Lactocin AL705 as quorum sensing inhibitor to control Listeria monocytogenes biofilm formation.

AIMS: The control of Listeria monocytogenes biofilm formation using lactocin AL705 bacteriocin at sub-minimum inhibitory concentrations (MICs) through an antiquorum sensing strategy, was preliminarily investigated. METHODS AND RESULTS: The screening for biofilm formation of different Listeria species at 10°C allowed selecting L. monocytogenes FBUNT for its use as biofilm producer. MIC and minimum bactericidal concentration of lactocin AL705 purified extract against the pathogen was determined. Bacteriocin sub-MICs were used to evaluate biofilm reduction. Concentrations between 2·5-20 AU ml-1 of lactocin AL705 produced significant decreases in biofilm formation without affecting the growth of the pathogen after 3 days of incubation. When bacteriocin concentrations (5-20 arbitrary units per millilitre (AU ml-1 )) were investigated as quorum sensing (QS) inhibitors using Vibrio harveyi as reporter strain, a significant reduction in luminescence by lactocin AL705 (20 AU ml-1 ) was observed. Even when L. monocytogenes produced AI-2 like molecules as recognized by the reporter strain, bacteriocins did not interfere with this compound. CONCLUSION: Antilisterial lactocin AL705 used to disrupt QS through a signal molecule inactivation was able to control L. monocytogenes FBUNT biofilm formation. Other molecule(s) different from the AI-2 involved during biofilm formation could be acting as target of the bacteriocin. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of bacteriocins derived from food-grade micro-organisms as a QS inhibition represents an effective strategy to control pathogens as well as an environmentally friendly sanitation method to mitigate postprocessing food contamination.

Effects of concurrent canine Cushing's syndrome and diabetes Mellitus on insulin requirements, trilostane dose, and survival time.

Trilostane and insulin requirements and survival time of dogs with concurrent naturally-occurring Cushing's syndrome (CS) and diabetes mellitus (DM) has not been fully investigated. This retrospective study evaluated trilostane and insulin doses in dogs with concurrent CS and DM compared to dogs with only CS or DM. Additionally, a survival analysis was performed using a Kaplan-Meier survival curve. Survival time was compared through Log-rank test. Cox proportional regression method was used to screen predictor factors of death in dogs with CS, DM or concurrent CS and DM. A total of 95 dogs were included, 47 dogs had CS, 31 dogs had DM and 17 dogs had concurrent CS and DM. After long-term follow-up, dogs with concurrent CS and DM required higher final median doses of insulin than dogs with DM [0.90 (0.73-1.1) vs 0.67 (0.55-0.73) u/kg/12 h; P = 0,002]. Conversely, the median trilostane requirements in dogs with concurrent CS and DM did not differ from the median trilostane requirements of dogs with CS [1.52 (0.76-2.80) vs 1.64 (1.19-4.95) mg/kg/day; P = 0.283]. No statistical difference was found for the median survival time between dogs with CS and dogs with concurrent CS and DM (1245 vs 892 days; p = 0.152). Although, median survival time of dogs with DM was not reached, it was longer than median survival time of dogs with CS and DM (892 days; P = 0.002). In conclusion, diabetic dogs with concurrent CS need higher insulin doses and have a shorter survival time compared to diabetic dogs without CS.

A mediastinal mass causing Budd-Chiari-like syndrome in a cat.

Background: Budd-Chiari syndrome (BCS) is considered a rare condition in cats that is characterized by the obstruction of the hepatic venous outflow tract from the level of the small hepatic veins to the level of the termination of the inferior vena cava into the right atrium in the absence of cardiac or pericardial disease, or sinusoidal obstruction syndrome. Case description: This report presents a 13-year-old cat with a two-week history of progressive lethargy, inappetence, weight loss, and abdominal distension. Findings/treatment and outcome: The radiological study was consistent with pleural effusion, as well as alveolar and interstitial pulmonary patterns. Ultrasonography confirmed hepatic venin congestion and ascites. Abdominocentesis revealed a modified transudate. A computed tomography (CT) angiography showed a mass at the level of the caudal mediastinum that compressed the caudal vena cava (CVC). Mediastinal lymphoma was considered the most likely differential diagnosis. These findings were interpreted as Budd-Chiari-like syndrome (BCLS) secondary to a mediastinal mass although, unfortunately, no further diagnostic or treatment procedures were accepted by the owners. BCLS is a rare condition in cats, where most of the reported cases occurred as a result of obstruction of the caudal vena cava. In this report, BCLS was caused by a mass located in the caudal mediastinum oppressing the caudal vena cava. Conclusion: This is the first report of BCLS in cats diagnosed by CT angiography, and it shows the value of this technique to define the origin and extent of the mass and to evaluate the presence or absence of metastatic lesions.

Kidney function and glucose metabolism in overweight and obese cats.

Background: In people, obesity and prediabetes mellitus might predispose to chronic kidney disease (CKD).Aims: To assess the association of overweight [Body condition score (BCS) >5] and glucose metabolism alterations, with established or potential markers of CKD. In addition, fructosamine and fasted blood glucose were compared as predictors of early abnormal glucose metabolism.Methods: 54 clinically healthy cats were included in a cross-sectional study comprising 25 neutered males and 29 (28 neutered) females aged 7.2 (5.5-9.4) years. Two potential markers of CKD, namely urinary free active transforming growth factor-ß1-creatinine ratio and urinary retinol binding protein-creatinine ratio were measured along with other parameters to assess CKD. A receiver operating curve was used to identify the best sensitivity and specificity of fructosamine to identify cats with fasting glucose >6.5 mmol/L.Results: No association was found between BCS and markers of CKD. Fructosamine was greater in cats with fasting glucose >6.5 mmol/L compared to those with fasting glucose ≤6.5 mmol/L. A fructosamine concentration ≥250 µmol/L was able to detect cats with hyperglycemia with a sensitivity of 77% and a specificity of 65%. Furthermore, fructosamine was more strongly correlated with fasting glucose than albumin-corrected fructosamine (r = 0.43, p = 0.002 vs r = 0.32, p = 0.026). Cats with higher fructosamine had lower serum symmetric dimethylarginine concentrations.Conclusion: The present study does not suggest an effect of obesity on renal function in domestic cats.Clinical relevance: Fructosamine might be of value for the diagnosis of prediabetes mellitus in cats.

Comparison of non-selective adrenocorticolysis with mitotane or trilostane for the treatment of dogs with pituitary-dependent hyperadrenocorticism.

Forty-six dogs with pituitary-dependent hyperadrenocorticism were treated with mitotane by the non-selective adrenocorticolysis protocol and 40 were treated twice a day with trilostane. The treatment groups were compared by chi-squared tests, and survival data were analysed using Kaplan-Meier survival plots and a Cox proportional hazard method. The non-selective adrenocorticolysis protocol was very effective (89 per cent), its toxicity was moderate (24 per cent) and there were fewer recurrences (29 per cent) than reported with the classical selective adrenocorticolysis protocol (58 per cent). In a multivariate model, age and bodyweight at diagnosis were significantly negatively correlated with survival time. The median survival time of the dogs treated with trilostane twice a day (900 days) was longer (P=0.05) than that of the dogs treated with mitotane (720 days).

Organization of hindlimb nerve projections to the rat spinal cord: a choleragenoid horseradish peroxidase study.

The aim of the present study has been to investigate the projections of hindlimb muscle afferent fibers to the spinal cord with particular emphasis on the ventral horn and the column of Clarke. Following transections of the appropriate ventral roots, injections of the B-subunit of cholera toxin conjugated to horseradish peroxidase were made into the tibial, peroneal, hamstring, superior gluteal, femoral, and obturator nerves in one group of adult rats. In another group of rats, similar experiments were done with intact ventral roots in order to map the location in the ventral horn of the motoneuron cell columns supplying each investigated nerve. An extensive overlap was found for the different nerve projections to Rexed's laminae V-VII. A somatotopic organization of the nerve projections was seen in the lamina IX cell groups of the ventral horn as well as in the column of Clarke, even though an overlap existed. The densest primary afferent projection from each injected nerve was to its homonymous motoneurons. Only a small to moderate overlap between the projections of the tributary branches of the sciatic nerve was found in the ventral horn, whereas the obturator and femoral nerve projections showed more profound overlap. In the column of Clarke, hindlimb nerves innervating distal muscles projected medially, and nerves innervating proximal muscles projected laterally.

Distribution of lumbar dorsal root fibers in the lower thoracic and lumbosacral spinal cord of the rat studied with choleragenoid horseradish peroxidase conjugate.

Spinal cord projections from lumbar dorsal root ganglia (DRGs) were investigated in adult rats following injections of choleragenoid horseradish peroxidase (B-HRP) into each of the six lumbar DRGs. This method is known to label primarily thick fibers. Labeling was found in all laminae except in the outer part of lamina II. Labeled fibers and terminal-like structures were found 8-13 segments rostral and 1-5 segments caudal to the injected DRGs. A somatotopic organization was revealed in laminae III, where the labeling seemed to be organized in mediolateral zones. Some of these protruded as fingerlike processes through segments rostral and caudal to the root entry level. An interdigitating pattern for these fingerlike processes was seen between some DRGs, while an extensive overlap was found between other DRGs. Many zones were found to correspond to the known central projections of peripheral sensory nerves supplied by the injected ganglion. This suggests that the central projection of a DRG is highly related to the projections of the peripheral nerves included in the DRG. The projections to lamina IV were organized in a similar manner as in lamina III, even though the projections showed a higher degree of overlap than in lamina III. No clear somatotopic organization was found in laminae V-IX. Provided that the topographical relationship between central projections of single peripheral nerves and of DRGs correspond to their peripheral projections, the results of this study, together with results of earlier studies suggest that the outlines of dermatomes are highly related to the territories of peripheral nerves included in the dermatomes.

Cytoarchitectonic organization of the spinal cord in the rat: II. The cervical and upper thoracic cord.

A laminar cytoarchitectonic scheme of the cervical and upper thoracic segments of the rat spinal cord is presented in which Rexed's principles for the cat are applied. The material examined in the current investigation consists of 50-80 microns-thick unstained or Nissl-stained sections, and 2 microns-thick plastic-embedded sections stained with paraphenylenediamine. The cytoarchitectonic organization was found to be basically similar to that of the cat. As in our previous study of the cytoarchitectonic organization of the lower thoracic and lumbosacral spinal cord (Molander et al.; J. Comp. Neurol. 230:133-141, '84), the borderlines between the laminae were often found to be ambiguous, suggestive of zones of transition rather than sharp borders. In addition to the laminar scheme, the distribution of certain important cell groups, including the column of Clarke, the central cervical nucleus, the lateral cervical nucleus, the lateral spinal nucleus, and the internal basilar nucleus, is described.

Transganglionic transport and binding of the isolectin B4 from Griffonia simplicifolia I in rat primary sensory neurons.

The isolectin B4 from Griffonia simplicifolia I binds to a subpopulation of rat small-diameter dorsal root ganglion neurons, and to fibres and presumed terminals in laminae I-II of the spinal cord dorsal horn. In the present study we investigated B4 and B4 conjugated to horseradish peroxidase as potential transganglionic tracers of somatic primary afferent neurons after injection into a peripheral nerve. We also tried to identify the specific subpopulation of dorsal root ganglion neurons that bind and ganglion neurons that bind and transport B4. Following injection of B4 or B4-horseradish peroxidase into the sciatic nerve, labelled presumed terminals that reached peak labelling at two days were found exclusively in regions of the spinal cord gray matter known to receive unmyelinated primary afferent fibres. Almost all dorsal root ganglion cells that transported B4-horseradish peroxidase also bound B4. Cell counts showed that 51% of the dorsal root ganglion neurons were B4-positive and cell area measurements that these were all in the small size range. An extensive overlap was found between B4 and fluoride-resistant acid phosphatase (85%), and between B4 and calcitonin gene-related peptide (59%). Seventeen per cent of the B4-positive cells were substance P-immunoreactive and 9% were immunoreactive to somatostatin. Minimal overlap was seen between B4-positive cells and cells positive for RT97 (3%), a selective marker of primary afferent neurons with myelinated axons. All somatostatin-immunoreactive cells and almost all (95%) of the fluoride-resistant acid phosphatase-positive cells were contained within the B4-positive population. This comprised also 58% of the cells immunoreactive to calcitonin gene-related peptide and 42% of those immunoreactive to substance P. The results obtained show that B4 binds to a subpopulation of unmyelinated primary afferent neurons, and that B4 and B4-horseradish peroxidase can be used as selective transganglionic tracers of this specific cell subpopulation.

Effects of nerve growth factor, brain-derived neurotrophic factor and neurotrophin-3 on the laminar distribution of transganglionically fransported choleragenoid in the spinal cord dorsal horn following transection of the sciatic nerve in the adult rat.

Spinal cord projections from transected sciatic nerves treated with different neurotrophins were investigated in the adult rat following injections of choleragenoid into the proximal stump of the injured nerve. Transganglionically transported choleragenoid labelled primary afferent fibres in all spinal cord dorsal horn laminae except the outer part of lamina II (II(o)), which is almost devoid of labelling. Transection of the sciatic nerve, however, resulted in intense transganglionic choleragenoid labelling in lamina II(o) and in lamina I. In this study, the sciatic nerve was transected bilaterally and 4erve growth factor (6 or 24 microg), brain-derived neurotrophic factor (20 microg), neurotrophin-3 (27 microg) or cytochrome C (8 microg; control substance) was applied unilaterally during postoperative survival times of eight, 16 and 32 days. The animals received bilateral injections of choleragenoid into the injured nerve two days before they were killed. The effect of the axotomy and neurotrophin treatment was evaluated by analysing the extent of choleragenoid and substance P immunoreactivity in the somatotopically appropriate spinal cord dorsal horn regions. At eight days' postoperative survival, laminae I and II(o) on the transected, non-treated side showed much more intense choleragenoid-like immunoreactivity compared to the contralateral transected, nerve growth factor-treated (6 and 24 microg) side. A similar situation was also found in cases treated with the higher dose (24 microg) at 16 days but to a lesser degree when the lower (6 microg) dose was used. After 32 days' survival, there was no detectable side difference in the choleragenoid labelling pattern. At 16 days' survival, the mean area of choleragenoid-positive ganglion cell body profiles in the L5 dorsal root ganglion of the transected, non-treated side was significantly smaller than the mean area of the transected, nerve growth factor-treated (24 microg) neurons. An axotomy-induced depletion of substance P-like immunoreactivity was seen from eight days' survival and onwards, whereas on the nerve growth factor-treated side a clearcut substance P depletion was not observed until 32 days. Brain-derived neurotrophic factor, neurotrophin-3 and cytochrome C had no detectable effects on the distribution of choleragenoid labelling or substance P-like immunoreactivity in the dorsal horn following sciatic nerve transection. In conclusion, peripheral nerve injury-induced expansion of primary afferent choleragenoid labelling in the spinal cord dorsal horn is counteracted by treating the axotomized nerve with nerve growth factor.

Early decline and late restoration of spinal cord binding and transganglionic transport of isolectin B4 from Griffonia simplicifolia I after peripheral nerve transection or crush.

Isolectin B4 from Griffonia simplicifolia I (B4) has a high binding affinity to a large population of unmyelinated primary sensory neurons (Wang et al., Neuroscience 62 (1994) 539-551). Using immunohistochemical techniques, binding and transganglionic transport of B4 in the spinal cord was investigated, both at short and long survival times, after sciatic nerve transection and ligation or crush in the adult rat. Nerve transection and ligation resulted in nearly complete disappearance of B4 immunolabelling in the sciatic nerve territory of the superficial dorsal horn after B4 binding, as well as after transganglionic transport of B4 by 2 weeks postinjury. Partial recovery of both B4 binding and B4 transport was found by 8 months, and nearly complete recovery by 16 months, indicating that reappearance of B4 binding is not critically dependent on peripheral reinnervation. Crush injury made by jeweller's forceps resulted in partial depletion of binding and transport by 2 weeks and a nearly complete recovery by 10 weeks. The results show that binding and transganglionic transport of B4 can be used to label dorsal horn connections of unmyelinated primary afferents during the process of regeneration after crush injury. Furthermore, as B4 binding and transport recover at long survival times in the absence of reestablished peripheral connections, the same techniques can be used to study central primary afferent connections at long survival times after nerve transection. Binding and transganglionic transport of B4 offer alternatives to the use of previous techniques such as transganglionic transport of wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) to study central connections of fine primary afferents after injury.

Simultaneous demonstration of central projections of different peripheral nerves by anti-choleragenoid immunoglobulin markers.

This study demonstrates a fluorescent antibody technique where immunoglobulins (IgG) can be used as transganglionic tracers for the simultaneous demonstration of different primary afferent nerve projections to the spinal cord. Antiserum against choleragenoid (CTB) raised in goat or rabbit was mixed with CTB to obtain a solution consisting of IgG bound to CTB. The mixtures of rabbit and goat IgG bound to CTB were subsequently injected into the sciatic or femoral nerves. Following transganglionic transport, goat and rabbit IgG staining were demonstrated simultaneously in different areas of the L4 dorsal root ganglion, in the spinal cord and in the brain stem, using an immunocytochemical technique. When only goat anti-CTB antibodies or only CTB were injected into the sciatic nerve, no goat IgG immunoreactivity could be detected in the spinal cord.

Ultrastructural observations of non-selective effects of ricin treatment (RCA-120) in the rat dorsal root ganglion.

The effects of ricin (RCA-120) on non-injected dorsal root ganglion (DRG) cells, sharing the same DRG as the injected ones, were studied after ricin injections into the tibial nerve and B-HRP injections into the peroneal nerve. Numerous DRG cells containing B-HRP reaction product and exhibiting signs of advanced degeneration were observed. The findings suggest that ricin may be released from dying injected DRG neurons and taken up by adjacent non-injected DRG cells, which subsequently degenerate.

Dose- and time-dependent selective and non-selective effects of ricin (RCA 120) on rat primary sensory neurons.

The distribution of degenerating fibers in the spinal cord was studied in Fink-Heimer-stained sections following treatment of the tibial nerve with ricinus communis agglutinin (RCA 120). The ricin was either injected into the nerve or applied in a capsule on the transected nerve. Short survival times and low doses of ricin resulted in degeneration in somatotopically appropriate parts of the medial dorsal horn. Longer survival times and higher doses resulted in degeneration which progressively expanded into inappropriate areas in the central and lateral parts of the dorsal horn and in deeper laminae regardless of the mode of application. Furthermore, the effect of a ricin injection into the tibial nerve on transganglionic transport of choleragenoid horseradish peroxidase (B-HRP) in the peroneal nerve was studied following a simultaneous or delayed B-HRP injection. A simultaneous ricin and a B-HRP injection resulted in primary afferent HRP labeling in the gray matter, regardless of the dose of ricin. Following a delayed B-HRP injection almost no primary afferent labeling was seen in the gray matter, unless a very low dose of ricin was injected. This study shows that treatment of a peripheral nerve with a high dose of ricin and a long survival time may result in a considerable non-selective degeneration of fibers in the spinal cord. A selective degeneration may, however, be obtained by using lower doses or shorter survival times.

Demonstration of transganglionically transported choleragenoid in rat spinal cord by immunofluorescence cytochemistry.

Injections of the B-subunit of cholera toxin (CTB) were made into the sciatic nerve of the rat. Following a survival of 2-3 days, the fluorescent antibody technique was used to show that CTB can be utilized as a highly sensitive immunocytochemically detectable transganglionic tracer for primary afferent fibers in the spinal cord. CTB-labeled fibers as well as fibers containing calcitonin gene-related peptide- (CGRP-) or substance P-like immunoreactivity were visualized simultaneously by using different fluorochromes. However, no double labeled fibers were found.

Measurement of serum concentrations of free thyroxine, total thyroxine, and total triiodothyronine in cats with hyperthyroidism and cats with nonthyroidal disease.

OBJECTIVE: To determine the usefulness of measuring serum free thyroxine (T4) concentration as a diagnostic test for hyperthyroidism in cats, and to determine the influence of nonthyroidal disease on free T4 concentration in cats without hyperthyroidism. DESIGN: Prospective case series. ANIMALS: 917 cats with untreated hyperthyroidism, 221 cats with nonthyroidal disease, and 172 clinically normal cats. PROCEDURE: Serum free T4, total T4, and total triiodothyronine (T3) concentrations were measured in cats with untreated hyperthyroidism and cats with nonthyroidal disease. Serum total T4 and T3 concentrations were determined by use of radioimmunoassay, and free T4 concentration was measured by use of direct equilibrium dialysis. Reference ranges for hormone concentrations were established on the basis of results from the 172 clinically normal cats. RESULTS: Sensitivity of serum free T4 concentration as a diagnostic test for hyperthyroidism was significantly higher than the test sensitivity of either total T4 or T3 concentration. Of the 221 cats with nonthyroidal disease, 14 had a high free T4 concentration (ie, false-positive result). Therefore, calculated specificity of measuring serum free T4 concentration as a diagnostic test for hyperthyroidism was significantly lower than test specificity of measuring either the total T4 or T3 concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that determination of free T4 concentration is useful in the diagnosis of hyperthyroidism, especially in cats in which hyperthyroidism is suspected but total T4 and T3 concentrations are within reference ranges. However, because some cats with nonthyroidal disease have high serum free T4 concentrations, hyperthyroidism should not be diagnosed solely on the finding of high free T4 concentration.

Measurement of serum total thyroxine, triiodothyronine, free thyroxine, and thyrotropin concentrations for diagnosis of hypothyroidism in dogs.

OBJECTIVE: To determine whether measurement of baseline serum concentrations of total thyroxine (T4), and triiodothyronine (T3), free T4, and thyrotropin (thyroid-stimulating hormone; TSH) would aid in the diagnosis of hypothyroidism in dogs. DESIGN: Prospective case series. ANIMALS: 54 dogs with hypothyroidism, 54 euthyroid dogs with nonthyroidal disease initially suspected to have hypothyroidism, and 150 clinically normal dogs. PROCEDURE: In the 54 dogs with hypothyroidism, diagnosis was established on the basis of clinical signs, results of routine laboratory and TSH stimulation tests, exclusion of concurrent nonthyroidal disease, and a good clinical response to treatment with L-thyroxine. Blood samples were collected from all dogs and were tested for thyroid hormone and TSH concentrations. Reference ranges for hormone concentrations were established on the basis of results for the 150 clinically normal dogs. RESULTS: Of the 54 hypothyroid dogs, 48 (89%) had low total T4 concentrations, 3 had low-normal concentrations, and 3 had high concentrations because of T4 autoantibodies. In contrast, only 10 (18%) euthyroid dogs had low total T4 concentrations. Only 3 of 31 (10%) hypothyroid dogs had low T3 concentrations; 23 had concentrations within the reference range, and 5 had high concentrations because of T3 autoantibodies. Only 3 of 38 euthyroid dogs had low T3 concentrations. Of the hypothyroid dogs, 53 (98%) had low free T4 concentrations and 1 had a low-normal concentration. Only 4 (7%) euthyroid dogs had low free T4 concentrations. Of the hypothyroid dogs, 41 (76%) had high TSH concentrations, and 13 had TSH concentrations within the reference range. Of the euthyroid dogs, only 4 (8%) had high TSH concentrations. Of all single hormone measurements evaluated, measurement of free T4 concentration had the highest sensitivity (0.98), specificity (0.93), and accuracy (0.95) as a test for hypothyroidism; measurement of total T4 concentration had a lower sensitivity (0.89), specificity (0.82), and accuracy (0.85). Compared with measurement of total or free T4 concentration, measurement of TSH concentration had a lower sensitivity (0.76) and accuracy (0.84) but specificity (0.93) equal to that for measurement of free T4 concentration. When T4 (total or free) and TSH concentrations were evaluated together, specificity was higher than when T4 or TSH concentration was evaluated alone. Only 1 euthyroid dog had low T4 (total and free) and high TSH concentrations. CLINICAL IMPLICATIONS: Results indicate that measurement of serum free T4 and TSH concentrations is useful for diagnosis of hypothyroidism in dogs. About a quarter of the dogs with confirmed hypothyroidism, however, will have serum TSH concentrations within reference limits.

Serum total thyroxine, total triiodothyronine, free thyroxine, and thyrotropin concentrations in dogs with nonthyroidal disease.

OBJECTIVE: To determine whether nonthyroidal disease of various causes and severity is associated with abnormalities in baseline serum concentrations of total thyroxine (T4), triiodothyronine (T3), free T4, or thyrotropin (thyroid-stimulating hormone [TSH]) in dogs believed to be euthyroid. DESIGN: Case-control study. ANIMALS: 223 dogs with confirmed nonthyroidal diseases and presumptive normal thyroid function, and 150 clinically normal dogs. PROCEDURE: Serum total T4, total T3, free T4, and TSH concentrations were measured in dogs with confirmed nonthyroidal disease. Reference ranges for hormone concentrations were established on the basis of results from 150 clinically normal dogs. RESULTS: In dogs with nonthyroidal disease, median serum concentrations of total T4, total T3, and free T4 were significantly lower than those in clinically normal dogs. Median serum TSH concentration in sick dogs was significantly greater than that of clinically normal dogs. When stratified by severity of disease (ie, mild, moderate, and severe), dogs with severe disease had low serum concentrations of total T4, total T3, or free T4 more commonly than did dogs with mild disease. In contrast, serum TSH concentrations were more likely to remain within the reference range regardless of severity of disease. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that serum total T4, free T4, and total T3 concentrations may be low (ie, in the hypothyroid range) in dogs with moderate to severe nonthyroidal disease. Serum TSH concentrations are more likely to remain within the reference range in sick dogs.

Evaluation of a low-dose synthetic adrenocorticotropic hormone stimulation test in clinically normal dogs and dogs with naturally developing hyperadrenocorticism.

OBJECTIVE: To determine whether low doses of synthetic ACTH could induce a maximal cortisol response in clinically normal dogs and to compare a low-dose ACTH stimulation protocol to a standard high-dose ACTH stimulation protocol in dogs with hyperadrenocorticism. DESIGN: Cohort study. ANIMALS: 6 clinically normal dogs and 7 dogs with hyperadrenocorticism. PROCEDURE: Each clinically normal dog was given 1 of 3 doses of cosyntropin (1, 5, or 10 micrograms/kg [0.45, 2.3, or 4.5 micrograms/lb] of body weight, i.v.) in random order at 2-week intervals. Samples for determination of plasma cortisol and ACTH concentrations were obtained before and 30, 60, 90, and 120 minutes after ACTH administration. Each dog with hyperadrenocorticism was given 2 doses of cosyntropin (5 micrograms/kg or 250 micrograms/dog) in random order at 2-week intervals. In these dogs, samples for determination of plasma cortisol concentrations were obtained before and 60 minutes after ACTH administration. RESULTS: In the clinically normal dogs, peak cortisol concentration and area under the plasma cortisol response curve did not differ significantly among the 3 doses. However, mean plasma cortisol concentration in dogs given 1 microgram/kg peaked at 60 minutes, whereas dogs given doses of 5 or 10 micrograms/kg had peak cortisol values at 90 minutes. In dogs with hyperadrenocorticism, significant differences were not detected between cortisol concentrations after administration of the low or high dose of cosyntropin. CLINICAL IMPLICATIONS: Administration of cosyntropin at a rate of 5 micrograms/kg resulted in maximal stimulation of the adrenal cortex in clinically normal dogs and dogs with hyperadrenocorticism.

Serum total thyroxine, total triiodothyronine, free thyroxine, and thyrotropin concentrations in epileptic dogs treated with anticonvulsants.

OBJECTIVE: To determine whether administration of phenobarbital, potassium bromide, or both drugs concurrently was associated with abnormalities in baseline serum total thyroxine (T4), triiodothyronine (T3), free T4, or thyrotropin (thyroid-stimulating hormone; TSH) concentrations in epileptic dogs. DESIGN: Prospective case series. ANIMALS: 78 dogs with seizure disorders that did not have any evidence of a thyroid disorder (55 treated with phenobarbital alone, 15 treated with phenobarbital and bromide, and 8 treated with bromide alone) and 150 clinically normal dogs that were not receiving any medication. PROCEDURE: Serum total T4, total T3, free T4, and TSH concentrations, as well as serum concentrations of anticonvulsant drugs, were measured in the 78 dogs with seizure disorders. Reference ranges for hormone concentrations were established on the basis of results from the 150 clinically normal dogs. RESULTS: Total and free T4 concentrations were significantly lower in dogs receiving phenobarbital (alone or with bromide), compared with concentrations in clinically normal dogs. Administration of bromide alone was not associated with low total or free T4 concentration. Total T3 and TSH concentrations did not differ among groups of dogs. CLINICAL IMPLICATIONS: Results indicate that serum total and free T4 concentrations may be low (i.e., in the range typical for dogs with hypothyroidism) in dogs treated with phenobarbital. Serum total T3 and TSH concentrations were not changed significantly in association with phenobarbital administration. Bromide treatment was not associated with any significant change in these serum thyroid hormone concentrations.