Effect of fluoxetine on blood concentrations of serotonin, cortisol and dehydroepiandrosterone in canine aggression

Canine aggression directed towards people is the most frequent reason for referral to behaviour practices. The serotonergic system and the hypothalamic-pituitary-adrenal (HPA) axis are believed to play an important role in controlling aggression. The selective serotonin reuptake inhibitor fluoxetine is the most commonly used drug in canine aggression. The aim of the present study was to assess the effect of a 30-day-long fluoxetine treatment on the peripheral serotonergic system and the HPA axis in canine aggression. To this end, the concentrations of serum serotonin (5-HT) and plasma cortisol and dehydroepiandrosterone (DHEA) were analysed in a group of aggressive (n = 22) and nonaggressive dogs (n = 9) during pre- (day 0) and posttreatment (day 30) conditions. Treatment caused a significant decrease in 5-HT concentrations (46% in the aggressive group and 32% in the control group). There was a trend towards a rise of DHEA/cortisol ratio values after treatment both in the aggressive and the control group. The determination of blood 5-HT and the DHEA/cortisol ratio could have important clinical applications in the future for deciding which animals might benefit from a given treatment as well as for monitoring the response. Further large-scale studies with this aim should be carried out to obtain sound conclusions.     

Relationship between the serum concentrations of serotonin and lipids and aggression in dogs

The serum concentrations of serotonin and lipids--triglycerides, total cholesterol, low density lipoprotein, high density lipoprotein and very low density lipoprotein--were determined in 18 normal dogs and 23 dogs diagnosed as aggressive on the basis of interviews with their owners and an assessment of their behaviour with reference to a canine overt aggression chart. The serum serotonin levels in the aggressive dogs were significantly lower than in the normal dogs (P<0.01), but the differences in serum lipids between the two groups were not statistically significant.     

Neuro-imaging the serotonin 2A receptor as a valid biomarker for canine behavioural disorders

The serotonergic system is disturbed in different mood and affective disorders, with especially the serotonin (5-HT) 2A receptor involved in impulsive aggressiveness and anxiety. The aim of the study was to evaluate the involvement of the brain 5-HT 2A receptor in dogs with different behavioural disorders. Three groups of drug naive dogs were studied: 22 dogs showing impulsive aggressive behaviour, 22 showing normal behaviour, and 22 showing anxious behaviour. The serotonin 2A receptor was evaluated with Single Photon Emission Computed Tomography (SPECT) and the serotonin 2A receptor-selective radiopharmaceutical (123)I-R91150. A serotonin 2A receptor binding index (BI), proportional to the cortical receptor density, was calculated. A receiver operating characteristic (ROC) analysis was performed to determine cut-off values at which optimal sensitivity and specificity are achieved and to evaluate the general performance of the BI in reflecting the state of the dog, i.e., impulsive aggressive, normal or anxious. Significantly (P<0.0056) altered 5-HT 2A receptor binding indices were found in bilateral frontal, temporal and occipital cortical brain areas of the dogs behaving abnormally, with consistently increased BI in impulsive aggressive dogs and decreased BI in anxious dogs. These results provide clear evidence for a disturbed serotonergic balance in canine impulsive aggression and anxiety disorders. A right frontal cut-off value of ≥1.92 with 86.4% sensitivity and 2.3% (1-specificity) was obtained for the impulsive aggressive dogs. Differentiating the anxious dogs from the rest of the population was possible with a cut-off value of ≤1.73 with 86.4% sensitivity and 18.2% (1-specificity). We conclude that SPECT imaging with the radioligand (123)I-R91150 can be a helpful tool in evaluating the involvement of the serotonin 2A receptor in the complex mechanisms of impulsive aggressive and anxious behaviour. The 5HT-2A binding index of the right frontal cortex appears to be a valid biomarker in differentiating the studied canine behavioural disorders.     

Differences in serotonin serum concentration between aggressive English cocker spaniels and aggressive dogs of other breeds

Aggression is one of the most common behavioral problems in dogs and may have important negative effects on public health, human–animal bond, and animal welfare. There is ample evidence showing a negative correlation between serum serotonin concentration and aggressive behavior in a variety of species, including the domestic dogs. This negative correlation is particularly pronounced in dogs that show impulsive aggression. Data obtained in some previous studies suggest that the English cocker spaniel (ECS) is more likely to show impulsive aggression than other breeds. Therefore, the aim of this study was to analyze possible differences in serum serotonin levels between aggressive ECS and aggressive dogs of other breeds. Nineteen ECSs presented for aggression at the Animal Behavior Service (School of Veterinary Science, Barcelona, Spain) were evaluated and compared with 20 aggressive dogs of other breeds attended at the same center. Serum serotonin levels were measured using an enzyme-linked immunosorbent assay method. Statistical analysis was done using the SPSS 15.0 for Windows. Aggressive ECSs had significantly (P < 0.001) lower levels of serum serotonin than aggressive dogs of other breeds (318.6 ± 67.1 and 852.77 ± 100.58 ng/mL, respectively). Variances were not significantly different between ECSs and other breeds (standard deviation = 449.84 ng/mL vs. 292.47 ng/mL, P > 0.05). This finding may explain why ECSs are more likely to show impulsive aggression than other breeds, and suggests that the ECS could be a good model to study the neurophysiologic mechanisms underlying impulsive aggression.     

Neuro-imaging the serotonin 2A receptor as a valid biomarker for canine behavioural disorders

The serotonergic system is disturbed in different mood and affective disorders, with especially the serotonin (5-HT) 2A receptor involved in impulsive aggressiveness and anxiety. The aim of the study was to evaluate the involvement of the brain 5-HT 2A receptor in dogs with different behavioural disorders.Three groups of drug naive dogs were studied: 22 dogs showing impulsive aggressive behaviour, 22 showing normal behaviour, and 22 showing anxious behaviour. The serotonin 2A receptor was evaluated with Single Photon Emission Computed Tomography (SPECT) and the serotonin 2A receptor-selective radiopharmaceutical ¹²³I-R91150. A serotonin 2A receptor binding index (BI), proportional to the cortical receptor density, was calculated. A receiver operating characteristic (ROC) analysis was performed to determine cut-off values at which optimal sensitivity and specificity are achieved and to evaluate the general performance of the BI in reflecting the state of the dog, i.e., impulsive aggressive, normal or anxious.Significantly (P < 0.0056) altered 5-HT 2A receptor binding indices were found in bilateral frontal, temporal and occipital cortical brain areas of the dogs behaving abnormally, with consistently increased BI in impulsive aggressive dogs and decreased BI in anxious dogs. These results provide clear evidence for a disturbed serotonergic balance in canine impulsive aggression and anxiety disorders. A right frontal cut-off value of ⩾1.92 with 86.4% sensitivity and 2.3% (1-specificity) was obtained for the impulsive aggressive dogs. Differentiating the anxious dogs from the rest of the population was possible with a cut-off value of ⩽1.73 with 86.4% sensitivity and 18.2% (1-specificity).We conclude that SPECT imaging with the radioligand ¹²³I-R91150 can be a helpful tool in evaluating the involvement of the serotonin 2A receptor in the complex mechanisms of impulsive aggressive and anxious behaviour. The 5HT-2A binding index of the right frontal cortex appears to be a valid biomarker in differentiating the studied canine behavioural disorders.     

Aggressive behavior in the English cocker spaniel

A high percentage of aggression problems and a tendency to display noninhibited aggression in the English cocker spaniel (ECS) have been suggested by many authors. The authors of this paper designed a retrospective study to analyze the aggressive behavior of 145 ECSs presented for aggression problems to the Animal Behavior Service at the Barcelona School of Veterinary Medicine's veterinary teaching hospital. Aggressive ECSs were compared with a population of dogs of the same breed presented for a behavior problem other than aggression and with a population of aggressive dogs of other breeds. The most common forms of aggression in the ECS were owner-directed aggression (67.6%), aggression toward unfamiliar people (18.4%), aggression toward unfamiliar dogs (10.1%), and aggression toward family dogs (3.3%).Owner-directed aggression was more common in the ECS than in other breeds, although in similar contexts. In the ECS, the golden coat color was more common in the aggressive dogs than in nonaggressive dogs. ECSs showed impulsive aggression more frequently than aggressive dogs of other breeds. The aim of the study was to analyze cases of aggressive ECSs seen in a referral practice.     

Behavioral characteristics of English Cocker Spaniels with owner-defined aggressive behavior

Information on the behavior of both aggressive and nonaggressive (control) English cocker spaniels (ECS) recruited for genetic analysis is presented. Information from a total of 122 dogs was gathered using questionnaires and owner interviews, resulting in analysis of 52 aggressive (5 aggressive dogs were excluded from the analysis) and 65 control dogs. Owners rated their dogs with 5-point frequency scales for unacceptable behavior toward humans (17 items), fear (20 items), barking (8 items), and aggression toward other dogs (4 items). The results show that serious, repeated growling is the most pronounced single behavior characterizing the aggressive ECS, and this seems to be the most important behavior causing owners to classify their dogs as aggressive. The other main behavior characteristic of the aggressive group was skin-penetrating bites. Snapping and “biting in general” were also frequently reported among these dogs. The objective of this study was to describe some behavioral characteristics of ECS showing owner-defined aggressive behavior, and behavioral differences between these dogs and a group of ECS with excellent behavior.     

Comparison of thyroid analytes in dogs aggressive to familiar people and in non-aggressive dogs

A cross-sectional study was performed in order to examine the association between canine aggression to familiar people and serum concentrations of total thyroxine (TT4), free thyroxine (fT4), thyroxine autoantibodies (T4AA), total triiodothyronine (TT3), free triiodothyronine (fT3), triiodothyronine autoantibodies (T3AA), thyroid stimulating hormone (TSH), and thyroglobulin autoantibodies (TgAA). The subjects were 31 dogs historically aggressive to familiar people and 31 dogs with no history of aggression. Behavioral evaluation and physical examination were completed for each dog in addition to a complete blood count, serum chemistry panel, TT4, fT4 by equilibrium dialysis, TT3, fT3, TgAA, T3AA, and T4AA. Significant differences were found between the two groups with respect to only T4AA, which was increased in the aggressive group, but the concentrations for both groups were within the normal reference range. There were no differences between the two groups in the thyroid analytes most commonly measured by veterinary practitioners evaluating thyroid function in dogs. The results of this study revealed no significant difference between aggressive and non-aggressive dogs in the thyroid concentrations most commonly used to diagnose canine hypothyroidism.     

Analysis of 1040 cases of canine aggression in a referral practice in Spain

A retrospective study was designed to assess the prevalence and clinical presentation of canine aggression problems in a referral practice in Spain. One thousand forty canine aggression cases seen at the Animal Behavior Clinic (Barcelona School of Veterinary Medicine) were analyzed. Each aggression problem was characterized according to 7 predefined diagnostic categories. Breed and sex-related risk factors were calculated by comparing the aggressive dogs to a control population. Aggression problems were the most common behavior complaint (52.28% of all canine behavior cases). Owner-directed aggression was the most frequent of the aggression complaints (38.55%), followed by aggression towards other dogs (34.85%) and unfamiliar people (21.95%). A mean of 1.4 diagnoses of aggression per dog were established. Social conflict aggression towards family members was the most common diagnostic category (35.34%). Sixty nine per cent of aggressive dogs were males and 31% were females, from which only 11 and 12 percent were neutered respectively. Two breeds, the English cocker spaniel and the Catalan sheepdog, presented a particularly high risk for aggression in the studied population.     

Validation of a species-optimized enzyme-linked immunosorbent assay for determination of serum concentrations of insulin in dogs

Background: Measurement of canine serum insulin has relied on methods developed to measure human insulin. A species‐optimized test for measurement of serum insulin in dogs is now commercially available. Objective: The purpose of this study was to validate the canine ELISA for determination of serum insulin concentration in dogs. Methods: Precision was determined by evaluating intra‐ and interassay coefficient of variation (CV), and accuracy was determined by dilution and spike recovery studies. A method comparison study with samples from 34 clinically healthy dogs and 73 dogs examined for various illnesses and disorders (“patients”) was performed using the canine ELISA and an ELISA for human insulin. Biologic relevance of the canine assay was evaluated by measuring insulin in samples collected from 8 healthy dogs after administration of glucagon. A stability study was preformed with 6 samples stored at 20°C, 4–8°C, and ?20°C. Results: For the canine ELISA, intra‐ and interassay CVs were 4.3–7.8% and 4.4–7.7%, respectively. Mean recovery after dilution was 99% and recovery after spiking with porcine insulin was 116%. The canine and human ELISAs correlated well (r²=.94 for healthy dogs, r²=.88 for patient samples). After glucagon injection serum insulin concentrations increased significantly in 8 dogs. Insulin was stable for 30 days in 6 serum samples stored at ?20°C and in most samples for 8 days at 4–8°C. Insulin was stable for <3 days at room temperature (20°C). Conclusions: The new canine serum insulin ELISA had good precision and accuracy and correlated well with the previously used assay.     

Zinc metabolism--a factor in canine aggression?

In order to test the hypothesis of zinc-deficiency as a factor in canine aggression, we examined sera of dangerously aggressive dogs and of behaviourally normal (non-aggressive) dogs for their zinc-contents. The results showed distinctly higher zinc-concentrations (mean +/- SD) in aggressive dogs (1.69 +/- 0.49 micrograms/ml) than in normal non aggressive dogs (0.76 +/- 0.16 microgram/ml).     

Aggressive dogs are characterized by low omega-3 polyunsaturated fatty acid status

Canine aggressive behaviour is one of the most common problems being reported by dog owners. However, the biochemical basis of this phenomenon remains unclear. In humans, alterations in omega-3 plasma polyunsatured fatty acids and elevated omega6/omega-3 ratio have been linked to behavioural alterations, including aggression. Thus far, however, the relationship between plasma polyunsatured fatty acid status and aggression has not been investigated in the dog. In the present study we sought to investigate whether polyunsatured fatty acid status could be altered in plasma of pathologically aggressive Canis familiaris. Eighteen adult male German Shepherd dogs, aged 4.9 ± 0.9 years, showing no clinical signs but aggression, were investigated. Eighteen healthy male dogs, aged 4.8 ± 0.7 years, with a negative history of behavioural and neurological disorders served as controls. Baseline fasting plasma polyunsatured fatty acid composition was determined by gas chromatography. Compared to normal dogs, aggressive dogs showed lower docosahexaenoic acid (22:6 n-3) concentrations and a higher omega6/omega-3 ratio. In addition, they showed reduced cholesterol and bilirubin concentrations compared to their normally behaving counterparts. Altogether, our results suggest that low omega-3 fatty acids may adversely impact behaviour in dogs, resulting in greater propensity to aggression. However, given the cross-sectional design of our study, we cannot claim any causal relationship between the presence of alterations in fatty acid status and canine aggressiveness. Whether omega-3 fatty acids supplementation may be useful to reduce aggressive behaviour in the dog deserves further investigation.     

Increased number of neurons expressing androgen receptor in the basolateral amygdala of pathologically aggressive dogs

The present study aimed to determine the extent to which the androgen receptor (AR) is directly involved in the hormonal modulation of pathological canine aggressive behaviour in the basolateral nuclear group (BNG) of the amygdaloid body. A stereological quantification of AR-positive neurons was performed in the BNGs of normally behaving and aggressive male dogs. The BNG was selected because it is involved in sexual and behavioural activities that are influenced by androgens. In the aggressive dogs the BNG contained a significantly higher number of AR-positive neurons compared with normally behaving dogs suggesting differences in androgen activity within the BNGs of both the groups. However, additional mechanisms are likely to be involved because the AR-negative fraction of BNG neurons was also increased in the aggressive dogs. It was concluded that most of the AR was unliganded because a cytoplasmic staining pattern of AR positivity was observed in the canine BNG neurons. This indicates that genomic androgen actions, which are mediated through the AR are of minor importance in the testosterone modulation of canine aggression within the BNG. Other non-genomic mechanisms through which androgens may exert their action in the BNG are discussed. The aromatase pathway is suggested to be the main mechanism through which testosterone exerts its action within the BNG.     

Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of sarcoptic mange in dogs

Canine scabies is a challenging disease to diagnose because sarcoptic mites are hard to find on skin scrapings. The purpose of this study was to evaluate a serologic enzyme-linked immunosorbent assay (ELISA) as an aid in the diagnosis of canine scabies. In addition, serum samples were obtained post treatment to determine the duration and persistence of circulating scabies antibodies after resolution of natural infection. Nineteen dogs diagnosed with sarcoptic mange and 38 control dogs were tested. Sixteen scabies-infested dogs showed positive pretreatment ELISA results (84.2% sensitivity). Thirty-four control dogs showed negative ELISA results (89.5% specificity). In the 11 scabies dogs from which multiple post treatment serum samples were obtained, detectable antibodies were not present 1 month after treatment in four cases, but were present for 1-4.5 months post treatment in seven dogs. Our results suggest that this scabies ELISA test is useful in the diagnosis of canine scabies.     

Effect of dietary protein content and tryptophan supplementation on dominance aggression, territorial aggression, and hyperactivity in dogs

OBJECTIVE: To evaluate the effect of high- and low-protein diets with or without tryptophan supplementation on behavior of dogs with dominance aggression, territorial aggression, and hyperactivity. DESIGN: Prospective crossover study. ANIMALS: 11 dogs with dominance aggression, 11 dogs with territorial aggression, and 11 dogs with hyperactivity. PROCEDURE: In each group, 4 diets were fed for 1 weeks each in random order with a transition period of not < 3 days between each diet. Two diets had low protein content (approximately 18%), and 2 diets had high protein content (approximately 30%). Two of the diets (1 low-protein and 1 high-protein) were supplemented with tryptophan. Owners scored their dog's behavior daily by use of customized behavioral score sheets. Mean weekly values of 5 behavioral measures and serum concentrations of serotonin and tryptophan were determined at the end of each dietary period. RESULTS: For dominance aggression, behavioral scores were highest in dogs fed unsupplemented high-protein rations. For territorial aggression, [corrected] tryptophan-supplemented low-protein diets were associated with significantly lower behavioral scores than low-protein diets without tryptophan supplements. CONCLUSIONS AND CLINICAL RELEVANCE: For dogs with dominance aggression, the addition of tryptophan to high-protein diets or change to a low-protein diet may reduce aggression. For dogs with territorial aggression, tryptophan supplementation of a low-protein diet may be helpful in reducing aggression.     

Detection of antinuclear antibodies by the Inno‐Lia ANA update test in canine systemic rheumatic disease

Background: Certain systemic autoimmune diseases in dogs are characterized by high titers of circulating antinuclear antibodies (ANA), which can be demonstrated by indirect immunofluorescence (IIF). In an earlier study of IIF‐ANA–positive dogs, the Ouchterlony double immunodiffusion (DID) test was used to identify specific autoantibodies. The DID test has largely been replaced with line blot tests in human diagnostic settings. Objective: The objective of this study was to investigate whether the line blot assay Inno‐Lia ANA update test is a useful tool in demonstrating ANA specificities in canine patients with previously diagnosed IIF‐ANA–positive rheumatic disorders. Methods: Serum samples from 3 clinically healthy control dogs and 20 canine patients with clinical signs of systemic rheumatic disease and documented positive results for IIF‐ANA and DID tests were included in the study. The Inno‐Lia ANA update assay was performed with an anti‐canine detection antibody. Results: Six serum samples that had DID positivity with anti‐spliceosomal small nuclear ribonucleoproteins (snRNP) reactivity showed reactivity to multiple snRNP proteins in the Inno‐Lia test. Samples from 2 dogs that had other types of DID positivity also had clear SmB reactivity and 1 had weak reactivity to RNP‐70K. The other serum samples, including controls, were negative. Conclusions: Using the Inno‐Lia ANA update test, multiple snRNP specificities were demonstrated in some canine patients with autoimmune rheumatic disorders. Other canine autoantibodies may exist that are not detected by this test. Further studies are necessary to characterize the target antigen(s) of these remaining autoantibodies in canine sera.     

Influence of long-term treatment with tetracycline and niacinamide on antibody production in dogs with discoid lupus erythematosus

OBJECTIVE: To evaluate the effect of long-term treatment with tetracycline and niacinamide on antibody production in dogs by measuring postvaccinal serum concentrations of antibodies against canine parvovirus and canine distemper virus. ANIMALS: 10 dogs receiving long-term treatment with tetracycline and niacinamide (treatment group) and 10 healthy dogs (control group). PROCEDURE: The treatment group included 9 dogs with discoid lupus erythematosus and 1 dog with pemphigus foliaceus on long-term treatment (> 12 months) with tetracycline and niacinamide. The control group included 10 healthy dogs with no clinical signs of disease and no administered medications for the past 3 months. Blood samples were obtained from all dogs by jugular venipuncture. Serum antibody titers against canine parvovirus and canine distemper virus antigens were measured, using hemaglutination inhibition and serum neutralization, respectively, and compared between groups. RESULTS: A significant difference in antibody titers between treatment- and control-group dogs was not found. All dogs had protective antibody titers against canine distemper virus, and 8 of 10 dogs from each group had protective titers against canine parvovirus infection. CONCLUSION AND CLINICAL RELEVANCE: These results provide evidence that long-term treatment with tetracycline and niacinamide does not interfere with routine vaccinations and thus does not seem to influence antibody production in dogs.     

Relationships between serum serotonin and serum lipid levels, and aggression in horses

Levels of serum serotonin and serum lipids--triglyceride, total cholesterol, low-density lipoprotein, high-density lipoprotein and very low-density lipoprotein, were determined in normal horses and horses diagnosed with aggression on the basis of a questionnaire survey. Blood serotonin levels in aggressive horses were found to be significantly lower than in non-aggressive horses (P < 0.01), but no association was found with respect to blood lipids.     

Mechanisms of platelet destruction in immune-mediated thrombocytopenia: in vitro studies with canine platelets exposed to heterologous and isologous antiplatelet antibodies

Normal canine platelets coated with heterologous or isologous antiplatelet antibody were interacted with viable canine neutrophils in vitro. Platelet phagocytosis was assessed by detecting nitroblue tetrazolium (NBT) reduction, measuring uptake of opsonised 51Cr-labelled platelets, and electron microscopy. The NBT reduction and uptake of 51Cr-labelled opsonised platelets were markedly increased. Electron microscopy revealed phagocytosis of antibody-coated platelets and their degradation intracellularly. Exposure of canine platelets to rabbit anti-canine platelet antibody in vitro produced morphological changes in platelets and caused serotonin release. Serotonin was not released in the absence of antiplatelet antibody or in the presence of normal rabbit gamma-globulin. Morphological changes in the platelets included disappearance of alpha and dense granules and exaggeration of the open canalicular system. These observations indicate that circulating platelets may be vulnerable to an antiplatelet antibody and that antibody-mediated phagocytosis of platelets is an important mechanism in the pathogenesis of immune-mediated thrombocytopenia.     

Cell-mediated immunity and age at vaccination associated with measles inoculation and protection of dogs against canine distemper.

The antibody-mediated immune response (AMIR) of dogs to measles and canine distemper viruses has been described. However, there is little information on the cell-mediated immune response (CMIR). The AMIR and the CMIR of dogs to canine distemper and to measles were examined. The CMIR was determined for 6 weeks by measuring the 3H-thymidine uptake by immune lymphocytes in the lymphocyte transformation test. Concurrently, canine distemper and measles virus serum-neutralization antibodies were measured by a microtitration serum-neutralization test. Dogs vaccinated with canine distemper virus had a CMIR and an AMIR to canine distemper. However, measles virus-vaccinated dogs had only a CMIR to canine distemper. A CMIR in the absence of an AMIR indicates that cell-mediated immunity is the most important immune mechanism in protecting measles virus-vaccinated dogs against canine distemper. Development of CMIR and AMIR to canine distemper and measles antigens depended on the age of the dog at the time of vaccination. Adult and juvenile dogs had immune responses to both canine distemper and measles. Neither virus, however, elicited an immune response in neonates.