Investigation of bacterial microorganisms in the conjunctival sac of clinically normal dogs and dogs with ulcerative keratitis in Beijing, China

Objective To determine the bacterial microorganisms in the conjunctival sac of clinically normal dogs and dogs with ulcerative keratitis in Beijing, China. The effect of breed, sex and age of dogs and season on the presence or absence of bacteria in the conjunctival sac of clinically normal dogs was evaluated. Sample population This investigation included 240 healthy dogs, 27 dogs with unilateral corneal ulcer and one dog with bilateral corneal ulcer. Procedure The 480 samples from healthy dogs and the 29 samples from dogs with ulcerative keratitis were incubated in an aerobic and 5% CO₂ environment at 37 °C for 48 h. Logistic regression analysis was performed. Statistical significance was set at P < 0.01. Results Of 480 normal eyes, Staphylococcus spp. were the most frequently isolated organisms (40.29%). Neisseria spp. (11.47%) were the next most frequently isolated organisms, followed by Corynebacterium spp. (9.4%). Of 29 eyes with ulcerative keratitis, Staphylococcus spp. were also the most frequently isolated bacteria (47.06%). Streptococcus spp. (12.94%) and Pseudomonas spp. (8.24%) were the second and third, respectively. Season (P = 0.002) was a significant factor influencing presence or absence of bacterial microorganisms in the conjunctival sac of normal dogs in Beijing, China, while the effects of breed (P = 0.095), sex (P = 0.588) and age (P = 0.866) of dogs were insignificant. Conclusion Staphylococcus spp. were the most frequently isolated organisms, and S. intermedius predominated in the conjunctival sac of clinically normal dogs and dogs with ulcerative keratitis in Beijing, China. The likelihood of detecting bacteria depends on the season.     

Erythrocyte and plasma fatty acid patterns in dogs with atopic dermatitis and healthy dogs in the same household

Recent studies have indicated that dogs with canine atopic dermatitis (CAD) may have a disorder of fatty acid metabolism: possibly low or absent activity of delta6-desaturase or delta5-desaturase, or both. To clarify this possibility, we examined the erythrocyte and plasma fatty acid patterns of 8 dogs with CAD and their 8 healthy housemates. Atopic dermatitis was diagnosed according to the criteria proposed by Willemse; other causes of dermatitis were excluded clinically and by appropriate tests. Erythrocyte ghosts were prepared from blood samples. Membrane lipids were extracted and separated by thin-layer chromatography. From plasma and lipid fractions, fatty acid content was determined by gas chromatography. In erythrocytes, but not in plasma, we observed significant differences in the fatty acid pattern that suggested a reduction in the n6 fatty acid products of the delta6- and delta5-desaturases in dogs with atopic dermatitis when compared with healthy housemates.     

Serial thyroid hormone concentrations in healthy euthyroid dogs, dogs with hypothyroidism, and euthyroid dogs with atopic dermatitis.

Serum thyroxine (T4) and 3,5,3'-triiodothyronine (T3) concentrations were determined every 3 h for 12 h beginning at 8 a.m. in 20 healthy euthyroid dogs, 19 dogs with hypothyroidism, and 18 euthyroid dogs with atopic dermatitis. Status of thyroid function was based on history, physical findings, results of thyrotropin response testing, and requirement for thyroid hormone replacement therapy. Mean serum T4 and T3 concentrations did not vary significantly between blood samplings within each of the three groups of dogs. Between groups of dogs, mean serum T4 concentration was significantly (P less than 0.05) higher at each blood sampling time in healthy euthyroid dogs and euthyroid dogs with atopic dermatitis when compared to dogs with hypothyroidism. There was no significant difference in mean serum T4 concentration at any blood sampling time between healthy euthyroid dogs and euthyroid dogs with atopic dermatitis or in mean serum T3 concentrations at any blood sampling time between any of the three groups of dogs. Random fluctuation in serum T4 and T3 concentrations was found in dogs in all three groups. Random fluctuations were more common with serum T3 versus T4 concentrations. Consequently, sensitivity (0.88 versus 0.52), specificity (0.73 versus 0.45), predictive value for a positive test (0.75 versus 0.32), predictive value for a negative test (0.87 versus 0.65), and accuracy (0.80 versus 0.47) were better for serum T4 concentration than serum T3 concentration, respectively, when all blood samples were analysed. Measurement of serum T4 concentration was more accurate than serum T3 concentration in assessing the status of thyroid gland function.     

Late-phase reactions to intradermal testing with Dermatophagoides farinae in healthy dogs and dogs with house dust mite-induced atopic dermatitis

OBJECTIVE: To determine the prevalence of late-phase reactions to intradermal testing with Dermatophagoides farinae in healthy dogs and dogs with atopic dermatitis and an immediate reaction to D farinae. ANIMALS: 6 healthy dogs and 20 dogs with atopic dermatitis and immediate reactions to D farinae. PROCEDURE: ntradermal tests were performed with D farinae at 1:1,000 wt/vol and 1:50,000 wt/vol concentrations, and skin reactivity was evaluated after 0.25, 6, and 24 hours. Serum D farinae-specific IgE antibodies were assayed. Extent of lesions (atopy index) and pruritus (visual analogue scale) were evaluated in dogs with atopic dermatitis. RESULTS: Late-phase reactions were observed in healthy dogs at 6 hours (n = 2 dogs) and 24 hours (1) with the 1:1,000 wt/vol concentration, and at 6 hours (1) and 24 hours (1) with the 1:50,000 wt/vol concentration of allergen. Late-phase reactions in healthy dogs were only observed in dogs with an immediate reaction to D farinae. Late-phase reactions were observed in 11 of 20 dogs with atopic dermatitis at 6 and 24 hours with the 1:1,000 wt/vol concentration and in 10 of 20 at 6 and 24 hours with the 1:50,000 wt/vol concentration of allergen. There was no difference in mean atopy index, mean visual analogue scale of pruritus, or mean serum D farinae-specific IgE concentration of dogs with a late-phase reaction, compared to dogs without a late-phase reaction. CONCLUSIONS AND CLINICAL RELEVANCE: Late-phase reactions may be observed after an immediate reaction to intradermal skin testing in healthy and allergic dogs but are more commonly observed in dogs with atopic dermatitis.     

Bacterial and mycoplasmal flora of the healthy camelid conjunctival sac

Healthy conjunctival sacs of 88 animals of 3 species of captive camelids (Lama glama, Lama guanicoe, Lama pacos) and llama-guanaco hybrids were sampled for bacterial and mycoplasmal flora. Mycoplasmas were not isolated from any animal. Eleven genera of bacteria were isolated. The most frequent isolates were Staphylococcus epidermidis and Pseudomonas spp. Nine varieties of Pseudomonas were found, which represented at least 3 Pseudomonas species. Many of the bacterial isolates (especially the pseudomonads) are potential pathogens in the eyes of these camelids.     

Fungal flora of the healthy camelid conjunctival sac.

Swab specimens for fungal isolation were collected from the healthy conjunctival sacs of 3 species of captive camelids (Lama glama, L guanicoe, L pacos) and llama-guanaco hybrids. Fungi were collected from over half the animals in winter (53%) and summer (56%). Fungal species of 10 genera were isolated. In both seasons, Aspergillus was the most commonly isolated genus; at least 9 species of Aspergillus were found. The fungal organisms isolated were similar to those found in healthy eyes of other domestic animals and may represent a random seeding from the environment where they are ubiquitous.     

Allergic conjunctivitis and conjunctival provocation tests in atopic dogs

Introduction Canine atopic dermatitis (cAD) is a very common disease, but little is known about eye involvement. The conjunctival provocation test (CPT) is used in human to study the ocular response to allergenic stimuli and to evaluate anti‐allergic therapy. To our knowledge it has not been used in dogs. Objectives To evaluate the prevalence of ocular signs in a population of atopic dogs and relate these with clinical cAD scores; and the usefulness of CPT for dust mites in atopic dogs with itchy eyes. Procedures Sixty cAD patients were evaluated for (i) ocular signs of allergic conjunctivitis including conjunctival hyperemia, chemosis, epiphora, ocular discharge, pruritus and corneal involvement, graded 0 to 3 according to severity, and (2) cAD Extent and Severity Index (CADESI‐03). Additionally, CPTs for Dermatophagoides farinae (n = 12) and Dermatophagoides pteronyssinus (n = 12) were performed in sensitized atopic dogs and 24 control dogs. Results Periocular and ocular signs of allergy were present in 60% (36/60) of cases. Conjunctival hyperemia (90%) was the most common sign. Severity of ocular signs was significantly correlated with eye pruritus (r_s = 0.690, P = <0.001) and skin lesions score for head region (r_s = 0.261, P = 0.04). A highly significant difference (P < 0.001, Fisher test) was found in CPTs between the test and the control groups. Conclusion Allergic conjunctivitis signs associated with cAD seem under valuated so these patients would benefit from an ophthalmologic evaluation. Furthermore, we found CPT to be a reliable, easy to perform and safe test for the etiologic diagnosis of allergic conjunctivitis in the dog.     

Evaluation of skin test reactivity to environmental allergens in healthy cats and cats with atopic dermatitis

OBJECTIVE: To evaluate skin test reactivity to environmental allergens in healthy cats and in cats with atopic dermatitis (AD). ANIMAL: 10 healthy cats and 10 cats with AD. PROCEDURE: 10 allergens in serial dilutions were injected ID on the lateral aspect of the thorax of sedated cats. Histamine (0.01% solution) and buffer solutions were used as positive and negative controls, respectively. Immediately after the last injection, 10% fluorescein solution was administered IV. Skin test results were evaluated with ultraviolet light after 15 to 30 minutes and at 4 and 6 hours by 2 independent observers. In the control group, skin tests were repeated after 6 weeks. Skin test reactivity and the nature of the immunoglobulin involved were investigated by use of the Prausnitz-Küstner test with untreated and heat-treated cat sera. RESULTS: Intertest and interobserver agreement were high when measurement of the diameter of the fluorescent wheal was used to evaluate skin test responses, compared with assessment of its intensity. In both groups of cats, immediate skin test reactivity was observed as an IgE-mediated reaction, as an IgG-mediated reaction, and as a result of nonspecific mast cell degranulation. There was no correlation between allergen concentration and the type of reaction observed. CONCLUSIONS AND CLINICAL RELEVANCE: Skin test reactivity in cats should be evaluated after IV administration of 10% fluorescein solution by means of a Prausnitz-Küstner test to differentiate among IgE-mediated, IgG-mediated, and nonspecific reactions.     

Results of allergen-specific immunotherapy in 117 dogs with atopic dermatitis

The success of the treatment of 117 dogs with atopic dermatitis with allergen-specific immunotherapy for up to 48 months was assessed. An excellent response (remission with exclusive immunotherapy) was recorded in 18 of the dogs, a good response (more than 50 per cent reduction in medication and improvement of clinical signs) was recorded in 57, a moderate response was recorded in 24 and a poor response in 18. The mould antigens in the allergen extract were stored in a separate vial before administration and the success rate of the immunotherapy including mould antigens was much higher than in an earlier study in which mould and pollen antigens had been stored in one vial. The success rate was not affected significantly by the age of the dogs when the disease developed, or by their age or the period for which they had shown clinical signs when the treatment began; it was also unaffected by whether pollens, moulds or dust mites were used as antigens, or by whether the offending allergens had been identified by intradermal testing or by serum testing for allergen-specific immunoglobulin E.     

Dust mite species in the households of mite-sensitive dogs with atopic dermatitis

Background – The presence of important house dust and storage mite species in the microenvironment of atopic dogs has not been thoroughly investigated. Objectives – To compare the presence and population of five dust mite species (Dermatophagoides farinae, Dermatophagoides pteronyssinus, Acarus siro, Tyrophagus putrescentiae and Lepidoglyphus destructor) among households with mite‐sensitive atopic dogs (Group A), households with clinically healthy dogs (Group B) and households without pets (Group C, n = 25) in Greece. Animals – Twenty mite‐sensitive atopic dogs and 20 clinically healthy dogs. Methods – Dust samples were collected with a vacuum cleaner from owners’ mattresses (all groups) and from dogs’ sleeping areas (Groups A and B) or living room couch (Group C), once every season of the year. Following dust flotation, mites were counted and identified. Results – Dermatophagoides farinae was the most prevalent (60, 40 and 64% in Groups A, B and C, respectively), followed by D. pteronyssinus (45, 35 and 48%, respectively), whereas the three storage mites were found in fewer households. No major differences could be found between Groups A and B or between households with (Groups A and B) and without dogs (Group C) regarding the presence or numbers of the five dust mite species. Conclusions and clinical importance – The presence and population of five common house dust and storage mite species does not differ among Greek households with mite‐sensitive atopic dogs, households with healthy dogs and households without pets.     

Effect of hyposensitization on atopic dermatitis in dogs

In a double-blind study, 51 dogs with clinically defined atopic dermatitis were injected with either alum-precipitated allergen solutions or a placebo. Comparing the treatment results of both groups on the basis of scores for clinical signs, a significant difference in clinical improvement was established in favor of the allergen-treated dogs (P less than 0.01). The proportional changes of scores for clinical signs in the allergen-treated group ranged between +27.3% and -100% (median, -61.5%) and in the placebo group between +36.4% and -100% (median, 0.0%) with respect to the initial scores. Immediate skin test reactivity disappeared only in the dogs with a good clinical response. Of 27 dogs treated with an allergen solution, 16 (59.3%) had an improvement of 51% or more. In the placebo group, 5 of 24 dogs (20.8%) reacted this way. There was total remission of the clinical signs in 9 and 4 dogs, respectively. In the dogs in which, after 9 months of hyposensitization, any improvement was observed, the chance for final improvement of more than 51% was calculated as 84%. Discriminant analysis revealed that evaluation of the effect of immunotherapy can be restricted to the 9-month follow-up examination.     

Intradermal testing with the storage mite Tyrophagus putrescentiae in normal dogs and dogs with atopic dermatitis in Colorado

The purpose of this study was to evaluate reactions to intradermal injections of Tyrophagus putrescentiae extract in healthy dogs and dogs with atopic dermatitis and to compare the prevalence of positive reactions in the two groups. Twenty-one healthy dogs and 26 atopic dogs were tested intradermally with T. putrescentiae extract at 1000, 500, 250, 125, 63, 32 and 16 PNU/mL. Reactions were evaluated objectively and subjectively. A Mann-Whitney test was used to determine differences in grade of reaction to storage mites between healthy dogs and dogs with atopic dermatitis. Positive reactions to storage mite extract were most common at 1000 PNU/mL with approximately one third of normal and atopic dogs showing a positive reaction to T. putrescentiae. There was no significant difference in the incidence of positive reactions between normal and atopic dogs for any of the Tyrophagus extract concentrations.     

Evaluation of IgG subclass responses against Dermatophagoides farinae allergens in healthy and atopic dogs

A semiquantitative chemiluminescent Western blot analysis system was developed and validated to evaluate antigen-specific IgG subclass responses to electrophoretically separated proteins of Dermatophagoides farinae in healthy and atopic dogs. Both groups mounted similar D. farinae-specific IgG1 and IgG4 responses to multiple antigens, but IgG2 and IgG3 responses were difficult to detect. The most commonly recognized bands in both groups were 18 and 98 kDa antigens for IgG1 and 18, 45, 66, 98, 130 and 180 kDa for IgG4. The number of bands recognized per dog did not differ significantly, but significantly more atopic dogs had an IgG1 response to a 180 kDa protein. The overall D. farinae-specific IgG1 and IgG4 responses were slightly higher, but not significantly different, in the healthy group. The results suggest that some antigens produced by D. farinae can induce different subclass responses. However, as most of these responses are seen in both healthy and atopic dogs, they are likely to merely represent recognition of foreign proteins presented to the immune system, rather than involvement in the pathogenesis of atopic dermatitis. The role of the 180 kDa antigen warrants further study.     

Clinical trial evaluating the efficacy and safety of cyclosporine in dogs with atopic dermatitis

OBJECTIVE: To determine efficacy and safety of cyclosporine in the treatment of atopic dermatitis among dogs in North America. DESIGN: Randomized controlled (phase 1) and open-label (phase 2) trials. ANIMALS: 268 dogs with atopic dermatitis. PROCEDURE: In phase 1, dogs were randomly assigned to be treated with cyclosporine (5 mg/kg [2.3 mg/Ib], PO, q 24 h) or a placebo. In phase 2, all dogs were treated with cyclosporine for 16 weeks. Frequency of cyclosporine administration was decreased if dogs improved clinically. RESULTS: At the end of phase 1, canine atopic dermatitis extent and severity index (CADESI) scores for dogs treated with cyclosporine were significantly lower than scores for control dogs. Percentage of dogs with severe pruritus decreased from 67% to 16% for the cyclosporine group but from 66% to only 61% for the control group. During phase 2, cyclosporine dosage was decreased to every-other-day administration in 39% of the dogs after 4 weeks. After 12 weeks, 22% of the dogs were treated twice weekly and 36% were treated every other day. After 16 weeks, CADESI score had decreased > 50% in 68% of the dogs and 47% of dogs had no or mild pruritus. The most frequent adverse reactions were gastrointestinal tract signs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cyclosporine is efficacious for the treatment of atopic dermatitis in dogs and that frequency of cyclosporine administration can be reduced following an initial induction period. The drug was well tolerated.