Dermatophagoides mites in house dust as an allergen source in atopic dogs

Thirty dogs with a clinical diagnosis of atopy were skin tested with 58 allergens including an aqueous house dust mite extract and a crude house dust extract. Sixty percent of the dogs had positive skin reactions to both the house dust mite and house dust antigens. In atopic dogs, house dust mites appear to be an important allergen source in house dust extracts but are not the only major source.     

Quantitation of house dust mites and house dust mite allergens in the microenvironment of dogs

OBJECTIVE: To quantitate the density of Dermatophagoides farinae and D pteronyssinus and concentrations of house dust mite (HDM) allergens (Der f 1, Der p 1, and Group 2 allergens) in the indoor microenvironment of dogs. SAMPLE POPULATION: 50 homes in Columbus, Ohio. PROCEDURES: In each home, samples of dust were collected from 3 locations in which dogs spent most time. Whenever possible, the species of mites collected was identified. Mite density (mites/g of dust) was assessed, and allergen concentrations were assayed by standardized ELISAs. Relative humidity and temperature in each home were monitored during a 5-day period. Characteristics of homes and sample sources were evaluated. RESULTS: Dust samples from all 50 homes contained > or = 1 HDM allergen; Der f 1 and Der p 1 were detected in 100 and 74% of homes, respectively. Fifteen homes had HDMs; compared with D pteronyssinus, D farinae was found more commonly (14/15 homes) and at a higher density. Basements, homes without central air-conditioning, and dog beds that were > or = 1 year old had high HDM allergen concentrations. Homes with > or = 2 microg of Der f 1 or Group 2 allergens/g of dust or > or = 100 mites/g of dust were significantly more likely to have a maximum relative humidity > or = 75%. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated the presence of HDMs and HDM allergens in the specific microenvironment of dogs in homes. Factors associated with high levels of exposure were identified, which may be associated with increased risk for sensitization and development of atopic diseases.     

Quantitation of house dust mites and house dust mite allergens in the microenvironment of dogs

OBJECTIVE: To quantitate the density of Dermatophagoides farinae and D pteronyssinus and concentrations of house dust mite (HDM) allergens (Der f 1, Der p 1, and Group 2 allergens) in the indoor microenvironment of dogs. Sample Population-50 homes in Columbus, Ohio. PROCEDURES: n each home, samples of dust were collected from 3 locations in which dogs spent most time. Whenever possible, the species of mites collected was identified. Mite density (mites/g of dust) was assessed, and allergen concentrations were assayed by standardized ELISAs. Relative humidity and temperature in each home were monitored during a 5-day period. Characteristics of homes and sample sources were evaluated. RESULTS: Dust samples from all 50 homes contained > or = 1 HDM allergen; Der f 1 and Der p 1 were detected in 100 and 74% of homes, respectively. Fifteen homes had HDMs; compared with D pteronyssinus, D farinae was found more commonly (14/15 homes) and at a higher density. Basements, homes without central air-conditioning, and dog beds that were > or = 1 year old had high HDM allergen concentrations. Homes with > or = 2 microg of Der f 1 or Group 2 allergens/g of dust or > or = 100 mites/g of dust were significantly more likely to have a maximum relative humidity > or = 75%. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate the presence of HDMs and HDM allergens in the specific microenvironment of dogs in homes. Factors associated with high levels of exposure were identified, which may be associated with increased risk for sensitization and development of atopic diseases.     

Sensitivity patterns to house dust mites and forage mites in atopic dogs: 150 cases

This study investigated intradermal test reactions to extracts of six species of mites in 150 dogs with atopic dermatitis. At least one positive reaction was seen in 120 animals (80%). Dermatophagoides farinae attracted the highest number of positive reactions (108 dogs, 90% of dogs and 72% of atopic dogs showing positive reactions). Positive reactions to other mites were not uncommon, with many dogs testing positive for Dermatophagoides pteronyssinus (32% of dogs tested), Acarus siro (35%), Tyrophagus putrescentiae (30%), Glycyphagus domesticus (27%) and Lepidoglyphus destructor (23%). Sensitivity to D. farinae alone occurred commonly (57% of cases), but multiple sensitivities were seen frequently with the other mites. Cases of sensitivity to only one mite were also seen: D. pteronyssinus (five cases), T. putrescentiae (one case) and G. domesticus (one case). Further studies are needed to appreciate more clearly the precise role played by the different species of mite in canine atopic dermatitis.     

The clinical effect of environmental control of house dust mites in 60 house dust mite-sensitive dogs

Abstract   The purpose of this study was to evaluate the effects of benzyl benzoate, an acaricide for the control of house dust mites, in 60 house dust mite-sensitive dogs. All dogs showed positive reactions on intradermal skin testing for house dust mites ( Dermatophagoides farinae , Dermatophagoides pteronyssinus ) alone, or house dust mites with storage mites ( Acarus siro , Tyrophagus putrescentiae , Glycophagus domesticus ). House dust samples from the owners' houses were collected and sent to the clinic, where the authors performed a test (Acarex® test) to semiquantify the amount of guanine, a house dust mite product. Treatment with benzyl benzoate was repeated until the house dust samples were negative for house dust mite guanine. After treatment, 29 out of 60 house dust mite-sensitive dogs (48%) showed no skin lesions or pruritus. Moderate results were achieved in 22 dogs (36%), with reduced pruritus and minimal skin lesions, but still requiring medication. In 13 dogs, this involved regular treatment (3–4 times a year) with antibiotics and antiyeast medication, and in eight dogs, immunotherapy was used. One dog was controlled with essential fatty acids as monotherapy and one dog was controlled with immunotherapy and essential fatty acids. In the remaining nine dogs (15%), the pruritus remained the same, and these dogs were controlled with oral corticosteroids. These results indicate that house dust mite elimination is a useful tool in the management of house dust mite-sensitive dogs.     

Patch test reactions to house dust mites in dogs with atopic dermatitis

The purpose of this study was to determine the percentage of dogs with spontaneous atopic dermatitis that show a positive patch test reaction to a commercially available 20% house dust mites mixture containing equal parts of Dermatophagoides farinae and Dermatophagoides pteronyssinus in white petrolatum. In addition, we evaluated whether skin reactions induced after the epicutaneous application of house dust mites were clinically and histologically similar to naturally developed skin lesions of dogs with atopic dermatitis. Furthermore, we investigated if the reactions induced by house dust mites were true allergic reactions by comparing them to atopic lesional skin and to patch test reactions induced by an irritant substance (sodium lauryl sulphate). White petrolatum alone and nonlesional skin sites were used as negative controls. Macroscopic and microscopic evaluations of the patch test and control sites were performed in a blinded fashion at 48 and 72 h after patch test application. Microscopic results were evaluated in a qualitative and quantitative manner. A chi‐square test for homogenicity was used for the quantitative analysis to compare the proportion of each dermal inflammatory cell type among positive histopathological tested sites. P values ≤ 0.05 were considered significant. The study included 12 healthy nonatopic dogs and 13 dogs with nonseasonal atopic dermatitis. None of the nonatopic dogs reacted to house dust mites and white petrolatum. Ten (77%) of the 13 atopic dogs reacted macroscopically and histopathologically to house dust mites. Macroscopic reactions induced by house dust mites were characterized by erythema, oedema and papules. The macroscopic reactions induced by house dust mites were identical to lesional skin in 20% of the dogs and identical to reactions induced by sodium lauryl sulphate in 40% of the dogs. Qualitative histopathological findings showed that the reactions induced by house dust mites were similar to atopic lesional skin in 80% of the dogs and were similar to sodium lauryl sulphate in 20% of the dogs. Quantitative analyses showed that the proportion of neutrophils in reactions induced by sodium lauryl sulphate was significantly higher (P < 0.05) compared to house dust mites reactions, which could be a differentiator factor between an allergic and an irritant reaction. These results showed that the epicutaneous application of house dust mites in dogs with atopic dermatitis induced histopathological lesions similar to spontaneous atopic lesions in dogs. Therefore, this study demonstrated that house dust mites penetrated the skin of dogs with atopic dermatitis and induced an inflammatory response that resembled a true allergic reaction. Funding: Small Companion Animal Grant, University of Minnesota.     

Effects of age and allergen exposure on transepidermal water loss in a house dust mite-sensitized beagle model of atopic dermatitis

Transepidermal water loss (TEWL) was measured as assessment of barrier function in house dust mite-sensitized atopic ( n  = 24) and normal ( n  = 21) beagle dogs before and after allergen challenge. Dogs of the two groups were matched for age and housed in the same environment. Ten sites were evaluated including both areas of high and low predisposition to atopic lesion formation. On day 0 (before allergen exposure) and day 3 (3 days after allergen exposure), TEWL measurements were taken using an open-chamber evaporimeter. On day 0, significant differences (one-way ANOVA) were found between normal and atopic dogs in the chin, pinna, periocular region, axilla, antebrachium and thorax. Within the atopic group, a significant increase (one-way ANOVA) in TEWL was found in the axilla on day 3. Such a difference was particularly marked in young dogs (chin, pinna, axilla, antebrachium and groin). Within the normal group, allergen challenge did not change TEWL except for the chin. When atopic and nonatopic sites were compared within the atopic group, a significant difference in TEWL was found on day 0. A two-way ANOVA showed a significant effect of site, a significant effect of time but no site × time (i.e. allergen exposure) interaction. It is concluded that the house dust mite-sensitized atopic beagle dogs used in this study have altered barrier function in 'atopic' areas and that such differences are more evident in young individuals and are aggravated by allergen exposure.     

FC‐30 Presence of dust mites in the environment of dust mite‐sensitized atopic dogs

Dust mites (DM) are the most common offending aeroallergens in atopic dogs. The aim of this study was to compare the DM load of households with atopic dogs (Group A, n = 8) that had positive intradermal test reactions to Dermatophagoides farinae, D. pteronyssinus, Acarus siro, Lepidoglyphus destructor and/or Tyrophagus putrescentiae to the DM load of households with nonatopic dogs (Group B, n = 4) and of nonpet households (Group C, n = 8). Group A dogs presented with perennial pruritus, were free of pathogenic mites and fleas, did not respond to an elimination diet, and fulfilled the diagnostic criteria of atopic dermatitis. All Group B dogs tested intradermally negative and had no dermatological problems. Dust samples were vacuum collected in a standardized fashion from the human (all groups) and dog mattresses (Groups A and B) or from the couch (Group C) four times, once for each season of the year. The presence of DM was assessed with a commercial test (Acarex test) and stereoscopically. At least one DM was found in all Group A houses. The DM load was not significantly different between the seasons or the three animal groups. The sensitivity of the Acarex test was significantly lower than that of stereoscopic examination (P < 0.001). In conclusion, the environmental DM load was similar between atopic and nonatopic dogs, the presence of dogs in a household didn't increase DM numbers, and stereoscopy was more sensitive than the Acarex test for the detection of DM. Funding: Self‐funded.     

Determination of skin threshold concentration of an aqueous house dust mite allergen in normal dogs

Fifty-six healthy dogs with no known history of atopic disease to indoor allergens were skin tested with 6 different dilutions of an aqueous house dust mite extract. A concentration of 31.25 PNU/ml was found to be the maximum, nonirritating concentration. Thirty-two percent of the research dogs used had to be excluded because they failed to show reactions to any test dilution.     

Evaluation of the presence of house dust mites in horse rugs

A sample of fourteen horse rugs and two saddle blankets stored in south western Sydney, Australia, an area of known high dust mite prevalence in the human environment, were analysed for the presence of house dust mites. Dust samples from the rugs, blankets and 16 control sites were collected using a vacuum cleaner with a modified attachment and filter. Dust mites were extracted using an adapted floatation technique. Eight rugs and all control samples were positive for mites, which were confirmed to be house dust mites of the genus Dermatophagoides. This study confirms that exposure to house dust mites from horse rugs can occur, indicating that house dust mite allergen reactivity on intradermal and serum allergy testing in atopic horses may represent true dust mite hypersensitivity. Nevertheless, quantification studies will be necessary to ensure that there is adequate mite exposure for development of hypersensitivity, and further evaluation of immunological responses, avoidance and provocation, and specific immunotherapy are required to confirm the clinical relevance. To the authors’ knowledge, this is the first study to document the presence of house dust mites in the equine environment.     

Characterization and cloning of a major high molecular weight house dust mite allergen (Der f 15) for dogs

Although house dust mites (HDM(s)) are important elicitors of canine allergy, the low molecular weight molecules defined as major allergens for humans do not appear to be major allergens for dogs. Western blotting of Dermatophagoides farinae (D. farinae) extracts with sera from sensitized dogs showed that the majority of animals had IgE antibodies specific for two proteins of apparent molecular weights of 98 and 109kDa (98/109kDa). The N-terminal sequences of these two proteins were identical, suggesting they were very closely related, and sequencing of internal peptides showed the protein(s) to have homology with insect chitinases. A purified preparation of 98/109kDa proteins elicited positive intradermal skin tests (IDST(s)) in a group of well-characterized atopic dogs sensitized to D. farinae, but not in normal dogs. A rabbit polyclonal antiserum raised against the purified proteins was used to immunoscreen a D. farinae cDNA library. The mature coding region of the isolated chitinase cDNA predicts a protein of 63.2kDa; sequence analysis and glycan detection blotting suggest that the molecule is extensively O-glycosylated. Monoclonal antibodies made against the purified native protein were used to localize the chitinase in sections of whole D. farinae mites. The protein displayed an intracellular distribution in the proventriculus and intestine of the mite, suggesting that it has a digestive, rather than a moulting-related, function. The high prevalence of IgE antibodies to this antigen in canine atopic dermatitis makes it a major HDM allergen for dogs, and the protein has been formally designated Der f 15.     

FC-31 Prevalence and characterization of house dust mites and house dust mite allergens collected from the bedding, skin and hair coat of dogs in southwest England

Dust mites (DM) are the most common offending aeroallergens in atopic dogs. The aim of this study was to compare the DM load of households with atopic dogs (Group A, n  = 8) that had positive intradermal test reactions to Dermatophagoides farinae , D. pteronyssinus, Acarus siro, Lepidoglyphus destructor and/or Tyrophagus putrescentiae to the DM load of households with nonatopic dogs (Group B, n  = 4) and of nonpet households (Group C,  n  = 8). Group A dogs presented with perennial pruritus, were free of pathogenic mites and fleas, did not respond to an elimination diet, and fulfilled the diagnostic criteria of atopic dermatitis. All Group B dogs tested intradermally negative and had no dermatological problems. Dust samples were vacuum collected in a standardized fashion from the human (all groups) and dog mattresses (Groups A and B) or from the couch (Group C) four times, once for each season of the year. The presence of DM was assessed with a commercial test (Acarex test) and stereoscopically. At least one DM was found in all Group A houses. The DM load was not significantly different between the seasons or the three animal groups. The sensitivity of the Acarex test was significantly lower than that of stereoscopic examination ( P  < 0.001). In conclusion, the environmental DM load was similar between atopic and nonatopic dogs, the presence of dogs in a household didn't increase DM numbers, and stereoscopy was more sensitive than the Acarex test for the detection of DM.
Funding: Self-funded.     

Cellular and cytokine kinetics after epicutaneous allergen challenge (atopy patch testing) with house dust mites in high-IgE beagles

The cellular and cytokine dynamics of reactions triggered by atopy patch testing with house dust mites were studied in six high-IgE beagles. Sites were scored and biopsied at 6, 24, 48, and 96 h, and samples were processed for histopathology, immunohistochemistry, and polymerase chain reaction (PCR). All dogs developed positive reactions at some point in time. Mean clinical scores were significantly higher than baseline at 24, 48, and 96 h. Clinically, one of six dogs had a positive reaction at 6 h; two of six reacted at 24 and 48 h, and five of six at 96 h. Histologically, superficial perivascular mononuclear and granulocytic dermatitis developed (5/6) after 6 h, and progressed in severity at 24 h (6/6). Additionally, at 48 h epidermal spongiosis, hyperplasia and pustules developed (5/6), and were marked at 96 h (6/6). At and beyond 6 h, progressive CD1c-positive epidermal Langerhans cell hyperplasia with cluster formation and dermal dendritic cell infiltration was noted. Cutaneous infiltration of CD3-positive T lymphocytes with epidermal clusters developed over time. mRNA expression for the cytokines gamma-interferon (gamma-IFN), interleukin-6 (IL-6), IL-12p35, IL-13, IL-18, and thymus and activation regulated chemokine (TARC) exhibited significant increases during the challenge compared to baseline, but there was no appreciable alteration in expression for tumour necrosis factor-alpha (TNF-alpha), IL-12p40, IL-10, regulated on activation normal T-cell expressed and secreted (RANTES), IL-5, IL-2, IL-4, and IL-8. No correlation was detected between clinical scores and cytokines. It is concluded that IL-6 plays a role in early reactions followed by an increase of TARC and IL-13, while IL-18 progressively increases in later reactions.     

Clinicopathologic findings, sensitivity to house dust mites and efficacy of milbemycin oxime treatment of dogs with Cheyletiella sp. infestation

Twenty-three dogs with positive skin scrapings for Cheyletiella sp. were treated with milbemycin oxime using a protocol approximating 2 mg kg⁻¹ orally once weekly for three weeks. Nineteen of these dogs belonged to a household of 41 dogs and two dogs were in households with one other dog. All in-contact dogs were treated. Pre-treatment intradermal skin tests showed positive reactions to D. farinae in 13 dogs and to D. pteronyssinus in 12 dogs; these became negative post-treatment in four and seven dogs, respectively. All dogs showed a dramatic reduction in clinical signs one week after the third treatment. Eighteen dogs no longer had mites on skin scrapings, three had dead mites and two had deformed eggs. Recurrence of clinical signs necessitated two additional courses of the protocol in the multiple dog household and for a dog receiving immunosuppressive treatment for pemphigus foliaceus. Possible adverse reactions to the milbemycin (vomiting, lethargy) were noted once in two dogs.     

Seasonal dynamics of house dust mites in dust samples collected from sleeping places in north-western Poland

The most common families of mites found in house dust are Pyroglyphidae, Glycyphagidae and Acaridae; all are a source of many antigens responsible for allergic diseases. The aim of this study was to examine the seasonal dynamics of allergenic mite populations in dust samples collected from sleeping places in apartments in north-western Poland. The mites were isolated from the dust using a saturated saline floating method. In 132 dust samples we determined: Dermatophagoides farinae, Dermatophagoides pteronyssinus, Euroglyphus maynei, Hirstia sp., Chortoglyphus arcuatus, Lepidoglyphus destructor, Gohieria fusca and Cheyletus sp. The greatest frequency was observed for D. farinae, D. pteronyssinus, Ch. arcuatus and Cheyletus sp., in the fourth quarter and D. farinae in the third quarter. Smaller coefficients of dominance were found for D. pteronyssinus, Ch. arcuatus and Cheyletus sp., and their greatest mean concentrations were found in the first and fourth quarters. Given the division of the year into heating and non-heating seasons, mites D. farinae and D. pteronyssinus achieved the highest mean concentration in the first season, and Cheyletus sp. in the second season. The analysis of the participation of developmental stages showed that the adults of D. farinae were more prevalent than juveniles in the first, second and third quarters, and imago stages of D. pteronyssinus were more numerous in relation to juveniles in the first, third and fourth quarters. The results confirm the high incidence of house dust mites in sleeping places in north-western Poland dwellings; the best conditions for the development of these mites, mainly D. farinae and D. pteronyssinus, occur in the fourth quarter and are the least favourable in the second quarter. In many cases, these results are consistent with data from other parts of Poland collected by various authors.     

Atopy patch test reactions in high-IgE beagles to different sources and concentrations of house dust mites

Protocols for atopy patch testing (APT) were evaluated on six high-IgE dogs sensitized to house dust mites (HDM) using various concentrations and sources of HDM. Two sources of HDM were compared: Heska slurry and four concentrations of Greer HDM. Saline was used as a negative control. Patches were removed after 48 h and the sites evaluated at 0, 6, 24, 48, 72 and 96 h for erythema, macules, papules and pustules. Each sign was scored from 0 to 3 (0 = absent and 3 = severe). Total score was used for analysis. Mean total scores significantly increased for both Greer and Heska HDMs from 6 h, peaking at 48 h for G100 (100 mg mL(-1)), G300 and G668, and at 72 h for Heska and G31.25. Across all times, Heska HDM scores were significantly higher than those of G31.25 with the largest difference at 96 h. Heska scores, however, were significantly lower than those of other Greer concentrations (G100, G300 and G668) particularly at 96 h. No reactions were noted at saline sites. It is concluded that Greer-HDM at 100 mg mL(-1) is the most suitable concentration for APT in dogs because it induces reactions comparable if not higher than more concentrated HDM preparations.     

Effects of calcium supplementation and sunlight exposure on growing beagle dogs

Nine 7-week-old Beagle dogs were given 2.3 g of dietary calcium supplementation once a day and were exposed to sunlight to permit endogenous vitamin D formation. After 70 days, the serum calcium and phosphorus concentrations, thyroid morphometry, and bone density and composition were compared with those values in a similar group of 10 dogs not given calcium supplementation and maintained in restricted, artificial light. Lower serum calcium and phosphorus concentrations in the supplement-fed dogs may be attributed to hypercalcitonism, which was assumed to be present because of increased proportions of C cells and decreased proportions of thyroid follicles. They also had significantly more trabecular bone of a lower specific gravity containing more water and less combustible matter. The findings indicate that the high dietary calcium intake and sunlight exposure induced C-cell hyperplasia during a relatively short period of 70 days. It is possible that Beagles maintained through maturity on a diet high in calcium concentration and in outdoor runs may develop chronic C-cell hyperplasia. The possible contribution of high calcium intake and chronic C-cell hyperplasia to reduced thyroid function and skeletal changes is not yet clear.     

Use of an amplified ELISA technique for detection of a house dust mite allergen (Der f 1) in skin and coat dust samples from dogs

OBJECTIVE: To use an amplified ELISA technique to document the presence and quantify the concentration of the house dust mite allergen, Der f 1, in skin and coat dust samples collected from dogs. ANIMALS: 29 pet dogs of various breeds. PROCEDURE: Dogs were weighed, and body surface area in square meters was determined. Skin and coat dust samples were obtained by vacuuming dogs. Collected dust was analyzed by use of standard and amplified ELISA techniques. RESULTS: By use of the standard ELISA technique, Der f 1 was detected in skin and coat dust samples from 6 of 29 (21%) dogs. Mean concentration of Der f 1 in the 6 samples with positive assay results was 16.16 ng/mL (range, 5.61 to 31.24 ng/mL). Samples with negative assay results were retested for dust mite allergen by use of an amplified ELISA technique; an additional 14 dogs had positive assay results. Mean concentration of allergen was 0.36 ng/mL (range, 0.19 to 2.20 ng/mL). Combining both techniques, 20 of 29 (69%) dogs had positive assay results for Der f 1. CONCLUSIONS AND CLINICAL RELEVANCE: Results of our study indicate that house dust mite allergens are present on the skin and in the coat of dogs, and this source of allergen may act as a reservoir for allergen exposure in hypersensitive dogs. Use of an amplified ELISA technique to determine environmental concentrations of house dust mite allergens in homes and on dogs will help to identify the relationship between immunologic findings and environmental exposures in dogs with atopic dermatitis.