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.

Cytochrome c oxydase I phylogenetic analysis of Haemogregarina parasites (Apicomplexa, Coccidia, Eucoccidiorida, Haemogregarinidae) confirms the presence of three distinct species within the freshwater turtles of Tunisia

“…Based on morphological and molecular analyses, a low diversity of Haemogregarina species, accounting for less than 20 species [31,32,[48][49][50][51][52][53][54], successfully infect freshwater turtles. With the exception of Attia El Hili et al [32] who reported from 18S rRNA gene sequences the presence of three Haemogregarina species within the same individuals, all other reports that focused on both morphological and genetic data of Haemogregarina revealed the presence of single or low divergent 18S variants per individual, suggesting the occurrence of a single parasite species in sympatric hosts [31,[48][49][50][51][52][53][54]. Following PCR assays that were conducted on haemosporidian parasites, Bernotienė et al [47] showed several cases of mixed infections indicating that single PCR assays could underestimate the true parasitic diversity.…”

The good, the bad and the ugly: Emys trinacris, Placobdella costata and Haemogregarina stepanowi in Sicily (Testudines, Annelida and Apicomplexa)

“…A partial sequence of the nuclear 18S rRNA gene was amplified through the specific apicomplexan primers (EF: 5'-GAAACTGCGAATGGCTCATT-3' and ER: 5'-TTG-CGCCTACTAGGCATTC-3') designed by Kvičerová et al (2008). The target fragment is considered sufficiently variable and informative at both the generic and specific levels in Apicomplexa (Barta et al 2012, Dvořáková et al 2014, Özvegy et al 2015.…”

Systemic Granulomatous Pathology in Two Captive Alligator Mississippiensis

“…Many of them have been reported producing lesions of the skin or of the lungs, and fi nally being incriminated in developing septicemia and lead to the death of the animal. The wide majority of the animals suffered from different types of stress, they were of different ages and have been captive in farms, circuses or zoos [2,12,27]. In both alligators, wood, stone and glass fragments were found in the stomach, in the absence of food, causing erosions.…”