Multicentric lymphoblastic lymphoma in a loggerhead sea turtle (Caretta caretta)

A juvenile female loggerhead sea turtle (Caretta caretta) stranded in Gran Canaria was submitted for necropsy. The turtle had exhibited anorexia and lethargy for 2 weeks prior to its death. At necropsy, the thymus was enlarged by two white and firm nodules. White nodules similar to those in thymus were observed in the plastron, thyroid gland, heart, aorta, left lung, spleen, liver, kidneys, stomach, and small intestine. Histopathology revealed a neoplastic proliferation of round cells identified as lymphoid cells. Ultrastructurally, the neoplastic cells were consistent with lymphoblastic cells, and viruses were not detected. The diagnosis was multicentric lymphoblastic lymphoma. This is the first report of a lymphoid neoplasm in a sea turtle.     

Morphologic and cytochemical characteristics of blood cells of juvenile loggerhead sea turtles (Caretta caretta)

A morphologic classification based on the cytochemical characteristics of blood cells of 35 juvenile loggerhead sea turtles (Caretta caretta) is described. Cytochemical stains included benzidine peroxidase, chloroacetate esterase, alpha-naphthyl butyrate esterase (with and without sodium fluoride), acid phosphatase (with and without tartaric acid), Sudan black B, periodic acid-Schiff, and toluidine blue. The morphologic characteristics of erythrocytes were similar to those reported in green turtles. Six types of white blood cells were identified: heterophils, eosinophils, basophils, lymphocytes, monocytes and thrombocytes. Except for the basophils, the rest of the white blood cells from loggerhead turtles had different cytochemical characteristics compared to blood cells from other sea turtle species. The leukocyte differential count was different from that reported for other sea turtle species. Heterophils were the most numerous leukocytes from these loggerhead turtles, followed by lymphocytes, eosinophils, monocytes and basophils. This paper provides a morphologic classification of blood cells of loggerhead sea turtles that is useful for veterinary surgeons involved in sea turtle conservation.     

Phacoemulsification of bilateral cataracts in a loggerhead sea turtle (Caretta caretta)

An immature free-living loggerhead sea turtle (Caretta caretta) of unknown sex was found moribund off the coast of Wise Point, Virginia. It was suffering from cachexia and had bilateral hypermature cataracts which were treated by phacoemulsification under general anaesthesia. The surgery restored the turtle's vision and it was returned to the wild.     

Computed tomography of the vertebral column and coelomic structures in the normal loggerhead sea turtle (Caretta caretta)

The aim of this study was to determine the normal computed tomography (CT) appearance of the vertebral column and coelomic structures of the loggerhead sea turtle (Caretta caretta) and to use three-dimensional (3D) and multiplanar reconstructions to indicate the position of each organ in relation to the vertebrae and carapace. Transverse sections of 1mm thickness were performed in seven clinically healthy and in five dead loggerhead sea turtles using multi-detector CT equipment. A computer workstation was used for multiplanar and 3D reconstructions. Dead turtles were frozen and sectioned in the transverse, dorsal and sagittal planes to compare the anatomical structures' appearance with CT images. Clinically relevant organs including the oesophagus, stomach, trachea, bronchi, lungs, liver, gallbladder, heart, spleen, kidneys and vertebral canal were identified in CT images. Computed tomography provides detailed information on the respiratory system and skeleton; the location of the coelomic structures with respect to the carapace and the vertebrae that is provided in this work will facilitate the use of other ancillary diagnostic techniques such as ultrasound, radiography and biopsy, thereby improving safety of access in surgical procedures.     

Chronic shoulder osteoarthritis in a loggerhead turtle (Caretta caretta)

This report describes the lesions seen in an extremely weak and emaciated adult male loggerhead turtle (Caretta caretta) found stranded near Perth, Western Australia. Blood collected from a dorsal cervical sinus immediately prior to euthanasia demonstrated severe anaemia, markedly elevated plasma concentrations of creatinine kinase (2263 U/L), alkaline phosphatase (58 U/L), urea (18.1 mmol/L) and hypoglycaemia (glucose 0.7 mmol/L). Necropsy examination demonstrated a severe chronic osteoarthritis of the right shoulder joint with marked remodelling of the glenoid fossa, coracoid, scapula and humerus. There was marked synovial effusion and periarticular fibrosis. Cytological examination and culture of synovial fluid failed to demonstrate an infectious agent. Other findings were low numbers of trypanorhynch cysts present on the surface of the liver and numerous spirorchid eggs in the attached mesentery. Several large and small spirorchid flukes were present in the heart and there was a localised endarteritis in the left aortic arch associated with this infection. Numerous nematode eggs and at least three species of spirorchid eggs were seen microscopically in faeces collected from the terminal colon. The pathogenesis and significance of these lesions is discussed.     

Ultrasonographic imaging of loggerhead sea turtles (Caretta caretta)

Twenty live and five dead juvenile and subadult loggerhead sea turtles were examined ultrasonographically. Ten soft tissue areas of the integument were used as acoustic windows: cervical-dorsal and cervical-ventral, left and right cervicobrachial, left and right axillary, left and right prefemoral and left and right postfemoral windows. Anatomical cross-sections were performed on the dead turtles to provide reference data. The fourth and fifth cervical vertebrae, the spinal cord, and the venous sinuses of the external jugular vein were clearly visible through the cervical-dorsal acoustic window, and the oesophagus and the heart were imaged through the cervical-ventral acoustic window. The stomach was more frequently visible through the left axillary acoustic window. The liver could be imaged through both sides, but the right axillary acoustic window was better for visualising the gall bladder. The large and small intestines and the kidneys were visible through the right and left prefemoral acoustic windows; the kidneys were easily identified by their intense vasculature.     

Intestinal candidiasis in a loggerhead sea turtle (Caretta caretta): an immunohistochemical study

Post mortem examination of a juvenile loggerhead sea turtle (Caretta caretta) stranded in the Canary Islands revealed a fishing-line in the small intestine. Histologically, severe necrotic enteritis, multiple haemorrhages, and marked oedema of the intestinal submucosa were observed. Yeast cells and fungal hyphae were seen in the lamina propria of the intestinal mucosa and in the connective tissue of the submucosa. Because fungal cultures were not taken at the time of necropsy, an immunohistochemical study was performed in order to identify the fungus involved. Specific monoclonal and heterologously absorbed polyclonal antibodies served as the primary reagents for identification of aspergillosis, candidiasis, fusariosis, geotricosis, scedosporiosis, and zygomycosis, using an indirect immunofluorescence staining technique. The fungal elements were strongly stained only by a polyclonal antibody against Candida albicans and a monoclonal antibody against C. albicans. There are no known previous reports of Candida sp. causing skin disease or systemic mycotic infection in sea turtles.     

Evaluation of Doppler ultrasonography for the measurement of blood flow in young loggerhead sea turtles (Caretta caretta)

The aim of this study was to identify ultrasound accessible blood vessels in the loggerhead sea turtle (Caretta caretta) and describe their Doppler waveform patterns, peak systolic velocity, mean velocity, systolic/diastolic ratio as well as pulsatility and resistive indices. B-mode, colour and pulsed-wave Doppler examinations were performed on 10 turtles. Flow measurements were recorded for the left and right aorta, epigastric and internal iliac arteries, and right hepatic vein. Additionally, the large blood vessels of three dead turtles were injected with latex and dissected for anatomical support. A parabolic flow velocity profile was observed in all arteries. The waveforms of the right and left aortic arteries showed an unusual pattern when compared with mammals. The hepatic vein flow velocity waveform of the loggerhead sea turtle was found to be similar to that in the dog, although the flow velocity in the C-wave was higher than that in the A-wave. The low resistance flow pattern observed suggests that the loggerhead sea turtle's organs require a continuous blood supply.     

Pharmacokinetics of ticarcillin in the loggerhead sea turtle (Caretta caretta) after single intravenous and intramuscular injections.

Three captive loggerhead sea turtles, Caretta caretta, were used in four trials, one i.v. and three i.m., to determine the pharmacokinetic properties of a single dose of ticarcillin. For the i.v. study, each turtle received a single 50 mg/kg dose and blood samples were collected at 0, 0.5, 1, 2, 4, 6, 8, and 12 hr and at 1, 1.5, 2, 2.5, 3, 4, 6, 8, 10, and 14 days after administration. For the i.m. study, each turtle received one of three dosages (25, 50, or 100 mg/kg) in a randomized complete block design and blood samples were collected at the same time intervals. Each trial was separated by a minimum of 28 days to allow for complete drug clearance. Drug concentration in plasma was determined by a validated liquid chromatography-mass spectrometry assay. For the i.v. study, the elimination half-life was 5.0 hr. The apparent volume of distribution and plasma clearance were 0.17 L/kg and 0.0218 L/hr/kg, respectively. For the i.m. study, mean time to maximum plasma concentrations ranged from 1.7 ( +/- 0.58) hr in the 50 mg/kg group to 3.7 (+/- 2.5) hr in the 100 mg/kg group. Mean bioavailability ranged from 45% ( +/- 15%) in the 50 mg/kg group to 58% (+/- 12%) in the 100 mg/kg group, and the mean residence time ranged from 7.5 ( +/- 2.6) hr in the 25 mg/kg group to 16 (+/- 6.8) hr in the 100 mg/kg group. Two turtles had slight alanine aminotransferase elevations that were not clinically apparent at two different dosages, but otherwise, blood chemistries were unaffected. Possible i.m. dosage regimens for loggerhead sea turtles are 50 mg/kg q24 hr or 100 mg/kg q48 hr. Liver enzymes should be monitored during treatment.     

Proliferative dermatitis in a loggerhead turtle, Caretta caretta, and a green turtle, Chelonia mydas, associated with novel papillomaviruses

A subadult loggerhead turtle, Caretta caretta, presented with generalized small, white, raised lesions over its neck, shoulders, and all four flippers. A juvenile green turtle, Chelonia mydas, recently treated for fibropapillomatosis, presented with four similar localized lesions on one flipper. To diagnose the conditions, biopsies of the lesions were taken for histopathology, electron microscopy, and molecular diagnostics. Histopathologic findings were similar in the two turtles and skin lesions were characterized by multifocal areas of epidermal hyperplasia accompanied by variation and abnormalities in the nuclear morphology of keratinocytes and a few intranuclear inclusions in some cells. Transmission electron microscopy revealed multiple epithelial cells with large intranuclear aggregates of virions consistent in morphology with papillomavirus. Papillomavirus was detected in samples from both turtles by polymerase chain reaction (PCR). Sequence analysis of the partial sequence of the papillomavirus E1 gene revealed two viruses (CcPV and CmPV) that were distinct from each other and from other species in Papillomaviridae, and likely represent two novel species and perhaps a new genus.     

Ultrastructural characteristics of blood cells of juvenile loggerhead sea turtles (Caretta caretta)

Ultrastructural characteristics of erythrocytes, heterophils, eosinophils, lymphocytes, monocytes and thrombocytes of the loggerhead sea turtle (Caretta caretta) were evaluated, using blood samples from 15 healthy juvenile animals. Except for the eosinophils, the rest of the white blood cells from loggerhead turtles had similar ultrastructural characteristics compared with blood cells from other sea turtle species. Eosinophils from loggerhead turtles were homogeneous in size, and no crystalline structures were observed within the granules. This paper provides an ultrastructural characterization of blood cells of loggerhead sea turtles, as a reference for future haematological studies of this species.     

Sectional anatomic and magnetic resonance imaging features of the head of juvenile loggerhead sea turtles (Caretta caretta)

Objective-To compare anatomic features of cross-sectional specimens with those of MRI images of the heads of loggerhead sea turtles (Caretta caretta). Animals-5 cadavers of juvenile female loggerhead sea turtles. Procedures-Spin-echo T1-weighted and T2-weighted MRI scans were obtained in sagittal, transverse, and dorsal planes with a 0.2-T magnet and head coil. Head specimens were grossly dissected and photographed. Anatomic features of the MRI images were compared with those of gross anatomic sections of the heads from 4 of these turtles. Results-In the MRI images, anatomic details of the turtles' heads were identified by the characteristics of signal intensity of various tissues. Relevant anatomic structures were identified and labeled on the MRI images and corresponding anatomic sections. Conclusions and Clinical Relevance-The MRI images obtained through this study provided valid information on anatomic characteristics of the head in juvenile loggerhead sea turtles and should be useful for guiding clinical evaluation of this anatomic region in this species.     

Atypical ocular Chelonoid herpesvirus manifestations in a captive Loggerhead turtle (Caretta caretta)

A captive loggerhead turtle (Caretta caretta) of unknown sex, 3 years of age, presented with bilateral mucoid secretions, severe chemosis, conjunctival hyperemia, and globe retraction. The animal was evaluated ophthalmologically and systemically, and hematological, microbiological, and conjunctival cytological and biopsy samples were collected for complementary diagnosis. The histopathological examination showed amphophilic intranuclear inclusions associated with severe inflammatory infiltrate. The diagnosis of Chelonid alphaherpesvirus 5 (ChAHV 5) was confirmed with end point PCR. Following systemic treatment with L‐lysine, acyclovir and vitamin A, the ocular signs resolved. No amphophilic intranuclear inclusions were seen in a follow‐up biopsy 5 months later, and there has been no recurrence of clinical ophthalmic signs during a 4‐year follow‐up. It is suggested that ChAHV 5 be considered as a differential diagnosis in captive marine turtles that present for conjunctival disease other than fibropapillomatosis.     

Comparative study of hematologic and plasma biochemical variables in Eastern Atlantic juvenile and adult nesting loggerhead sea turtles (Caretta caretta)

Background: Plasma biochemical and hematologic variables are important in the management of endangered sea turtles, such as loggerheads. However, studies on blood biochemistry and hematology of loggerheads are limited, and different concentrations according to variable criteria have been reported. Objective: The purpose of this study was to establish and compare baseline plasma chemistry and hematology values in Eastern Atlantic juvenile and adult nesting loggerhead sea turtles (Caretta caretta). Methods: Blood samples were collected from 69 healthy juvenile loggerhead sea turtles after their rehabilitation in captivity, and from 34 adult nesting loggerheads after oviposition. Fresh blood was used for leukocyte differential count and PCV determination. Heparinized blood was used for RBC and WBC counts. Plasma biochemical concentrations were measured using an automated biochemical analyzer. For the comparative study, nonparametric statistical analysis was done using the Mann–Whitney U‐test. Results: Minimum, maximum, and median concentrations were obtained for 14 hematologic and 15 plasma chemistry variables. Statistically significant differences between juvenile and adult turtles were found for PCV; RBC, WBC, and leukocyte differential counts; total protein, albumin, globulins, calcium, triglycerides, glucose, total cholesterol and urea concentrations; and lactate dehydrogenase activity. Conclusions: Age, size, and reproductive status cause important variations in the hematologic and plasma biochemical results of loggerheads. The reference values obtained in this study may be used as a standard profile, useful for veterinary surgeons involved in sea turtle conservation.     

Ingesta passage and gastric emptying times in loggerhead sea turtles (Caretta caretta)

Ingesta passage times of soft flat foam dishes and gastric emptying time of barium-impregnated polyethylene spheres (BIPS) were measured in 22 and 8 loggerhead sea turtles (Caretta caretta), respectively. Transit time (T(1)) was considered as the time between ingestion and first elimination, and retention time (T(50)) and total transit time (T(85)) the expulsion time of 50% and 85% of the markers, respectively. The experiments were carried out at different times of the year and water temperature was recorded. A set of dorso-ventral radiographs was taken to locate the BIPS, and the gastrointestinal anatomy of 5 dead turtles was studied to help with interpretation of the radiographs. No significant correlation was observed between T(1), T(50), T(85) and minimum straight carapace length (SCLmin) or body mass and no statistical difference was found in ingesta passage transit times between juvenile (n = 6) and sub-adult turtles (n = 16). Mean passage times of the dishes (in days) were: T(1) = 9.05, T(50) = 12.00 and T(85) = 13.19. Gastric emptying time using BIPS was 24-48 h. The transit time (T(1)) for the BIPS was longer (13.25 +/- 4.86 days) than the foam markers (8.5 +/- 2.73 days) in 8 turtles studied simultaneously. Although the total transit time tended to be faster in turtles submitted to water temperatures between 20 degrees C and 23.6 degrees C no significant correlation was observed between T(1), T(50) and T(85) and the temperature.     

Radiographic features of the limbs of juvenile and subadult loggerhead sea turtles (Caretta caretta)

This study aimed to provide the normal radiographic anatomic appearance of the limbs of the loggerhead sea turtle, Caretta caretta. Dorsopalmar and dorsoplantar radiographs were taken of the forelimbs and hindlimbs of 15 juvenile and 15 subadult loggerhead sea turtles, 17 alive and 13 dead. For comparison, computed tomographic, gross anatomic, osteologic, and histologic studies were performed on the limbs of 5 of the sea turtles. Bones from the distal part of the fore and hind flippers were seen in detail with a mammographic film-screen combination. The pectoral and pelvic girdles, superimposed by the carapace, were better seen on standard radiographs with the use of rare-earth intensifying screens. Mammographic radiographs of the manus of 5 small juvenile turtles showed active growth zones. Visualization of bone contours in the distal part of the limbs was clearer than in mammals owing to the very few superimpositions. The presence of a substantial amount of cartilage in the epiphyses produced better visibility of limb ends. We conclude that use of a mammography film-screen combination is the best way to evaluate the bony and joint structures of the limbs of sea turtles. Radiography provides reliable images for clinical purposes. Considering the low cost and logistics of this technique, it is a practical ancillary test for marine animal rehabilitation centers to use.