Evaluating the Clinical and Cardiopulmonary Effects of Clove Oil and Propofol in Tiger Salamanders (Ambystoma tigrinum)

Our understanding of clinical anesthesia for amphibians is limited. This study represents the first attempt to evaluate the effectiveness of clove oil and propofol as anesthetic agents for tiger salamanders (Ambystoma tigrinum). Twelve apparently healthy adult tiger salamanders were anesthetized in a water bath containing clove oil (450 mg/L of water). After a 2-week wash-out period, 11 of the salamanders were used to evaluate the effectiveness of propofol as an anesthetic agent. Propofol was administered intracoelomically at a dose of 25 mg/kg (n = 5) or 35 mg/kg (n = 6). Heart and respiratory rates were monitored at 5-, 10-, 15-, 20-, 30-, 40-, 50-, 60-, 70-, 80-, 90-, 100-, 120-, 150-, and 180-minute intervals after exposure to the anesthetics. Righting, escape, corneal, superficial pain, and deep pain reflexes were also monitored at these time intervals and ranked as (1) normal, (2) slow, or (3) absent. Surgical anesthesia was determined to be when all of the reflexes were lost. Clove oil produced a surgical level of anesthesia in 67% (8/12) of the salamanders. Propofol administered at 25 mg/kg produced surgical anesthesia in 40% (2/5) of the salamanders, whereas propofol at 35 mg/kg produced surgical anesthesia in 83% (5/6) of the animals. Clove oil did not significantly (P > 0.05) affect respiratory rate at any time, but did decrease heart rate significantly (P < 0.05) after 30 minutes. Propofol produced a significant (P < 0.05) reduction in the respiratory rate at both doses. Heart rate was also found to decrease significantly (P < 0.05) for propofol at 25 mg/kg after 90 minutes and for propofol at 35 mg/kg at 60 minutes and after 80 minutes. Both clove oil and propofol were found to provide a surgical plane of anesthesia for tiger salamanders. However, clove oil provided more rapid onset of the desired level of anesthesia with a longer duration. Although the intracoelomic route for propofol was effective, the time to surgical anesthesia was prolonged. These anesthetics show promise and may prove useful to veterinarians or field biologists working with urodelans.     

Distribution and persistence of atypical porcine pestivirus in experimentally inoculated pigs

Atypical porcine pestivirus (APPV) is a cause of congenital tremors (CTs) in piglets and has been found in swine populations around the globe. Although systemic distribution of the virus has been reported, there is limited information regarding viral localization at the cellular level and distribution at the tissue level. We collected multiple tissues from 2-d-old piglets (n = 36) born to sows inoculated at 45 or 62 d of gestation with APPV via 3 simultaneous routes: intravenous, intranasal, and directly in amniotic vesicles. In addition, 2 boars from APPV-inoculated sows with CT were raised and euthanized when 11 mo old. In situ hybridization performed on tissue samples from piglets demonstrated a broad and systemic distribution of viral RNA including endothelial cells, fibroblasts, and smooth muscle. Labeling in tissues was more pronounced in piglet tissues compared to boars, with the notable exception of diffuse labeling of the cerebellum in boars. Presence of APPV in boar tissues well after resolution of clinical signs suggests persistence of APPV similar to other pestiviruses.     

Study of nebulization delivery of aerosolized fluorescent microspheres to the avian respiratory tract

This study investigated the delivery of an aerosol of monodisperse microspheres to the respiratory tract of birds following aerosol exposure. Adult domestic pigeons (Columbia livia domestica, n = 5 birds per timed treatment) were exposed to an aerosol of fluorescent 1.0 microm diameter carboxylate microspheres for 0.5, 1, 2, or 4 hr. During the aerosolization period, the birds were free-standing in a plexiglass treatment chamber and the aerosol was delivered using a commercial nebulizer. Immediately following aerosol exposure, the birds were euthanatized and the carcasses were intravenously infused with a modified paraformaldehyde/gluteraldehyde fixative. Evaluation of microsphere distribution was performed using a stereoscopic microscope with an epifluorescent module. The results from this study revealed that the amount of aerosolized particles delivered using a commercial nebulizer was proportional to exposure periods. Aerosol exposure periods of 0.5 hr or 1 hr did not result in a readily observable distribution of 1.0 microm fluorescent microspheres to the cranial thoracic, caudal thoracic, or abdominal air sac membranes. This was partly attributed to the relatively low concentration of the individual monodisperse microspheres in the aerosolized suspension. The 2- and 4-hr exposure periods resulted in readily observable deposition of the 1.0 mirom fluorescent microspheres in the cranial thoracic, caudal thoracic, or abdominal air sac membranes, with the 4-hr exposure period resulting in the greatest number of particles on the membrane surfaces. For each of the exposure periods, there was individual animal variation regarding the distribution and relative number of spheres deposited. This study demonstrates the widespread deposition of particles that had an aerodynamic equivalent diameter of approximately 1 microm and provides a better understanding of particle deposition efficiency within the respiratory system following aerosol exposure in birds.     

Characterization of protection from systemic infection and disease by use of a modified-live noncytopathic bovine viral diarrhea virus type 1 vaccine in experimentally infected calves

OBJECTIVE: To evaluate protection against systemic infection and clinical disease provided by use of a modified-live noncytopathic bovine viral diarrhea virus (BVDV) type 1 vaccine in calves challenged with NY-1 BVDV. ANIMALS: 10 calves, 5 to 7 months of age. PROCEDURES: Calves were allocated (n = 5/group) to be nonvaccinated or vaccinated SC on day 0 with BVDV type 1 (WRL strain). Calves in both groups were challenged intranasally with NY-1 BVDV on day 21. Calves' rectal temperatures and clinical signs of disease were recorded daily, total and differential WBC and platelet counts were performed, and serum neutralizing antibody titers against NY-1 BVDV were determined. Histologic examinations and immunohistochemical analyses to detect gross lesions and distribution of viral antigens, respectively, were performed. RESULTS: After challenge exposure to NY-1 BVDV, nonvaccinated calves developed high rectal temperatures, increased respiratory rates, viremia, leukopenia, lymphopenia, and infection of the thymus. Vaccinated calves did not develop high rectal temperatures or clinical signs of respiratory tract disease. Vaccinated calves appeared to be protected against systemic replication of virus in that they did not develop leukopenia, lymphopenia, viremia, or infection of target organs, and infectious virus was not detected in peripheral blood mononuclear cells or the thymus. CONCLUSIONS AND CLINICAL RELEVANCE: The modified-live BVDV vaccine protected calves against systemic infection and disease after experimental challenge exposure with NY-1 BVDV. The vaccine protected calves against infection and viremia and prevented infection of target lymphoid cells.     

Concurrence between clinical and pathologic diagnoses in a veterinary medical teaching hospital: 623 cases (1989 and 1999)

OBJECTIVE: To determine whether there was a decline in the percentage of dogs undergoing necropsies and whether there was substantial agreement or disagreement between clinical and pathologic diagnoses. DESIGN: Retrospective study. ANIMALS: 623 dogs. PROCEDURE: Medical records of hospitalized dogs that died or were euthanatized and necropsied at a veterinary teaching hospital in 1989 and 1999 were reviewed. Clinical and pathologic diagnoses were recorded and compared. RESULTS: There was a significant decline in the necropsy rate of hospitalized dogs that died or were euthanatized in 1999, compared with 1989. In both 1989 and 1999, there was disagreement between the clinical and pathologic diagnoses in approximately a third of the cases. CONCLUSIONS AND CLINICAL RELEVANCE: Despite improved diagnostic methods, the accuracy of diagnosis did not improve significantly in 1999, compared with 1989. Necropsy is the best method to assess overall diagnostic accuracy. Increased availability of teaching funds may promote efforts to have necropsies performed in veterinary teaching hospitals.     

Pharmacokinetics of enrofloxacin after single-dose oral and intravenous administration in the American alligator (Alligator mississippiensis).

The pharmacokinetics of enrofloxacin administered orally and i.v. to American alligators (Alligator mississippiensis) at 5 mg/kg was determined. Plasma levels of enrofloxacin and its metabolite ciprofloxacin were measured using high-performance liquid chromatography and the resulting concentration versus time curve analyzed using compartmental modeling techniques for the i.v. data and noncompartmental modeling techniques for the oral data. A two-compartment model best represented the i.v. data. Intravenous administration of enrofloxacin resulted in an extrapolated mean plasma concentration of 4.19 +/- 4.23 microg/ml at time zero, with average plasma drug levels remaining above 1.0 microg/ml for an average of 36 hr. Plasma volume of distribution for i.v. enrofloxacin was 1.88 +/- 0.96 L/kg, with a harmonic mean elimination half-life of 21.05 hr and mean total body clearance rate of 0.047 +/- 0.021 L/hr/kg. Plasma levels of p.o. enrofloxacin remained below 1.0 microg/ml in all test animals, and average concentrations ranged from 0.08 to 0.50 microg/ml throughout the sampling period. Oral administration of enrofloxacin achieved a mean maximum plasma concentration of 0.50 +/- 0.27 microg/ml at 55 +/- 29 hr after administration, with a harmonic mean terminal elimination half-life of 77.73 hr. Minimal levels of ciprofloxacin were detected after both oral and i.v. enrofloxacin administration, with concentrations below minimum inhibitory concentrations for most susceptible organisms. On the basis of the results of this study, enrofloxacin administered to American alligators at 5 mg/kg i.v. q 36 hr is expected to maintain plasma concentrations that approximate the minimum inhibitory concentration for susceptible organisms (0.5 microg/ml). Enrofloxacin administered to American alligators at 5 mg/kg p.o. is not expected to achieve minimum inhibitory values for susceptible organisms.     

Computer-assisted image analysis of neovascularization in thyroid neoplasms from dogs

OBJECTIVE: To develop a computer-assisted image analysis procedure for quantitation of neovascularization in formalin-fixed paraffin-embedded specimens of thyroid gland tissue from dogs with and without thyroid gland neoplasia. SAMPLE POPULATION: 47 thyroid gland carcinomas, 8 thyroid gland adenomas, and 8 specimens of thyroid tissue from dogs without thyroid gland abnormalities (normal). PROCEDURE: Serial tissue sections were prepared and stained with antibodies against human CD31 or factor VIII-related antigen (factor VIII-rag). The areas of highest vascularity were identified in CD31-stained sections, and corresponding areas were then identified in factor VIII-rag-stained sections. Image analysis was used to calculate the total vascular density in each section, and neovascularization, expressed as a percentage, was determined as the absolute value of the total vascular density derived from factor VIII-rag-stained sections minus the vascular density derived from CD31-stained sections. RESULTS: Mean vascular density of thyroid gland carcinomas derived from CD31-stained sections was significantly greater than density derived from factor VII I-rag-stained sections. This incremental difference was presumed to represent degree of neovascularization. However, significant differences were not detected between vascular densities derived from CD31 and factor VIII-rag-stained sections for either normal thyroid gland tissue or thyroid gland adenomas. No significant correlations were found between vascular density in thyroid gland carcinomas and survival time following surgery. CONCLUSION AND CLINICAL RELEVANCE: A computer-assisted image analysis method was developed for quantifying neovascularization in thyroid gland tumors of dogs. This method may allow identification of dogs with tumors that are most likely to respond to treatment with novel antiangiogenesis agents.     

Detection of parathyroid hormone-related protein in cats with humoral hypercalcemia of malignancy

Background — Increased serum parathyroid hormone‐related peptide (PTHrP) concentration is used to diagnose humoral hypercalcemia of malignancy (HHM) in humans and animals. A commercially available assay for human PTHrP has diagnostic utility in the dog, but has not been assessed in cats. Objectives — The goals of this study were to determine serum or plasma levels of PTHrP in a population of hypercalcemic cats and to determine whether increased PTHrP concentration was associated with malignancy. In addition, we validated immunoradiometric assays (IRMAs) for intact parathormone (iPTH) and PTHrP for use with feline samples. Methods — A retrospective analysis of iPTH and PTHrP results from 322 hypercalcemic cats (ionized calcium concentration >1.4 mmol/L) was performed. Immunoassays for human iPTH and PTHrP (residues 1–84) were validated using standard methods, and reference intervals were calculated using values from 31 healthy adult cats. Hypercalcemic cats were classified as parathyroid‐independent (iPTH <2.3 pmol/L), equivocal (iPTH 2.3–4.6 pmol/L), or parathyroid‐dependent (iPTH >4.6 pmol/L). Seven cats with detectable or increased PTHrP concentrations were evaluated further for underlying disease. Formalin‐fixed neoplastic tissues were immunohistochemically stained using rabbit antibody to human midregion PTHrP. Results — Assays for iPTH and PTHrP showed acceptable precision for feline samples. The reference interval for iPTH was 0.8–4.6 pmol/L and for PTHrP was <1.5 pmol/L. The majority of hypercalcemic cats (263/322, 81.7%) were parathyroid‐independent, with fewer cats in the equivocal (32/322,9.9%) and parathyroid‐dependent (27/322,8.4%) groups. In 31 (9.6%) cats, PTHrP concentration was >1.5 pmol/L (range 1.5–26.6 pmol/L). All 7 cats for which follow‐up information was available had HHM; 6 had carcinomas (4 lung carcinomas, 1 undifferentiated carcinoma, 1 thyroid carcinoma) and 1 had lymphoma. All tumors had mild to moderate positive staining for PTHrP; however, lung carcinomas from normocalcemic cats also stained positive. Conclusions — Human IRMA for PTHrP (1–84) can be used to measure PTHrP in cats. Malignancies, particularly carcinomas, appear to secrete PTHrP and induce HHM in this species. Immunohistochemistry alone cannot predict the occurrence of HHM in cats.