Unveiling the Role of Endoplasmic Reticulum Stress Pathways in Canine Demodicosis.

Canine demodicosis is a prevalent skin disease caused by overpopulation of a commensal species of Demodex mite, yet its precise cause remains unknown. Research suggests that T-cell exhaustion, increased immunosuppressive cytokines, induction of regulatory T cells and increased expression of immune checkpoint inhibitors may contribute to its pathogenesis. This study aimed to gain a deeper understanding of the molecular changes occurring in canine demodicosis using mass spectrometry and pathway enrichment analysis. The results indicate that endoplasmic reticulum stress promotes canine demodicosis through regulation of three linked signalling pathways: eIF2, mTOR, and eIF4 and p70S6K. These pathways are involved in the modulation of Toll-like receptors, most notably TLR2, and have been shown to play a role in the pathogenesis of skin diseases in both dogs and humans. Moreover, these pathways are also implicated in the promotion of immunosuppressive M2 phenotype macrophages. Immunohistochemical analysis, utilising common markers of dendritic cells and macrophages, verified the presence of M2 macrophages in canine demodicosis. The proteomic analysis also identified immunological disease, organismal injury and abnormalities and inflammatory response as the most significant underlying diseases and disorders associated with canine demodicosis. This study demonstrates that Demodex mites, through ER stress, unfolded protein response and M2 macrophages contribute to an immunosuppressive microenvironment, thereby assisting in their proliferation.

Pathology of "double scale" skin defect in farmed American alligators (Alligator mississippiensis) and the possible association with hepatic fibrosis.

"Double scale" is a poorly characterized skin defect of crocodilians that drastically reduces the economic value of crocodilian skin. This study investigated the morphology and pathogenesis of double scale in a ranching farm of American alligators (Alligator mississippiensis). We compared the histopathology of skin and selected organs (liver, lung, kidney, heart, spleen, intestine, and brain) of alligators with double scale against healthy control animals, together with serum and liver vitamin and mineral levels. Skin affected with double scale had statistically significant hyperkeratosis, epidermal atrophy, and increased basal cell degeneration compared with control alligators (P < .0001). Interestingly, all alligators with double scale had varying degrees of hepatic fibrosis. Feed analysis showed that alligators that had double scale and hepatic fibrosis had prolonged dietary exposure to high levels of vitamin A, iron, and copper. Serum analysis indicated that levels of zinc (p < .0001), copper (P < .05), and vitamin E (P < .002) were significantly lower in alligators with hepatic fibrosis and double scale compared with controls. Finally, immunohistochemical analysis of skin with double scale showed a marked reduction in immunolabeling with the zinc-binding protein metallothionein. These results suggest that zinc deficiency, in combination with other micronutrient anomalies, may play a role in the pathogenesis of double scale in alligators with liver fibrosis.

Gene expression analysis of Canine Demodicosis; A milieu promoting immune tolerance.

Canine demodicosis is a common skin disease seen in companion animal practice that results from an overpopulation of the commensal Demodex mite species. Common predisposing factors to the development of canine demodicosis include immunosuppressive diseases, such as neoplasia and hypothyroidism, and administration of immunosuppressive therapies, such as corticosteroids. Despite this, the pathogenesis of development of canine demodicosis remains unclear. Previous studies have implicated a role for increased expression of toll like receptor 2 (TLR2), increased production of interleukin (IL)-10) and T cell exhaustion. Here, we investigate gene expression of formalin fixed paraffin embedded skin samples from twelve cases of canine demodicosis in comparison to twelve healthy controls, using a 770 gene panel (NanoString Canine IO Panel). Results show an increase in the T cell population, specifically Th1 and Treg cells in dogs with demodicosis. In addition, while there is an upregulation of immunosuppressive cytokines such as IL-10 and IL-13, there is also an upregulation of immune check point molecules including PD-1/PD-L1 and CTLA-4. These findings suggest that Demodex spp. mites are modulating the host immune system to their advantage through upregulation of several immune tolerance promoting pathways.

A retrospective study of cases of canine demodicosis submitted to a commercial diagnostic laboratory servicing the United Kingdom and Ireland (2017-2018) part 2; Aerobic culture and antimicrobial susceptibility results.

Clinical diagnostic reports from 508 cases of canine demodicosis diagnosed either by histological or skin scraping analysis from a United Kingdom Accreditation Service (UKAS) accredited veterinary diagnostic laboratory servicing the United Kingdom (UK) and Ireland were evaluated. Of the 508 cases, 284 had skin swabs submitted for culture on the same day the skin biopsy and/or skin scraping were obtained. Dogs with juvenile-onset (JO) demodicosis represented 57.4% of these cases, whilst adult-onset (AO) cases comprised 42.6%. The data revealed that overgrowth of pathogenic bacteria was more common in AO demodicosis cases (75.2%) in comparison to the JO cases (57%). Adult-onset cases also had increased involvement of bacteria belonging to multiple genera and/or yeast (28.9%) in comparison to JO cases (18.4%). Pruritus was significantly associated with an overgrowth of Staphylococcus pseudintermedius (p < 0.001). Resistance to one or more antimicrobial classes was noted in S. pseudintermedius isolates from 56.3% of JO cases with 10.3% of these cases being classified as Multi-Drug Resistant (MDR). Similarly, 51.9% of S. pseudintermedius isolates from the AO cases were noted to be resistant to one or more antimicrobial class with 8.6% of these cases being considered MDR. Cephalosporins were the most frequently administered antimicrobial class noted in submission histories, followed by the penicillin and fluoroquinolone classes. Whilst our findings reveal a high prevalence of concurrent overgrowth of pathogenic bacteria warranting therapeutic intervention in canine demodicosis, the presence of resistance within isolates highlights the need for prudent selection and targeted use of antimicrobial therapy that encompass the key principles of antimicrobial stewardship.

A retrospective study of cases of canine demodicosis submitted to a commercial diagnostic laboratory servicing the United Kingdom and Ireland (2017-2018): Part 1 - Signalment, lesion distribution, treatments, and concurrent diseases.

Canine demodicosis, due to an overpopulation of Demodex spp. mites, remains one of the most common dermatological diseases encountered in small animal practice. The aims of this study were to interrogate submitted histories and diagnostic report results from a large cohort of dogs (n = 508) diagnosed with demodicosis either through histological analysis or the finding of Demodex spp. mites on skin scrapings by a UKAS accredited commercial laboratory servicing the United Kingdom (UK) and Ireland in the years 2017 and 2018. The main findings revealed that short-coated breeds were more likely to develop juvenile-onset (JO) demodicosis, whereas medium- and long-coated breeds were more likely to develop adult-onset (AO) disease. Pododemodicosis was reported more commonly in adult, long-coated breeds. Skin scrapings were positive in only 83.3% of samples that had a corresponding positive biopsy result; this finding highlights the necessity to perform further diagnostic tests if demodicosis remains clinically suspected despite a negative skin scraping result. Concurrent underlying diseases, potentially associated with immunosuppression, were reported in 42/221 (19%) of dogs with AO demodicosis. Serum allergy and Sarcoptes ELISA assays were positive in individual animals in both the JO and AO groups; the clinical significance of these latter findings requires careful interpretation in dogs with confirmed demodicosis.

House dust mite-treated PAR2 over-expressor mouse: A novel model of atopic dermatitis.

BACKGROUND: Atopic dermatitis (AD) is a complex skin disease involving causative effects from both intrinsic and extrinsic sources. Murine models of the disease often fall short in one of these components and, as a result, do not fully encapsulate these disease mechanisms. OBJECTIVE: We aimed to determine whether the protease-activated receptor 2 over-expressor mouse (PAR2OE) with topical house dust mite (HDM) application is a more comprehensive and clinically representative AD model. METHODS: Following HDM extract application to PAR2OE mice and controls, AD clinical scoring, itching behaviour, skin morphology and structure, barrier function, immune cell infiltration and inflammatory markers were assessed. Skin morphology was analysed using haematoxylin and eosin staining, and barrier function was assessed by transepidermal water loss measurements. Immune infiltrate was characterised by histological and immunofluorescence staining. Finally, an assessment of AD-related gene expression was performed using quantitative RT-PCR. RESULTS: PAR2OE mice treated with HDM displays all the characteristic clinical symptoms including erythema, dryness and oedema, skin morphology, itch and inflammation typically seen in patients with AD. There is a significant influx of mast cells (P < .01) and eosinophils (P < .0001) into the dermis of these mice. Furthermore, the PAR2OE + HDM mice exhibit similar expression patterns of key differentially expressed genes as seen in human AD. CONCLUSION: The PAR2OE + HDM mouse presents with a classic AD pathophysiology and is a valuable model in terms of reproducibility and overall disease representation to study the condition and potential therapeutic approaches.