Accurate automated quantitative imaging of tortoise erythrocytes using the NIS image analysis system

The standard method for assessing blood cell characteristics using an ocular micrometer is time-consuming and limited. We used the Nikon NIS Elements imaging software and May-Grünwald-Giemsa staining to determine whether automated image analysis is suitable for rapid and accurate quantitative morphometry of erythrocytes. Blood was collected during four seasons from 126 geometric tortoises and the blood smears were evaluated for cell (C) and nuclear (N) characteristics of the erythrocytes. We measured area, length (L), width (W), perimeter, elongation and pixelation intensity, and calculated L/W and N/C areas. Erythrocyte size differed among cohorts; females, the larger sex, had smaller erythrocytes than either males or juveniles. Males had more elongated erythrocytes than females and erythrocytes of adults were more elongated than those of juveniles. Erythrocyte size and shape influence the efficiency of gas exchange owing to surface area to volume ratios, which are greater for small, elongated cells than for large, round cells. The high N/C ratio and low pixelation intensities of males and juveniles indicate that they may have had more immature erythrocytes in their circulation than females. The use of pixelation intensity to indicate the presence of immature erythrocytes was validated by seasonal differences that corresponded to the biology of the tortoises. Pixelation intensity was lowest in winter. We found that automated image analysis is a rapid and reliable method for determining cell size and shape, and it offers the potential for distinguishing among developmental stages that differ in staining intensity. The method should be useful for rapid health assessments, particularly of threatened species, and for comparative studies among different vertebrates.Web of Scienc

Morphological characterization of the blood cells in the endangered Sicilian endemic pond turtle,Emys trinacris(Testudines: Emydidae)

In this study, measurements of morphological parameters, sizes and frequencies of peripheral blood cells (erythrocytes, leukocytes, thrombocytes) on blood smear preparation devices stained with May-Grünwald stain were evaluated for both sexes in 20 Emys trinacris (Testudines: Emydidae) specimens. Erythrocytes were higher in male than in female specimens. The leukocyte of E. trinacris contains eosinophil, basophil, monocyte, heterophil and lymphocyte. The eosinophil was higher in males than in females whereas lymphocytes were higher in females than in males. The erythrocyte morphological parameters (EL [erythrocyte length], EW [erythrocyte width], L/W [length/width], ES [erythrocyte size]) were compared with the same data from Emys orbicularis s.l, and from species belonging to other chelonian genera. The erythrocyte size did not vary within the studied Palearctic Emys taxa, whereas it proved to differ from that observed in other chelonians

(Ophidia: Colubridae), collected from North-Western Turkey

In this investigation, 11 Smooth snakes (Coronella austriaca), and 8 Aesculapian snakes (Zamenis longissimus) were collected between 1998 and 2007 from northwestern Turkey and were examined for the first time helminthologically. It was recorded that Coronella austriaca harbored 1 species of Nematoda (Kalicephalus sp.) and one unidentified nematode cyst; Zamenis longissimus, 4 species of Nematoda (Rhabdias fuscovenosa, Oxysomatium brevicaudatum, Kalicephalus sp. and Ophidascaris sp.) and two unidentified cysts. All helminths samples represented new host records for Coronella austriaca and Zamenis longissimus in Turkey. It was also recorded that Turkey is a new locality for Kalicephalus sp. and Ophidascaris sp

worm, Anguis fragilis Linnaeus 1758 (Squamata: Anguidae), from Turkey

In this investigation, seventeen Turkish worm lizards, Blanus strauchi, and eighteen slow worms, Anguis fragilis, collected from Turkey, were examined for helminths. Blanus strauchi harbored 2 species of Nematoda: Pharyngodon spinicauda and Aplectana sp. (larvae); Anguis fragilis harbored 5 species of Nematoda:, Rhabdias bufonis, Entomelas entomelas, Oswaldocruzia filiformis, Cosmocerca ornata and Oxysomatium brevicaudatum. Blanus strauchi represents a new host record for Pharyngodon spinicauda and Aplectana sp. Anguis fragilis, represents a new host record for Cosmocerca ornata. Turkey is a new locality record for Pharyngodon spinicauda

(Ophidia: Colubridae), collected from North-Western Turkey

In this investigation, 11 Smooth snakes (Coronella austriaca), and 8 Aesculapian snakes (Zamenis longissimus) were collected between 1998 and 2007 from northwestern Turkey and were examined for the first time helminthologically. It was recorded that Coronella austriaca harbored 1 species of Nematoda (Kalicephalus sp.) and one unidentified nematode cyst; Zamenis longissimus, 4 species of Nematoda (Rhabdias fuscovenosa, Oxysomatium brevicaudatum, Kalicephalus sp. and Ophidascaris sp.) and two unidentified cysts. All helminths samples represented new host records for Coronella austriaca and Zamenis longissimus in Turkey. It was also recorded that Turkey is a new locality for Kalicephalus sp. and Ophidascaris sp

(Anura: Ranidae) collected from Northwest of Turkey

A total of 33 agile frogs (Rana dalmatina) were collected from 7 localities in Edirne, Bursa and AdapazarA +/- Provinces (Northwest of Turkey), between 1987-2007 and examined for the first time for helminths. R. dalmatina harbored one species of Monogenea (Polystoma sp.), 3 species of Digenea (Diplodiscus subclavatus, Pleurogenoides medians, and Pleurogenes claviger), 4 species of Nematoda (Rhabdias bufonis, Oswaldocruzia filiformis, Cosmocerca ornata, and Oxysomatium brevicaudatum) and one species of Acanthocephala (Acanthocephalus ranae). All helminths represent new host records for Rana dalmatina in Turkey