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.     

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 coelomic structures of loggerhead sea turtles

OBJECTIVE: To compare cross-sectional anatomic specimens with images obtained via magnetic resonance imaging (MRI) of the coelomic structures of loggerhead sea turtles (Caretta caretta). ANIMALS: 5 clinically normal live turtles and 5 dead turtles. PROCEDURES: MRI was used to produce T1- and T2- weighted images of the turtles, which were compared with gross anatomic sections of 3 of the 5 dead turtles. The other 2 dead turtles received injection with latex and were dissected to provide additional cardiovascular anatomic data. RESULTS: The general view on the 3 oriented planes provided good understanding of cross-sectional anatomic features. Likewise, major anatomic structures such as the esophagus, stomach, lungs, intestine (duodenum and colon), liver, gallbladder, spleen, kidneys, urinary bladder, heart, bronchi, and vessels could be clearly imaged. It was not possible to recognize the ureters or reproductive tract. CONCLUSIONS AND CLINICAL RELEVANCE: By providing reference information for clinical use, MRI may be valuable for detailed assessment of the internal anatomic structures of loggerhead sea turtles. Drawbacks exist in association with anesthesia and the cost and availability of MRI, but the technique does provide excellent images of most internal organs. Information concerning structures such as the pancreas, ureters, intestinal segments (jejunum and ileum), and the reproductive tract is limited because of inconsistent visualization.     

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.     

Relationship between barnacle epibiotic load and hematologic parameters in loggerhead sea turtles (Caretta caretta), a comparison between migratory and residential animals in Pamlico Sound, North Carolina.

Health status of a total of 57 loggerhead sea turtles (Caretta caretta; 42 migratory and 15 residential turtles) was analyzed using body condition and hematologic parameters. A subset of 18 juvenile migratory loggerhead sea turtles in the fall of 1997 and 15 residential turtles in the summer of 2000 were analyzed for barnacle epibiota. The migratory group had significantly higher red blood cell counts and percent heterophils and significantly lower percent lymphocyte and absolute eosinophil counts, as well as significantly lower plasma concentrations of calcium, sodium, chloride, potassium, glucose, alkaline phosphatase, and anion gap. Many of these variations may be because of physiology of migration. A positive association between turtle weight and hematocrit was detected and may be because of larger turtles diving for longer periods of time. There were no significant differences of epibiota load, health of the turtles, or condition index between turtles captured during the two events.     

Evaluation of hematology and serum biochemistry of cold-stunned green sea turtles (Chelonia mydas) in North Carolina, U.S.A

Hypothermia or cold-stunning is a condition in which the body temperature of an animal decreases below normal physiologic range and which has been linked to severe morbidity in sea turtles. Reports have focused on the physiologic changes caused by cold-stunning in Kemp's Ridley sea turtles (Lepidochelys kempii) and loggerhead sea turtles (Caretta caretta), but few have evaluated the green sea turtle (Chelonia mydas). This study evaluated hematologic and serum biochemical profiles of cold-stunned green sea turtles in North Carolina, USA. When compared with healthy, free-ranging juvenile green turtles from the same region, cold-stunned turtles exhibited hypoglycemia, hypocalcemia (both total and ionized calcium), hyponatremia, hypokalemia, hypoproteinemia, hypoalbuminemia, hyperphosphatemia, and elevations in uric acid and blood urea nitrogen. These findings contrast with some previously reported changes in cold-stunned Kemp's Ridley and loggerhead sea turtles. These results emphasize the importance of basing therapeutic regimens on biochemical analyses in cold-stunned sea turtles.     

Intraocular pressure of juvenile loggerhead sea turtles (Caretta caretta) held in different positions.

The intraocular pressure of 12 apparently healthy juvenile loggerhead sea turtles (Caretta caretta) was determined by applanation tonometry while the turtles were held in dorsoventral, ventrodorsal, and head-down suspended positions. The median intraocular pressures were 5 mmHg (range 4 to 9 mmHg) in the dorsoventral position, 7 mmHg (range 5 to 12 mmHg) in the ventrodorsal position, and 23 mmHg (range 17 to 33 mmHg) in the suspended position.     

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.     

Evaluation of five clinical chemistry analyzers for use in health assessment in sea turtles

OBJECTIVE: To compare blood biochemical values obtained from a handheld analyzer, 2 tabletop analyzers, and 2 diagnostic laboratories by use of replicate samples of sea turtle blood. DESIGN: Validation study. ANIMALS: 22 captive juvenile sea turtles. PROCEDURES: Sea turtles (18 loggerhead turtles [Caretta caretta], 3 green turtles [Chelonia mydas], and 1 Kemp's ridley turtle [Lepidochelys kempii]) were manually restrained, and a single blood sample was obtained from each turtle and divided for analysis by use of the 5 analyzers. Hematocrit and concentrations or activities of aspartate aminotransferase, creatine kinase, glucose, total protein, albumin, BUN, uric acid, P, Ca, K, Na, Cl, lactate dehydrogenase, and alkaline phosphatase were determined. Median values for each analyte were compared among the analyzers. RESULTS: Significant differences were found among the analyzers for most values; however, data obtained from the 2 diagnostic laboratories were similar for all analytes. The magnitude of difference between the diagnostic laboratories and in-house units was > or = 10% for 10 of the 15 analytes. CONCLUSIONS AND CLINICAL RELEVANCE: Variance in the results could be attributed in part to differences in analyzer methodology. It is important to identify the specific methodology used when reporting and interpreting biochemical data. Depending on the variable and specific case, this magnitude of difference could conceivably influence patient management.     

Successive changes of hematologic characteristics and plasma chemistry values of juvenile loggerhead turtles (Caretta caretta).

Hematologic characteristics and plasma chemistry values of juvenile loggerhead turtles (Caretta caretta) from the ages of 1 mo to 3 yr were obtained to establish baseline values. Five clinically normal loggerhead turtles were selected from the same clutch and raised in an indoor artificial nesting beach. Blood samples were successively collected and examined for various blood characteristics for a maximum total of 15 times. Hematologic characteristics, including packed cell volume, white blood cell counts, and white blood cell differentials; and plasma chemistry values, including total bilirubin, total protein, albumin, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, gamma-glutamic transpeptidase, creatinine, blood urea nitrogen, uric acid, alkaline phosphatase, amylase, triglyceride, total cholesterol, ionized sodium, ionized potassium and ionized chlorine, were measured. These results were used to establish a hematology and blood chemistry baseline for captive juvenile loggerhead turtles and will aid in their medical management.     

Plasma concentrations of praziquantel after oral administration of single and multiple doses in loggerhead sea turtles (Caretta caretta)

OBJECTIVE: To determine the pharmacokinetics of praziquantel following single and multiple oral dosing in loggerhead sea turtles. ANIMALS: 12 healthy juvenile loggerhead sea turtles. PROCEDURE: Praziquantel was administered orally as a single dose (25 and 50 mg/kg) to 2 groups of turtles; a multiple-dose study was then performed in which 6 turtles received 3 doses of praziquantel (25 mg/kg, PO) at 3-hour intervals. Blood samples were collected from all turtles before and at intervals after drug administration for assessment of plasma praziquantel concentrations. Pharmacokinetic analyses included maximum observed plasma concentration (Cmax), time to maximum concentration (Tmax), area under the plasma praziquantel concentration-time curve, and mean residence time (MRTt). RESULTS: Large interanimal variability in plasma praziquantel concentrations was observed for all dosages. One turtle that received 50 mg of praziquantel/kg developed skin lesions within 48 hours of administration. After administration of 25 or 50 mg of praziquantel/kg, mean plasma concentrations were below the limit of quantification after 24 hours. In the multiple-dose group of turtles, mean plasma concentration was 90 ng/mL at the last sampling time-point (48 hours after the first of 3 doses). In the single-dose study, mean Cmax and Tmax with dose were not significantly different between doses. After administration of multiple doses of praziquantel, only MRTt was significantly increased, compared with values after administration of a single 25-mg dose. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of 25 mg of praziquantel/kg 3 times at 3-hour intervals may be appropriate for treatment of loggerhead sea turtles with spirorchidiasis.     

Computed tomographic anatomy of the head of the loggerhead sea turtle (Caretta caretta)

The heads of three loggerhead sea turtles were disarticulated and imaged immediately to minimize postmortem changes and then frozen and sectioned. For computed tomography (CT) imaging, the heads were positioned in ventral recumbency. Transverse CT images with soft-tissue window were obtained from the olfactory sac region to the temporomandibular joint region. After CT imaging, the heads were sectioned and the gross sections were compared to CT images, to assist in the accurate identification of the anatomic structures. Different clinically relevant anatomic structures were identified and labelled in two series of photographs (CT images and anatomic cross-sections). CT images provided good differentiation between the bones and the soft tissues of the head. The information presented in this paper should serve as an initial reference to evaluate CT images of the head of the loggerhead sea turtle and to assist in the interpretation of lesions of this region.     

Medetomidine,ketamine, and sevoflurane for anesthesia of injured loggerhead sea turtles: 13 cases (1996-2000)

OBJECTIVE: To determine safety and efficacy of an anesthetic protocol incorporating medetomidine, ketamine, and sevoflurane for anesthesia of injured loggerhead sea turtles. DESIGN: Retrospective study. ANIMALS: 13 loggerhead sea turtles. PROCEDURE: Anesthesia was induced with medetomidine (50 microg/kg [22.7 microg/lb], IV) and ketamine (5 mg/kg (2.3 mg/lb], IV) and maintained with sevoflurane (0.5 to 2.5%) in oxygen. Sevoflurane was delivered with a pressure-limited intermittent-flow ventilator. Heart rate and rhythm, end-tidal partial pressure of CO2, and cloacal temperature were monitored continuously; venous blood gas analyses were performed intermittently. Administration of sevoflurane was discontinued 30 to 60 minutes prior to the end of the surgical procedure. Atipamezole (0.25 mg/kg [0.11 mg/lb], IV) was administered at the end of surgery. RESULTS: Median induction time was 11 minutes (range, 2 to 40 minutes; n = 11). Median delivered sevoflurane concentrations 15, 30, 60, and 120 minutes after intubation were 2.5 (n = 12), 1.5 (12), 1.25 (12), and 0.5% (8), respectively. Heart rate decreased during surgery to a median value of 15 beats/min (n = 11). End-tidal partial pressure of CO2 ranged from 2 to 16 mm Hg (n = 8); median blood gas values were within reference limits. Median time from atipamezole administration to extubation was 14 minutes (range, 2 to 84 minutes; n = 7). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a combination of medetomidine and ketamine for induction and sevoflurane for maintenance provides safe, effective, controllable anesthesia in injured loggerhead sea turtles.     

Pharmacokinetics of florfenicol in loggerhead sea turtles (Caretta caretta) after single intravenous and intramuscular injections.

The pharmocodynamics of single injections of florfenicol in yearling loggerhead sea turtles (Caretta caretta) were determined. Eight juvenile loggerhead sea turtles weighing 1.25 (+/- 0.18) kg were divided into two groups. Four animals received 30 mg/kg of florfenicol i.v., and four received the same dose i.m. Plasma florfenicol concentrations were analyzed by reverse-phase high performance liquid chromatography. After the i.v. dose, there was a biphasic decline in plasma florfenicol concentration. The initial steep phase from 3 min to 1 hr had a half-life of 3 min, and there was a longer slow phase of elimination, with a half-life that ranged from 2 to 7.8 hr among turtles. The volume of distribution varied greatly and ranged from 10.46 to -60 L/kg. Clearance after the i.v. dose was 3.6-6.3 L/kg/hr. After the i.m. injection, there was a peak within 30 min of 1.4-5.6 microg/ml, and florfenicol was thereafter eliminated with a half-life of 3.2-4.3 hr. With either route, florfenicol plasma concentrations were below the minimum inhibitory concentrations for sensitive bacteria within 1 hr. Florfenicol does not appear to be a practical antibiotic in sea turtles when administered at these doses.     

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.     

Endoscopic evaluation of the esophagus and stomach in three loggerhead sea turtles (Caretta caretta) and a Malaysian giant turtle (Orlitia borneensis).

Three loggerhead sea turtles (Caretta caretta) and a Malaysian giant turtle (Orlitia borneensis) were presented with suspected or confirmed esophageal foreign bodies. Esophagoscopy was performed on all turtles, and gastroscopy was performed on three turtles. In all cases, endoscopy was easy to perform, and allowed visualization of most upper gastrointestinal features. The papillated esophagus was easy to navigate, but mucosal papillae in the loggerhead sea turtles prevented examination of the underlying mucosa. The stomach was easily entered and examined in both species, but the working endoscope length (100 cm) prevented inspection of the pyloric antrum and the duodenum in all turtles. The turtles in this report may serve as references for future endoscopic examinations of these species.     

Two herpesviruses associated with disease in wild Atlantic loggerhead sea turtles (Caretta caretta)

Herpesviruses are associated with lung-eye-trachea disease and gray patch disease in maricultured green turtles (Chelonia mydas) and with fibropapillomatosis in wild sea turtles of several species. With the exception fibropapillomatosis, no other diseases of wild sea turtles of any species have been associated with herpesviral infection. In the present study, six necropsied Atlantic loggerhead sea turtles (Caretta caretta) had gross and histological evidence of viral infection, including oral, respiratory, cutaneous, and genital lesions characterized by necrosis, ulceration, syncytial cell formation, and intranuclear inclusion bodies. Nested polymerase chain reaction targeting a conserved region of the herpesvirus DNA-dependent-DNA polymerase gene yielded two unique herpesviral sequences referred to as loggerhead genital-respiratory herpesvirus and loggerhead orocutaneous herpesvirus. Phylogenetic analyses indicate that these viruses are related to and are monophyletic with other chelonian herpesviruses within the subfamily alpha-herpesvirinae. We propose the genus Chelonivirus for this monophyletic group of chelonian herpesviruses.     

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.