Development of a fecal sample collection strategy for extraction and quantification of fecal immunoglobulin A in dogs

OBJECTIVE: To develop a fecal sample collection strategy and quantification method for measurement of fecal IgA concentrations in dogs. SAMPLE POPULATION: Fecal samples from 23 healthy pet dogs of various breeds. PROCEDURES: Immunoglobulin A was extracted from fecal samples. An ELISA for the measurement of fecal IgA concentrations was established and analytically validated. Intraindividual variation of fecal IgA was determined by calculation of coefficients of variation. A sample collection strategy was developed on the basis of results of intraindividual variation of fecal IgA concentrations. A reference range for fecal IgA concentrations was determined. RESULTS: The method for extraction and quantification of fecal IgA was determined to be sufficiently sensitive, reproducible, accurate, and precise. On the basis of the intraindividual variability of our results, the determined fecal sample collection strategy required analysis of a total of 4 fecal samples/dog, with each fecal sample collected on 2 consecutive days with 28 days between sample collection periods (ie, days 1 and 2 followed by days 28 and 29). Reference range values for fecal IgA concentration were 0.22 to 3.24 mg/g of feces. CONCLUSIONS AND CLINICAL RELEVANCE: Methods of fecal IgA extraction and quantification used in our study allow for identification of dogs with consistently low fecal IgA concentrations. Use of these techniques will enable future investigations into possible associations between low fecal IgA concentrations and signs of gastrointestinal disease in dogs.     

Protein-losing enteropathy in dogs is associated with decreased fecal proteolytic activity

Background: Measurement of proteolytic activity in feces is a traditional method for the diagnosis of exocrine pancreatic insufficiency (EPI). A drawback of this method is the occurrence of falsely low results that may lead to a false‐positive diagnosis of EPI. We hypothesized that intestinal loss of serum proteinase inhibitors in protein‐losing enteropathy (PLE) may inhibit fecal proteolytic activity and be a potential source of false low results. Objective: The objective of this study was to determine the effect of PLE on fecal proteolytic activity in dogs. Methods: Fecal proteolytic activity was measured using a radial diffusion casein digestion assay in 12 samples from 4 clinically healthy control dogs and 30 samples from 16 dogs with PLE. Gastrointestinal protein loss was assessed using an ELISA to determine fecal canine α₁‐proteinase inhibitor concentration. The relationship between the concentration of canine α₁‐proteinase inhibitor in the feces and the diameter cleared in the casein digestion assay was determined. The mean clearing diameter was compared between control dogs and dogs with PLE. Results: A significant negative correlation was observed between fecal canine α₁‐proteinase inhibitor concentration and casein clearing diameter (P < .001, Pearson r=—.6317, r 2 =.3999). Mean clearing diameter was significantly lower in dogs with PLE than in control dogs (12.63 vs 16.83 mm, P < .001, two‐tailed Student's t‐test). Conclusion: Increased fecal loss of α₁‐proteinase inhibitor in dogs with PLE is associated with a significant decrease in fecal proteolytic activity and may result in a false positive diagnosis of EPI.     

The effect of thyroid replacement in dogs with suboptimal thyroid function on owner-directed aggression: A randomized,double-blind,placebo-controlled clinical trial

The efficacy of thyroid hormone replacement therapy (THRT) as treatment for owner-directed aggression in client-owned dogs with borderline low thyroid hormone levels was evaluated by means of a 6-week-long, parallel design, double-blind placebo-controlled study. The designation of “borderline hypothyroid” was made if the dog's free normal thyroxine (T4) value was frankly low or in the bottom 20th percentile of the normal range and either total T4, total triiodothyronine (T3), or free T3 was frankly low or in the bottom 30th percentile of the normal range. The presence of thyroid autoantibodies also qualified a dog for enrollment. Owners recorded the number of aggressive episodes directed toward family members on a daily basis for 8 weeks (2-week baseline phase and 6-week study phase). Twenty-nine dogs completed the study; 14 in a treatment group and 15 in a placebo group. The median number of aggressive episodes per day decreased significantly from baseline in both treated and placebo group dogs in weeks 1-2, 3-4, 5-6, and week 6 (treatment, χ² = 24.8, P < 0.001; placebo, χ² = 20.2, P < 0.001), however the median frequency of aggression was significantly lower in the treatment group (1.21 episodes/day) than in the placebo group (1.71 episodes/day) during week 6 of the study (χ² = 4.047, P = 0.044). Three thyroxine-treated dogs had borderline-low thyroid levels on the final day of the study (day 42). When aggression frequency was compared between the treatment and placebo groups after the removal of 3 thyroxine-treated dogs, the treatment group did not have a significantly lower aggression frequency than the placebo group during week 6 (Kruskal–Wallis statistic: χ² = 3.035, n = 26, P = 0.08). The authors discuss the role of thyroid hormones in the regulation of aggression and other cognitive issues and provide rationale for using THRT in dogs exhibiting owner-directed aggression that also have low normal or baseline thyroid hormone levels.     

Effect of tylosin on dogs with suspected tylosin-responsive diarrhea: a placebo-controlled,randomized, double-blinded,prospective clinical trial

Background

The macrolid antibiotic tylosin has been widely used to treat canine chronic diarrhea, although its efficacy is based on anecdotal reports and experimental studies in dogs and not on strong scientific evidence. The term tylosin-responsive diarrhea (TRD) refers to diarrheal disorders responding to tylosin therapy within a few days. In TRD, the stool remains normal as long as tylosin treatment continues, but diarrhea reappears in many dogs within weeks after discontinuation. The aim of our trial was to assess the effect of tylosin on fecal consistency compared with a placebo treatment in dogs with suspected TRD and additionally to establish whether tylosin in dogs with recurrent diarrhea is as effective as empirical studies and anecdotal reports suggest.

Methods

Subjects comprised 71 client-owned dogs that, according to the owners, had previously been treated successfully with tylosin due to recurrent diarrhea of unknown etiology. At the initial examination, where there were no signs of diarrhea, the dogs were randomly assigned in a 2:1 ratio to a tylosin or placebo group. During a two-month follow-up the owners evaluated the fecal consistency according to previously published guidelines. When diarrhea recurred, either tylosin (25 mg/kg q 24 h, 7 days) or placebo treatment was initiated orally. Treatment outcome was evaluated as the mean of fecal consistency scores assigned during the last three days of the treatment period. To test for differences between the tylosin and placebo group in the proportion of responders, Pearson''s Chi-squared test and Fisher''s exact test were applied.

Results

Sixty-one dogs met the selection criteria and were followed for two months. During the follow-up 27 dogs developed diarrhea and either tylosin or placebo treatment was started. The proportion of dogs with normal fecal consistency at the end of treatment was 85% (17/20) in the tylosin group and 29% (2/7) in the placebo group (Pearson''s Chi-squared test p = 0.0049 and Fisher''s exact test two-sided, p = 0.0114).

Conclusions

Our findings indicate that tylosin is effective in treating recurrent diarrhea in dogs. The dose of 25 mg/kg once daily appears sufficient. No changes specific to TRD were detected in the examinations.     

Evaluation of cerebrospinal fluid lactate and plasma lactate concentrations in anesthetized dogs with and without intracranial disease

The objectives of this study were to establish a reference interval for canine cerebrospinal fluid lactate (CSFL) and to compare CSFL and plasma lactate (PL) concentrations in anesthetized dogs with and without intracranial disease. Using a prospective study, canine blood and cerebrospinal fluid were collected for lactate analysis in 11 dogs with intracranial disease after undergoing magnetic resonance imaging (MRI) (Group ID-MRI), in 10 healthy dogs post-MRI (Group H-MRI), and in 39 healthy dogs after induction of anesthesia (Group H-Sx). Dogs were anesthetized for the procedures using different anesthetic protocols. Neurological scores (NS) and sedation scores (SS) were assessed pre-anesthesia in ID-MRI dogs. The CSFL reference interval [90% confidence interval (CI) for lower and upper limits] was 1.1 (1.0 to 1.2) to 2.0 (2.0 to 2.1) mmol/L. Mean ± SD CSFL concentrations were: ID-MRI, 2.1 ± 0.8; H-MRI, 1.6 ± 0.4; and H-Sx, 1.6 ± 0.2 mmol/L. There was a tendency for higher CSFL in dogs in the ID-MRI group than in those in the H-MRI or H-Sx groups (P = 0.12). There was agreement between CSFL and PL in ID-MRI dogs (P = 0.007), but not in dogs in H-MRI (P = 0.5) or H-Sx (P = 0.2). Of the ID-MRI dogs, those with worse NS had higher CSFL (r² = 0.44). The correlation between CSFL and PL in dogs with intracranial disease and between worse NS and higher CSFL warrants further investigation into the use of CSFL and PL for diagnostic and prognostic purposes.     

Aspects of the humoral immune response to Staphylococcus intermedius in dogs with superficial pyoderma,deep pyoderma and anal furunculosis

Bacterial infection (pyoderma) of the canine skin is largely caused by Staphylococcus intermedius and may be a superficial or deep infection. Pyoderma may be a primary, idiopathic disease or secondary to a range of other dermatological disorders. In this study, the serum concentrations of IgG, IgA, antistaphylococcal IgG and antistaphylococcal IgA were measured by ELISA in normal dogs (n = 22), dogs with idiopathic deep pyoderma (n = 22), atopic dermatitis and superficial pyoderma (n = 24), atopic dermatitis without pyoderma (n = 25), flea bite dermatitis with superficial pyoderma (n = 8), pustular demodicosis (n = 8) and German shepherd dogs with anal furunculosis (n = 28). The serum IgG was significantly increased in dogs with atopy and superficial pyoderma (p < 0.001), and lower than normal in dogs with idiopathic deep pyoderma (p < 0.015). The concentration of serum IgA was significantly lower than normal in dogs with atopy uncomplicated by pyoderma (p < 0.015). The concentration of antistaphylococcal IgG in all clinical sera was significantly elevated (p < 0.001) when compared to normal dogs but concentrations of antistaphylococcal IgA were no greater than in normal dogs. Western blotting analysis for determination of the specificity of serum IgG antistaphylococcal antibody revealed that there were nine major epitopes. Discriminant analysis demonstrated that particular combinations of these epitopes were recognised more frequently by sera from dogs in different clinical groups.     

A prospective,randomized, double-blind,placebo-controlled multisite,parallel-group field study in dogs with osteoarthritis conducted in the United States of America evaluating bedinvetmab,a canine anti-nerve growth factor monoclonal antibody

ObjectiveBedinvetmab, a fully canine anti-nerve growth factor monoclonal antibody, was evaluated in dogs for control of osteoarthritis-related pain in a study conducted to support registration in the USA.Study designRandomized, double-blind, placebo-controlled, multicenter, parallel-group study.AnimalsGeneral practice client-owned dogs with osteoarthritis (n = 272).MethodsDogs were block randomized 1:1 to placebo (saline, n = 137) or bedinvetmab (n = 135; 0.5–1.0 mg kg^–1) administered subcutaneously, once monthly. The primary end point, day 28 Canine Brief Pain Inventory (CBPI) treatment success (TS), required pain severity score (PSS; 0–10) decrease ≥1 and pain interference score (PIS; 0–10) decrease ≥ 2. CBPI TS rates [and number needed to treat (NNT)], change in scores [and standardized effect size (ES)], change in quality of life (QoL) and bedinvetmab half-life were calculated.ResultsSignificant (p < 0.05) improvement with bedinvetmab over placebo occurred (days 28, 42, 56, 84) for CBPI TS. Of cases evaluable for day 28 CBPI TS (placebo, n = 131; bedinvetmab, n = 128), success rates were 36.6% and 47.4%, respectively (p = 0.0410) (NNT, 9.3; PSS and PIS ES, 0.3). CBPI TS increased after the second dose in both groups, plateaued for bedinvetmab at day 42 and decreased for placebo beginning day 84. Day 84 NNT (4.3), PSS (0.4) and PIS (0.5) showed continued improvement with monthly dosing. After the first dose, mean (± standard deviation) bedinvetmab half-life was 19.1 (8.3) days. Adverse events were similar between groups and not considered treatment-related. There was a significant effect of bedinvetmab versus placebo on all CBPI components (PIS, PSS, QoL).Conclusions and clinical relevanceThese results corroborated those previously reported and provide further support of safety and effectiveness of bedinvetmab (0.5–1.0 mg kg^–1) administered subcutaneously at monthly intervals to dogs for control of osteoarthritis-related pain.     

A prospective,randomized, blinded,placebo-controlled multisite clinical study of bedinvetmab,a canine monoclonal antibody targeting nerve growth factor,in dogs with osteoarthritis

ObjectiveBedinvetmab is a canine monoclonal antibody targeting nerve growth factor. This study evaluated the efficacy and safety of bedinvetmab for alleviation of pain associated with osteoarthritis in dogs.Study designDouble-blind, randomized, multicentre, placebo-controlled study.AnimalsClient-owned dogs (n = 287) with osteoarthritis.MethodsDogs were randomized (1:1) to subcutaneous injection with placebo (saline, n = 146) or bedinvetmab (0.5–1.0 mg kg^–1, n = 141) administered monthly. After 3 months, 89 bedinvetmab-treated dogs that responded positively based on owner and veterinarian assessments were administered up to six additional doses of bedinvetmab in a single-armed open-label continuation phase. The primary efficacy end point was treatment success based on the owner-assessed canine brief pain inventory (CBPI) on day 28. Treatment success was defined as ≥ 1 reduction in pain severity score (0–10) and ≥ 2 in pain interference score (0–10).ResultsPercentage treatment success was significantly greater in the bedinvetmab group than in the placebo group from day 7 through all assessed time points (p ≤ 0.0025). On day 28, 43.5% of dogs achieved treatment success with bedinvetmab compared with placebo (16.9%) (p = 0.0017). Treatment success continued through days 56 (50.8%) and 84 (48.2%) in the bedinvetmab group and was < 25% in the placebo group at all time points. Sustained efficacy was demonstrated in the continuation phase. Adverse health events occurred at similar frequencies in both groups. They were considered typical for a population of dogs with osteoarthritis and not related to study treatment. Treatment with bedinvetmab demonstrated a significant effect on all three components of CBPI—pain interference, pain severity, quality of life.Conclusions and clinical relevanceThis study demonstrated the effectiveness and safety of bedinvetmab administered monthly for up to 9 months at 0.5–1.0 mg kg^–1 for alleviation of pain associated with canine osteoarthritis.     

Colloid oncotic pressure and total protein changes in horses during anesthesia fluid therapy

It is unknown whether overlapping or sequential use of nonsteroidal anti‐inflammatory (NSAIDs) results in an increased risk for gastrointestinal (GI) ulceration. The purpose of this pilot study was to evaluate the GI effects of various combinations of an injectable NSAID followed by an oral NSAID, a scenario often employed clinically for management of the pre‐ and post‐operative canine patient. Six healthy Walker hounds received four treatment regimens in a randomized, cross‐over design with a 2 week washout period between each treatment week: carprofen (4 mg kg^–1, SQ) followed by placebo (PO, q24 × 4 days); placebo (SQ) followed by deracoxib (3–4 mg kg^–1, PO, q24 × 4 days); carprofen (4 mg kg^–1, SQ) followed by carprofen (4 mg kg^–1, PO, q24 × 4 days); carprofen (4 mg kg^–1, SQ) followed by deracoxib (3–4 mg kg^–1, PO, q24 ×4 days). Weekly bloodwork (CBC, biochemistry panel, fecal evaluation, fecal occult blood) and daily clinical scoring (TPR, vomiting, diarrhea, appetite) were obtained. GI endoscopy was performed on days –2, 1, 2, 5, and 11 days post treatment of each treatment period and lesions scored using a previously reported 6‐point scale. Data was analyzed using a mixed anova for repeated measures. There were no significant differences in clinical or clinicopathologic data between groups. Within the carprofen‐carprofen and carprofen‐deracoxib groups, lesions worsened by Day 5 (1 day after last oral dose) for the fundic and antral regions (p < 0.05). Fundic, antral and lesser curvature lesions improved by Day 5 in the carprofen‐placebo group and lesser curvature lesions improved in the placebo‐deracoxib group (p < 0.05). No significant within‐group differences were noted for the esophagus, cardia or duodenum. The small number of dogs precludes general conclusions about the safety of sequential NSAID use, but these results suggest that a larger scale study is warranted.     

Zinc–carnosine and vitamin E supplementation does not ameliorate gastrointestinal side effects associated with ciclosporin therapy of canine atopic dermatitis

Chelated zinc–carnosine and vitamin E [GastriCalm^® (GCM); Teva Animal Health] is marketed as an anti‐emetic supplement for dogs to assist the repair of damaged stomach and intestinal mucosa. The purpose of this prospective, double‐blinded, placebo‐controlled trial was to determine whether GCM reduced the frequency of vomiting, diarrhoea and appetite changes during initiation of ciclosporin (Atopica^®; Novartis Animal Health) therapy for the treatment of canine atopic dermatitis. Sixty privately owned dogs diagnosed with atopic dermatitis were randomly assigned to GCM (n = 30) or placebo (n = 30) groups. All dogs received ～5 mg/kg ciclosporin (range, 3.5–5.8 mg/kg) once daily. Dogs <13.6 kg received half a tablet of GCM or placebo; dogs ≥13.6 kg received one tablet once daily. GastriCalm^® or placebo was administered 30 min prior to eating, and the ciclosporin was administered 2 h after feeding. Owners recorded episodes of vomiting, diarrhoea and appetite changes. Dogs were examined on days 0 and 14. Forty‐one of 60 dogs (68.3%) had at least one episode of vomiting, diarrhoea or appetite change, leaving nine placebo dogs (30%) and ten GCM dogs (33.3%) free of adverse events (AE). Twenty‐seven of 60 dogs (45%) vomited, and 15 of 60 (25%) had diarrhoea. There was no significant difference in episodes of individual AEs, but the placebo group had a significantly lower total AE score (summation of episodes of appetite change, vomiting and diarrhoea; P = 0.022). Small dogs (<6.82 kg) had significantly fewer total AEs in both treatment groups and tolerated ciclosporin better than larger dogs (P < 0.05).     

Efficacy of two low-dose oral tylosin regimens in controlling the relapse of diarrhea in dogs with tylosin-responsive diarrhea: a prospective,single-blinded,two-arm parallel,clinical field trial

Background

Despite its wide acceptance as a treatment for canine chronic enteropathies, the macrolide antibiotic tylosin lacks official oral dosage recommendations. Not even textbooks share consensus about the dose; daily recommendations vary from 25 to 80 mg/kg and dosing intervals from one to three times daily.The objective of this prospective, single-blinded, two-arm parallel, clinical field trial was to determine whether doses of 5 mg/kg or 15 mg/kg tylosin administered orally once daily for seven days would have a similar effect on fecal consistency in diarrhea relapses to that of a 25 mg/kg dose of tylosin administered once daily for seven days, a dosage that has proved effective in controlling canine tylosin-responsive diarrhea (TRD). A further objective was to compare the efficacy of the 5 mg/kg and 15 mg/kg tylosin dosages. Fifteen client-owned dogs diagnosed with TRD that had responded to a dose of 25 mg/kg tylosin once daily for seven days were enrolled in the study. After a relapse of diarrhea the dogs were allocated into two groups receiving tylosin orally in doses of either 5 mg/kg or 15 mg/kg once daily for seven days. The owners were blinded to the dosage. The elimination of diarrhea was the main criterion in assessing treatment success. The mean fecal consistency score of the last three treatment days for all dosages, including 25 mg/kg, as evaluated by the owners according to a standardized fecal scoring system, served as the primary outcome measures.

Results

All eight dogs responded to the 5 mg/kg dose, and six of seven dogs responded to the 15 mg/kg dose. The mean fecal consistency scores at the 25 mg/kg tylosin dosage were no significantly different from scores at the 5 mg/kg or 15 mg/kg tylosin dosages (P = 0.672, P = 0.345).

Conclusions

Interestingly, 14/15 (93%) of the dogs responding to a dose of 25 mg/kg tylosin once daily for seven days also responded to the lower dosages at diarrhea relapse. The data indicate that a suitable dose of tylosin for treating diarrhea relapse in canine TRD could be as low as 5 mg/kg once daily for seven days.     

P‐48 Treatment of localized lesions of canine atopic dermatitis with tacrolimus ointment: a blinded,randomized, controlled trial

The purpose of this study was to determine whether tacrolimus ointment (Protopic) decreased the severity of localized lesions of canine atopic dermatitis (AD). Twenty dogs with AD were enrolled if they exhibited skin lesions localized to both front metacarpi. Each foot was randomized to be treated either with 0.1% tacrolimus or placebo (vaseline) ointment twice daily for 6 weeks. The nature of treatment for each foot lesion was concealed from the clinician. Before, and every 2 weeks during the study, erythema, lichenification, oozing and excoriations each were graded on a 10‐point scale (maximal total score: 40). The primary outcome measures consisted of the percentage reduction from baseline of lesional scores, and the number of subjects whose scores had decreased by 50% or greater by the end of the study. Intent‐to‐treat analyses were used. At the beginning of the study, lesional scores were not significantly different between treatment groups. After 6 weeks, the percentage reduction from baseline scores was higher for tacrolimus‐treated sites [median: 63% (95% CI: 39–67)] than for placebo‐treated feet [3% (‐2‐13)] (paired t‐test; P < 0.0001). When tacrolimus was applied, lesions decreased by 50% or greater in 15 dogs (75%), while this benchmark was not reached for any placebo‐treated feet (Fisher's exact test; P < 0.0001). Adverse drug events consisted of minor irritation in some dogs treated with tacrolimus. Results of this randomized, controlled trial suggest that the daily application of 0.1% tacrolimus ointment is useful for reducing the severity of localized skin lesions of canine AD. Funding: Self‐funded.     

Sero‐epidemiology of Canine Leptospirosis in Trinidad: Serovars,Implications for Vaccination and Public Health

A sero‐epidemiological study on canine leptospirosis was conducted in house, stray, farm and hunting dogs, as well as in suspect cases of clinical canine leptospirosis. Serum samples were collected from apparently healthy (vaccinated and non‐vaccinated), house dogs. A questionnaire was administered to the owners to elicit information on risk factors for leptospirosis. The microscopic agglutination test was used to screen for leptospirosis using 17 international serovars. Reciprocal titres of between 100 and <800 were considered as evidence of past exposure while reciprocal titres of 800 or greater were classified as suggestive of acute/current infection. Of a total of 419 serum samples tested, 61 (14.6%) were seropositive for Leptospira agglutinins, 23 (5.5%) had mixed infections and 16 (3.8%) had current infection. Amongst 50 suspected cases of clinical leptospirosis, 24 (48.0%) were seropositive and only 13 (26.0%) had current infection compared with 10 (6.3%) and three (1.9%) of 160 apparently healthy house dogs respectively. The difference was statistically significant (P < 0.05; χ²). Twelve (25.5%) of 47 hunting dogs, 10 (20.4%) of 49 farm dogs and five (4.4%) of 113 stray dogs were seropositive (P < 0.05; χ²). Overall, a total of nine serovars were detected with serovars mankarso, icterohaemorrhagiae RGA, autumnalis and copenhageni being involved in 29 (47.5%), 20 (32.8%), 25 (41.0%) and 10 (16.4%) respectively in 61 seropositive dogs (P < 0.05; χ²). Serovar mankarso was most predominant in seropositive apparently healthy dogs, 37.8% (14/37), suspected clinical cases of leptospirosis, 62.5% (15/24) compared with serovar icterohaemorrhagiae with a frequency of 21.6% (8/37) and 50.0% (12/24), the difference being statistically significant (P < 0.05; χ²). Although all vaccines used for prevention of canine leptospirosis in the country contain serovars canicola and icterohaemorrhagiae, serovar mankarso was mostly associated with infection and disease and may be a good candidate for inclusion in the vaccine used locally. The public health risk posed to owners of dogs infected with Leptospira cannot be over‐emphasized considering the zoonotic nature of the disease.     

Factors associated with development of Canine Infectious Respiratory Disease Complex (CIRDC) in dogs in 5 Canadian small animal clinics

This study investigated the association between presence of respiratory pathogens and development of Canine Infectious Respiratory Disease Complex (CIRDC) in dogs in 5 Canadian small animal clinics. In total, 86 dogs were tested using a commercial PCR respiratory panel; 64 dogs were considered as cases and 22 were control dogs matched by veterinary clinic. No control animals (0/22) were positive for canine parainfluenza virus (CPIV), whereas 27/64 (42%) CIRDC cases were positive. Furthermore, 81% of case dogs tested positive for Mycoplasma cynos, compared with 73% of control dogs. Canine respiratory corona virus (CRCoV) was detected in no control dogs compared with 9.4% of clinical dogs. No animals were positive for any influenza virus type A present in the diagnostic panel. Presence of CPIV was associated (P < 0.01) with the occurrence of CIRDC after adjustment for demographic factors and presence of CRCoV (P = 0.09).     

Concentrations of platelet α2-adrenoceptors,lymphocyte muscarinic receptors,and blood monoamines in dogs (Canis familiaris) affected by canine cognitive dysfunction syndrome

Canine cognitive dysfunction syndrome (CDS) is a neurodegenerative disorder of aged dogs characterized by a progressive decline in cognitive function. In humans and laboratory animals, a variety of neurotransmitter abnormalities have been described in patients affected by age-related dementia. Specifically, the regulatory role of the catecholaminergic, serotonergic, and cholinergic systems has been outlined. The aim of the present study was to measure blood monoamine levels, platelet α₂-adrenergic receptors, and lymphocyte muscarinic receptors in healthy adult and aged dogs and in dogs affected by canine cognitive dysfunction. Based on clinical and behavioral examination, 40 dogs were divided into 3 groups: healthy adults (n = 14), aged dogs (n = 17), and aged dogs affected by canine cognitive dysfunction (n = 9). A significant reduction in plasma levels of norepinephrine and dopamine was observed both in aged dogs (0.16 ± 0.02 ng/mL, P < 0.01; 0.11 ± 02 ng/mL, P < 0.01, respectively) and in CDS dogs (0.14 ± 0.03 ng/mL, P < 0.05; 0.10 ± 00.005 ng/mL, P < 0.01, respectively) compared with adults (0.29 ± 0.04 ng/mL and 0.15 ± 0.02 ng/mL, respectively). No significant differences were observed among groups for α₂-adrenergic receptor concentrations. Canine lymphocytes express 2 distinct classes of muscarinic receptors, characterized by high (HA) and low affinity (LA) for [³H]-N-methyl-scopolamine. A significant age-dependent decrease in HA muscarinic receptors was observed. However, no differences were found between aged dogs (87.65 ± 11.08 sites/cell × 10²) and in CDS dogs (90.17 ± 6.75 sites/cell × 10² ) for HA muscarinic receptor concentrations. As far as LA muscarinic receptors are concerned, CDS dogs showed a significant increase (393.48 ± 63 sites/cell × 10²; P < 0.05) with respect to healthy adult dogs (188.84 ± 16.50 sites/cell × 10²). Our results suggest that the reduction in HA muscarinic receptor-binding sites could be representative of the physiological aging process, whereas the increase in lymphocyte LA muscarinic receptor levels could be related to the cognitive decline.     

A topical aqueous calcineurin inhibitor for the treatment of naturally occurring keratoconjunctivitis sicca in dogs

Purpose The purpose of this study was to evaluate the efficacy of an aqueous calcineurin inhibitor, SCY‐641, in the treatment of naturally occurring canine immune‐mediated keratoconjunctivitis sicca (KCS). Methods A randomized, double‐masked, placebo‐controlled clinical study of 56‐day duration was performed in dogs with naturally occurring immune‐mediated KCS assigned to treatment with either topical twice‐daily aqueous calcineurin inhibitor solution (SCY‐641) or artificial tears (placebo) by the study administrator. Clinical examination and Schirmer tear tests (STT) were performed prior to therapy and at days 7, 14, 28, and 56 after initiation of treatment. Results Twenty dogs were enrolled in the study with ten receiving placebo and 10 receiving SCY‐641 in one or both eyes. No adverse effects were noted with any treatment. There were no significant differences in mean STT values in dogs in group either at day 0 (prior to therapy) or after 7 days of treatment. At 14, 28, and 56 days after initiation of treatment, mean STT and increase in STT over baseline in dogs treated with SCY‐641 were significantly higher than in dogs treated with placebo (P < 0.04). Conclusions SCY‐641 was well tolerated by dogs with naturally occurring KCS, and by 14 days after initiating therapy, dogs treated with SCY‐641 had significantly higher STT than placebo‐treated dogs. These preliminary results indicate that topical SCY‐641, in a stable clear aqueous solution, is efficacious in a spontaneous model of KCS and warrants further evaluation as a treatment of immune‐mediated KCS.     

Canine vector-borne disease in travelled dogs in Germany--a retrospective evaluation of laboratory data from the years 2004-2008

When importing dogs from various Mediterranean countries into Western Europe canine vector-borne infections are often considered as a major issue. Several diseases including babesiosis, leishmaniosis, hepatozoonosis, canine heartworm disease or ehrlichiosis can potentially be endemic in this region and pose a potential health risk for travelling dogs. Information on such infections in travelled dogs is scarce and therefore this study has been undertaken to examine the frequency of vector-borne infections in travelled dogs from the years 2004-2008. A total of 997 samples were screened by direct and/or indirect methods. Total seroprevalence was 7.5% with individual seroprevalence for the 3 species Leishmania spp., Ehrlichia canis and Babesia canis spp. ranging from 3.1 to 4.9%. Total detection rate for pathogens by direct methods was 3.5%. Ninteen Giemsa-stained blood smears were positive for large Babesia. None of the samples screened for microfilariae by Knott's test or for Dirofilaria immitis antigen by DiroChek^® were positive. Using PCR methods Leishmania-DNA was detected in 1/42 samples but none of 59 animals screened for E. canis-DNA was positive. The prevalence values as established by indirect and direct pathogen detection are considered as rather low.     

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.     

Combination of serum phosphorylated neurofilament heavy subunit and hyperintensity of intramedullary T2W on magnetic resonance imaging provides better prognostic value of canine thoracolumbar intervertebral disc herniation

The aim of this study was to evaluate the prognostic value of concurrent measurement of serum phosphorylated neurofilament heavy subunit (pNF-H) concentration and intramedullary T2W hyperintensity in paraplegic to paraplegic dogs. Our hypothesis was that concurrent measurement of these would provide a more accurate prediction of functional outcome in dogs with thoracolumbar intervertebral disc herniation (IVDH). A prospective case-control clinical study was designed using 94 dogs with acute onset of thoracolumbar IVDH. The association of serum pNF-H concentration, T2W hyperintensity on sagittal MRI (T2H/L2), deep pain perception and surgical outcome were evaluated with logistic regression analysis after three months for all 94 surgically treated dogs. Sensitivity to predict non-ambulatory outcome was compared among pNF-H and T2H/L2 and combination of both. Logistic regression analysis indicated that serum pNF-H concentration and T2H/L2 were significantly correlated with surgical outcome (P<0.05); however, deep pain perception was not (P=0.41). The results of logistic regression analysis indicated that the odds ratios of unsuccessful long-term outcome were 2.6 for serum pNF-H concentration, 1.9 for T2H/L2 and 2.3 for deep pain sensation. The sensitivity and specificity to predict non-ambulatory outcome for using serum parameter pNF-H>2.6 ng/ml, using T2H/L2 value of>0.84 and using both serum pNF-H and T2H/L2, were 95% and 75.7%, 65% and 86.5%, and 90.0% and 97.5%, respectively. Therefore, combined measurements of serum pNF-H and T2H/L2 might be useful for predicting long-term outcome in dogs with thoracolumbar IVDH.     

Analytical performance and method comparison of a quantitative point-of-care immunoassay for measurement of bile acids in cats and dogs

Point-of-care analyzers (POCAs) for quantitative assessment of bile acids (BAs) are scarce in veterinary medicine. We evaluated the Fuji Dri-Chem Immuno AU10V analyzer and v-BA test kit (Fujifilm) for detection of feline and canine total serum BA concentration. Results were compared with a 5th-generation assay as reference method and a 3rd-generation assay, both run on a bench-top analyzer. Analytical performance was assessed at 3 different concentration ranges, and with interferences. For method comparison, samples of 60 healthy and diseased cats and 64 dogs were included. Linearity was demonstrated for a BA concentration up to 130 µmol/L in cats (r = 0.99) and 110 µmol/L in dogs (r = 0.99). The analyzer showed high precision near the lower limit of quantification of 2 µmol/L reported by the manufacturer. Intra- and inter-assay coefficients of variation were < 5% for both species and all concentrations. Interferences were observed for bilirubin (800 mg/L) and lipid (4 g/L). There was excellent correlation with the reference method for feline (r_s = 0.98) and canine samples (r_s = 0.97), with proportional biases of 6.7% and −1.3%, respectively. However, a large bias (44.1%) was noted when the POCA was compared to the 3rd-generation assay. Total observed error was less than total allowable error at the 3 concentrations. The POCA reliably detected feline and canine BA in clinically relevant concentrations.