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Background   Hepatozoon canis  is a protozoan tick-borne pathogen of dogs and wild canids.  Hepatozoon  spp. have been reported to infect foxes in different continents and recent studies have mostly used the polymerase chain reaction (PCR) for the detection and characterization of the infecting species. Surveying red foxes ( Vulpes vulpes ) may contribute to better understanding the epidemiology of canine vector-borne diseases, including hepatozoonosis caused by  H. canis  in domestic dogs. The present study investigated the prevalence of  Hepatozoon  spp. by means of histopathology and molecular analysis of different tissues in red foxes from different parts of Portugal.    Methods  Blood and tissues including bone marrow, heart, hind leg muscle, jejunum, kidney, liver, lung, popliteal or axillary lymph nodes, spleen and/or tongue were collected from 91 red foxes from eight districts in northern, central and southern Portugal. Tissues were formalin-fixed, paraffin-embedded, cut and stained with hematoxylin and eosin. Polymerase chain reaction (PCR) amplified a ~650 bp fragment of the 18S rRNA gene of  Hepatozoon  spp. and the DNA products were sequenced.    Results   Hepatozoon canis  was detected in 68 out of 90 foxes (75.6%) from all the sampled areas by PCR and sequencing. Histopathology revealed  H. canis  meronts similar in shape to those found in dogs in the bone marrow of 11 (23.4%) and in the spleen of two (4.3%) out of 47 foxes ( p  = 0.007). All the 11 foxes found positive by histopathology were also positive by PCR of bone marrow and/or blood. Positivity by PCR (83.0%) was significantly higher ( p  < 0.001) than by histopathological examination (23.4%) in paired bone marrow samples from the same 47 foxes. Sequences of the 18S rRNA gene of  H. canis  were 98–99% identical to those in GenBank.    Conclusions   Hepatozoon canis  was found to be highly prevalent in red fox populations from northern, central and southern Portugal. Detection of the parasite by histopathology was significantly less sensitive than by PCR. Red foxes are a presumptive reservoir of  H. canis  infection for domestic dogs.

Molecular and histopathological detection of Hepatozoon canis in red foxes (Vulpes vulpes) from Portugal