Veterinarians use flumequine (FLU) widely but its toxicological effects are still unclear. FLU doses of 53, 200, or 750 mg/kg were administered orally for six weeks to pubertal male rats for evaluation of their toxicity. Weight gain was poorer after seven days of exposure to FLU 750, but relative weights of the brain, adrenal and thyroid glands, and testes were notably higher. Haematological and lipid profile parameters, cardiac markers, and inorganic phosphate significantly increased in the FLU 750 group. Blood glucose, oestradiol and serum concentrations of immunoglobulins G (IgG) and E (IgE) significantly decreased after treatment. The levels of interleukins 10 (IL-10) and 6 (IL-6) fell significantly in the FLU 200 and FLU 750 groups. Cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1) and cyclooxygenase-2 (Cox-2) expression amplified after treatment. Serum levels of free triiodothyronine (fT3) and free thyroxine (fT4) reduced in the FLU 200 and FLU 750 groups without changes in total T3 or T4 level. All doses of FLU significantly depressed concentrations of thyroid-stimulating hormone (TSH) and testosterone. Histopathology of thyroid glands from rats treated with FLU 750 showed degeneration and depletion of thyroid follicular epithelial cells. Expression of 8-hydroxydeoxyguanosine (8-OHdG) was increased in a dose-dependent manner in the brain, but decreased in the testes. Expression of CYP1A1 increased in the adrenal and pituitary glands. The results of this study suggest that the toxicity of FLU in rats is an effect of its disruptive influence on the pituitary-thyroid hormonal system and on the dysfunction of the immune system.

Objective-To evaluate the effects of deracoxib and aspirin on serum concentrations of thyroxine (T4), 3,5,3'-triiodothyronine (T3), free thyroxine (fT4), and thyroid-stimulating hormone (TSH) in healthy dogs. Animals-24 dogs. Procedure-Dogs were allocated to 1 of 3 groups of 8 dogs each. Dogs received the vehicle used for deracoxib tablets (PO, q 8 h; placebo), aspirin (23 to 25 mg/kg, PO, q 8 h), or deracoxib (1.25 to 1.8 mg/kg, PO, q 24 h) and placebo (PO, q 8 h) for 28 days. Measurement of serum concentrations of T4, T3, fT4, and TSH were performed 7 days before treatment (day -7), on days 14 and 28 of treatment, and 14 days after treatment was discontinued. Plasma total protein, albumin, and globulin concentrations were measured on days -7 and 28. Results-Mean serum T4, fT4, and T3 concentrations decreased significantly from baseline on days 14 and 28 of treatment in dogs receiving aspirin, compared with those receiving placebo. Mean plasma total protein, albumin, and globulin concentrations on day 28 decreased significantly in dogs receiving aspirin, compared with those receiving placebo. Fourteen days after administration of aspirin was stopped, differences in hormone concentrations were no longer significant. Differences in serum TSH or the free fraction of T4 were not detected at any time. No significant difference in any of the analytes was detected at any time in dogs treated with deracoxib. Conclusions and Clinical Relevance-Aspirin had substantial suppressive effects on thyroid hormone concentrations in dogs. Treatment with high dosages of aspirin, but not deracoxib, should be discontinued prior to evaluation of thyroid function.

Hypothyroidism is a possible predisposing factor in a number of disorders of companion psittacine birds. We developed and validated a thyroid-stimulating hormone (TSH) response testing protocol for cockatiels (Nymphicus hollandicus), using 0.1 IU of TSH/bird given IM, with blood sample collection at 0 and 6 hours after TSH, and a commercial radioimmunoassay for thyroxine T4). This protocol was used to document a seasonal sex difference in stimulated T4 values-females responded with higher T4 values than those in males in summer- and a stress-induced depression of baseline T4 values was detected in a group of cockatiels with normal TSH response. An experimental model for mature-onset hypothyroidism in cockatiels was created by radiothyroidectomizing cockatiels with 3.7 MBq (100 microCi) of 131I/bird given IV. Induction of the hypothyroid state was confirmed by baseline T4 concentration, TSH response test results, thyroid pertechnetate scintigraphy, and gross and microscopic examinations. Classical signs of hypothyroidism (eg, hypercholesterolemia, obesity, poor feathering) were lacking or mild at 48 days after thyroid ablation.

Prednisone was given orally to 12 dogs daily for 35 days at an anti-inflammatory dosage (1.1 mg/kg of body weight in divided dose, q 12 h) to study its effect on thyroxine (T4) and triiodothyronine (T3) metabolism. Six of these dogs were surgically thyroidectomized (THX-Pred) and maintained in euthyroid status by daily SC injections of T4 to study peripheral metabolism while receiving prednisone; 6 dogs with intact thyroid gland (Pred) were given prednisone; and 6 additional dogs were given gelatin capsule vehicle as a control group (Ctrl). Baseline T4 concentration after 4 weeks of treatment was not significantly different in dogs of the THX-Pred or Pred group (mean +/- SEM, 2.58 +/- 0.28 or 3.38 +/- 0.58 microgram/dl, respectively) vs dogs of the Ctrl group (2.12 +/- 0.30 microgram/dl). A supranormal response of T4 to thyrotropin was observed in dogs of the Pred group, but the T4 response to thyrotropin-releasing hormone was normal. Baseline T3 concentration in dogs of both steroid-treated groups was significantly (P < 0.05) lower after 2 and 4 weeks of prednisone administration vs pretreatment values, but normalized 2 weeks after prednisone was stopped. Free T3 (FT3) and T4 (FT4) fractions and absolute FT3 and FT, concentrations were not altered by prednisone administration. Reverse T3 (rT3) concentration in vehicle-treated Ctrl dogs (26.6 +/- 3.5 ng/dl) was not different from rT3 concentration in dogs of the THX-Pred (25.7 +/- 4.3 ng/dl) and Pred (28.9 +/- 3.8 ng/dl) groups after 4 weeks of medication. These data indicate that daily oral administration of such anti-inflammatory dose of prednisone for 1 month reduces baseline serum T3 concentration, does not alter serum T4 concentration, and enhances thyroidal sensitivity to thyrotropin.

The objectives of this experiment were to determine serum concentrations of triiodothyronine (T3), thyroxine (T4), and free thyroxine (fT4) at rest, following thyroid-stimulating hormone (TSH) administration, and following phenylbutazone administration in healthy horses. This was done to determine which available laboratory test can best be used for diagnosis of hypothyroid conditions in horses. Serum T3, T4, and fT4 concentrations in serum samples obtained before and after TSH stimulation and following phenylbutazone administration for 7 days were determined. Baseline values ranged from 0.21 to 0.80 ng of T3/ml, 6.2 to 25.1 ng of T4/ml, and 0.07 to 0.47 ng of fT3/dl. After 5 IU of TSH was administered IV, serum T3 values increased to 6 times baseline values in 2 hours. Thyroxine values increased to 3 times baseline values at 4 hours and remained high at 6 hours. Free T4 values increased to 4 times baseline values at 4 hours and remained high at 6 hours. Administration of 4.4 mg of phenylbutazone/kg, every 12 hours for 7 days significantly decreased T4 and fT4 values, but did not significantly affect serum T3 concentrations, It was concluded that a TSH stimulation test should be performed when hypothyroidism is suspected. Measurement of serum fT4 concentrations, by the single-stage radioimmunoassay, does not provide any additional information about thyroid gland function over that gained by measuring T4 concentrations. Phenylbutazone given at a dosage of 4.4 mg/kg every 24 hours, for 7 days did significantly decrease resting T4 and fT4 concentrations, but did not significantly affect T3 concentrations in horses.

Concentrations of serum thyroxine (T4) and 3,5,3'-triiodothyronine (T3) were determined after the administration of freshly reconstituted thyrotropin-releasing hormone (TRH), reconstituted TRH that had been previously frozen, or thyrotropin (TSH) to 10 mature dogs (6 Greyhounds and 4 mixed-breed dogs). Thyrotropin-releasing hormone (0.1 mg/kg) or TSH (5 U/dog) was administered IV; venous blood samples were collected before and 6 hours after administration of TRH or TSH. Concentrations of the T4 and T3 were similar (P > 0.05) in serum after administration of freshly reconstituted or previously frozen TRH, indicating that TRH can be frozen at -20 C for at least 1 week without a loss in potency. Concentrations of T4, but not T3, were higher after the administration of TSH than they were after the administration of TRH (P < 0.01). Concentrations of T4 increased at least 3-fold in all 10 dogs given TSH, whereas a 3-fold increase occurred in 7 of 10 dogs given freshly reconstituted or previously frozen TRH. Concentrations of T4 did not double in 1 dog given freshly reconstituted TRH and in 1 dog given previously frozen TRH. Concentrations of T3 doubled in 5 of 10, 2 of 10, and 5 of 10 dogs given TSH, freshly reconstituted TRH, or previously frozen TRH, respectively. Results suggested that concentrations of serum T4 are higher 6 hours after the administration of TSH than after administration of TRH, using dosage regimens of 5 U of TSH/dog or 0.1 mg of TRH/kg. Additionally, results suggested that Greyhounds have lower concentrations of serum T4 than do mixed-breed dogs, but Greyhounds tend to have higher concentrations of serum T3.

Objective-To establish a sensitive test for the detection of autoantibodies against thyroid peroxidase (TPO) in canine serum samples. Sample Population-365 serum samples from dogs with hypothyroidism as determined on the basis of serum concentrations of total and free triiodothyronine (T3), total and free thyroxine (T4), and thyroid-stimulating hormone, of which 195 (53%) had positive results for at least 1 of 3 thyroid autoantibodies (against thyroglobulin Tg, T4, or T3) and serum samples from 28 healthy dogs (control samples). Procedure-TPO was purified from canine thyroid glands by extraction with detergents, ultracentrifugation, and precipitation with ammonium sulfate. Screening for anti-TPO autoantibodies in canine sera was performed by use of an immunoblot assay. Thyroid extract containing TPO was separated electrophoretically, blotted, and probed with canine sera. Alkaline phosphatase-conjugated rabbit anti-dog IgG was used for detection of bound antibodies. Results-TPO bands were observed at 110, 100, and 40 kd. Anti-TPO autoantibodies against the 40-kd fragment were detected in 33 (17%) sera of dogs with positive results for anti-Tg, anti-T4, or anti-T3 autoantibodies but not in sera of hypothyroid dogs without these autoantibodies or in sera of healthy dogs. Conclusions and Clinical Relevance-The immunoblot assay was a sensitive and specific method for the detection of autoantibodies because it also provided information about the antigen. Anti-TPO autoantibodies were clearly detected in a fraction of hypothyroid dogs. The value of anti-TPO autoantibodies for use in early diagnosis of animals with thyroid gland diseases should be evaluated in additional studies.

The present experiments were performed to examine the effects of acetylcholine (ACh) and carbachol (CC) on thyroxine (T4) release and any possible relation between inhibition of T4 relese and sighaling pathway in guinea pig thyroids. The thyroids were incubated in the medium containing the test agents, samples of the medium wer assayed for T4 by EIA kits. ACh and CC inhibited the TSH-stimulated T4 release. These inhibition were reversed by atropine, but not by d-tubocurarine. The ingibitory effects of ACh on T4 release were prevented by M1- and M3-muscarinic antagonists and its inhibition was also slightly reversed by M2- and M4- muscarinic antagonists. R59022, like ACh and CC, also inhibited the TSH-stimulated T4 release. This inhibition was reversed by protein kinase C inhibitor and Ca2+ channel blocker. The present study suggests that cholinergic inhibition of T4 release from thyroids can be induced mainly by ctivation of the M1- or M3-receptors and that it is mediated throught the muscarinic receptor-stimulated protein kinase C activation

Effects of thyroid-stimulating hormone (TSH) and thyrotropin-releasing hormone (TRH) on plasma concentrations of thyroid hormones, and effects of ACTH and dexamethasone on plasma concentrations of cortisol, were studied in adult male ferrets. Thirteen ferrets were randomly assigned to test or control groups of eight and five animals, respectively. Combined (test + control groups) mean basal plasma thyroxine (T4) values were different between the TRH (1.81 +/- 0.41 microgram/dl, mean +/- SD) and TSH (2.69 +/- 0.87 microgram/dl) experiments, which were performed 2 months apart. Plasma T4 values significantly (P < 0.05) increased as early as 2 hours (3.37 +/- 1.10 microgram/dl) and remained high until 6 hours (3.45 +/- 0.86 microgram/dl) after IV injection of 1 IU of TSH/ferret. In contrast, IV injection of 500 microgram of TRH/ferret did not induce a significant increase until 6 hours (2.75 +/- 0.79) after injection, and induced side effects of hyperventilation, salivation, vomiting, and sedation. There was no significant increase in triiodothyronine (T3) values following TSH or TRH administration. Combined mean basal plasma cortisol values were not significantly different between ACTH stimulation (1.29 +/- 0.84 microgram/dl) and dexamethasone suppression test (0.74 +/- 0.56 microgram/dl) experiments. Intravenous injection of 0.5 IU of ACTH/ferret induced a significant increase in plasma cortisol concentrations by 30 minutes (5.26 +/- 1.21 microgram/dl), which persisted until 60 minutes (5.17 +/- 1.99 microgram/dl) after injection. Plasma cortisol values significantly decreased as early as 1 hour (0.41 +/- 0.13 microgram/dl), and had further decreased by 5 hours (0.26 +/- 0.15 microgram/dl) following IV injection of 0.2 mg of dexamethasone/ferret. These results indicate that IV injection of 1 IU of TSH/ferret is preferable to IV injection of 500 microgram of TRH/ferret for thyroid function testing in adult male ferrets. Results of this study also indicated that when TRH or TSH is used for the thyroid-stimulation test in male ferrets, plasma T4 concentrations, instead of T3, should be used as the indicator of thyroid response. Additionally, IV injection of 0.5 IU of ACTH and 0.2 mg of dexamethasone may be used in ferrets for the ACTH stimulation and dexamethasone-suppression tests, respectively.

Basal serum triiodothyronine (T3) and tetraiodothyronine (T4) concentrations have not been established for the llama (Lama glama). In addition, changes in T3 and T4 concentrations in response to thyroid-stimulating hormone (TSH) administration have not been determined, making clinical evaluation of problems referable to thyroid dysfunction difficult. In study 1, basal T3 and T4 concentrations were determined in serum samples collected from 132 clinically healthy llamas. The llamas were allotted to 3 groups: mature intact or neutered males (group I, n = 25), nonpregnant sexually mature females (group II, n = 21), and pregnant females (group III, n = 86). A mean concentration and a 95% confidence interval were computed for each group. An analysis of variance (ANOVA) indicated that a single confidence interval range (0.45 to 4.18, mean = 1.37 ng T3/ml) adequately defined the normal T3 concentrations for all groups. An ANOVA indicated that the T4 concentrations for the female populations (groups II and III) could be combined with a normal confidence interval range of 39 to 204 ng/ml (mean = 88 ng/ml), whereas a separate range (70 to 220 ng/ml, mean = 124 ng/ml) was determined for the male population. An ANOVA indicated that a single confidence interval range (0.0066 to 0.0321, mean = 0.0146) adequately defined the normal T3/T4 ratio for all groups. In study 2, T3 and T4 concentrations were evaluated in 10 healthy llamas immediately preceding and at 2, 4, 6, 8, and 24 hours after the IV administration of 3 IU of TSH/44 kg of body weight. The T3 and T4 concentrations were significantly higher by 2 hours after TSH administration in both groups. Peak T3 and T4 concentrations were observed at 4 and 8 hours, respectively, after TSH administration. When normalized with respect to serum T3 concentrations in samples collected immediately prior to TSH administration, the maximal increase in predicted T3 concentration was 4.06-fold (80% confidence interval range = 2.99- to 5.50-fold) at 4 hours after TSH administration. The maximal increase in predicted normalized T4 concentration was 2.32-fold (80% confidence interval range = 1.76- to 3.05-fold) at 8 hours after TSH administration. The TSH-stimulated increases in T3 and T4 concentrations at 4 hours were clearly distinguishable from values in samples obtained before TSH administration.