Giordana Zanna

A novel unstable duplication upstream of HAS2 predisposes to a breed-defining skin phenotype and a periodic fever syndrome in Chinese Shar-Pei dogs.

Hereditary periodic fever syndromes are characterized by recurrent episodes of fever and inflammation with no known pathogenic or autoimmune cause. In humans, several genes have been implicated in this group of diseases, but the majority of cases remain unexplained. A similar periodic fever syndrome is relatively frequent in the Chinese Shar-Pei breed of dogs. In the western world, Shar-Pei have been strongly selected for a distinctive thick and heavily folded skin. In this study, a mutation affecting both these traits was identified. Using genome-wide SNP analysis of Shar-Pei and other breeds, the strongest signal of a breed-specific selective sweep was located on chromosome 13. The same region also harbored the strongest genome-wide association (GWA) signal for susceptibility to the periodic fever syndrome (praw = 2.3×10−6, pgenome = 0.01). Dense targeted resequencing revealed two partially overlapping duplications, 14.3 Kb and 16.1 Kb in size, unique to Shar-Pei and upstream of the Hyaluronic Acid Synthase 2 (HAS2) gene. HAS2 encodes the rate-limiting enzyme synthesizing hyaluronan (HA), a major component of the skin. HA is up-regulated and accumulates in the thickened skin of Shar-Pei. A high copy number of the 16.1 Kb duplication was associated with an increased expression of HAS2 as well as the periodic fever syndrome (p<0.0001). When fragmented, HA can act as a trigger of the innate immune system and stimulate sterile fever and inflammation. The strong selection for the skin phenotype therefore appears to enrich for a pleiotropic mutation predisposing these dogs to a periodic fever syndrome. The identification of HA as a major risk factor for this canine disease raises the potential of this glycosaminoglycan as a risk factor for human periodic fevers and as an important driver of chronic inflammation.

Long term follow-up of dogs diagnosed with leishmaniosis (clinical stage II) and treated with meglumine antimoniate and allopurinol

Twenty-three dogs with a diagnosis of leishmaniosis (clinical stage II) were treated with meglumine antimoniate and allopurinol and were followed up for 2-9 years. The treatment showed efficacy and the clinical condition of the dogs improved markedly in the first 3 months of treatment. Anti-Leishmania antibody titres declined slowly although most dogs remained seropositive 1 year after beginning treatment. Inter-individual variability in the evolution of the titres was very high. The dogs presented with three types of complications during the follow-up period. (1) Three dogs experienced relapses characterized by clinical signs, high anti-Leishmania titres and high parasitaemia. (2) Eight dogs presented immune-mediated lesions, such as uveitis, arthritis and cutaneous vasculitis; in all of these cases, the dogs had high titres of anti-Leishmania antibodies at diagnosis and during follow-up. (3) Three dogs presented xanthine urolithiasis most likely due to the allopurinol treatment. In one case the xanthine uroliths led to hydronephrosis and nephrectomy. The study demonstrated a long survival for dogs with leishmaniosis treated with the combination of meglumine antimoniate and allopurinol. Clinicians should pay special attention to the appearance of immune-mediated lesions, especially in dogs with sustained high antibody titres, and to urolithiasis.

Cutaneous mucinosis in shar-pei dogs is due to hyaluronic acid deposition and is associated with high levels of hyaluronic acid in serum

Cutaneous mucinosis affects primarily shar-pei dogs. Hyaluronic acid (HA) is considered to be the main component of mucin and CD44 is the major cell surface receptor of HA, necessary for its uptake and catabolism. The aims of this study were to identify the composition of the mucin in cutaneous mucinosis of shar-pei dogs, investigate the correlation between the deposition of HA and the expression of CD44, and determine whether shar-pei dogs with cutaneous mucinosis presented with elevated levels of serum HA. In skin biopsies, the mucinous material was stained intensely with Alcian blue and bound strongly by the hyaluronan-binding protein. No correlation was found between the degree of HA deposition in the dermis and the expression of CD44 in the skin of shar-pei dogs affected or unaffected by cutaneous mucinosis. A clear positive correlation was found between the existence of clinical mucinosis and the serum HA concentration. In control dogs, serum HA ranged from 155.53 to 301.62 microg L(-1) in shar-pei dogs; without mucinosis it ranged from 106.72 to 1251.76 microg L(-1) and in shar-pei dogs with severe mucinosis it ranged between 843.51 to 2330.03 microg L(-1). Altogether, the results reported here suggest that mucinosis of shar-pei dogs is probably the consequence of a genetic defect in the metabolism of HA.

Hereditary cutaneous mucinosis in shar pei dogs is associated with increased hyaluronan synthase-2 mRNA transcription by cultured dermal fibroblasts.

Shar pei dogs are known for the distinctive feature of thick, wrinkled skin as a consequence of high dermal mucin content. Excessive dermal deposition of mucinous substance leading to severe skin folding, and/or to the more severe vesicular form characterized by dermal vesicles or bullae, is highly prevalent in this breed and is known as idiopathic mucinosis. Hyaluronic acid (HA) is the main component that accumulates in the dermis, and high levels of HA have also been detected in the serum of shar pei dogs. In this study, the cellular and molecular mechanisms underlying cutaneous mucinosis of shar pei dogs were investigated. Thirteen shar pei dogs and four control dogs of other breeds were included. In primary dermal fibroblast cultures, transcription of the family of hyaluronan synthases (HAS) involved in HA synthesis, and of hyaluronidases (HYAL) involved in HA degradation, were studied by reverse transcriptase polymerase chain reaction. The location of HA in cell cultures was studied by immunofluorescence and confocal laser microscopy. Dermal fibroblasts transcribed HAS2, HAS3, HYAL1 and HYAL2, but no amplification for HAS1 was found. A higher transcription of HAS2 was demonstrated in shar pei dogs compared with control dogs. By confocal microscopy, HA was detected as a more diffuse and intense network-like pattern of green fluorescence in the fibroblast cells of shar pei dogs in comparison with control dogs. Together, these results provide additional evidence that hereditary cutaneous mucinosis in shar pei dogs may be a consequence of over-transcription or increased activity of HAS2.

Increased HAS2‐driven hyaluronic acid synthesis in shar‐pei dogs with hereditary cutaneous hyaluronosis (mucinosis)

The Chinese shar-pei dog is known for its distinctive feature of wrinkled and thickened skin, defined as primary or hereditary cutaneous mucinosis. In a recent report, we identified the mucinous material deposited in the shar-pei skin as the polysaccharide hyaluronan (HA). In the present work, the molecular and cellular mechanisms underlying this phenotype have been identified in dermal fibroblasts isolated from shar-pei dogs. The production of HA, which appeared to be mainly associated with cell membrane protrusions and also intracellular, was higher in shar-pei fibroblasts than in control cells. The HA accumulation is related to a higher mRNA expression of the isoform HAS2 of the HA-synthesizing enzyme family, hyaluronan synthases (HAS). The higher expression of HAS2 in shar-pei fibroblasts was confirmed at the protein level. The other HAS isoenzymes, HAS1 and HAS3, and the HA-degrading enzymes, Hyal1 and Hyal2, were not differentially expressed in shar-pei fibroblasts compared with cells from control dogs. Fibroblasts from shar-pei dogs and from control dogs are morphologically different as observed by transmission electron microscopy. Scanning electron microscopy revealed a large number of cellular protrusions with associated globular deposits. Electron microscopy after labelling with biotinylated HA-binding protein confirmed an increased HA content in shar-pei fibroblasts, which could be localized in several subcellular structures. The authors propose the name hereditary cutaneous hyaluronosis (HCH) for affected dogs, because it better defines the cutaneous mucinosis of shar-pei dogs.

Dermoscopic features in 12 cats with dermatophytosis and in 12 cats with self-induced alopecia due to other causes: An observational descriptive study

BACKGROUND Dermoscopy is a noninvasive technique allowing rapid magnified in vivo observation of the skin and structures that lie beneath the skin surface. Various congenital and acquired hair shaft abnormalities may also be evaluated by dermoscopy. Additionally, characteristic features of Microsporum canis-induced tinea capitis and trichotillomania in humans have been reported. OBJECTIVES To describe the dermoscopic findings observed in cats with patchy alopecia due to M. canis infection and in cats with self-inflicted hair loss. ANIMALS Twenty-four client-owned cats presented at a veterinary referral practice. METHODS Dermoscopy was performed with a hand-held nonpolarized light dermoscope at 10-fold magnification. The glass plate of the dermoscope was applied gently to the lesions and no sedation was required. RESULTS Twelve cats were diagnosed with dermatophytosis and 12 with self-induced alopecia due to other causes. At 10-fold magnification, the most characteristic findings observed in circumscribed lesions of cats with dermatophytosis were opaque, slightly curved, broken hairs of a homogeneous thickness (comma-like structures) and a variable amount of brown-to-yellow greasy scales. In cats with self-induced alopecia, multiple hairs with a normal shaft cleanly broken at different lengths, short tufts of hairs broken at an equal level and hook-like and coiled hairs were observed. CONCLUSIONS AND CLINICAL IMPORTANCE This observational descriptive study suggests that dermoscopy may represent a helpful noninvasive in vivo technique in the differential diagnosis of patchy alopecia in cats.

Dermoscopic evaluation of skin in healthy cats

BACKGROUND Dermoscopy is a diagnostic tool that can reveal morphological structures not visible upon clinical examination. HYPOTHESIS/OBJECTIVES To assess the usefulness and applicability of dermoscopy for the examination of healthy cat skin. ANIMALS Twenty-one domestic short-haired cats from a feline rescue association. METHODS Four regions (head, dorsal neck, sacral and abdominal regions) were examined with both a contact hand-held nonpolarized light dermoscope at 10-fold magnification and a videodermoscope at 70-fold magnification. Findings were assessed using histological analysis of skin samples cut both longitudinally and transversely, set as the gold standard. RESULTS With a hand-held dermoscope at 10-fold magnification, thick, straight primary hairs surrounded by multiple secondary hairs were observed. With a videodermoscope at 70-fold magnification, hair shaft thickness was measured and the follicular openings and arrangement of vessels were clearly observed. Correspondence was observed between dermoscopic and histological results. CONCLUSIONS AND CLINICAL IMPORTANCE Dermoscopy represents a valid noninvasive and reproducible technique that could be helpful in clinical examination.

Evaluation of ultrasonography for measurement of skin thickness in Shar-Peis

OBJECTIVE To determine whether high-frequency diagnostic ultrasonography is useful for assessment of skin thickness in Shar-Peis. ANIMALS 10 healthy Shar-Peis and 10 healthy Beagles used as controls. PROCEDURES Ultrasonographic examination of the skin was performed on 4 cutaneous sites by use of a 13-MHz linear-array transducer, and the mean of 3 measurements was calculated. Ultrasonography results were compared with histologic findings of skin specimens stained with H&E, Alcian blue at a pH of 2.5, and Masson trichrome stains, with histometric measurements of skin thickness made by use of a microscope, and with measurements of skin thickness made by use of a plicometer. Ultrasonograpy results were also compared via age and sex of selected animals. RESULTS A clear correlation was detected between ultrasonography results and results of histologic and histometric analysis in both groups. In Shar-Peis, no correlation was found between ultrasonography results and age and sex, whereas in Beagles, a weak positive correlation was found only between skin thickness in dorsal cervical and frontal (on the rostral margins of the supraorbital processes) regions and age. A positive overall correlation was found in Shar-Peis between measurements made via ultrasonography and plicometery. CONCLUSIONS AND CLINICAL RELEVANCE Ultrasonography was a useful tool to assess skin thickness, and in Shar-Peis, it might be considered a valid alternative to invasive methods such as histologic examination to objectively estimate the severity of hereditary cutaneous hyaluronosis.

Rifampicin treatment of canine pyoderma due to multidrug-resistant meticillin-resistant staphylococci: a retrospective study of 32 cases

BACKGROUND Rifampicin has received increased interest in veterinary dermatology because of its activity against multidrug-resistant meticillin-resistant staphylococci (MRS). There is limited knowledge about the efficacy and safety of rifampicin in dogs. HYPOTHESIS/OBJECTIVE To provide information on response to treatment and adverse effects in dogs treated with rifampicin for multidrug-resistant MRS pyoderma. ANIMALS Thirty two dogs treated with rifampicin for rifampicin-susceptible multidrug-resistant MRS pyoderma. METHODS Retrospective review of medical records, including alanine aminotransferase (ALT) and alkaline phosphatase (ALP) serum activity levels and total bilirubin concentrations, obtained before and throughout the treatment, was performed. RESULTS Oral rifampicin as sole systemic antimicrobial therapy (median dose 5 mg/kg twice daily) was effective in 71.88% of cases. Topical antimicrobials were used in most cases. Median duration of rifampicin treatment was five weeks for superficial pyoderma and four weeks for deep pyoderma. Gastrointestinal signs were reported in 15% of treated dogs. Statistically significant increases of ALT (P = 0.045) and ALP (P = 0.0002) values after 3-4 weeks of treatment was observed. The median increase was equal to 0.3 and ×1.5 the upper limit of the reference ranges for ALT and ALP, respectively. CONCLUSIONS/CLINICAL IMPORTANCE Oral rifampicin combined with topical antimicrobials can be considered an effective therapeutic option for canine superficial and deep pyoderma caused by rifampicin-susceptible multidrug-resistant MRS. Liver enzyme induction might be the most important cause of ALT and ALP increase associated with rifampicin therapy in dogs.