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.     

Evaluation of canine lymphocyte proliferation: comparison of three different colorimetric methods with the 3H-thymidine incorporation assay.

To evaluate canine lymphocyte stimulation the radioactive thymidine incorporation assay is still the method of choice. In order to find a suitable non-radioactive alternative to the standard 3H-thymidine incorporation assay, proliferation of canine peripheral blood lymphocytes (PBL) was measured with three different colorimetric assays, using the two tetrazolium salts MTT and XTT and 5-bromo-deoxyuridine (BrdU). Isolated canine PBL were stimulated with two different mitogens, Concanavalin A (Con A) and Phytohemagglutinin (PHA), using different culture conditions. Applying statistical analysis we found that BrdU and MTT showed a high correlation to the 3H-thymidine incorporation assay, although the BrdU assay proved to be more sensitive than the MTT assay. No significant correlation between the XTT assay and the radioactive method was demonstrated. Consequently, the BrdU assay is the most suitable alternative to the radioactive method.     

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.     

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.     

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.     

Plasma concentrations of enrofloxacin after single-dose oral administration in loggerhead sea turtles (Caretta caretta).

Plasma concentrations and pharmacokinetics of enrofloxacin were determined in 12 loggerhead sea turtles (Caretta caretta) after oral administration. Six turtles in group 1 and group 2 received enrofloxacin at 10 mg/kg and 20 mg/kg of body weight, respectively. Blood was collected from the cervical sinus before administration and at timed intervals up to 168 hr following administration. Plasma concentrations of enrofloxacin were determined using a microbiologic assay. The mean peak plasma concentration (Cmax) was 4.07 microg/ml and 21.30 microg/ml for groups 1 and 2, respectively. Plasma levels were detectable at 168 hr postadministration, with mean values of 0.380 microg/ml for group I and 2.769 microg/ml for group 2. The mean elimination half-life for enrofloxacin was 37.80 hr for group I and 54.42 hr for group 2. These findings indicated that enrofloxacin is absorbed following oral administration in loggerhead sea turtles, and blood levels are maintained up to 168 hr following administration.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

全球海龟资源和保护概况

世界现存海龟仅2科6属7种,都属国际濒危物种。生物界很难找到像海龟—类受到众多世人强烈关注的动物了,国际上亦十分重视海龟的保护工作,不仅专门成立了海龟专家小组(MTSG),制定了许多国际公约,定期出版《海龟通讯》(MTN)等期刊,还成立了一些影响力广泛的国际组织,并经常召开一些与海龟保护相关的国际会议,掀起了全球范围的拯救海龟大行动。本研究概述了海龟资源现状、致危因素、国际保护措施及成果。近年来全球海龟资源总量呈直线下降趋势,海龟面临的主要生存威胁有:渔业副获、直接捕食、沿海经济发展、环境污染与疾病、全球变暖等。最后,简要介绍了海龟重要的经济、科研、观赏、文化和生态价值,并呼吁全人类共同保护海龟。     

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.