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Background  Parasites of the family Spirorchiidae cause disease and mortality in marine and freshwater turtles; two species,  Hapalotrema mistroides  and  Neospirorchis  sp., are reported in the resident population of loggerhead turtles of the Mediterranean Sea, with the first being the most widespread. In vivo diagnosis of spirorchidiasis can represent a challenge in guaranteeing prompt control and treatment of the disease and is currently limited to copromicroscopy.  The aim of this study was the development of a real time PCR assay with TaqMan probe for the detection of  H. mistroides  infection in the blood of live loggerhead turtles,  Caretta caretta,  hospitalized in rehabilitation centres. Its potential use for in vivo diagnosis is explored.    Results  The developed real time PCR successfully detected  H. mistroides  DNA from both positive controls and experimental blood samples of live loggerhead sea turtles, showing good specificity, sensitivity and good reaction efficiency. Two out of three turtles which had demonstrated positivity at copromicroscopy also tested positive to this blood assay; DNA of  H .  mistroides  was detected within the blood of one sea turtle, which tested negative for copromicroscopy.    Conclusions  This study describes a specific and rapid molecular assay to detect  H. mistroides  infection from live sea turtles and highlights for the first time the presence of DNA of this species in turtle blood samples. Since this assay is able to detect low amounts of the parasitic free DNA in blood samples, its application could be helpful for in vivo diagnosis of  H. mistroides  infection as well as for epidemiological purposes.

bmc veterinary research

A pilot study on molecular diagnosis of Hapalotrema mistroides (Digenea: Spirorchiidae) infection in blood samples of live loggerhead turtles Caretta caretta