesia spp. infections. Positive results from PCRs were obtained for all BALB/c mice (100%), five dogs (62.5%) and four of seven foxes (57.1%). Successful sequencing was obtained for six selected murine samples (B. microti), four canine samples (B. canis) and for one fox sample (B. vulpes). The success of B. microti detection in murine faecal samples from acute, post-acute and chronic phases was identical (100%).

Conclusions:
Detectability of Babesia spp. infections was lower in naturally infected dogs and foxes, compared to experimentally infected mice. Detection of DNA in faecal samples can be useful in the detection of Babesia infection in populations from which blood samples are hard to obtain, but due regard must be given to the possibility that prevalence of infection may be severely underestimated.

ACKNOWLEDGEMENTS
The authors express their thanks to the owners of the dogs for their kind cooperation. Professor Jerzy M. Behnke at the University of Nottingham, UK, undertook a critical review of the manuscript. The study was partially supported by the National Science Centre (NCN), grant Sonata Bis No. 2014/14/E/NZ7/00153, who had no role in the design of the study and collection of samples, analysis and interpretation of data and, or in the writing of the manuscript.
 
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