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Abstract

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. (Am J Vet Res 2003;64:304–309)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To develop mouse monoclonal and rabbit polyclonal antibodies against immunoglobulin of Argentine boa constrictors and to demonstrate the ability of these reagents to detect antibody responses in boa constrictors by use of an ELISA and western blot analysis.

Animals—Two 3-year-old Argentine boa constrictors.

Procedure—Boa constrictors were immunized with 2,4-dinitrophenylated bovine serum albumin (DNP-BSA). Each snake received biweekly inoculations of 250 µg of DNP-BSA (half SC, half IP) for a total of 6 inoculations followed by monthly inoculations for 3 months. Preimmune blood samples were collected. Subsequently, blood was collected immediately prior to each booster inoculation. Anti-DNP antibodies were isolated from immune plasma samples by affinity chromatography. Affinity-purified boa anti-DNP immunoglobulin was used for production of polyclonal and monoclonal antibodies. An ELISA and western blot analysis were used to monitor immune responses, for purification of boa anti-DNP immunoglobulin, and for assessment of polyclonal and monoclonal antibody specificity.

Results—A 6-fold increase in optical density (OD405) of immune boa plasma, compared with preimmune plasma, was detected by the polyclonal antibody, and a 12- and 15-fold increase was detected by monoclonal antibodies HL1787 and HL1785, respectively, between weeks 4 and 8. Results of western blot analysis confirmed anti-DNP antibody activity in immunized boa plasma and in affinity column eluates. Polyclonal and monoclonal antibodies detected specific anti-DNP antibody responses in immunized boas.

Conclusions and Clinical Relevance—Polyclonal and monoclonal antibodies recognized boa constrictor immunoglobulin. These antibodies may be useful in serologic tests to determine exposure of snakes to pathogens. (Am J Vet Res 2003;64:388–395)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To describe the gross cross-sectional anatomy of green turtles (Chelonia mydas) and evaluate magnetic resonance imaging (MRI) for detection of internal tumors in green turtles with cutaneous fibropapillomatosis.

Design—Prospective study.

Animals—3 dead green turtles, 1 healthy green turtle, and 8 green turtles with cutaneous fibropapillomatosis.

Procedures—Gross cross-sectional anatomy of a dead turtle was described. Each live turtle underwent a complete physical examination, and dorsoventral whole-body survey radiographic views were obtained. Magnetic resonance imaging was performed in dorsal and transverse planes. Radiographs and magnetic resonance images were examined for evidence of internal nodules. Results were compared with necropsy findings in 5 of 8 turtles.

Results—Nodules in the lungs of 2 turtles were detected via radiography, whereas pulmonary nodules were detected in 5 turtles via MRI. No other visceral nodules were detected via radiography; however, masses in the stomach and adjacent to the bladder and kidneys were detected in 1 turtle via MRI. Other extrapulmonary abnormalities observed at necropsy were not detected on MR images.

Conclusions and Clinical Relevance—MRI may be valuable for detection of internal tumors in green turtles with cutaneous fibropapillomatosis. Nodules were more apparent in the lungs than in other organs. Results of MRI may serve as prognostic indicators for sea turtles undergoing assessment, treatment, and rehabilitation. Clinical application may be limited by cost and availability of MRI technology. (J Am Vet Med Assoc 2004;225:1428–1435)

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in Journal of the American Veterinary Medical Association

Abstract

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 γ-globulin, concentrations of ELFs were significantly higher in healthy than in ill loggerhead sea turtles. There was a positive correlation between TP, α-globulin, β-globulin, and γ-globulin concentrations and water temperature in loggerhead sea turtles and between only TP and α-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.

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To identify risk factors that may predispose California sea lions (Zalophus californianus) to development of cutaneous poxvirus nodules during hospitalization in a rehabilitation center.

Design—Retrospective case-control study.

Animals—90 California sea lions admitted to a rehabilitation center.

Procedure—Hospital records of 275 stranded California sea lions admitted to the rehabilitation center between January 1 and December 31, 2002, were reviewed. All California sea lions (n = 18) that developed ≥ 1 cutaneous poxvirus nodule during hospitalization were classified as cases. Seventy-two California sea lions that did not develop poxvirus lesions during hospitalization were randomly selected (control group). The frequencies of various exposure factors prior to admission, at admission, and during hospitalization for cases and control sea lions were compared by use of logistic regression.

Results—California sea lions that had previously been admitted to the rehabilitation center were 43 times as likely to develop poxvirus lesions as sea lions admitted for the first time; those with high band neutrophil counts (> 0.69 × 103 bands/μL) at admission were 20 times less likely to develop poxvirus lesions than sea lions with counts within reference limits.

Conclusions and Clinical Relevance—Results suggest that sea lions with a history of prior hospitalization or band neutrophil counts within reference limits at admission were more likely to develop poxvirus lesions during hospitalization. Sea lions with histories of hospitalization should be kept in quarantine and infection control measures implemented to help prevent disease transmission to attending personnel and other hospitalized animals. (J Am Vet Med Assoc 2005;227:467–473)

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To determine blood cell morphologic characteristics and hematologic and plasma biochemical reference ranges for iguanas housed in a warm indoor and outdoor environment with regular exposure to direct sunlight.

Design—Original study.

Animals—51 clinically normal iguanas (18 males, 25 females, and 8 juveniles) housed in 3 Florida locations.

Procedure—Blood was collected from the coccygeal or ventral abdominal vein. Any samples that had obvious hemolysis or clot formation were not used. Leukocyte counts were determined manually; other hematologic values were obtained by use of a commercially available cell counter. Plasma biochemical values were determined by use of a spectrophotometric chemistry analyzer. Blood smears were stained with Wright-Giemsa and cytochemical stains for morphologic and cytochemical evaluation.

Results—Hematologic ranges were generally higher in this study than previously reported. Thrombocytes were variable in appearance between individuals and sometimes difficult to distinguish from lymphocytes on a Wright-Giemsa preparation. Concentrations of calcium, phosphorus, total protein, globulins, and cholesterol were significantly higher, and the albumin:globulin ratio was significantly lower, in healthy gravid females than in male or nongravid female iguanas. Nongravid females had significantly higher calcium and cholesterol concentrations, compared with males. The calcium:phosphorus ratio was > 1 in all iguanas. Gravid females had a calcium phosphorus product ranging between 210 and 800. Intracytoplasmic inclusions were identified within the erythrocytes of some iguanas.

Conclusions and Clinical Relevance—Hematologic ranges for iguanas in this study are higher than those reported for iguanas. Sex and age of the iguana should be considered when evaluating biochemical values. Healthy ovulating and gravid females may have significantly increased electrolyte and protein concentrations, but maintain a calcium:phosphorus ratio > 1. (J Am Vet Med Assoc 2001;218:915–921)

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in Journal of the American Veterinary Medical Association

Abstract

Objectives—To characterize protein composition of shell scute of desert tortoises and to determine whether detectable differences could be used to identify healthy tortoises from tortoises with certain illnesses.

Animals—20 desert tortoises.

Procedures—Complete postmortem examinations were performed on all tortoises. Plastron scute proteins were solubilized, scute proteins were separated by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and proteins were analyzed, using densitometry. Two-dimensional immobilized pH gradient-PAGE (2D IPG-PAGE) and immunoblot analysis, using polyclonal antisera to chicken-feather β keratin and to alligator-scale β keratin, were conducted on representative samples. The 14-kd proteins were analyzed for amino acid composition.

Results—The SDS-PAGE and densitometry revealed 7 distinct bands, each with a mean relative protein concentration of > 1%, ranging from 8 to 47 kd, and a major protein component of approximately 14 kd that constituted up to 75% of the scute protein. The 2D IPG-PAGE revealed additional distinct 62- and 68- kd protein bands. On immunoblot analysis, the 14-, 32-, and 45-kd proteins reacted with both antisera. The 14-kd proteins had an amino acid composition similar to that of chicken β keratins. There was a substantial difference in the percentage of the major 14- kd proteins from scute of ill tortoises with normal appearing shells, compared with 14-kd proteins of healthy tortoises.

Conclusions and Clinical Relevance—The major protein components of shell scute of desert tortoises have amino acid composition and antigenic features of β keratins. Scute protein composition may be altered in tortoises with certain systemic illnesses. ( Am J Vet Res 2001;62:104–110)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To characterize retroviruses isolated from boid snakes with inclusion body disease (IBD).

Animals—2 boa constrictors with IBD and 1 boa exposed to an affected snake.

Procedure—Snakes were euthanatized, and tissue specimens and blood samples were submitted for virus isolation. Tissue specimens were cultured with or without commercially available viper heart cells and examined by use of transmission electron microscopy (TEM) for evidence of viral replication. Reverse transcriptase activity was determined in sucrose gradient-purified virus. Western blotting was performed, using polyclonal antibodies against 1 of the isolated viruses. Specificity of the rabbit anti-virus antibody was evaluated, using an immunogold-labeling TEM technique.

Results—3 viruses (RV-1, RV-2, and RV-3) were isolated. The isolates were morphologically comparable to members of the Retroviridae family. Reverse transcriptase activity was high in sucrose gradient fractions that were rich in virus. Polyclonal antibody against RV-1 reacted with proteins of similar relative mobility in RV-1 and RV-2. By use of immunogold labeling, this antibody also recognized virions of both RV-1 and RV-2.

Conclusions and Clinical Relevance—A retrovirus was isolated from boid snakes with IBD or exposed to IBD. Western blot analysis of viral proteins indicated that viruses isolated from the different snakes were similar. Whether this virus represents the causative agent of IBD is yet to be determined. The isolation of retroviruses from boid snakes with IBD is an important step in the process of identifying the causative agent of this disease. (Am J Vet Res 2001;62:217–224)

Full access
in American Journal of Veterinary Research