Cutaneous epitheliotropic T-cell lymphoma in a donkey - a case report.

BACKGROUND: Cutaneous epitheliotropic T-cell lymphoma is a malignant tumour of the skin already reported in humans, dogs, cats, horses, and other species, but not previously in donkeys. The standard diagnosis is based on clinical, morphological and immunophenotypic data. Differentiation of malignant versus benign proliferation of lymphocytes is crucial; in ambiguous cases T-cell receptor gamma (TRG) molecular clonality should be tested. In the present paper, we report a case of mycosis fungoides diagnosed in a donkey whose diagnosis was based on clinical, histological and immunohistochemical aspects and a positive TRG clonality test. CASE PRESENTATION: A twenty-five-year-old donkey gelding was referred with a mildly pruritic, generalised and severe exfoliative dermatosis. Otherwise, the animal was clinically healthy, though mildly underweight. Dermatological examination revealed severe generalised alopecic and exfoliative dermatitis, occasionally eroded, with high number of large, thin, greyish scales. All mucocutaneous junctions except the hoofs were affected. Ectoparasites and dermatophytes were ruled out. The complete blood count and blood smear evaluation revealed mild normocytic normochromic anemia. The biochemistry panel showed mild hyperproteinemia with albumin within the normal range. Protein electrophoresis showed moderate polyclonal hypergammaglobulinemia. Histological findings were characterised by interface dermatitis with massive exocytosis in the epidermis of a homogenous population of lymphoid cells showing atypia. Clusters of neoplastic cells were present within the epidermis forming Pautrier "microabscesses". These findings are consistent with cutaneous epitheliotropic lymphoma. Immunohistochemical staining revealed uniform labelling of the neoplastic cells for CD3, and lack of expression of CD20 (a B cell lineage associated marker). Molecular clonality PCR (PARR) was performed using equine TRG primers; this revealed a clonal rearrangement in a heavy polyclonal background. Transmission electronic microscopy showed multiple lymphocytes with convoluted or cerebriform nuclei. CONCLUSIONS: This case report provides the first evidence of clinical, histopathological, immunophenotypic features, electron microscopy findings and molecular analysis of a cutaneous epitheliotropic T-cell lymphoma (mycosis fungoides) in a donkey. Our observations suggest that cutaneous T-cell lymphoma should be included in the differential diagnoses of exfoliative dermatitis, even those progressing in a chronic pattern and/or with few or no pruritus.

The upper epidermis of atopic dogs is altered at the functional and structural levels.

BACKGROUND: The pathogenesis of human atopic dermatitis (AD) is complex. Like humans, dogs develop spontaneous AD so this species could be a useful model of study. However, AD has been less characterised in dogs than in humans. OBJECTIVES: To compare the epidermis of normal and spontaneously atopic dogs at the functional and structural levels. ANIMALS: Six healthy and five atopic laboratory Beagle dogs. METHODS AND MATERIALS: Dogs were clinically characterised by general examination, Canine Atopic Dermatitis Extent and Severity Index, 4th iteration (CADESI-04) evaluation and trans-epidermal water loss (TWEL) measurement. Skin biopsies were taken from healthy skin from normal dogs and on nonlesional and lesional skin from atopic dogs. Samples were analysed using transmission electron microscopy (TEM). Cornified envelopes were extracted and examined for their visual aspects (smooth versus ruffled). RESULTS: CADESI-04 and TWEL were significantly higher in atopic dogs. Healthy and nonlesional skin could be distinguished from lesional skin by histopathological evaluation. TEM examination revealed abnormal morphology of the stratum corneum (SC) in atopic skin. The SC compactum corneocyte layer was larger. Thicker and wrinkled corneocytes were more prominent (P = 0.005) in the lesional skin. Similar changes were observed in the nonlesional skin, but less pronounced. The proportion of immature ruffled envelopes was increased in atopic samples (P < 0.05), both from lesional and nonlesional areas. CONCLUSIONS: The morphology of the SC was altered in the lesional and apparently nonlesional skin of spontaneously atopic dogs.

Survey of cytokines on ocular surfaces of atopic dogs by multiplex analysis using two sampling methods - a pilot study.

BACKGROUND: Conjunctivitis in atopic dogs has already been described yet is rarely observed, and likely underdiagnosed in practice. OBJECTIVES: To assay various cytokines in tears and conjunctivae from atopic and normal dogs, and to compare canine atopic dermatitis-associated conjunctivitis with controls. ANIMALS: Ten atopic and ten normal client-owned dogs. METHODS AND MATERIALS: Ocular surfaces were sampled bilaterally in a prospective study, using two different methods. Tear samples were obtained with a sterile swab previously moistened with saline solution (method A). Conjunctival impressions were obtained with a conjunctival impression device (method B). For each sample, the concentrations of a panel of 13 cytokines were measured by multiplex analyses. CADESI-4, pruritus (PS) and conjunctival (CS) scores were determined. RESULTS: Among the measured cytokines, only granulocyte-macrophage colony-stimulating factor (GM-CSF), keratinocyte-derived chemokine (KC)-like and interleukin (IL)-8 were above the limit of quantification in most samples. Absolute amounts of each cytokine were always higher in samples obtained with method A than with method B. GM-CSF amounts were lower in atopic dogs (method A, P=0.02; method B, P=0.0005). KC levels were higher in atopic dogs, yet the differences were not significant. IL-8 amounts were higher in atopic dogs (method A, P=0.0003; method B, P=0.006). CONCLUSIONS AND CLINICAL RELEVANCE: Regardless of the method, these preliminary results suggest an overexpression of IL-8 in conjunctivae and tears of atopic dogs despite subtle conjunctival symptoms. As IL-8 is commonly found in many inflammatory conditions, further studies are needed to determine its specificity in atopic conjunctivitis.

Comparison of three methods for the diagnosis of otoacariasis due to Otodectes cynotis in dogs and cats.

BACKGROUND: The diagnosis of otoacariasis due to Otodectes cynotis is based on the visualization of the parasite, either directly by otoscopy or indirectly after microscopic examination of cerumen collected by several methods. OBJECTIVES: To compare the sensitivity of three techniques: conventional handheld otoscopy, Volkmann's curette sampling and cotton-tipped swabbing. ANIMALS: Five dogs and 12 cats (30 naturally infested ears). METHODS AND MATERIALS: For each case, following otoscopy, the order of examinations (swab or curette) was chosen randomly and the samples were observed on a slide mixed with lactophenol and covered with a coverslip. Parasite detection was noted as positive or negative and parasitic stages were counted separately. RESULTS: The diagnostic sensitivity of otoscopy alone was 67% (positive in 20 of 30 cases), using the curette sampling it was 93% (28/30) and 57% (17/30) for the swabbing. The curette technique had a significantly higher sensitivity than classic ear-swabbing (P = 0.001) or otoscopy alone (P = 0.02). Combining otoscopy and the curette, we obtained a sensitivity of 100% compared to 86% when otoscopy and swabbing were combined. Moreover, the parasite count in the curette samples (average 25 ± 30 SD) was significantly higher than the swab samples (4.5 ± 11) (P < 0.001). CONCLUSIONS AND CLINICAL IMPORTANCE: When suspecting O. cynotis infestation, otoscopic examination should be performed. To confirm the nature of the parasites observed or whenever this examination result is negative, doubtful or cannot be performed, the curettage sampling method for microscopic cerumen examination is recommended.

Reliability of different sets of criteria in diagnosing canine atopic dermatitis applied to a population of 250 dogs seen in a veterinary teaching hospital.

BACKGROUND: Different sets of criteria are available in veterinary dermatology for the diagnosis of canine atopic dermatitis (cAD); there are limited data to assess their reliability. HYPOTHESIS/OBJECTIVES: The aim of this study was to assess the discriminative and predictive ability of four sets in diagnosing cAD in a population of dogs with different skin diseases. ANIMALS: Two hundred and fifty dogs examined in the dermatology unit of a veterinary teaching hospital. METHODS AND MATERIALS: Dogs were diagnosed with cAD, flea infestation (FI), flea bite hypersensitivity (FBH), sarcoptic mange (SM) and other skin diseases (OD). FI, FBH and SM were pooled in one group of dermatoses mimicking cAD (MD). All nonatopic dogs were grouped as a "global population" (GP). Four criteria sets were applied to all dogs. For each set of criteria, sensitivity, specificity, positive and negative predictive values (PPV, NPV), likelihood ratios (LR) and diagnostic odds ratio (DOR) were calculated. RESULTS: When applied to the GP, sensitivities ranged from 54 to 83%, specificities from 68 to 75% and PPVs from 35 to 43%. NPVs ranged from 87 to 94%. LRs and DORs were poor. When applied to MD, sensitivities remained unchanged, specificities and PPV were mildly higher, whereas NPVs were mildly lower and LRs and DORs were comparable. CONCLUSIONS AND CLINICAL IMPORTANCE: This study showed that each set of criteria had a low diagnostic reliability when used alone. The use of more discriminant criteria integrated into a thorough clinical approach excluding MDs should be considered.

Performance and Tolerance of a Protocol for Idiopathic Chronic Greasy Seborrhea in 18 Dogs Using a Shampoo and Mousse Containing Plant Extracts.

The study aimed to evaluate the tolerance, performance and effect on hair lipids and skin hydration of a protocol combining applications of one shampoo and subsequent mousses containing plant extracts (Ophytrium and Seboliance) in dogs with an undiagnosed chronic greasy keratinisation disorder. Six dogs were washed with plain water on day (D)0. Twelve dogs were shampooed on D0 and received eight mousse applications at 48-72 h intervals from D2 to D18. Clinical score (CS), Natural Moisturizing Factors (NMF) and hair lipids (HL) were evaluated on D0, D0 + 4 h, D7, D14 and D24. At baseline, no significant differences were observed in CS, NMF and HL between groups. In the control group, CS and HL remained stable throughout the study while a slight decrease in NMF was observed at D0 + 4 h. CS was significantly reduced in the test group between D0 and D7 (-53%) which reached 91% at D24 (p < 0.05), with no side effects. NMF levels decreased in the test group at D0 + 4 h (-73%, p < 0.0001) and returned to baseline from D14. In conclusion, one shampoo and subsequent mousse applications rapidly and safely improved coat quality in dogs with an undiagnosed keratinisation disorder without affecting NMF and HL contents over the study period.

Multiplex Cytokine Analyses in Ear Canals of Dogs Suggest Involvement of IL-8 Chemokine in Atopic Otitis and Otodectic Mange-Preliminary Results.

Cutaneous cytokines and chemokines are involved in the pathogenesis of human and canine atopic dermatitis. The aim of the present study was to discriminate cytokine expression in the ear canals of atopic dogs with otitis, dogs with non-allergic inflammatory otitis (otodectic mange) and healthy non-atopic dogs. The ear canals of nine atopic dogs suffering from non-infected otitis externa (n = 14 ears), 10 healthy dogs suffering from otodectic mange (n = 20 ears) and 21 healthy controls (39 ears) were swabbed. The concentrations of a panel of 13 cytokines and chemokines on the aural surface were measured by multiplex analyses (Milliplex Canine Cytokine Panel). In addition, Canine Atopic Dermatitis Extent and Severity Index (CADESI)-04 and Otitis Index Score (OTIS3) scores were used to evaluate the overall status of the dogs. The concentration of IL-8 was significantly higher in the ears of atopic dogs and dogs with otodectic mange compared to those of healthy dogs. Significant increases in the levels of IL-10 were also overexpressed in atopic otitis but at lower rates. The concentrations of interleukin(IL)-8 were positively correlated with the OTIS3 hyperplasia score in atopic dogs. Taken together, these results suggest that IL-8 is overexpressed in atopic otitis and otodectic mange and that levels correlate with the otitis severity in atopic dogs.

Clinical and Microbiological Performances and Effects on Lipid and Cytokine Production of a Ceruminolytic Ear Cleaner in Canine Erythemato-Ceruminous Otitis Externa.

Erythemato-ceruminous otitis externa (ECOE) is the most common type of otitis in dogs and is generally associated with bacterial and/or yeast infections. The performance of an ear cleaner was assessed over two weeks in canine ECOE, associated with a mild or moderate secondary infection, in a prospective open-label study. Forty ear canals with ECOE that did not receive any type of aural treatment and were not cleaned for 7 days were included. Pruritus (PS), 0−3 Otitis Index Score (OTIS-3) and 0−4 scale cytology (CYTO) scores were assessed on Day (D) 0, D7 and D14. Concentrations of a panel of 13 cytokines on the ear canal surface and the lipid profile of the exudate were measured on D0 and D14. From D0 to D12 or D13, the dogs' ears were cleaned daily if the secretion score (SEC) was 3/3, every second day if the score was 2/3 and every third day if the score was 1/3. PS, OTIS-3, SEC and CYTO were significantly lower on D7 compared to baseline (−40%, −31%, −36%, −34%, respectively; p < 0.0001). The same parameters decreased further on D14 (−60%, −53%, −61%, −73%, respectively; p < 0.0001) and amounts of interleukin 8 and chemokine KC-like were also reduced compared to baseline (−45%, p < 0.01; −36%, p = 0.3, respectively). The lipid profile was also modified, with a decrease in free lipids and an increase in bound lipids.

Case Report: Positive Outcome of a Suspected Drug-Associated (Immune Mediated) Reaction in a 4-Year-Old Male French Bulldog.

Toxic epidermal necrolysis (TEN) is a rare and severe life-threatening syndrome characterized by apoptosis of keratinocytes resulting in devitalization of the epidermis affecting more than 30% of skin surface. In humans and animals, this condition is mostly triggered by drugs. Identification of the putative agent and its withdrawal are crucial to successful management of a patient with TEN. In this case study, we report the clinical features, histopathological findings and management of a dog with TEN. A 4-year-old intact male French bulldog presented with acute onset of severe lethargy and cutaneous ulcerations on the footpads, scrotum, and hind limbs associated with marked pain. A Stevens-Johnson syndrome/TEN was suspected and drugs, especially beta-lactams, were withdrawn. Histopathology confirmed the diagnosis of epidermal necrosis. Advanced supportive therapy, pain management and skin care led to rapid remission. Early identification and removal of the suspected medication was crucial to improving TEN prognosis in this dog. Antibiotics (penicillin, ampicillin, cephalexin, and sulfonamides) are frequently involved in adverse cutaneous reactions in dogs. Ideal treatment remains elusive is humans and dogs and this disease has a poor prognosis. Supportive care combined with pain management and treatment of the cutaneous ulcerations is essential.

Update on canine filaggrin: a review.

Human filaggrin (FLG) plays a key role in epidermal barrier function, and loss-of-function mutations of its gene are primarily responsible for the development of human atopic dermatitis (AD). FLG expression is also reduced in the epidermis of atopic patients, due to the transcriptional effect of Th2 type cytokines. Canine atopic dermatitis (CAD) is a prevalent skin disease that shares many clinical and pathogenic features with its human homologue. The aim of this review is discuss current knowledge on canine filaggrin (Flg) in both healthy and atopic dogs, as compared to the human protein. Although the molecular structures of the two proteins, as deduced from the sequences of their gene, are different, their sites of expression and their proteolytic processing in the normal epidermis are similar. Concerning the expression of Flg in CAD, conflicting results have been published at the mRNA level and little accurate information is available at the protein level. It derives from a large precursor, named profilaggrin (proFLG), formed by several FLG units and stored in keratohyalin granules of the stratum granulosum. Canine and human proFLG sequences display little amino acid similarity (33% as shown using the Basic Local Alignment Search Tool (BLAST)) except at the level of the S100 homologous part of the N-terminus (75%). Genetic studies in the dog are at an early stage and are limited by the variety of breeds and the small number of cases included. Many questions remain unanswered about the involvement of Flg in CAD pathogenesis.

Clinical and histological recovery of non-thymoma-associated exfoliative dermatitis in a cat treated with ciclosporin A.

CASE SUMMARY: A 6-year-old neutered female European Shorthair cat was referred for chronic, moderately pruritic, alopecic and exfoliative dermatosis that was unresponsive to antiparasitic, antibiotic or steroidal anti-inflammatory drugs. The cat presented with truncular alopecia and numerous whitish adherent scales covering the whole body. Differential diagnoses included sebaceous adenitis, dermatophytosis, demodicosis, exfoliative dermatitis associated or not with thymoma, drug reaction, feline immunodeficiency virus- or feline leukaemia virus-associated dermatoses, epitheliotropic T-cell lymphoma, and Malassezia yeasts and/or bacterial overgrowth. Blood tests were within normal limits and the retrovirus tests were negative. Skin scrapings, fungal culture, coat brushing and skin cytology were negative for parasitic or microbial elements. Radiographs showed no signs of a thymic mass. Histological examination of skin biopsies revealed marked orthokeratotic hyperkeratosis, lymphocytic (CD3+) interface dermatitis and mural folliculitis with absence of sebaceous glands and occasional apoptotic cells in different epidermal layers. Clinical and histological findings were consistent with non-thymoma-associated exfoliative dermatitis syndrome. Ciclosporin A (7 mg/kg) was administered once daily. A dramatic improvement was observed after 3 weeks. Ciclosporin A intake was then progressively spaced out as the clinical signs diminished. Skin biopsies revealed resolution of hyperkeratosis, disappearance of the inflammatory infiltrate and recovery of the sebaceous glands. RELEVANCE AND NOVEL INFORMATION: T-cell infiltration with signs of epidermal cytotoxicity, in the absence of infectious agents or neoplastic process, suggests an immune-mediated process, and ciclosporin A, a calcineurin inhibitor, would be the drug of choice. This is the first report showing resolution of both the clinical and histological signs of non-thymoma-associated exfoliative dermatitis.

Multinodular Malignant Cutaneous Mast Cell Tumor in a Horse With Generalized Pruritus and Reactive Fibrosis: A Case Report.

Mast cell tumor (MCT) has long been considered as an uncommon neoplasm in horses. Cytological and behavioral evidence of its malignancy is usually lacking, and only a few reports have described MCT displaying malignant behavior. An 18-year-old Friesian stallion presented with a one-year history of intermittent and progressive skin lesions on the left forelimb associated with intense, generalized pruritus and apathy temporarily responsive to glucocorticoids and antibiotics. The horse was alert and responsive with poor body condition and marked generalized pruritus. The left forelimb was markedly enlarged and surrounded by numerous firm 2- to 20-cm masses that were ulcerated and focally necrotic. A 7-cm round firm mass was observed on the left dorsal neck. Dermatological examination revealed generalized moth-eaten alopecia and scaling with erosions and ulcers secondary to pruritus. A direct skin smear from the affected leg showed severe eosinophilic inflammation and neutrophilic inflammation with pleomorphic bacteria. Histopathology of the skin and biopsies of the underlying tissues revealed an abundant population of atypical mast cells consistent with a malignant MCT. The horse was euthanized and necropsy revealed a marked fibrous reaction on longitudinal sections of the affected limb, and the tumor could be detected on only a few histological slides. Diagnosis of equine MCT can be challenging because of the massive accompanying fibrous reaction. Mast cell tumor should be suspected in the presence of eosinophilic infiltration of the affected tissue and in cases of generalized pruritus not attributable to other causes.

Treatment of Harvest Mite Infestation in Dogs Using a Permethrin 54.5% and Fipronil 6.1% (Effitix®) Topical Spot-On Formulation.

Background-The study aims to assess the efficacy of a permethrin 54.5%-fipronil 6.1%-based spot-on solution in dogs naturally infested with Neotrombicula in an open-label controlled study. Methods-Ten naturally infested dogs received one drop per affected site on day (D) 0, and on D14, the rest of the pipette was applied on the skin between the shoulders. Five dogs served as sentinels. Parasite score (0-3), skin lesions (0-4), and investigator pruritus scale (0-4) were assessed on D0, D1, D14, and D28. Results-No treated dogs developed adverse reactions. Parasite score of sentinel dogs was maintained between 1.8 (D0, D1, and D28) and 2.2 (D14). In treated dogs, D0 parasite score was 2.4. It was significantly reduced from D1 (0.5; p < 0.002) to D28 (0.1; p < 0.002). The lesion score was 2.9 on D0 and D1; it was significantly reduced on D14 (0.6; p < 0.002) and D28 (0.1; p < 0.002). Similarly, investigator pruritus scale (D0, 2.2) scores significantly decreased on D14 (0.4; p < 0.004) and D28 (0.2; p < 0.002). Conclusions-The combination permethrin-fipronil appears to be well-tolerated, rapidly and durably effective in the control of localized canine harvest mite infestation.