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Objective: To establish pilot data on the plasma concentrations of SC amikacin at 2 doses in red-eared sliders and evaluate concurrent plasma biochemistry parameters., Animals: 8 adult red-eared sliders (Trachemys scripta elegans)., Methods: Amikacin was administered SC at target doses of 5 and 10 mg/kg with a 3-week washout period. Blood samples were collected at 0, 24, 48, 72, and 96 hours postadministration. Plasma amikacin concentrations were quantified using liquid chromatography tandem mass spectrometry. Plasma biochemistry analyses were performed before amikacin administration, 1 week post 5-mg/kg administration, and 1 week post 10-mg/kg administration., Results: Mean maximum amikacin plasma concentrations were recorded 24 hours after 5-mg/kg and 10-mg/kg dosing and were 17.5 ± 2.32 µg/mL and 23.6 ± 2.92 µg/mL, respectively. Mean plasma concentrations after 5-mg/kg dosing steadily decreased to 9.1 ± 0.92 µg/mL by 96 hours postadministration. Amikacin remained detectable in all plasma samples 3 weeks post 5-mg/kg dosing with a mean plasma concentration of 1.04 ± 0.22 µg/mL. Mean plasma concentrations after 10-mg/kg dosing did not decrease over the 96-hour study period. There were no clinically relevant changes in biochemistry parameters., Clinical Relevance: Amikacin persists at detectable plasma levels for at least 3 weeks after SC administration of a 5-mg/kg dose in red-eared sliders, which has not previously been reported in any species. No biochemistry changes consistent with renal toxicity occurred after either dose. Use caution with repeated amikacin dosing in this species until further studies can better characterize cumulative amikacin pharmacokinetics and toxic threshold.

Objective: To evaluate and compare the pharmacokinetic parameters of SC ceftazidime administered at 20 and 40 mg/kg to red-eared sliders., Animals: 8 adult red-eared sliders (Trachemys scripta elegans)., Methods: In a sequential, 2-period study with a 3-week washout period between treatments, ceftazidime was administered SC to turtles at 20 and 40 mg/kg. Blood samples were collected from the subcarapacial sinus at 0, 24, 48, 72, 96, and 120 hours after ceftazidime administration. Plasma ceftazidime concentrations were quantified using reversed-phase HPLC., Results: Mean plasma half-life after 20- and 40-mg/kg dosing was 39.75 ± 8.0 hours and 33.03 ± 6.56 hours, respectively. Mean maximum plasma concentration after 20- and 40-mg/kg dosing was 71.0 ± 15.93 µg/mL and 120.0 ± 30.62 µg/mL, respectively. Mean plasma ceftazidime concentrations remained ≥ 8 µg/mL, the theoretical MIC for various reptile pathogens for all time points., Clinical Relevance: Results indicate that ceftazidime dosed at either 20 or 40 mg/kg produces plasma concentrations exceeding the theoretical MIC of various reptile pathogens for at least 120 hours. An ideal dosing interval could not be determined, as all plasma concentrations remained above the threshold of interest for all time points. Follow-up studies should focus on establishing a dosing interval and more rigorous monitoring for potential adverse effects.

Objective: Determine the hemolytic effect of an 18-µm microaggregate blood filter during in vitro sea turtle whole blood transfusions as well as describe the average diameter of leatherback (Dermochelys coriacea) and Kemp's ridley sea turtle (Lepidochelys kempii) RBCs., Animals: 5 green (Chelonia mydas), 5 loggerhead (Caretta caretta), and 5 Kemp's ridley sea turtles (total n = 15)., Methods: Heparinized sea turtle blood was infused at 60 mL/h through a microbore extension set without and then with a postsyringe, inline 18-µm microaggregate blood filter. Pre- and postfiltration PCV, Hct, total solids, sodium, chloride, potassium, glucose, and free plasma hemoglobin concentrations were measured. With the use of light microscopy and archived blood smears, the maximum and minimum diameter of 20 RBCs from each of the 5 leatherback and 5 Kemp's ridley sea turtles were measured with a calibrated ocular micrometer using 400X magnification., Results: There were no significant differences between pre- and postfiltration samples for Hct, total solids, sodium, chloride, potassium, glucose, and free plasma hemoglobin concentrations; however, there was a significant median postfiltration decrease in PCV of approximately 4%, representing a 13% decrease of the total RBCs transfused. Average maximum diameters for leatherback and Kemp's ridley sea turtle RBCs were 19.7 and 16.1 µm, respectively., Clinical Relevance: Although the 18-µm microaggregate blood filter does not hemolyze transfused sea turtle RBCs and is likely safe for in vivo blood transfusions, the filter's pores may retain a small proportion of infused RBCs given their diameter.

OBJECTIVE To investigate effects of storage duration and temperature on biochemical analytes in plasma from red-eared sliders (Trachemys scripta elegans). ANIMALS 8 red-eared sliders. PROCEDURES Blood samples were collected. Plasma was harvested and analyzed at room temperature (approx 23°C; time = 1 hour) and then fractioned into 0.1-mL aliquots that were stored at room temperature or were refrigerated (4°C) or frozen (-20°C). Biochemical analysis of stored samples was performed at 4 (room temperature), 8 (4°C), 24 (4°C), 48 (4° and -20°C), and 72 (-20°C) hours and at 7 days (-20°C). For each time point for each storage temperature, bias was calculated by subtracting values from the value obtained at 1 hour. Bias was modeled by use of a linear mixed model. RESULTS Storage temperature had a significant effect on several plasma biochemical analytes. In general, aspartate aminotransferase activity and uric acid, total protein, and potassium concentrations increased after storage at 4° and -20°C. Differences in values after storage were mostly within the acceptable range for allowable total error, except for calcium and potassium concentrations for samples stored at -20°C. Both storage temperatures increased variability of measurement results. Results for samples stored at room temperature for 4 hours did not differ significantly from values at 1 hour. Results differed significantly between refrigerated and frozen samples stored for 48 hours. CONCLUSIONS AND CLINICAL RELEVANCE Short-term storage conditions influenced results for some biochemical analytes. These effects should be considered when performing biochemical analyses of plasma samples obtained from red-eared sliders.

OBJECTIVE To determine the pharmacokinetics of meloxicam after single IV and IM injections in red-eared slider turtles (Trachemys scripta elegans). ANIMALS 8 healthy red-eared slider turtles. PROCEDURES Turtles received 1 dose of meloxicam (0.2 mg/kg) IV or IM (4 turtles/route), a 30-day washout period was provided, and then turtles received the same dose by the opposite route. Blood samples were collected at predetermined times for measurement of plasma meloxicam concentration. Pharmacokinetic values for each administration route were determined with a 2-compartment open model approach. RESULTS For IV administration, mean ± SD values of major pharmacokinetic variables were 1.02 ± 0.41 hours for distribution half-life, 9.78 ± 2.23 hours for elimination half-life, 215 ± 32 mL/kg for volume of distribution at steady state, 11.27 ± 1.44 μg•h/mL for area under the plasma concentration versus time curve, and 18.00 ± 2.32 mL/h/kg for total body clearance. For IM administration, mean values were 0.35 ± 0.06 hours for absorption half-life, 0.72 ± 0.06 μg/mL for peak plasma concentration, 1.5 ± 0.0 hours for time to peak concentration, 3.73 ± 2.41 hours for distribution half-life, 13.53 ± 1.95 hours for elimination half-life, 11.33 ± 0.92 μg•h/mL for area under the plasma concentration versus time curve, and 101 ± 6% for bioavailability. No adverse reactions were detected. CONCLUSIONS AND CLINICAL RELEVANCE Long half-life, high bioavailability, and lack of immediate adverse reactions of meloxicam administered IM at 0.2 mg/kg suggested the possibility of safe and effective clinical use in turtles. Additional studies are needed to establish appropriate administration frequency and clinical efficacy.

Objective: To critically evaluate plasma fibrinogen concentration as a diagnostic indicator of inflammation in red-eared sliders (Trachemys scripta elegans)., Design: Prospective induced-disease model and prospective cross-sectional study., Sample: Plasma samples from 12 purpose-bred red-eared sliders and 153 farm-raised red-eared sliders., Procedures: A modification of the Jacobsson method was developed to measure fibrinogen concentration in platelet-poor plasma from red-eared sliders. Purpose-bred turtles had been inoculated with a ranavirus (n = 4) or sterile PBS solution (8) as part of another study. Farm-raised red-eared sliders were categorized as healthy (n = 138) or overtly ill (15) on the basis of physical examination findings at the time of blood sample collection. Samples from 124 of the 138 healthy red-eared sliders were used to establish a fibrinogen concentration reference interval as measured by the modified Jacobsson method. Fibrinogen concentrations in ranavirus-infected and physically ill turtles were compared with those of healthy turtles to determine whether fibrinogen concentration would be a useful diagnostic indicator of inflammation in red-eared sliders., Results: The modified Jacobsson method was reliably used to measure fibrinogen concentration. The fibrinogen concentration reference interval from healthy reproductively active female red-eared sliders was right skewed. Fibrinogen concentration did not differ significantly between healthy red-eared sliders and ranavirus-infected or overtly ill red-eared sliders., Conclusions and Clinical Relevance: A reference interval for red-eared slider plasma fibrinogen concentration was established and partitioned by sex to account for considerable right skewing observed for females. Fibrinogen concentration was not a useful indicator of inflammation in red-eared sliders with ranavirus infection or other overt illnesses.

Objective: To determine the pharmacokinetics of cefovecin sodium after SC administration to Hermann's tortoises (Testudo hermanni)., Animals: 23 healthy adult Hermann's tortoises (15 males and 8 females)., Procedures: Cefovecin (8.0 mg/kg) was injected once in the subcutis of the neck region of Hermann's tortoises, and blood samples were obtained at predetermined time points. Plasma cefovecin concentrations were measured via ultraperformance liquid chromatography coupled to tandem mass spectrometry, and pharmacokinetic parameters were calculated with a noncompartmental model. Plasma protein concentration was quantified, and the percentage of cefovecin bound to protein was estimated with a centrifugation technique., Results: Cefovecin was absorbed rapidly, reaching maximum plasma concentrations between 35 minutes and 2 hours after administration, with the exception of 1 group, in which it was reached after 4 hours. The mean ± SD time to maximum concentration was 1.22 ± 1.14 hours; area under the concentration-time curve was 220.35 ± 36.18 h•μg/mL The mean protein-bound fraction of cefovecin ranged from 41.3% to 47.5%. No adverse effects were observed., Conclusions and Clinical Relevance: Administration of a single dose of cefovecin SC appeared to be well-tolerated in this population of tortoises. Results of pharmacokinetic analysis indicated that the 2-week dosing interval suggested for dogs and cats cannot be considered effective in tortoises; however, further research is needed to determine therapeutic concentrations of the drug and appropriate dose ranges.

Objective-To determine the effect of various UVB radiation sources on plasma 25-hydroxyvitamin D(3) concentrations in Hermann's tortoises (Testudo hermanni). Animals-18 healthy Hermann's tortoises. Procedures-Tortoises were exposed to sunlight in an outdoor enclosure located in the natural geographic range of Hermann's tortoises (n = 6 tortoises) or a self-ballasted mercury-vapor lamp (6) or fluorescent UVB-emitting lamp (6) in an indoor enclosure for 35 days. Plasma samples were obtained from each tortoise on the first (day 0) and last (day 35) days of the study, and concentrations of 25-hydroxyvitamin D(3) were determined. Amount of UVB radiation in enclosures was measured. Results-Mean ± SD plasma 25-hydroxyvitamin D(3) concentrations for tortoises exposed to the mercury-vapor and fluorescent lamps were significantly lower on day 35 (155.69 ± 80.71 nmol/L and 134.42 ± 51.42 nmol/L, respectively) than they were on day 0 (368.02 ± 119.34 nmol/L and 313.69 ± 109.54 nmol/L, respectively). Mean ± SD plasma 25-hydroxyvitamin D(3) concentration for tortoises exposed to sunlight did not differ significantly between days 0 (387.74 ± 114.56 nmol/L) and 35 (411.51 ± 189.75 nmol/L). Mean day 35 plasma 25-hydroxyvitamin D(3) concentration was significantly higher for tortoises exposed to sunlight versus those exposed to mercury-vapor or fluorescent lamps. Sunlight provided significantly more UVB radiation than did the mercury-vapor or fluorescent lamps. Conclusions and Clinical Relevance-Plasma 25-hydroxyvitamin D(3) concentrations differed between tortoises exposed to sunlight and those exposed to artificial UVB sources. Exposure to sunlight at a latitude similar to that of the natural geographic range is recommended for healthy and calcium-deficient tortoises.

Objective: To assess selected clinicopathologic variables at hospital admission (day 1) for cold-stunned Kemp's ridley turtles (Lepidochelys kempii) that died during the first 3 days after admission (nonsurvivors) and turtles that survived (survivors) and to determine the percentage change of each variable from day 1 to day of death (nonsurvivors) or to day 2 or 3 of hospitalization (survivors)., Design: Retrospective case-control study. Animals-64 stranded, cold-stunned Kemp's ridley turtles hospitalized from October 2005 through December 2009., Procedures: Blood gas, pH, Hct, and selected biochemical values in blood samples determined on day 1 and day of death (nonsurvivors; n = 32) or day 2 or 3 of hospitalization (survivors; 32) were obtained from medical records. For each variable, initial values and percentage changes (from initial values to values at the day of death or day 2 or 3 of hospitalization) were compared between survivors and nonsurvivors., Results: Compared with blood analysis findings for survivors, nonsurvivors initially had significantly higher potassium concentration and Pco(2) and significantly lower Po(2), pH, and bicarbonate concentration than did survivors. For the first 2 or 3 days of hospitalization, percentage changes in potassium, lactate, and ionized calcium concentrations were significantly higher and percentage changes in pH and plasma glucose and bicarbonate concentrations were significantly lower in nonsurvivors., Conclusions and Clinical Relevance: At hospital admission, cold-stunned Kemp's ridley turtles were affected by metabolic and respiratory derangements; severe derangements were associated with death. Evaluation of blood gas, pH, Hct, and selected clinicopathologic variables provided useful clinical and prognostic information during rehabilitation of cold-stunned Kemp's ridley turtles.

Objective: To determine reference intervals for concentrations of plasma total protein (TP) and electrophoretogram fractions (ELFs) for healthy, wild loggerhead sea turtles (Caretta caretta) and green turtles (Chelonia mydas) and to assess relationships between TP and ELF concentrations and health status, body size, body mass, and water temperature., Design: Evaluation study., Animals: 437 healthy and 35 ill Atlantic loggerhead sea turtles and 152 healthy and 3 ill Atlantic green turtles., Procedures: Free-ranging turtles were captured from a nuclear power plant intake canal in southern Florida. Plasma samples were obtained from all turtles. Plasma TP and ELF concentrations were measured, and reference intervals were calculated. Wilcoxon rank-sum tests were used to compare TP and ELF values between healthy and ill loggerhead sea turtles. Spearman rank correlations were evaluated between concentrations of TP and ELFs and carapace length, body mass, and water temperature., Results: Reference intervals for TP concentrations were 2.2 to 5.2 g/dL and 2.0 to 5.4 g/dL for loggerhead sea turtles and green turtles, respectively. Except for gamma-globulin, concentrations of ELFs were significantly higher in healthy than in ill loggerhead sea turtles. There was a positive correlation between TP, alpha-globulin, beta-globulin, and gamma-globulin concentrations and water temperature in loggerhead sea turtles and between only TP and alpha-globulin concentrations and water temperature in green turtles., Conclusions and Clinical Relevance: Reference intervals for concentrations of TP and ELFs for healthy, free-ranging loggerhead sea turtles and green turtles can be used in combination with other diagnostic tools to assess health status of sea turtles.

Objective: To document hematologic and plasma biochemical values for a large number of cold-stunned Kemp's ridley turtles at the beginning of rehabilitation, to investigate differences in hematologic and plasma biochemical values of turtles that ultimately survived versus those that died, and to compare values of survivors during convalescence with initial values obtained at the time of admission., Design: Retrospective case series., Animals: 176 stranded, cold-stunned Kemp's ridley turtles hospitalized between 2001 and 2005., Procedures: Hematologic and plasma biochemical values obtained at the time of admission were compared retrospectively for turtles that died versus turtles that survived. Initial results for survivors were compared with convalescent results obtained later in rehabilitation., Results: Turtles that died had significantly greater plasma concentrations of sodium, chloride, potassium, calcium, phosphorus, and uric acid than did turtles that survived. For survivors, values obtained during convalescence for BUN concentration and plasma calcium concentration were significantly greater than initial values obtained at the time of admission, whereas values obtained during convalescence for glucose, sodium, and uric acid concentrations were significantly lower than initial values., Conclusions and Clinical Relevance: Cold-stunned Kemp's ridley turtles may be affected by electrolyte derangements, dehydration, and decreased renal function. Hematologic and plasma biochemical evaluation of such turtles provided useful clinical and prognostic information during the rehabilitation process.

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.

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.

Objective: To identify and characterize blood cells from free-ranging Hawaiian green turtles, Chelonia mydas., Sample Population: 26 green turtles from Puako on the island of Hawaii and Kaneohe Bay on the island of Oahu., Procedure: Blood was examined, using light and electron microscopy and cytochemical stains that included benzidine peroxidase, chloroacetate esterase, alpha naphthyl butyrate esterase, acid phosphatase, Sudan black B, periodic acid-Schiff, and toluidine blue., Results: 6 types of WBC were identified: lymphocytes, monocytes, thrombocytes, heterophils, basophils, and eosinophils (small and large). Morphologic characteristics of mononuclear cells and most granulocytes were similar to those of cells from other reptiles except that green turtles have both large and small eosinophils., Conclusions: Our classification of green turtle blood cells clarifies improper nomenclature reported previously and provides a reference for future hematologic studies in this species.

Paired blood samples were collected from the postoccipital venous plexus and jugular vein of desert tortoises (Gopherus agassizii) for hematologic and plasma biochemical analyses. Comparison of hematologic values revealed significantly (P < or = 0.05) lower PCV, RBC count, WBC count, and hemoglobin values for samples obtained from the occipital site. When comparisons were made between plasma biochemical values for the 2 sites, significant (P < or = 0.05) differences were measured for: glucose, potassium, chloride, uric acid, calcium, phosphorous, total protein, albumin, globulin, alkaline phosphatase, aspartate transaminase, alanine transaminase, and total cholesterol. Significant differences between hematologic and plasma biochemical values from the occipital region samples vs jugular vein samples were attributed to hemodilution of the occipital region samples with extravascular fluid or lymph or both.

We evaluated the effect of anticoagulant (lithium heparin, sodium heparin, or none) and type of autoanalyzer on selected blood biochemical values of the loggerhead sea turtle (Caretta caretta). More differences were observed between the analytes in serum and those in the 2 types of plasma than were observed between the 2 types of plasma. Differences in electrolyte concentrations were not significant when plasma from sodium-heparinized blood was compared with plasma from lithium-heparinized blood. Serum is not recommended for reptilian studies because clot formation is unpredictable and because the time required for clotting may allow substantial changes in the chemical composition of the sample. For most determinants, values varied more between the 2 types of autoanalyzers than among the 3 anticoagulant treatments. These sources of variation must be considered when performing comparative studies.

Morphologic and cytochemical staining characteristics of erythrocytes, leukocytes, and thrombocytes of the desert tortoise (Gopherus agassizii) were evaluated, using blood smears prepared from 23 healthy tortoises of Kern County, Calif. Special emphasis was placed on differentiating features of the various leukocytes and thrombocytes. A variety of cytochemical stains, including benzidine peroxidase, Sudan black B, chloroacetate esterase, alpha-naphthyl butyrate esterase, acid phosphatase, leukocyte alkaline phosphatase, periodic acid-Schiff, and toluidine blue were used. Heterophils had a characteristic, large, focal area of positive staining with chloroacetate esterase, alpha-naphthyl butyrate esterase, and acid phosphatase. Eosinophils stained diffusely positive with benzidine peroxidase, allowing differentiation of this leukocyte from heterophils. Thrombocytes stained focally positive with periodic acid-Schiff, allowing differentiation of these cells from lymphocytes, which stained uniformly negative. An intracytoplasmic body, commonly observed within erythrocytes, was considered ultrastructurally to represent a degenerate organelle.