Enzyme-linked immunosorbent assay for detecting herpesvirus exposure in Mediterranean tortoises (spur-thighed tortoise [Testudo graeca] and Hermann's tortoise [Testudo hermanni])

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.     

Epidemiology and biology of nematodofauna affecting Testudo hermanni, Testudo graeca and Testudo marginata in Italy

Tortoises of the genus Testudo living in Italy are Testudo hermanni, Testudo graeca and Testudo marginata. Although a great deal of information has been acquired on the internal medicine and surgery on these animals, little is known of their parasitological fauna. A survey on the presence of gastro-intestinal parasitic nematodes in tortoises bred in Italy was carried out to acquire extensive epidemiological data. Stool samples of 62 tortoises (37 T. hermanni hermanni, 13 T. graeca, 6 T. hermanni boettgeri and 6 T. marginata) were macroscopically examined and subjected to qualitative (flotation and Baermann techniques) and quantitative (McMaster technique) microscopic examinations. Adult nematodes were identified using morphological keys. Eggs of oxyurids (species indistinguishable) and adults of Alaeuris numidica, Mehdiella microstoma, Mehdiella uncinata, Tachygonetria longicollis, Tachygonetria conica and Tachygonetria palearcticus (Oxyurida, Pharyngodonidae); larvae and adults of Atractis dactyluris (Ascaridida, Atractidae); and eggs and adults of Angusticaecum holopterum (Ascaridida, Ascarididae) were identified in all animals >1 year, whereas all tortoises <1 year of age were parasite-free. Moreover, this study reports for the first time A. numidica and M. microstoma in T. marginata. This work shows that nematodes affecting tortoises are diffused in Italy and highlights the need to investigate their epidemiology more in depth.     

Pancreatoblastoma--histopathological and ultrastructural analysis of two cases.

Pancreatoblastoma has been described in children and characterized by unique histologic features and excellent clinical course. Ultrastructural and immunohistochemical studies of pancreatoblastoma reveal either exocrine alone or both endocrine and exocrine differentiation. We present two cases of pancreatoblastoma in children in which immunohistochemical and ultrastructural examination failed to demonstrate features of either enzyme or hormone production and which became worse in clinical course. We assume that pancreatoblastomas are tumors which differentiate more toward acinar or ductal elements than toward islet cell.     

Efficacy of oxfendazole and fenbendazole against tortoise (Testudo hermanni) oxyurids

Thirty-six tortoises (Testudo hermanni) with naturally acquired oxyurids infections were used to assess the anthelmintic efficacy of oxfendazole (Dolthene; Merial) and fenbendazole (Panacur; Hoechst Roussel Vet). Animals were randomly assigned to three groups (A, B, and C) based on sex and weight. Animals in group A (seven males and six females) were orally treated with oxfendazole at dose rate of 66 mg/kg, group B animals (nine males and eight females) were orally treated with fenbendazole at dose rate of 100 mg/kg, and group C animals (three males and three females) were not treated and served as controls. All animals were individually stabled in plexiglas boxes under controlled conditions of temperature, humidity, and light beginning 7 days pretreatment and continuing for the duration of the trial. Individual tortoises feces were examined daily by the McMaster technique and drugs efficacy was assessed by the fecal eggs count reduction (FECR) test. Both drugs showed 100% of FECR. However, oxfendazole reached this level 12 days after treatment, whereas 31 days after treatment were necessary to obtain the same stable result with fenbendazole. The two drugs were well tolerated by all the animals and no adverse reactions were observed after treatment.     

The distribution of dopamine in the brain of a tortoise, Geochelone chilensis (Gray)

1. Dopamine was found in the tortoise brain; the highest concentration (3.7 mug/g) occurred in a region of the telencephalon containing the nucleus basalis and cortex olfactoria. This region may be considered similar to the mammalian and avian corpus striatum with respect to dopamine.2. The administration of reserpine or prenylamine substantially decreased the concentration of dopamine in the tortoise brain. This effect appeared later and lasted longer than in birds or mammals. When these drugs were given in doses which nearly depleted brain dopamine the characteristic signs observed in birds or mammals were not present.     

Activity of vagal afferent fibers innervating CO2-sensitive receptors in the tortoise, Testudo hermanni

Experiments were done on the tortoise, Testudo hermanni (anesthetized), or on its lung-vagus preparation in which all other internal organs had been removed. By recording the afferent impulse patterns in fine strands of the vagus, three kinds of CO2-sensitive receptors were demonstrated. Heymans-type chemoreceptors: Afferents originating from thoracic arterial chemoreceptors fired randomly at amplitudes below 100 microV. Firing frequency increased when the animal was ventilated with CO2-enriched, nitrogen or hypoxic mixtures, or was given NaCN. Intrapulmonary CO2 receptors (IPC). In an artificially ventilated tortoise and in lung-vagus preparations where the lung CO2-fraction (FCO2) was below 2.5%, single units with a spike amplitude above 200 microV fired regularly. When CO2-enriched air was inspired, impulse frequency decreased markedly, and when the lung was washed with air, it returned to the control frequency following the initial excitation. In lung-vagus preparations the impulse frequency did not change with lung inflation, was null for FCO2 greater than 2.5-3.5%, and was inversely proportional to lower FCO2's. Intrapulmonary CO2-sensitive mechanoreceptors: In anesthetized artificially ventilated tortoises, impulse bursts synchronized with inspiration decreased when the animal inspired 5 or 10% CO2 in air. In lung-vagus preparations, slowly adapting mechanoreceptor responses provoked by lung inflation were reduced when the lung was inflated with CO2-enriched air.     

Comparative ultrastructural morphometric analysis of atrial specific granules in the bat,mouse, and rat

Size, incidence, and volume density of atrial specific granules (ASG) in right atrial cells from five animals each of the rat (average weight 210 g), mouse (average weight 28 g), fruit-eating bat Megaloglossus woermanni (BMW;average weight 35 g), and the insect-eating bat Pipistrellus pipistrellus (BPP; average weight 6 g) have been compared via ultrastructural morphometry. In all three parameters of granule measurement, significantly higher figures were obtained in the rodents than in the bats. However, between the rat and the mouse, as also between the two species of bats, no significant differences were noted in any of the measurements. These results therefore do not support the prevalent view that the number and size of the granules decrease with increase in size of the animal species. The low content of ASG in atrial cells of the bats is probably an indication of low demand for the natriuretic hormone of the granule, because, in such animals, and particularly in flight, conservation of fluid and electrolytes is of paramount importance. This suggests that granule content is adapted to fluid and electrolyte regulation in relation to the functional capacity of the animal. We also observed ASG-like sgtructures in endothelial cells of capillaries of bat tissue but not in rodents. The function of these granules or whether or not they represent atrial specific ones is not clear from the present study. © Wiley-Liss, Inc.     

Coexistence of two lymphomas with distinctive histologic, ultrastructural, and immunologic features.

The unusual coexistence of two distinct lymphomas in 44-year-old woman is described. Nodular, poorly differentiated lymphocytic lymphoma and diffuse histiocytic lymphoma were present in separate sites and were readily distinguished both histologically and ultrastructurally. In addition, the lymphocytic lymphoma was shown to be derived from complement receptor B lymphocytes of follicular center cell type, whereas the histiocytic lymphoma cells were devoid of complement receptors, receptors for IgG (Fc receptors), and surface immunoglobulin. Despite intensive chemotherapy and radiation therapy, the patient died within eight months of the initial diagnosis. Although histiocytic lymphoma was widely disseminated at autopsy, lymphocytic lymphoma was not found. Presumably the histiocytic lymphoma was refractory to therapy, in contrast to the lymphocytic lymphoma, which was selectively eradicated.     

Mucinous cystadenoma of the lung. A report of two cases with immunohistochemical and ultrastructural analysis.

We describe two patients who presented with solitary pulmonary masses that consisted of unilocular cysts lined by columnar mucinous epithelium. The cysts contained copious mucus. The epithelial lining of the cysts showed foci of stratification and papillary infolding. Histologically identical lesions have previously been termed unusual mucous cysts or mucinous cystadenomas. We believe that these tumors are true neoplasms differentiating toward the respiratory epithelial mucous cell. They should be distinguished from a variety of pulmonary neoplasms including bronchoalveolar carcinoma, bronchial mucous gland adenoma, mucoepidermoid carcinoma, and metastatic adenocarcinoma.     

A human adult Wilms' tumor. Histologic, ultrastructural, and cytogenetic analysis

The cytogenetic, histologic, and electron microscopic studies of an adult patient with Wilms' tumor are presented. Wilms' tumor (nephroblastoma) is a common renal tumor of childhood but is extremely rare in people over 15 years old. The histologic analysis of the patient's tumor, including both light and electron microscopic analysis, indicated that this tumor satisfies the histologic criteria for an adult Wilms' tumor, namely, blastemic cells that are immature renal parenchymal cells, embryonic tubular structures, and a scanty stromal component consisting of loosely arranged spindle cells. The tumor showed several ultrastructural features characteristic of adult Wilms' tumor, namely, markedly elongated mitochondria, autophagic vacuoles, and intracytoplasmic filaments. Karyotypic analysis was performed on the patient's peripheral leukocytes and tumor cells. The leukocytes showed no significant increase in gaps and breaks, and the patient appears to have a normal male karyotype. Some interesting chromosomal anomalies were observed in the cultured tumor cells: at least one chromosome 13, both chromosomes 22, and the X chromosome are missing, three markers are present, and there is a possible deletion of 12p.     

Myxoid liposarcoma. An ultrastructural study of two cases.

Two cases in which myxoid liposarcomas were examined by use of light and electron microscopy are presented. The relationship of this tumor to the development of normal adipose tissue is discussed. Most of the ultrastructural features were identical to those of developing white adipose tissue. Features not typical of developing white adipose tissue, such as congeries of 60-Angstrom filaments and large masses of glycogen, are also illustrated and discussed.     

Quantitative ultrastructural analysis of coronary atherosclerotic involvement in two macaque species

Ultrastructural analyses were employed to observe and to compare in detail lesions of the coronary artery of cynomolgus and rhesus monkeys. Animals were fed individually with the same atherogenic ration under identical conditions for 4, 8, and 12 months, and controls of each species were fed with a low fat, cholesterol-free ration. Transmission electron microscopic studies of coronary arteries from these animals led to the following conclusions: (1) Synthetic smooth muscle cells (SMC) without lipid and macrophages without lipid appeared more frequently in the cynomolgus lesions than in the rhesus lesions. Furthermore, phenotypic expression of synthetic SMCs in the cynomolgus was more active with greater diversity, while the rhesus showed less phenotypic modulation. Macrophages without lipid appeared frequently in the cynomolgus media. (2) Increased percentages of both synthetic SMCs with lipid and macrophages with lipid were demonstrated in the cynomolgus lesions as compared to those in the rhesus. This indicates that foam cells, including SMC- and macrophage-derived foam cells, are more prevalent in cynomolgus than in rhesus. They are considered to play an important role in atherogenesis. (3) Medial disruption, synthetic SMCs, and macrophages containing lipid appeared more often in cynomolgus media than in rhesus media. (4) There were greater percentages of both synthetic SMCs and macrophages in the intima of the myocardial side of coronary arteries in both species. (5) Approximately 42% of all foam cells in the cynomolgus lesions were derived from SMCs. There were fewer macrophages in rhesus lesions. (6) The difference in expression between the two macaque species reflects different responses of macrophages to medial smooth muscle cell (SMC) components. The configuration of the artery wall could be one of the important indicators of these different expressions.     

Leptoconops bezzii (Diptera: Ceratopogonidae) parasitizing tortoises Testudo graeca (Testudines: Testudinidae) in mountain ranges of Lebanon and western Syria

Biting midges of the genus Leptoconops, subgenus Leptoconops (Diptera: Ceratopogonidae) were repeatedly found parasitizing on spur-thighed tortoises Testudo graeca in western Syria and Lebanon. Collected females were assigned to the species L. bezzii according to their morphological characteristics. Tortoises parasitized by midges were observed throughout the daytime, under mild temperatures, and slight winds, in Mediterranean or stony steppe habitats. Intensity of the infestations ranged from a few specimens up to thousands of midges per tortoise. Midges infested predominantly the tortoises’ carapax, finding their approach to the capillaries in the seams between the keratinized scutes. The vector potential of ceratopogonids is discussed.