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Background  Classification of  Babesia  parasites has traditionally relied on morphological differentiation based on piroplasm size and shape. Molecular typing has subsequently revealed a more complex taxonomy for these piroplasms than previously thought. To evaluate the factors that influence the morphology of  Babesia  species upon microscopic examination and hence, their taxonomic classification, we performed detailed characterizations of piroplasms from archival and prospective collections of cytological samples of dogs with piroplasmosis before and after death. Merozoite morphology and time of parasite disappearance following imidocarb dipropionate was also investigated.    Methods  The study was divided into a (i) review of archived cytological slides from confirmed cases of canine piroplasmosis, and (ii) a prospective study of smears and tissue imprints from 15 recently necropsied dogs. The latter group could be further sub-divided into a non-treated group and an imidocarb dipropionate-treated group. Exact times of treatment before death were reviewed. Additional blood smears prepared from the live dogs and taken before therapy were also evaluated in the latter group. Parasite burden per each slide was determined in both studies. The shape and size of merozoites were described from blood smears taken while the dogs were alive and from different organs during necropsy. The results of all measurements were statistically analyzed.    Results  The morphology and size of merozoites from live dogs corresponded to that of previously described ‘large’  Babesia . The morphology and size of merozoites were significantly different ( P  < 0.001) in postmortem samples, however, and more consistent in shape and size with piroplasm cells previously referred to as ‘small’  Babesia . PCR and sequencing confirmed  B. canis  as the causative agent of disease in all investigated dogs, including in postmortem negative tissue imprints from dogs treated at least 24 h before death.    Conclusions  Changes in the morphology of ‘large’  B. canis  to ‘small’-like  Babesia  observed by light microscopy appear to represent a common postmortem change. Classification of  Babesia  parasites into ‘large’ and ‘small’  Babesia  using only microscopy of postmortem slides should be treated with caution. PCR-based methodologies for detection and molecular typing of  Babesia  spp. may prove valuable for investigating suspected cases of babesiosis following necropsy.    Electronic supplementary material  The online version of this article (10.1186/s13071-017-2412-1) contains supplementary material, which is available to authorized users.

parasites and vectors

Microscopic and molecular analysis of Babesia canis in archived and diagnostic specimens reveal the impact of anti-parasitic treatment and postmortem changes on pathogen detection