The Russian Tortoise 
Agrionemys horsfieldii ( Testudo horsfieldii)  


Knotková Z., J. Doubek, Z. Knotek, P. Hájková: Blood Cell Morphology and Plasma Biochemistry in Russian Tortoises (Agrionemys horsfieldi).
Acta Vet. Brno 2002, 71: 191-198.
Morphological characteristics of peripheral blood cells and plasma biochemistry profile were examined in twenty clinically healthy Russian tortoises (Agrionemys horsfieldi). The blood samples were collected from dorsal coccygeal vein. The Pappenheim staining method was used as the basal method for the classification of the blood cells. For enriching staining panel the commercial kits Rapidiffâ , Lysocolorâ , Lymphocolorâ , Monocolorâ , Granulocolorâ and Neutrocolorâ were used. Ten different types of blood cells were determined: erythrocytes, thrombocytes, lymphocytes, monocytes, type-I cells (heterophils), type-II cells (eosinophils), type-III cells (azurophils), type-IV cells (basophils), type-Ia cells (toxic heterophils) and type-V cells (polychromatophil erythrocytes). Stained with Lysocolorâ , the positive reaction emerged in type-IV cells and less in type-III cells. Granulocolorâ stained positively type-I, Ia and II cells. General purpose stain Rapidiffâ was less vivid than the classical Pappenheim method. The differential count of leukocytes in the peripheral blood of clinically healthy tortoises was determined as follows: 0.372 ± 0.089 heterophils, 0.048 ± 0.011 eosinophils, 0.051 ± 0.020 azurophils, 0.050 ± 0.009 basophils, 0.467 ± 0.125 lymphocytes and 0.012 ± 0.006 monocytes. Plasma concentrations of total protein, glucose, uric acid, cholesterol, natrium, potassium, calcium, phosphorus and activities of ALP, ALT, AST were: 45.1± 6.8 g/l, 11.4 ± 1.3 mmol/l, 95.0 ± 20.1 m mol/l, 4.0 ± 1.7 mmol/l, 143.0 ± 6.5 mmol/l, 3.8 ± 0.9 mmol/l, 2.5 ± 0.9 mmol/l, 1.4 ± 0.4 mmol/l, and 4.5 ± 2.0 m kat/l, 0.8 ± 0.4 m kat/l, 1.1 ± 0.3 m kat/l, respectively. The differential count of leukocytes and biochemical values may be used as a standard profile for healthy Russian tortoises kept in captivity. Haematology, Reptilia, Chelonia, Leukocytes

Address for correspondence:
MVDr. Z. Knotková, CSc.
Department of Physiology, Faculty of Vet.
Medicine University of Veterinary and Pharmaceutical Sciences
Palackého 1-3, 612 42 Brno
Czech Republic
Phone: +420 5 4156 2237

1: J Vet Med B Infect Dis Vet Public Health. 2002 Mar;49(2):111-4.
Hepatitis associated with herpes viral infection in the tortoise (Testudo horsfieldii). Hervas J, Sanchez-Cordon PJ, de Chacon Lara F, Carrasco L, Gomez-Villamandos JC. Departmento Anatomia Patologica Comparada, Facultad de Veterinaria, Universidad de Cordoba, Spain.

Herpesvirus infection in tortoises is largely characterized by the development of respiratory clinical signs. Usually lesions develop in the respiratory, oral pharyngeal, intestinal tract and are accompanied by cutaneous and ocular lesions. In chelonids affected by herpesvirus, systemic-type lesions in organs such as the liver and spleen are commonly observed. In this paper we describe a case of multifocal necrotic hepatitis associated with herpesviruses in an adult female land tortoise of the species Testudo horsfieldii. This article is the first description of a viral hepatitis in Testudo spp. with lesions compatible with herpesvirus infection, with no clinical signs or lesions in the respiratory system, oral cavity or other organs.

PMID: 12002420 [PubMed - indexed for MEDLINE]

1: Vet Pathol. 2000 Sep;37(5):377-85. Detection of chelonid herpesvirus DNA by nonradioactive in situ hybridization in tissues from tortoises suffering from stomatitis-rhinitis complex in Europe and North America. Teifke JP, Lohr CV, Marschang RE, Osterrieder N, Posthaus H. Bundesforschungsanstalt fur Viruskrankheiten der Tiere, Friedrich-Loeffler-Institute, Insel Riems, Germany.

Chelonid herpesvirus (ChHV) infection in tortoises associated with stomatitis-rhinitis complex is a severe, mostly epizootic disease characterized by proliferative and diphtheroid-necrotizing glossitis, pharyngitis, rhinitis, and tracheitis, often occurring with pneumonia and encephalitis. The UL5 gene from a German ChHV isolate was used to generate a digoxigenin-labeled 307-base-pair DNA probe by polymerase chain reaction (PCR). ChHV DNA was detected in paraffin-embedded tissues of five naturally infected tortoises (two Afghan tortoises [Testudo horsfieldii], USA; two Hermann's tortoises [Testudo hermanni], Switzerland; one T. hermanni, Germany) by means of in situ hybridization (ISH) and PCR. Distribution of ChHV DNA exhibits many characteristics of alphaherpesvirus but also some characteristics of betaherpesvirus infections. The amino acid sequence of a portion of the ChHV UL5 homolog exhibited more than 50% similarity to alphaherpesvirus UL5 proteins. Nuclear hybridization signals were detected in epithelial cells of the lingual mucosa and glands. Furthermore, ChHV DNA was observed in tracheal epithelium, pneumocytes, hepatocytes, the renal tubular epithelium, cerebral glia cells and neurons, and intramural intestinal ganglia. ChHV DNA in endothelial cells of many organs underlines the systemic character of the disease. Importantly, ChHV DNA was detected by ISH in multiple tissues of tortoises originating from different geographic provenances. This indicates a high degree of conservation of the UL5 gene fragment among viruses prevalent in tortoises on different continents. With the described ISH, a molecular biological tool is available for rapid and specific diagnosis of ChHV infections and, more importantly, comparative pathogenetic studies of ChHV isolates from geographically unrelated regions. 1: J Clin Microbiol. 2001 Sep;39(9):3156-63. Enzyme-linked immunosorbent assay for detecting herpesvirus exposure in Mediterranean tortoises (spur-thighed tortoise [Testudo graeca] and Hermann's tortoise [Testudo hermanni]). Origgi FC, Klein PA, Mathes K, Blahak S, Marschang RE, Tucker SJ, Jacobson ER. Department of Small Animal Clinical Sciences, University of Florida, Gainesville, Florida 32610-0126, USA. An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibodies to a herpesvirus associated with an upper respiratory tract disease in Mediterranean tortoises [spur-thighed tortoise (Testudo graeca) and Hermann's tortoise (Testudo hermanni)]. This serodiagnostic test was validated through a hyperimmunization study. The mean of the A(405) readings of the plasma samples collected at time zero of the hyperimmunization study plus three times the standard deviation was used as the cutoff for seropositivity in tortoises. ELISA results were compared to serum neutralization (SN) values for the same samples by using the McNemar test. The results obtained by SN and ELISA were not significantly different (P > 0.05). This new ELISA could be used as an important diagnostic tool for screening wild populations and private and zoo collections of Mediterranean tortoises. Publication Types: Evaluation Studies PMID: 11526144 [PubMed - indexed for MEDLINE] 1: Vet Pathol. 1999 Nov;36(6):624-7. Herpesvirus infection in tortoises (Malacochersus tornieri and Testudo horsfieldii). Une Y, Uemura K, Nakano Y, Kamiie J, Ishibashi T, Nomura Y. Laboratory of Veterinary Pathology, School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan. Large numbers of pancake tortoises (Malacochersus tornieri) and Horsfield tortoises (Testudo horsfieldii) in three consignments imported into Japan died soon after arrival. Some tortoises in the first consignment were dead on arrival. Postmortem examination of two of the pancake tortoises and four of the Horsfield tortoises revealed necrotizing lesions of the oral mucosa in both species, primarily in the tongue. Eosinophilic to amphophilic inclusion bodies were visible in the nuclei of mucosal epithelial cells in the lesions. Similar inclusion bodies were observed in the liver, spleen, adrenal glands, stomach, lungs, kidneys, small and large intestines, pancreas, and cerebrum of the pancake tortoises and in the liver, spleen, and pancreas of the Horsfield tortoises. Electron microscopic examination of the cells containing inclusion bodies showed herpesvirus-like particles about 100 nm in diameter in the cytoplasm. Nested polymerase chain reaction analysis using a herpesvirus consensus primer method confirmed the presence of a characteristic herpesvirus base sequence in tissue from these lesions.

PMID: 10568449 [PubMed - indexed for MEDLINE]


1: Mol Phylogenet Evol. 2002 Feb;22(2):174-83. Phylogenetic relationships among the species of the genus Testudo (Testudines: Testudinidae) inferred from mitochondrial 12S rRNA gene sequences. van der Kuyl AC, Ph Ballasina DL, Dekker JT, Maas J, Willemsen RE, Goudsmit J. Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands.

To test phylogenetic relationships within the genus Testudo (Testudines: Testudinidae), we have sequenced a fragment of the mitochondrial (mt) 12S rRNA gene of 98 tortoise specimens belonging to the genera Testudo, Indotestudo, and Geochelone. Maximum likelihood and neighbor-joining methods identify two main clades of Mediterranean tortoises, one composed of the species Testudo graeca, Testudo marginata, and Testudo kleinmanni and a second of Testudo hermanni, Testudo horsfieldii, and Indotestudo elongata. The first clade, but not the second, was also supported by maximum parsimony analysis. Together with the genus Geochelone, a star-like radiation of these clades was suggested, as a sister-group relationship between the two Testudo clades could not be confirmed. The intraspecies genetic variation was examined by sequencing the mt 12S rRNA fragment from 28 specimens of T. graeca and 49 specimens of T. hermanni from various geographic locations. Haplotype diversity was found to be significantly larger in T. graeca compared with T. hermanni, suggestive of reduced genetic diversity in the latter species, perhaps due to Pleistocene glaciations affecting northern and middle Europe or other sources of lineage reduction. No ancient mt 12S rRNA gene haplotypes were identified in T. graeca and/or T. hermanni originating from islands in the Mediterranean Sea, suggesting that these islands harbor tortoise populations introduced from the European and African mainland. (c)2002 Elsevier Science (USA).

PMID: 11820839 [PubMed - indexed for MEDLINE]


1: Arkh Anat Gistol Embriol. 1976 Aug;71(8):46-52. [Period of degeneration and features of the laminar distribution of the visual nerve endings in the midbrain tectum of the steppe turtle (Testudo horsfieldi, Gray)] [Article in Russian] Davydova TV.

A study of ultrastructural changes in the neuronal terminals of optic fibres at the level of tectum opticum (TO) in different periods after unilateral enucleation has revealed the process of degeneration in the optic system of Testudo horsfieldii as well as in Emysorbicularis L. to take a considerable time: from two weeks to six months. The character of destructive changes and the mode of utilization of desintegrated elements are dissimilar in different groups of retinal axons in the optic nerve and their terminals in TO. A comparison of the degeneration periods and the character of the changes in the optic fibres has made it possible to establish the following approximate correlations: all the nerve terminals of myelinated fibres degenerate after the "dark" type, whereas the terminals of non-myelinated axons are subjected to other kinds of transformations ("light", vesicular", neurofilamentous", "glycogen"). The analysis of the distribution of degenerating nerve terminals in the TO layers has demonstrated that there is a regularity in localization of different nerve terminals at certain levels of TO which is indicative of the layered organization of retino-tectile connections. Considerable differences in the neuronal and synaptic organization of TO have been revealed as well as in the character and the time of destructive changes of retino-tectile fibres after enucleation in T. horsfieldii and E. orbicularis.

PMID: 985095 [PubMed - indexed for MEDLINE]





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