Cytological and molecular identification of haemogregarina stepanowi in blood samples of the european pond turtle (emys orbicularis) from quarantine at belgrade zoo

Abstract

Blood smears stained with Diff Quick are the initial tool for cytological diagnosis of Haemogregarina spp. However, the development of sensitive and specifi c molecular methods enabled the detection and identifi cation of parasites in the sample and to clarify the evolutionary relationships of adeleorinid parasites within the Apicomplexa. The current study was attempted in order to perform cytological investigation and molecular identifi cation of the hemoparasites in thirty European pond turtles (Emys orbicularis) from the quarantine section at Belgrade Zoo, which have been found in poor health condition with massive skin hemorrhages, based on intraerythrocytic parasitic forms on hematological smears and 18S rDNA sequence, respectively. Different life cycle stages of the Haemogregarina sp. were noticed within the erythrocytes in the peripheral blood. Biochemical analysis indicated lower values of AST and iron in most of the infected turtles while hematological analysis showed a changed hematocrit value, a decrease in the number of red blood cells and low hemoglobin levels. Amplifi cations of the 18S rDNA sequence of Haemogregarina were detected in 30/30 (100%) turtles with clinical symptoms. The identity of PCR products was confi rmed by direct DNA sequencing. Future research concerning H. stepanowi in Serbia should be applied to its defi nitive host-the leech.