PLENARY SESSION ABSTRACTS Theme: IMMUNITY and AUTOIMMUNITY

PLENARY SESSION ABSTRACTS Theme: IMMUNITY AND AUTOIMMUNITY

State-of-the-Art Address Supporting Review What’s new in autoimmune blistering diseases? Epithelial, immune cell and microbial cross- D. F. MURRELL talk in homeostasis and atopic dermatitis Department of Dermatology, St George Hospital, and T. KOBAYASHI UNSW Faculty of Medicine, Sydney, New South Wales, Laboratory for Innate Immune Systems, RIKEN Center Australia for Integrative Medical Sciences (IMS), Yokohama, There are several blistering diseases which occur natu- Japan rally in other species as well as in humans; for example, Skin is a complex and dynamic ecosystem, wherein the pemphigus occurs naturally in dogs and horses and the epithelial cells, immune cells and microbiota engage in inherited blistering disease, epidermolysis bullosa, also active dialogues and maintain barrier integrity and occurs in dogs. Several new validated scoring systems functional immunity. Alterations of the peaceful coexis- to measure the severity of autoimmune blistering dis- tence with the resident microbiota, referred to as dys- ease (AIBD) have been developed which assist in biosis, lead to dysregulation of host immunity. It has demonstrating efficacy of new treatments, such as the been long debated whether the dysbiosis in the skin of Pemphigus Disease Area Index (PDAI) for pemphigus atopic dermatitis is merely a consequence of chronic and Bullous Pemphigoid Disease Area Index (BPDAI) skin inflammation or whether it is actively involved in for pemphigoid. Pemphigus is due to autoantibodies to driving skin inflammation. Microbiome analysis by 16S desmogleins 1 and 3 in human pemphigus foliaceus and rRNA sequencing in humans and dogs with atopic der- vulgaris and desmocollin1 in canine pemphigus foli- matitis showed the shifts in microbial diversity repre- aceus, generated by the late onset activation of the sented by increased proportion of Staphylococcus spp. adaptive immune response. Paraneoplastic pemphigus, Direct evidence has been found in mice deficient in a in which there are also antibodies to plakin proteins in disintegrin and metalloproteinase 17 (ADAM17), which the skin and mucosae, also has occurred in canines. spontaneously developed eczematous dermatitis with The B-cell CD-20 inhibitor, Rituximab, with short- dysbiosis that was predominated by S. aureus and course steroids has been shown to be more effective Corynebacterium spp. Targeting the dysbiotic flora with and safer than traditional high-dose steroids alone in a an antibiotic cocktail reversed dysbiosis and extin- large human pemphigus RCT. guished eczematous inflammation, suggesting crucial Pemphigus also involves the innate immune response, roles of dysbiotic flora during atopic inflammation. with induction of interleukin (IL)-1 and IL-6. A canine Symbiotic relationships between the host and micro- version of the PDAI has been developed and used in a biota must be constitutively maintained. Detailed mech- pivotal study of the first Bruton Tyrosine Kinase Inhi- anisms on how host immunity regulates commensal bitor (BTKI), which can inhibit both the innate and bacteria in the steady state are now being revealed. adaptive immune response, to be used in an AIBD, in Skin harbours a myriad of tissue-resident immune cells dogs, as solo open-label treatment. Two BTKIs given which include both innate and adaptive immune cells. orally once daily demonstrated excellent efficacy and Recent studies have highlighted a fundamental role of safety, as proof-of-concept for trials in patients with innate lymphoid cells (ILCs) in the maintenance of bar- pemphigus. Open-label Phase 2 studies in pemphigus rier functions and tissue homeostasis. A study in mice have shown rapid responses with rilzabrutinib and the revealed that ILCs and T lymphocytes differentially reg- phase 3 PEGASUS trial is recruiting. CAR-T cell trials ulate skin microbiota. In particular, epidermal ILCs reg- in pemphigus are also ongoing. Bullous pemphigoid ulate production of antimicrobial lipids from sebaceous (BP) is increasing in incidence, particularly in patients glands to maintain homeostatic balance of skin micro- over 70 years. Case control studies have shown that biota. Cross-talk between epithelial cells and immune prior neurological damage, particularly multiple sclero- cells determines immunological tones in the barrier tis- sis, and also dementia and Parkinson disease, increase sue. ILCs directly respond to tissue-derived signals and the risk. Multiple medications increase the risk of trig- play an essential role in barrier immunity. Epithelial cells gering BP, particularly gliptins used to treat diabetes produce alarmins such as thymic stromal lymphopoietin mellitus. Eotaxin 1, IL-17 and IL-4/13 are pathways (TSLP), interleukin (IL)-33 and IL-25, all of which acti- which are increased in BP and trials of new therapies vate group 2 ILCs (ILC2s), which produce type 2 cytoki- targeting these pathways are underway. Epidermolysis nes such as IL-5 and IL-13, and boost type 2 immune bullosa acquisita (EBA) develops due to autoantibodies reactions against parasitic infections, allergens, mites and to collagen VII, the anchoring fibril protein present in toxins. Dysregulation of epithelial–ILC cross-talk results skin and stratified mucosae. Inflammatory forms of in allergic inflammation. Single cell RNA-sequencing EBA have been reproduced in mice and respond to analysis uncovered unique characteristics of skin ILCs complement inhibitors and inhibition of the Fli-1 path- and the regulatory mechanisms in the context of atopic way, which is increased in the EBA mouse and in inflammation. Understanding active interactions between hereditary dystrophic EB, a disease with gene defects in the host epithelial and resident immune cells and micro- collagen VII. biota provides a foundation of novel therapeutic strate- Source of funding: Self-funded. gies of inflammatory skin diseases. Conflict of interest: None declared. Source of funding: Self-funded. Conflict of interest: None declared.

State-of-the-Art Address syndrome, and that different genetic alterations play a A world of inflammation: ecological solutions role in certain breeds and geographical locations and for people, place and planet not in others. Disease development is modulated by the interaction between skin barrier alterations, lifestyle S. L. PRESCOTT conditions, microbiome and immunological responses. School of Paediatrics and Child Health, University of As is the case in people, dogs living on a farm with Western Australia, Perth, Western Australia, Australia increased exposure to the outdoors and other dogs have The ecology of the early environment determines life- decreased risk of development of disease. This likely long health, including microbial diversity, nutrition, results from a modulation of immunological response nature, social interactions and the wider “exposome”. by increased exposure to beneficial bacteria and Almost all of these factors impacting immunity had increased biodiversity. Skin barrier impairment plays a have implications for all aspects of health and reliance. role in promoting cutaneous dysbiosis, increased aller- In particular, the emergence of “microbiome science” gen penetration and excessive T helper 2 (Th2) provides new evidence of vital relationships between response. It is under investigation whether primary biodiversity and health at every level. Allergy was the defects exist in dogs, as reported in people, or whether first discipline to link adverse changes in early life ecol- alterations are purely secondary to inflammation. Doc- ogy with the epidemic of immune disease – with much umented abnormalities range from ultrastructural (e.g. wider implications for other systems. New perspectives disorganized lipid lamellae) to chemical (e.g. decreased of ecological interdependence connect personal and ceramide levels) and functional (e.g. increased transepi- planetary health; the human health crisis cannot be sep- dermal water loss). These alterations are present in clin- arated from the social, political and economic “ecosys- ically normal atopic skin and are intensified in lesional tems” otherwise driving “dysbiosis” (life in distress) at atopic skin. It is important to stress that clinically nor- every level. Changes in macroscale ecology – of food mal atopic skin is not the same as normal skin and that systems, lifestyle behaviours, socioeconomic disadvan- low-grade inflammation is present in atopic skin, thus tage and environmental degradation – all impact the making possible that these changes are secondary. microbial systems sitting at the foundations of all Future studies should include control groups with ecosystems. In particular, changes in the function and inflammatory diseases that are not AD (besides healthy composition of the human-associated microbiome has controls) to distinguish between changes that are simply been implicated in the mounting global burden of non- consequences of inflammation and those that are speci- communicable diseases (NCDs), exacerbating inflam- fic signatures of atopic disease. Decreased transepithe- mation and metabolic dysregulation through multiple lial electrical resistance of monolayers of atopic pathways along lifespans. This underscores the need for keratinocytes in culture suggests a role of tight junctions ecological approaches aimed at restoring symbiosis, bal- or other proteins involved in cell–cell connection. Pro- ance and mutualism. Although there is promise with teins important for keratinization such as filaggrin(s) supplement-based strategies (e.g. probiotics, prebiotics), have been investigated. In genetic studies, it was found it is essential to focus on upstream factors implicated in that filaggrin single nucleotide polymorphism (SNP) dysbiosis, including the health of wider environments, may play a role in some breeds and not others. Of lifestyle, nature relatedness, and the social policies and interest is a SNP in the gene encoding for thymic stro- practices which can facilitate or inhibit “dysbiotic mal lymphopoietin (TSLP) receptor, which was drift”. This underscores the necessity for ambitious detected in all tested breeds. TSLP is an important link integrative approaches which not only define these between skin barrier damage and modulation of Th2 interconnections, but also capitalize on them to create response. Interestingly, staphylococcal wall components novel, collaborative and mutualistic solutions to our can induce transcription of TSLP. Documented defects vast interdependent global challenges. of extrusion of antimicrobial peptides potentially can Source of funding: Self-funded. explain reduced killing activity in atopic patients and Conflict of interest: None declared. the important role of Staphylococcus in promoting flares of AD. Decreased biodiversity and increased Supporting Review staphylococcal colonization is present in atopic skin Advances in our understanding of canine atopic and aggravated during active flares. A dysregulated immune response in cAD includes increased Th2, Th17 dermatitis and CD4+ CD25+ regulatory T cells. Interleukin (IL)- R. MARSELLA 31 has been identified as an important mediator of pru- Department of Small Animal Clinical Sciences, College ritus and as a possible biomarker for disease severity. of Veterinary Medicine, University of Florida, As results have varied among studies, more work is Gainesville, FL, USA needed to evaluate IL-31 expression in various pheno- Much progress has been made in our understanding types of cAD. In summary, our understanding has canine atopic dermatitis (cAD), a clinical syndrome expanded from viewing cAD as a histamine-driven with intricate pathogenesis. Canine Atopic dermatitis Type I hypersensitivity to a very complex multifaceted results from the complex interaction between genetic syndrome. and environmental factors. It is becoming clear that Source of funding: Self-funded. cAD is not a single disease but rather a clinical Conflict of interest: None declared.

State-of-the-Art Address in 2013. In little more than seven years, recent papers Meticillin-resistant Staphylococcus have now compared NGS data for large numbers > pseudintermedius – this is your life ( 100) of MRSP isolates. Whilst the population structure of MRSP is highly clonal, many MRSP clones † D.J. TROTT*, J.M. NORRIS , J. KOVAC* and S. have acquired important genes such as those encoding ‡ ABRAHAM antimicrobial resistance through multiple, independent, *Australian Centre for Antimicrobial Resistance horizontal gene transfer events. Furthermore, strains Ecology, School of Animal and Veterinary Sciences, The within the epidemiological successful clones ST71 and University of Adelaide, South Australia, Australia; ST68 contain unique prophage-mediated resistance and † School of Veterinary Science, University of Sydney, virulence genes that may aid in fitness and clonal ‡ New South Wales, Australia; Antimicrobial Resistance spread. In all ST71 strains, prophages inserted into and Infectious Diseases Laboratory, School of genes affecting natural competency, prevent acquisition Veterinary Life Sciences, Murdoch University, Murdoch, of horizontal gene flow by genetic transformation and Western Australia, Australia may further drive clonal spread. These results, only In the year 2020, meticillin-resistant Staphylococcus made possible through breakthroughs in the analysis of pseudintermedius (MRSP) is officially 21 years of age. the pangenome, are shedding new light on the reasons Although first reported in Spain in 1995, it was not for MRSP’s “global success”. until 1999 that a MRSP strain, isolated from a dog in Source of funding: Research on Australian isolates of the United States, was confirmed to contain the mecA MRSP was supported by ARC Linkage Grant gene. At such an auspicious coming of age, it is timely LP130100736 with Zoetis and Luoda Pharma as the to look back on the key developments in our under- main Linkage Partners. standing of MRSP evolution as well as crystal-ball-gaze Conflicts of interest: DJT has received research funding into the future now MRSP has become endemic in from Luoda Pharma, Neoculi, Zoetis, Bayer, Boehringer most countries. This review will cover the key aspects Ingelheim, Virbac and Elanco. of MRSP’s “birth” and “adolescence” (i.e. molecular genetics, clonal diversity and epidemiological spread) and summarise some of the possible reasons for its Supporting Review “global success”. Key “newsworthy” events, cautionary Using a One Health approach to chart a path tales and successful treatment of problem cases out of the Pandemic Era throughout MRSP’s “life” will be explored, including J. MAZET development of resistance to critically important and Department of Epidemiology and Disease Ecology, topically applied antimicrobial agents. Given that Syd- School of Veterinary Medicine, University of California- ney NSW was to be the location for the World Veteri- Davis, Davis, CA, USA nary Dermatology Conference, a uniquely Australian view on MRSP also will be included. It will conclude Threats posed by pandemics and epidemics are now with a summary of what we have learned thus far and clear to every household around the world. A single what technological developments such as Next-Genera- lethal microbe can emerge suddenly and spread rapidly tion Sequencing (NGS) and rapid susceptibility testing to every community without regard to national borders are available to help us devise the treatment and con- or to social and economic standing. The SARS, Ebola trol plans of the future. and Zika outbreaks did little to prepare us for the The first bacterial genome (Haemophilus influenzae) was COVID-19 pandemic sweeping the world, which serves sequenced in 1995. In 2005, the estimated cost for gen- as a clarion call that we are vulnerable to emerging erating a bacterial genome sequence was approximately viral threats. Since the mid-20th Century, new and AU$50,000. Since then, the availability of rapid and deadly diseases have emerged at an alarming rate in accurate NGS platforms such as Illumina MiSeq and animals and people, and the threats from this vast pool an array of free, online bioinformatics tools has almost of unknown viruses are accelerating exponentially, dri- made whole genome sequencing cheap enough to ven by our expanding population and an increasingly request as a diagnostic test on a single isolate. Illumina interconnected world. After a decade of employing a MiSeq generates large numbers of short DNA reads One Health approach to viral detection, discovery and ideal for quickly and accurately interrogating the gen- characterization, we can reasonably estimate that there ome for multilocus sequence types, antimicrobial resis- are >500,000 zoonotic viruses, about which we know tance and virulence genes. Additionally, screening nothing or very little, that have the potential to spill multiple genomes for single nucleotide polymorphisms over from evolutionary host species to vulnerable ones and accessory genes provides unprecedented power for and cause disease. We know how to identify nearly all understanding clonality, epidemiological spread and the of the viral diversity in every species and can begin to influence of genetic recombination on MRSP popula- rank the risks of these viruses for interspecies transmis- tion structure. More expensive deep sequencing, how- sion. Understanding the disease threats and developing ever, is still required to close genomes and to mitigation strategies to prevent infection and spread of understand the genomic context of identified mobile viruses are key to preventing future devastating pan- genetic elements. The first reported fully annotated demics. The Global Virome Project is a strategic MRSP genome sequence, from a ST71 strain isolated response to the growing need to better predict, prevent from a canine bite wound in Denmark, was announced and respond to future viral pandemic threats, and to

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 Abstracts 9 protect us all from their worst consequences. By bring- will gain the benefits of shared global and regional ing health professionals and scientists from many disci- virus identification and disease mitigation strategies, plines together to share protocols and data under one while maintaining autonomy to respond to local needs. umbrella, a global consortium can be established to Source of funding: Self-funded. implement a joint coordinated effort, whereby countries Conflict of interest: None declared.

State-of-the-Art Address migration cues. Grhl3 null mouse embryos fail to heal Grainy head-like genes are key epidermal wounds and keratinocytes with reduced Grhl3 expres- regulators across 360 Myr of evolution sion display chaotic migratory patterns. In squamous epithelium from any tissue source, Grhl3 regulates the † Z. DENG*, S. M. JANE*, M. CANGKRAMA and balance between proliferation and differentiation, and M. R. CARPINELLI* its loss induces squamous cell carcinoma. In the skin, *Department of Medicine, Central Clinical School, this is mediated through increased levels of miR21, Monash University, Melbourne, Victoria, Australia; which reduces the expression of Grhl3 and its direct tar- † Institute of Molecular Health Sciences, ETH Zurich,€ get, PTEN leading to activation of the PI3K-AKT sig- Switzerland nalling pathway. Taken together, these data position There has been strong selective pressure to maintain a the GRHL family as master regulators of epidermal functional skin barrier since terrestrial animals evolved homeostasis across a vast gulf of evolutionary history. 360 Million years ago. In order to sustain terrestrial liv- Source of funding: Australian National Health and Medi- ing habits, these animals acquired a unique integumen- cal Research Council. tary system with a keratinized, stratified, squamous Conflict of interest: None declared. epithelium surface barrier. The barrier performs inside- out roles safeguarding against dehydration, and out- side-in roles preventing entry of microbes and toxins. Supporting Review The skin barrier centres on the stratum corneum layer An approach to cornification and hereditary of the epidermis, and consists of cornified envelopes cornification disorders in dogs cemented by the intercorneocyte lipid matrix in a E. MAULDIN bricks-and-mortar-like scenario. Multiple components Department of Pathobiology, School of Veterinary of the barrier undergo cross-linking by transglutami- Medicine University of Pennsylvania, Philadelphia, PA, nase (TGM) enzymes, while keratins provide additional USA mechanical strength. Cellular tight junctions also are crucial for barrier integrity. Our understanding of the The orderly formation of the stratum corneum (SC) is molecular basis of barrier formation, maintenance and vital to the health of the skin and body. The SC pro- repair is still evolving. Previous studies have identified vides the ultimate barrier between the vertebrate body highly conserved grainy head-like (GRHL) transcrip- and the ambient environment. It serves to restrict water tion factors that regulate the formation and mainte- flow in and out of the body to maintain hydration and nance of the integument in diverse species across prevent ingress of pathogens and toxins. The barrier is 700 Myr of evolution. Grhl3 is essential for formation comprised of terminally differentiated keratinocytes of the skin barrier during embryonic development and (corneocytes) that are sandwiched between lipid bilay- null mice die soon after birth from excessive transepi- ers. Primary disorders of cornification arise from Men- dermal water loss. The Grhl3 null epidermis is abnor- delian defects in the structural proteins, lipids and mally thick and shows failed terminal differentiation of enzymes that normally orchestrate formation of the keratinocytes accompanied by excess proliferation. skin barrier. Tremendous innovations in molecular Tgm1, encoding a transglutaminase, is a direct target of techniques have led to the identification of many disor- Grhl3 in epidermis, and loss of the cross-linking func- ders in both pure-bred and mixed breed dogs over the tion of this enzyme underpins the barrier defect. Sur- past decade. These advances have far outpaced the gen- prisingly, postnatal inactivation of the Grhl3 gene in eration of therapeutics. Disorders of cornification are conditional knock-out mice showed that it was dispens- managed rather than cured. Awareness of the lesion able for maintenance of the skin barrier, once formed. pattern, character and age of onset will help avoid pit- This was due to compensatory upregulation of Grhl1, falls in diagnosis. Disorders of cornification can be gen- which activates another transglutaminase, Tgm5, thus eralized (e.g. autosomal recessive ichthyosis) or mediating adequate cross-linking. Concomitant loss of localized to paw pads (e.g. palmar plantar kerato- Grhl1 and Grhl3 from postnatal mouse skin led to com- derma) or nasal planum (e.g. hereditary nasal paraker- plete regression of the formed skin barrier and was not atosis). Some disorders mimic those in humans while compatible with life. These findings identify the GRHL others are unique to the dog. This review will focus on factors as potential therapeutic targets in human dis- the epidermal structure and function in the process of eases in which barrier formation and function is com- cornification and specific defects that have been identi- promised. In addition to its barrier function, Grhl3 also fied in canine species. plays key roles in wound repair and as an epidermal Source of funding: Self-funded. tumour suppressor. In its former role, Grhl3 activates Conflict of interest: None declared. the planar cell polarity signalling pathway to mediate epidermal wound-healing by providing directional

State-of-the-Art Address of CH. The same VLP approach was used for induction Advances in diagnosis and therapy of equine of anti-IL31 antibodies in CH-affected horses and Culicoides hypersensitivity resulted in a decrease of CH lesions scores. These recent advances should lead to the development of bet- E. MARTI ter in vitro diagnostic tests and new treatment options Department of Clinical Research and Veterinary Public for CH, and are worthwhile studying for other equine Health, Vetsuisse Faculty, University of Bern, Bern, allergic diseases. Switzerland Source of funding: EM is funded by the Swiss National Culicoides hypersensitivity (CH) is an allergic, Science Foundation, the Morris Animal Foundation and immunoglobulin (Ig)E-mediated, Th2-type dermatitis of Stiftung Forschung fur€ das Pferd. horses caused by bites of insects of the genus Culi- Conflict of interest: None declared. coides. CH occurs worldwide except in Iceland. The prevalence of CH is particularly high in horses born in Iceland and exported to Culicoides-rich environments Supporting Review as adults. The performance of in vitro diagnostic tests Bacteriophage therapy and the efficacy of allergen-specific immunotherapy R. A. SQUIRES (ASIT) is questionable. This probably results from the Discipline of Veterinary Science, James Cook University, use of crude whole-body extracts from laboratory-bred Townsville, Queensland, Australia Culicoides species. For these reasons, salivary gland allergens from Culicoides have been identified and Bacteriophages (or just phages) are naturally occurring almost 30 produced as pure recombinant (r-) proteins. viruses that infect and kill bacteria. They are remark- These r-Culicoides allergens bind IgE from sera of CH- ably numerous, diverse and easily found, for example affected horses with varying frequencies. Protein in sewage and dog faeces. Each phage typically has a microarrays represent efficient tools for determination narrow host range, infecting only certain strains of a of the IgE sensitization pattern to allergens also in particular bacterial species. Yet a large majority of bac- horses. Testing of sera from horses living in various terial species studied to date play host to bacterio- European countries has resulted in the identification of phages, explaining the remarkable diversity. Phages the most relevant r-Culicoides allergens for CH. A com- were discovered just over 100 years ago and were first bination of seven major r-Culicoides allergens allows a used to treat bacterial infections in domestic animals specific (95%) and sensitive (90%) serological diagnosis and people in the 1920s. Phages grown in simple broth of CH. cultures of host bacteria were used successfully to treat A preventive ASIT is under development with the aim salmonellosis in chickens, bacillary dysentery in chil- of decreasing the incidence of CH in horses exported dren, and cutaneous furuncles and carbuncles in adult from Iceland. An immunization protocol was estab- humans. Phages have been safely administered orally, lished in Culicoides-unexposed horses living in Iceland. topically, intravenously, directly into infected lesions Immunization into the submandibular lymph node with and via inhaled aerosol. In many parts of the world, a combination of r-Culicoides allergens, using Alum interest in bacteriophage therapy waned after penicillin and MPLA as adjuvants, is able to induce an IgG became available. Interest continued in France and response without induction of IgE. The induced anti- parts of Eastern Europe. In Georgia, phages continue bodies are able to block binding of IgE to these same to be used to treat human bacterial infections, for allergens, an important feature of successful ASIT. Fur- example diabetic foot ulcers. During the 1980s, veteri- thermore, immunization with Alum/MPLA led to a narian H. Williams Smith and colleagues investigated higher production of interferon (IFN)-( and interleukin phage therapy in the UK. They sought to prevent and (IL)-10 than with Alum only, indicating that this com- treat severe Escherichia coli diarrhoea in calves, piglets bination of adjuvants is best suited for ASIT in horses. and lambs. In 1987, they showed that severe, experi- The efficacy of a preventive ASIT against CH will be mental E. coli diarrhoea in calves could be cured by a evaluated in a cohort of Icelandic horses exported from single dose of 100,000 phage particles and could be pre- Iceland to Switzerland. Therapeutic monoclonal anti- vented by doses as low as 100 particles. They obtained bodies (mAb) targeting key cytokines or their receptor their phages from ordinary sewage. The worsening are gaining in importance for treatment of human and antimicrobial resistance crisis has recently led to resur- also canine allergy. This approach, however, may not gent interest in bacteriophage therapy. Importantly, be available for equine patients due to the high amount phages can kill their host bacteria regardless of whether of mAbs required. A new approach was thus devel- or not the host is antimicrobial drug-resistant. This is oped: By contrast to passive immunization with mAb, because drug resistance usually has nothing to do with horses were actively immunized against selected cytoki- phage resistance. There recently has been a marked nes, linked to virus-like particles to enable antibody increase in the number and quality of research publica- production against a self-antigen. The first cytokine tar- tions dealing with phage therapy, mostly for use in geted using this approach was IL-5, important for the human medicine but also in veterinary science. A few differentiation, migration, activation and survival of promising publications have described successful, topi- eosinophils, which play an important role in the patho- cal phage therapy of Pseudomonas aeruginosa otitis genesis of CH. This therapeutic vaccine induced anti- externa (OE) in dogs. Consequently, a veterinary phage IL-5 antibodies, was safe, reduced eosinophil counts product for treatment of Pseudomonas OE in dogs and significantly improved clinical signs of CH in com- (containing six different bacteriophage strains) was parison to the placebo group. IL-31 is involved in the licensed and became commercially available in Europe. induction of pruritus and strong pruritus is a hallmark There have been some proof-of-concept studies. A New

Zealand-based research group showed that bacterio- for nearly 100 years, surprisingly little has been pub- phages readily found in sewage could kill a large pro- lished about their use in companion animals. What has portion of different uropathogenic canine and feline been published looks promising. Bacterial skin diseases E. coli strains. A very recent publication described the of dogs (including otitis externa) have been studied the isolation and characterisation of phages with preferen- most, although only to a small extent at this stage. tial activity against meticillin-resistant Staphylococcus Phage therapy represents a promising and potentially pseudintermedius strains, mainly from Europe and fruitful area of study for companion animal researchers North America. The anti-staphylococcal phages were in the future. isolated from canine faeces. Despite the fact that bacte- Source of funding: Self-funded. riophages have been used to treat bacterial infections Conflict of interest: None declared.

State-of-the-Art Address Improved understanding of the general clinical and his- Pathogenesis and treatment of alopecia in tological presentations in the early, mid- and late stages humans of these disorders will facilitate their understanding. Management strategies for these conditions are limited R. SINCLAIR yet a number of biologics therapy show promise in the Sinclair Dermatology, Melbourne, Victoria, Australia management of cicatricial alopecia. Hair is a defining characteristic of mammals. The hair Source of funding: Self-funded. follicle is a complete mini-organ that lends itself as a Conflict of interest: None declared. model for investigation of a variety of complex biological phenomena including stem cell biology, organ regeneration and cloning. The hair follicle has a com- Supporting Review plex immunology with constitutive expression of human Molecular mechanisms of noninflammatory leucocyte antigen G (HLA-G), yet low or absent alopecia in dogs expression of class 1-HLA. This is the basis of hair fol- † ‡ M. M. WELLE*, S. RUFENACHT€ , T. LEEB and licle relative immune privilege. Loss of this immune § D. J. WIENER privilege is thought to underlie alopecia areata (AA). *Institute for Animal Pathology, Vetsuisse Faculty, AA severity varies from a single small patch to com- University of Bern, Dermfocus, plete loss of scalp hair, body hair, eyelashes and eye- † Bern, Switzerland; DermaVet, Tierklinik Aarau West, brows. Approximately 40% of all affected individuals ‡ Oberentfelden, Switzerland; Institute of Genetics, only ever get one patch and will achieve a spontaneous, Vetsuisse Faculty, University of Bern, Bern, Switzerland; complete, durable remission within six months. AA that § Department of Veterinary Pathobiology, College of persists beyond 12 months is defined as chronic; with- Veterinary Medicine & Biomedical Science, Texas A&M out treatment, 55% of individuals with chronic AA will University, TX, USA have persistent multifocal relapsing and remitting disease and 45% will ultimately develop alopecia totalis or Lifelong recurrent hair growth is needed to renew the universalis. The unpredictable course contributes to the protective covering of the body and to enable adaption psychological distress attributable to AA. Numerous of coat quality and colour to environmental changes. topical, intralesional and systemic agents are currently An intact hair coat is provided by hair follicle morpho- used to treat AA; however, Janus kinase-signal trans- genesis during embryonic development and subse- ducer and activator of transcription (JAK/STAT) inhi- quently maintained postnatally by the hair cycle, bitors are emerging as a safe, effective treatment for during which hair follicles undergo recurrent phases of moderate to severe chronic AA. Other common causes growth (anagen), regression (catagen) and relative proof alopecia in humans are androgenetic alopecia liferative quiescence (telogen). The hair shaft is shed in (AGA) and cicatricial alopecia. AGA is a disorder per- a phase called exogen. At the end of telogen, new ana- petuated by androgens that occurs only in genetically gen is induced by the activation of stem cells, located in predisposed individuals. The pivotal role of androgens the secondary hair germ and in the bulge. If anagen and more specifically 5a reductase, the enzyme that induction does not occur, the telogen hair follicle will converts testosterone into dihydrotestosterone (DHT), eventually lose the club hair and a hair follicle in the has been known since the mid-1970s. However, the quiescent kenogen stage remains until appropriate sig- genetic and epigenetic bases of AGA are still not fully nals initiate a new anagen phase. Hair follicle morpho- elucidated. In addition the paradoxical site specificity genesis during embryogenesis and the maintenance of of the androgen effect on hair growth remains an the hair cycle during postnatal life are tightly regulated enigma. The search for a cure for baldness has led to and highly conserved processes, involving numerous significant recent advances in fundamental hair science. signals derived from epithelial, neuroendocrine and These include the description of scalp follicular units, mesenchymal cells. These signals are assembled in the the hierarchy of androgen sensitivity within follicular hair follicle stem cells and the interplay of all factors units and the central role of the arrector pili muscle in belonging to several signalling pathways finally result maintenance of follicular unit integrity. Current treat- in stem cell quiescence or activation. Once activated, ments for AGA include finasteride, dutasteride, minoxi- stem cells give rise to rapidly proliferating transient dil and prostaglandin analogues. Emerging treatments amplifying cells, which terminally differentiate into the include low-level laser, platelet-rich plasma injections different hair follicle layers and the layers of the hair and microneedling. The cicatricial alopecias include a shaft. In dogs noninflammatory alopecia can be subdi- poorly understood group of hair disorders linked by a vided into four categories: (i) Inherited alopecia with common final pathway of permanent hair follicle loss an impaired hair follicle morphogenesis affecting either and replacement by fibrous tissue in involved areas. hair follicle induction, organogenesis or cytodifferentia- There are several well-recognized secondary causes of tion during embryonic development; (ii) Alopecia with hair loss including trauma, infiltration (e.g. metastatic impaired hair follicle or hair shaft regeneration due to tissue) and infection. Primary cicatricial alopecias result a genetic predisposition and postnatal clinical manifes- from pathology centred on the hair follicle itself, rather tation; (iii) Acquired alopecia associated with an than the scalp, and include lichen planopilaris, discoid impaired hair cycle and eventual subsequent follicular lupus erythematosus, frontal fibrosing alopecia, dissect- atrophy (hair cycle arrest); and (iv) Acquired alopecia ing cellulitis and folliculitis decalvans. Progress in our associated with factors such as trauma, ischaemia and understanding and treatment of these disorders has poor nutrition. Some of the inherited congenital alope- been slow. Recently sunscreen use has been implicated cias represent monogenic traits that are caused, for in the development of frontal fibrosing alopecia. example, by variations in the FOXI3 or SGK3 genes. In

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 14 Abstracts genetically predisposed noninflammatory alopecias, the and the supporting review the following topics will be mode of inheritance appears to be more complex. Tran- addressed: (i) molecular regulation of the hair cycle; (ii) scriptome analyses in these disorders reveals different genetic variants and associated molecular mechanisms deregulated genes encoding for follicular stem cells, underlying inherited alopecias; and (iii) deregulation of proteins involved in Wingless-related integration site, genes and the associated consequences in alopecia Sonic hedgehog and bone morphogenic processing sig- developing postnatally. nalling, as well as proteins important for Vitamin D Source of funding: Self-funded. metabolism and steroidogenesis. In the presentation Conflict of interest: None declared.

SOS-1 NB-UVB MED attenuates Th1 or Tc cell-mediated skin inflammation, minimising epidermal damage in a Narrow-band ultraviolet B therapy attenuates canine model of hapten-induced contact dermatitis. Our immunological responses in hapten-induced findings implicate that NB-UVB phototherapy can be a contact dermatitis in beagles safe adjunct therapy for canine skin diseases associated S. ONISHI-SAKAMOTO*, K. MAKISHI*, K. with Th1 or Tc cells, such as the chronic phase of TAKAMI†, R. ASAHINA‡, S. MAEDA§,M. canine AD or canine epitheliotropic lymphoma. NAGATA¶, P. F. MOORE**, K. IDE* and K. Source of funding: Self-funded. NISHIFUJI* Conflict of interest: KT is an employee of Ushio Inc. *Laboratory of Veterinary Internal Medicine, Tokyo University of Agriculture and Technology, Tokyo, Japan; †Ushio Inc., Tokyo, Japan; ‡Graduate School of Medicine, Kyoto University, Kyoto, Japan; §Faculty of SOS-2 Applied Biological Sciences, Gifu University, Gifu, ¶ Cytokine expression in feline allergic Japan; Synergy Animal Hospital, Saitama, Japan; dermatitis and feline asthma **School of Veterinary Medicine, University of ¶ ¶ California Davis, Davis, CA, USA C. E. OLDER*, A. B. DIESEL , J. C. HESELTINE , A. FRIEDECK¶, C. HEDKE§, S. PARDIKE§,K. In human medicine, narrow-band ultraviolet B (NB- † ‡ § BREITREITER , M. A. ROSSI , J. MESSAMORE , UVB) phototherapy is known as a safe and effective § § G. F. BAMMERT , A. J. GONZALES and A. therapy in various inflammatory skin diseases such as RODRIGUES HOFFMANN* atopic dermatitis (AD) and early phase epidermotropic *Department of Veterinary Pathobiology, College of T-cell lymphoma. Phototherapy causes cell death of Veterinary Medicine & Biomedical Sciences, Texas immune cells in the skin while minimising tissue dam- A&M University, College Station, TX, USA; age. To obtain efficacious NB-UVB therapy, minimal ¶ Department of Small Animal Clinical Sciences, College erythema dose (MED), which is a minimal dose of of Veterinary Medicine & Biomedical Sciences, Texas UVB irradiation that causes gross erythema on human A&M University, College Station, TX, USA; skin, is applied to the lesional skin. However, the effect § Veterinary Medicine Research and Development, Global of NB-UVB on the cutaneous immune cells in dogs has Therapeutics Research, Zoetis Inc, Kalamazoo, MI, not yet been analysed. The purpose of this study was to † USA; South Austin Cat Hospital, Austin, TX, USA; investigate the effect of NB-UVB therapy on hapten- ‡ Coastal Veterinary Dermatology & Ear Clinic, induced contact dermatitis in dogs. To determine MED Houston, TX, USA of NB-UVB in the individual dog, seven healthy beagles were treated with NB-UVB doses of 300, 560, 750, The pathogenesis of feline allergic dermatitis is unclear, 920 and 1,000 mJ/cm2. The average of the NB-UVB with several differences from allergic dermatitis in dogs MEDs in six of seven dogs was 785 Æ 88 mJ/cm2 and humans. Few treatment options exist, and those (range: 560–1,000 mJ/cm2), although one dog did not available may not be effective and may have adverse exhibit gross erythema with 1,000 mJ/cm2 of NB-UVB. effects. To further understand the pathogenesis of feline To determine whether NB-UVB MED affects the allergies, we performed a survey of various cytokines immunological responses in inflammatory skin of dogs, within cats affected with allergic dermatitis and nonaf- 2,4-dinitrochlorobenzene (DNCB) was applied topically fected cats. We hypothesized that cats with allergic der- to the skin of six healthy beagles. DNCB is known to matitis would have higher cutaneous expression of cause type 1 helper T cell (Th1)- and/or cytotoxic T cell interleukin (IL)-31 and IL-31 receptor alpha (IL-31RA) (Tc)-induced skin inflammation. Treated skin then was than nonaffected controls, given their role in canine irradiated by NB-UVB MEDs. The skin samples were and human atopic dermatitis (AD). To measure cyto- subjected to immunohistochemical analysis as well as kine levels, formalin-fixed, paraffin-embedded skin quantitative reverse transcription-PCR for cytokine biopsies from cats with and cats without allergic der- transcription analysis. As a result, numbers of dermal matitis were obtained from two service archives. Using CD4+ and CD8+ positive cells in irradiated skin quantitative PCR, mRNA expression was evaluated for (185.0 Æ 58.3 cells/mm2 and 508.7 Æ 217.2 cells/mm2, IL-31 and IL-31RA. Using RNAScope, mRNA expres- respectively) were significantly lower than those in non- sion of IL-5, IL-31, IL-31RA, IL-33 and Oncostatin M irradiated skin (287.2 Æ 84.5 cells/mm2 and receptor (OSMR)-beta, part of the heterodimeric recep- 844.7 Æ 286.9 cells/mm2; Wilcoxon signed rank test, tor for IL-31, also was evaluated in tissues. Addition- P < 0.05). TdT-mediated dUTP nick-end labelling ally, IL-31 protein levels were evaluated via revealed that dermal apoptotic cells were significantly immunohistochemistry (IHC). IL-31 and IL-31RA were increased in irradiated skin (47.6 Æ 12.8 cells/mm2) low or not detected with qPCR in most samples. RNA- compared to those in nonirradiated skin scope corroborated qPCR results indicating extremely (26.1 Æ 6.8 cells/mm2; Wilcoxon signed rank test, low levels of mRNA expression for IL-31 and IL31RA. P < 0.05). Frequency of apoptotic keratinocytes in the IL-5 and IL-33 also were expressed in low quantities. irradiated skin was <0.5 cells/mm2 of basement mem- By contrast, OSMR-beta was highly expressed and branes. qRT-PCR revealed that transcription of genes increased in the skin of allergic cats versus healthy cats encoding interferon c- and Tumour necrosis factor-a, (P < 0.05). IHC revealed that IL-31 protein was which are Th1 cytokines, were significantly lower in expressed likewise in allergic and healthy cats. Both IL- irradiated skin than in nonirradiated skin (Wilcoxon 31 protein and mRNA were predominantly expressed signed rank test, P < 0.05). These findings indicate that by follicular epithelial cells and keratinocytes. Weaker

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 16 Abstracts expression also could be observed in muscle, sebaceous characterized dog-specific antibodies. The objective of and apocrine glands, and occasionally in infiltrating this study was to comparatively analyse, at both struc- inflammatory cells. We also investigated circulating tural and molecular levels, canine healthy and atopic levels of cytokines. In addition to healthy cats and skin. Six healthy and six spontaneously atopic labora- those affected by allergic dermatitis, patients with feline tory beagles housed in identical environmental condi- asthma – an incompletely understood allergic condition tions (CanCog Technologies; Ontario, Canada) were thought to be due to a Th2 immune response – were included in the study after validation of the experimen- sampled. We hypothesized that cats with either cuta- tal protocol by CanCog ethics committee (reference neous or respiratory allergic disease would demonstrate VRI185-18245-CO), and in accordance with the princi- higher levels of Th2 cytokines. For systemic measure- ples of the Animal for Research Act of Ontario and the ment of cytokine levels, serum was collected from a dif- guidelines of Canadian Council on Animal Care. Dogs ferent cohort of cats including 17 healthy cats, 18 cats were clinically characterized [general examination, with allergic dermatitis and 18 cats with asthma. Sam- Canine Atopic Dermatitis Extent and Severity Index, ples were subjected to a Luminex panel of 19 cytokines 4th iteration (CADESI-04) evaluation and transepider- covering both Th1- and Th2-associated molecules and mal water loss (TEWL) measurement] and skin biopsies an IL-31 ELISA. None of the cytokines were different were performed in healthy, lesional and non-lesional between healthy cats and those affected by allergic der- skin. Samples were analysed using haematoxylin & matitis and/or asthma. Interestingly, IL-31 was detected eosin staining and transmission electron microscopy in a greater proportion of samples from cats with aller- (TEM). The expressions of FLG, corneodesmosin gic dermatitis (10/18) than from healthy (4/17) and (CDSN), claudin-1 and involucrin were studied, using asthmatic cats (5/18). previously validated antibodies, by indirect immunoflu- These results suggest that feline patients suffer from orescence and Western blotting, as well as by quantita- allergic diseases that may exhibit different pathomecha- tive reverse transcription PCR. TEWL was significantly nisms from allergic diseases affecting other species, higher in atopic dogs when compared to controls, sug- which typically exhibit a Th2-skewed immune system. gesting skin barrier impairment. Blinded evaluation of Although IL-31 is shown to be relevant in human and H&E stained skin sections did not allow discrimination canine AD, the results of this study did not reveal dra- between the healthy and non-lesional atopic skin. TEM matic differences in IL-31 levels in tissue or serum, examination showed abnormal morphology of the stra- instead showing an upregulation of the IL-31 co-recep- tum corneum (SC) in the non-lesional atopic skin, tor OSMR-beta. These findings implicate aspects of the including thicker corneocytes (P = 0.02) with very irreg- IL-31 pathway that may be dysregulated and suggest ular morphology and a sometimes vesicular aspect. The that the co-receptor OSMR-beta has an important role SC compactum was either absent or of increased thick- in the pathogenesis of feline allergic skin disease. ness, when compared to the healthy skin. These Source of funding: Zoetis Inc. changes were even more pronounced in lesional skin. Conflicts of interest: CEO, ABD, JCH and ARH No significant differences were observed between received partial salaries from Zoetis Inc. CH, SP, JM, groups in the number nor the size of keratohyalin gran- GFB and AJG are employees of Zoetis Inc. ules (containing FLG and loricrin), nor in the number of corneodesmosomes (containing CDSN). Blinded qualitative evaluation of immunofluorescence reactivity by two investigators failed to show any significant dif- SOS-3 ferences between groups for any of the proteins studied. Structural and molecular comparative analysis This result was confirmed by an objective evaluation of grey levels using IMAGEJ software. Intensity of Western of normal and atopic canine skin blotting bands was normalized to actin and statistical D. COMBARROS*,†, D. GOUDOUNECHE‡,M.C. comparison between groups further confirmed the CADIERGUES*,† and M. SIMON§ absence of differences in protein expression (FLG, *Small Animal Clinic, Universite de Toulouse, Toulouse, CDSN, claudin-1 and involucrin). Expression also was France; †UDEAR, Universite de Toulouse, Toulouse, similar at the mRNA level. As in humans, epidermal France; ‡Centre de Microscopie Electronique Appliquee differentiation seemed to be impaired in canine AD, as a la Biologie, FacultedeM edecine Rangueil, Universite shown by both abnormalities of the SC structure and de Toulouse, Toulouse, France; §UDEAR U1056, high water permeability. However, the expression of Universite de Toulouse, Toulouse, France some major epidermal proteins was not found to be altered. Further studies including a larger number of The pathogenesis of atopic dermatitis (AD) is complex animals and use of additional validated antibodies and not fully understood. Similar to people, dogs directed against epidermal proteins are necessary to develop spontaneous AD, suggesting this species could determine the cause of these abnormalities in the SC. be a useful model of study. Skin barrier defects, such Source of funding: The project was partially financed by as filaggrin (FLG) gene loss-of-function mutations and the BQR 2019 research grant of the National Veterinary FLG decreased expression, are well-known in humans School of Toulouse (ENVT). and are considered to play a key role in the pathogene- Conflict of interest: None declared. sis of AD. However, little information exists concerning the expression of skin barrier-related proteins in dogs owing to the low number of studies and of well-

SOS-4 of atopic dogs despite subtle conjunctival clinical signs. As IL-8 is commonly found in many inflammatory con- Survey of cytokines on the ocular surface of ditions, further studies are needed to determine its atopic dogs by multiplex analysis using two specificity in atopic conjunctivitis. Although higher con- sampling methods: a pilot study centrations of markers were obtained with method A, C. PRESSANTI*,†, E. RAVAILHE*, J. BRUN*, N. method B gave similar results. The Eyeprim device is AMALRIC‡, L. A. LECRU*, J. KONDRAJTEVA*, easy to use and allows specific sampling from the con- F. MOOG*, D. COMBARROS*,†, J. Y. DOUET*,§ junctiva. Canine tears have been shown to contain and M. C. CADIERGUES*,† cytokines which are not necessarily linked to the *Small Animal Clinic, Universite de Toulouse, Toulouse, inflammatory status of the conjunctiva. The Eyeprim France; †UDEAR, Universite de Toulouse, Toulouse, device could be more discriminant than swabbing to France; ‡Synelvia, Labege, France; §IHAP, Universitede evaluate conjunctival markers and would limit tear col- Toulouse, Toulouse, France lection and be more representative of ocular status. Source of funding: Self-funded. Conjunctivitis in atopic dogs has been described and is Conflict of interest: None declared. likely underdiagnosed in practice. Consistent conjunctival infiltration by eosinophils in atopic dogs, as compared with healthy controls, was reported in a previous study of conjunctival biopsies as a possible hallmark of SOS-5 an allergic reaction. Elevated levels of inflammatory cytokines have been reported in tears from ocular aller- Interleukin-31 and IL-31 receptor expression gic disease states in humans. The purpose of this in acute experimental canine atopic dermatitis prospective study (Ethics Committee approval no. skin lesions SSA_2019_005) was to assay various cytokines in tears and conjunctivae from atopic and normal dogs, and to C. TAMAMOTO-MOCHIZUKI and T. OLIVRY compare canine atopic dermatitis-associated conjunc- Department of Clinical Sciences, College of Veterinary tivitis with controls. Ocular surfaces from 10 atopic Medicine, NC State University, Raleigh, NC, USA and 10 normal dogs were sampled bilaterally using two Interleukin (IL)-31 is one of the earliest and most different methods. Tear samples were obtained with a important pruritogenic cytokines in canine atopic der- sterile swab previously moistened with saline solution matitis (AD). To optimize the IL-31 signalling blockade (method A). Conjunctival impressions were obtained therapy in AD, an understanding of the chronological with a validated device (Eyeprim, Opia Technologies; changes in IL-31 and its receptor (IL-31RA) in canine Paris, France) (method B). For each sample, the con- skin lesions is needed. This study aimed to, after an centrations of a panel of cytokines, comprising inter- allergen challenge, (i) assess the chronology of the leukin (IL)-2, IL-6, IL-7, IL-8, IL-10, IL-15, IL-18, expression of IL-31 during experimental acute AD skin tumor necrosis factor (TNF)-alpha, interferon (IFN)- lesions and compare it with the serum IL-31 level, and gamma, granulocyte-macrophage colony-stimulating (ii) determine the identity of cells producing IL-31 and factor (GM-CSF), IFN-gamma–induced protein 10 (IP- expressing the IL-31RA. Skin and blood samples were 10), monocyte chemoattractant protein 1 (MCP-1) and obtained from four Maltese-beagle atopic dogs at 0 keratinocyte-derived chemokine (KC)-like, were mea- (baseline), 24, 48 and 96 h after epicutaneous house sured in duplicate by multiplex analyses (#CCYTMG- dust mite (HDM) application. Immunofluorescence was 90K-PX13, Merck Millipore; Darmstadt, Germany). performed with a Zoetis-developed mouse anti-canine- Canine Atopic Dermatitis Extent and Severity Index, IL-31 monoclonal antibody and goat anti-human-IL- 4th iteration (CADESI-04), pruritus (PS) and conjuncti- 31RA polyclonal antibodies. Each section also was val (CS) scores were determined. Cytokine levels were double-stained for IL-31/CD3, IL-31/CD4 and IL- compared by Wilcoxon–Mann–Whitney U-test; correla- 31RA/b3-tubulin (a neuronal marker). The IL-31-posi- tions with CS, PS and CADESI-04 were determined by tive round/dendritic cells were subjectively scored for Spearman rank correlation test. Median age in normal their distribution (0, none; 1, one focal area; 2, multifo- dogs was 5.3 years (range 1–11) and in atopic dogs 4.4 cal areas; 3, diffuse) and count (0, none; 1, low; 2, med- years (range 1–10). In atopic dogs, the median CS was ium; 3, high) to obtain an expression score (distribution 1.5/12 (range 0–7), the median CADESI-04 was 43.5 score x cell count score). The serum IL-31 level was (range 39–57) and the median PS was 10.2/16 (range measured using an ultrasensitive signal molecule array. 2.8–14.7). Among the measured biomarkers, only GM- A Spearman rank correlation test was used to investi- CSF, KC-like and IL-8 were above the limit of quan- gate the correlation between the IL-31 expression score tification in most samples. Absolute amounts of each and the serum IL-31 level. All of the procedures were biomarker were always higher in samples obtained with approved by our university’s Institutional Animal Care method A than with method B. GM-CSF amounts and Use Committee. On the one hand, the peak IL-31 were lower in atopic dogs (method A, P = 0.02; method expression scores for each dog occurred at 24 h and/or B, P=0.0005). KC levels were higher in atopic dogs, yet 48 h after HDM provocation; it started to decrease at the differences were not significant. IL-8 amounts were 96 h, except in Dog 3 whose score continued to higher in atopic dogs (method A: 378 and 1504 pg/mL increase throughout the study period. On the other, the in normal and atopic dogs, respectively, P = 0.0003; serum IL-31 levels for each dog increased throughout method B: 57 and 382 pg/mL in normal and atopic the study, except, again, for Dog 3 whose serum IL-31 dogs, respectively, P = 0.006). In atopic dogs, IL-8 con- level was unchanged. Overall, there was only a weak centrations obtained with method B were positively cor- negative correlation between the IL-31 expression score related with CS (P = 0.008) and PS (P = 0.036). on the skin and the serum IL-31 level (r = -0.25; Regardless of the method, these preliminary results sug- P = 0.35). The majority of IL-31-positive cells in the gest an overexpression of IL-8 in conjunctivae and tears dermis co-expressed CD3 (range 91–100%) and CD4

(range 63–100%), indicating that they were likely Th2 Dermatitis Extent and Severity Index, 4th iteration cells. Unexpectedly, sebaceous glands were strongly (CADESI-04) scores were evaluated every 30 days and stained with anti-IL-31 antibodies; the extinction of this analysed by ANOVA of repeated measurements positivity by pre-incubating the antibodies with recom- (P < 0.05). Sensitivity (SE), specificity (SP), positive binant canine IL-31 further supported the presence of predictive value (PPV), negative predictive value (NPV) IL-31 in sebaceous glands. The IL-31RA immunofluo- and accuracy (AC) and Kappa values between prick- rescence staining showed an epidermal intercellular pat- and-patch test were calculated. Twenty-one (55%) of tern at all time points, suggesting its expression by the 38 dogs had a positive reaction to at least one test keratinocytes. Some of the dermal b3-tubulin-positive and of these, 14 (66.7%) presented sensitivity to envi- linear structures were double-positive for IL-31RA, ronmental allergens. A significant average improvement indicating that it was constitutively expressed on dermal in pVAS and CADESI-04 scores was obtained by 17 nerves. Herein, we showed, in experimental acute (81%) of the 21 dogs after 30 (P<0.01) and 60 (P<0.01) canine AD skin lesions, the early and transient secre- days of dietary exclusion, and all 17 dogs presented sig- tion of IL-31 by helper T cells – likely Th2 – that does nificant increased scores after food allergen re-exposure. not parallel the serum IL-31 level. The detection of IL- There was no difference in pVAS and CADESI-04 31RA in keratinocytes and peripheral nerves in canine scores between D30 and D60. The combination of tests skin indicates the potential roles of IL-31 as a pro- showed values of SE, SP, VPP, VPN and AC of 80%, inflammatory factor involving keratinocytes and dermal 66.7%, 66.7%, 80% and 73%, respectively. There was nerves in the transmission of itch in atopic dogs. low kappa agreement (j = –0.17) between the two aller- Source of funding: Zoetis. gic tests. It is concluded that most dogs with chronic Conflict of interest: The PhD programme of CTM is pruritus associated with AD are polysensitized to food sponsored by Zoetis. and environmental allergens. The lack of agreement between the tests suggests the need to perform both. These tests can represent an important method for establishing dietary protocols and provocative testing, SOS-6 allowing an early diagnosis of AFR, with a 30 day elimination diet. Association of prick-and-patch test in the Source of funding: Coordenacßao~ de Aperfeicßoamento de evaluation of adverse food reaction in dogs Pessoal de Nıvel Superior (CAPES). with allergic dermatitis Conflict of interest: None declared. J. POSSEBOM, A. CRUZ, V. GMYTERCO and M. FARIAS Department of Veterinary Medicine, School of Life Sciences, Pontifıcia Universidade Catolica do Parana, SOS-7 Curitiba, PR, Brazil Intradermal immunotherapy with Adverse food reaction (AFR) in dogs is assessed Actinomycetales preparations as treatment for through an elimination diet for eight weeks, followed feline hypersensitivity dermatitis: a preliminary by food provocation, which often has poor compliance randomized, placebo-controlled, double-blinded by owners and variable results in diagnosis. Previous studies have shown that the patch test with food study extracts was adequate to assist formulations of elimina- D. SANTORO, L. ARCHER and L. FAGMAN tion diets in patients with suspected AFR. Association Department of Small Animal Clinical Sciences, College of prick-and-patch tests assist diagnosis of AFR in chil- of Veterinary Medicine, University of Florida, dren with atopic dermatitis (AD); however, little is Gainesville, FL, USA known about these tests in dogs. The aim of this study Feline hypersensitivity dermatitis (FHD) is an extre- was to evaluate the effectiveness of prick-and-patch test mely common skin disease affecting ≤33% cats seen by in guiding elimination diet protocols and in the food dermatology specialty practices. Causes of FHD include provocation in dogs with AD that have an AFR. The allergic dermatitis such as fleabite hypersensitivity, study was approved by the Ethics Committee of cutaneous adverse food reaction, urticaria, angioedema PUCPR (01252). Thirty-eight dogs with AD were and feline atopic syndrome. Although very common, selected on which to perform prick tests with food few therapeutic options are available for cats with extracts (bovine protein, chicken, pork, fish, egg, milk, FHD. Very recently, the use of a single intradermal soy, wheat, rice, potato and manioc) and with extracts injection of heat-killed Actinomycetales (Mycobacterium of dust mite and pollen allergens (Veterinary Allergen- vaccae, Gordonia bronchialis, Rhodococcus coprophilus ics; Rio de Janeiro, Brazil). The patch test was per- and Tsukamurella inchonensis) as bacterial formed using the same food sources, placed in ~ immunomodulators has shown potential beneficial Allergochambers containers (Neoflex; Sao Paulo, Bra- effects on canine allergies. To date, no studies have zil) and then kept for 48 h in contact with the skin, been published on the use of such therapy for FHD. and the presence of erythematous papules, spots or pla- Thus, the purpose of this double-blinded, randomized, ques were considered positive. Patients with positive placebo-controlled clinical trial was to evaluate the clin- reactions to at least one food extract in the prick test ical effects of heat-killed Actinomycetales and/or patch test were submitted to an elimination diet [G. bronchialis (GB) and R. coprophilus (RC) alone or and evaluated on day (D)0, D30 and D60. In patients in combination) in FHD. Privately owned cats with a that had improvement, a provocation test was per- diagnosis of FHD were included in this study. Cats formed with a food that had positive and a negative were randomly allocated in one of three treatment reaction, at D75 and D90, respectively. The pruritus groups (GB, RC and GB/RC combination) or placebo. Visual Analog Scale (pVAS) and Canine Atopic

Five intradermal injections [day (D)0, D20, D40, D90 exudative or proliferative according to clinical signs. and D180) were performed over a one year period. All previous treatments of EPD of the last six months Because of the duration of the study, rescue antipruritic were recorded and categorized as disinfectant, antibiotic mediations were allowed, if necessary, and frequency or no antibacterial. Skin swabs were obtained from the and doses recorded. At each visit (D0, D20, D40, D60, most severely affected pastern and one control pastern. D90, D180 and D360), the severity of the clinical signs DNA was extracted from the swabs and the bacterial (lesional score and pruritus), global owner assessment 16S rRNA gene region was amplified. Samples were score, use of rescue medications, clinical adverse effects, submitted for paired-end sequencing on an Illumina skin hydration and cutaneous pH were assessed. A MiSeq platform (Illumina Inc.; San Diego, CA, USA). Friedman’s test and a Kruskal-Wallis test followed by Alpha diversity (i.e. within-sample diversity) was signifi- Dunn’s multiple comparison test were performed to cantly reduced in affected pasterns in comparison to analyse the behaviour of each variable; P ≤ 0.05 was their respective controls as determined by a univariate considered statistically significant. A total of 17 cats Wilcoxon signed rank test (P = 0.001, r = 0.54). A mul- (GB: n = 6; RC: n = 3; GB/RC: n = 4; placebo: n = 4) tilevel mixed effects model showed that the Shannon were enrolled in this study. All cats except two (Sia- diversity index was significantly associated with the mese) were domestic short hair cats. The average (Æ form of EPD (P = 0.001) and not with the type of pre- SD) age at the time of enrolment was 5.7 Æ 3.5 years vious treatment (P = 0.16). Analyses of beta-diversity with no significant difference among groups (P > 0.6). (i.e. between-samples dissimilarity) showed a disorder- The median weight at enrolment was 4.8 kg with no ing of the microbiota in affected pasterns. A Procrustes significant difference among groups (P > 0.2) or over analysis revealed that proliferative EPD induced the time (P > 0.1). Compared to placebo, GB significantly largest alteration in microbiota, followed by exudative improved the clinical score from D90 (P < 0.05). In the EPD and mild EPD, with mean residual values of 0.2, GB group, an over-time clinical score reduction was 0.18 and 0.14, respectively. Much of this divergence seen from D60 (P ≤ 0.03). A complete remission of the was driven by staphylococci bacteria, with affected pas- clinical signs was seen in 30–67% of cats in all treat- terns having a significantly higher relative abundance ment groups at various time points. A significant reduc- than their controls (univariate Wilcoxon signed rank tion in the pruritus score was only achieved by RC on test, P = 0.011, r = 0.37). On average, the relative D360 (P = 0.04) when compared to D0. Significant dif- abundance of staphylococci increased by 7.8% in ferences in the other parameters were not seen. Adverse affected pasterns (SD: 18.1). However, the intraindivid- effects were not seen in any cat. In conclusion, the use ual increase varied greatly between horses, being partic- of intradermal injections of GB is an effective and well- ularly large in exudative EPD lesions and in pasterns tolerated treatment for FHD. Because of the low cost that had previously received antibacterial treatment. In and the lack of adverse effects, GB – alone or in com- conclusion, our study highlights the influence that EPD bination with other treatments – could offer a beneficial has on the pastern bacterial skin microbiota, confirm- treatment option for affected cats. A larger study is ing that bacteria are an important factor in the course needed to confirm these data and to evaluate the of this multifactorial disease. In addition, although immunological changes occurring in the treated cats. antibacterial treatment may not be the dominant factor Source of Funding: This study was funded by the Univer- for the overall reduction of bacterial diversity in EPD, sity of Florida Research Foundation. it might be a predisposing factor for the growth of Conflict of interest: None declared. staphylococci as opportunistic pathogens. Source of funding: Self-funded. Conflict of interest: None declared.

SOS-8 Investigation of the skin microbiota in equine SOS-9 pastern dermatitis Whole genome sequencing and de novo S. KAISER-THOM*, M. HILTY† and V. GERBER* *Swiss Institute of Equine Medicine (ISME), Vetsuisse assembly of Staphylococcus pseudintermedius: Faculty, University of Bern, Bern, Switzerland; †Institute a novel pangenome approach to unravel for Infectious Diseases, University of Bern, Bern, pathogenesis of canine pyoderma Switzerland L. FERRER*, R. FONTICOBA*, D. PEREZ †,J. Equine pastern dermatitis (EPD) is a multifactorial syn- VINES~ †, S. MADRONERO~ †, A. CUSCO†,L. drome that has variable degrees of severity. Despite the MIGURA‡ and O. FRANCINO† widespread use of antibiotics in horses with this condi- *Department de Medicina I Cirurgia Animals, tion, the role of bacteria has not been adequately inves- Universitat Autonoma de Barcelona, Spain; †Servei tigated. This study explored the bacterial skin Veterinari de Genetica Molecular, Universitat Autonoma microbiota of affected (EPD) and control pasterns, and de Barcelona, Spain; ‡Centre de Recerca en Sanitat investigated associations with the clinical presentation Animal, IRTA, Bellaterra, Barcelona, Spain and previous treatments. The study protocol was The majority of canine pyodermas are caused by Sta- approved by the veterinary ethical committees of all 26 phylococcus pseudintermedius, which is part of the nor- cantons in Switzerland. The study design was case con- mal skin microbiota of the dog. The mechanisms that trol. All legs of each horse were examined with a stan- lead a commensal bacterium to transform itself into a dardized procedure. One affected and one control- pathogen have so far remained elusive to research. suitable pastern in the same horse was a prerequisite However, understanding the whole pathogenic process for participation. Eighty horses fulfilled the inclusion is of outmost importance to develop new treatment and criterion. Affected pasterns were classified into mild, prevention strategies. Whole genome sequencing (WGS)

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 20 Abstracts is regarded currently as the most comprehensive way of SOS-10 obtaining relevant genomic information of micro- organisms. We hypothesized that comparative analysis The efficacy of olanexidine gluconate in canine of the complete genome of S. pseudintermedius strains superficial pyoderma: in vitro and in vivo isolated from healthy dogs and dogs with pyoderma analysis could help to understand the pathogenesis of pyoderma. The aims of this study were (i) to visualize and Y.H. HSIAO, K. SHIMOIKE and K. IYORI understand the pangenome of S. pseudintermedius (core Vet Derm Tokyo, Kanagawa, Japan genome and accessory genome) and (ii) to find differ- Superficial pyoderma caused by meticillin-resistant Sta- ences between strains isolated from healthy and affected phylococcus pseudintermedius (MRSP) is an emerging dogs. Sequencing was performed using Oxford Nano- problem and it often leads to limited options for sys- pore MinION, a new technology that delivers longer temic antimicrobial therapy. By contrast, topical reads (up to tens or hundreds of kilobases) in a hand- antiseptic treatments such as chlorhexidine shampoo or held device. The long reads obtained allow de novo solution have been reported to be effective treatment assembly of bacterial genomes, and its portability and options for MRSP skin infections in dogs. Recently, a real-time results are essential to ultimately develop a novel biguanide of olanexidine gluconate (Olanedine, clinical metagenomic approach to canine pyoderma. Otsuka Pharmaceutical Co., Ltd; Tokyo, Japan) was Five strains of S. pseudintermedius isolated from the launched as a broad spectrum bactericidal solution for abdominal skin of five healthy dogs and 27 strains iso- disinfecting human skin. However, olanexidine glu- lated from lesional skin of 27 dogs with pyoderma were conate has not been evaluated sufficiently in canine analysed. DNA was extracted using the DNA Micro- superficial pyoderma. The aim of the study was to prep kit (ZymoBIOMICS). DNA quality was assessed investigate the clinical utility of olanexidine gluconate with a Qubit fluorimeter. The libraries for Nanopore compared with topical chlorhexidine and determine its sequencing were prepared by transposase fragmentation minimum inhibitory concentration (MIC) in S. pseudin- using the Rapid Barcoding kit and sequenced in a termedius. In an in vivo study, 33 client-owned dogs MinION FLO-MIN106 v9.4.1 flow cell. The fast5 reads with superficial pyoderma caused by S. pseudinter- were basecalled with Guppy, assembled with Flye, and medius infection were recruited. Species identification corrected with racon and medaka. The pangenome was and meticillin resistance were detected using a multiplex analysed and visualized with Anvi’o 6.1. Nanopore PCR method for identifying the thermonuclease and sequencing was effective and successful and allowed the mecA genes. Dogs were allocated randomly to olanexi- assembly of the complete genome of S. pseudinter- dine or chlorhexidine treatment groups. In the olanexi- medius (2.69 Mb; range 2.55–2.89 Mb) in a single con- dine group (n = 15), the dogs received a gauze-dipping tig. The mean genome size of S. pseudintermedius treatment of 1.5% olanexidine gluconate solution on strains isolated from healthy dogs (2.55 Mb) was smal- the lesion sites once daily. In the chlorhexidine group ler than the mean size of strains isolated from lesions (n = 17), dogs received 3% chlorhexidine gluconate of dogs with pyoderma (2.71 Mb). The Anvi’o platform shampoo (Douxo, CEVA; Lenexa, KS, USA) once a allowed an accurate visualization of the core and acces- week according to the product instruction. All dogs sory genomes, and the detection of differences between were not allowed antibiotics or anti-inflammation medi- commensal and pathogenic strains. The genome of cations either systematically or topically. Clinical scores S. pseudintermedius is an example of an open pangen- of superficial pyoderma were evaluated day (D)0 and ome. The core genome represents 45% of the total and D14 following the guidelines of the Japanese Society of is present in all of the strains (housekeeping genes). Antimicrobials for Animals. All investigators were The accessory genome (55%) is more prominent in the blinded to treatment allocation. All patients were able pathogenic strains and comprises antimicrobial resis- to finish the trial, and MRSP was identified in seven tance genes, virulence factors and bacteriophages. The dogs in the olanexidine group and 10 dogs in the analysis of the pangenome of S. pseudintermedius is chlorhexidine group. At D14, clinical scores were signif- very promising as a tool to understand the pathogenesis icantly reduced compared to D0 in both groups of canine pyoderma and the transformation of a com- (P < 0.05). The improvement rate of clinical score at mensal into a pathogen. Recent studies have demon- D14 was 46% and 43% in the olanexidine and strated the aetiological diversity and complexity of chlorhexidine groups, respectively; there was no signifi- canine pyoderma. Traditional culture techniques and cant difference in the improvement of clinical scores determination of antimicrobial sensitivity have limita- between the groups (P > 0.05). Two dogs developed tions facing this complexity. The analysis of the pan- mild scaling after one week of olanexidine application genome provides more accurate and clinically relevant and they recovered spontaneously after the trial. We information. further performed an in vitro study to determine the Source of funding: Research Project RTI2018-101991-B- MIC both in MRSP and meticillin-susceptible S. pseud- I00 (“From whole genome sequencing to clinical metage- intermedius (MSSP) isolated from dogs with superficial nomics: investigations on the pathogenesis of Staphylo- pyoderma. Fifty-seven strains of S. pseudintermedius coccus pseudintermedius pyoderma in the dog”), Spanish (29 MRSP and 28 MSSP strains) were included in the Ministry of Science, Innovation and Universities. in vitro study. In the broth microdilution method, Conflict of interest: LF has received unrelated honoraria MICs of olanexidine gluconate were 1.46 lg/mL in for lecturing from Zoetis, Bayer, LETI and Affinity Pet- MRSP and 0.73 lg/mL in MSSP. Therefore, 1.5% care. olanexidine gluconate solution contains 1027.4- to 2054.8-fold higher concentration than the determined MIC. From these findings, 1.5% olanexidine gluconate solution could be an effective and tolerable topical therapy option for dogs with superficial pyoderma even in MRSP infections.

Source of funding: Self-funded. should be used according to the results of susceptibility Conflict of interest: None declared. tests. Following the restricted use of antimicrobials, an antibiogram was reconstructed using the susceptibility test data between January and December 2019. Fifty- three SP strains were isolated from dogs with pyoderma SOS-11 (n = 30) and OE (n = 22). The susceptibility rates of MPIPC (60%), CEX (66%) and MINO (56%) in 2019 Advantages of restricting antimicrobial use were significantly higher than those between 2017 and with antibiograms for reducing antimicrobial 2018 (P < 0.05). The other drugs showed higher suscep- resistance rate in dogs with Staphylococcus tibility rates than those observed in 2017, with the pseudintermedius infections exception of ST (24%). These findings indicated that † the restricted use of antimicrobials for over a year, K. IYORI*, T. SHISHIKURA , K. SHIMOIKE*, K. based on antibiograms, reduced the rate of antibiotic MINOSHIMA* and Y. TOYODA* resistance of SP strains isolated from dogs with pyo- *Vet Derm Tokyo, Kanagawa, Japan; †Daktari Animal derma and OE. Hospital Tokyo Medical Center, Tokyo, Japan Source of funding: Self-funded. The antibiotic resistance of Staphylococcus pseudinter- Conflict of interests: None declared. medius (SP) and meticillin-resistant SP (MRSP) is increasing dramatically in dogs worldwide. An antibiogram is a periodic summary of the test results of antimicrobial susceptibility of specific micro-organisms SOS-12 to batteries of antimicrobial drugs. Antibiograms are used to select appropriate empirical antimicrobial treat- Persistent activation of the mammalian target ments and monitor the trends in antimicrobial resis- of rapamycin signalling pathway in cutaneous tance. This study evaluated whether the restricted use squamous cell carcinomas in cats of antimicrobials with antibiograms can reduce the † ‡ B. L. SANZ RESSEL*, , A. R. MASSONE and C. G. antimicrobial resistance of SP strains isolated from † BARBEITO*, dogs with pyoderma and otitis externa (OE). The rela- *Laboratorio de Histologıa y Embriologıa Descriptiva, tionship between hospital size and MRSP isolation rate Experimental y Comparada (LHYEDEC), Facultad de in Japan was analysed. A total of 95 private clinics and Ciencias Veterinarias (FCV), Universidad Nacional de 2,294 dogs with SP infections between 2017 and 2018 La Plata (UNLP), La Plata, Buenos Aires, Argentina; were selected. SP and meticillin resistance were detected † FCV, Consejo Nacional de Investigaciones Cientıficas y by a multiplex PCR strategy for identifying the ther- Tecnicas (CONICET), UNLP, La Plata, Buenos Aires, monuclease gene and the oxacillin susceptibility test, ‡ Argentina; Laboratorio de Patologıa Especial respectively, according to the Clinical and Laboratory Veterinaria Dr. Bernardo Epstein, FCV, UNLP, La Standards Institute (CLSI) Guideline. MRSP was Plata, Buenos Aires, Argentina detected in 987 dogs (43.0%). The isolation rate of MRSP in large clinics (n = 32) with >10 veterinary Cutaneous squamous cell carcinoma (CSCC) represents practitioners was 51.2% (411/803), which was signifi- the most common malignant tumour of the feline skin. cantly higher (P < 0.01) than that in small clinics Research aimed at clarifying how the deregulated activ- (n = 36) with <2 veterinary practitioners (34.45%, 154/ ity of signalling pathways contributes to CSCC progres- 447) and medium-sized clinics (n = 37) with 3–9 veteri- sion can help to identify suitable molecular targets for nary practitioners (40.4%, 422/1,044). An antibiogram the development of novel therapeutic approaches. In subsequently was constructed based on the periodical this regard, recent evidence indicates that the develop- antimicrobial susceptibility data for 12 antibiotics, ment and progression of human CSCC involve the dys- including oxacillin (MPIPC), clavulanic acid-amoxicillin regulated activation of the phosphatidylinositol 3- (AMPC/CVA), cefalexin (CEX), cefpodoxime (CPDX), kinase/Akt/mammalian target of rapamycin (PI3K/Akt/ enrofloxacin (ERFX), gentamycin (GM), trimethoprim- mTOR) signalling pathway, which is a key regulator of sulfamethoxazole (ST), clindamycin (CLDM), doxycy- cell growth and cell fate. Thus, the present study inves- cline (DOXY), minocycline (MINO), chloramphenicol tigated the immunohistochemical expression pattern of (CP) and fosfomycin (FOM), from one large veterinary relevant signalling proteins involved in the PI3K/Akt/ Tyr1068 clinic in Tokyo, with 11 veterinary practitioners, from mTOR signalling pathway including, pEGFR , Ser473 Ser235/236 2017 to 2018. The antimicrobial susceptibility was pAkt , pS6 combined with Ki-67, and the detected by disk diffusion susceptibility tests in accor- tumour suppressor protein PTEN. These signalling pro- dance with the CLSI Guideline. Fifty-three SP strains teins were investigated in 45 feline CSCC samples using were collected from dogs with pyoderma (n = 30) and a tissue microarray. In normal feline skin controls, all OE (n = 24). The resulting antibiogram depicted low proteins showed a low expression in the epidermis, Tyr1068 Tyr1068 susceptibility rates to MPIPC (38%), CEX (49%), except pEGFR . In CSCC, pEGFR immun- CPDX (34%), ERFX (19%), GM (42%), ST (32%), odetection showed that the persistent activation of this CLDM (17%), DOXY (15%) and MINO (23%), and receptor was a frequent event in 31 of 45 samples, moderate susceptibility rates to AMPC/CVA (66%), showing membrane and cytoplasmic immunoreactivity CP (51%) and FOM (58%). Based on these results, the in the epidermal cells of the basal and spinous strata. Ser473 hospital introduced strict antimicrobial selection criteria Likewise, pAkt was overexpressed in 30 of 45 from 2018 for treating dogs with pyoderma and OE, CSCC samples, showing cytoplasmic and nuclear which included: (i) fluoroquinolones, first and third immunoreactivity in the basal and suprabasal cell generation cephalosporins for systemic treatment were strata. Forty-one of 45 CSCC samples also showed a Ser235/236 restricted, (ii) CP and FOM could be used for empirical high epidermal expression for pS6 , with many treatment, and (iii) ST, CLDM, DOXY and MINO of the cells having cytoplasmic staining located in the

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 22 Abstracts basal and parabasal cells strata. Indeed, the double (Patient B) was reported to the author. The veterinar- immunostaining of pS6Ser235/236 combined with the pro- ian and owner were contacted and permitted release of liferation marker Ki-67 showed the co-expression of Patient B’s medical record for study. It was discovered these molecules in some basal and parabasal neoplastic that Patient B was obtained from the same shelter cells of most CSCC samples, a finding that was not approximately one year before Patient A and both observed in normal feline skin. We also have identified patients originated from Rock Springs, Wyoming, a group of CSCC feline patients displaying active Akt USA. Patient B’s record described 13 documented and S6 in the absence of EGFR activation. Expression wounds of the ear, neck, elbow or shoulder over a per- of PTEN characterized by a cytoplasmic immunoreac- iod of 28 months, starting at approximately 5 months tivity in the basal and spinous cell strata was reduced of age. Ulcers were reported to occur after play or epi- or absent in 15 of 45 CSCC samples. These findings sodes of scratching. Wounds continued despite rear show that the persistent activation of the PI3K/Akt/ declaw. Additionally, a Grade 3 patellar luxation was mTOR signalling pathway represents one of the key observed at approximately 18 months of age. Ethylene- events during CSCC progression in cats, pointing to diaminetetraacetic acid blood samples were obtained this signalling pathway as a potential therapeutic target and genome sequencing was performed in both in CSCC feline patients. These results also demonstrate patients. A non-sense variant in the COL5A1 gene was that the activation of the PI3K/Akt/mTOR signalling identified, XM_023242950.1:c.3514A>Tor pathway is independent from EGFR activation in some XP_023098718.1:p.Lys1172*. Sanger sequencing con- feline CSCC, suggesting that therapeutic approaches firmed that both patients were heterozygous at this aimed at modulating the growth and survival of non-sense variant, indicating a common ancestor of the tumour cells by the use of EGFR inhibitors would be two cats and an autosomal dominant mode of inheri- ineffective as sole therapeutic agents in some CSCC tance. The finding of a COL5A1 variant with autoso- feline patients. mal dominant inheritance as well as the array of Source of funding: Self-funded. clinical signs displayed suggest that cats may be a suit- Conflict of interest: None declared. able clinical model of Classical EDS in humans. Source of funding: Self-funded. Conflict of interest: None declared.

SOS-13 A heterozygous novel non-sense variant in the SOS-14 COL5A1 gene in two likely related Bombay cats with Ehlers–Danlos syndrome Onset and duration of action of lokivetmab at 2 mg/kg in a canine model of IL-31-induced J. SCHISSLER*, F. LEUTHARD†,‡,V. JAGANNATHAN†,‡ and T. LEEB†,‡ pruritus *Department of Clinical Sciences, Colorado State A. GONZALES, T. FLECK, L. NORRIS, S. University College of Veterinary Medicine and MAHABIR, S. DUNHAM and O. MARTINON Biomedical Sciences, Fort Collins, CO, USA; †Institute Veterinary Medicine Research & Development, Zoetis, of Genetics, Vetsuisse Faculty, University of Bern, Bern, Inc., Kalamazoo, MI, USA ‡ Switzerland; DermFocus, University of Bern, Bern, Lokivetmab (Cytopoint) is a caninized monoclonal Switzerland antibody that specifically targets canine interleukin Ehlers–Danlos syndrome (EDS) in humans is classified (IL)-31 and is approved for dogs with allergic and ato- into 13 subtypes involving not only cutaneous asthenia, pic dermatitis. Lokivetmab functions to block IL-31- but also musculoskeletal, vascular, neurological and mediated cell signalling, providing relief from condi- mast cell activation disorders. EDS is a result of colla- tions associated with allergic dermatitis such as pruritus gen and/or extracellular matrix changes and its preva- and skin inflammation. Two studies utilized an estab- lence in people is estimated at 1:2,500–5,000 lished, well-characterized canine model of IL-31- individuals. The most common form, classical EDS, is induced pruritus in which pruritic behaviours such as typically caused by COL5A1 gene variants resulting in licking, scratching, head-shaking and body rubbing are abnormal collagen type V. In cats, presumptive EDS observed by video surveillance (in real time) and scored with cutaneous laxity has been rarely described, with over a 2 h period following injection of purified canine variable concurrent vasculopathy, joint laxity and epi- IL-31. In both studies, 24 laboratory beagles ranging in lepsy. An 8-month-old female spayed Bombay cat age from 1.5 to 4.7 years of age (n = 6/group) were (Patient A) presented for lumbar self-trauma, periaural evaluated for pruritus. Categorical presence/absence pruritus, and four wounds of the neck and lumbar (yes/no) of pruritic behaviour displayed by each animal region since adoption from a shelter four months previ- was continuously measured via real-time video surveil- ously. Histopathological evaluation indicated severe lance in discrete 1 min periods for 2 h and hypotheses leucocytoclastic vasculitis and superficial collagen calci- were tested at the two-sided 10% significance level. The fication. Videos demonstrated cutaneous trunci spasms first study objective was to evaluate the duration of with frantic licking and biting of the dorsum. Exam antipruritic activity of multiple dose levels of lokivet- findings included a skin extensibility index of 19%, mab. Subcutaneous administration of placebo or loki- focal alopecia and papulocrustous dermatitis of the vetmab (0.125, 0.5 or 2.0 mg/kg) occurred on study day base of the neck/top of head, and a 7.5 cm full-thick- (D)0. Pruritic activity of dogs then was evaluated in the ness lumbar ulcer. Three months after presentation, morning on D14, D28, D42 and D56. On those days, Patient A experienced multiple partial and grand mal dogs were given a 1.75 µg/kg intravenous bolus of seizures necessitating phenobarbital therapy. A second canine IL-31 approximately 15–20 min before the female spayed Bombay cat diagnosed with EDS scheduled 2 h observation period. The duration of

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 Abstracts 23 action was defined by a statistically significant reduc- are present in the serum of allergen-sensitized humans, tion in pruritic behaviour relative to placebo. Study dogs and cats, and these CCD-specific antibodies might results showed that a significant reduction in pruritus yield false positive results in serological tests for detec- was observed up to D14, D28 and D42 for doses of tion of allergen-specific immunoglobulin (Ig)E. The 0.125, 0.5 and 2.0 mg/kg, respectively. Statistical analy- objective of this study was to determine whether or not sis of the data with mean pruritic scores and corre- similar CCD-reactive antibodies occur in horses, and to sponding P-values relative to placebo for D 14 (91 investigate the prevalence of CCD-reactive IgE antibod- versus 52, P < 0.001), D28 (81 versus 55, P = 0.0218) ies in equine sera using a previously defined inhibitor and D42 (89 versus 60, P = 0.0257) support the (BROM-CCD) and a monoclonal cocktail based antipruritic properties of lokivetmab. In order to evalu- enzyme-linked immunosorbent assay designed to detect ate the onset of action of lokivetmab, laboratory bea- allergen-specific IgE in horses. Sera from 28 horses, gles were given 2.0 mg/kg lokivetmab s.c., then IL-31 presumed to be allergic, were evaluated with and with- was administered 2.5 h post-lokivetmab administration, out BROM-CCD using a panel of 72 different allergens and dogs were evaluated for pruritus between 3 and that included 15 grasses, 17 trees, nine weeds, five 5 h post-treatment. These results demonstrate that a mites, 12 fungi, 12 insects and two environmental aller- single subcutaneous 2 mg/kg dose of lokivetmab signifi- gens. Without BROM-CCD, 22 horses reacted to at cantly reduces pruritic activity in a canine model of IL- least one of the allergens in the panel. Of these reactive 31-induced pruritus when efficacy is evaluated 3–4h horses, 15 horses were reactive to ≥10 allergens, and (P < 0.0001, mean pruritic score for placebo = 37 ver- nine of these had demonstrable IgE to >25 of the aller- sus 10), 4–5h (P < 0.0001, 52 versus 20) or 3–5h gens. BROM-CCD had little or no effect on the mite (P < 0.0001, 90 versus 29) post-antibody administration reactivity in any of the positive samples; however, sub- and the duration of antipruritic activity was sustained stantial inhibition for pollen allergens (trees, grasses for ≥42 days. and weeds) was demonstrable. Complete inhibition of Source of funding: Zoetis Inc., Parsippany, NJ, USA. signal was evident in only two of the reactive samples. Conflict of interest: All authors received reimbursements, However, reduction in signal of ≥20% was evident in fees, funding or salary from Zoetis Inc. 20 of the reactive samples and reduction >50% was observed in eight of the reactive horses. Forty percent of the signals evident in 18 of the samples were inhibited by CCD-BROM. These results demonstrate that SOS-15 CCD-reactive IgE antibodies are evident in horses and that BROM-CCD is effective in reducing reactions with Detection and inhibition of IgE antibodies these irrelevant carbohydrates. Thus, inhibition of reactive with cross-reactive carbohydrate CCD reactivity will likely yield a more accurate in vitro determinants in an ELISA for allergen-specific allergen reactivity profile to be used in the selection of IgE in horses allergens for an immunotherapeutic regime. Source of funding: Funding for this study was provided K. ENCK*, K. LEE*, B. MCKINNEY*, K. † by Stallergenes Greer. BLANKENSHIP* and C. MONTESANO Conflict of Interest: KE, KL, BMcK and KB are employ- *Stallergenes Greer, Lenoir, NC, USA; †Montesano & ees at Stallergenes Greer. Tallarico, D.V.M., LLP, New York, NY, USA Recent findings demonstrate that IgE antibodies specific for cross-reactive carbohydrate determinants (CCD)

FC-1 Funded by a grant from ACVD. Comparison of skin thickness and hair follicle Conflicts of interest: None declared. density in horses with pituitary pars intermedia dysfunction (PPID) and aged non-PPID horses FC-2 A. PETERSEN*, S. FRENCH†, A. HASBACH* and Clinical features of hair cycle arrest: a ‡ H. SCHOTT II retrospective study of 63 cases in Japan *Department of Small Animal Clinical Sciences, Veterinary Medical Center, College of Veterinary K. MORI, Y. OSHIMA-TERADA and N. Medicine, Michigan State University, East Lansing, MI, MURAYAMA USA; †Veterinary Diagnostic Laboratory, College of Dermatology Services for Dogs and Cats, Tokyo, Japan Veterinary Medicine, Michigan State University, East Hair cycle arrest (HCA) is a common disease yet there Lansing MI, USA; ‡Department of Large Animal are few retrospective studies. This study aimed to assess Clinical Sciences, Veterinary Medical Center, College of the clinical features of and therapeutic outcomes for Veterinary Medicine, Michigan State University, East HCA in Japan. Sixty-three dogs with HCA were Lansing, MI, USA recruited based on the clinical, endocrinological and There is a paucity of reports describing histopathologi- histopathological features. The therapeutic evaluation cal characterization of equine skin. A recent study including four sites (neck, dorsum, ventral region and reported that horses with PPID have thinner neck skin caudal portion) before and after a therapy used a hair – – – as compared to aged horses without PPID. Following regrowth scale (0, 0 25%; 1, 25 50%; 2, 50 75%; 3, – euthanasia (late autumn), we collected skin biopsies 75 100%) based on clinical records and digital images. from nine different locations in PPID horses (n = 9) Treatment success was defined as improvement of more and aged non-PPID horses (n = 5). Areas included than four scores before and after a therapy. Toy poo- proximal and distal dorsal forelimb, mane, tail, fore- dles (30) and Pomeranians (22) were significantly over- < head, side of neck, jugular groove, ventral abdomen represented (P 0.01). Forty-seven dogs were assessed and rump. Biopsies were processed routinely; sagittal for therapeutic outcomes and 48.9% of these showed and transverse (at the level of the sebaceous duct paral- treatment success. Oral tocopherol nicotinate (Juvela lel to the skin surface) plane sections were examined. N, Eisai; Tokyo, Japan) with L-cysteine (HYTHIOL, Skin thickness was measured and the number of hair Hisamitsu Pharmaceutical; Tokyo, Japan), oral osater- follicles were counted in a 2 x 2 mm square superim- one acetate (UROACE, Aska Animal Health; Tokyo, posed on photomicrographs of x4 microscopic magnifi- Japan), oral essential fatty acids (Antinol, V and P; cation of haematoxylin & eosin stained sides. No Tokyo, Japan) and a prescription diet (Derm Defence, significant differences in skin thickness or hair follicle Hill’s-Colgate; Tokyo, Japan) were used to treat; suc- density were found between PPID samples and aged cess rates were 39.5% (17/43), 27.3% (6/22), 18.2% (2/ non-PPID horses. Consequently, data for both groups 11) and 37.5% (3/8), respectively. The median time to – were combined for comparison of skin thickness and treatment success was 148 days (63 1,239 days). The < – hair follicle density between the nine body regions. duration-by-treatment success rates for 3 months, 3 6 – – > Mane and tail skin were significantly thicker than the months, 6 9 months, 9 12 months and 12 months other seven sites. In addition, hair follicle density was were 13.0% (3/23), 47.8% (11/23), 8.7% (2/23), 4.3% greater in the face than other body sites. In conclusion, (1/23) and 26.1% (6/23), respectively. This study indi- we could not replicate a previous finding of thinner cated that oral tocopherol nicotinate and L-cysteine was neck skin of PPID horses versus aged non-PPID the most effective therapy, with 60.8% treatment suc- horses. Nevertheless, we were able to show that skin cess by six months and 26.1% with hair regrowth after thickness is greater in mane and tail as compared to 12 months. the other sites. Additionally, density of hair follicles is Source of funding: Self-funded. greater in the face than other body sites. Conflict of interest: None declared.

FC-3 FC-4 Clinical, cytological, histopathological and The characterization of the cellular immune immunohistochemical findings of cutaneous environment in equine genital papillomas and non-epitheliotropic lymphoma in a cat: case squamous cell carcinomas report I. PORCELLATO*, M. ORLANDI*, P. MODESTO‡, † † V. CIMADOMO‡, C. BRACHELENTE*, L. M. PONTES*, A. CASTRO , T. SILVA ,H. † PESSOA†, F. TOLEDO‡, D. SOUZA§ and F. MECHELLI*, B. PASSERI , K. CAPPELLI*, S. ¶ MECOCCI*, R. GIALLETTI*, A. GHELARDI§ and SUEIRO ‡ *Department of Veterinary Medicine, UNIBRA, PE, E. RAZZUOLI † *Department of Veterinary Medicine, Universita degli Brazil; Autonomous Veterinary, PE, Brazil; † ‡Department of Veterinary Clinical and Surgical Studies, Studi di Perugia, Perugia, Italia; Department of Medical Veterinary Sciences, University of Parma, Federal University of Minas, Gerais, MG, Brazil; § §Department of Veterinary Medicine, UNIBRA, PE, Parma, Italy; Azienda Usl Toscana Nord-Ovest, UOC ¶ Ostetricia e Ginecologia, Ospedale Apuane, Massa, Brazil; Faculty of Agricultural and Veterinary Sciences, ‡ UNESP, SP, Brazil Italy; Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta, Genova, Italy Cutaneous manifestation of primary lymphoma of the skin in cats is a rare condition. This report describes a Equine genital squamous cell carcinomas (SCCs) are case of primary non-epitheliotropic cutaneous lym- common skin tumours, often caused by Equus caballus phoma in an 8-year-old mixed breed cat presenting with papillomavirus 2 (EcPV-2) infection, characterized by multifocal and nodular skin lesions not adhered to deep high local invasiveness and a 20% risk of metastasis to tissues. Some nodules were erythematous and hairless. lymph nodes. Growing evidence, both in humans and Cytological findings from fine needle aspirates revealed animals, indicates that presence and type of immune a high population of atypical lymphoid cells character- infiltrate within the tumour microenvironment can dee- ized by scant to moderate amounts of basophilic cyto- ply influence the behaviour of cancer. As no data on plasm, round and eccentric nucleus, displaying intense the immune environment of genital equine SCCs have anisocytosis and anisokaryosis, little mitosis and abun- been reported, our aim was to characterize the cellular dant lymphoglandular bodies in the background. Histo- immune infiltrate in this tumour. Twenty-five formalin- logically, tumours were replacing subcutaneous tissue fixed, paraffin-embedded (FFPE) cases of equine geni- and were composed by a substantial monomorphic tal SCCs were retrospectively retrieved from our population of large lymphocytes arranged in mantle or archives and submitted for histological evaluation of solid pattern. The cells had scant to moderate amounts inflammatory cells. Real-time PCR was performed on of eosinophilic cytoplasm and rounded cleaved nuclei FFPE samples to assess the presence of EcPV-2 within with prominent small central nucleoli. The cells had the lesions. Immunohistochemical analysis for CD3, intense anisocytosis and anisokaryosis. There generally CD20, MUM1, MAC387, IBA1 and FoxP3 was per- were eight to nine mitosis per high power field (x400). formed. Results confirmed the presence of EcPV-2 in Epitheliotropism was not seen and the neoplasia was all tested cases and showed in most of the cases a mas- + restricted to the subcutaneous tissue. The neoplastic sive plasmacytic immune infiltrate (MUM1 ), with + + cells immunoexpressed CD79a and MUM1 and did not fewer B (CD20 ) and T (CD3 ) lymphocytes. Numer- + express CD3, Tryptase, PAX5 or C-Kit. There was pos- ous histiocytes (IBA1 ), mostly located within the itive proliferation marker in approximately 70% of tumour stroma and also infiltrating nests of neoplastic neoplastic cells. The diagnosis of non-epitheliotropic cells, were present with fewer macrophages and neu- + lymphoma of large cells was made. Diagnosis of pri- trophils expressing calprotectin (MAC387 ). Neu- mary cutaneous non-epitheliotropic lymphoma may be trophils were abundant in ulcerated lesions, whereas in based on cytological and histological findings, yet some cases eosinophils were predominant. The immune molecular diagnosis is the gold standard for treatment, infiltrate observed in equine genital SCCs shows some although prognosis was very poor for this case. similarities with human HPV-induced cervical cancer. Source of funding: Self-funded. Moreover, the presence of plasma cells and B cells, + + Conflict of interest: None declared. IBA1 and FoxP3 cells, may suggest the presence of a tumour-promoting and actively immunosuppressive environment in these tumours. Source of funding: Ministry of Health (Ministero della Salute). Conflict of interest: None declared.

FC-5 FC-6 Severe eosinophilic dermatitis in Jersey cows Development of a quantitative PCR to infested with Leptotrombidium spp. (Acari: evaluate accurately the treatment response of Trombiculidae) dogs with demodicosis † ‡ ,† ‡ ‡ M. C. CADIERGUES*, , J. P. MOULIN ,M.N. S. MARTINEZ* , N. SASTRE , O. FRANCINO ,L. § § ¶ ,† LUCAS , P. DORCHIES* and F. SCHELCHER , MATE*, S. SANCHEZ BRUNI*, M. J. DEL SOLE* § *Small Animal Clinic, Universite de Toulouse, ENVT, and L. FERRER Toulouse, France; †UDEAR, Universite de Toulouse, *Centro de Investigacion Veterinaria Tandil INSERM, ENVT, Toulouse, France; ‡Clinique (CIVETAN), CIC-CONICET-UNCPBA, Tandil, † vet erinaire, Saint-Cer e, France; §Large Animal Clinic, Buenos Aires, Argentina; Hospital Escuela de Pequenos~ Universite de Toulouse, ENVT, Toulouse, France; Animales (HEPA), Facultad de Ciencias Veterinarias de ¶IHAP, Universite de Toulouse, INRA, ENVT, la Universidad Nacional del Centro de la Provincia de Toulouse, France Buenos Aires (FCV-UNCPBA), Tandil, Buenos Aires, ‡ A serious outbreak of pruritic dermatitis was investi- Argentina; Servei Veterinari de Genetica Molecular, Universitat Autonoma de Barcelona, Bellaterra, gated in mid-October in an organic farming dairy herd § with 21 Jerseys, three Bretonne pie noir and four Barcelona, Spain; Departament de Medicina i Cirurgia crossed Abondance–Montbeliarde cows in south west Animals, Universitat Autonoma de Barcelona, Bellaterra, France. Lactating cows (17) had permanent grazing Barcelona, Spain access except for indoor milking times. Five 2-year-old Canine demodicosis is the result of an overpopulation and six 1-year-old heifers had been permanently kept in of Demodex mites, normal inhabitants of most skin of two different meadows, with no contact between them mammals. There is no reliable method to quantify the or with adults, since the beginning of the summer. No number of Demodex mites present in the skin to pro- antiparasitic treatment had been administered for the vide accurate information about treatment response. previous five months. Severe and extensive lesions of The objective of this study was to develop a molecular alopecia, lichenification and crusting were observed on technique to quantify the Demodex load in canine skin the jaws, dewlap, face, perineum, above the udder and in order to evaluate the treatment response in dogs with distal limbs, associated with moderate pruritus. Only demodicosis. For this purpose, a real-time quantitative Jersey cows were affected (10 of 13 lactating cows and (q)PCR was developed using primers that amplified a all three 2-year-old heifers) with various degrees of fragment of the D. canis 18S rRNA gene. Ten-fold dilu- lesions severity. Heifers displayed more intense lesions tions of the DNA extracted from isolated mites were than adults. Microscopic examination of deep skin used to elaborate the standard curve. In addition, 44 scrapes showed numerous specimens of Trombiculidae skin biopsies (4 mm) obtained from nine canine cadav- larvae identified as Leptotrombidium spp. and no other ers and 18 skin biopsies from seven dogs with demodi- ectoparasites. Histopathological examination of skin cosis in day (D)0 and D14, D35 and D56 post- biopsy specimens included severe hyperplastic, superfi- treatment (Animal Welfare Committee, FCV- cial and deep perivascular, oedematous, eosinophilic UNCPBA) were used to assess the number of mites in dermatitis. Complete blood counts revealed mild to the skin of healthy dogs and of dogs with demodicosis. moderate eosinophilia. The final clinicopathological The skin biopsies of healthy dogs were negative in 38/ diagnosis was eosinophilic dermatitis suspected to be a 44 samples and positive (>one mite) in the rest (6/44). consequence of Trombiculidae bites. The lesions com- However, in skin biopsies of diseased dogs the initial pletely self-resolved by the end of December. This case Demodex load was mostly >100 mites, decreasing to report emphasizes the need to include Trombiculidae in 50–100, 10–50 and 0–10 mites at D14, D35 and D56 the differential diagnosis of bovine pruritic dermatoses. post-treatment, respectively. The homology between the Finding lesions only in Jersey cows remained unex- qPCR product and the 18S rRNA gene was confirmed plained and could be hypothetically consistent with by DNA sequencing (GenBank KC010485 – D. canis). individual genetic susceptibility to develop hypersensi- This simple technique could be a useful tool to evaluate tivity reactions, or other predisposing breed factors accurately the response to treatment of dogs with (e.g. feeding habits or skin characteristics). demodicosis. Source of funding: Self-funded. Source of funding: PICT 2016-0608, ANPCyT, Argen- Conflict of interest: None declared. tina (SSB) and HEPA, FCV, UNCPBA (MJDS). Published in Veterinary Dermatology, https://doi.org/10. Conflict of interest: None declared. 1111.vde.12895

FC-7 bodied Demodex mites. The histopathological findings on the dorsum revealed orthokeratotic hyperkeratosis A case report of Demodex cyonis in a toy on the epidermis and the hair follicles, sebaceous gland poodle hyperplasia and apocrine gland dilution. The hair follicles contained Demodex mites. However, there were no K. MORI*, T. MORITA†, Y. OSHIMA-TERADA*, I. apparent inflammatory cells in the epidermis, dermis TODA‡, H. KONDO§ and N. MURAYAMA* and hair follicles. Microscopic evaluation for the Demo- *Dermatology Services for Dogs and Cats, Tokyo, dex mites by transmission electron microscopy? identi- Japan; †School of Veterinary Medicine, Nippon fied the typical opisthosomal organ of male D. cyonis. Veterinary and Life Science University, Tokyo, Japan; The diagnosis based on the clinical and examination ‡Toda Animal Hospital, Tokyo, Japan; §Nihon findings was canine demodicosis caused by D. cyonis. University College of Bioresource Sciences, Kanagawa, Oral sarolaner (Simparica, Zoetis Japan) at 10 mg total Japan dose was administered, and the administration of oclac- Several species of Demodex mites parasitizing dogs, itinib was stopped. One month later, pruritus disap- D. canis, D. injai, D. cornei and D. cyonis have been peared and Demodex mites were not detected by hair described morphologically. Demodex cyonis has been examination or skin scraping. Pruritus immediately reported but without details of the clinical course. This improved with administration of oral sarolaner and report describes the clinical course in a toy poodle with thus D. cyonis could be associated with pruritus. D. cyonis. A 15-year-old, intact female toy poodle pre- Although this and previous reports suggest that D. cy- sented with pruritus, erythema and seborrhoea on the onis might favour sebum, more cases will be needed to dorsum one year previously. The dog had been treated assess predisposing factors. with oral oclacitinib (Apoquel, Zoetis Japan; Tokyo, Source of funding: Self-funded. Japan) at 0.5 mg/kg twice daily for one year. Hair Conflict of interest: None declared. examination and skin scraping showed short- and wide-

FC-8 efficacy of a treatment focused on a unique antibacterial approach in a two month protocol. Bacterial isola- Partial traction/avulsion followed by systemic tion and antibiogram were performed at day (D)0 and and topical glucocorticoid therapy for the D60. Magnetic resonance imaging also was evaluated. management of feline aural inflammatory The protocol was to flush the external ear and, in the polyps case of media otitis, the middle ear; no glucocorticoids † † were used. Medical treatment for otitis externa (OE) J. BACHTEL*, J. SCHISSLER and R. ROSYCHUK included local instillation of gentamycin (G4, Virbac; *Veterinary Referral Center of Colorado, Englewood, Carros, France) used off label and EpiOtic (Virbac) CO, USA; †Department of Clinical Sciences, College of every day. For otitis media, treatment included mar- Veterinary Medicine and Biomedical Sciences, Colorado bofloxacin (Marbocyl, Vetoquinol) topically (off label) State University, Fort Collins, CO, USA and orally and topical EpiOtic. Meloxicam (Metacam, Traction/avulsion therapy for aural polyps in cats has a Boehringer Ingelheim) was used to control pain. Twelve reported 33–85.7% recurrence, likely due to re-expan- dogs representing seven breeds were treated; most were sion of the residual polyp in the middle ear. Post-proce- cocker spaniels (four) and German shepherd dogs dural glucocorticoid therapy may facilitate residual (three). The median age was 6.4 years. Otitis was uni- polyp regression. The efficacy and recurrence of partial lateral in seven dogs and bilateral in five. There were traction/avulsion followed by systemic and topical glu- nine OE cases and three individuals with otitis media. cocorticoid therapy for polyps perforating the tympanic Pseudomonas aeruginosa isolates were sensitive to gen- membrane (TM) and systemic and topical glucocorti- tamycin in 5/12 dogs and sensitive to marbofloxacin in coid therapy for nonperforating polyps were evaluated 7/12. After two months, six dogs were free of Pseu- for 25 aural polyps in 22 cats. Three of 22 cats had domonas (four without otitis, two with other organ- bilateral polyps, each with a perforating and nonperfo- isms), while two dogs still had Pseudomonas otitis. rating polyp. Five cats had only a nonperforating Three dogs were excluded owing to excessive pain (two) polyp. All but one (16/25) perforating polyps had all or deviation from the protocol (one); one dog was lost polypoid material removed from the horizontal canal to follow-up. This study shows the improvement by traction/avulsion (biopsy forceps) to the level of or obtained with a unique antibacterial approach. just within the TM. Other than for biopsy, cats with Source of funding: Study funded by Virbac Corporate, nonperforating polyps did not have polyp material France. removed from the middle ear. Post procedure, 21/22 Conflict of interest: None declared. cats (24/25 polyp ears) were treated with oral prednisolone at a mean dose of 2.37 mg/kg (range of 0.91- 3.2 mg/kg) and tapered over 6–18 weeks (mean 8.72 weeks). One cat received subcutaneous dexamethasone. FC-10 Polyp ears received topical glucocorticoid therapy throughout systemic glucocorticoid treatment. Follow- Bacterial chronic otitis: correlation between up ranged from 95 days to 10.4 years (mean 3.7 years). cytological inflammatory patterns, clinical No recurrence of perforating polyps and no recurrence/ aspect and bacteriology in 64 dogs progression was noted for the nonperforating polyps. V. BRUET, T. BREMENT, M. RIVALAIN and P. Based on these results, partial avulsion/traction fol- BOURDEAU lowed by an eight-week course of prednisolone starting Dermatology, Parasitology, Mycology Service, ONIRIS, at 2–3 mg/kg/day with concurrent topical corticosteroid Nantes, France therapy is as effective as aggressive surgical removal (ventral bulla osteotomy, ear canal ablation, middle ear The objective of this study was to evaluate cytological curettage). patterns in chronic bacterial otitis to characterize their Source of funding: Self-funded. clinical or bacteriological implications. One hundred Conflict of Interest: None declared. and four ears (64 dogs) with chronic (minimum two weeks) bacterial otitis externa (bacteria observed then identified bacteriologically) were included. Primary factors, chronicity, previous topical antibacterial treat- FC-9 ments, pain, clinical scoring (OTIS-3) and classification [erythematous (Er), ceruminous (Cer), erythemato-ceru- Efficacy of antibiotic treatment in minous (ErCer), suppurative (Sup)] were collected. Oti- Pseudomonas aeruginosa otitis in dogs: tis was classified on inflammatory pattern from prospective study in 12 dogs cytological results: bacterial overgrowth (no inflamma- † tory cells) (BOG) (43 ears, 41.4%), suppurative (>90% V. BRUET*, T. BREMENT*, M. FUSELIER and P. neutrophils) (Pyo) (55 ears, 52.9%), pyogranulomatous BOURDEAU* (>50% neutrophils and macrophages) (PyoG) (six ears, *Dermatology, Parasitology, Mycology Service, Oniris, 5.8%). Chi-squared test or Fisher’s exact test were used Nantes, France; †Diagnostic Imaging Service, Oniris, (P < 0.05). No correlation was observed between the Nantes, France cytological pattern and chronicity (over two months: Recent literature on medical management of Pseu- 76% BOG, 85% Pyo, 100% PyoG), pain, OTIS-3 or domonas otitis and the impact of different approaches the presence of Malassezia. There was a statistical cor- (antibacterial, glucocorticoid) on healing are lacking. In relation between cytological pattern and: primary fac- this prospective study, the aim was to evaluate the tors (BOG observed in 89.3% of allergic otitis)

(P=0.02); number of previous antibacterial treat- Synelvia. OF is employed by Vetoquinol. FM, JM, JB, ments [0 (58% BOG, 31% Pyo, 17% PyoG) versus 1, 2 CP, JK and DC declare no conflicts of interest. or 3 (P=4.6 9 10-2)]; and clinical type of otitis [BOG with ErCer (61.9%), Pyo and PyoG with Sup (59.2% and 66.7%, respectively)] (P=5 9 10-3). There also was a statistical correlation with bacteria isolated: Sta- FC-12 phylococcus and Corynebacterium with BOG (53.8% and 50%, respectively), Pseudomonas with Pyo/PyoG Florfenicol in vitro antibacterial activity and (80%) (P=0.04); abundance of Malassezia [moderate, genetic resistance of Staphylococcus high level with BOG (84.6%)] (P=2.4 9 10–3). These pseudintermedius strains selected from dogs results highlight the importance of cytological inflam- with otitis matory patterns for describing otitis and possible relationships with bacterial causal agent. A. CORONA, A. VERCELLI, L. GUIDI and L. CORNEGLIANI Source of funding: Study funded by Virbac Corporate, Clinica Veterinaria Citta Di Torino, Italy France. Conflict of interest: None declared. Florfenicol (FFC) is a broad-spectrum fluorinated antibiotic, derived from chloramphenicol (Cf) and active against Gram-positive and -negative bacteria. The aims of this study were to describe the in vitro FC-11 antibacterial activity of FFC against Staphylococcus pseudintermedius (SP) strains and to evaluate the pres- Clinical and microbiological performances and ence of resistance genes. Six hundred SP strains from effects on lipid and cytokine production of a ear canals of dogs with otitis were identified using API cerumenolytic ear cleaner in canine (bioMerieux; Marcy-l’Etoile, France). In vitro suscepti- erythemato-ceruminous otitis bility was tested with the “minimum inhibitory concentration (MIC) Test Strip” method (Liofilchem; Roseto F. MOOG*, J. MIVIELLE*, J. BRUN*, N. degli Abruzzi, Italy) with values between 0.016 and † ,‡ AMALRIC , L. A. LECRU*, C. PRESSANTI* ,J. 256 µg/mL. After heat treatment, extracted bacterial ,‡ KONDRATJEVA*, D. COMBARROS* ,O. DNA was resuspended in Tris-HCl-EDTA (TLE) buf- § ,‡ FANTINI and M. C. CADIERGUES* fer and stored at -20°C. Real-time PCR was performed *Small Animal Clinic, Universite de Toulouse, ENVT, with Step One (Life Technologies Corporation, USA) † ‡ Toulouse, France; Synelvia, Labege, France; UDEAR, for identification of the cfr (chloramphenicol/florfenicol Universite de Toulouse, INSERM, ENVT, Toulouse, § resistance) and fexA genes. The reaction included a France; Vetoquinol, Paris, France Fast SYBR Green Master Mix with primers cfr-fw/cf- Erythemato-ceruminous otitis (ECO) is the most com- rv and fexA-fw/fexA-rv. Melting temperature analyses mon type of otitis in dogs and generally is associated and relative curves were performed. The amplification with bacterial and/or fungal infections. Performance of products were tested using a 2% agarose gel elec- the ear cleaner Sonotix (Vetoquinol; Lure, France) was trophoretic run containing ethidium bromide, to show assessed over two weeks in canine ECO, associated with the bands of at 746 and 1,272 bp corresponding to cfr a mild or moderate secondary infection, in a prospec- and fexA, respectively. Of the SP strains, 99.3% had a tive open-label study (Ethics Committee approval no. MIC to florfenicol from 0.064 to 4 µg/mL and 0.7% SSA_2019_003). Forty ear canals with ECO not receiv- showed a MIC of ≥24 µg/mL. The PCR for cfr and ing any type of aural treatment or cleaning for seven flexA genes were positive for 100% of four strains with days were included. Pruritus (PS), OTIS3 and 0–4 scale high MIC (≥24 µg/mL) and corresponded to 0.7% of cytological (CYTO) scores were assessed on Day (D)0, all SP tested, demonstrating an effective inhibiting D7 and D14. Aural surface concentrations of a panel action against SP. However, as resistance genes can be of 13 cytokines and lipid profiles of the exudate were acquired during FFC-based treatments, FFCs should measured on D0 and D14. Ear cleaning was performed be used with caution. daily if the secretion score (SEC) was 3/3, every second Source of funding: Self-funded. day if 2/3 and every third day if 1/3 from D0 to D12 Conflict of interest: None declared. or D13. PS, OTIS3, SEC and CYTO were significantly lower on D7 compared to baseline (–40%, –31%, – 36%, –34%, respectively; P < 0.001). PS, OTIS3, SEC and CYTO further decreased on D14 (–60%, –53%, – FC-13 61%, -73%, respectively; P<0.0001) and Interleukin 8 and chemokine (KC)-like amounts also were reduced Evaluating factors associated with Malassezia compared to baseline (–62%, P < 0.05; –36%, P = 0.3, pachydermatis presence in canine otitis externa respectively). The lipid profile also was modified with A. FLORES*, C. ROMERO† and L. G. B. GOMEZ ‡ reduced free lipids and increased bound lipids. The use *University of the State of Mexico (UAEM), of Sonotix at a frequency based on the amount of aural Amecameca, Mexico; †DERMAVET, Iztapalapa, secretions significantly decreased the infectious and Mexico City, Mexico; ‡University Center-Amecameca inflammatory components of ECO and could provide Unit of the Autonomous University of the State of an additional tool for managing this type of otitis. Mexico UAEM, Amecameca, Mexico Source of funding: Vetoquinol, Lure, France. Factors such as breed, age and air conditioning expo- Conflicts of interest: MCC has previously been a consul- sure influence the development of yeast otitis. The distant for Vetoquinol. The residency programme of LAL ease often is diagnosed by cytological evaluation yet was supported by Vetoquinol. NA is employed by PCR is useful to detect Malassezia pachydermatis. This

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 30 Abstracts study aimed at evaluating the presence of yeast in the spp. was isolated from one ear. Overall, cocci were external ear canal using PCR and cytological methods found from 40 ears (67%) and rods/coccobacilli from in order to establish the correlation between yeast and 33 ears (55%). Malassezia were not found. There was factors associated with canine otitis. A comparative poor correlation of culture and cytological results, with cross-sectional study approved by the ethics committee microbes not apparent in 43 ears (72%) and 95% of of the UAEM was conducted (registration no. 228/ OIF. Cocci were detected in four ears (7%; median 0, CUAMECAMECA / UAEMEX / 2019). Sixty dogs range 1–3/OIF), rods in 12 ears (20%; median 0, range were examined. NL1 and NL4 primers were used in the 1–14/OIF), and yeast in five ears (8%; median 0, range PCR test. Additionally, a 600 bp fragment was 1–2/OIF). By contrast with previous studies, bacterial sequenced. BLAST was used to compare the obtained rods were present in normal feline ear microbiota and DNA sequence and reported sequences. We conducted microbes were sparse. a phylogenetic analysis using MEGA X 10.0 software. Source of funding: Self-funded. We used a chi-squared test (P<0.05) to assess the Conflict of interest: None declared. factors associated with the presence of M. pachydermatis detected by PCR and cytological evaluation. By PCR we determined that the associated factors are previous otitis, itching and air conditioning exposure. FC-15 Cytological results corresponded with predisposing factors of itching, ear shape, hair inside the ear and skin Cytological and microbial culture from 60 health. The obtained sequence was combined at 99.9% normal canine ears with M. pachydermatis. The phylogenetic analysis L. J. VOGELNEST*,†, A. P. KING† and J. M. showed that this sequence is close to sequences reported NORRIS† in Colombia and Kuwait corresponding to a zoonotic *Small Animal Specialist Hospital, North Ryde NSW string endangering owners and veterinarians. Knowing Australia; †School of Veterinary Science, University of the predisposing factors for otitis externa is a tool to Sydney, NSW Australia obtaining the diagnosis. No studies on cytological findings from normal canine Source of funding: Self-funded. ears evaluated with oil-immersion microscopy have Conflict of interest: None declared. been published. This prospective study (University of Sydney ethics N2016/1089) evaluated cytological results and concurrent microbial culture from 60 normal ears (30 dogs). Two sterile swabs were collected; the first FC-14 placed into transport medium and inoculated onto Cytological and microbial culture from 60 sheep blood agar and Sabouraud’s dextrose agar within normal feline ears 10 h, with plates incubated aerobically at 37(C for 48 h. Bacterial isolates were identified using matrix- L. J. VOGELNEST*,†, A. NESSIEM† and J. M. † assisted laser desorption/ionization-time of flight. The NORRIS second swab was rolled onto a glass slide (2 x 2 cm *Small Animal Specialist Hospital, North Ryde, NSW, † area) which was stained with modified Wright’s stain. Australia; School of Veterinary Science, University of Keratinocytes were identified under x40 magnification, Sydney, NSW, Australia and nine oil-immersion fields (OIF; x100 magnification) This prospective study (University of Sydney ethics containing keratinocytes evaluated for microbes. Bacte- N2016/1088) evaluated cytological findings and concur- ria were cultured from 48 ears (80%) with 93 species rent microbial culture from 60 normal ears (30 cats). isolated. Staphylococcus (62%), Bacillus (33%) and Sterile swabs inserted 1–2 cm into external canals were Micrococcus (30%) were the most frequent genera, and rotated for 1–3 s. An initial swab placed into transport S. pseudintermedius, Micrococcus luteus and S. hominis medium was inoculated onto sheep blood agar and the most frequent isolates. Cocci were isolated from 44 Sabouraud’s dextrose agar within 10 h. Plates were ears (73%), rods/coccobacilli from 37 ears (61%), incubated aerobically at 37C for 48 h and bacterial iso- Malassezia from 29 ears (48%), Pseudomonas spp. from lates identified using matrix-assisted laser desorption/ six ears and Aspergillus flavus from one ear. Cytological ionization-time of flight. A second swab was rolled results revealed cocci in 58 ears (97%; median 1, range onto a glass slide (2 x 2 cm area) and stained with 1–8/OIF), rods in 56 ears (93%; median 1, range 1-6/ modified Wright’s stain. Keratinocytes were identified OIF) and yeast in 40 ears (67%; median 5, range 1 to under x40 magnification, and nine oil-immersion fields >100/OIF). Five dogs with >100 yeasts in at least one (OIF; x100 magnification) with keratinocytes were eval- OIF were excluded from median counts. By contrast to uated for microbes. Bacteria were cultured from 50 ears other studies, bacterial rods were identified as part of (83%) with 63 species isolated. Staphylococcus (25%), normal canine ear microbiota and yeasts were plentiful Bacillus (22%) and Lactobacillus (6%) were the most in some normal ears. frequent genera, and S. felis, S. epidermidis and Micro- Source of funding: Self-funded. coccus luteus the most frequent isolates; Pseudomonas Conflict of interest: None declared.

FC-16 FC-17 Carriage of Malassezia yeasts on healthy dogs Comparative study of microbubble wash in Malaysia system and regular shampooing in the effect on A. L. BOO detergency and skin barrier function of dogs Pet First Veterinary Centre, Kuala Lumpur, Malaysia K. SHIMADA* and T. IWASAKI† † Malassezia spp. yeasts are both commensal organisms *Azabujuban Dog and Cat Clinic, Tokyo, Japan; Vet and opportunistic pathogens of canine skin. Although Derm Osaka, Osaka, Japan canine Malassezia dermatitis is a common diagnosis in Microbubble wash system (MWS) is a new technology Malaysia, little is known about their presence on the to wash animal skin by using microbubbles without skin of healthy dogs there. This study was performed scrubbing. The purpose of this study was to evaluate to evaluate the prevalence of carriage and quantifica- the effects of MWS on the canine skin detergency and tion of Malassezia spp. yeasts in healthy dogs in barrier function in comparison with regular shampoo- Malaysia. Thirty-eight clinically healthy dogs admitted ing. B-WASH-ST-W (Thales Inc.; Tokyo, Japan) was for neutering in a small animal veterinary clinic were used as a microbubble generator in conjunction with enrolled in the study. Swabs were collected from bilat- bath salts and enzymes. A shampoo containing no eral ear canals and tape-strip samples were obtained moisturizing components was used as control. Sham- from interdigital skin of both forelimbs. Cytological pooing and MWS were performed on each side of the examination was performed and results were expressed lateral thorax of five mixed-breed dogs with healthy as the mean number of yeasts per x1,000 microscopic skin. The detergency was measured from the amount of field per dog. Fifty percent of the otic samples (n = 74) sebum extracted from the hair coat (per g) with chloro- and 68% of the samples from the interdigital skin form and methanol solutions (2:1) at time (T, min) 0, (n = 38) showed presence of Malassezia spp. yeasts on T5, T10, T15 and T20 of washing. Skin barrier function cytological examination. The mean number of Malasse- was evaluated by measuring transepidermal water loss zia spp. yeasts was 1.86/oil-immersion field (high-power (TEWL) with a closed chamber evaporimeter (CC-01, field, HPF) and 1.19/HPF for the otic samples and Kao Corporation; Tokyo, Japan) at T0, T10 and T20 interdigital skin, respectively. There was no statistically of washing. The detergency of MWS at T15 of washing significant difference in prevalence and number of was equivalent to that of shampooing at T5 (P = 0.855, Malassezia spp. yeasts between dogs of different ages, paired Student’s t-test). At T10 and T20, the TEWL sex, breed and area of sampling (P > 0.05). These after the shampooing treatment was significantly higher results are in accordance with previous findings in the (21.51 Æ 1.36 g/m2/h and 26.13 Æ 2.42 g/m2/h) com- literature which suggested that a clinical diagnosis of pared to after the MWS (17.40 Æ 3.63 g/m2/h and Malassezia dermatitis may be established if >2/HPF 16.62 Æ 2.43 g/m2/h) (P < 0.05, one-factor ANOVA). Malassezia spp. yeasts are found on cytological analysis These observations suggest that MWS can clean the coupled with compatible skin lesions. skin surface and hair coat of dogs without scrubbing Source of funding: Self-funded. and impairs the skin barrier function less than sham- Conflict of interest: None declared. pooing. Source of funding: Self-funded. Conflict of interest: None declared.

FC-18 kg twice daily. Group 2 was induced with oclacitinib at a mean dose of 1 mg/kg twice daily. Mean age of dogs Oclacitinib in the treatment of pemphigus was 6.5 years. Patients were evaluated at four visits on foliaceous in dogs day (D)1, D14, D30 and D60 using a validated lesion A. M. CORDERO*, C. LOPEZ-MARQUEZ*, G. score system. Patients in Group 2 had a decrease of SHEINBERG† and C. ROMERO§ clinical score of 65% and 84.8% at visits 3 and 4, *Vetderm, Guadalajara, Mexico; †Centro Veterinario respectively. Group 1 had a 15% decrease in clinical Mexico, Mexico City, Mexico; §Dermavet, Mexico City, score at visit 4. One patient in Group 1 discontinued Mexico follow-up at the third visit. One dog in Group 2 developed a fibrosarcoma and another was diagnosed with Pemphigus foliaceous (PF) is the most common cutaneous T-cell lymphoma; both were removed from autoimmune skin disease in dogs. Its treatment requires the study at visits 3 and 4. Oclacitinib seems to be an long-term management with immunosuppressive drugs, alternative for maintenance and induction in the treat- often needed life-long. The objective of this study was ment of PF in dogs. Further studies are required to to evaluate the therapeutic effect of Oclacitinib (Apo- evaluate high dose and long-term use of oclacitinib in quel, Zoetis; New Jersey, NY, USA) in the treatment dogs and different treatment combinations to achieve of PF in dogs. Eight dogs diagnosed clinically, cytologi- clinical control or remission for PF. cally and histologically for PF were divided into two Source of funding: Self-funded. groups. Owners signed a consent form. Group 1 was Conflicts of interest: None declared. induced with immunosuppressive doses of prednisone and/or azathioprine in addition to oclacitinib at 1 mg/

FC-19 FC-20 Effects on clinical signs, lipids and hydration Performance of combined shampoo and mousse factors of combined applications of shampoo applications containing ophytrium and and mousse containing ophytrium and seboliance in dogs with scaling disorders: a seboliance in seborrhoeic dogs European field trial J. KONDRATJEVA*, J. BRUN*, N. AMALRIC†,F. M. GATELLET*, S. AHMAN †, V. BRUET‡,P.M. MOOG*, D. COMBARROS*,‡, C. PRESSANTI*,‡,C. CADOT§, R. S. MUELLER¶, C. NOLI††,T. ZEMIRLINE§, N. MAUBERT§, M. GATELLET¶ and NUTTALL‡‡, E. OLLIVIER*, T. BLONDEL* and N. M. C. CADIERGUES*,‡,¶ SAVELLI§§ *Small Animal Clinic, Universite de Toulouse, ENVT, *Ceva Sante Animale, Libourne, France; †VetaDerm Toulouse, France; †Synelvia, Labege, France; ‡UDEAR, Veterinarklinik,€ Lomma, Sweden; ‡SkinVet Universite de Toulouse, INSERM, ENVT, Toulouse, Dermatologie Vet erinaire, Nantes, France; §Clinique France; §Ceva Sante Animale, Louverne, France; ¶Ceva vet erinaire SeineVet, Rouen & Boos, France; ¶Ludwig Sante Animale, Libourne, France Maximilian University, Munich, Germany; ††Servizi ‡‡ Rapid improvement of skin appearance in case of seb- Dermatologici Veterinari, Peveragno, Italy; The Royal orrhoeic disorders is important to bring comfort to the (Dick) School of Veterinary Studies and The Roslin §§ dog and encourage owner compliance. The objective of Institute, Roslin, Scotland; Nadege Savelli EIRL, this study was to evaluate the tolerance, performance Villeneuve Loubet, France and effect on hair lipids and skin hydration of a sham- Besides treating the underlying disease, it is essential to poo and mousse containing plant extracts (ophytrium improve skin appearance and odour in dogs with dry and seboliance, DOUXO S3 SEB, Ceva Sante Animale; or greasy scaling. The objective of this study was to Libourne, France) in seborrhoeic dogs. Eighteen dogs evaluate the performance of the combined application with idiopathic chronic seborrhoea oleosa were enrolled of a shampoo and mousse containing the plant extracts in a prospective controlled study. Six control dogs were ophytrium and seboliance (DOUXO S3 SEB Shampoo washed with plain water on day (D)0. Twelve dogs and Mousse, Ceva Sante Animale; Libourne, France) were shampooed on D0 followed by eight mousse in dogs with scaling disorders. Nineteen dogs with dry applications every 48–72 h from D2 to D18. Clinical or greasy scaling were included in this prospective, mul- score (CS), natural moisturizing factors (NMF) and ticentric, open study. Dogs were shampooed on day hair lipids (HL) were assessed on D0, D0 + 4h, D7, (D)0 and received eight mousse applications every 48- D14 and D24. At baseline, CS, NMF and HL were not 72 h. Skin Seborrhoeic Index (SSI) was assessed by a significantly different between groups and remained dermatologist on D0, D7 and D21. This included stable throughout the study in the control group. A sta- odour, erythema, scaling, greasiness, extent and sec- tistically significant reduction in CS from D0 to D7 (– ondary signs, each scored on a 0–3 scale. On D21, own- 53%) was observed in the test group reaching 91% on ers and veterinarians scored the protocol performance D24 (P < 0.05). No adverse effects were noted. NMF and satisfaction. Mean SSI decreased significantly after and HL levels decreased in the test group on D0+4h (– 21 days (P < 0.05) with a mean 33.9% improvement. 73%, P < 0.0001 and –50%, P = 0.14, respectively) and On D21, the mean odour score decreased by 54.5%, returned to baseline values on D14 and D7, respec- extent by 46.1%, greasiness by 40.3% and scaling by tively. The combination of one shampoo and subse- 33.3%. Most owners (84.2%) were satisfied or very sat- quent mousse applications every 48-72 h containing isfied with the performance of the protocol, emphasiz- ophytrium and seboliance can be recommended as a ing the good appearance of the skin in 89.4% of cases. convenient and safe protocol to quickly improve hair Veterinarians assessed the protocol as satisfactory, coat quality in seborrhoeic dogs without dehydrating good or excellent in 84.2% of cases. One shampoo and the skin. eight mousse applications of the products containing Source of funding: Ceva Sante Animale, Libourne, ophytrium and seboliance improved dry or greasy scal- France. ing in the study dogs. There was a noticeable improve- Conflicts of interest: MCC has previously been consul- ment in some parameters important for pet owners, tant for Ceva Sante Animale. NA is employed by Synel- such as odour, resulting in high owner satisfaction via. CZ, NM, MG are employed by Ceva Sante levels. Animale. JK, JB, FM, DC and CP declare no competing Source of funding: Ceva Sante Animale, Libourne, interests. France. Conflict of interest: All authors received reimbursements, fees, funding or salary from Ceva Sante Animale.

FC-21 the majority from Germany) were subjected to a seasonal in vitro allergy test using Elisa Fc-Ԑ receptor tech- Sensitization to the crude Dermatophagoides nology (Heska; Fribourg, Switzerland). Polysensitized farinae extracts Der f 2 and Zen 1 in dogs samples were re-tested after adding a modified glyco- with atopic dermatitis in southern Brazil protein plant extracts (CCD inhibitor/CHO blocker; L. LUDWIG*, T. TSUKUI†, M. KAGEYAMA† and Heska). Polysensitization was present in 137 (29%) M. FARIAS* samples without a difference by season. After blocking, *Postgraduate Program in Animal Science, School of 48% of the polysensitized sera were negative and 40% Life Sciences, Pontifıcia Universidade Catolica do showed lower antibody levels. Forty-eight randomly Parana, Curitiba, PR, Brazil; †Central Research selected samples were analysed before and after block- Laboratory, ZENOAQ, Koriyama, Fukushima, Japan ing to determine the impact of the CHO blocker on each allergen. The total number of negative reactions Allergens of the mite Dermatophagoides farinae are fre- after the addition of the CHO blocker were higher in quently associated with the sensitization of dogs with nettle (65%), willow (67%), mugwort-ragweed (65%) atopic dermatitis (AD). The aim of this study was to and English plantain (54%) than in a six-grass mix evaluate the sensitization of dogs with AD to the crude (31%), rye (23%) and sheep’s sorrel (25%). The 29% D. farinae extract (cDf), Der f 2 and Zen 1. One hun- prevalence of anti-CCD IgE in cats is similar to that dred dogs of different breeds, ages and sex were reported in dogs and humans. The use of a CHO selected. All dogs had clinical diagnosis of AD and ful- blocker for pollen allergy testing in cats improves the filled at least five of eight pre-established criteria and identification of the relevant offending allergens to for- exclusion of other pruritic skin diseases. Serum samples mulate allergen-specific immunotherapy. of each dog were submitted to ELISA test (ZENOAQ, Source of funding: Self-funded. Koriyama; Fukushima, Japan) for measurement of Conflict of interest: AM, CL, EM, MC and RW work allergen-specific immunoglobulin (Ig)E, with a cut-off for Laboklin. at 0.200 optical density. The results were reported in descriptive data and, for the correlation of each allergen, Spearman’s rank correlation coefficient was carried < out (P 0.05). Of the 100 dogs, 92% were positive to FC-23 cDf, 77% to Zen 1 and 56% to Der f 2. There was correlation between the sensitization to cDf and Zen 1 Prevalence of high levels of specific IgE (Spearman’s r = 0.88, P < 0.001), and not between cDf against mites in dogs and cats from non- and Der f 2 (r = 0.30, P < 0.002), and between Zen 1 Mediterranean Europe = < and Der f 2 (r 0.22, P 0.02). It is concluded that the € sensitization to cDf, Der f 2 and Zen 1 is important in C. LORENTE-MENDEZ, E. MULLER and R. dogs with AD in southern Brazil. WAGNER € Source of founding: The Coordination for the Improve- Laboklin, Labor fur Klinische Diagnostik GMBH & CO. ment of Higher Education Personnel – Brazil (CAPES) KG, Bad Kissingen, Germany and ZENOAQ. Presence of high levels of specific-immunoglobulin (Ig) Conflict of interest: TT and MK are employees of E in serum determines the sensitization of the patient ZENOAQ. to the allergen. The objective of this study was to evaluate the prevalence of high levels of IgE against Alternaria/Aspergillus (A/A) Penicillium/Cladosporium (P/C), Dermatophagoides farinae (Df), Der- FC-22 matophagoides pteronyssinus (Dpt), Tyrophagus putres- centiae (Tp) and Acarus siro (As) in sensitized dogs and Prevalence of IgE against CCD and impact of cats. Veterinarians from non-Mediterranean European CCD blocking in serum from cats countries sent serum samples from cats and dogs to A. MOHAMMADDAVOODI*, C. LORENTE*, L. Laboklin (2017 to 2019) to investigate the allergens PANAKOVA†,EMULLER*,€ M CHRISTIAN* and involved in the allergic reaction of the patient. Sera R. WAGNER* from cats (2,148) and sera from dogs (22,114) were *Laboklin, Labor fur€ Klinische Diagnostik GMBH & analysed using an ELISA test based on the FceR1a CO. KG, Bad Kissingen, Germany; †Clinical Unit of receptor (Heska; Fribourg, Switzerland); 21,261 Internal Medicine Small Animals, Department for (95.70%) of the dog samples and 1,814 (84.45%) of the Companion Animals and Horses, Vetmeduni, Vienna, cat samples yielded positive results to at least one aller- Austria gen. The percentage of high levels of specific IgE in the positive samples of dogs and cats (respectively) were Cross-reactive carbohydrate determinants (CCD) lead 10.66% and 8.88% for A/A, 4.86% and 4.91% for P/ to highly multi-positive results in seasonal in vitro C, 91.10% and 75.91% for Df, 51.98% and 59.81% for allergy tests. False-positive/clinically irrelevant results Dpt, 90.36% and 87.38% for Tp, and 82.23% and arise from the detection of immunoglobulin (Ig)E 64.06% for As. Sensitization to moulds was associated against CCD (anti-CCD IgE). However, there are no with mite sensitization in >99.5% of positive mould studies on this phenomenon in cats. This retrospective samples. Based on these results, moulds seem not to be study aimed to evaluate the prevalence of polysensitiza- an important cause of sensitization in dogs and cats. tion in feline sera and the impact of the use of a CCD Prevalence of sensitivity to all mites was high, most fre- inhibitor (CHO blocker). A total of 472 cat sera (from quently to Df and Tp and least frequently to Dp. veterinary practices in the EU, Switzerland and China,

Storage mites seem to play an important role in the normal primary keratinocyte cell lines in chamber slides allergic reactions of dogs and cats. to study differentiation and tight junction (TJ) expres- Source of funding: Self-funded. sion. Cells were fixed the first confluency day (D)0 and Conflict of interest: All authors work for Laboklin. on D6. Immunofluorescent staining was done for zonula occludens, claudin, occludin, filaggrin and filaggrin-2. Five pictures per chamber for each antibody were taken. Pictures were randomized and scored by FC-24 four blinded observers. Cell size and variability and continuity of TJ proteins were scored. ANOVA showed Prevalence of high levels of specific IgE increased cell size from D0 to D6 in both groups (effect against pollens in dogs and cats from non- of time, P = 0.0076) and no differences between groups. Mediterranean Europe Size was more subject-dependent than disease-related = C. LORENTE-MENDEZ, E. MULLER€ and R. (P 0.0022). No difference for continuity was found WAGNER for any of TJ proteins. Both filaggrins were nuclear Laboklin, Labor fur€ Klinische Diagnostik GMBH & CO. early on and later cytoplasmic. The increase of cyto- KG, Bad Kissingen, Germany plasmic filaggrin-2 on D6 was significant in atopic dogs (P = 0.0085) as was the effect of subject (P = 0.002), Serological allergen tests using the blocking of cross-re- indicating that it was more subject-dependent than dis- active carbohydrate determinants (CCDs) have demon- ease-related. The decrease of staining of filaggrin-2 in strated the same reliability as the intradermal test to the nucleus at D6 approached significance in the atopic identify the allergens involved in pollen-allergic dogs dogs (P = 0.055). It is concluded that the cell culture and cats. This retrospective study aimed to evaluate the behaviour may be linked to factors other than TJ pro- prevalence of high levels of specific immunoglobulin teins. This is the first report of the nuclear transloca- (Ig)E against pollens in hypersensitised dogs and cats tion of filaggrin-2 in dogs. from Europe (non-Mediterranean areas). A total of Source of funding: Self-funded. 10,345 sera from dogs and 1011 from cats were anal- Conflicts of interest: None declared. ysed with an ELISA test based on the FceR1a receptor (Heska; Fribourg, Switzerland) together with CCD inhibition (CHO blocker; Heska) in polysensitized samples. Positive results to at least one allergen were found FC-26 in 8,177 (79.04%) of the dog samples and 750 (74.28%) of the cat samples. The percentage of high levels of Dermatophagoides farinae allergenic profiles specific IgE in the positive samples of dogs and cats in atopic cats and horses: differences between were (respectively) 84.50% and 83.36% for sorrel, species 80.80% and 75.10% for rye, 74.40% and 74.53% for † six-grass mix, 56.80% and 51.53% for English plantain, L. RAMIO-LLUCH*, R. MOYA , A. GONZALEZ- RUIZ†, F. ALVAREZ †, J. CARNES † and A. 36% and 45.54% for willow, 35.10% and 43.54% for ‡ mugwort/ragweed, 27.50% and 36.22% for Birch, and PUIGDEMONT *Animal Health B.U., Laboratorios LETI, S.L.U., 19.90% and 42.08% for nettle. It can be concluded that † in non-Mediterranean Europe, the principal pollen Barcelona, Spain; R&D Immunology and Allergy Unit, Laboratorios LETI, S.L.U., Madrid, Spain; allergens for dogs and cats are sorrel, rye and grass ‡ with similar values of hypersensitivity prevalence Deptartment of Pharmacology, Therapeutics and (>75%) for both species. Plantain is the next most fre- Toxicology, Universitat Autonoma de Barcelona, quent pollen. Willow, mugwort, birch and nettle seem Barcelona, Spain to induce fewer hypersensitivities in dogs and cats, House dust mite (HDM) allergens already have been especially in dogs. reported as an important cause of allergenic reactions Source of funding: Self-funded. in cats and horses, yet there are no studies about the Conflict of interest: All authors work for Laboklin. main proteins involved in the sensitization. The objective of this study was to investigate the allergenic sensitization profile of cats and horses with atopic dermatitis against Dermatophagoides farinae. Nineteen FC-25 cats and 15 horses sensitized to D. farinae [clinical diagnosis confirmed with positive specific immunoglobulin Filaggrins and tight junction proteins (Ig)E levels by ELISA test (Greer; Lenoir, USA)], four expression in keratinocyte cell cultures from control healthy cats and two control healthy horses normal and atopic dogs were included in the study. Allergenic profiles were R. MARSELLA, K. AHRENS, R. WILKES, N. S. investigated by immunoblot. The D. farinae extract MUNGUIA and M. DORR (Laboratorios LETI; Barcelona, Spain) was elec- Department of Small Animal Clinical Sciences, College trophoretically separated by SDS-PAGE, transferred of Veterinary Medicine, University of Florida, and incubated with the serum samples. Membranes Gainesville, FL, USA were incubated with the corresponding secondary antibody (anti-IgE) and developed by chemiluminescence. Keratinocytes of normal and atopic dogs behave differ- Results showed that most cats recognized high molecu- ently in cell culture when creating a monolayer and dif- lar weight (MW) allergens. However, most also showed ferentiating. Normal keratinocytes achieve confluence strong recognition of proteins of approximately first and then differentiate while atopics start differenti- 30 kDa, and only three cats showed recognition of ating, creating little “domes” even if they are not yet lower MW allergens. Feline controls did not show any confluent. In this study we grew four atopic and five band recognition. Different allergenic profiles were

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 36 Abstracts found among positive horses and all sera recognized 37 kDa band. In conclusion, cats recognized mainly allergens of a broad MW range (from 14 to >100 kDa). medium and high MW proteins while horses did not Nonetheless, proteins between 25 and 37 kDa were rec- present a specific HDM sensitization allergenic pattern. ognized for the majority. Horse controls did not show Source of funding: Self-funded. any band recognition except for one control with a Conflict of interest: None declared.

FC-27 inflammatory lesions in atopic dogs, a minority do not show a satisfactory response. To date, the reason for Effectiveness of a nutraceutical with this lack of efficacy in some dogs remains unknown. blackcurrant oil and tyndalised Lactobacillus The objective of this study was to determine if the low reuteri in controlling intestinal dysbiosis and efficacy of the drug observed in these cases could be pruritus in atopic dogs consequence of pharmacokinetic factors. Twenty-four dogs with a diagnosis of atopic dermatitis were treated L. CORNEGLIANI*, P. PERSICO† and A. § with oclacitinib, 0.4–0.6 mg/kg orally twice daily for 14 GRAMENZI days, then once daily for 14 days. Owner-assessed pru- *Clinica Veterinaria Citta Di Torino, Torino, Italy; ritus Visual Analog Scale (pVAS) and clinician-assessed †Private practice, Milan, Italy; §Faculty of Veterinary Canine Atopic Dermatitis Extent and Severity Index, Medicine, University of Teramo, Teramo, Italy 4th iteration (CADESI-04) were evaluated on day (D)0 Canine atopic dermatitis (cAD) is a chronic allergic dis- and D28. The plasma concentration of oclacitinib was ease which requires a multimodal management. As well determined by high-performance liquid chromatogra- as the impaired skin barrier, this disorder causes dys- phy at D28, 1 h after oral administration of oclacitinib. functions of the skin and gut microbiome. The present Levels of oclacitinib on D28 ranged from 1.6 to 476.6 study evaluates the intestinal microbiome, faecal score (average 226.9 Æ 131.5) ng/mL. Correlation was not and Visual Analog Scale (VAS) in dogs affected by observed between plasma concentration of oclacitinib cAD before, during and after the administration of a and reduction in pVAS score (rxy = 0.067, P > 0.05). nutraceutical based on blackcurrant oil, tyndallized Negative correlation was obtained when comparing Lactobacillus reuteri, nucleotides and zinc oxide. Forty- decrease in CADESI-04 score and levels of oclacitinib five dogs with cAD in maintenance therapy for at least (rxy = –0.562, P < 0.05). Plasma concentrations of one year were selected and fed a diet with hydrolysed oclacitinib were highly variable and in most dogs, even chicken and rice throughout the 120 days of the study. in nonresponders, they were in the range considered Throughout the study, current therapies remained therapeutic. These results indicate that the lack of effi- unchanged. Each dog was its own control. From day cacy of oclacitinib in some dogs does not seem to be (D)0 to D60 all dogs were administered the nutraceuti- consequence of pharmacokinetic factors. cal (Ribes Pet Symbio, NBF Lanes; Milan, Italy) once Source of funding: Apoquel tablets were supplied by Zoe- daily, then the administration ceased for the next 60 tis (Parsippany, NJ, USA); the manufacturer was not days. Stool samples were collected on D0, D60 and involved in study design, data collection and interpreta- D120 for microbiome analyses carried out by sequenc- tion or publication. ing the16S rRNA gene with calculation of the dysbiosis Conflict of interest: LF has received unrelated honoraria index and faecal score. VAS was assessed on D0, D60 for lecturing from Zoetis. No conflicts of interest have and D120. Statistical analysis used the mixed-effects lin- been declared by the other authors. ear model. VAS and faecal score decreased significantly between D0 and D60 (P<0.01), while there were no significant variations between D60 and D120. The dysbiosis index decreased (improved) between D0 and D60 FC-29 (P < 0.01) and between D60 and D120 it increased (worsened) significantly (P < 0.01). The study high- Performance of a combined application of lights the usefulness of this nutraceutical in the control ophytrium-containing shampoo and mousse in of intestinal dysbiosis and VAS in atopic dogs. dogs with sensitive skin: a European field trial Source of funding: NBF Lanes, Milan, Italy. † ‡ M. GATELLET*, S. AHMAN , V. BRUET ,P.M. Conflict of interest: NBF Lanes paid the authors to per- § ¶ †† CADOT , R. S. MUELLER , C. NOLI ,T. form this study. NBF Lanes was not involved in the study ‡‡ NUTTALL , E. OLLIVIER*, T. BLONDEL* and N. design or data analysis. §§ SAVELLI *Ceva Sante Animale, Libourne, France; †VetaDerm Veterinarklinik,€ Lomma, Sweden; ‡SkinVet Dermatologie Vet erinaire, Nantes, France; §Clinique FC-28 vet erinaire SeineVet, Rouen & Boos, France; ¶Ludwig Correlation between plasmatic levels of Maximilian University, Munich, Germany; ††Servizi ‡‡ oclacitinib and clinical response in dogs with Dermatologici Veterinari, Peveragno, Italy; The Royal (Dick) School of Veterinary Studies and The Roslin canine atopic dermatitis §§ Institute, Roslin, Scotland; Nadege Savelli EIRL, ,† ,† R. GARCIA-FONTICOBA* , L. ORDEIX* ,M. Villeneuve Loubet, France ,† ,† CABRE* and L. FERRER* Topical products can provide direct relief of the dry *Fundacio Hospital Clınic Veterinari, Universitat and irritated skin in atopic patients. The objective of Autonoma de Barcelona, Barcelona, Spain; †Department this study was to evaluate the performance of a proto- of Animal Medicine and Surgery, Universitat Autonoma col combining a shampoo and a mousse each contain- de Barcelona, Barcelona, Spain ing ophytrium, a purified plant extract with skin Oclacitinib is a selective inhibitor of Janus kinase-1 barrier strengthening, soothing and antibiofilm proper- approved for the treatment of allergic dermatitis in ties in dogs with sensitive skin (DOUXO S3 CALM dogs. Although oclacitinib has been demonstrated as Shampoo and Mousse, Ceva Sante Animale; Libourne, highly effective in controlling pruritus and France). Twenty dogs with a history of atopic

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 38 Abstracts dermatitis presenting a skin flare were enrolled in a nonresponding cat responded to the second ED. Farm- prospective, multicentric, uncontrolled study. Dogs ina UltraHypo seems to be a useful ED when screening were bathed on day (D)0 and then given eight mousse cats for FA. Furthermore, it did not cause reactions in applications every 48–72 h. Canine Atopic Dermatitis cats with rice or fish hypersensitivity. Extent and Severity Index, 4th iteration (CADESI-04) Source of funding: Farmina Pet Food, Nola, Italy. scores were assessed by a dermatologist and pruritus by Conflicts of interest: C.N. has received consultation fee the owners using the pruritus Visual Analog Scale and speakers fee from Farmina. (pVAS) score on D0, D7 and D21. Owner and veterinarian evaluations of performance and satisfaction were recorded on D21. Mean CADESI-04 scores improved throughout the study, from 18 at D0 to 6.4 at D21 FC-31 (58.3% mean improvement; P < 0.05 at all timepoints). On D21, 63.2% of the dogs achieved ≥70% CADESI- Anti-inflammatory effects of oclacitinib on 04 improvement; 78.9% had a score below the target skin lesion scores in a house dust mite-sensitive threshold of 10. Mean pVAS scores decreased signifi- colony of beagles < cantly after 21 days (P 0.05 at all time-points) with J. T. ROSS*, K. RUDOLPH†, J. MLECZKO†,T. mean 30.8% improvement; 42.1% of the dogs had BARRETT† and A. J. GONZALES* ≥ 50% pVAS score decrease. Global veterinary assess- *Global Therapeutics Research, Zoetis, Kalamazoo, MI, ment of the protocol was satisfactory, good or excellent USA; †Lovelace Biomedical, Albuquerque, NM, USA in 73.7% of cases. Most owners (78.9%) were satisfied or very satisfied with the protocol. One shampoo and Oclacitinib is approved for the control/treatment of ≥ eight mousse applications of Ophytrium significantly atopic and allergic dermatitis in 1-year-old dogs and improved skin health and pruritus in these atopic dogs, has shown significant improvement in skin condition in resulting in high satisfaction levels for veterinarians and controlled field studies. To objectively compare efficacy owners. of oclacitinib to prednisolone, a study was performed Source of funding: Ceva Sante Animale, Libourne, in a laboratory model of allergic dermatitis. House dust France. mite (HDM)-sensitive beagles were exposed to topical Conflict of interest: All authors received reimbursements, HDM allergen (15 mg/mL), and erythema, papules and fees, funding or salary from Ceva Sante Animale. excoriations were scored with a 0 to 5 scale (0, normal; 5, severe). Based on total lesion scores (erythema+papules+excoriations), 21 animals were randomized into treatment groups (n = 7 per treatment): FC-30 placebo (0.0 mg/kg per os twice daily), oclacitinib (Apoquel, Zoetis; Kalamazoo, MI, USA; 0.4–0.6 mg/kg Evaluation of a new hydrolysed fish and rice p.o. twice daily) or prednisolone (0.5 mg/kg p.o. once starch elimination diet in the diagnosis of daily for five days, then tapered every other day). Sub- adverse food reactions in cats sequently, animals were exposed to HDM, then treated < † for 12 days. Oclacitinib significantly (P 0.05) C. NOLI* and G. BELTRANDO decreased all measures compared to placebo-treated *Servizi Dermatologici Veterinari, Peveragno (CN), controls. Area under the curve (mean Æ SEM) for ery- † Italy; Universita degli SAtudi di Torino, Grugliasco thema, papules, excoriations and total lesion scores in (TO), Italy placebo-treated animals were 278.5 (Æ 22.8), 141.9 Diagnosis of food allergy (FA) is based on a two- (Æ 30.5), 111.6 (Æ 23.9), and 532.0 (Æ 76.6), respec- month elimination diet (ED) and confirmed by positive tively, compared to 207.5 (Æ 21.2), 47.2 (Æ 5.9), 46.8 reactions to challenge with the old food. Hydrolysed (Æ 13.7) and 301.6 (Æ 35.8) respectively, in oclacitinib- diets are commonly used for this purpose; unfortu- treated animals. Prednisolone did not have significant nately, they may contain partially hydrolysed allergenic effects on lesion scores. Pruritic behaviour also was ingredients which may bias the results. The selection of quantified, and both oclacitinib and prednisolone the right ED is mandatory for a correct diagnosis. The showed significant (P < 0.05) reductions in pruritic aim of this study was to evaluate the suitability of events compared to placebo. These data support previ- hydrolysed fish and rice starch diet (Vet-Life Feline ous findings with oclacitinib and suggest that it has Ultra-Hypo, Farmina Pet Food; Nola, Italy) in the strong anti-inflammatory properties along with antipru- diagnosis of FA in cats. Cats with suspected FA were ritic effects. enrolled in the study. For each cat, pruritus was Source of Funding: Zoetis, Kalamazoo, MI, USA. assessed by a Visual Analog Scale and lesions by Scor- Conflict of Interest: AJG and JTR are employees of ing Feline Allergic Dermatitis (SCORFAD) on day (D) Zoetis. 0 and D56 of ED. Cats showing ≥50% pruritus and/or SCORFAD improvement were considered FA and challenged with the old food, fish and rice. Cats not responding to the ED were administered a second diet trial (Anallergenic, Royal Canin; Aimargues, France). Twenty-six cats were included, one was excluded for noncompliance and two were lost to follow-up. Of the remaining 23, three interrupted the study for gastrointestinal signs, and 14 (61%) improved and underwent the dietary challenge. Of these, 5/14 (36%) reacted to their old food and were diagnosed with FA; two of these reacted to fish and four to rice. No

FC-32 were within 10 mm of the digital pVAS on a scale of 1–100 mm. Comparing the mean scores obtained on Is a digital pVAS equivalent to a paper PVAS the two modes of administration from the same person to measure itch in dogs suffering from Atopic can be used to assess measurement equivalence. In Dermatitis? 2017, dog owners from 31 different veterinary clinics in Australia with 77 unique dogs diagnosed with atopic A. WRIGHT*, S. SPIEGEL†, K. KAMINSKI †,M. † ‡ dermatitis completed the paper pVAS at the veterinary KARWALA and P. HILL office. Then a code to the digital pVAS app (Itch * Zoetis, Inc, Parsipanny, NJ, USA; † Tivix, Wroclaw, ‡ Tracker) was provided with an initial injection of loki- Poland; Veterinary Dermatology and Immunology, vetmab (Cytopoint, Zoetis Inc.; Parsipanny, NJ, USA) Adelaide, South Australia, Australia and pVAS completed upon download on a mobile An ePRO (electronic patient reported patient outcome) phone. Mean average from the app pVAS was – measure that has been adapted from a paper-based 1.5 mm below paper pVAS mean, much smaller than measure should produce data that are equivalent or the allowable 10 mm. Therefore, the app-based pVAS superior to the data produced from the original paper was shown to be equivalent to the paper-based pVAS version. This prospective study was performed to show and can be used by owners to track pruritus at home. measurement equivalence of a new digital pruritus This may assist veterinarians and dog owners track Visual Analog Scale (pVAS) in an app (Itch Tracker) response to pruritus therapy in between visits to the to the widely accepted paper pVAS following recom- veterinary clinic. mendations of the ISPOR ePRO Good Research Prac- Source of funding: Zoetis, Parsipanny, NJ, USA. tices Task Force. Equivalence was determined to be if Conflict of interest: AW is an employee of Zoetis. scores from dog owners completing the PVAS on paper

FC-33 FC-34 Antimicrobial resistance gene and multidrug Survey of dermatophytes in stray dogs and efflux pumps against antiseptics for cats in Puerto Rico confirmed with MALDI- staphylococci isolated from canine superficial TOF pyoderma in Japan A. HERNANDEZ-BURES*, W. A. BIDOT†,M. ‡ ‡§ N. MURAYAMA*, Y. OSHIMA-TERADA*, K. O’DELL , C. W. MADDOX , W. E. SANDER* and MORI*, H. NAKAMINAMI†, T. SAITO† and N. J.B. PIEPER* NOGUCHI† *Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois, Urbana, IL, *Dermatology Services for Dogs and Cats, Tokyo, † † USA; Animal Care Services, University of Florida, Japan; Tokyo University of Pharmacy and Life ‡ Sciences, Tokyo, Japan Gainesville, FL, USA; Veterinary Diagnostic Laboratory, College of Veterinary Medicine, University Meticillin-resistant staphylococci in canine pyoderma is of Illinois, Urbana, IL, USA; §Department of a serious problem. The aim of this study was to access Pathobiology, College of Veterinary Medicine, mecA, antimicrobial resistance genes and multidrug University of Illinois, Urbana, IL, USA efflux pump genes of staphylococci from canine superficial pyoderma in Japan. One hundred and thirty-six Dermatophytosis is a contagious fungal skin condition dogs were evaluated from May 2018 to April 2019. Sta- of high zoonotic importance and this raises public phylococci mecA gene, antimicrobial resistance genes health concern in working environments with large (including ermA, ermB, ermC, msr, inuA, mphC, aacA/ influxes of stray animals. After hurricanes impacted aphD, tetM, tetK and fosB) and multidrug efflux pump Puerto Rico in 2017, there has been an increased num- genes including qacA/B and smr were identified by ber of stray animals due to owner abandonment, which Staphylococcus pseudinter- has led to veterinary groups visiting the island to per- PCR. Of these, 72% were – medius, 25% S. schleiferi ssp. coagulans and 3% form trap spay/neuter events. The purpose of this unidentified staphylococci, respectively, and 47% of all study was to establish the prevalence of stray animals staphylococci had mecA positive. All staphylococci with dermatophytes and the prevalence of asymp- were positive for ermA, ermB, ermC, msr, inuA, mphC, tomatic carriers. Hair samples were collected from 55 stray dogs and 44 stray cats (total 99) during a trap– aacA/aphD, tetM, tetK and fosB in 0%, 46.3%, 0.7%, – 0%, 0%, 0%, 50.7%, 46.3%, 1.5% and 70.0%, respec- neuter release event using the Mackenzie brush tech- tively. MecA-negative staphylococci were positive for nique and cultured on Derm-duet plates. Direct micro- ermA, ermB, ermC, msr, lnuA, mphC, aacA/aphD, tetM, scopy and examination of the isolates were performed. tetK and fosB in 0%, 22.2%, 0%, 0%, 0%, 0%, In addition, isolates were evaluated by matrix-assisted 27.8%, 33.3, 2.8% and 62.5%, respectively. MecA-posi- laser desorption/ionization-time-of-flight (MALDI- tive staphylococci was positive for ermA, ermB, ermC, TOF). This project was approved by the Institutional msr, lnuA, mphC, aacA/aphD, tetM, tetK and fosB in Animal Care and Use Committee. A total of 19/99 ani- 0%, 74.6%, 1.6%, 0%, 0%, 0%, 76.6%, 60.9%, 0% mals (19%) were positive for Microsporum canis (18/19) and 78.1%, respectively. Multidrug efflux pump genes and Trichophyton spp. (1/19). Prevalence rates of including qacA/B and smr were 0.7% and 2.2% (respec- 29.5% in feline patients and 10.9% in canine patients were observed, approaching statistical significance tively) in all isolates. MecA-positive staphylococci were = = positive for qacA/B and smr in 1.6% and 1.6%, respec- (OR 2.71 feline, P 0.096). Positive cultures were tively. MecA-negative staphylococci were positive for observed in 9.8% of adult patients and 34.2% of juvenile patients, which was statistically significant smr in 2.8%. This study indicated that mecA-positive = = staphylococci had many antimicrobial resistance genes, (OR 3.48 juvenile, P 0.02). Asymptomatic patients and a few staphylococci had multidrug efflux pump were positive in 36% of the population while animals genes. with clinical lesions were positive in 13.5%, which was approaching significance (OR = 0.38 clinical lesions, Source of funding: Self-funded. = Conflict of interest: None declared. P 0.094). This study demonstrates the importance of establishing safety protocols among animals admitted to trap–spay/neuter events for stray animals, regardless of lesions. Source of Funding: Self-funded. Conflict of interest: None declared.

FC-35 the state of Rio de Janeiro, Brazil, affecting mainly cats. Various antifungals are available to treat this dis- Determination of resistance to antimicrobial ease with itraconazole currently being the drug of agents and resistance to meticillin in choice. However, the number of cases refractory to Staphylococcus pseudintermedius isolates from treatment has increased, possibly because of drug resis- cutaneous infections in dogs tance or the quality of the compounded itraconazole often used to treat the disease. The study objective was M. SILVA*, O. VIEIRA-DA-MOTTA†,A. ‡ ‡ to compare different protocols using itraconazole to FIGUEIREDO , F. FERREIRA and M. SILVA- treat feline sporotrichosis. A total of 107 cats from the CARVALHO‡ city of Vassouras diagnosed with sporotrichosis *Estacio de Sa University (UNESA), Rio de Janeiro, between 2016 and 2018 were included based on positive Brazil; †State University of Norte Fluminense, Rio de ‡ cytological findings and fungal culture. Disease extent Janeiro, Brazil; Federal University of Rio de Janeiro, and severity varied from a single skin lesion to multiple Rio de Janeiro, Brazil lesions affecting the skin and mucosa and/or respira- Resistance to antimicrobial agents in Staphylococcus tory tract. The cats were randomly assigned to receive spp. was investigated in 124 study dogs. Over 40% the following treatment protocols: (i) generic itracona- resistance rates were observed against erythromycin zole (Prati Donaduzzi; Parana, Brazil) at 100 mg/cat/ (48.3%), penicillin (45.9%), clindamycin (42.7%) and day per os; (ii) compounded itraconazole at 100 mg/ tetracycline (37.9%). Between 20 and 40% resistance cat/day p.o.; and (iii) generic or compounded itracona- were observed for trimethoprim-sulfamethoxazole and zole at 100 mg/cat/day p.o. combined with 5 mg/kg/ tetracycline. The lowest resistance rates (<15%) were day p.o. of compounded potassium iodide. Response to observed for oxacillin, gentamicin, ciprofloxacin, chlo- treatment was evaluated after 150 days of therapy. ramphenicol, rifampicin and teicoplanin. Likewise, it There were no statistical differences (chi-square test, was found among samples of S. pseudintermedius that P < 0.05) in the number of cats that achieved cure, died resistance to penicillin (52.3%) and erythromycin spontaneously or were euthanized, between the treat- (51.1%) were greatest, followed by clindamycin resis- ment protocols. Based on the study results, brand name tance (40.6%). The lowest resistance (10%) also was or compounded itraconazole is an effective treatment observed for gentamicin, chloramphenicol, rifampicin for feline sporotrichosis if administered for a long per- and teicoplanin. Regarding the detection of meticillin- iod of time. resistant (MR) Staphylococcus strains, 17 of 124 sam- Source of funding: Coordenacßao~ de Aperfeicßoamento de ples were identified as resistant to meticillin by pheno- Pessoal de Nıvel Superior (CAPES), Brazil. typic methods: six among dogs with pyoderma, four Conflict of interest: None declared. among animals with otitis, six with cutaneous tumour and one with surgical site infection. In addition, PCR was used on all samples to detect the mecA gene and confirm meticillin resistance. Of the 17 isolated MR FC-37 samples, the majority (n = 13) belonged to the genomic islands and were identified by PCR as S. pseudinter- Feline sporotrichosis outbreak in Curitiba, medius. The remaining samples were identified by PCR Brazil = † as S. aureus (n 4). Defining multiresistance as resis- D. KUNG*, F. MONTI , L. BALLARDIN*, S. tance to three or more classes of antimicrobials, among RIBEIRO*, L. CONDAS‡, M. CASTRO§,G. the isolated MR S. aureus (MRSA) samples, 75% (3/4) FERREIRA*, S. AMADEI§ and M. FARIAS* showed multidrug resistance. With regard to MR *School of Life Sciences, Pontifıcia Universidade S. pseudintermedius (MRSP) samples, a high rate of Catolica do Parana, Curitiba, PR, Brazil; †Universidade multidrug resistance also was observed (92%, 12/13). Tuiuti, Parana, Brazil; ‡Self-employed veterinarian, Source of Funding: None declared. Parana, Brazil; §Biotechnology, School of Life Sciences, Conflict of interest: None declared. Pontifıcia Universidade Catolica do Parana, Parana, Brazil The present study evaluates the outbreak of 295 cases FC-36 of feline sporotrichosis caused by Sporothrix brasiliensis in Curitiba, southern Brazil, from May 2016 to June The comparative efficacy of different 2019. Cytopathological or histopathological and myco- itraconazole treatment protocols for the logical tests were performed for the final diagnosis. treatment of feline sporotrichosis in Rio de Molecular identification of isolates was performed Janeiro, Brazil using the calmodulin gene species-specific region. The demographic variables analysed included breed, age, † ‡ § F. CARVALHO*, , M. MACHADO , S. KOCH , sex, reproductive status, outdoors access, soil/decay ‡ ‡ R. RAMADINHA and J. FERNANDES vegetation contact, cohabitation and contact with out- *College of Veterinary Medicine, Centro Universitario de doors animals. Clinical data included disease duration † Valencßa, Valencßa, Rio de Janeiro, Brazil; College of before diagnosis, lesion classification and topography. Veterinary Medicine, Universidade Iguacßu, Nova Iguacßu, Zoonotic data included type of transmission, topogra- ‡ Rio de Janeiro, Brazil; Department of Veterinary phy and classification of lesions. Demographic data Science, Universidade Federal Rural do Rio de Janeiro, showed that 275 (93%) cats lived in urban area, 283 § Seropedica, Rio de Janeiro, Brazil; College of Veterinary (96%) were mixed breed of adult age (average 44- Medicine, University of Minnesota, MN, USA month-old), 203 (69%) were males, 160 (58%) were Sporotrichosis caused by an agent of the Sporothrix intact, 223 (77%) had outdoor access and 231 (84%) schenckii complex, is an important endemic zoonosis in had contact with soil/decay vegetation. The disease

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 42 Abstracts duration before diagnosis for 275 cats was over 14 FC-39 weeks. The head and forelimbs were affected more frequently. Sixty-nine (25.3%) of 272 owners developed Evaluation of the response to therapeutic sporotrichosis from contact with exudate and scratches protocols in feline sporotrichosis or bites from affected cats. Lesions developed mainly F. MONTI, L. BALLARDIN, S. RIBEIRO, D. on their upper limbs and typically in a localized form. KUNG, V. GMYTERCO, C. PIMPAO~ and M. In conclusion, intact male and adult cats represent the FARIAS population most exposed to S. brasiliensis in Curitiba. Department of Veterinary Medicine, School of Life These animals develop disseminated cutaneous lesions Sciences, Pontifical Catholic University of Parana, and are the most important host and propagator of Parana, Brazil sporotrichosis in urban environments in this state. Source of funding: Coordenacßao~ de Aperfeicßoamento de Sporotrichosis is an epidemic mycotic infection in Bra- Pessoal de Nıvel Superior (CAPES). zil that affects primarily cats living in urban areas. The Conflict of interest: None declared. aim of this longitudinal study was to evaluate the efficacy of different therapeutic protocols in cats with sporotrichosis caused by Sporothrix brasiliensis. Fifty- nine cats diagnosed with sporotrichosis were enrolled in FC-38 the study. Thirty-four were treated with generic itraconazole (ITZ) monotherapy at the dose of 10 20 mg/ Antifungal susceptibility of Sporothrix kg/day per os in patients ≤3.0 kg or 100 mg/cat/day brasiliensis isolated from feline sporotrichosis p.o. in patients >3.0 kg. Twenty-five cats were treated with ITZ using the same treatment protocol combined D. C. KUNG*, F. MONTI*, L. BALLARDIN*, L. † ‡ ‡ with compounded potassium iodide (KI) at the dose of CONDAS , L. BORBA-SANTOS , E. MENEZES ,S. ‡ 5 mg/kg/day p.o. Among the 34 cats treated solely with ROZENTAL and M. FARIAS* ITZ, 11 (32%) were cured, one (3%) was euthanized *Postgraduate Program in Animal Science, School of and the other 22 (65%) required the addition of KI to Life Sciences, Pontifıcia Universidade Catolica do † their therapeutic protocol. Of the 47 cats treated with Parana, Parana, Brazil; Freelance veterinarian, Parana, ‡ the combination of ITZ and KI, 24 (51%) were cured, Brazil; Laboratory of Fungal Cell Biology, Institute of seven (15%) died and 16 (34%) required the addition Biophysics Carlos Chagas Filho, Federal University of of intralesional amphotericin B deoxycholate (AMB). Rio de Janeiro, Rio de Janeiro, Brazil When AMB was added to the treatment regimen, nine This study aimed to evaluate the susceptibility of Spor- (56%) of the 16 cats recovered from the disease. The othrix brasiliensis, isolated from cats with sporotri- overall cure rate was 74.5% (44/59 cats). In conclusion, chosis, to itraconazole (ITZ) and relate it to disease the combination of ITZ and KI therapy was necessary severity and outcome. The diagnosis was confirmed in most cats to achieve disease cure. In some cases, based on cytopathological or histopathological and however, the addition of a third drug (intralesional mycological test results. Molecular identification of iso- AMB) to the treatment regimen was needed to improve lates was performed using the calmodulin gene species- success rate. specific region. Susceptibility tests were preceded by fil- Source of funding: Coordination for the improvement of amentous to yeast conversion according to Clinical and higher education personnel (CAPES), Brazil. Laboratory Standards Institute (CLSI) protocols. Fungi Conflict of interest: None declared. are here designated as wild-type (WT) for those without intrinsic or acquired resistance and nonWT (NWT) for those harbouring intrinsic or acquired resistance mechanisms. Of the 67 cats included in the study, all were FC-40 infected with S. brasiliensis. Itraconazole susceptibility was performed in 37(54.4%) isolates that converted Performance of a pad containing Ophytrium from filamentous to yeast. Complete clinical data were and chlorhexidine digluconate 3% in dogs with available for only 17(46%) of these isolates. Twelve local bacterial and/or Malassezia overgrowth (70.5%) of these were WT and five (29.5%) NWT. Ten † (83.3%) cats with WT isolates were cured; however, M. GATELLET*, R. KESTEMAN*, B. BAULEZ ,F. PRADIES†, P. M. CADOT‡, H. DROPSY§,P. eight (80%) were cured with ITZ combined to potas- ¶ †† sium iodide (KI). Two (16.7%) cats were euthanized. FIORA , J. L. MATHET , E. OLLIVIER*, C. Three cats with sporotrichosis caused by the NWT iso- BILLY* and X. DE-JAEGER* † late were cured with ITZ and KI combined, and two *Ceva Sante Animale, Libourne, France; Clinique died from the disease. Thirteen (76.5%) of the 17 cases veterinaire VPlus, Saint Aubin de Blaye, France; ‡ § had the disseminated cutaneous form associated with Clinique veterinaire SeineVet, Rouen & Boos, France; respiratory clinical signs. In conclusion, most cats Ecole nationale veterinaire d’Alfort, Maisons-Alfort, ¶ infected with ITZ-susceptible S. brasiliensis strains France; Clinique veterinaire Saint-Jacques, Agen, †† required combination therapy (i.e. ITZ and KI) to France; Clinique veterinaire des Glycines, Orleans, achieve disease cure. Unfavourable prognosis was most France often associated with ITZ-resistant S. brasiliensis Local disturbances of the microbiota are common in strains. dogs with underlying skin conditions. Antiseptic topical Source of funding: Coordenacßao~ de Aperfeicßoamento de products are indicated to control such superficial disor- Pessoal de Nıvel Superior – (CAPES). ders. The objective of this study was to evaluate the Conflict of interest: None declared. performance of a daily application of pads containing Ophytrium and chlorhexidine digluconate 3% (DOUXO S3 PYO Pads, Ceva Sante Animale;

Libourne, France) in dogs with focal bacterial and/or SBF does not always represent the primary pathogen Malassezia overgrowth. Eighteen dogs with focal skin causing the disease. Bacterial culture and antibiotic sus- dysbiosis were enrolled in a prospective, multicentric, ceptibility testing from multiple lesions are recom- open study. Dogs received daily pad applications for 14 mended when S. schleiferi is isolated from a pustule in days. Bacterial and/or Malassezia counts per micro- canine SBF. scopic field and a global score of the most affected area Source of funding: Self-funded. (0–17 scale based on extension, severity, bacterial and Conflict of interest: None declared. Malassezia cytological scores) were assessed by a veterinarian and pruritus by the owner (pruritus Visual Ana- log Scale, pVAS) on day (D)0, D7, D14. Owner and veterinarian evaluations of performance and satisfac- FC-42 tion were recorded. Eleven dogs had primarily cocci overgrowth and seven mostly Malassezia. Mean bacte- Evaluation of the effects of chlorhexidine rial and Malassezia counts decreased after 14 days (6.9 digluconate with and without canine b-defensin to 1.1; 7.6 to 1.5 respectively); 88.9% of dogs achieved 103 or canine cathelicidin against multidrug- a ≥ 70% microbial decrease. Mean global score of the resistant clinical isolates of Staphylococcus most affected area and pVAS significantly improved at pseudintermedius D14, respectively, from 8.6 to 2.6 and 4.5 to 1.2 † < L. KHER*, D. SANTORO*, V. CHALA and (P 0.05 each; mean improvements of 70.4% and † 71.4%, respectively). Global veterinary assessment of C. NAVARRO the protocol was satisfactory, good or excellent in *Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, 88.9% of cases. Most owners (94.4%) considered the † protocol efficacious. Using a pad containing Ophytrium Gainesville, FL, USA; Virbac SA Laboratories, Carros, and chlorhexidine digluconate 3% daily for 14 days France improved the skin condition and pruritus of dogs with Host defence peptides (HDPs) such as b-defensin (BD) local dysbiosis, resulting in high satisfaction levels for and cathelicidin (Cath) play a fundamental role in the both veterinarians and dog owners. epithelial natural defence against micro-organisms. Source of funding: Ceva Sante Animale, Libourne, Because of their mechanism of action, HDPs have France. shown very low bacterial, fungal and viral resistance. Conflict of interest: All the authors received reimburse- Our objective was to determine if their presence might ments, fees, funding or salary from Ceva Sante Animale. facilitate the antimicrobial activity of chlorhexidine against multidrug-resistant Staphylococcus pseudintermedius (MRSP) in in vitro conditions. Ten MRSP clinical isolates were used for this study. Two-fold serial FC-41 dilutions of 2% chlorhexidine solution (1:32,000, 1:64,000, 1:128,000 and 1:256,000) alone or in combina- Significance of isolation of Staphylococcus tion with synthetic canine HDPs (cBD103 and cCath) schleiferi in canine superficial bacterial were prepared. A concentration of 1 µg/mL of syn- folliculitis thetic HDP was selected based on previous studies. T. OSUMI*,†, Y. SHINOMIYA*, I. IMANISHI*, Y. Friedman’s test and a Dunn’s multiple comparison test SHIMAZAKI†, K. IDE*,†, K. IYORI‡ and K. were used for statistical analysis. Minimal inhibitory † NISHIFUJI*, concentration 90 (MIC90) after 24 h for chlorhexidine † µ *Laboratory of Veterinary Internal Medicine; Animal [1:64,000 dilution (0.3125 g/mL)] alone or in combina- Medical Centre, Tokyo University of Agriculture and tion with HDPs were obtained. Minimal bactericidal Technology, Tokyo, Japan ‡Vet Dem Tokyo, Kanagawa, concentration was achieved only for chlorhexidine com- µ Japan bined with cCath at 1:32,000 (0.625 g/mL) and 1:64,000 (0.3125 µg/mL) dilution after 30 and 60 min Canine superficial bacterial folliculitis (SBF) is a bacte- exposure, respectively. A significant reduction rial skin disease characterized by follicular papules and (P < 0.05) of colony forming units was seen for the pustules. Although Staphylococcus pseudintermedius is same combination compared to chlorhexidine alone for the most frequently isolated bacterial species, most concentrations and time points tested (5, 15, 30 S. schleiferi is occasionally isolated from skin lesions of and 60 min). A 2-log10 reduction (synergistic effect) canine SBF. The pathogenic involvement of S. schleiferi was achieved for the highest concentrations (1:32,000 in canine SBF has not been sufficiently investigated. In and 1:64,000) after 30 and 60 min, respectively. These this study, we investigated whether S. schleiferi can be preliminary data suggest the potentiation of chlorhexi- solely isolated from pustules in canine SBF. Initially, dine on MRSP when associated to cCath. Potential seven staphylococcal isolates were obtained from seven mechanics could involve cell membrane disruptive pustules in a dog diagnosed with SBF and subjected to properties of HDPs. multiplex PCR to identify the staphylococcal species. Source of funding: Virbac Corporation, Carros, France. One isolate was S. schleiferi, whereas the remaining six Conflict of interest: DS received reimbursements, fees, were S. pseudintermedius. A total of 36 staphylococcal funding or salary from Virbac. isolates were obtained from four pustules in the same dog three months after the first study and subjected to the same experiment. Four of 12 isolates obtained from a single pustule were S. schleiferi, while the remaining 32 obtained from all four pustules were S. pseudintermedius. These results imply that identification of S. schleiferi obtained from a single pustule of canine

FC-43 diffusion inhibitory zone of 15–20 mm. This study aimed to determine the optimum breakpoint of oxacil- Efficacy of proprietary formulations containing lin disk diffusion inhibitory zone specific to S. schleiferi essential oils and plant extracts compared to subsp. coagulans (SSC) and to determine whether the chlorhexidine plus miconazole shampoo in value represents a clinical breakpoint for cefalexin canine superficial pyoderma against SSC in canine pyoderma. Receiver operating characteristic curve analysis revealed that the best C. ROMERO*, G. SHEINBERG†,A.M. § breakpoint of disk diffusion inhibitory testing against CORDERO and R. HEREDIA* SSC was achieved when it was approximately 22 mm *DERMAVET, Iztapalapa, Mexico; †Centro Veterinario = = § (Youden’s index 0.89, n 93). Next, 25 dogs with Mexico, Mexico City, Mexico; VETDERM, pyoderma, in which oxacillin-susceptible (≥23 mm, Guadalajara, Mexico n = 9), intermediate (18–22 mm, n = 6) and resistant Superficial pyoderma is a common condition in dogs. (≤17 mm, n = 10) SSC strains were isolated from skin Antibiotic and potentially antiseptic resistance is a lesions, were treated with oral cefalexin for 14 days. growing problem and natural topical formulations may There was a significant difference in the improvement provide an alternative option. Three natural formula- rate of clinical scores between oxacillin-susceptible and tions with essential oils and plant extracts were com- -intermediate groups (Friedman test: P < 0.01), whereas pared to chlorhexidine + miconazole and microsilver there were no significant differences between intermedi- shampoo. Thirty dogs with clinical and cytological evi- ate and resistance groups (P > 0.05). These findings dence of superficial pyoderma were randomized into indicate that the oxacillin disk diffusion breakpoint of three groups: 1, PYOclean shampoo (LDCA; Castres, ≤22 mm is the optimum clinical breakpoint to predict France) twice weekly and daily rinse-free PYOclean the efficacy of cefalexin in dogs with pyoderma caused Mousse (LDCA); 2, PYOclean shampoo twice weekly by SSC. and daily use of PYOclean Spray; and 3, Biohex sham- Source of funding: Self-funded. poo (Vetbiotek; Largo, FL, USA) twice weekly. Cyto- Conflict of interest: None declared. logical and lesion scores, and pruritus Visual Analog Score (pVAS) were performed on day (D)1, D7 and D14. Bacterial numbers decreased by 100% on D7 and D14 in groups 1 and 2, and 88.89% on D7 and 100% FC-45 on D14 in Group 3. Yeast numbers decreased on D7 A case of multiresistant staphylococcal and D14 by 45.46% and 81.82% (respectively) in Group 1, 76% and 92% in Group 2, and 50% and infection of the ear successfully treated with 90% in Group 3. Lesion scores decreased on D7 and diode laser and photodynamic treatment with D14 by 70.27% and 90.61% in Group 1, 52.49% and infracyanine green 87.95% in Group 2, and 33.55% and 62.94% in Group A. VERCELLI, E. GUIDI, A. CORONA and L. 3. The pVAS decreased on D7 and D14 by 74.65% and CORNEGLIANI 95.78% in Group 1, 56.63% and 86.75% in group 2, Clinica Veterinaria Citta Di Torino, Torino, Italy and 71.09% and 69.88% in Group 3. We conclude that these plant-based products can be an effective alterna- Photodynamic treatment with infracyanine green and tive to chlorhexidine for topical treatment of superficial diode laser is used in human and veterinary dentistry pyoderma in dogs. for the treatment of bacterial infections. A 10-year-old Source of funding: Dermoscent Laboratoire (Laboratoire male West Highland white terrier with chronic unilat- de Dermo-cosmetique Animal), Castres, France. eral otitis and a history of unsuccessful systemic and Conflict of interest: None declared. topical antibiotic treatments was referred. On physical examination, several skin fistulas were present adjacent to the ear base; the ear canal was completely stenotic and otoscopy could not be performed. Computer FC-44 tomography revealed calcification of the ear canal with ruptured tympanic membrane, abscessation, osteomyeli- Re-evaluation of oxacillin disk diffusion tis of the bulla and temporal condylus. Based on bacte- breakpoints for detection of mecA-mediated rial culture and susceptibility Staphylococcus oxacillin resistance in Staphylococcus schleiferi pseudintermedius sensitive to pradofloxacin was iso- isolated from dogs lated. Systemic treatment with oral pradofloxacin 3 mg/ kg once daily improved the infection and the dog had Y. SHIMAZAKI*, K. SHIMOIKE†, K. NISHIFUJI*,‡ † total ear canal ablation and bulla osteotomy per- and K. IYORI formed. After one month, despite antibiotic treatment *Animal Medical Centre, Tokyo University of new fistulas developed at the same area. Bacterial cul- Agriculture and Technology, Tokyo, Japan; †Vet Derm ‡ ture revealed a multiresistant S. pseudintermedius sus- Tokyo, Kanagawa, Japan; Laboratory of Veterinary ceptible to rifampicin. Under general anaesthesia, the Internal Medicine, Tokyo University of Agriculture and area was cleaned with 0.5% saline solution and 1 mL Technology, Tokyo, Japan of infracyanine green (Emundo, A.R.C. GmbH; Nurn- The recent guidelines for antimicrobial susceptibility berg, Germany) was injected into the fistulas. A treat- testing for oxacillin reveal that the breakpoints of disk ment with diode laser (A.R.C. Laser 810 nm) of four diffusion inhibitory zone (≤17 mm) against Staphylo- cycles 500 MW for 30 s per centimetre of affected area coccus pseudintermedius also could be applied to meti- was performed. No antibiotic was administered and cillin-resistant S. schleiferi (MRSS). However, the one week later, 50% of the fistulas were closed with previous study used an insufficient number of significant improvement of the patient’s clinical condi- S. schleiferi isolates that showed an oxacillin disk tion. Cytological results and bacterial culture were

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 Abstracts 45 negative 72 h post-treatment. Total recovery occurred objectives of this study were to find the most common after two weeks. Photodynamic treatment is a promis- staphylococci isolated from cats and dogs as well as the ing antibacterial technique in case of localized refrac- resistance patterns for these bacteria. All staphylococcal tory bacterial skin infections. isolates submitted during the study period were Source of funding: Self-funded. included. Cultured bacteria were analysed by matrix- Conflict of interest: None declared. assisted laser desorption-ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and identified by BIO- TYPER (v2.0) software (Biomerieux; Marcy-l’Etoile, France). Sensitivity was determined by disk diffusion FC-46 following Clinical and Laboratory Standards Institute (CLSI) guidelines M100-S26, M100-S27 and VET01S3 Adhesive tape impression cytology in recommendations for standard antibiotics. In total, 17 dermatophytosis and pemphigus foliaceus species of staphylococci were identified. Staphylococcus L. J. VOGELNEST*,†, C. B. LUDWIG* and P. A. felis was most frequently isolated from cats at 54% fol- RAVENS* lowed by S. pseudintermedius at 16%. Staphylococcus *Small Animal Specialist Hospital, North Ryde, NSW pseudintermedius accounted for 64% and S. schleiferi Australia; †School of Veterinary Science, University of 30% of the isolates from dogs. For all species of Sydney, NSW Australia staphylococci, resistance rates were as follows: penicillin 61%, azithromycin 41%, doxycycline 37%, clindamycin Adhesive tape impression (ATI) cytological evaluation 28%, enrofloxacin 18%, marbofloxacin 18% and meti- has recognised value in veterinary dermatology for cillin 5%. No isolates tested were resistant to baci- diagnosis and monitoring response to therapy of bacte- tracin. Amikacin resistance was 0.5% and rifampicin rial and yeast overgrowth, and some superficial para- resistance was 1%. For S. felis isolated from cats, resis- sitic infestations. However, suitability of ATI for tance rates were as follows: penicillin 50%, azithromy- diagnosis and monitoring response to therapy of der- cin 29%, clindamycin 16%, doxycycline 7%, meticillin matophytosis or pemphigus foliaceus (PF) is poorly 0%, enrofloxacin 0% and marbofloxacin 0%. For evaluated. This retrospective study evaluated ATI find- S. pseudintermedius isolated from dogs, resistance rates ings in dogs and cats with confirmed dermatophytosis were as follows: penicillin 77%, doxycycline 66%, azi- or PF. Patient records were reviewed for 10 dogs and thromycin 61%, clindamycin 49%, enrofloxacin 28%, 10 cats with dermatophytosis confirmed by culture, marbofloxacin 27% and meticillin 4%. Overall, histopathological evaluation and/or complete response S. pseudintermedius was the most common staphylococ- to antifungal therapy. Where culture was performed, cus isolated with low meticillin resistance, and high Microsporum canis or M. gypseum were isolated. Fun- doxycycline and azithromycin resistance. gal elements were reported on ATI in 10/10 cats and 8/ Source of funding: Self-funded. 10 dogs. Arthrospores were recorded as moderate to Conflict of interest: None declared. numerous in 10 cats, located within and clustered around hair shaft fragments and keratinocytes. Fungal hyphae were recorded as sparse within hair shafts of some cats. Fungal hyphae were reported in 8/10 dogs, and typically recorded as focal or sparse and located FC-48 on keratinocytes associated with clusters of neutrophils Expression of antimicrobial peptides and (most commonly) or within hair shafts. Arthrospores cytokines following bacterial stimulation in an were not reported in any dog. Patient records also were ex vivo skin explant model evaluated for 10 dogs and 10 cats with PF confirmed † ‡ § by histopathological evaluation. Acantholytic ker- J. E. HYUN*, Y. B. KIM , and C. Y. HWANG atinocytes were reported associated with neutrophil *Department of Veterinary Internal Medicine, Konkuk clusters in all 20 cases, typically recorded as few-to- Veterinary Medical Teaching Hospital, Konkuk † moderate in cats and moderate-to-numerous in dogs. University, Seoul, Republic of Korea; BON Animal Adhesive tape impressions appear valuable for the diag- Medical Center, Gyeonggi-do, Republic of Korea; ‡ nosis and potential monitoring of dermatophytosis and Department of Veterinary Surgery, College of PF in dogs and cats. Veterinary Medicine, Konkuk University, Seoul, § Source of funding: Self-funded. Republic of Korea; Laboratory of Veterinary Conflict of interest: None declared. Dermatology, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea The skin protects the body by mediating various FC-47 immune responses against exogenous substances includ- Staphylococcal species isolated and ing bacteria, fungi and viruses. In addition, it has a predominant role as a physical barrier. Despite the antimicrobial resistance from canine and feline significant protection offered via various mechanisms, bacterial cultures in Hong Kong bacterial infection of the skin is one of the most com- R. WESTERMEYER mon skin diseases in veterinary medicine. Here, we Animal Allergy Specialists, Kaneohe, Hawaii, USA; and examined the structural and immunological changes in Asia Veterinary Diagnostics, Hong Kong, Hong Kong the skin during infections with Pseudomonas aeruginosa and Staphylococcus pseudintermedius using skin explants This bacterial surveillance study was performed to char- from four healthy beagles. Skin structure was generally acterize all staphylococcal isolates cultured from well preserved in uninfected controls, yet defects in the September 2016 to March 2017 in Hong Kong. The skin structure, including injury of keratinocytes and

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 46 Abstracts dermo-epidermal junction disruption, were identified clinical resistance to all B-lactam antibiotics. The inci- when skin explants were exposed to P. aeruginosa and dence of MRS in cases of canine pyoderma requiring S. pseudintermedius. On exposure to P. aeruginosa, systemic antibiotic therapy in a referral dermatology marked linear cleft formation and acantholysis along practice in Melbourne, Australia, was examined the basal layer was noted after 24 h of culture. In addi- prospectively from September to December in 2014, tion to the skin structure defects, mRNA expression 2016 and 2019. Cases were classified as responders to levels of the antimicrobial peptides (AMP) cBD103 and empirical therapy (RTET), lost to follow-up (LTFU), S100A8 were decreased, which was confirmed by culture-confirmed meticillin-susceptible Staphylococcus immunohistochemical staining. Conversely, mRNA sp. (MSS) or culture-confirmed MRS. Both confirmed expression levels of IL-6, IL-8 and TNF-a were incidence (i.e. based on culture and susceptibility) as increased in S. pseudintermedius-exposed skin explants. well as presumed incidence (i.e. included all LTFU Taken together, these results suggest that the ex vivo cases on the assumption they were MRS and the RTET canine skin model used in this study is a suitable cases assuming they may have been successfully treated research tool for investigating bacterial skin infections MRS cases) were calculated. Trends in antibiotic sus- in dogs. ceptibilities, as well as the incidence of non-S. pseudin- Source of funding: Self-funded. termedius MRS also were examined. Significance was Conflict of interest: None declared. assessed using Fisher’s exact test. Ninety-four, 104 and 123 dogs entered the study in 2014, 2016 and 2019, respectively. The incidence of confirmed (14.4–24.3%) and presumed (38.3–42.7%) MRS pyoderma cases was FC-49 not significantly different in any of the three years. An increase in non-S. pseudintermedius MRS from 2014 Change in incidence and antibiotic (0.0%) to 2019 (16.7%) also was not significant susceptibility of meticillin-resistant (P = 0.1435). However, the number of extensively resis- Staphylococcus from canine pyoderma in tant MRS isolates was significantly greater in 2019 referral practice in Melbourne, Australia from (26.7%) than 2014 (0.0%; P = 0.038), largely due to a 2014 to 2019 reduction in susceptibility to trimethoprim-sulfonamide (P = 0.0106; 2014–2019), minocycline (P = 0.0187; D. ROBSON 2016–2019) and chloramphenicol (P = 0.0283; 2016– Animal Skin and Ear Specialists, Melbourne Veterinary 2019). Canine pyoderma complicated by MRS contin- Specialist Centre, Melbourne, Victoria, Australia ues to be a challenge in clinical practice in Melbourne. Staphylococcus pseudintermedius is the most common Source of funding: Self-funded. cause of canine pyoderma. Meticillin-resistant Staphylo- Conflict of interest: None declared. coccus (MRS) contains the mecA gene and causes

FC-50 the groups. From the 2,069 positive samples, 49.73% (1,029) were positive to insects, 84.73% (1,753) to Prevalence of horses suspect of atopic mites, 65.80% (1,360) to moulds and 75.80% (1,570) to dermatitis could be higher than that of insect pollens. Only 39 (1.88%) samples were exclusively posi- bite hypersensitivity tive to insects. The percentage of samples positive to C. LORENTE-MENDEZ and R. WAGNER insects was lowest from January to April (average Laboklin GmbH & Co. KG, Bad Kissingen, Germany 28.21%), and highest from May to October (average 50.48%). Positive samples for mites (76.41–98.46%) Insect bite hypersensitivity (IBH) is considered the most and pollens (61.76–86.79%) were >60%, and for common allergic skin disease in horses, whereas the moulds (50–80%) >50% throughout the year. The prevalence of atopic dermatitis (AD) is unknown. This prevalence of positive reactions was higher on a retrospective study aimed to evaluate the prevalence of monthly basis for any of the three groups of aeroaller- positive serum immunoglobulin (Ig)E antibodies against gens than for insects. Sole reactions to insects were four groups of allergens: insects, pollens, mites and rare. The results suggest that the prevalence of horses moulds using ELISA technology. Serum samples from suspected of having AD is higher than that of IBH, 2,412 horses in Europe with a possible allergic disease and serum IgE reactive to both allergen groups is com- were tested. Two thousand and sixty-nine (85.78%) mon. samples yielded positive results for at least one of the Source of funding: Self-funded. four groups of allergens and 674 (32.58%) of these Conflict of interest: All authors work for Laboklin. samples were positive to allergens representing all of

FC-51 Visual Analog Scale was used to score the lesions and pruritus, and microscopic observations made for the Efficacy of sarolaner on African hedgehogs presence of the C. tripilis mite. Both animals were trea- (Atelerix albiventris) infested with Caparinia ted with 2 mg/kg of sarolaner (Simparica, Zoetis; Kala- tripilis mazoo, MI, USA) orally, mixed with wet food. Patients A. RANGEL*, C. ROMERO†, G. SHEINBERG‡,R. were evaluated on pruritus level, skin lesions and pres- HEREDIA† and A. CORDERO§ ence of parasites on day (D)1, D7, D14, D30 and D60. *Hospital Veterinario Grandes Companeros~ de Torreon, The data were analysed with the Wilcoxon signed rank Coahuila, Mexico; †Dermavet, Edo Mex, Mexico; test. The severity of lesions in both hedgehogs showed = ‡Centro Veterinario Mexico, CDMX, Mexico; significant decrease (P 0.03) from D1 to D30, with a §Vetderm, Guadalajara, Mexico score of zero from D30 to D60. The itching decreased significantly from D1 to D15 (P = 0.02) reaching and The most common mite in the African pygmy hedgehog remaining zero at that time. The presence of mites (Atelerix albiventris)isCaparinia tripilis, causing lesions decreased significantly (P = 0.03) in both hedgehogs and itching of the head, pinnae, dorsum and the exter- from D1 to D15 post-treatment, reaching zero and nal auditory canal. The objective of this study was to remaining thus until D60. This study demonstrates mite evaluate the efficacy of sarolaner as a treatment for control for 60 days with only one administration of sar- C. tripilis infestation in African pygmy hedgehogs. Two olaner in hedgehogs infested with C. tripilis. African hedgehogs were recruited, a male and a female, Source of funding: Self-funded. nine months old, weighing 350 g, presenting with gener- Conflict of interest: None declared. alized erythema, scaling, pruritus and loss of spikes. A

FC-52 Role of extracellular vesicles in skin diseases: FC-53 potentiality of new diagnostic tests based on Susceptibility of Australian canine otitis the analysis of circulating extracellular vesicles externa bacterial and fungal isolates to L. BONGIOVANNI*,†, A. ANDRIESSEN*, T. A. P. florfenicol and terbinafine DRIEDONKS*, E. N. M. NOLTE-THOEN* and A. S. DE BURGH*, E. HICKEY† and D. TROTT† ,† DE BRUIN* *Bayer Australia Ltd, Pymble, Australia; †University of *Department of Biomolecular Health Science, Faculty of Adelaide, Adelaide, Australia Veterinary Medicine, Utrecht University, Utrecht, the † Most conventional otitis externa (OE) treatments for Netherlands; Department of Pediatrics, University dogs require single/multiple daily applications and Medical Center Groningen, University of Groningen, cleaning by dog owners, until the condition is resolved. Groningen, the Netherlands. Florfenicol/terbinafine/mometasone (Neptra, Bayer; Analagous to messages sent by cells to communicate Leverkusen, Germany) applied as single treatment elim- with each other, extracellular vesicles (EVs) have inates pet-owner noncompliance and reduces the possi- emerged as important mediators of intercellular com- bility of antimicrobial resistance arising from munication in various physiological and pathological incomplete treatment protocols. An Australian survey processes. In human skin diseases such as cancer, espe- identified the most prevalent canine OE bacterial cially cutaneous melanoma, as well as inflammatory pathogens as Pseudomonas aeruginosa, Staphylococcus diseases (such as acne, Malassezia, Leishmania, atopic pseudintermedius, Proteus sp., beta-haemolytic strepto- dermatitis), EVs have been outlined as important play- cocci (e.g. Streptococcus canis) and Escherichia coli. ers in disease pathogenesis and progression. EVs could Malassezia pachydermatis is the most common fungal potentially be clinically applicable as drug-delivery vec- pathogen. This study was undertaken to determine min- tors, cell-free vaccines and immunomodulators. In diag- imum inhibitory concentrations (MICs) of florfenicol nostics, circulating EVs provide a novel approach to and terbinafine against bacterial and fungal isolates the development of biomarkers, carrying information sourced from Australian clinical cases of canine OE. of the tissue/cell of origin and being enriched with dis- Isolates of P. aeruginosa were excluded. All bacterial ease-relevant molecules. In particular, mRNA mole- isolates showed consistent MIC values when tested cules are packaged into EVs in a specific manner. This against florfenicol, resulting in very narrow MIC is why we think that circulating EVs and their mRNA ranges. S. canis and S. pseudintermedius isolates showed cargo can be exploited as a potential diagnostic test in low MIC values of ≤4 lg/mL. Gram-negative isolates canine patients with different type of diseases, such as exhibited slightly higher MIC values against florfenicol cancer. We applied a protocol to isolate EVs from and MICs were still very low overall. Proteus sp. and canine blood samples based on the use of size exclusion E. coli isolate MIC values ranged from 4 to 16 lg/mL, chromatography (SEC) and analyse their RNA content. whereas M. pachydermatis isolates showed a much First, we collected blood from healthy dogs and canine wider range of MIC values against terbinafine (0.125– cancer patients with different types of skin cancer. EVs 32 lg/mL). However, most of these isolates were sus- were isolated by using qEVoriginal/ 70 nm (Izon) SEC ceptible to the lower concentrations in vitro, demon- columns and confirmed by Western blotting; RNA was strated by the low MIC50 and MIC90 values of 0.25 extracted from SEC fraction pools and analysed by and 1 lg/mL, respectively. Recent isolates from cases quantitative reverse transcription (qRT)- PCR. Protocol of canine OE in Australia are susceptible to florfenicol details on how blood samples were collected and pro- and terbinafine at concentrations achieved during topi- cessed, EVs isolated, and RNA extracted and analysed cal delivery of Neptra, at the recommended dose rate. will be shown and explained, discussing current limita- Source of funding: This study was fully funded by Bayer tions and potential applications in veterinary medicine. Australia Ltd. Source of Funding: European Research Council under the Conflict of interest: SdeB is an employee of Bayer Aus- European Union’s Seventh Framework Programme (FP/ tralia Ltd. 2007-2013)/ERC grant number 337581. Conflict of Interest: None declared.

FC-54 wavelength through the 160 x 60 mm rectangular shape probe. The NB-UVB dose was started from a minimum Single myringotomy in the treatment of middle 250 mJ once or twice weekly and then gradually ear otitis in the cat: a retrospective study increased to maximum if previous irradiation was S. DELEPORTE* and P. PRELAUD† unsuccessful. The skin lesions were nasal erythema and/ *Dermatology Department, LORRAINEVET, Ludres, or depigmentation, nasal erosion and/or ulcer, perianal France; †Dermatology Department, ADVETIA Centre erythema and/or depigmentation, perianal erosion and/ Hospitalier Vet erinaire, Velizy, France or ulcer, and erythroderma, with remission rates of 75.0%, 50.0%, 33.3%, 20.0%, 20.0% and 0%, respec- Otitis media occurs frequently cats yet data on medical tively. Effective doses were 500–850 mJ and most cases treatment are lacking except for polyp-associated otitis required once weekly proactive irradiations. Post-trea- media. We studied retrospectively 24 cases of cats with ted skin biopsy collected from one dog demonstrated otitis media confirmed by MRI or CT without polyp or fewer infiltrations of tumour cells. No severe adverse neoplasia. Most cats (58%) were presented with bilat- reactions appeared except for mild erythema (28.5%) eral involvement. Both parts of the bulla were involved or pruritis (28.5%). This study suggested that NB-UVB in 97% of cases. Clinical signs included head tilt 54% therapy is safe and useful for canine CTCL. (11/24), Horner syndrome 29% (7/24), ataxia or otalgia Source of funding: Self-funded. 9% (2/24), otitis externa 21% (5/24), nystagmus and Conflict of interest: None declared. facial paralysis 4% (1/24). Cytological findings of middle ear effusion were predominantly neutrophilic. Cocci were identified on cytological examination in 20/38 samples and rods in 2/38. Bacterial culture was positive in 50% of samples, with isolation of Pseudomonas sp., FC-56 Pasteurella multocida, Staphylococcus felis, S. schleiferi, Detection and inhibition of IgE for cross- S. pseudintermedius and Serratia marcesens. All cats reactive carbohydrate determinants evident in were treated using single myringotomy with soft bulla an ELISA for detection of allergen-specific flushing (0.5–2 mL saline) associated with systemic corticosteroid (prednisolone 1 mg/kg/day, seven–10 days) IgE in the serum of dogs and cats and systemic antibiotics according to susceptibility test- K. LEE*, B. MCKINNEY*, K. BLANKENSHIP*, K. ing (one month). Seventeen cats (71%) were clinically ENCK* and D. MORRIS† cured 60 to 240 days after treatment. One cat was *Stallergenes Greer, Lenoir, NC, USA; †School of healthy with a mild head tilt. One cat was euthanised Veterinary Medicine, University of Pennsylvania, because of failure to control Pseudomonas infection. Philadelphia, PA, USA Another one died because of complications of anaesthe- It has recently been demonstrated that immunoglobulin sia. The four remaining cats were cured after ventral (Ig)E specific for cross-reactive carbohydrate determi- bulla osteotomy. This simple technique with only one nants (CCD) is present in the serum of allergen-sensi- myringotomy and mild ear flushing offers a very practi- tized dogs and cats, and that these CCD-specific cal, safe and efficient way to treat suppurative otitis antibodies might confound serological testing. The media in cats. objective of this study was to document the prevalence Source of funding: Self-funded. of CCD detectable in a monoclonal cocktail-based Conflict of interest: None declared. enzyme-linked immunosorbent assay (ELISA) designed for the detection of allergen-specific IgE in the sera of dogs and cats, and to define a means for successful inhibition of these CCD. The incidence of reactivity to FC-55 bromelain and a commercially available inhibitor of Efficacy of narrowband ultraviolet B therapy carbohydrate-specific antibodies (RIDA-CCD) was for eight dogs with cutaneous T-cell lymphoma evaluated in 100 dog sera samples before and after inhibition with RIDA-CCD, and a proprietary inhibitor T. HOSHINO*, M. HADA*, A. SATO*, P. F. † which contains carbohydrates derived from bromelain MOORE and M. NAGATA* (BROM-CCD). Subsequently, sera from 600 dogs and *Dermatology, Veterinary Specialists Emergency Centre, † 600 cats were evaluated using a serum diluent with and Saitama, Japan; Department of Pathology without BROM-CCD. Although both the RIDA-CCD Microbiology and Immunology, University of California, and BROM-CCD inhibitors demonstrated successful Davis, CA, USA reduction of CCD reactivity, a more efficient profile of Cutaneous T-cell lymphoma (CTCL) is a malignant inhibition was evident with BROM-CCD. Mite reactiv- skin neoplasm. In humans, ultraviolet (UV) light ther- ity in dog and cat sera was largely unaffected; however, apy is an established treatment for CTCL and chosen substantial inhibition for pollen allergens (trees, grasses as one of the first-tier treatments in early stage CTCL. and weeds) was shown. After BROM-CCD inhibition, The aim of this study was to evaluate the efficacy and 1% of canine samples and 13% of feline samples were safety of narrow band (NB)-UVB therapy for canine rendered completely negative for allergen reactivity. CTCL. Eight dogs were recruited at Veterinary Special- The results demonstrate that BROM-CCD is effective ists Emergency Centre. CTCL was diagnosed based on in reducing reactions with irrelevant carbohydrates, and typical clinical features of CTCL and histopathological that inhibition of CCD reactivity might substantially infiltrations of atypical T cells in the epidermis or der- alter the outcome of the in vitro reactivity profile used mis. The affected area was irradiated with NB-UVB for selection of allergens to be included in an using a portable device (TARNAB, Inforward, Inc.; immunotherapeutic regime. Tokyo, Japan), which supplies 312Æ11 nm centre

Source of funding: Funding for this study was provided dose of 2 µg/kg per os, once daily and 26/29 equids by Stallergenes Greer. had a dosage increase to 4 µg/kg p.o., once daily from Conflict of Interest: Four authors are employees at Stal- 0.5 to 6 years after onset of treatment. Over the course lergenes Greer. of the study, 25/29 equids died (n = 4) or were subjected to humane euthanasia (n = 21) between 1.5 and 10 years after onset of treatment. Death or euthanasia was attributed to complications of PPID, specifically FC-57 laminitis, in five equids while death in the remaining 20 equids was attributed to disorders associated with Appropriate chemotherapeutic treatment of advancing age. Eighty-six percent (25/29) of owners equine skin tumours in the field completed the follow-up survey, and 71% and 70% A. M. RASHMIR*, S. I. HONKISZ† and R. S. strongly agreed and 25% and 30% agreed that treat- MCCONNICOǂ ment with Prascend improved their equid’s quality of *Veterinary Surgical Services, Washington, DC, USA; life and prolonged lifespan, respectively. The improve- †Michigan State University, East Lansing, MI, USA; ment in clinical signs was greatest for energy level ǂLouisiana Tech University, Ruston, LA, USA (77%), hair coat (71%) and muscle mass (61%). Over- all satisfaction with treatment on a scale of 1–10 (10, Tumours of the skin are the most common form of best) was 5 (9%), 7 (9%), 8 (13%), 9 (17%) and 10 neoplasia in the horse. They are frequently treated in (52%); 87% of owners either agreed or strongly agreed field situations where the potential for public exposure that they would provide life-long treatment if they had to the chemotherapeutic agents is high. The National another equid with PPID. In conclusion, owner satis- Institute for Occupational Safety and Health (NIOSH) faction with Prascend as a treatment for PPID is high. has recently updated the requirements for handling Source of funding: Boehringer-Ingelheim Animal Health chemotherapeutic agents. Knowledge of these updated USA, Inc., Duluth, GA, USA provided Prascend for par- requirements combined with current recommendations ticipants in this study. for the use of these agents in the treatment of cuta- Conflict of interest: HS is a consultant and has received neous neoplasms will advance the treatment of the reimbursements, honoraria, and research funding from horse and provide protection from inadvertent human Boehringer-Ingelheim Animal Health, USA. exposure. Therefore, our objective is to provide accurate information regarding the treatment of the most common forms of equine tumours in the field and the appropriate management of the chemotherapeutic agents used. For example, intratumoural cisplatin has FC-59 produced favourable long-term control of several types Comparison of oral absorption of parenteral of equine skin tumours and can be performed in the triamcinolone acetonide in dogs to absorption field setting without general anaesthesia. Both the vet- of oral triamcinolone acetonide tablets, a pilot erinarian administering the cisplatin and the handler are required to wear NeoPro gloves, tyvek suits, face study shields, gloves and a N-95 mask. The drug should be C. ROMERO*, C. GRIFFIN†, M. G. PAPICH‡,G. injected with PhaSeal leur lock connectors to avoid SHEINBERG§, C. MARTIN¶ and R. HEREDIA* spray under pressure. Clean-up should be achieved in *Dermavet, Edo Mexico; †Animal Dermatology Clinic the same attire with 10% bleach solution followed by San Diego, California, USA; ‡North Carolina State 1% sodium thiosulfate solution. Being careful not to University, Raleigh, North Carolina, USA; §Centro self-contaminate, the PPE should then be disposed of Veterinario Mexico, CDMX, Mexico; ¶Vetderm, in a designated chemotherapy waste bin. Likewise, Guadalajara, Mexico treatment with other chemotherapeutic agents that are The objective of this study was to determine if inject- topical, intralesional or in impregnated beads should all able triamcinolone acetonide (TA) administered orally be considered a risk to the veterinary team and the is absorbed to the same level as TA oral tablets. A public, and protective equipment worn. comparative study was conducted with seven dogs that Source of funding: Self-funded. had not received any medications in 90 days before the Conflict of interest: None declared. study. All dogs received an oral dose of oral formulation TA (0.11 mg/kg Cortalone, Vedco, Boehringer Ingelheim; Ridgefield, CT, USA). Three weeks later, the same dogs were administered with the same oral FC-58 dose of parenteral formulation of TA (Atriben, Holli- Owner assessment of long-term treatment of day Scott; Buenos Aires, Argentina). On both occa- PPID sions, samples were taken at 0, 15, 30, 60, 120, 240 and 480 min, and 25 and 48 h post-administration of the H. SCHOTT, J. STRACHOTA and J. MARTENIUK drug, to evaluate plasma concentrations of TA by high- Department of Large Animal Clinical Sciences, College performance liquid chromatography (HPLC). CBC and of Veterinary Medicine, Michigan State University, East blood chemistry were performed at 120 min, 25 h and Lansing, MI, USA 36 h. For the analysis of data ANOVA was performed, Owners of 29 equids enrolled in a long-term safety the comparison of means was performed using the study of Prascend (1 mg pergolide mesylate tablets) for LSMEANS procedure of SAS (alpha = 0.05). There was treatment of pituitary pars intermedia dysfunction no statistical difference in plasma concentrations of TA (PPID) were surveyed about their experience with this between formulations (P = 0.05) in the measurements treatment 10 years after the start of treatment. All made at 15, 30, 60, 120, 240, 480 min, and 25 and 48 h equids enrolled in the study were initially treated with a after oral administration of oral and parenteral TA.

Comparison of means of analysed blood biometry and Source of funding: Self-funded. blood chemistry values showed no significant differ- Conflict of interest: None declared. ences (P = 0.05. These results indicate that parenteral TA can be administered by the oral route for the treatment of dog dermatopathies for veterinarians who do not have access to oral TA. FC-61 Funding source: Self-funded. Conflict of interest: none declared. Usefulness of a novel isolation and drug susceptibility test media for diagnosis and treatment of superficial bacterial folliculitis in dogs FC-60 H. SHIMOURA, M. ESUMI, S. KANDA and K. Effects of sarolaner on earthworms (Eisenia IYORI foetida) Vet Derm Tokyo, Kanagawa, Japan C. ROMERO*, R. HEREDIA*, L. MIRANDA*, G. Recently, an isolation and drug susceptibility testing SHEINBERG† and A. MARTIN‡ medium, Hifu-Quik (NIPPON ZENYAKU KOGYO, *Dermavet Hospital Veterinario, Ciudad de Mexico, Co., Ltd.; Tokyo, Japan) was made available in Japan Mexico; †Centro Veterinario Mexico, Ciudad de for dogs with staphylococcal infection. This medium is Mexico, Mexico; ‡Veterinaria Dermatologıa Veterinaria salt agar-based with four fractions containing specific Especializada, Guadalajara, Mexico concentrations of cefazolin, ofloxacin or minocycline, plus control. In this study, we aimed to evaluate the The present study examined the effect of sarolaner on usefulness of the test media for treatment of superficial earthworms. Experiment 1: three groups of rabbits were bacterial folliculitis (SBF) in dogs. Twenty-nine dogs treated with 2.5 mg/kg sarolaner (Simparica, Zoetis; with SBF were enrolled in this study. Samples were col- Parsippany, NJ, USA) and a fourth group was not lected from skin lesions and inoculated onto the four treated. Stool was collected for each rabbit group and fractions of the test media and incubated at 35 for 24– then were applied in compost sections with earthworms 48 h. Samples also were subjected to routine bacterial (three sections received sarolaner-treated rabbit faeces culture and antimicrobial susceptibility disk diffusion and one section received the control rabbit faeces). testing according to Clinical and Laboratory Standards Experiment 2: organic matter with 120 mg sarolaner Institute (CLSI) guidelines. All cases showed colony was added to three groups of 100 worms each and a formation on the control fraction of the test medium fourth group did not receive sarolaner. The morphol- within two days. In the bacterial culture test, Staphylo- ogy and number of earthworms for both experiments coccus pseudintermedius (n = 22) and S. schleiferi subsp. were evaluated on day (D)1, D15 and D30 post-treat- coagulans (n = 7) were isolated. Concordance rates ment by separating the worms from the organic matter. between antimicrobial susceptibility test results and col- Data were analysed using a Tukey’s standardized range ony formation on the test medium were 100% for cefa- test with P=0.05 and Fisher’s exact test was used for zolin (24/24) and ofloxacin (26/26), and 72.2% (13/18) statistical comparisons. For the earthworms exposed to for minocycline. All dogs were randomly treated with rabbit faeces, there were no significant differences in systemic cephalosporins (n = 13), fluoroquinolones the number of worms between sarolaner-treated and (n = 9) or tetracyclines (n = 7). Overall, the prediction control groups at D15 and D30; in fact, there was an rate of clinical response to antibiotics, based on test increase in the number of earthworms in all groups. medium results, was 86.2% (25/29). This finding For the second experiment there were no significant dif- demonstrates that this test medium could be a rapid ferences in earthworm numbers on the same evaluation and beneficial tool for determining antimicrobial sus- days. Comparison between both experiments showed ceptibility and clinical response to cephalosporins, fluo- no differences at D15 and D30 post-treatment. These roquinolones and tetracyclines in dogs with data revealed that sarolaner treatment did not affect staphylococcal SBF. the reproduction and growth of earthworms up to 30 Source of funding: Self-funded. days post-administration through either indirect expo- Conflict of interests: None declared. sure (via the faeces of rabbits medicated with sarolaner) or direct administration of sarolaner to the vermicom- post.

FC-62 Pharmacology, College of Veterinary Medicine, Auburn University, Auburn, AL, USA Real-time PCR detection of undeclared animal DNA in three commercial elimination diets for Antimicrobial resistance is a growing concern and com- plicates treatment of Staphylococcus pseudintermedius canine adverse food reactions dermatitis in dogs. Treatment with rifampicin (RFP) is T. M. S. A. BOEHM*, C. J. KLINGER† and R. S. considered only in S. pseudintermedius isolates that are MUELLER* both meticillin-resistant (MR) and multidrug-resistant *Centre for Clinical Veterinary Medicine, Ludwig (MDR). Recent data demonstrated that canine plasma Maximilian University, Munich, Germany; †Tierklinik RFP concentrations after 6 mg/kg per os are 600– Stuttgart Plieningen, Veterinary Specialist Hospital, 1,000-fold greater than the minimum inhibitory concen- Stuttgart, Germany tration [MIC (90)] of S. pseudintermedius (MIC µ An elimination diet is currently the gold standard to 0.008 g/mL). The objective of this study was to deter- diagnose canine adverse food reactions. However, mul- mine the killing properties of RFP against MR and tiple studies have shown the contamination of novel MDR S. pseudintermedius at concentrations similar to protein diets that have been claimed to be suitable for those achieved in plasma after oral dosing. One control food trials. It is unclear if several samples from differ- isolate (S. aureus ATCC 25923, American Type Culture µ ent batches are all uniformly contaminated or if such Collection; Manassas, VA, USA; MIC 0.008 g/mL) contamination only occurs sporadically and how large and three canine MR-MDR S. pseudintermedius isolates µ the percentage of undeclared protein is in contaminated (MICs 0.004, 0.008 and 0.016 g/mL) were exposed to samples. The purpose of this study was to test several RFP (MP Biomedicals; Santa Ana, CA, USA) at con- samples of different batches of three selected protein centrations from 32- to 1,024-fold greater than the iso- diets for dogs and, if present, quantify contaminating late MIC. The number of viable colony forming units proteins in those samples. Twelve samples of three was determined using the BacTiter-Glo kit (Promega; canned selected protein diets of one brand were chosen. Madison, WI, USA) at different time points (0, 2, 4, – Diets contained camel and parsnip, kangaroo and 12, 24 and 48 h) to create a time kill curve. RFP activ- pumpkin, and horse with amaranth, peach and beetroot ity against all four isolates was consistent with a time- (Terra Canis; Munich, Germany). Specimens were dependent and bacteriostatic response. At either 24 or scanned for undeclared DNA of animal origin using 48 h, each isolate demonstrated resistance to RFP at = = quantitative real-time PCR testing for horse, turkey, one (n 2) or two (n 2) of the six concentrations chicken, beef, mutton and pork antigens (Institute for tested ranging from 32- to 256-fold greater than the MIC. These resistant isolates exhibited an increase in Food Safety, Austrian Agency for Health and Nutri- tional Safety; Vienna, Austria). Eleven of 12 samples the MIC via Etest (BioMerieux; Durham, NC, USA) > µ were not contaminated with any of the tested proteins with post-exposure MIC ranging from 1 to 32 g/mL. not listed on the label. One sample with camel and These findings strongly suggest that S. pseudintermedius parsnip contained <1% turkey. When testing different develops RFP resistance at high concentrations in vitro, batches of commercial selected protein diets, contami- calling into question RFP use in dogs. nation with small amounts of proteins not listed on the Source of funding: Auburn University Department of label can occur intermittently. Whether those small Clinical Sciences Research and Graduate Studies Intra- amounts are clinically relevant is currently unknown. mural Grant. Based on this study, commercial selected protein diets Conflict of interest: None declared. may be suitable as maintenance diets for dogs with diagnosed food allergy, yet their use in diagnostic elimination diets is still questionable. Source of funding: The study was funded by Nestle FC-64 Deutschland AG. A shortened immunotherapy dose-escalation Conflict of interest: Nestle Deutschland AG was not saves time, but is it safe? A case-control study involved in the selection of food samples nor in the choice of commercial laboratory. comparing the rates of adverse reactions between conventional and fast-escalation subcutaneous immunotherapy protocols I. HERRMANN*, K. E. LOFT† and T. OLIVRY* FC-63 *Department of Clinical Sciences, College of Veterinary † Clinically relevant plasma concentrations of Medicine, NC State, Raleigh, NC, USA; MSPCA rifampicin induce resistance in multidrug- Angell Memorial Hospital, New York City, NY, USA resistant canine Staphylococcus intermedius Allergen-specific immunotherapy (ASIT) is the only group isolates intervention believed to change the course of atopic disease. As dogs appear to have fewer severe adverse † ‡ K. HICKS*, T. HATHCOCK , D. BOOTHE ,R. events (AEs) compared to people receiving ASIT, a KENNIS* and A. WHITE* prolonged dose-escalation might not be needed. Our *Department of Veterinary Clinical Sciences, College of objectives were to retrospectively compare the incidence Veterinary Medicine, Auburn University, Auburn, AL, and characteristics of AEs induced by a fast-escalation † USA; Department of Pathobiology, College of subcutaneous immunotherapy (f-SCIT) protocol to Veterinary Medicine, Auburn University, Auburn, AL, those of two conventional (c-SCIT) regimens. The AEs ‡ USA; Department of Anatomy, Physiology and during the induction phase were compared between

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 54 Abstracts three groups of 50 dogs treated with either f-SCIT pro- considered a mainstay of therapy, despite the difficulty vided by ALK-Abello ([ALK]; Round Rock, TX, USA) of administering oral tablets to cats and the potentially or with c-SCIT provided by ALK or by Stallergenes severe adverse effects of parenteral glucocorticoid injec- Greer ([Greer]; Boston, MA, USA). We graded the tions. We evaluated the efficacy and tolerability of dex- reported AEs using a modified World Allergy Organi- amethasone sodium phosphate solution 4 mg/mL zation system (local reaction to grade I–IV systemic (DSP, DexaJect SP, Henry Schein Animal Health; Oak- reactions) and assessed factors such as self-limitation. brook Terrace, IL, USA) given orally for treatment of Dogs treated with the ALK f-SCIT (11/50; 22%) and feline HD in an open-label study. Nineteen cats with Greer c-SCIT (8/50; 16%) had a significantly higher clinical signs and dermatological lesions compatible rate of total AEs compared to those receiving the ALK with feline HD were administered DSP solution at c-SCIT (1/50; 2%) (Fisher’s exact test; P = 0.004 and 0.2 mg/kg per os once daily for 21 to 28 days. Com- P = 0.03, respectively). Most of the AEs (10/11; 91%) plete blood cell counts, serum chemistry profiles and in the ALK f-SCIT group were self-limiting grade I sys- urinalyses were analysed for 6/19 cats pre- and post- temic reactions. In each of the three groups of dogs, a DSP administration. After oral DSP administration, single grade II systemic reaction was observed, and dermatological lesions evaluated through Scoring Feline there were no grade III-to-IV systemic reactions. This Allergic Dermatitis (SCORFAD) were significantly f-SCIT regimen is simpler, its maintenance phase is reduced from 7.4 (mean score) to 3.4 (54.3% reduction, reached faster, and most of its associated AEs are mild Wilcoxon’s ranked sum tests, P < 0.01) and the owner and self-limiting. Whether this f-SCIT protocol leads to pruritus Visual Analog Score (pVAS) was reduced from a faster time to efficacy needs to be determined in an initial 7.9 to 2.1 (73.9% reduction, P < 0.01). Six of future studies. 19 cats experienced mild and transient adverse effects Sources of funding: Self-funded. (sneezing, lethargy, polyphagia, inappropriate elimina- Conflict of interest: None declared. tion, and polydipsia), and a single cat developed hyper- glycaemia and glucosuria, which normalized four weeks after drug cessation. There was no reported difficulty administrating oral DSP solution. In conclusion, oral FC-65 DSP solution induced a significant and rapid improvement of clinical signs and pruritus in feline HD after Oral dexamethasone sodium phosphate three to four weeks of treatment. solution significantly reduces pruritus and Source of funding: American College of Veterinary Der- clinical lesions in feline hypersensitivity matology Resident Grant. dermatitis: an open-label study Conflicts of interest: None declared. D. MCCLINTOCK, M. AUSTEL and F. BANOVIC Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, University of Georgia, Athens, GA, USA Feline hypersensitivity dermatitis (HD) is a multifactorial, pruritic condition for which glucocorticoids are

P-1 P-2 Effect of topical medication on the The canine skin and ear microbiome: a nasomaxillary skin fold microbiome of French comprehensive survey of pathogens implicated bulldogs in canine skin and ear infections using a novel A. REXO*, B. HANSEN*, M. CLARSUND†,V. next-generation sequencing-based assay ‡ § FADOK and J. BERNSTEIN S. TANG*, A. PREM†, J. TJOKROSURJO†,M. *Dermatology and Allergy Services for Animals, † † † SARY , M. A. VAN BEL , A. RODRIGUES- Springfield, VA, USA; Division of Biotechnology, Lund HOFFMANN‡, M. KAVANAGH§,G.WU†,M. University, Sweden; ‡Zoetis-US Pet Care, Bellaire, TX, § VAN EDEN* and J. KRUMBECK* USA; Long Green Animal Dermatology Center, *MiDOG LLC, Irvine, CA, USA; †Zymo Research Baldwin, MD, USA Corp., Irvine, CA, USA; ‡Department of Veterinary Host–microbe interactions play an important role in Pathobiology, College of Veterinary Medicine & the pathogenesis of dermatitis in the nasomaxillary Biomedical Sciences, Texas A&M University, College § folds of French bulldogs, as it is often complicated by Station, TX, USA; Saddleback Animal Hospital, secondary bacterial and fungal infections. Next-genera- Tustin, CA, USA tion sequencing of DNA was used to characterize the This study analysed the complex bacterial and fungal composition of the microbiome inhabiting nasomaxil- microbiota of healthy and clinically affected canine ear lary skin folds in healthy French bulldogs. Then the and skin samples. A total of 589 canine samples were effect of two topical products on the fold microbiome included: 257 ear swab samples (128 healthy and 129 was assessed. There were seven dogs treated with an clinically affected) and 332 skin swab samples (172 enzyme-containing product effective against biofilm for- healthy and 160 clinically affected) were analysed using mation without biocidal activity; six dogs were treated next-generation sequencing (NGS) to determine both with a 2% Chlorhexidine solution with biocidal activity relative and absolute abundances of bacteria and fungi against bacteria and fungi, and six dogs were left present in the samples. This study showed that the untreated as a control for a total of 19 study partici- canine microbiota of clinically affected cases was char- pants. Dogs were assigned randomly to each group and acterized by an overall loss of microbial diversity, high the investigator was blinded. The main skin bacterial microbial biomass and overgrowth of certain members phyla inhabiting the skin fold at inclusion were Firmi- of the microbiota. The combination of relative and cutes, Actinobacteria and Proteobacteria. The main skin absolute abundances best explained the observed phe- fungal phyla were Ascomycota and Basidiomycota. notype of these samples. Compared to healthy samples, Topical treatment increased the diversity of bacterial 78.3% of the clinically affected ear samples had micro- and fungal compositions over time (increase in microbial overgrowth. Of these, 69.8% had bacterial over- bial diversity score; enzyme 42%, Chlorhexidine 11%, growth, 16.3% fungal overgrowth, and 7.0% both control <5%); these differences were statistically signifi- bacterial and fungal overgrowth. The most important cant (enzyme; P = 0.028, Chlorhexidine; P = 0.048). A microbial drivers of ear infections were Malassezia 2 clear correlation (r = 0.8) between abundance of clini- pachydermatis, Staphylococcus pseudintermedius, cally relevant pathogens and microbial diversity was S. schleiferi, and several anaerobic bacteria such as seen. We conclude from the results of this study that Finegoldia magna, Peptostreptococcus canis and Porphy- the nasomaxillary skin fold microbiome of apparently romonas cangingivalis. The anaerobic microbes identi- healthy French bulldogs contains a high abundance of fied previously were not commonly recognized as clinically relevant pathogens (mean 36.4%) and that potential pathogens in canine ear infections, perhaps topical therapy using either Chlorhexidine or enzyme due to the inclusion of these organisms within biofilms. product increases microbial diversity in the skin folds Similar observations were found for skin samples, with and reduces the relative abundance of pathogens. yeasts and anaerobes playing a less important role com- Source of funding: Self-funded. MiDOG LLC provided pared to clinically affected cases. the sample collection devices, the microbiome sequencing Source of funding: MiDOG LLC. and preliminary microbiome analysis. Conflict of interest: MK and AR-H are on the advisory Conflict of interest: MC was involved in the development board of MiDOG LLC. of the enzyme formulation however no reimbursements, fees, funding or salary were received.

P-3 Anti-inflammatory effect of shea butter extracts in canine keratinocytes in vitro D. LIM and T. OH College of Veterinary Medicine, Kyungpook National University, Daegu, Republic of Korea Shea butter (Vitellaria paradoxa) is a fat extracted from shea tree nuts and contains relatively high concentrations of nonglycerides. Shea butter extract (SBE) has

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 56 Abstracts been used to treat various skin problems in human such not to a significant degree. The wound-healing effect as burns, eczema and rashes, yet little is known about was evaluated using the scratch assay. The scratched its effect on canine skin. This study evaluated the cyto- keratinocytes were treated with 5, 10 and 100 µg/mL toxicity and anti-inflammatory effect of SBE in canine extracts. The induced lesions reduced by 13.57%, keratinocytes. The cytotoxic effects of lipopolysaccha- 74.02% and 0.42%, respectively. These results suggest ride (LPS, 5–50 ng/mL) and SBE (50–200 lg/mL) were that C. officinalis has anti-inflammatory and wound- evaluated using the Cell Counting Kit (CCK 8) assay. healing effects in canine keratinocytes and can be used Noncytotoxic concentrations of LPS and SBE were for wound repair in dogs. administered to canine cell cultures. To evaluate the Source of funding: Self-funded. anti-inflammatory effect of SBE, the levels of inter- Conflict of interest: None declared. leukin (IL)-1b, IL-8, IL-12 and tumour necrosis factor (TNF) a were measured using ELISA kits. The concentration of each cytokine was quantified in the control, LPS-treated and LPS + SBE-treated groups. Increased P-5 levels of IL-1b, IL-8 and IL-12 were found in the LPS- treated groups relative to the control groups (P < 0.05). Anti-inflammatory effects of the conditioned The LPS + SBE-treated groups showed lower levels of media from canine adipose tissue-derived IL-1b, IL-8 and IL-12 than the LPS-treated groups mesenchymal stem cells on canine (P < 0.05). These results suggest that SBE may have an keratinocytes application as a topical agent for canine inflammatory skin diseases. However, in vivo study is needed to evalu- Y. JIN and T. OH College of Veterinary Medicine, Kyungpook National ate the safety and efficacy of SBE in dogs. University, Daegu, Korea Source of funding: Self-funded. Conflict of interest: None declared. Mesenchymal stem cells (MSCs) are known to have anti-inflammatory effects and studies have shown that MSC paracrine factors are the main contributors to these effects. These paracrine factors are present in P-4 MSC-conditioned media (MSC-CM). In this study, we investigated the anti-inflammatory effects of canine Anti-inflammatory and wound-healing effects MSC-CM on canine keratinocytes. MSC-CM was made of Calendula officinalis in canine keratinocytes with canine adipose tissue-derived mesenchymal stem in vitro cells. The positive marketers of these cells identified by the flow cytometer are CD44 and CD90, and the nega- J. H. JEON, D. H. LIM, Y. S. JIN, S. G. BAE and T. tive marketers are CD29, CD34, CD45 and MHC2. H. OH Keratinocyte-conditioned media (KCM) was made with Department of Veterinary Dermatology, College of canine keratinocytes (CNT-09, CellnTec; Bern, Switzer- Veterinary Medicine, Kyungpook National University, land) as a control. Canine keratinocytes were stimu- Daegu, Korea lated with lipopolysaccharide (LPS; Escherichia coli Calendula officinalis (C. officinalis) is a traditional 026:B6) to evaluate the anti-inflammatory efficacy of medicinal plant that calms skin inflammation and pro- MSC-CM. Three groups of keratinocytes were cultured motes wound healing. In humans, the dermatological until 100% confluence was reached. The media for effects of C. officinalis have been demonstrated and Group 1 was changed to KCM for comparison with many products contain C. officinalis for wound repair. the LPS-added groups. The media of groups 2 and 3 This study aimed to evaluate the anti-inflammatory and were changed to KCM and MSC-CM with LPS added, wound healing effects of C. officinalis in canine ker- respectively. After 4 h, each group of keratinocytes was atinocytes. Petals of C. officinalis were extracted with washed and CM was re-applied. After 20 h, the concen- 70% ethanol at 85°C using a Soxhlet apparatus. To tration of pro-inflammatory cytokines, such as tumour determine the appropriate concentration of extracts, an necrosis factor (TNF) a, interleukin (IL)-1ß, IL-8 and Escherichia coli 026:B6 lipopolysaccharide (LPS) cell- IL-12 were measured using a commercial enzyme-linked viability assay was performed using a Cell Counting immunosorbent assay kit. Group 2 showed higher con- Kit (CCK 8). The final extract and LPS concentrations centrations of IL-1ß (P < 0.001), IL-8 (P < 0.05) and were 10 µg/mL and 10 ng/mL, respectively. To assess IL-12 (P < 0.001) compared with Group 1. These the anti-inflammatory effect of C. officinalis, interleukin results demonstrate that LPS can induce keratinocytes (IL)-1b, IL-8, IL-12, and tumour necrosis factor (TNF) to elevate the concentrations of some pro-inflammatory a were measured using an enzyme-linked immunosor- cytokines. Group 3 showed lower concentrations of bent assay. Statistical analysis was performed using TNFa (P < 0.001), IL-1ß (P < 0.001), IL-8 (P < 0.05) one-way ANOVA (Fisher least significant difference and IL-12 (P < 0.001) compared with Group 2. These method) with SIGMAPLOT for Windows v12.0 (Systat; results highlight the potential efficacy of MSC-CM in San Jose, CA, USA). All experiments were run in tripli- reducing skin inflammation. cate. The 10 µg/mL extracts significantly reduced the Source of funding: Self-funded. concentrations of IL-1b, IL-8 and IL-12 compared to Conflict of interest: None declared. the groups that were treated only with LPS (P < 0.05). The concentration of TNFa was decreased although

P-6 P-7 Zinc responsive dermatosis-type II syndrome Mutations in noncoding sequences of the EDA in an adult dog gene may be a cause of X-linked hypohidrotic M. CARDOSO*, P. VENTER†, C. HENRICH†,C. ectodermal dysplasia in dogs † ‡ GOMES and G. FREDO E. MOURA*, S. DALTRO†,D.SAS ‡, J. ENGRACIA *Dermalove Veterinary Dermatology Clinic, Caxias do FILHO§, M. FARIAS§ and C. PIMPAO~ § Sul, RS, Brazil; †Cao~ Noia Veterinary Hospital, Novo ‡ *Service of Medical Genetics, School of Life Sciences, Hamburgo, RS, Brazil; Diagnostic Pathology and Pontifıcia Universidade Catolica do Parana, Curitiba, Cytology Laboratory, Caxias do Sul, RS, Brazil PR, Brazil; †Instituto Goncßalo Moniz, Fundacßao~ ‡ Zinc is an important cofactor and modulator of many Oswaldo Cruz, Salvador, BA, Brazil; Genotyping – biological functions. It plays an important role in dog Diagnosticos Geneticos, Botucatu, SP, Brazil; § dermatoses, yet its deficiency is rare and caused by diet- Postgraduate Program in Animal Science, School of ary insufficiency or genetic anomalies. The goal of this Life Sciences, Pontifıcia Universidade Catolica do report is to show the potential applicability of a dietary Parana, Curitiba, PR, Brazil approach to treat type-II zinc responsive dermatosis. A X-linked hypohidrotic ectodermal dysplasia (XLHED) 2-year-old, female, mongrel dog presented with apathy, is a rare genodermatosis that affects humans, dogs, cat- symmetrical periocular hyperkeratosis, ulceration, tle, mice and probably other mammals. Affected indi- crusts and fissures on the nasal planum, paw pads and viduals are usually males with characteristic alopecia vulva. Skin cytological evaluation revealed coccoid bac- and dental abnormalities. In dogs, three mutations in teria. Amoxicillin and clavulanate at a dosage of the EDA gene have shown to cause the dysplasia. The 20 mg/kg per os, twice daily, was prescribed. After two aim of this study was to verify whether a male poodle weeks, the dermatological signs worsened and a biopsy dog with clinical and histopathological diagnosis of was obtained revealing epidermal orthokeratotic hyper- XLHED had one of these three mutations. All proce- keratosis with areas of parakeratosis, and moderate dures were approved by the Institutional Committee hyperplasia with focal areas of heavy papillomatous for Ethical Use of Animals in rResearch. The DNA of projections. Cellular debris and clusters of coccoid bac- the affected dog was submitted for next-generation teria were observed in the stratum corneum. The dog sequencing (NGS) and the result was compared to the owner reported a rancid smell in the food offered to reference sequence (CanFam3.1 NC_006621.3) and the the patient. This finding and the biopsy results sug- EDA gene sequence of a normal dog. A panel with a gested a probable dietary deficiency in zinc. A premium pool of multiplex primers was designed covering all the diet for adult dogs was prescribed (Golden Formula exons and their flanking regions in addition to the pet Small Breed Dogs, PremieR ;Sao~ Paulo, Brazil). entire intronic region from exons 4 to 8, a large part of After six weeks of the new diet, the skin lesions the 5’ untranslated region (UTR) region and the entire resolved completely. It is likely that the rancid diet pre- 3’ UTR. The NGS was performed using the Ion S5 viously eaten by the dog led to important nutrient defi- System platform. No point mutation or indels were ciencies resulting in the development of the described found in the exons and splice sites of the EDA gene, clinical signs. This case indicates that a balanced diet and a transition (approx. 779–1,188C>T) and a can be used solely to treat canine type-II zinc respon- transversion (*1,018T>A) were found in the intron 6 sive dermatosis. region and 3’UTR, respectively. Genetic diseases in Source of funding: Self-funded. humans have been reported to be caused by mutations Conflict of interest: None declared. in introns and UTRs. Therefore, the cause of the dysplasia of the dog in this study is probably a deep intronic mutation, a 3’UTR mutation or both. Source of funding: Self-funded. Conflict of interest: None declared.

P-8 induced in HDM/SEB-treated cells compared to untreated cells, including IL-4, IL-13, IL-6, IL-33, Results of intradermal skin test in 58 dogs tumour necrosis factor (TNF)a and IL-17A. In addi- with atopic dermatitis in Santiago, Chile tion, IL-31, GM-CSF, interferon-c, IL-6, IL-8, ker- B. RIVERA and V. BALAZS atinocyte chemotactic (KC)-like, monocyte Oftaderm, Clınica especializada en Oftalmologıa y chemoatractant protein-1 (MCP-1), TNFa and IL-10 Dermatologıa Veterinaria, Santiago, Chile protein levels were induced over 72 h. Taken together, these results highlight that numerous cytokines and The purpose of this study was to identify the most chemokines are induced after allergen exposure and prevalent allergens in atopic dogs in Santiago, Chile. likely play a key role in triggering clinical signs associ- Fifty-eight dogs (26 females, 32 males) with a clinical ated with canine AD. diagnosis of atopic dermatitis (AD) were tested using Source of funding: Zoetis, Kalamazoo, MI, USA. intradermal skin test (IDST). The most prevalent aller- Conflict of interest: All authors are employees of Zoetis. gens were mixture of grasses (64%) and storage mites (60%). In 25 dogs with mostly indoor lifestyle, the most common positive allergen reaction was to storage mites (69.2%). In 33 dogs living outdoors, positive reactions were most common to mixture of grasses (71.4%). P-10 Twenty-two dogs with seasonal signs of AD showed a Allergic sensitization profile from prick test in higher positive reaction to maple (66.7%). Thirty-six dogs with atopic dermatitis in Colombia dogs with permanent allergy had more positive reac- R. FERREIRA*, J. DUENAS~ †, J. GUZMAN† and J. tions to mixture of grasses (67.6%). When comparing § dogs living in urban areas with dogs living in rural SANCHEZ areas, results showed that in urban dogs the most *Dermatovet Cursos, Porto Alegre, Brasil; † § prevalent allergens were mixture of grasses (63.3%), Dermalatam, Cali, Colombia; Dermasur, Bogota, storage mites (61.2%) and Penicillium (59.2%). In rural Colombia areas the most frequent positive reactions were to mix- Skin prick test (SPT) is the first choice for the detection ture of grasses (66%), Cladosporium (66%) and maple of type I hypersensitivity in human atopic dermatitis (55.8%). These results show that environment and liv- (AD). Canine AD resembles several clinical signs of the ing conditions can influence the results of IDST. disease in the human counterpart. In canine AD, intra- Source of funding: Self-funded. dermal testing is the test of choice, and there are few Conflict of interest: None declared. reports on the use of SPT in dogs. The purpose of this study was to evaluate SPT reactions in atopic dogs and a healthy control group to six environmental allergens. The antigen panel consisted of common allergens in P-9 Colombia and more specifically house dust mites and grass pollens. Six allergen extracts were applied on the Exploring the role of cytokines and lateral thorax of forty atopic dogs (22 females, 18 chemokines in canine allergic skin disease males) and 10 healthy dogs (five females, five males). S. BOWEN, M. ALEO, S. CORUM, X. ZENG, G. The skin was pricked with a feather lancet (Duotip- RINCON and A. GONZALES Test, Lincoln Diagnostics, Inc.; Decatur, IL, USA) and Zoetis, Kalamazoo, MI, USA evaluated for the positive percutaneous reaction at 20 min post-application of the allergens. None of the Atopic dermatitis (AD) in dogs is clinically character- dogs in the control group showed a positive reaction to ized by chronic pruritis and skin lesions. It has been the allergens included in the test. In the group of aller- well-established that AD is associated with a T helper gic animals a total of 26 dogs (65%) were polysensi- cell (TH)1/TH2 cytokine imbalance, resulting in tized and 13 (32.5%) were monosensitized. Overall, the increased production of TH2 cytokines including inter- most frequently sensitizations in decreasing order were leukin (IL)-4 and IL-31, with the latter playing a key Dermatophagoides farinae (57.5%), D. pteronyssinus role in pruritus in certain animal species. The objective (40%), Blomia tropicalis (37.5%), Cynodon dactylon of this study was to characterize the role of other (22.5%) Paspalum notatum (20%) and Lolium multiflo- cytokines and chemokines in canine AD. We employed rum (17.5%). This was the first work published on a cell model utilizing peripheral blood mononuclear prick testing on dogs with AD in Colombia. cells (PBMCs) collected from beagles sensitized to a Source of funding: Self-funded. common allergen, the house dust mite (HDM; Der- Conflict of interest: None declared. matophagoides farinae) and polarized toward a TH2 phenotype. To assess gene expression within individual cells, TH2 polarized PBMCs were treated with both HDM and staphylococcal enterotoxin B (SEB) allergens for 3 to 72 h followed by 10x single cell sequencing. Eighteen different cell populations were identified with a significant increase observed for the activated memory T-cell population after 72 h. Cell type-specific gene expression of several cytokines were evaluated and compared between allergen-treated and untreated cells at each timepoint. Several cytokine transcripts were

P-11 time PCR was done for IL-31 gene expression. One dog was removed on D8 due to severity of flare. Pruritic area and lead time after food Another dog was removed from analysis as an outlier. challenge test L-amino-acid oxidase-like gene expression increased (5- † fold changes on D1 and D8, 4.5-fold on D15 and 2- K. KAWANO* and H. SHIMAKURA > *Tokyo Animal Allergy Center, Adachi-Ku, Tokyo, fold on D28). Gap-junction protein had 3-fold change † increases on D1, D8 and D28. Calumenin had 2.5-fold Japan; Department of Veterinary Microbiology, School > of Veterinary Medicine, Azabu University, Sagamihara- increase on D1 and D8. Keratin-5 had 3-fold increase shi, Kanagawa, Japan on D8. Trichohyaline and keratin-associated-proteins gene expression had 3.3- to 4-fold change decrease at Elimination diet trial and food challenge testing remain D15 and D28. Compared to D0, IL-31 gene expression the gold standards for diagnosis of cutaneous adverse was significantly increased on D8 (P = 0.0098), D15 food reactions in dogs. These tests are difficult to per- (P = 0.0068) and D28 (P = 0.0187). Correlation of IL- form due to the risks involved with food challenge test- 31 mRNA and clinical scores was not significant ing. We reported the characteristics of food challenge (R = 0.33; P = 0.078). It is concluded that allergen testing, aiming to promote it in general veterinary prac- challenge decreases gene expression of markers of kera- tice. Forty-six dogs with cutaneous adverse food reac- tinization and increases keratins associated with prolif- tions were enrolled in this study. Elimination diet trial eration. Gene expression of IL-31 is increased in was performed for four weeks using food composed of chronic allergen exposure. novel proteins. Subsequently, the dogs were fed the Source of funding: Self-funded. food that had previously been eaten during food chal- Conflict of interest: None declared. lenge testing. The pruritus Visual Analog Scale score was recorded before and after the elimination diet trial. The time and the site of onset after food challenge testing were recorded. Significant symptomatic improve- P-13 ment was observed after the elimination diet trial (P<0.0001). All dogs exhibited pruritus and none Filaggrin expression in the skin of normal and exhibited anaphylaxis after the food challenge testing. atopic dogs challenged with house dust mites Among these dogs, 60.9% developed pruritus within R. MARSELLA, K. AHRENS, R. WILKES, N. S. 12 h; notably, 23.9% of dogs developed pruritus in 3– MUNGUIA and M. DORR 6 h. The limbs exhibited the highest frequency of pruri- Department of Small Animal Clinical Sciences, College tus (56.5%) followed by the face (26.1%). The present of Veterinary Medicine, University of Florida, study is the first to investigate the detailed status after Gainesville, FL, USA food challenge testing in dogs. These results will assist the veterinarians and the owners in the food challenge Filaggrin and filaggrin-like protein (filaggrin 2) have decision-making process. been described in dogs. This study aimed to investigate Source of funding: Self-funded. the expression of these proteins in normal and atopic Conflict of interest: None declared. dogs, and to investigate the effect of allergen exposure in both populations. Ten atopic beagles and five normal research beagles were challenged daily with allergen (Dermatophagoides farinae) for three days. Biopsies P-12 were taken before allergen exposure and after the third exposure [Day (D)3] in both groups. Biopsies also were Microarray analysis of differential gene taken from atopic dogs on D10, a week after last aller- expression in atopic beagles undergoing acute gen exposure. Immunohistochemical evaluation was and chronic allergen challenges performed for both filaggrins. Five pictures of each sec- † tion were imaged, then scored by four observers (una- R. MARSELLA*, A. RIVA , K. AHRENS*, R. ware of group and time) for intensity of staining and WILKES*, N. S. MUNGUIA* and M. DORR* epidermal thickness. A mixed-effects model indicated *Department of Small Animal Clinical Sciences, College that, upon allergen exposure, the intensity of filaggrin of Veterinary Medicine, University of Florida, † staining was significantly increased in the atopic dogs Gainesville, FL, USA; Interdisciplinary Center for compared to the normal dogs (P = 0.01). Atopic sam- Biotechnology Research and Bioinformatics Center, ples also showed a significant increase in epidermal University of Florida, Gainesville, FL, USA thickness after allergen exposure (D3 versus D0, This study aimed to investigate differential gene expres- P = 0.005) while the normal dogs did not show change. sion of chronic reactions as prior studies have only There were no significant differences between normal examined acute reactions. Five atopic beagles allergic and atopic dogs in the staining appearance for filag- to Dermatophagoides farinae were epicutaneously chal- grin-like protein in the stratum granulosum and cor- lenged daily for three days and twice weekly for three neum. In summary, atopic dogs appear to be more more weeks. Biopsies were taken before [Day (D)0] and reactive to allergen exposure than normal dogs based 24 h after allergen exposure (D1, D2, D8, D15 and on increased epidermal thickness and filaggrin expres- D29). Dermatitis was scored those days using the sion. Further studies should address the potential dif- Canine Atopic Dermatitis Extent and Severity Index, ferences in function between these two filaggrins as 3rd iteration (CADESI-03). Total mRNA was they pertain to the skin barrier function. extracted, complementary DNA was made and hybri- Source of funding: Self-funded. dized to a Canine Genome 2.0 Array (Genechip Conflict of interest: None declared. Hybridization Kit, Affymetrix; Santa Clara, CA, USA). The chip did not include interleukin (IL)-31 so real-

P-14 Yorkshire terriers were most likely to present cAD (OR: 3.246, 95% CI: 1.301, 8.104; P = 0.008), French Flea or not flea? Portuguese water dog atopic bulldogs FA (OR: 2.622, 95% CI: 1.444, 4.760; dermatitis phenotype P = 0.001) and pugs both CAD with FA (OR: 2.922, 95% CI: 1.035, 8.282; P = 0.035). Age was shown to be T. CASEMIRO, H. L. PEREIRA, T. NUNES, E. a risk factor, with more animals diagnosed with CAD DELGADO and A. M. LOURENCßO at 3–6 years of age (OR: 1.813; 95% CI: 1.261, 2.605; Centre for Interdisciplinary Research in Animal Health, P = Faculty of Veterinary Medicine, University of Lisbon, 0.001), and at six months to 3-year-old for animals with FA (OR: 3.198; 95% CI: 2.051, 4.988; Lisbon, Portugal P = 0.0001). The group with both diseases showed no Dogs with flea allergy dermatitis (FAD) typically have age predisposition. Animals with FA also exhibited an lesions distributed on the dorsolumbar area. Many Por- increased risk factor for developing otitis (OR: 1.891; tuguese water dogs (PWDs) with atopic dermatitis 95% CI: 1.175, 3.043; P = 0.008), compared to animals (AD) appear to have lesions in this area as well. The with cAD. The presence of cAD and food allergy in objective of this study was to establish if the dorsolum- dogs in this study is similar to what has been reported bar area could be included in the PWD AD phenotype. previously, with a higher risk factor in French bulldogs Ninety-nine client or breeder-owned PWDs were and Yorkshire terriers. enrolled and AD diagnosis was made in 26. Clinical Source of funding: Self-funded. diagnosis of AD was made according to accepted crite- Conflict of interest: None declared. ria and after ruling out other causes of pruritus. Serol- ogy for flea saliva-specific IgE antibodies was performed (LETIPharma; Barcelona, Spain) and body areas showing lesions were recorded for each dog. P-16 Results were compared with a generic population of atopic dogs using the Z-test for proportions. Results Durability of the atopic dermatitis phenotype showed that PWDs with AD have a special predisposi- in a laboratory colony of beagles tion to lesions on the dorsolumbar area not related to K. RUDOLPH, T. R. YELLOWHAIR, J. J. FAD (38.5%; P < 0.05). Additionally, significant differ- DEARING and E. G. BARRETT ences were found in the percentage of atopic PWDs Lovelace Biomedical Research Institute, Albuquerque, showing lesions in certain areas compared with the gen- NM, USA eric population of affected dogs: front feet (96.2%; P < 0.05), pinnae (92.3%; P < 0.001), eyelids (69.2%; Lovelace Biomedical Research Institute (Albuquerque, P < 0.001), development of otitis (92.3%; P < 0.001), NM, USA) has been maintaining a colony of allergi- dry skin (69.2%; P < 0.001) and conjunctivitis (57.7%; cally sensitized beagles for the past 20+ years. Herein P < 0.001). Dry skin may have resulted from a primary we characterized the durability of the phenotype of this defect. In conclusion, the PWD is predisposed towards unique colony of dual-sensitized [ragweed (RW) and development of lesions on the dorsolumbar area. This house dust mite (HDM)] beagles and their response to can be misleading as this region is classically associated steroid treatment. The colony was established by breed- with FAD. ing only allergically sensitized females and then chal- Source of funding: Self-funded; Flea-specific IgE serology lenging their offspring to a series of subcutaneous, testing was funded by LETIPharma, Barcelona, Spain. epicutaneous and inhalation challenges with crude Conflict of interest: None declared. HDM and RW extracts (Greer Laboratories; Lenoir, NC, USA) beginning within the first week after birth and continuing until 16 weeks of age. Subsequently, dogs were challenged by epicutaneous HDM or inhaled P-15 RW to induce either a dermatitis or asthmatic-like response while receiving five daily pre-treatments of First report of allergic disease distribution oral steroid at 1 mg/kg (Prednisolone, Lloyd, Inc.; dogs in a dermatology specialty hospital in Shenandoah, IA, USA). The dermatitis phenotype is Mexico City characterized by an increase in the clinical modified Canine Atopic Dermatitis Extent and Severity Index A. ROMERO*, G. SHEINBERG*, C. ROMERO† and § (CADESI) scoring of papules, excoriation and ery- A. M. CORDERO thema, and an increase in pruritus, as measured by *Centro Veterinario Mexico, Mexico City, Mexico; † § video recording of pruritic behaviour. The dermatitis Dermavet, Estado de Mexico, Mexico; Vetderm, phenotype is attenuated by prophylactic treatment with Guadalajara, Mexico oral steroids. By contrast, the asthmatic phenotype is Canine dermatological allergic manifestations are com- only partially responsive to oral steroid treatment with mon causes of veterinary consultation. Canine atopic a reduction in lung eosinophils and lack of effect on dermatitis (cAD) and food allergy dermatitis (FA) are lung neutrophilia and bronchoconstriction. This unique considered clinically indistinguishable, requiring food allergically sensitized beagle colony exhibits both a trials for diagnosis. Several studies have attempted to robust complex dermatitis and asthmatic phenotype describe the affected dog populations. The present that shares many features similar to both clinical dis- study evaluates the presence of cAD, FA or both disease in companion pets and humans with differential eases in a population of dogs treated in Centro Veteri- responsiveness to steroids. nario Mexico. Retrospective data of 368 diagnosed Source of Funding: Lovelace Biomedical Research Insti- allergic dogs were obtained from August 2018 to Octo- tute, Albuquerque, NM, USA. ber 2019. Of patients, 58.4% presented cAD, 27.7% Conflict of interest: Authors are employees of Lovelace FA and only 13.8% were diagnosed with both. Biomedical.

P-17 Source of funding: European Society of Veterinary Der- matology Practitioner Grant 2012. Expression of canine b-defensin 103 mRNA in Conflicts of interest: None declared. skin biopsies from atopic and healthy dogs after exposure to different bacteria P-18 C. SIMOU*, K. KOKKINAKI†, M. CHATZIS†,L. ARCHER‡, V. NTAFIS§, E. VINGOPOULOU¶,V. Evaluation of skin prick test to identify SIARKOU¶, P. XENOULIS†,M. sensitization to house dust mite allergens in SARIDOMICHELAKIS† and D. SANTORO‡ dogs † *Small Animal Clinic, Athens, Greece; Clinic of V. CUNHA*,†, C. SOUZA‡, E. CONTRERAS§,T. Medicine, Faculty of Veterinary Science, University of † † ‡ ‡ NUNES , R. ROCHA and F. BERNARDES Thessaly, Karditsa, Greece; Department of Small A/V Clinica Veterinaria, Rio de Janeiro, RJ, Brazil; Animal Clinical Sciences, College of Veterinary †FDA Allergenic, Rio de Janeiro, RJ, Brazil; Medicine, University of Florida, Gainesville, FL, USA; ‡ § Department of Veterinary Clinical Medicine, University Faculty of Animal Science and Aquaculture, of Illinois, Urbana, IL, USA; §Department of Clinical Agricultural University of Athens, Athens, Greece; ¶ Sciences, Colorado State University, Fort Collins, CO, Laboratory of Microbiology and Infectious Diseases, USA Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, Thessaloniki, Greece Prick tests have been used in human medicine for dec- ades for the evaluation of skin sensitization towards Defective production, activity, or both, of host defence environmental allergens. The objective of this study peptides are postulated to be a cause of staphylococcal was to evaluate the prick test to identify sensitization skin infections in dogs with atopic dermatitis (AD). b to house dust mites (HDM) in dogs with clinically diag- The aim of this study was to compare canine -defensin nosed atopic dermatitis (AD). Ten healthy dogs and 45 103 (cBD103) mRNA expression between dogs with dogs with AD were enrolled. Histamine and saline solu- AD and healthy controls, after incubation of skin biop- tions were used as positive and negative controls, sies with a meticillin-susceptible or a meticillin-resistant respectively. Three solutions of Dermatophagoides fari- Staphylococcus pseudintermedius clinical isolate (MSSP nae (DF), D. pteronyssinus (DP) and Blomia tropicalis and MRSP, respectively) and a normal member of the (BT) (FDA Allergenic; Rio de Janeiro, RJ, Brazil) were skin microbiota (Kocuria rhizophila; formerly Micrococ- tested. Prick tests were performed in duplicate accord- cus luteus). One 8 mm punch biopsy was obtained from ing to Duotip-II manufacturer recommendations (Lin- the noninfected skin of the abdominal area from 10 coln Diagnostics Inc.; Decatur, IL, USA). A positive dogs with AD and 10 healthy controls. The sample was response to an antigen was defined as wheals having a sectioned into four equal-sized pieces and each incu- ≥ ° mean diameter of 3 mm larger than the mean diame- bated at 37 C with either phosphate buffer saline (PBS) ter of negative control wheals. One of the 10 healthy or one bacterial (MSSP, MRSP or K. rhizophila) sus- 8 dogs had a positive response on the prick test whereas pension (2 x 10 colony forming units/mL) for 24 h. 30 of the 45 dogs with AD had a positive response. Biopsy samples were then washed in PBS; epidermis – ° The odds of having a positive antigen response on the was trimmed off the dermis and stored at 80 C until prick test were greater among dogs with AD (OR 18.0; measurement of cBD103 mRNA with quantitative 95% CI, 2.1 to 155.6; P = 0.002). Of the 30 positive reverse transcription PCR. cBD103 mRNA was unde- dogs with AD, three had positive responses to one tectable in most samples. There was no difference allergen (1 DF, 2 DP). Eighteen to two different mites’ between dogs with AD and healthy controls or between allergens (10 DF and DP, six DF and BT, two DP and PBS, MRSP, MSSP and K. rhizophila regardless of BT). Nine of the 30 dogs had positive responses to all whether all samples or only the samples with at least three mites’ allergens. Duotip-II prick test is a poten- one positive result were analysed. In conclusion, under tially safe tool to evaluate HDM sensitization in dogs these experimental conditions, the incubation of atopic with AD. or healthy canine skin with different bacteria failed to Source of funding: Self-funded. upregulate the expression of cBD103 mRNA. Conflicts of interest: None declared.

P-19 atopic dogs were included and randomized in a 1:1 ratio to a standard of care or a weekly e-mail contact Topical therapy increases the efficacy of group. Control visits were performed after three and lokivetmab in canine atopic dermatitis and six months for evaluation of skin lesions (Canine Ato- delays disease flares: a single-blinded, pic Dermatitis Lesion Index) and pruritus scoring randomized, controlled study (Visual Analog Scale) and for compliance estimation. † Six dogs were lost to follow-up and 24 dogs completed E. BENSIGNOR* and E. VIDEMONT the study. Greater compliance was observed in the *Veterinary Dermatology Referral Service, Paris, intervention group (62% versus 34% after three France; †Veterinary Hospital, Annecy, France months, and 54% versus 31% after six months; Lokivetmab is an effective treatment for canine atopic P < 0.05). CADLI and pruritus decreased more in the dermatitis (cAD). The aim of this prospective study intervention group at all time points (P < 0.05). This was to evaluate the effect of combining the administra- study underlines the challenge of long term compliance tion of lokivetmab with a topical therapy to better to treatments in canine atopic dermatitis. Compliance is improve cAD skin lesions and extend the duration of low in the short term and decreased in the long term, the efficacy of the injectable treatment. Thirty atopic yet can be increased by a close follow-up. This is a con- dogs were included and randomly allocated to one of siderable challenge for both general practitioners and two treatment groups: A, a single injection of lokivet- veterinary dermatologists. mab (Cytopoint, Zoetis; Louvain la Neuve, Belgium); Source of funding: Self-funded. or B, a single injection of lokivetmab combined with a Conflicts of interest: None declared. weekly topical therapy of a spot-on (ATOP 7, Der- moscent, LDCA; Castres, France) and a shampoo (ATOP 7; Dermoscent, LDCA), specifically designed to improve atopic skin barrier defect. Clinical parameters P-21 evaluated included pruritus (Visual Analog Scale), skin Use of a probiotic blend in 42 dogs diagnosed lesions using Canine Atopic Dermatitis Lesion Index with canine atopic dermatitis associated with (CADLI), cosmetic evaluation and owner and investigator global assessment of efficacy (OGATE). Dogs recurrent superficial bacterial infection were followed-up at Day (D)10, D17 and D31 and until M. CARDOSO* and F. OLIVEIRA† a clinical relapse. An improvement was noted for all *Dermalove Veterinary Dermatological Office, Caxias dogs as soon as day 10 without difference between the do Sul, RS, Brazil; †Formula Bichos Veterinary two groups. At D17 and D31, CADLI, cosmetic and Manipulation, Novo Hamburgo, RS, Brazil OGATE scores were significantly more improved in Probiotics are live bacteria that, when ingested, may < – Group B than in Group A (P 0.05, Wilcoxon have beneficial effects in humans or animals. The main – Mann Whitney U-test). A longer lasting remission also goal of this study was to test the efficacy of a probiotic was noted in Group B compared to Group A. Dogs for control of recurrent bacterial skin infections, a com- being treated by the combination treatment showed a mon condition in dogs with atopic dermatitis (cAD). longer extended time to flare compared to dogs treated For this study, a probiotic blend was used that con- < with lokivetmab alone (P 0.05). This study suggests tained the following: Lactococcus lactis, Lactobacillus that the combination of lokivetmab with topical thera- casei, Lactobacillus salivarius, Lactobacillus acidophilus pies is of great value for treating cAD. and Bifidobacterium bifidum (Formula Bichos; Novo Source of funding: Laboratoire de Dermo-Cosmetique Hamburgo, RS, Brazil). Forty-two patients with cAD Animale (LDCA). and recurrent bacterial infection, of both sexes, differ- Conflict of interest: Both authors are consultants for ent breeds and weights, that used the probiotic blend at Zoetis Animal Health and LDCA. a dose of 8.04 x 108 colony forming units/kg for ≥90 days in a row were monitored for two years. Adminis- tering the probiotic blend while fasting in the evening improves the adhesion of strains to the intestine. P-20 Patients were monitored for nine months after starting Long-term adherence to topical treatments in supplementation through clinical exams and skin cyto- canine atopic dermatitis: a randomized logical evaluation once monthly. Of the 42 patients, 14 (33.3%) did not respond to treatment and a new sys- intervention study temic antibiotic cycle was administered and 28 (66.6%) E. BENSIGNOR*, M. LARHER* and J. PATTYN† showed a positive response and the infection did not *Veterinary Dermatology Referral Service, Paris, return. Twenty of the 42 (47.6%) needed topical treat- France; †Veterinary Clinic, Cesson Sevigne, France ment with associated antibacterial shampoo, six Estimates from the World Health Organization indicate (14.3%) needed topical and systemic antipruritic treat- that patients in developed countries only take about ment, and two (4.8%) systemic antipruritic treatment. 50% of prescribed medicine for chronic diseases such It was observed in two (4.8%) that when probiotic as hypertension and diabetes. It may be the same for blend was suspended, the bacterial infection returned owners of atopic dogs. The aim of this randomized within 14 days. Using these probiotics appears to aid in intervention study was to evaluate the value of a close the control of recurrent bacterial infection in cases of follow-up monitoring programme to improve the com- cAD as systemic antibiotics are reduced. pliance of pet owners for treating their dog. Thirty Source of funding: Self-funded. Conflict of interest: None declared.

P-22 formulation of CsA (Cyclavance, Virbac; Carros, France) is now available for dogs in many countries Investigation of the efficacy of a topical k- and it facilitates precise dosing and owner compliance. opioid agonist in a model of canine atopic The purpose of this study was to evaluate the safety dermatitis and efficacy of this CsA for the control of cAD. Thirty-nine client-owned dogs with nonseasonal cAD R. MARSELLA*, K. AHRENS*, R. WILKES*, A. † † received an oral solution of Cyclavance (5 mg/kg), once TRUJILLO*, M. SOEBERDT and C. ABELS daily for 60 days. Before treatment and at Day (D)30 *Department of Small Animal Clinical Sciences, College and D60 after therapy, skin lesions were graded by clin- of Veterinary Medicine, University of Florida, † icians using the Canine Atopic Dermatitis Extent and Gainesville, FL, USA; Sudbrackstrasse, Bielefeld, Severity Index, 4th iteration (CADESI-04). Pruritus Germany was assessed by the owners using a Visual Analog Scale The aim of this prospective, four week, placebo-con- (pVAS). The percentage of dogs showing reduction of trolled, cross-over study was to investigate efficacy of pruritus from baseline on D30 and D60 was 44.27% 1% topical k-opioid agonist gel in a model of canine and 60.85%, respectively, and the CADESI-04 score atopic dermatitis (AD). Fourteen beagles were chal- decreased by 50% and 69.84%, respectively. The clini- lenged by epicutaneous application of house dust mites cal improvement reported was 87.2% of the cases. A every three to four days for a total of nine challenges. statistically significant improvement was seen in the Severity of dermatitis was assessed (Canine Atopic Der- dog’s quality of life (P < 0.001). Haematological and matitis Extent and Severity Index, 3rd iteration) before serum chemical mean values remained within the nor- the first challenge (baseline) and 6 h post-allergen chal- mal reference ranges. Mild gastrointestinal signs were lenge. Pruritus was monitored using HERO cameras seen in 15% of the dogs. Our study provides evidence (GoPro; Santa Barbara, CA, USA) that recorded epi- that Cyclavance is safe and reduces the severity of clini- sodes of pruritus over a 75 min period. Pruritus scoring cal signs in dogs with nonseasonal cAD. Thus, Cycla- was evaluated at 10 time periods [baseline, 4 h post- vance should be considered a valuable alternative allergen challenge and the last day of the study on Day therapy in dogs with cAD. (D)28]. Scoring was done blindly by personnel using Source of funding:Dermatology Research grant from Vir- BORIS software (DBios; Torino, Italy) for 60 min after bac. the dogs had settled for 15 min. A global subjective Conflict of interest: RL received reimbursements, fees score was given using Visual Analog Scale. A four week and funding from Virbac for this study. washout period occurred, dogs were crossed-over and the study was repeated. Topical k-opioid agonist gel (Wolff GmbH; Bielefeld, Germany) was applied once daily on the inguinal area (0.6 mL/dog). The results P-24 were analysed using combined data. ANOVA showed significant effect of time (P < 0.0001) and group Evaluation of a new ciclosporin A compared to (P = 0.0001) on dermatitis scores. Overall, no statisti- prednisolone for treatment of canine atopic cally significant effect on pruritus was found owing to dermatitis in a randomized controlled trial a crossing of scores on D17. Over time, the placebo ‡ † R. LUCAS*, M. C. A. TEIXEIRA , J. LOPES ,S.Z. scores increased while the active ingredient showed ‡ ‡ § MACHADO , M. D. LUSTOZA , P. BRIANTAIS , decrease after the first three weeks. It is concluded that D. BEVIANI*, T. M. ALBUQUERQUE*, C. this approach is promising in dogs with AD and longer PELEGRINI* and V. M. C. CUNHA* studies with more frequent application may be benefi- *Dermatoclinica, Sao~ Paulo, SP, Brazil; cial. † ‡ Dermatoclinica, Sao~ Paulo, SP, Brazil; Virbac, Sao~ Source of funding: Wolff GmbH & Co. KG Arzneimittel. § Paulo, SP, Brazil; Virbac, Carros, France Conflict of interest: RM has received grant funding from Wolff for the completion of this project and MS and CA Ciclosporin A, a calcineurin inhibitor, has been used are employees of Wolff GmbH & Co. KG Arzneimittel. for the long-term treatment of canine atopic dermatitis (cAD). A new oral liquid formulation of ciclosporin A (Cyclavance, Virbac, Carros, France) is now available for dogs and it facilitates precise dosing and owner P-23 compliance. The purpose of this study was to compare the efficacy of Cyclavance with prednisolone in the Evaluation of safety and efficacy of a new treatment of cAD and evaluate the possible adverse ciclosporin A in the therapy of naturally events from each therapy. Ninety-four dogs with non- occurring canine atopic dermatitis seasonal cAD were randomly allocated to receive an ‡ † oral solution of either Cyclavance (5 mg/kg daily) or R. LUCAS*, S. Z. MACHADO , J. LOPES ,M.C. prednisolone (Prederm, Ourofino Saude Animal; Cravi- A. TEIXEIRA‡, M. MASCARENHAS†,M.D. ‡ § nos, SP, Brazil) at 1 mg/kg once daily for 15 days, then LUSTOZA , P. BRIANTAIS , D. BEVIANI*, T. M. once every two days for 15 days, and then once every ALBUQUERQUE*, C. PELEGRINI* and V. M. C. three days hours for 30 days. Dogs treated with pred- CUNHA* nisolone manifested more adverse events when com- ~ † *Dermatoclinica, Sao Paulo, SP, Brazil; Promove, Rio pared to ciclosporin. Pruritus Visual Analog Scale score de Janeiro, RJ, Brazil; ‡Virbac, Sao~ Paulo, SP, Brazil; § and Canine Atopic Dermatitis Extent and Severity Virbac, Carros, France Index, 4th iteration decreased by 58.9% and 54.1%, For several years ciclosporin A (CsA) has been recog- respectively, in the prednisolone group and by 49.2% nized as a major component of canine atopic dermatitis and 65.7%, respectively, in the Cyclavance group. The (cAD) multimodal therapy. A new oral liquid veterinarians who oversaw the trial reported a clinical

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 64 Abstracts improvement in 89.1% (prednisolone) and 86.4% P-26 (Cyclavance). According to the owners, quality-of-life improvement after 60 days of treatment ranged from Sublingual immunotherapy (SLIT) is beneficial 36.6% to 93.5% in the dogs treated with prednisolone. in long-term atopic Shiba dogs in Japan: two By contrast, animals treated with Cyclavance showed a case reports 70.5–95.5% improvement by the end of treatment. The M. IMAMOTO*, S. IMAMOTO*, Y. FUKUI† and K. present findings demonstrate the effectiveness of Cycla- ‡ vance for the treatment of cAD, resulting in improved YAMADA *Shinjo Animal Hospital, Nara, Japan; †Komachi QoL and fewer adverse events than prednisolone. ‡ Source of funding: Dermatology Research grant from Animal Hospital, Ibaraki, Japan; Yamada Animal Virbac. Hospital, Tokyo, Japan Conflict of interest: DL received reimbursements, fees Sublingual immunotherapy (SLIT) is a therapeutic and funding from Virbac. option in canine atopic dermatitis (cAD). The Shiba breed in Japan is predisposed to cAD, yet the efficacy of SLIT remains unknown in atopic Shiba dogs. This study describes the treatment of two Shiba dogs in P-25 Japan with SLIT. The first case was a 10-year-old Shiba male, who had four years of therapy with pred- Control of pruritus in atopic dogs with nisolone combined with antihistamines, prednisolone lokivetmab in Rio de Janeiro, Brazil combined with ciclosporin, oclacitinib, and a combina- R. RAMADINHA and J. ALMEIDA tion of oclacitinib and ciclosporin without significant Animalia Clinica Veterinaria, Rio de Janeiro, Brazil improvement. SLIT (Diamond Animal Health Inc.; Des Moines, IA, USA) containing dust mites based on Itching is an uncomfortable sensation that induces self- serum immunoglobulin (Ig)E testing was started. For mutilation in atopic dogs. There are several medications the first two years of SLIT, prednisone and ciclosporin to control itching and some have undesirable adverse support was provided. Six months after the initiation of effects. Lokivetmab (Cytopoint, Zoetis; Kalamazoo, the SLIT treatment, the patient recovered from the skin USA) was chosen because it can control pruritus in ato- lesions and the alopecia. The second case was a 7-year- pic dogs with minimal to no adverse effects. Between old female showing seasonal allergic pruritus around June 2019 and February 2020, 46 dogs diagnosed with the eyes and muzzle from summer to autumn. The first canine atopic dermatitis (cAD), of different breeds and clinical signs were treated with prednisolone and anti- ages, were treated at Animalia Clinica Veterinaria. Most histamines. Because of the marginal effect, the patient dogs were <7-year-old (63.0%) and shih tzus were more was switched to prednisolone and oclacitinib, and clini- Malassezia Staphylo- frequent. All animals with spp. or cal signs worsened. The serum IgE test revealed grass coccus spp. infections were treated previously with 3% (perennial rye and timothy) and weed (sweet vernal) chlorhexidine shampoo, antifungal (itraconazole) or reactivity. SLIT was started while the patient continued antibiotic (cefalexin) as needed. Pruritus was treated with – prednisolone and oclacitinib treatment. After a few prednisolone 0.5 mg/kg orally for 10 15 days, with months, the pruritus decreased, and after three years of regressive doses. Once the infections resolved and pruri- SLIT treatment, it was possible to decrease the dose of tus was controlled and they were off therapy for five to15 systemic treatments. No adverse effects were observed days, the dogs were administered lokivetmab subcuta- in either case from the sublingual treatment. It is con- neously at a dose of 2 mg/kg. One dog presented sterile cluded that the SLIT treatment was beneficial in these pustular dermatitis three days post-administration, with two Shiba dogs. spontaneous resolution. The pruritus was controlled in Source of funding: Self-funded. 80.4% (37/46) of the animals and 32/37 of these main- Conflict of interest: None declared. tained control of their cAD with repeat dosing of lokivetmab. Five owners considered the results to be excellent, yet lacked financial ability to maintain control. Three owners preferred to return to previous therapy with oclacitinib. In conclusion, lokivetmab is considered an P-27 effective therapy for cAD. Hypoallergenicity of an insect-based diet in the Source of funding: Self-funded. treatment of canine cutaneous adverse food Conflict of interest: None declared. reaction K. LEE, T. YUN, Y. KOO, M. YANG and B. KANG Laboratory of Veterinary Internal Medicine, Veterinary Teaching Hospital, Chungbuk National University, Cheongju, Republic of Korea The objective of this study was to evaluate the beneficial effect of short-term use of an insect-based diet in dogs with cutaneous adverse food reaction (CAFR). Nineteen dogs with CAFR were enrolled and classified into three groups. The insect-based diet group (n = 7) and the salmon-based diet group (n = 6) were fed with mealworm- and salmon-based diets, respectively, and the control group (n = 6) was given commercial diet for 12 weeks. In all groups, the skin lesions were evaluated based on the Canine Atopic Dermatitis Extent and

Severity Index, 4th iteration (CADESI-04). Transepi- significantly lower when compared with the control dermal water loss (TEWL) was measured to assess skin group at eight weeks (P = 0.008) and 12 weeks barrier function. The pruritus level and alteration of (P = 0.012). The TEWL of control group was signifi- medication usage also were evaluated. All indices were cantly higher than that of insect-based diet group at 12 assessed every four weeks after the initial administra- weeks (P = 0.022) and that of salmon-based diet group tion of diets. In the pruritus level and medication at every time-point. These results demonstrate a poten- scores, there were no significant between- and within- tial hypoallergenicity of the insect-based diet. group differences at all time-points. However, in the Source of funding: Basic Science Research Program insect-based group, a significant decrease in the through the National Research Foundation of Korea CADESI-04 score was observed at eight weeks com- (NRF) funded by the Ministry of Education pared to that of the baseline score (P = 0.031). The (2017R1D1A3B03028863). CADESI-04 score of the insect-based group was Conflict of interest: None declared.

P-28 of canine scrotal nodules where the infiltrate was characterized with specific histiocytic markers. Skin biopsies Probable pemphigus foliaceus in a 4-month-old from scrotal multiple nodules were obtained for kitten due to a vaccine histopathological evaluation. Periodic acid Schiff (PAS) N. A. MCEWAN stain and immunofluorescence were performed by com- University of Liverpool, School of Veterinary Science, bined use of three markers: Iba-1 for histiocytes (ab- Liverpool, Cheshire, UK cam, ab5076), Macrophage Marker (CD11b, CD68, CD163, CD14, CD16) antibody Panel (abcam, The purpose of this report is to detail pemphigus foli- ab254013) and vimentin (SanBio, MONX10662). aceus (PF) in a 4-month-old kitten associated with vac- Merged images were used to localize and co-localize the cinations and to emphasise that some cats will respond positive signals. All cases were negative for PAS stain better to dexamethasone than prednisolone. Four days and showed mixed immunoreactivity to the three anti- after routine vaccination, the patient showed crusting bodies. Most of the infiltrating cells were Iba-1 positive; skin lesions on the face, feet and a caseous paronychia. the macrophage antibody panel immunostained many, There also was hair loss and devitalisation of the skin not all, of the Iba-1 positive infiltrate thus documenting of the tail tip. Grossly the skin lesions were consistent the presence of a double population. Vimentin was with PF. The lesions on the tail tip were suggestive of localized around the nodules in spindle-shaped cells vasculitis. Cytological results and skin biopsies were and also co-localized with the other infiltrating cells. consistent with PF. The patient previously had been Immunomodulatory therapy was ineffective in two dogs treated with prednisolone per os at 5.3 mg/kg/day with with lesions spontaneously regressing; one dog without no improvement, so this was changed to dexametha- therapy had stable lesions. This study documents that sone p.o. at 0.5 mg/kg/day. Marked improvement was at least a double population of histiocytic cells was pre- seen after 14 days and then the dose was tapered and sent within the scrotal nodules. stopped at the end of three weeks. A six month follow- Source of funding: Local institutional funding, University up did not show any recurrence. Most cases of PF are of Pisa. idiopathic and typically are a life-long condition. In this Conflict of interest: None declared. case the condition resolved and did not return. Using the Narajo scoring system the score for this patient was probable adverse drug reaction. McEwan published a report on a Siamese cat that developed PF associated with the administration of cimetidine. In that case the P-30 histopathological findings showed changes consistent Toxic epidermal necrolysis in a dog after a with PF and also vasculitis. In conclusion this would triamcinolone injection appear to be a case of vaccine-induced PF and possible vasculitis. V. BALAZS Source of funding: University of Liverpool, Liverpool, Oftaderm, Clınica especializada en Oftalmologıa y UK. Dermatologıa Veterinaria, Santiago, Chile Conflict of interest: None declared. Corticosteroids are used for treating dogs with Stevens– Johnson syndrome/ toxic epidermal necrolysis (SJS/ TEN). In human medicine, there have been reports in which the administration of high-dose corticosteroids P-29 has induced SJS/TEN. In veterinary medicine such events have not been reported. We present a case of a Skin scrotal nodules in three dogs: 4-year-old, 4.9 kg female pug that was injected with immunophenotyping of the infiltrating cells 1 mg/kg triamcinolone for allergic responses; no other F. ALBANESE*, F. ABRAMO†, S. L. BENALI*, L. drug was administered. The day after the injection the LUCIANI‡, G. CALZAVARA§, A. PIRONE† and V. dog presented with depression, lethargy and anorexia. MIRAGLIOTTA† The skin showed macular and patchy erythema that *Veterinary Diagnostic Laboratory MyLav-La Vallonea, progressed to confluent erythema on the trunk, ven- Passirana di Rho (MI), Italy; †Department of trum, groin, face and inner pinna. On the third day, Veterinary Sciences, University of Pisa, Pisa, Italy; there were vesicles on the inner pinna and ventrum. ‡Free practitioner, Cattolica (RN), Italy; §Ambulatorio Epidermal detachment started on the trunk and began Veterinario San Giorgio, Padova, Italy to generalize showing pseudo-Nikolsky sign. The dog was sent to an intensive care unit. Complete blood Solitary and multiple skin fibro-histiocytic nodules are count and differential white blood count tests were uncommon findings in young dogs. Lesions are found within normal limits. Clinical chemistry panel revealed on face, scrotum and legs. Histologically they are the following concentrations: creatinine 2.5 mg/dL nonencapsulated confluent nodules composed of solid (normal 0.4–1.8); alanine aminotransferase (ALT) sheets of plumped and spindle-shaped cells morphologi- 113 U/L (normal 22.0–35.0); total bilirubin 12 mg/dL cally consistent with histiocytes or fibroblasts. The (normal 0.15–1.45); albumin 1.6 mg/dL (normal 2.8– immunophenotype of the cells composing the lesion has 4.0). Blood gas test indicated 4.5 mmol/L (normal <2.5) never been investigated. Because differential diagnosis lactate. Systemic arterial pressure was 60 mmHg (nor- includes a range of infectious and noninfectious mal 120–130 mmHg). Five days after the triamcinolone immunologically driven diseases, establishing the injection the dog died with disseminated intravascular immunophenotype of the infiltrating cells might be of coagulation due to systemic failure. A skin biopsy was value for therapeutic purposes. We describe three cases

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 Abstracts 67 not performed. The diagnosis was toxic epidermal 8-year-old spayed female shar-pei was presented with necrolysis triggered by an overdose of triamcinolone. two symmetrical firm plaques in the caudal part of Veterinarians should be aware of the possibility of life- both thighs, a smaller plaque and two circular ulcers threatening adverse reactions by overdoses of systemic with well-defined borders and reticulated hyperpigmen- long-lasting corticosteroids. tation in the right flank. The patient was under treat- Source of funding: Self-funded. ment with 0.5 mg/kg of prednisone every 48 h owing to Conflict of interest: None declared. recurrent bouts of fever. General physical examination was normal. Complete blood cell count, serum biochemistry, urinalysis and serological testing for Leish- mania, Anaplasma, Ehrlichia and filaria were normal or P-31 negative. A deep skin biopsy for histopathological examination and bacterial culture was performed. Bac- Canine perianal fistulas in a German shepherd terial culture was negative and the dermatopathological cross dog examination revealed a neutrophilic panniculitis and A. L. BOO vasculitis. This patient harboured one mutant allele of Pet First Veterinary Centre, Kuala Lumpur, Malaysia the mis-sense mutation of the MTBP gene. Complete clinical response was observed after three weeks admin- This is a case report of canine perianal fistulas in a 3- istration of 1 mg/kg prednisone once daily, with subse- year-8-month-old male German shepherd cross dog. He quent reduction of the dose during the following was presented for second opinion of nonhealing peri- month. Relapses were not seen during the following six anal wound. The dog was first treated by the primary months. To the best of the authors’ knowledge, this is veterinarian with metronidazole and papase for a week, the first case of SPAID reported with cutaneous neu- with no improvement seen in the perianal lesions. The trophilic panniculitis and vasculitis. reported clinical signs included perianal wound, fre- Source of funding: Self-funded. quent scooting, perianal licking, perianal pain, Conflict of interest: None declared. increased frequency of defaecation and aggressive behaviour when lifting up the tail. Physical examinations revealed swollen perianal area with fistulas and mucop- urulent discharge at the fistulas. Impression smear of the perianal lesions showed the presence of intracellular P-33 cocci and degenerated neutrophils indicating infection. A case of sterile eosinophilic pustulosis and A tentative diagnosis of canine perianal fistulas was subcorneal pustular dermatosis in a canine made based on the history, clinical signs and physical patient examination findings. Immunomodulatory therapy was † initiated with ciclosporin, 4.5 mg/kg per os once daily M. CARDOSO*, G. FREDO and G. ‡ and cefalexin, 22.7 mg/kg p.o. twice daily, to treat the CASAGRANDE bacterial infection. The perianal lesions showed marked *Dermalove Veterinary Dermatological Office, Caxias † improvement after initiation of treatment and contin- do Sul, RS, Brazil; Diagnose Pathology and Citology ‡ ued to improve when the ciclosporin regimen was mod- Laboratory, Caxias do Sul, RS, Brazil; Conforto ified and combined with ketoconazole, 5 mg/kg/day Animal Veterinary Clinic, Caxias do Sul, RS, Brazil p.o. during the subsequent follow-up. One month after The aim of this report was to demonstrate the effi- initiation of ciclosporin therapy 0.1% tacrolimus oint- ciency of ciclosporin in treating sterile eosinophilic pus- ment was introduced as a topical immunomodulatory tulosis, which is a rare idiopathic dermatosis in dogs. A agent. The patient showed continuous improvement 3-year-old, female neutered Chihuahua arrived at the and was lost to follow-up two months after the initia- veterinary dermatological service for severe itching clas- tion of treatment. sified at level 5 intensity of Hill’s Scale. The examina- Source of funding: Self-funded. tion revealed erythematous follicular and nonfollicular Conflict of interest: None declared. papules and pustules, annular erosions and epidermal collarettes in the ventral, dorsal and face regions. Antibiotic therapy was cefalexin 30 mg/kg twice daily per os and prednisolone 1 mg/kg once daily for 14 P-32 days, with improvement and then worsening when Cutaneous neutrophilic panniculitis and treatment was suspended. Allergic exclusion tests were performed using topical ectoparasiticide and dietary vasculitis associated with shar-pei management with a hypoallergenic ration, and the autoinflammatory disease – a case report patient did not tolerate suspending the corticosteroid M. CABRE*, L. ORDEIX*,†, R. GARCIA- beyond 48 h. Blood tests were performed with no FONTICOBA* and L. FERRER*,† changes; skin biopsy revealed multifocal epidermal *Fundacio Hospital Clınic Veterinari, Universitat hyperplasia, formation of subcorneal cracks with pre- Autonoma de Barcelona, Bellaterra, Spain; dominance of eosinophils. In the superficial dermis †Departament de Medicina i Cirurgia Animals, infiltrates included a moderate presence of eosinophils, Universitat Autonoma de Barcelona, Bellaterra, Spain and occasional mastocytes, neutrophils and histiocytes. The initial treatment was cetirizine 2 mg/kg twice daily, Skin lesions described in shar-pei autoinflammatory dis- without response. Treatment was changed to pred- ease (SPAID) are clinically unspecific and histopatho- nisolone 1 mg/kg once daily for two weeks. After that, logically poorly characterized. The objective of this case prednisolone 1 mg/kg every 48 h for one week, in com- report was to describe a shar-pei dog with SPAID and bination with ciclosporin at 7 mg/kg once daily. After cutaneous neutrophilic panniculitis and vasculitis. An 60 days of treatment, the lesions did not return, and

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 68 Abstracts the pruritus was classified as intensity level 2. Ciclos- P-35 porin appeared to assist in the control of sterile eosinophilic pustulosis, producing a corticosteroid-sparing Extra-auricular lesions of proliferative and effect and likely increasing the patient’s life expectancy. necrotizing externa otitis in two kittens Source of funding: Self-funded. P. PANZUTI, O. JONGH, M. MOSCA, S. VIAL and Conflict of interest: None declared. D. PIN VetAgro Sup, Marcy l’Etoile, France Proliferative and necrotizing otitis externa (PNOE) is a P-34 rare disease affecting the external ear canals and con- cave pinnae of cats. We report two cases of PNOE with Erythema multiforme as a paraneoplastic extra-auricular lesions. Case 1, an 8-month-old domes- syndrome in a dog with chronic lymphocytic tic short hair (DSH) kitten was presented with bilateral leukaemia: case report chronic otitis, suggestive of PNOE, and had unusual hyperkeratotic skin lesions along the eyelids. Case 2, a V. GMYTERCO, B. PACHECO, L. 6-month-old DSH kitten, was presented with a four KLUTHCOVSKY, F. NOGUEIRA and M. FARIAS month history of bilateral otitis, suggestive of PNOE, Postgraduate Program in Animal Science, School of Life associated with numerous arciform to nummular hyper- Sciences, Pontifıcia Universidade Catolica do Parana, keratotic skin lesions overlying erosions disseminated Curitiba, PR, Brazil over the entire body. In both cases, histopathological Erythema multiforme is an uncommon dermatopathy examination of skin biopsies from ears and extra-auri- in which the keratinocyte is the target of an immune cular lesions showed a marked orthokeratotic and response and induction of apoptosis. Herpes viruses parakeratotic hyperkeratosis and papillomatous hyper- can trigger the disease in humans, whereas in dogs it plasia of the epidermis and of the outer root sheath of has been associated with drugs, infections, idiopathic hair follicles. Numerous individually shrunken and rashes and rarely with lymphoreticular neoplasia. This hypereosinophilic keratinocytes with pyknosis were study aims to report a case of erythema multiforme observed at all levels of the epidermis and outer root triggered by chronic lymphocytic leukaemia. One 6- sheath of hair follicles. A marked lymphocytic exocyto- year-old, female shih-tzu was evaluated owing to sis was present in all levels of the epidermis along with chronic papular-crusted and intensely pruritic lesions satellitosis. Case 1 resolved spontaneously after two with multifocal distribution not responding to previous months. In Case 2, topical tacrolimus (Protopic 0.1% treatment with antibiotics and topical antiseptic ther- ointment, LEO Pharma A/S; Ballerup, Danemark) was apy. There were arciform macules, papular and erythe- applied into ears. Topical application to the dissemi- matous lesions covered by crusts on the abdomen, nated lesions was impractical so oral ciclosporin (Cycla- trunk, limbs, interdigital and periorbital areas. vance, Virbac; Carros, France; 7 mg/kg/day) was Histopathological examination revealed keratinocyte added. Clinical remission was observed after two apoptosis in all layers of the epidermis that formed months for ear lesions and other skin lesions are still necrosis. Complete blood count (CBC) revealed mild ongoing at the time of writing. To the best of the anaemia and lymphocytosis (12,496 lymphocytes/lL). authors’ knowledge, this is the first report of extra-auri- Serum biochemical results showed hyperproteinaemia cular lesions associated with PNOE in cats. due to hyperglobulinaemia. Myelogram showed ery- Source of funding: Self-funded. throid hypoplasia and an increased number of mature Conflict of interest: None declared. lymphocytes compatible with chronic lymphocytic leukaemia (CLL). Quantitative real-time PCR of blood samples indicated the prevalence of clonal population of T lymphocytes. Based on clinical and histopathologi- P-36 cal signs, the diagnosis established was paraneoplastic erythema multiforme. Treatment included chlorambucil Efficacy of dietetic food on generalized (0.2 mg/kg per os, once daily for seven days and then idiopathic ischaemic dermatopathy in three continuously at 0.1 mg/kg every 48 h) combined with miniature dachshunds prednisolone (1 mg/kg p.o. once daily). After six † K. SHIBATA* and Y. KAGAWA months of treatment, CBC normalized and skin lesions *DVMs Animal Medical Center Yokohama, Kanagawa, improved, although recurrences were common. This Japan; †North Lab, Hokkaido, Japan case report suggests that erythema multiforme, when chronic and difficult to control, may have a paraneo- Generalized idiopathic ischaemic dermatopathy (GIID) plastic origin. is a type of ischaemic dermatopathy that is adult-onset Source of funding: supported by the coordination for the and has no correlation to vaccines. Treatment of improvement of higher education personnel – Brazil ischaemic dermatopathy includes immunosuppressive or (CAPES). immunomodulatory agents and vitamin E. We hypothe- Conflict of interest: None declared. sized that Derm Defense (DD, Hill’s-Colgate (Japan) Ltd; Nibancho Chiyoda-Ku, Japan) dog food enriched with vitamin E may have potential as treatment for this condition, similar to other types of vitamin E administration. The onset with GIID on average is in 7- to 15- year-old dogs. The dogs in this study had oedematous alopecia with scale or crust on the pinnae. Other skin lesions included necrosis of the pinna, alopecia on the face, on the tail tip and over bony prominence areas.

They also had weight loss and mild myoatrophy. oclacitinib. Oedema and crusts on the pinnae improved Histopathological findings of two dogs biopsied showed within two months of treatment, at which time oclac- follicular atrophy with mild fibrosis around the hair itinib was stopped. Four months later, the skin lesions follicles. Two cases were fed DD without any drugs. In on the pinnae were resolved. The dogs regained lost one case, oedema on the pinnae decreased over time weight. These findings support DD as having efficacy and no other treatment was needed. In another case, as an adjunctive treatment in GIID cases. although crusts on the pinnae disappeared after two Source: Derm Defense was gifted by Hill’s-Colgate months of consuming DD, the dog needed concurrent (Japan). treatment to control scratching behaviour and oedema Conflict of interest: KS has lectured for Hill’s-Colgate on the pinnae. The last case was treated with DD and (Japan).

P-37 female neutered beagle presented with oral papillomas that began at six months of age after recovering from Canine cutaneous pigmented viral plaques: a distemper. The lesions were multifocal verrucous nod- clinicopathological and molecular study ules localized to the lips, gingiva and buccal mucosa. M. MAZZEI*, M. ORLANDI†, M. FORZAN*, M. Complete blood cell count and differential white blood MEI*, F. ALBANESE†, F. LEONE†,M. cell count were within normal limits. The chemistry VASCELLARI‡ and F. ABRAMO* profile only revealed a two-fold elevation of alkaline *Department of Veterinary Sciences, University of Pisa, phosphatase and a slight elevation of alanine amino- Pisa, Italy; †Veterinary Diagnostic Laboratory La transferase. Other values were within normal limits. Vallonea, Passirana di Rho (MI), Italy; Distemper PCR was negative. A biopsy was not per- ‡Histopathology Department, Istituto Zooprofilattico formed. There was no response to therapy with azithro- Sperimentale delle Venezie, Legnaro (PD), Italy mycin nor autologous papillomavirus vaccine. The dog was anaesthetized and the lesions were treated using Currently, 23 types of canine papillomavirus (CPV) cryotherapy with six freeze–thaw cycles per lesion belonging to Chi, Lambda and Tau genera have been (freeze times 6 s). One month later, the size and num- identified as responsible for oral papillomatosis, exo- ber of the papillomas had reduced significantly. A small phytic cutaneous papilloma, inverted papilloma and amount of 70% trichloroacetic acid was applied once pigmented viral plaques. Twelve types of PV, all to each lesion and allowed to dry. One month later the belonging to the Chi genus, are reported as aetiological lesions had resolved completely. Treatment success in agents of pigmented viral plaques. Our objectives were this case suggests that cryotherapy complemented with to: (i) investigate the clinical and histopathological a solution of trichloroacetic acid may be an effective characteristics of canine pigmented viral plaques; (ii) therapy for persistent oral canine papillomatosis. In this evaluate the location of viral antigen in epithelial cells case, owing to the long-lasting period of this disease, it by immunohistochemical (IHC) analysis; and (iii) iden- is unlikely that the oral papillomas could have resolved tify the CPV type using PCR. Cases were retrieved ret- without treatment. rospectively from histopathology laboratories and Source of funding: Self-funded. clinical history and signalment data were collected. Conflict of interest: None declared. IHC analysis was performed using the human PV mouse monoclonal antibody (ab-2417, AbCam; Cam- bridge, UK). Extracted DNA was amplified by PCR with generic or specific sets of primers. Sixteen cases were collected. Lesions were circular with irregular P-39 margins, flat or raised, hyperpigmented, single or multi- Ineffectiveness of marbofloxacin to treat focal and often grouped. Histologically all samples canine leproid granuloma: a case report showed a scalloped surface configuration with hyper- R. SILVA*, B. MANIER*, T. COSTA*, C. plasia, hyperkeratosis, hyperpigmentation and presence † of few koilocytes. IHC identified the viral capsid pro- BOTELHO* and J. FERNANDES *Postgraduation program, Universidade Federal Rural do tein L1 in 14/16 cases. PCR with generic and CPV-spe- † cific primers, and sequencing allowed identifying the Rio de Janeiro, Rio de Janeiro, Brazil; Department of viral genotype CPV-4, CPV-8 and CPV-9 in 2/14, 2/14 Veterinary Science, Universidade Federal Rural do Rio and 10/14 cases, respectively. In the two PCR-negative de Janeiro, Rio de Janeiro, Brazil samples, IHC scored positive. The combined diagnostic Canine leproid granuloma is caused by alcohol-acid methods used in this study confirmed the role of differ- resistant bacillary bacteria, which does not stain with ent PVs in all selected cases. Differently from other PV routine stains such as Diff-Quick. It mainly affects ani- types, CPV8 was found in young female dogs on the mals with short coats and body regions such as the face and flank. pinna with superficial vascularization and low hair den- Source of funding: University of Pisa. sity. The diagnosis should be made via histopathologi- Conflict of interest: None declared. cal evaluation using Ziehl–Neelsen stain. Immunohistochemical analysis and PCR can be used in order to identify the bacterial species involved. The treatment involves the use of antibiotics and most com- P-38 monly fluoroquinolones, doxycycline and clar- ithromycin, as monotherapy or combination. A dog Successful resolution of a case of canine oral with a nodular, alopecic, nonpruritic lesion located to papillomatosis with cryotherapy and a 70% the convex aspect of the pinna was presented at the solution of trichloroacetic acid Veterinary Hospital of the Universidade Federal Rural V. BALAZS and B. RIVERA do Rio de Janeiro. Cytological examination showed a Oftaderm, Clınica especializada en Oftalmologıa y pyogranulomatous inflammation and the presence of Dermatologıa Veterinaria, Santiago, Chile unstained filamentous structures (negative image) suggestive of Mycobacterium organisms. Histopathological Canine papillomaviruses can affect the skin and examination identified the presence of bacillary struc- mucous membranes of dogs. Most lesions regress spon- tures within macrophages, confirming the diagnosis of taneously; however, severe cases may be refractory to canine leproid granuloma. Oral marbofloxacin was ini- treatment. There is scarce published information on the tiated at a dose of 2.75 mg/kg every 24 h for 30 days. use of cryotherapy or trichloroacetic acid for treating After 35 days, the dog owner reported the appearance oral papillomatosis in dogs. A 2-year-old, 9.4 kg,

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 Abstracts 71 of new lesions; therefore, it was decided to change the P-41 medication to enrofloxacin combined with doxycycline, which led to complete resolution of the lesions in 30 Efficacy of mupirocin or chlorhexidine in the days. Despite canine leproid granuloma being consid- topical treatment of canine superficial ered a self-limiting disease, the lesions improved only pyoderma caused by resistant and after changing the treatment. Oral administration of multiresistant bacteria marbofloxacin at 2.75 mg/kg every 24 h was not effec- R. GAERTNER, V. GMYTERCO and M. FARIAS tive for the treatment of canine leproid granuloma in Postgraduate Program in Animal Science, School of Life this case. Sciences, Pontifical Catholic University of Parana Source of funding: Partially funded by the Coordenacßao~ (PUCPR), Parana, Brazil de Aperfeicßoamento de Pessoal de Nıvel Superior-Brasil (CAPES). Superficial pyoderma (SP) is a frequent cutaneous Conflict of interest: None declared. infection of dogs, usually caused by Staphylococcus pseudintermedius. Its recurrence is commonly associated with allergic, seborrhoeic or endocrine skin diseases, and the regular use of systemic antibiotics for its con- P-40 trol has been associated with meticillin-resistant (MR) and multidrug-resistant (MDR) infections. This study Isolation of strains of Sporothrix schenkii evaluated the efficacy of topical 0.2% mupirocin and complex subjected to freezing 4% chlorhexidine mousse in the treatment of SP in F. CARVALHO*,†, P. VITOR‡, M. MACHADO§ and dogs with MR and MDR strains of Staphylococcus J. FERNANDES§ intermedius group (SIG). We selected eight dogs with *College of Veterinary Medicine, Centro Universitario recurrent multifocal SP characterized by impetigo and de Valencßa, RJ, Brazil; †College of Veterinary Medicine, folliculitis, secondary to atopic dermatitis. Culture and Universidade Iguacßu, RJ, Brazil; ‡Private Veterinary antibiogram were performed in samples collected from Clinic, Veterinaria Montese, Resende, RJ, Brazil; primary lesions. Meticillin-resistant SIG was isolated in §Department of Veterinary Science, Universidade Federal five dogs and MDR SIG in three. Five dogs (two MR Rural do Rio de Janeiro, RJ, Brazil SIG and three MDR SIG) were treated topically with 0.2% mupirocin spray and three (MR SIG) were trea- Sporotrichosis is a deep mycotic infection caused by ted with 4% chlorhexidine mousse, twice daily for 21 fungi belonging to the Sporothrix schenckii complex. days. Lesion severity was assessed by a numerical lesion The zoonotic nature of sporotrichosis results in great score ranging from zero to six and the degree of pruri- public health concerns. Domestic cats are an important tus by a validated pruritus Visual Analog Scale source of fungal dissemination through their nails and (pVAS), weekly for 21 days. Response to treatment was oral cavity, and can infect humans and other animals evaluated by ANOVA and P < 0.05 considered signifi- mainly by the traumatic implant of the agent through cant. Reduction of lesion scores and degree of pruritus the skin. From the moment that an animal dies, proper was significant (P < 0.05) in both treatments, suggest- handling of the carcass represents a critical point to ing that both treatments were effective in controlling avoid the dissemination of the agent. This work aimed recurrent MR SP. Moreover, treatment with 0.2% to investigate the presence of viable and potentially mupirocin was effective in controlling MDR strains. infectious forms of the fungus in carcasses of cats diag- Source of funding: Drogavet Veterinary Handling Phar- – ° nosed with sporotrichosis kept under 18 C tempera- macy provided the aqueous spray of 0.2% mupirocin and tures. Skin and nail samples of 20 frozen carcasses were the 4% chlorhexidine mousse. collected and plated onto Mycosel (HiMedia; Mumbai, Conflict of interest: None declared. India) agar. Half of the samples were cultured in an oven at 37°C and half at room temperature (25°C). All samples were incubated for seven days and then analysed in the laboratory. Fungi belonging to the P-42 S. schenckii complex were isolated from five of 20 samples. The mycelial form of the fungus was identified in Cutaneous protothecosis caused by Prototheca four carcass samples and the yeast form in one. The wicherhamii infection in a dog study findings demonstrate that Sporothrix organisms V. GMYTERCO*, L. LUDWIG*, T. JAGIELSKI†, can still be viable in carcasses even under freezing con- ‡ M. RIBEIRO and M. FARIAS* ditions. Therefore, carcasses should be considered an *Postgraduate Program in Animal Science, School of important source of zoonotic infection and environmen- Life Sciences, Pontifical Catholic University of Parana tal contamination. (PUCPR), Parana, Brazil; †Department of Applied Source of funding: Self-funded. Microbiology, Institute of Microbiology, Faculty of Conflict of interest: None declared. Biology, University of Warsaw, Poland; ‡Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine and Animal Sciences, Universidade Estadual Paulista, Botucatu, SP, Brazil Protothecosis is an infectious disease caused by unicel- lular, achlorophyllous microalgae ubiquitous in nature. Opportunistic infections occur in humans and animals. In dogs, Prototheca zopffi is the most common pathogenic species, causing enteric, ocular and/or neurological signs. Cutaneous lesions are less frequent, often

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 72 Abstracts misdiagnosed or unnoticed, and caused by P. wicker- P-44 hamii. We report an uncommon case of protothecosis in a 2-year-old, mixed-breed dog with a history of cuta- In vitro evaluation of the bactericidal activity neous lesions of four months duration and contact with and residual effect of biocidal sprays on canine wastewater. Physical examination showed exudative hair against meticillin-resistant Staphylococcus nasolabial plaques and ulcerated, erythematous and pseudintermedius painful lesions on all foot pads associated with enlarge- A. ALVA*, D. MEZA*, C. CHUQUIZUTA*, E. ment of regional lymph nodes. Haematological and bio- † † chemical examinations were unremarkable. Histological ESCOBEDO and G. MONTOYA examination revealed a severe cutaneous inflammatory *Department of Microbiology, Universidad Cientıfica del Sur, School of Veterinary Medicine, Lima, Peru; reaction, with numerous periodic acid Schiff-stained † round or oval encapsulated structures, compatible with Department of Dermatology, Universidad Cientıfica del Prototheca species. Microbiological culture of cutaneous Sur, School of Veterinary Medicine, Lima, Peru exudate on Sabouraud agar kept in aerobic conditions, There is an urgent need to investigate alternative and at 37°C, revealed white colonies after 48 h. Phenotypic- effective topical treatments against skin infectious dis- based identification using a newly described PCR- eases. The study aim was to assess the efficacy and restriction enzyme analysis (PCR-REA) based on the residual effect of the following biocidal sprays: cytB gene, confirmed the organism identity as Pro- chlorhexidine 2% (Durvet, Durvet), sodium hypochlo- totecha wicherhamii. Oral itraconazole was initiated at rite 0.01% (Clorox, Clorox Peru SA), hypochlorous 10 mg/kg once daily and partial improvement was acid 0.65% (Ecaderm, Eca Control SAC) and N-acetyl- observed after eight weeks of treatment, with total reso- cysteine 0.04% (Fluimucil, Zambon SpA). Sodium lution after six months. However, recurrence occurred chloride 0.9% was used as negative control. Each spray after two months of therapy discontinuation and the was applied once on the flank of 30 healthy dogs cover- treatment was changed to oral terbinafine at 20 mg/kg ing an area of 25 cm2 and 2 cm distant from each twice daily, unsuccessfully. Itraconazole was re-started other. Dog hairs (0.02 g) were collected daily for the with partial improvement of lesions at this time. We first four days post-treatment and placed equidistant highlight the difficulty in treating P. wicherhamii infec- from each other onto a Mueller–Hinton agar plate tion and the public health concern given that cutaneous streaked with meticillin-resistant Staphylococcus pseud- and systemic P. wicherhamii infection has been reported intermedius (MRSP) isolated from a dog with pyo- in humans. derma. The size of the inhibition halo was measured Source of funding: Self-funded. every 24 h. Chlorhexidine 2% had a residual effect of Conflict of interest: None declared. three days, forming an inhibition halo of 2.4 Æ 0.5 cm on day (D)1, 1.7 Æ 0.5 cm on D2 and 1.0 Æ 0.6 cm on D3. The other sprays showed an inhibition halo; however, focal areas of bacterial growth surrounding the P-43 hairs were observed after D1. On D1, the halo for the Æ Antifungal susceptibility of ravuconazole sodium hypochlorite spray was 1.6 0.5 cm, for hypochlorous acid 1.6 Æ 0.5 cm and for N-acetylcys- against dermatophyte isolates from animals teine 1.5 Æ 0.4 cm. In conclusion, chlorhexidine 2% R. KANO, H. FURUTA and H. KAMATA spray was more effective in inhibiting MRSP growth College of Bioresource Sciences, Nihon University, in vitro than the other products and had a longer resid- Fujisawa, Japan ual effect. Dermatophytosis is a common cutaneous mycosis in Source of funding: Self-funded. small animals and frequently causes human infection. Conflict of interest: None declared. Ravuconazole (RVCZ) is a newly available (approved for human use in Japan in 2018) broad-spectrum antifungal azole that exhibits excellent activity against dermatophytes. To the best of the authors’ knowledge, this P-45 is the first study to investigate the in vitro susceptibility Association of prior antibiotic exposure to of RVCZ against dermatophyte isolates from animals isolation of meticillin-resistant Staphylococcus and a terbinafine (TERB)-resistant strain of Microspo- rum canis. The Clinical and Laboratory Standards pseudintermedius from dogs with skin and ear Institute (CLSI) M38-A2 test was used in the study. infections The minimal inhibitory concentrations (MICs) of K. K. MWACALIMBA, V. FADOK, S. CAMPBELL RVCZ for 17 clinical isolates of M. canis ranged from and D. AMODIE 0.03125 to 0.5 mg/L; for three clinical isolates of Zoetis LLC, Parsippany, NJ, USA M. gypseum from <0.03125 to 0.03125 mg/L; for one This study examined the association between antibiotic clinical isolate of Trichophyton benhamiae <0.03125 mg/ exposure and clinical isolation of meticillin-resistant L; and for nine clinical isolates of T. mentagrophytes Staphylococcus pseudintermedius (MRSP) from dogs <0.03125 mg/L. The MICs for the TERB-resistant using the electronic database of a hybrid practice M. canis strain (47C) were 0.03125 mg/L for RVCZ, located in the United States, from August 2011 to 0.025 mg/L for itraconazole (ITCZ) and >32 mg/L for March 2018. Antibiotics included fluoroquinolones, TERB. The results of the present study indicate that cefalexin, cefovecin, cefpodoxime, clindamycin or RVCZ might be more effective than ITCZ and TERB ‘other’, used within 180 days before culture. Compar- against clinical isolates of dermatophytes. isons were based on proportion analysis (SAS 9.4; Source of funding: Self-funded. Cary, NC, USA) using a two-sided Student’s t-test Conflict of interest: None declared. (P ≤ 0.05). Five hundred and seventy-eight

S. pseudintermedius clinical isolates (20, abscess; two, sensitivity of M. pachydermatis to ketoconazole and cyst; 69, ear; 487, skin) from 503 dogs were reviewed; posaconazole. 50% of the isolates (289) were MRSP. Clinical isolates Source of funding: Self-funded. from skin had the highest rate of MRSP (51.5%, 251/ Conflict of interest: None declared. 487). Dogs received between none to six antibiotic courses in the 180 days before culture. Of the 578 isolates, 33% were from dogs with no history of antibiotic exposure, while 35% had received antibiotics once. P-47 Fifty dogs had more than one culture: 21 were previously MRSP but not MRSP from next isolate; 14 were Control of dermatophytosis in guinea pigs: previously non-MRSP and MRSP on a subsequent cul- comparison of procedures in 83 cases ture. Dogs with at least one exposure to antibiotics had P. BOURDEAU, A. LE FRIEC, C. DUMONT, M. 44–58% MRSP isolates. Forty-seven percent (140/300) MUKOKO, F. ALEXANDRE and C. ROUX of isolates from dogs that received one or two courses Laboratory LDPM LabOniris, Department of Clinical of antibiotics were MRSP (P = 0.1025) and 60% (50/ Sciences, Unit DPM, Veterinary School (Oniris), 83) of isolates from dogs that received three or four University of Nantes, France courses of antibiotics were MRSP (P = 0.0083). These Although guinea pigs have been widely used in experi- results support prior literature that previous antimicro- mental models for studying antifungals, there is virtu- bial therapy is an important risk factor for the isolation ally no comparative information on methods for of MRSP. control of dermatophytosis in practice. The information Source of funding: Zoetis. in this study was obtained from the authors’ labora- Conflict of interest: KM, VF, SC and DA are employees tory. The protocols used in practice and their efficacy of Zoetis, Inc. (post-treatment negative cultures) were compared using chi-squared tests. From a series of 83 cases, enilconazole 0.2% was used in 83% of animals; every four days during six weeks in 66% and as the only therapy in P-46 39%. Itraconazole (5 mg/kg/day) was used in 90% of Effect of repeated use of suboptimal the 83 cases; in an alternate week therapy in 89%, three concentration of enilconazole on sensitivity of times in 91% and with 60% (6/10) efficacy when used Malassezia pachydermatis to ketoconazole and alone (a prolonged daily treatment cured 3/3 guinea pigs). Griseofulvin was used in 10% of the 83 cases at posaconazole 20–40 mg/kg (usually 25) daily over four weeks in P. BOURDEAU, C. THIBAULT, C. DUMONT and 60%. The tolerance was always excellent (occasional C. ROUX difficulties in administering enilconazole or griseoful- Laboratory LDPM LabOniris, Department of Clinical vin). Combined systemic and topical treatment (n = 36) Sciences, Unit DPM, Veterinary School (Oniris), was the most effective [86% versus 81% (only topical) University of Nantes, France and 69% (only systemic)] (P=0.494). All long-haired The frequent use of antifungal medications in dogs can animals (4/4) were cured when clipped (versus 5/8 result in changes in the sensitivity of Malassezia organ- unclipped). Both disinfection of environment (69% cure = isms to these drugs. This study was conducted to assess versus 31% without disinfection; P 0.277) and treat- the effects of suboptimal use of enilconazole on the sen- ment of conspecifics (86% cure versus zero cure if not sitivity of Malassezia organisms to ketoconazole and treated) helped. From these results we propose the posaconazole. Twenty healthy beagles with bilateral combination of enilconazole and itraconazole for six aural populations of M. pachydermatis were included. weeks with clipping of long-haired animals, treatment On day (D)0 and D5 one ear of each dog received of conspecifics and disinfection of the environment as 0.5 mL of a sub-therapeutic concentration of enilcona- the most effective management of dermatophytosis in zole (0.05%). Evolution of sensitivity of the yeast guinea pigs. organisms to two azoles (ketoconazole and posacona- Source of funding: Self-funded. zole) was evaluated before treatment on D0 and D10 Conflict of interest: None declared. using ETEST (BioMerieux; Marcy-l’Etoile, France) strips. Minimum inhibitory concentration (MIC; µg/ mL) ranges in untreated ears included: ketoconazole D0, 0.125–0.25 µg/mL (median 0.19) and D10, 0.125– P-48 0.38 µg/mL (0.19); posaconazole D0, 0.0125–0.25 µg/ Application of the ETEST technique to mL (0.125) and D10, 0.047–0.38 µg/mL (0.125). Sensi- evaluate the sensitivity of Malassezia tivity of Malassezia to ketoconazole in treated ears was 0.094–0.25 µg/mL (0.19) before treatment and 0.125– pachydermatis to ketoconazole, posaconazole 0.25 µg/mL (0.19) on D10. Likewise the sensitivity to and itraconazole in normal dogs posaconazole was 0.0125–0.25 µg/mL (0.125) before P. BOURDEAU, C. THIBAULT, F. HUBERT and F. treatment and 0.047–0.25 µg/mL (0.125) on D10. The ALEXANDRE MIC remained unchanged post-treatment in 16/20 ears Laboratory LDPM LabOniris, Department of Clinical for ketoconazole and 6/20 for posaconazole; it was Sciences, Unit DPM, Veterinary School (Oniris), higher in 2/20 ears for ketoconazole and 7/20 for University of Nantes, France posaconazole and equally lower for each antifungal. In Little is known about the sensitivity to antifungal medi- this study, the repeated application of suboptimal doses cations of resident populations of Malassezia pachyder- of enilconazole did not induce any changes in the matis in dogs. The objective of this study was to

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 74 Abstracts evaluate the activity of ketoconazole, posaconazole and predictive value (98%), independent of the fungal spe- itraconazole in ear canal populations of this yeast. cies (P = 0.843). When positive, trichoscopy was indica- Malassezia pachydermatis was obtained from swabbing tive of severe infection (92% cfu > 50)(P= 2.001e-7). the ear canals of 20 healthy adult beagles. Isolated These results indicate that fungal culture is rapid, speci- colonies were used to produce suspensions (2.5 McFar- fic and sensitive for the diagnosis of dermatophytosis land) that were swabbed onto Sabouraud’s-Tween 20 and essential to evaluate mycological cure. agar. ETEST (BioMerieux; Marcy-l’Etoile, France) Source of funding: Self-funded. strips of ketoconazole, posaconazole or itraconazole Conflict of interest: None declared. then were applied to the agar. After incubation (32°C for 48 h) the minimal inhibitory concentration (MIC; µg/mL) were read directly from the strips. The MIC ranges included ketoconazole 0.094–0.25 µg/mL (me- P-50 dian 0.19), posaconazole 0.0125–0.25 µg/mL (0.125) and itraconazole 0.008–0.047 µg/mL (0.032). Left and Cowpox virus dermatitis in a cat: a case report right aural yeast populations expressed identical MICs I. OTROCKA-DOMAGAŁA*, K. PAZDZIOR- for ketoconazole and posaconazole in 9/20 dogs and CZAPULA*, M. MIKIEWICZ*, M. GESEK*, W. itraconazole in 2/7 dogs. Bilateral sensitivity profiles KRASINSKA-PRZEWORSKA † and B. were identical for posaconazole and ketoconazole in 5/ TWORKOWSKI† 20 dogs and for all three antifungals tested in 1/7 dogs. *Department of Pathological Anatomy, Faculty of There was no predictability of MIC values between Veterinary Medicine, University of Warmia and Mazury ketoconazole and posaconazole (i.e. susceptibility to in Olsztyn, Poland; †Veterinary Clinic in Olsztyn, one medication neither correlated with nor implied sus- Poland ceptibility to the other). ETEST was shown to be rapid Cowpox virus (distributed throughout Europe and and easy to evaluate sensitivities of M. pachydermatis Asia) sporadically infects outdoor hunting cats because to antifungal medications in dogs. Malassezia popula- wild rodents are the reservoir of this pathogen. The tions in ear canals express a wide range of in vitro sen- infection is manifested by skin lesions located predomi- sitivity profiles including differences between affected nantly on the head, neck and forelimbs, and some indi- ears of the same animal in a high proportion of dogs. viduals develop fatal necrotizing pneumonia. We report Source of funding: Self-funded. a case of cowpox virus infection in a cat with atypical Conflict of interest: None declared. distribution of the cutaneous lesions. A 7-year-old, neutered male European short hair, outdoor cat presented with fever (40°C), severe erythema with crusting and ulceration of the skin in the abdominal area. The cat P-49 received antibiotics and glucocorticoids with subsequent Diagnosis of dermatophytosis in guinea pigs: progression of the skin lesions. Therefore, multiple full- comparative results of fungal culture and thickness skin biopsies were sampled for histopathological evaluation, which revealed acanthosis, severe epider- trichoscopy from 400 cases mal spongiosis and ulceration. Large, irregular, P. BOURDEAU, A. LE FRIEC, F. HUBERT, M. polygonal eosinophilic inclusion bodies were seen MUKOKO, F. ALEXANDRE and C. ROUX within the cytoplasm of keratinocytes in both the epi- Laboratory LDPM LabOniris, Department of Clinical dermis and hair follicles. Massive, multifocal to coalesc- Sciences, Unit DPM, Veterinary School (Oniris), ing infiltrations of macrophages, eosinophils, University of Nantes, France neutrophils and plasma cells were observed in the The objective of this study was to describe information subepidermal and perifollicular areas, extending to the obtained from fungal culture or trichoscopy in cases of interstitial dermis. These microscopic changes were dermatophytosis in guinea pigs. Samples (carpet compatible with cowpox dermatitis. Cowpox virus method) sent to the authors’ laboratory were cultured (Orthopoxvirus) infection was confirmed by PCR test- under laboratory conditions for fungal identification ing of the epidermal crusts sampled after initial and growth quantification (colony forming units). The histopathological diagnosis. Within a few weeks, skin results were compared to trichoscopy using the chi- lesions started to heal with nearly complete recovery squared test. In the period 2016–2017, 400/740 (54%) during the following three months. Owing to the zoo- samples were positive for dermatophytes. When com- notic potential of cowpox virus, this infection should bined with 261 positive samples from 2014–2015, the always be regarded as a differential diagnosis when a frequencies of 661 dermatophyte species included: Tri- hunting cat presents with ulcerative skin lesions located chophyton porcellae 82.8%, T. mentagrophytes 14.5%, anywhere on the body. Microsporum canis 1.4% and two concurrent species Source of funding: Self-funded. 1.4%. The respective times for identification (days Æ Conflict of interest: None declared. SD) were 6.2 Æ 1.61, 5.7 Æ 1.61, 7.1 Æ 1.97 and 6.9 Æ 1.45 (P = 0.108–0.49). The cfu total was significantly higher for T. porcellae and T. mentagrophytes (56% and 61% cfu > 50) than M. canis (67% cfu = 1) (P = 0.0078). Post-treatment evaluation indicated that 21.2% T. porcellae and 9.5% T. mentagrophytes cases remained infected, mainly when initial cfu > 50 (27%) (P = 0.599). Trichoscopy (n = 584) had a limited sensitivity (17%) and negative predictive value (43%) yet was highly specific (99%) with a high positive

P-51 pyoderma outbreaks in the previous year. Allergy medications were permitted if the dogs were treated for at Tongue lesions as a clinical manifestation of least three months previously; allergen immunotherapy canine leishmaniosis in an endemic area was permitted if started one year previously. Antibiotics R. LIMA* and C. SOUZA† and topical therapy were permitted as needed. Dogs *Clinica Veterinaria Canis & Catus, RN, Brazil; admitted into the study received either topical PYOspot †Department of Veterinary Clinical Medicine, University or a topical placebo applied once weekly. Owners and of Illinois, IL, USA dermatologists were blinded, and the dogs were randomly assigned to either arm. Each dog was evaluated Canine leishmaniosis is a vector-borne protozoan dis- by a dermatologist at least four times over the follow- ease endemic to >70 countries. The disease can have ing year: after first visit (V1) at the start, V2 at approx- various clinical presentations. This report describes four imately three to four weeks to ensure pyoderma cases of canine leishmaniosis presenting with tongue resolution, V3 at six months or relapse of pyoderma lesions as the main clinical sign. The dogs were exam- and V4 at 12 months. Dogs were assessed by physical ined at a veterinary clinic in an area endemic for Leish- examination and cytological evaluation. Twenty-eight mania infantum in North-eastern Brazil. The age of the dogs from multiple centres were enrolled; two withdrew dogs at presentation ranged from four to nine years before V4. Fourteen dogs were treated with PYOspot and they were of four breeds: boxer, Rottweiler, minia- and 12 with placebo. Both groups had fewer episodes ture pinscher and pit bull. All dogs presented with ton- of pyoderma after one year (PYOspot 5.0 Æ 1.69 to gue lesions characterized by ulcers (two), coalescing 1.07 Æ 1.3; placebo 5.3 Æ 1.49 to 2.67 Æ 1.43); how- papules and plaques (two), loss of papillae appearance ever, the difference between the groups at year end was on the dorsal and lateral aspects of the tongue (two), significant (P = 0.008). We conclude that the regular erythema and papules on the ventral tongue surface use of PYOspot, in addition to treatment for the under- (one), and irregular tongue margins (one). One dog lying condition, can reduce pyoderma relapses. showed bilateral periocular ulcers and scaling with lym- Source of funding: PYOspot and placebo were provided phadenomegaly, in addition to tongue lesions. Another by Dermoscent Laboratorie. dog was previously diagnosed with leishmaniosis and Conflict of interest: None declared. had cutaneous lesions that resolved with initial therapy. Maintenance therapy with allopurinol and domperidone was administered for two years before the occurrence of tongue lesions. Cytological findings from P-53 tongue lesions of all four dogs showed amastigotes inside macrophages, cytoplasmic vacuoles and extracel- Necrotizing soft tissue infection: A case report lular amastigotes. Two dogs had a negative serological A. VILLARREAL*, D. CAMPOS†, D. BORGES† and rapid test (immunochromatography) result before the J. FERNANDES† cytological confirmation of leishmaniosis. Two dogs *Pet Skin Care Services Panama, Panama, Panama; were euthanized after the diagnosis of leishmaniosis. †Department of Veterinary Science at Universidade The remaining two dogs were treated with a combina- Federal Rural de Rio de Janeiro, RJ, Brazil tion of miltefosine, allopurinol, and domperidone. Ton- Necrotizing soft tissue infections (NSTI) are severe gue lesions resolved after 30 days of therapy. Tongue infections of the skin, subcutaneous tissue, superficial lesions should be considered as a clinical manifestation fascia, and occasionally, deep fascia, muscle and adi- of canine leishmaniosis. pose tissue. They can lead to systemic toxicity, septi- Source of funding: Self-funded. caemia and high mortality rates, if not treated in time. Conflict of interest: None declared. In this report, we describe a case of NSTI caused by a bite, in a 4-year-old female beagle. The patient presented with small puncture wounds on the sacral region and hind legs. These areas were associated with P-52 oedema, erythema and loss of sensitivity. An extensive Topical application of a proprietary blend of area of crepitation at the base of the tail also was essential oils and plant extracts is associated noted. One of the most relevant clinical signs was intense pain during examination, not correlated with with fewer relapses of pyoderma the small wounds observed. After surgical debridement, V. A. FADOK*, F. SECKERDIECK†,E. a large zone of deep necrotic tissue was observed in the BENSIGNOR‡, C. NOLI§, A. OLIVEIRA¶ and R. S. lumbosacral region, as well as purulent exudate and MUELLER† devitalized tissue. Therapy with ceftriaxone, metronida- *Zoetis, Parsippany, NJ, USA; †Ludwig Maximillian zole and tramadol chlorhydrate was initiated. In addi- University, Munich, Germany; ‡Clinique Veterinaire, tion, thorough cleaning using warm pressurized saline Paris, France; §Servizi Dermatologici Veterinari, solution and 0.01% iodopovidone solution two times a Peveragno, Italy; ¶University Lusofona de Humanidades day, as well as compression bandaging were established. e Tecnologias, Lisboa, Portugal A silver sulfadiazine paste was applied to the wounds Dogs with cutaneous disorders can develop recurrent until granulation tissue was observed, approximately 20 staphylococcal pyoderma. Addressing the underlying days after the initial debridement. The patient had a cause is necessary yet not always sufficient to control good recovery, with optimal healing and cicatrization recurrences. The purpose of this study was to determine of the involved areas. Recognizing signs of a possible the effects of weekly use of topical PYOspot (Der- NSTI can lead to an early diagnosis and treatment, moscent Laboratorie; Castres, France) on pyoderma which are fundamental factors for a good prognosis. relapse. Dogs admitted to the study had at least four Source of funding: Self-funded. Conflict of interest: None declared.

P-54 P-55 The efficacy of wipes containing 0.1% Antimicrobial resistance gene and multidrug chlorhexidine gluconate and EDTA for canine efflux pumps against antiseptics for superficial pyoderma: a pilot study staphylococci isolated from canine superficial † pyoderma in Japan Y. TAKAHASHI*, K. MORI , Y. OSHIMA- TERADA† and N. MURAYAMA† N. MURAYAMA*, Y. OSHIMA-TERADA*, K. † † *Minamino Animal Hospital, Tokyo, Japan; MORI*, H. NAKAMINAMI , T. SAITO and N. † †Dermatology Services for Dogs and Cats, Tokyo, Japan NOGUCHI *Dermatology Services for Dogs and Cats, Tokyo, Therapy with chlorhexidine-based shampoos can be † efficacious in canine superficial pyoderma yet the sham- Japan; Tokyo University of Pharmacy and Life pooing process is often laborious especially in large Sciences, Tokyo, Japan dogs. The purpose of this study was to assess the effi- Meticillin-resistant staphylococci in canine pyoderma is cacy of wipes containing 0.1% chlorhexidine gluconate a serious problem. The aim of this study was to assess and EDTA in dogs with superficial pyoderma. The mecA gene, antimicrobial resistance genes and mul- study included seven dogs from three months to 11 tidrug efflux pump genes of staphylococci from canine years of age. These dogs were diagnosed with superfi- superficial pyoderma in Japan. Bacterial isolates from cial pyoderma based on typical clinical signs and skin 136 dogs were evaluated from May 2018 to April 2019. cytological results. The owners gently wiped the lesions Staphylococci, mecA gene, antimicrobial resistance on the ventral abdomen once daily for three weeks. genes including ermA, ermB, ermC, msr, inuA, mphC, The following subjective evaluation was performed at aacA/aphD, tetM, tetK and fosB, and multidrug efflux treatment end: excellent (complete resolution), good pump genes including qacA/B and smr were identified (some improvement), fair (no improvement) and poor by PCR. Of the staphylococci, 72% were Staphylococ- (exacerbation). Erythema, papules/pustules, scales/ cus pseudintermedius, 25% S. schleiferi subsp. coagulans crusts and pruritus were scored 0–3 by the investigators and 3% unidentified species, and 47% of all staphylo- before and at treatment end (0, none; 1, occasional or cocci were mecA-positive. The staphylococci positive mild; 2, frequent or moderate; 3, severe). Cutaneous for ermA, ermB, ermC, msr, inuA, mphC, aacA/aphD, pH and transepidermal water loss (TEWL) were mea- tetM, tetK and fosB were: all staphylococci – 0%, sured before and after the trial. These data were non- 46.3%, 0.7%, 0%, 0%, 0%, 50.7%, 46.3%, 1.5% and categorical, thus the Wilcoxon signed rank test was 70.0%, respectively; MecA-negative staphylococci –0%, used to analyse differences. The subjective evaluation 22.2%, 0%, 0%, 0%, 0%, 27.8%, 33.3, 2.8% and showed five excellent, one good and one fair response. 62.5%, respectively; and MecA-positive staphylococci – Three, five, six and five dogs showed a reduction in 0%, 74.6%, 1.6%, 0%, 0%, 0%, 76.6%, 60.9%, 0% scores for erythema, papules/pustules, scales/crusts and and 78.1%, respectively. Multidrug efflux pump genes pruritus, respectively. Erythema and pruritus improved including qacA/B and smr were in 0.7% and 2.2% (re- significantly (P < 0.01). Skin pH (7.81 before, 7.46 spectively) of all isolates. MecA-positive staphylococci after: P < 0.01) and TEWL (3.13 before, 3.92 after: were positive for qacA/B and smr in 1.6% and 1.6%, P < 0.01) changed significantly. No adverse effects were respectively. MecA-negative staphylococci was positive noted in any of the dogs. This study suggest that once for smr in 2.8%. This study indicated that mecA-posi- daily 0.1% chlorhexidine gluconate and EDTA wipes tive staphylococci had many antimicrobial resistance can improve canine superficial pyoderma. genes, and a few staphylococci had multidrug efflux Source of funding: Self-founded. pump genes. Conflict of interest: None declared. Source of funding: Self-funded. Conflict of interest: None declared.

P-56 bacterial overgrowth (no inflammatory cell) (BOG; 43 ears, 41.4%), suppurative (>90% neutrophils) (Pyo; 55 Relationship between cytological, ears, 52.9%), pyogranulomatous (>50% neutrophils bacterioscopic and clinical features in 64 cases and macrophages) (PyoG; six ears, 5.8%). Chi-squared of chronic bacterial otitis externa in dogs or Fisher’s exact tests were used (P < 0.05). No correla- V. BRUET, T. BREMENT, M. RIVALAIN and P. tion was observed between the cytological pattern and BOURDEAU chronicity (more than two months: 76% BOG, 85% Dermatology, Parasitology, Mycology Service, ONIRIS, Pyo, 100% PyoG), pain, OTIS-3 or the presence of Nantes, France Malassezia. There was a statistical correlation between cytological pattern and primary factors (BOG observed The aim of this study was to characterize the relation- in 89.3% of allergic otitis) (P=0.02); number of pre- ships between bacterioscopic, clinical and cytological vious antibacterial treatment: 0 (58% BOG, 31% Pyo, features of chronic bacterial otitis in dogs. Sixty-four 17% PyoG) versus 1, 2 or 3 (P=4.6 9 10-2); clinical dogs (104 ears) with chronic bacterial otitis externa type of otitis [BOG with ErCer (61.9%), Pyo and PyoG (positive bacterioscopy confirmed by bacteriological with Sup (59.2% and 66.7%, respectively; P=5 9 10- evaluation) were included. Information on chronicity, 3];. There also was a statistical correlation with bacteria primary factors, previous topical antibacterial treat- isolated: Staphyloccocus and Corynebacterium with ments and pain was collected. Clinical scoring (OTIS-3) BOG (53.8% and 50%, respectively), Pseudomonas with and type (Er, erythematous; Cer, ceruminous; ErCer, Pyo/PyoG (80%) (P=0.04); abundance of Malassezia erythemato-ceruminous; Sup, suppurative) was [moderate, high level with BOG (84.6%); recorded. Cytological pattern [bacterial overgrowth P=2.4 9 10-3]. These results highlight the importance (BOG), suppurative, pyogranulomatous], presence of of cytological inflammatory patterns to describe otitis Malassezia and bacterioscopic results (C, cocci; R, and possible relationships with bacterial causal agents. rods) also were collected. Chi-squared or Fisher’s exact Source of funding: Self-funded. < tests were used (P 0.05). Bacterial population (mor- Conflict of interest: None declared. phological) results were: majority C, 40/104 ears (38.4%); majority R, 22/104 (21.2%); or equivalent C: R ratio (B), 42/104 (40.4%). No correlation was observed between the bacterioscopic results and pri- P-58 mary factors, pain, OTIS-3 or presence of Malassezia. A trend was observed with chronicity: more than one Bacterial chronic otitis: clinical and cytological month in 35% C, 40% B and 57% R. Statistical corre- data in 64 dogs lations were observed between the bacterial morphology V. BRUET, T. BREMENT, M. RIVALAIN and P. and the number of previous antibacterial treatments [0 BOURDEAU (55% C, 27% R) versus 1, 2 or 3 (45% C, 63%R) Dermatology, Parasitology, Mycology Service, ONIRIS, (P=0.03)], the type of otitis (Er, Cer, ErCer 15% R, Nantes, France Sup 13% C) (P=9.10-5) and the type of reaction (BOG, 53% C, pyogranulomatous 17% C) The aim of this study was to improve knowledge on (P=8.9 9 10-3). Morphology of bacteria is the first bacterial chronic otitis. For one year, dogs with chronic element for identification and this study brings new (more than two weeks) bacterial externa (CBO) were information on possible links with clinical and cytologi- prospectively analysed. Sixty-four dogs were included cal significance. (104 ears). The mean age was 6.5 years with a distribu- Source of funding: Virbac Corporate, France. tion of age older than the overall canine population of Conflict of interest: None declared. ONIRIS hospital. There was no sex predisposition. Seven breeds represented 55% of cases: French bulldog, cocker spaniel, Labrador retriever, bichon frise, Ger- man shepherd dog, beagle and West Highland white P-57 terrier. Except for Labrador retriever and beagle, all breeds were statistically over-represented compared to Bacterial chronic otitis: correlation between the hospital population. The duration was over two cytological inflammatory patterns, clinical months in >70% and the number of topical bacterial aspect and bacteriology in 64 dogs treatments before consultation 1.6. The suspected predisposing factors were hypertrichosis (9%), physiologi- V. BRUET, T. BREMENT, M. RIVALAIN and P. cal glandular hyperplasia (cocker spaniel) 9%, BOURDEAU swimming (3%) and abnormal ear conformation Dermatology, Parasitology, Mycology Service, ONIRIS, (1.5%). Primary factors were identified in 61% of cases: Nantes, France hypersensitivity (48%), foreign body (4.8%), nodule One hundred and four ears (of 64 dogs) with chronic (4.8%) and endocrinopathy (2.9%). Perpetuating fac- (minimum two weeks) bacterial otitis externa (bacteria tors were stenosis (40%) and induration (35%). In observed then identified by bacteriological evaluation) CBO, the otitis was erythematous in 7%, ceruminous were included in this study. Primary factors, chronicity, in 7%, cerumino-erythematous in 40% and suppurated previous topical antibacterial treatments, pain, clinical 46%. Cytologically, bacterial overgrowth was observed scoring (OTIS-3) and classification (Er, erythematous; in 41%, suppurative in 53% and pyogranulomatous in Cer, ceruminous; ErCer, erythemato-ceruminous; Sup, 6%. Bacteria were mainly cocci in 38%, mainly rods in suppurative) were collected. Otitis were classified by 21% and equivalent C:R ratio in 41%. Malassezia inflammatory pattern at cytological evaluation:

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 78 Abstracts often were present (>50%) as both cocci and rods (no antibiotics to control the underlying infection, in con- statistical difference) in low concentration (>75% of junction with ear cleansers to remove waxy debris and cases). This study brings new data about clinical and exudate that may encourage further microbial growth. cytological features in bacterial chronic otitis in dogs. We conducted studies to determine the antimicrobial Source of funding: Virbac Corporate, France. properties of the ear cleanser, Epiotic SIS (Virbac; Car- Conflict of interest: None declared. ros, France) against clinical isolates from Australian dogs, and whether Epiotic SIS could enhance the antimicrobial effect of Easotic (miconazole nitrate and gentamicin sulfate, Virbac), an antibiotic treatment for P-59 OE. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time–kill Lagochilascaris as a cause of unilateral kinetic assays were performed with Epiotic SIS and suppurative otitis in two cats: a case report Easotic against multiple isolates of meticillin-resistant M. CARDOSO*, F. CAGOL†, K. MATTE† and L. Staphylococcus pseudintermedius (MRSP), meticillin-sus- ANTONIO† ceptible S. pseudintermedius (MSSP), Pseudomonas *Dermalove Veterinary Dermatology Office, Caxias do aeruginosa, Proteus mirabilis, Escherichia coli and Sul, RS, Brazil; †Sao~ Francisco Veterinary Center, Malassezia pachydermatis. Epiotic SIS inhibited micro- Bento Goncßalves, RS, Brazil bial growth in all isolates (MIC 0.0156–0.125); this effect was greatest against Malassezia, MSSP and Otitis is characterized by an acute or chronic multifac- MRSP. Epiotic SIS exhibited microbicidal activity torial inflammatory disease of the auditory system. against all isolates (MBC 0.0312–1.0). Epiotic SIS killed Lagochilascariasis is a zoonosis limited to the American Pseudomonas within 4 h of exposure, and all other iso- continent caused by a nematode helminth of the Ascari- lates except for Proteus within 24 h. When Epiotic SIS didae family of the genus Lagochilascaris and is an and Easotic were tested in combination, an additive to uncommonly reported parasite in mammals. The infec- synergistic interaction was observed against MSSP and tion occurs through the ingestion of raw meat from MRSP isolates, while an additive interaction was seen hosts containing the larvae. The parasite infects subcu- against Malassezia. Epiotic SIS exhibited antimicrobial taneous regions, usually in the cervical region and sur- properties against clinical isolates of bacteria and yeast rounding tissues of the face. The aim of this report is representing the most common causes of canine OE, to provide information on the occurrence of lagochilas- including MRSP. Epiotic SIS was shown to enhance cariasis as cause of otitis media in felines and on the the effect of Easotic against common aetiological relevance of otoendoscopy as an efficient diagnostic agents of OE isolated from Australian dogs. method. In the city of Bento Goncßalves (RS, Brazil), Source of funding: Virbac, Australia. one intact 8-year-old female and an intact 1-year-old Conflicts of interest: WYC received salary from male cat, both of undefined breed, were presented with ACARE. DJT has received research funding from Luoda suppurative right unilateral otitis resulting in a putrid Pharma, Neoculi, Zoetis, Bayer, Boehringer Ingelheim, smell and pain. In the female, an otoendoscopic exam Virbac, and Elanco. revealed a polypoid, multilobulated, elongated tissue that extended from the external ear towards the middle ear. After traction removal of the polyp, cylindrical fusiform worms of a milky-white colour were observed. Forty nematode parasites of the genus Lagochilascaris P-61 were found in tissue from the female patient and four Resolution of antibiotic resistant bacteria seen were isolated from the male patient. Otoendoscopy, in in canine chronic otitis externa cases these cases, was fundamental in diagnosis, treatment and prognosis of both patients. M. FUJIMURA Source of funding: Self-funded. Fujimura Animal Allergy Hospital, Osaka, Japan Conflict of interest: None declared. Otitis is a frequent complication of repeat-occurrence allergic disorders in canines and steroids and/or antibiotics are commonly used treatment options. Because of such remedies, microbes are becoming more drug-resis- P-60 tant, and the frequency with which stronger drugs are being used is increasing. A study was performed look- In vitro antimicrobial activity of Epiotic SIS ing at the effects of an alternative, ZYMOX Otic, an and Easotic against pathogens associated with enzyme-based ear solution containing three natural canine otitis externa in Australia enzymes (lactoperoxidase, lactoferrin and lysozyme) on W. Y. CHAN*,†, A. G. KNOWLES‡ and D. J. antimicrobial activity and effectiveness in biofilm eradi- TROTT* cation. Presented are two of five clinical cases of dogs *Australian Centre for Antimicrobial Resistance with chronic dermatitis and subsequent otitis externa Ecology, School of Animal and Veterinary Sciences, The (OE). Each case presented with allergic skin disorders University of Adelaide, South Australia, Australia; and concurrent ear infections that were difficult to †Faculty of Veterinary Medicine, Universiti Putra treat. These two case studies evaluated ear conditions Malaysia, Serdang, Selangor, Malaysia; ‡Virbac pre-application and post-application of ZYMOX Otic – (Australia) Pty Ltd, Milperra, New South Wales, solution. Using a numerical scale of 0 3 (3, severe), Australia redness, pruritus, cerumen and purulence were graded. Swab specimens were taken for cytological examination Otitis externa (OE) in dogs is caused by a variety of and bacteria were counted and recorded on a scale of 0 aetiological agents. OE is commonly treated with (rare) to 3+ (numerous). Results showed that chronic

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 Abstracts 79 ear infections were being controlled at approximately P-63 two to three weeks into these clinical trials. Further- more, ZYMOX Otic solution was effective in improv- An atypical presentation of ototoxicity caused ing clinical signs associated with infectious OE. The by an ear medication containing florfenicol, grade of redness, pruritus, cerumen and purulence terbinafine and betamethasone acetate decreased within the first week, along with decrease of M. MASCARENHAS OE infection. Therefore, ZYMOX Otic solution was Promove Clınica Veterinaria, Rio de Janeiro, Brazil shown to be a viable treatment option for reducing bacterial and yeast infections and providing clinical Florfenicol, terbinafine and betamethasone acetate in improvement of OE in dogs. an adaptable gel (Osurnia, Elanco Animal Health; Source of funding: Self-funded. Greenfield, MA, USA) may be ototoxic and should Conflict of interest: None declared. only be applied with an intact tympanic membrane. Adverse effects associated withear medications may include otitis media, Horner’s syndrome, facial nerve paralysis and vestibular clinical signs (ataxia, nystag- P-62 mus, head tilt). The objective of this report was to describe a case of an atypical presentation of ototoxic- Video-otoscopy-guided tympanostomy tube ity caused by Osurnia not previously reported in the lit- placement for treatment of primary secretory erature. A 3-year-old male Maltese dog presented to an otitis media in cats emergency service with severe exudate, swelling, pain and oedema of the left eye. The signs appeared approx- R. LUCAS*, I. QUIRICO*, M. MASCARENHAS† imately 24 h after the dog received an application of and D. BEVIANI* Osurnia in both ears for treatment of acute otitis *Dermatoclinica, Sao~ Paulo, SP, Brazil; †Promove externa. The dog was diagnosed with neurogenic kera- Clınica Veterinaria, Rio de Janeiro, Brazil toconjunctivitis sicca (KCS) and an ipsilateral dry nose. Primary secretory otitis media is an uncommon disease There were no signs of other neurological deficits. in cats. Current treatment recommendations include Video-otoscopy was performed and the tympanic mem- manual removal of the mucoid effusion from the tym- brane of the left ear was ruptured. For treatment of panic cavity through a myringotomy incision and topi- this patient it was essential to clean and remove all of cal or systemic corticosteroids. Although some cats still the medication instilled within the external ear canal. present a recurrent mucoid effusion from the middle When neurogenic KCS is associated with an ipsilateral ear. The aim of this study was to describe video-oto- dry nose, it is likely that damage has occurred to the scopy-guided tympanostomy tube placement in four preganglionic parasympathetic fibres proximal to the cats with middle ear effusion and assess the clinical out- pterygopalatine ganglion. This is the first report of an come. Diagnosis of primary secretory otitis media was ototoxic medication that led to KCS and an ipsilateral based on history, physical examination, video-otoscopy dry nose. Considering that most cases of ear disease in and CT or MRI findings. Three cats presented bilateral dogs are treated topically, veterinarians should be and one unilateral otitis media. One showed concomi- aware of all possible signs of ototoxicity. tant respiratory signs and all cats presented with neuro- Source of funding: Self-funded. logical signs. The decision to add tympanostomy tube Conflict of interest: None declared. placement to the treatment was the recurrence of the otitis media 45 to 50 days after a prior myringotomy. A tympanostomy tube with 1.2 mm internal diameter (Shepard ventilation tube, Medicone; Cachoeirinha, P-64 Brazil) was placed in the myringotomy site under video-otoscopic guidance using malleable grasping for- In vitro activity of Copaiba oil against ceps (Karl Storz Veterinary Endoscope; Goleta, CA, Malassezia pachydermatis strains from canine USA). Clinical signs resolved rapidly in all cases fol- otitis lowing the procedure and all cases were asymptomatic A. PACHECO, S. SOUZA, L. MEDEIROS, E. at follow-ups at 30, 60, 90, 120 and 180 days. No SOUZA and T. SILVA adverse effects were noted. Results of this study indi- Center for Biological and Natural Sciences, Federal cate that tympanostomy tubes provide continual tym- University of Acre, Rio Branco, AC, Brazil panic cavity ventilation and drainage, and may be an effective alternative to repeated myringotomy for the Otological infections caused by Malassezia spp. are fre- treatment of primary secretory otitis media in cats. quent and the use of topical antimicrobials is used Source of funding: Self-funded. widely to control these infections. However, the devel- Conflict of interest: None declared. opment of resistance to antimicrobials has been reported widely and research for new therapy options is necessary. In this context, medicinal plants such as copaiba oil offer a promising alternative to control infections caused by several micro-organisms including Malassezia strains. Thus, we hypothesized that copaiba oil could inhibit yeast strains isolated from canine infections. For this study, a total of 23 strains of M. pachydermatis obtained from the culture of canine otitis were used. Initially, inhibition of growth was confirmed by the well diffusion technique using 100% copaiba oil. Subsequently, different concentrations of

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 80 Abstracts copaiba oil were assessed using the microdilution P-66 method to obtain the minimum inhibitory and fungici- dal concentrations (MIC and MFC). Fluconazole was Variation in biofilm production by used as a positive control. The MIC of copaiba oil in Pseudomonas aeruginosa isolates from canine the evaluated samples was 0.61%, confirmed by the otitis using different media and incubation absence of colonies through visual analysis. The MFC times was 1.85% because there was no yeast growth after inoculation. Thus, we can confirm that copaiba oil has M. L. PICCIONE and D. J. DEBOER the capacity to inhibit strains of M. pachydermatis School of Veterinary Medicine, University of Wisconsin- in vitro; however, clinical studies should be carried out Madison, Madison, WI, USA in order to confirm in vivo effectiveness. Pseudomonas aeruginosa is commonly isolated from Source of funding: Self-funded. dogs with otitis and frequently produces biofilms. There Conflict of interest: None declared. are vast differences in reported prevalence of biofilm- forming ability of P. aeruginosa isolated from canine otitis. Differences could be explained by strain, incubation time, temperature, media or regional variation. We P-65 sought to compare biofilm production of P. aeruginosa Reacidification of the ear canal with an ear isolated from canine otitis using different media (minimal versus enriched) at different incubation times. The cleanser in a model of alkalinization of the P. aeruginosa isolates were assessed for biofilm produc- external auditory canal in dogs tion using a crystal violet microtiter plate assay. Biofilm P. PANZUTI*, O. FANTINI†, M. MOSCA*, G. production in Lennox Broth (LB) and in M63 and M9 NOEL* and D. PIN* minimal media was assessed after 24 h, 48 h and 72 h † ° = *VetAgro Sup, Marcy l’Etoile, France; Vetoquinol, of incubation at 30 C. All isolates tested (n 29) were Lure, France classified as biofilm formers; however, the amount of biofilm produced varied greatly between strains. At Cerumen plays a protective role in the external auditory 24 h, all strains produced biofilm in LB. One strain did canal (EAC), especially through bactericidal and fungi- not produce biofilm in minimal media (M9, M63). cidal functions. In humans, changes towards alkalinity Incubation times >24 h in LB impeded accurate quan- of the EAC is one of the local factors in the progres- tification of biofilm production. Our study demon- sion of acute to chronic otitis. Higher EAC pH is strates that the amount of biofilm produced varies with found in dogs with otitis externa. We developed a time and medium used for incubation. We determined model of alkalization of the EAC in dogs in order to that optimal production of P. aeruginosa biofilm was evaluate the acidic effect of the ear cleaner Sonotix achieved with LB at 24 h. Although minimal media can (Vetoquinol; Lure, France). The study was approved be used, strains may be inaccurately categorized as non- under Ethic Committee no. 1812. Five mL of a pH biofilm formers. Further studies are needed to assess 10.1 phosphate-buffered saline (PBS) solution were factors increasing formation of biofilm and investigate instilled into both ears of 10 healthy beagles in order to biofilm-disrupting compounds for successful treatment reach pH 9. Then 2 mL of Sonotix was instilled into of canine Pseudomonas otitis. the right ear with the left ear kept untreated. Using a Source of funding: Self-funded. skin pH probe applied on the cavum conchae, EAC pH Conflict of interest: None declared. measurements were performed before the pH 10.1 PBS solution application, immediately after Sonotix application and at 1 h, 2 h, 3 h, 4 h, 9 h, 24 h and 48 h post- application. Overall, pH values were significantly lower for Sonotic-treated ears versus untreated ears (general- P-67 ized linear mixed model, P < 2 9 1016). Immediately Clinical efficacy of Neo-Burow’s solution in after Sonotix application, mean pH values dropped six dogs with suppurative otitis media abruptly to 6.38 + 0.27 in treated ears and then slowly † † decreases over the next 48 h until reaching pre-alkalini- A. ZAIKE*, K. MORI , Y. OSHIMA-TERADA and N. MURAYAMA† zation of 5.87 + 0.66. Sonotix elicited a quick reacidi- † fication of the EAC. This procedure appears suited for *Ishii Animal Hospital, Chiba, Japan; Dermatology evaluating the acidic effects of ear cleansers. Services for Dogs and Cats, Tokyo, Japan Source of funding: Vetoquinol. Suppurative otitis media is common in dogs, yet diffi- Conflict of interest: OF is an employee of Vetoquinol. cult to control owing to antimicrobial-resistant bacteria. Burow’s solution exhibits antibacterial activity for suppurative otitis externa and otitis media in human. How- ever, it takes several days to make and quality-control is challenging. Recently, Neo-Burow’s solution has been reported that has an aluminium acetate base; it is easier to prepare and has equivalent efficacy as Burow’s solution in humans. This case report assesses the efficacy for treating suppurative otitis media in dogs. The study included six dogs aged from three months to 14 years. They were diagnosed with suppurative otitis media based on typical clinical features with video otoscope and ear-discharge cytology. We irrigated ear canals and middle ears with Tritz-EDTA (Tris-EDTA Otic PLUS,

Kirikan Ltd.; Tokyo, Japan) and 0.05% chlorhexidine However, four ears of two dogs did not improve and gluconate solution (0.05% MASKIN WATER, Mar- the ceruminous cytological results showed bacteria. No uishi Pharmaceutical; Osaka, Japan) using a tube until adverse effects were observed during the trial. Our discharge disappears, and then rinsed them with Neo- results indicated Neo-Burow’s solution could be effec- Burow’s solution on the first day, and instilled five tive and safe in the treatment of chronic suppurative drops of five-fold diluted Neo-Burow’s solution into otitis media in dogs. the ear canals once daily for 14 days. Four ears of four Source of funding: Self-funded. dogs completely resolved and there were no bacteria Conflict of interest: None declared. with ceruminous cytological results after the treatments.

P-68 collected questionnaires, only 96 clinics (15.6%) had knowledge of Straelensia and 36 (6.7%) had diagnosed Larvicidal efficacy of nitenpyram against cases. Of 27 clinics reporting case numbers, 11 only infestation by the human bot fly in a dog: a saw one case while two had >20 cases. Of 26 clinics case report commenting, disease incidence was considered stable in P. ANDRIOTTI*, P. OLIVEIRA*, R. 69%, decreasing in 12% and increasing in 19%. The TORTORIELLO*, C. BOTELHO*, R. MELO† and J. first diagnosis of straelensiosis was made before 2011 in FERNANDES† nine clinics and after 2011 in 15. The distribution of *Post Graduation Program, Universidade Federal Rural cases covered 21/95 departments (autochthonous in ≥ do Rio de Janeiro, RJ, Brazil; †Department of 18). The mentioned techniques for diagnosis were deep Veterinary Science, Universidade Federal Rural de Rio “excisional” scrapings in 63% and histopathological in – de Janeiro, RJ, Brazil 76%. During 2011 2017, 46 canine cases were identified in LAPVSO (versus 49 during the 11 previous years). Injuries caused by human bot fly Dermatobia hominis In this study, 13 new locations (departments) were (Diptera: Cuterebridae) are extremely uncomfortable detected (seven from questionnaire, five from the labo- and painful for dogs. This species of fly causes nodular ratory and one from both). Straelensia remains poorly and furunculous primary cutaneous myiasis, which may known by veterinarians and thus is likely underdiag- result in opportunistic bacterial infections and predis- nosed. It continues to be present in its initially reported pose toward secondary myiasis. In spite of the wide area and is spreading peripherally with detection in five options of chemical treatment for large animals, canine distant areas: since 2017, two new areas have been dermatobiosis therapy remains a challenge for veteri- detected resulting in its presence in at least 39/95 narians. The aim of this case report was evaluation of departments of France. larvicidal efficacy of nitenpyram against infestation by Source of funding: Self-funded. human bot fly in a dog. A canine from Brazilian rural Conflict of interest: None declared. area was examined with a total of 16 oedematous, erythematous and ulcerated nodules. For confirmation of parasitism, all nodules were examined through digital- digital compression for visualization of D. hominis lar- P-70 vae. The treatment instituted was nitenpyram, at a dose of 1 mg/kg per os as a single dose. A 6 h post-adminis- Fleas in dogs: yearly evolution through tration assessment confirmed that all larvae were dead. prospective/retrospective evaluation over a 24- However, they were not expelled from the furuncular year period (1995–2018) lesions, likely due to their shape, large size relative to P. BOURDEAU*, P. BLUMSTEIN†, A. ROUSSEL‡, the opening of the tracts and the presence of rows of § spines on the larvae. Therefore, the dead larvae were F. TRAVERS , L. JEANNEAU* and V. BRUET* *E´cole Nationale Ve´te´rinaire de Nantes - ONIRIS, mechanically removed with minimal discomfort shown † by the dogs at the time of removal. It is concluded that University of Nantes, France; Veterinary Clinic Vetania, Vigneux de Bretagne, France; ‡Veterinary nitenpyram is effective in killing D. hominis and is use- § ful as an adjunct in treating dogs with myiasis caused Clinic Armonia, Villefontaine, France; Charles Lechat, by these larvae. Nantes, France Source of funding: This study was financed in part by The aim of this study was an analysis of the prevalence Coordenacßao~ de Aperfeicßoamento de Pessoal de Nıvel of flea infestation in dogs in western France. Data were Superior – Brazil. collected as recorded from the clinic of the Oniris DPM Conflict of interest: None declared. Unit. Every dog was assessed systematically for flea infestation (gross examination, combing methods) for the presence of fleas and/or flea faeces, possibly associated with hypersensitivity. Cases of dogs suggestive of P-69 having flea bite hypersensitivity (no flea detected = FBH) also were collected in a prospectively validated Current status of canine straelensiosis Good Clinical Practice system. We present herein (Straelensia cynotis) in France, its place of yearly variations over a retrospective 24-year period origin (1995–2018). Of 32,238 dogs, 2,804 were found infested P. BOURDEAU*, F. DEGORCE†, C. DOUINE* and (8.69%) and 3,858 (11.96%) were diagnosed with FBH F. CARREZ* resulting in a global prevalence of 20.6%. This was *E´cole Nationale Ve´te´rinaire de Nantes - ONIRIS, associated with a global flea:no flea ratio (f:nf) of 0.72. University of Nantes, France; †Laboratoire d’Anatomie The annual infestation ranged from 4.8% (2005) to Pathologique du Sud-Ouest LAPVSO, Toulouse, France 17.3% (1997) and, for FBH, from 17% (2000) to 34.8% (2012). The global impact of flea-related der- Since the first observations in the mid-1990s in France matoses decreased from 1997 to 2006 (minimum 8.2%), and presentation of the disease at WCVD 2000, strae- then increased (maximum 41.2% in 2012). A decrease lensiosis has been noted in several European countries. in flea detection was observed during the last period This study evaluated the current situation in France 2014–2018 (8.2 to 3.2%; possibly an isoxazolines using two mechanisms: (i) a questionnaire sent from effect?) and not for FBH frequency (23.8 to 22.3%). Laboratory DPM to veterinary practices and (ii) a ret- Until 2006, f:nf was >1 and remained 0.35 after 2013. rospective analysis of diagnostics performed in Although fleas have become less often detected in dogs, LAPVSO, both covering the period 2011–2017. Of 616

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 Abstracts 83 their impact remains high with approximately 30% of P-72 dogs having fleas and/or signs of FBH. These results question the quality of use in the field for modern Efficacy of sarolaner on the treatment of insecticides and the real clinical benefits obtained. myiasis caused by Cochliomyia hominivorax in Source of funding: Self-funded. dogs Conflict of interest: None declared. F. CARVALHO*,¶, M. SILVEIRA†, M. SOUZA§,M. MACHADO‡, C. ROCHA§ and J. FERNANDES‡ *College of Veterinary Medicine, Centro Universitario de Valencßa, Valencßa, RJ, Brazil; ¶College of Veterinary P-71 Medicine, Universidade Iguacßu, Nova Iguacßu, RJ, Brazil; † Fleas in cats: yearly evolution through Hospital Veterinario Sao~ Lazaro, Volta Redonda, RJ, Brazil; §Policlınica Veterinaria SOS Animal, Vassouras, prospective/retrospective evaluation over a 24- ‡ – RJ, Brazil; Department of Veterinary Science, year period (1995 2018) Universidade Federal Rural do Rio de Janeiro, P. BOURDEAU*, P. BLUMSTEIN†, A. ROUSSEL‡, Seropedica, RJ, Brazil § F. TRAVERS , L. JEANNEAU* and V. BRUET* The aim of this study was to evaluate efficacy of saro- *Department of Clinical Sciences, Unit DPM, laner on the treatment of myiasis caused by New World Veterinary School (Oniris), University of Nantes, † screwworm Cochliomyia hominivorax in naturally France; Veterinary Clinic Vetania, Vigneux de infested dogs. Selected for study were 12 dogs, of sev- Bretagne, France; ‡Veterinary Clinic Armonia, § eral breeds, aged 2- to 10-years old, with diagnosed Villefontaine, France; Charles Lechat, Nantes, France with myiasis by observation of maggots in wound(s). The objective of this study was to determine the preva- The dogs received the isoxazoline sarolaner (Simparica, lence of flea infestation in cats in western France. Data Zoetis; Parsippany, NJ, USA) as a single dose per os, were collected as recorded from the clinic of the Oniris following the manufacturer’s recommended dose of DPM Unit. Every cat was systematically assessed for 2 mg/kg (range: 2–4 mg/kg) for control of the cat flea flea infestation (gross examination, combing methods) and the brown dog tick (Rhipicephalus sanguineus)in for the presence of fleas and/or flea faeces possibly dogs. For 72 h post-treatment, the dogs were kept in associated with hypersensitivity. Cases of cats sugges- individual kennels to evaluate the number of dead lar- tive of having flea bite hypersensitivity (no flea detected vae. The regions most affected were: dorsum (43%), = FBH) also were collected in a prospectively validated nasal cavity (15%), ear (14%) and mammary tissue Good Clinical Practice system. We present here yearly with tumour (7%). A single dose of sarolaner had an variations over a retrospective 24-year period (1995– efficacy of 100% after 16 h post-treatment. No adverse 2018). Of 5,219 cats, 1,245 were infested (23.8%) and effects were observed in treated animals. Following 2,109 (33.5%) were diagnosed with FBH resulting in a treatment the animals returned to the environment in global prevalence of 64.6%. The range of annual infes- which they lived, and no new infestations were tation varied from 10.2% (2018) to 44% (2008) and, observed for a period of ≥30 days. Sarolaner is an for FBH, from 2.6% (2001) to 59.6% (2012). The glo- effective treatment option in myiasis caused by C. ho- bal impact of flea-related dermatoses progressively minivorax in dogs. decreased from 1995 to 2005 (22.7%), then increased to Source of funding: Self-funded. 72.7% in 2014 and then decreased to 59.7% in 2018 Conflict of interest: None declared. again (with a decrease of flea-positive cats). Interest- ingly, from 1995 to 2008 (apart from two years) a majority of cats were positive for fleas, yet during the latter 11 years fleas were not detected on most cats P-73 (maximum 83.2% in 2018). These results show that fleas persist as a dominant component of feline derma- Efficacy of imidacloprid plus moxidectin spot- tology (almost two thirds of cats examined). They also on and selamectin spot-on formulations against suggest that the flea–cat relationship may have been the ear mite Otodectes cynotis in dogs modified, resulting in a higher proportion of sensitized T. FUKASE* and Y. NAKAMURA† cats presented to dermatologists. *Department of Veterinary Medicine, Faculty of Source of funding: Self-funded. Veterinary Medicine, Okayama University of Science, Conflict of interest: None declared. Imabari, Japan; †Katsuragi Institute of Life Sciences, Tokyo, Japan The ear mite Otodectes cynotis inhabits the external ear canal of many species of mammals including dogs, cats and ferrets. We evaluated and compared the efficacy of two ectoparasiticides formulated as spot-on products against O. cynotis in dogs. Eighteen dogs naturally infested by O. cynotis were randomly assigned to three test groups of six animals each; an unmedicated control, one group medicated with imidacloprid + moxidectin spot-on and one group medicated with selamectin spot-on. The drugs were administered to the dogs six times at monthly intervals, with doses of 10 mg/kg for imidacloprid, 2.5 mg/kg for moxidectin, and 6 mg/kg for selamectin. The existence or absence

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 84 Abstracts of the mites in earwax was examined microscopically at provides the clinician with therapeutic options for med- weekly intervals. The dogs were all treated under the ical management. Guidelines for Proper Conduct of Animal Experiments Source of funding: Self-funded. and Related Activities in Katsuragi Institute of Life Conflict of interest: None declared. Sciences. Mites were detected from all of the dogs of unmedicated control throughout the six month research period and had completely disappeared from 6/6 dogs of the imidacloprid + moxidectin-medicated group by P-75 four months after the first treatment, and from 4/6 dogs of the selamectin-medicated group during the Search for cat fleas with low sensitivity research period. No adverse effects were observed in against fipronil in Japan any of dogs. The imidacloprid + moxidectin product Y. NAKAMURA* and T. FUKASE† was considered to be much more effective against *Katsuragi Institute of Life Sciences, Tokyo, Japan; O. cynotis in dogs than the selamectin products. †Department of Veterinary Medicine, Faculty of Source of funding: Self-funded. Veterinary Medicine, Okayama University of Science, Conflict of interest: None declared. Imabari, Japan The phenylpyrazole-class compound fipronil has been used widely as an insecticide. However, occurrence of fipronil-resistant cat fleas is suspected. The present P-74 study was done to find fipronil-resistant cat fleas in Leishmania mexicana in a central Texas cat: Japan. Cat fleas were collected from 10 dogs and 10 clinical presentation, molecular identification, cats every other year during 2010–2018. The fleas of sand fly vector collection and novel each isolate were examined for their sensitivity against fipronil through contact with the compound impreg- management nated on round filter paper, diameter 9 cm, in of 1, 5, K. HOPKE*, A. MEYERS†, L. AUCKLAND†,S. 10, 50, 100 or 500 lg amounts. Knock-down of the HAMER†, D. FLORIN‡, A. DIESEL* and A. fleas was observed over time for 24 h. Knock-down of PATTERSON* fleas from many isolates occurred dose-dependently *Department of Small Animal Clinical Sciences, College with 1–500 lg fipronil. However, a small number of the of Veterinary Medicine, Texas A&M University, College fleas of some isolates showed relatively low-sensitivity Station, TX, USA; †Department of Veterinary with no knock-down with 1–10 lg (one in 2010, two in Integrative Biosciences, College of Veterinary Medicine, 2012, one in 2014, two in 2016 and three in 2018). Texas A&M University, College Station, TX, USA; However, even the fleas of low-sensitivity isolates were ‡Independent Medical Entomologist, New York, NY, thought to be eliminated from body surfaces of dogs USA and cats after administration of a spot-on formulation Feline leishmaniasis is a rare parasitic disease that car- of fipronil, based on the results of another study which ries both therapeutic challenges and zoonotic concerns. examined the amounts of fipronil distributed on the This case report documents the clinical appearance, skin of the medicated animals. Accordingly, a fipronil diagnosis and novel treatment of a cat from central spot-on product should be considered still effective Texas with cutaneous leishmaniasis. The cat presented against cat fleas on dogs and cats in Japan. with a linear erosion on the right pinnal margin, an Source of funding: Self-funded. ulcerated exophytic nodule on the right hock and a Conflict of interest: None declared. swelling in the right nostril. Cytological and histopathological findings were consistent with leishmaniasis. PCR confirmed the presence of Leishmania mexicana, a species endemic to Texas. An epidemiological investigation P-76 was conducted by trapping sand flies from the cat’s Efficacy of some spot-on-formulated environment. Sand flies collected were identified as Lut- ectoparasiticides against the ear mite zomyia species, known vectors of Leishmania. Surgical excision of lesions is typically considered ideal, yet the Otodectes cynotis in cats lesion location in this case and the owner’s reluctance Y. NAKAMURA* and T. FUKASE† to pursue invasive treatments deemed surgical removal *Katsuragi Institute of Life Sciences, Tokyo, Japan; impractical. Given the lack of validated medical thera- †Department of Veterinary Medicine, Faculty of pies for L. mexicana in cats, treatments typically pre- Veterinary Medicine, Okayama University of Science, scribed for canine leishmaniasis were administered. Imabari, Japan Although allopurinol achieved clinical success, it was Otodectes cynotis inhabits the external ear canal of discontinued owing to suspected drug-related neutrope- many species including dogs, cats and ferrets. We eval- nia. Topical imiquimod also improved lesional skin yet uated and compared the efficacy of some ectoparasiti- was unsustainable owing to application difficulty. cides formulated as spot-on products against O. cynotis Alternative therapies were explored including an anti- in cats. Thirty cats naturally infested by O. cynotis were malarial Chinese herb known as artemisinin. Oral randomly assigned to five test groups of six animals administration of artemisinin resulted in nearly com- each; (i) an unmedicated control and four groups medi- plete clinical resolution of cutaneous lesions without cated with spot-on formulations of (ii) imidacloprid reported adverse events. The cat remained systemically (10 mg/kg) + moxidectin (1 mg/kg), (iii) selamectin healthy with stable lesions nearly eight months after ini- (6 mg/kg), (iv) selamectin (6 mg/kg) + sarolaner (1 mg/ tiation of artemisinin therapy. This case report demon- kg), and (v) fluralaner (40 mg/kg). Medications were strates feline leishmaniasis in central Texas and

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 Abstracts 85 administered to cats six times at monthly intervals, P-78 except for the fluralaner product which was administered twice at three month intervals. The existence or Pathological characterization of buffalo fly absence of mites in earwax was examined microscopi- lesions in Queensland cattle cally at weekly intervals. The cats all were treated M. N. NASEEM*, A. RAZA*, C. under the Guidelines for Proper Conduct of Animal CONSTANTINOIU‡, M. MCGOWAN†,R. Experiments and Related Activities in Katsuragi Insti- ALLAVENA†, A. TABOR* and P. JAMES* tute of Life Sciences. Mites existed in all of the unmedi- *Queensland Alliance for Agriculture & Food Innovation cated control cats through the test period and had (QAAFI), The University of Queensland, Australia; disappeared completely from all cats of the imidaclo- †School of Veterinary Science, The University of prid + moxidectin- and the fluralaner-medicated groups Queensland, Australia; ‡College of Public Health, by three and two months after the first medication, Medical & Vet Sciences, James Cook University, respectively. In both the selamectin- and the Queensland, Australia selamectin + sarolaner-medicated groups, mites were Haematobia irritans exigua, commonly known as buf- only eliminated from three cats during the research per- falo fly (BF), is the major ectoparasite of cattle in trop- iod. No adverse effects were observed in any of cats. ical and subtropical areas of Australia. BF also can The imidacloprid + moxidectin and fluralaner products transmit a filarial nematode, Stephanofilaria spp., which were considered to be much more effective against has been speculatively implicated in the aetiology of BF O. cynotis in cats than the selamectin and the lesions. Lesions associated with BF infestation range selamectin + sarolaner products. from raised, dry, alopecic, hyperkeratotic or scab- Source of funding: Self-funded. encrusted to variably haemorrhagic areas of ulceration Conflict of interest: None declared. which also represent a major animal welfare concern. Lesions appear different in northern and southern Queensland cattle. This could be a consequence of differences in cattle breeds, differences in the prevalence P-77 or epidemiology of the nematode, or differences in the Cat fleas with low sensitivity against ecology of their vector. Skin samples were collected isoxazoline compounds in Japan from cattle slaughtered at abattoirs in northern and – † southern Queensland. Lesions were scored (0 4) macro- Y. NAKAMURA* and T. FUKASE scopically for the presence of haemorrhage, ulceration, *Katsuragi Institute of Life Sciences, Tokyo, Japan; † exudation, alopecia and skin thickness. Skin strips rep- Department of Veterinary Medicine, Faculty of resenting the centre, edge and peripheral normal tissue, Veterinary Medicine, Okayama University of Science, were dissected from each lesion, preserved in 10% neu- Imabari, Japan tral buffered formalin and processed by paraffin sec- Some isoxazoline-class compounds such as afoxolaner tioning and staining with haematoxylin & eosin. have been used recently as active ingredients in ectopar- Microscopically, lesions were scored (0–4) for the epi- asiticides for dogs and cats. We found a cat flea popu- dermal and dermal damage, and inflammatory charac- lation showing low sensitivity against isoxazolines and teristics. This poster describes differences in the gross examined the drug sensitivity of this flea population. and microscopic pathology of BF lesions from northern Two dogs, kept in the same house and infested by cat and southern Queensland cattle, with and without fleas, were treated with an afoxolaner tablet at the rec- Stephanofilaria spp. present. The results will help to ommended dose and their flea burdens did not clarify the pathology and pathogenesis of BF lesions decrease. A sarolaner tablet, subsequently administered and provide a basis for investigating genetic differences to the dogs at the recommended dose, also was ineffec- between cattle in susceptibility to lesion development. tive. In order to clarify the reason why these isoxazoli- Source of funding: Meat and Livestock Australia. nes were ineffective, we carried out two experiments Conflicts of interest: None declared. under the Guidelines for Proper Conduct of Animal Experiments and Related Activities in Katsuragi Insti- tute of Life Sciences. In the first experiment, another two dogs that had previously been administered a P-79 fipronil spot-on product were artificially infested by the fleas collected from the above-mentioned case dogs. In Pemphigus foliaceus-like drug reaction to the second experiment, the two case dogs were cleaned canine leishmaniosis treatment: a case report by shampooing to remove all fleas and then artificially H. L. PEREIRA, C. PELETEIRO and A. M. infested by cat fleas collected from other dogs. In both LOURENCß O experiments, the artificially infested fleas were con- Centre for Interdisciplinary Research in Animal Health, firmed to be completely eliminated. These results mean Faculty of Veterinary Medicine, University of Lisbon, that the fleas on the case dogs were sensitive to fipronil Lisbon, Portugal and also that the blood of the case dogs contained This case report describes a pemphigus foliaceus (PF)- afoxolaner and/or sarolaner. Accordingly, it was con- like drug reaction to allopurinol and meglumine anti- cluded that the cat fleas on the two case dogs showed moniate therapy in a dog treated for canine leishmanio- low sensitivity against isoxazoline compounds. sis (CL). A 2-year-old, male neutered mongrel dog Source of funding: Self-funded. presented with scaling alopecia, mild pruritus, weight Conflict of interest: None declared. loss, pale mucosae membranes and lymphadenomegaly. Laboratory findings included severe nonregenerative, normocytic and normochromic anaemia and leucocyto- sis with neutrophilia, hyperglobulinaemia without

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 86 Abstracts hypoalbuminaemia, elevated urea and proteinuria. P-80 Leishmania ELISA test was positive for leishmaniosis (titre >1/320). Both allopurinol (10 mg/kg twice daily) The effectiveness of neem (Azadirachta indica) and meglumine antimoniate (100 mg/kg once daily) shampoo against dermatitis caused by brown were initiated. Five days later the dog started develop- dog tick (Rhipicephalus sanguineus) in dog ing dermatological signs compatible with a cutaneous N. I. SITEPU adverse drug reaction. The patient presented febrile Animal Teaching Hospital, Udayana University, with multiple pustules and vesicles in the abdominal Denpasar, Bali, Indonesia region, thighs and thorax, and also crusts and pustules in both ears and on head. Cytological smear revealed Ectoparasites are a common cause of skin problems in neutrophils with acantholytic cells. Skin biopsy was dogs; the brown dog tick (Rhipicephalus sanguineus)is performed and drugs were discontinued. The the most common cause in Bali, Indonesia. Treatment histopathological report described skin hyperplasia with with synthetic chemical drugs may have harmful spongiosis and subcorneal pustule formation containing adverse effects. The use of traditional medicine has acantholytic cells among hypersegmented nondegener- become an option in some parts of the world including ated neutrophils. A similar pustule was identified in a Indonesia. Neem leaves are used as a traditional medi- follicular sheath. The histological changes were compat- cine and allegedly have antiparasitic activity as they ible with PF possibly triggered by an adverse drug reac- contain sulfur, flavonoids and tannins. This study used tion. Treatment for PF was started with a low dose of dogs that presented to the Animal Teaching Hospital at prednisolone (1 mg/kg) due to leishmaniosis. Nucleo- Udayana University, Denpasar, Bali, Indonesia. Each tides and active hexose correlated compounds also were treatment was replicated two times a week, over a initiated for CL treatment. With minimal response after range of two weeks to two months. The results showed seven days and a poor prognosis, owners choose to that in the first week and second week, neem shampoo euthanize the dog. To the best of our knowledge, this is did not have a significant result. This is because the the first case report of a PF-like drug reaction to allop- pharmacokinetics of neem shampoo require a longer urinol and meglumine antimoniate therapy in a dog. time to achieve its maximum effect. A significant effect Source of funding: Self-funded. of neem shampoo started to be seen in the third week Conflict of interest: None declared. of treatment and in the fourth week, showed more significant results. Assessment criteria included observation of skin problems and number of ticks found by combing the dogs. Source of Funding: Self-funded. Conflict of Interest: None declared.

P-81 established reference genes for equine leucocytes: glyc- eraldehyde-3P-dehydrogenase, hypoxanthine ribosyl- IL-31: a mediator of pruritus in horses transferase and succinate dehydrogenase complex N. CRAIG, R. WILKES, N. S. MUNGUIA, M. subunit A. Differences in IL-31 transcription between DORR and R. MARSELLA healthy and allergic horses were tested using a one- Department of Small Animal Clinical Sciences, College tailed Wilcoxon–Mann–Whitney U-test. The Spearman of Veterinary Medicine, University of Florida, rank correlation test was used to evaluate the correla- Gainesville, FL, USA tion between pVAS and IL-31 transcription. No signifi- Interleukin (IL)-31 is a mediator of pruritus and may cant difference in IL-31 transcription was identified = have a role in equine allergic reactions although no between groups (P 0.48006) and no significant corre- study has demonstrated that the injection of IL-31 lation was found between pVAS and IL-31 transcrip- = directly triggers pruritus in horses. Recombinant equine tion in the cultured leucocytes (P 0.87851). In IL-31 was produced and its effect evaluated in four conclusion, although IL-31 plays a role in pruritus, normal horses. A dose-escalation study was done inject- transcription is no different in the absence of allergic ing intradermally either saline (0.2 mL) or increasing stimulation between allergic and normal horses, and doses of recombinant horse IL-31 (hIL-31) on separate thus currently cannot be used as a disease severity mar- days (0.2 mL containing 3 µgor5µg, or 0.4 mL con- ker. Further studies are needed to evaluate cytokine taining 10 µg hIL-31). Seconds spent itching at the site response upon allergen stimulation in horses. of injection were recorded by live observers for 1 h pre- Source of funding: Intramural grant competition, Univer- injection and 4 h post-injection. After saline injection, sity of Florida. one horse itched for 2 s while the other three did not. Conflict of interest: None declared. After the lowest hIL-31 (3 ug, n = 1) dose, 6 s of itching were observed at 180 min post-injection. After 5 ug (n = 1), 317 s of itching were recorded (peaks at 40 and 180 min). After 10 ug (n = 2), one horse itched at the P-83 injection site for 15 s and spent a longer time appearing Histological and molecular classification of irritated without rubbing the injection site, and the equine sarcoid: a pattern-based approach other horse itched for 79 s. Two peaks were detected (at 60 and 260 min for one horse, and 125 and 165 min G. TURA*, L. GALLINA*, F. SAVINI*, G. † ‡ for the other horse). In conclusion, IL-31 can trigger AVALLONE*, A. E. DURHAM , ,R.M.LA † pruritus in horses and its effect is not immediate. The RAGIONE and B. BACCI* observed delayed effect possibly is linked to the release *Department of Veterinary Medical Sciences, University † of other mediators. Further studies are needed to inves- of Bologna, Italy; School of Veterinary Science, ‡ tigate the cascade of events triggered by IL-31 in University of Surrey, UK; Liphook Equine Hospital, horses. Liphook, UK Source of funding: Intramural grant competition, Univer- Although the histological features of equine sarcoids sity of Florida. are well-characterized, there is no documented associa- Conflict of interest: None declared. tion between the clinical types and their histological features. The study goal was to characterize the mor- phological spectrum of equine sarcoids and propose a histological classification based on the growth pattern P-82 in 112 sarcoids diagnosed in 86 horses from the UK. Investigation on IL-31 levels and correlations Clinical subtypes were as follows: nodular (n = 35), verrucous (16), fibroblastic (12) and occult (five). Cases with pruritus in allergic horses were classified histologically as verrucous (epidermal N. CRAIG, R. WILKES, N. S. MUNGUIA, M. finger-like projections), nodular (dermal nodule sepa- DORR and R. MARSELLA rated from the overlying epidermis) and mixed (combi- Department of Small Animal Clinical Sciences, College nation of both). Immunohistochemical (IHC) of Veterinary Medicine, University of Florida, investigation with S100, a-SMA, p53 and PCR for Gainesville, FL, USA bovine papillomavirus (BPV) 1, 2 and 13 were per- Interleukin (IL)-31 is a mediator of pruritus in humans, formed in all cases. Chromogenic in situ hybridization mice and dogs. Injection of recombinant equine IL-31 (CISH) was performed in 44 cases. Tumours were clas- = has elicited pruritus yet it is unknown whether the sified histologically as verrucous (n 25), nodular (32) severity of pruritus in allergic horses is correlated to and mixed (55), and further categorized based on circulating levels of IL-31 and whether leucocytes of fibroblasts architecture into interstitial (75), and fascic- allergic horses intrinsically (in the absence of allergenic ular (37). The following histological features were iden- stimulation) release more IL-31 than normal horses. In tified: “picket fence” formation (58), hyperkeratosis this study, blood from 20 healthy and actively pruritic (60), epidermal hyperplasia (53) and rete pegs (27). (n = 20) allergic horses was collected and peripheral Myofibroblastic differentiation was confirmed by IHC a blood mononuclear cells were isolated. Global pruritus for -SMA in 44 cases and positivity was subepithelial Visual Analog Scale score (pVAS, 0–10, 10 most severe) (13), intratumoural (23) or peritumoral (8). All cases was obtained for allergic horses. IL-31 mRNA from were negative for S-100 and p53. The presence of BPV- lysated leucocytes was measured by quantitative PCR. DNA was confirmed by PCR in 107 cases. Forty-two IL-31 transcription was normalized to previously cases were positive for CISH in fibroblast nuclei and

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 88 Abstracts cytoplasm, revealing strong signal intensity and diffuse sarcoids. Future studies will aim at correlating this clas- pattern. The proposed histological classification is sification with tumour behaviour. intended to improve the diagnostic accuracy of equine Source of funding: Self-funded. Conflict of interest: None declared.

P-84 with O. natricis infestation. We reviewed two specimens from a private collection: an albino male and female, Efficacy of oral fluralaner for the treatment of both 2-year-old, weighing 1,139 and 1,694 g, respec- the Caparinia tripilis mite in African pygmy tively. Both animals presented poor quality scales, hedgehogs mobile black spots, depression, moulting difficulty and J. GUZMAN*, D. DACAK†, R. ORDONEZ~ ‡,M. loss of appetite. A direct collection of mites was per- LUCERO§ and R. FERREIRA¶ formed with a cotton swab and acetate tape. Morpho- *Animal House Dermatologıa y Spa, Cartagena, logical characteristics observed were identified Colombia; †Clınica Veterinaria Diego Dacak, Asuncion, as O. natricis. Both patients were medicated by orogas- Paraguay; ‡Clınica Hospital Animalopolis, Guayaquil, tric tube with afoxolaner 2 mg/kg. A 28.3 mg tablet Ecuador; §Clınica Veterinaria Tacuary, Asuncion, was macerated in a ceramic mortar with 20 mL of Paraguay; ¶Dermatovet Cursos, Porto Alegre, RS, organic coconut oil, and the correct liquid dose was Brazil administered. Observations were made on day (D)1, D3, D10 and D30 post-medication. On D3 ectopara- Fluralaner is an orally administered isoxazoline that sites were no longer found on the snakes, and dead provides protection against fleas, ticks and mites in ectoparasites were observed in the enclosures. No mites dogs and cats; however, studies in African pygmy were observed on D10, dead ectoparasites were found hedgehogs (Atelerix albiventris) are very limited. This in one of the enclosures on D20, and no mites were case series evaluates the efficacy and safety of fluralaner present on the snakes or in the environments by D30. in hedgehogs with Caparinia tripilis. Ten registered No adverse events were reported in either animal. A hedgehogs (four males, six females) were chosen, with single oral dose of 2 mg/kg afoxolaner was effective for ages between five months and 1-year-old, and weight the treatment of O. natricis in these Burmese pythons. – range 225 440 g. The main clinical signs were intense Source of funding: Self-funded. pruritus (100%), seborrhoea (100%), erythema (90%), Conflict of interest: None declared. scabs (60%) and loss of spikes (60%). Caparinia tripilis infestation was confirmed in all patients by taking skin samples with scotch tape and direct observation under a microscope. All hedgehogs were treated with a single- P-86 dose oral fluralaner at 25–56 mg/kg (Bravecto, MSD Animal Health; Beaucouze, France). The response to Efficacy of oral sarolaner for treatment of treatment was monitored by dermatological examina- sarcoptic mange in three naturally infested tion and sampling on day (D)7, D14, D21, D30, D60 rabbits: a case report and D90. All animals showed decreased pruritus and R. VENTURO*, J. BAJWA†, A. ARCILA‡ and C. improved lesions from D1 onward. On D7 and D14 ¶ dead mites were observed microscopically; at D21, ROMERO † – D30, D60 and D90 no mites were observed. No adverse *Private Practice, Lima, Peru; VetDERM Clinic effects of the medication were observed. The present Veterinary Dermatology & Ear Referral Medical Clinic, Vancouver, Canada; ‡Private Practice, Lima, Peru; study confirms the efficacy of fluralaner in hedgehogs ¶ infected with C. tripilis. Veterinary hospital DERMAVET, Mexico City, Source of funding: Self-funded. Mexico Conflict of interest: None declared. Sarcoptes scabiei infestation is uncommon in rabbits, yet this parasite has zoonotic potential and increasing numbers of cases have been reported recently. Macro- cyclic lactones have proven effective against S. scabiei P-85 infestation in rabbits. We report the effectiveness of oral sarolaner (Simparica Chewable Tablets, Zoetis; Use of afoxolaner on two Burmese pythons Parsippany, NJ, USA) treatment in three unrelated rab- (Python molurus bivittatus) with a snake mite bits, naturally infested with sarcoptic mange. The (Ophionyssus natricis) infestation affected rabbits were all intact male rabbits that C. ROMERO*, B. A. FUANTOS†, G. SHEINBERG§, belonged to different owners and were diagnosed with A. CORDERO‡ and R. HEREDIA* S. scabiei upon microscopic identification by superficial *Dermavet, Iztapalapa, Mexico; †Fauna Veterinary skin scraping. Presenting clinical signs included intense Clinic, Nuevo Leon, Mexico; §Centro Veterinario pruritus (3/3), ear pinna scabs (3/3), nasal hyperkerato- Mexico, Mexico City, Mexico; ‡Vetderm, sis (2/3), paronychia (3/3), periocular scaling (1/3), Guadalajarara, Mexico anorexia (1/3) and alopecia (3/3). Cohabiting animals included dogs (2/3) and a cat (1/3). Owners also had Ophionyssus natricis is considered an important parasite dermatological lesions (2/3). The rabbits were treated in reptiles that has zoonotic potential. The use of isoxa- with sarolaner from 4 to 4.7 mg/kg per os once, result- zolines in reptiles has not been reported. This study ing in the resolution of clinical signs within three weeks. objective was to evaluate the efficacy of afoxolaner All three rabbits relapsed on days 50, 53 and 56, (NexGard, Boehringer Ingelheim; Duluth, GA, USA) respectively, after the single sarolaner dose administra- in two Burmese pythons (Python molurus bivittatus) tion. The three rabbits then were treated with two doses

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 90 Abstracts of sarolaner p.o. administered 30 days apart, resulting effective for treatment of S. scabiei in rabbits, at dose in resolution of clinical signs within two weeks without range 4–4.7 mg/kg p.o. administered twice, 30 days recurrence for ≥11 months. The oral sarolaner formula- apart. tion labelled for dogs was considered palatable to all Source of funding: Self-funded. three rabbits. No patient exhibited adverse reactions to Conflict of interest: None declared. the medication. Based on this pilot report, sarolaner is

P-87 Complete blood count and biochemistry test results were normal. Skin scraping, tape test, hair pluck and Nonepitheliotropic cutaneous lymphoma of Wood’s lamp examination were done. Skin scraping immunophenotype T in a domestic feline: a and tape test revealed Demodex canis and cocci; case report Wood’s lamp examination was negative and KOH M. ANDRADE*, H. SOUZA†, D. ARAUJO‡,T. microscopy was positive for Trichophyton spp. Initially, COSTA‡, F. PAIVA‡, S. SILVA*, C. BOTELHO‡ and we stabilized the dog with carprofen and Cosequin HA J. FERNANDES† (Bioiberica; Barcelona, Spain); primary treatment *Autonomous Veterinary, RJ, Brazil; †Post Graduation included antibiotics, and ketoconazole and ivermectin Program, Universidade Federal Rural do Rio de Janeiro, per os. However, there was minimal improvement. RJ, Brazil; ‡Department of Veterinary Science, Hence, abdominal ultrasound was suggested to search Universidade Federal Rural de Rio de Janeiro, RJ, for any underlying cause, although a recent ultrasound Brazil and magnetic resonance imaging performed by the previous veterinarian did not identify any abnormalities. Lymphoma is the most common malignant neoplasm The new abdominal ultrasound revealed two pelvic in cats; however, cutaneous involvement occurs in only masses. Exploratory laparotomy was performed. A tes- – 0.2 1.7% of affected animals. Cutaneous lymphoma ticular tumour was excised and the second mass was may occur in the epitheliotropic or nonepitheliotropic found to be prostatic tumour for which finasteride p.o. forms. Epitheliotropic lymphoma is mostly comprises T was recommended. Biopsy of the testes confirmed a lymphocytes and the nonepitheliotropic form usually nonseminomatous malignant germ cell tumour. Post- comprises B lymphocytes. This report describes a case surgery, the dog showed remarkable improvement in of nonepitheliotropic T-immunophenotype lymphoma skin and hair coat as well as overall health. This case in a cat in Brazil. The animal presented with multiple, illustrates a possible relationship between dermatitis, nodular, ulcerated lesions on the right flank and inner demodicosis and a testicular tumour in older dogs. right thigh, reporting a one-month evolution, associated Source of funding: Self-funded. with systemic clinical signs of hyporexia, weight loss Conflict of interest: None declared. and prostration for three months. Histopathological findings from biopsies from the affected areas were compatible with cutaneous lymphoma. The treatment was based on chemotherapy protocols, starting with P-89 Lomustine as sole agent, later changing to CHOP protocol, using vincristine sulfate, doxorubicin, cyclophos- Intratumoural treatment of a high-grade phamide and prednisolone. A partial response was perianal mast cell tumour with the novel small achieved, with improvement of the condition up to the molecule tigilanol tiglate (EBC-46). beginning of the second protocol cycle. After a period of absence of the tumour, a severe worsening of the G. K. BROWN, J. E. CAMPBELL, P. D. JONES, T. general state of the animal and the appearance of a R. DERIDDER and P. W. REDDELL new lesion was observed and the animal was eutha- QBiotics Group Limited, Yungaburra, Queensland, nized. Subsequently, immunohistochemical evaluation Australia confirmed nonepitheliotropic cutaneous lymphoma and A 12-year-old Staffordshire bull terrier was cytologi- identified immunophenotype T cells. With a small num- cally diagnosed with a high-grade perianal mast cell ber of published reports, the treatment represents a tumour. This tumour is associated with a poor progno- challenge and has no clear recommendations. The pro- sis and surgical excision was likely to result in an tocol generally described with the longest remission incomplete excision with a high risk of local recurrence. time is the Madison–Wisconsin protocol, which is The main objective was to treat this patient using intra- adapted to the CHOP protocol used for this cat. tumoural tigilanol tiglate (1 mg/mL in buffered 40% Source of funding: This study was financed in part by propylene glycol). Two injections were administered 32 Coordenacßao~ de Aperfeicßoamento de Pessoal de Nıvel days apart to locally treat the tumour. On treatment Superior-Brazil. day, the modified ellipsoid method was used to calcu- Conflict of interest: None declared. late tumour volume which was then used to determine tigilanol tiglate dose (50% v/v; 0.5 mg/cm3). Efficacy was assessed on the basis of presence/absence of a complete response (full tumour resolution) at day (D)28 P-88 and (D)60 using the Response Evaluation Criteria in Solid Tumours (RECIST) developed by the Veterinary A case report on nonseminomatous malignant Cooperative Oncology Group (VCOG). Before treat- germ cell testicular tumour induced dermatitis ment, the tumour was 2.7 cm in diameter and had a 3 M. AWALE tumour volume of 5.1cm . A wound with a maximum 2 Pet Wellness Clinic, Pimple Saudagar, Pune, surface area of 3.6 cm had formed by D7. At D28 a Maharashtra, India partial response was recorded and a tumour volume of 0.6 cm3, a volume reduction of 87%. On D32, a second A 9-year-old male Labrador retriever with a history of treatment was administered and a wound had formed previous cryptorchidism and castration, was presented with a maximum surface area of 6.3 cm2 four days with inability to rise, joint pain, erythematous and mul- later. By D60 after first treatment, wound healing was tifocal alopecia, crusts, scaling, mild pruritus, hyperpig- complete and a complete response was recorded. This mentation, pendulous prepuce and gynaecomastia.

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 92 Abstracts case report highlights the treatment of an intractable P-91 tumour on a patient with an otherwise poor prognosis using intratumourally administered tigilanol tiglate. Leucotrichia of epitheliotropic lymphoma in Name and reference number of ethics overview commit- five dogs tee: AEC Queensland, CA 2018-08-1223. † T. NARITA*, K. SHIBATA*, K. IYORI and N. Source of funding: QBiotics Group Limited. MURAYAMA‡ Conflict of interest: GKB, JEC, PDJ, TRD and PWR *DVMs Animal Medical Center Yokohama, Kanagawa, are employed by QBiotics Group Limited. Japan; †Vet Derm Tokyo, Kanagawa, Japan; ‡Dermatology Services for Dogs and Cats, Tokyo, Japan Canine epitheliotropic lymphoma has a poor prognosis P-90 and consists of several types of dermatological lesions including erythema, plaques, scaling, nodulation, ulcer- A preliminary study of intratumoural ation, alopecia and depigmentation of skin. Leu- administration of novel small molecule tigilanol cotrichia, not currently a clinical feature of tiglate (EBC-46) for treatment of canine soft epitheliotropic lymphoma, is caused by a variety of tissue sarcomas metabolic and inflammatory diseases and has been reported in association with vitiligo, alopecia areata, G. K. BROWN, J. E. CAMPBELL, P. D. JONES, T. lymphocytic folliculitis, post-inflammatory folliculitis R. DERIDDER and P. W. REDDELL and drug reactions. In this study, we observed leu- QBiotics Group Limited, Yungaburra, Queensland, cotrichia in five dogs with epitheliotropic lymphoma. Australia The dogs were patients of three referral hospitals and The primary treatment option for soft tissue sarcomas seen between 2016 and 2017. The breeds were two is surgical resection, yet this can be hampered by the Welsh corgis, two mixed breeds and a miniature dachs- size and location of the tumour. The main objective of hund. The ages at onset were 6–14 years (mean 8 this study was to provide an assessment of the safety years). The dogs had several typical skin lesions of and efficacy of intratumoural tigilanol tiglate (1 mg/mL epitheliotropic lymphoma including erythema, scaling, in buffered 40% propylene glycol) for the treatment of alopecia and nodules. In addition, all dogs had macular canine soft tissue sarcomas. This study was conducted leucotrichia, which appeared especially on the muzzle, under permit from the Australian pesticides and veteri- extremities and trunk. Histopathological findings nary medicines authority. On the day of treatment, revealed neoplastic lymphocytes infiltrating the epider- tumour volume was determined using the modified mis and adnexal structures, follicular walls (five) and a ellipsoidal method and tigilanol tiglate dosing was cal- nodular infiltration of the dermis (one). There were no culated based on this tumour volume (40% v/v; observations of infiltration into anagen hair bulbs as 0.4 mg/cm3). The required volume was drawn up into a seen in alopecia areata. Immunohistochemical evalua- Luer lock syringe and injected in a fractionated manner tion of neoplastic cells revealed CD3-positive T cells in as evenly as possible throughout the tumour. Dogs two dogs. Although the mechanism underlying the were excluded if the required tigilanol tiglate dose development of leucotrichia remains unknown, it was exceeded 0.25 mg/kg bodyweight or was >5 mg. expected that the regrowth of hair could turn white at Twenty-two dogs were treated and efficacy was assessed the site of early lesions in which epitheliotropic lym- on the basis of presence/absence of a complete response phoma cells infiltrated into the hair follicle. Therefore, (full tumour resolution) at day (D)28 and D42 using it is suggested that leucotrichia could be one of the clin- the Response Evaluation Criteria in Solid Tumours ical features of epitheliotropic lymphoma. (RECIST) developed by the Veterinary Cooperative Source of funding: Self-funded. Oncology Group (VCOG). A second tigilanol tiglate Conflict of interest: None declared. treatment was administered if a complete response was not achieved. Sixty-eight percent (15/22) of dogs achieved a complete response by D42. A second injection improved overall efficacy to 82% (18/22 dogs). P-92 This preliminary study demonstrated that tigilanol Linear arrector pili muscle hamartoma on the tiglate was well tolerated with good efficacy for local treatment of soft tissue sarcomas. tail of a Sphynx cat Source of funding: QBiotics Group Ltd. M. ORLANDI*, N. FANTON† and F. ABRAMO‡ Conflict of interest: GKB, JEC, PDJ, TRD and PWR *Laboratorio MyLav-La Vallonea, Passirana di Rho are employed by QBiotics Group Ltd. (MI), Italy; †Clinica Veterinaria Tibaldi, Milano, Italy; ‡Department of Veterinary Sciences, University of Pisa, Pisa, Italy Arrector pili muscles are specialized adnexal structures made of smooth muscle, rarely involved by disease; piloleyomioma and piloleyomiosarcoma are the unique examples of tumours of these muscles in cats. Arrector pili muscle hamartoma is described in humans and rarely in dogs. The aim of this presentation is to describe clinical and microscopic features of an arrector pili muscle hamartoma in a cat. A 2-year-old male Sph- ynx cat presented with nonpruritic, nonpainful tail nodules, noticed one month previously. Clinically, seven

5 mm, alopecic, dermal nodules were arranged linearly each time. Because of the good results, the authors sug- on the dorsal surface of the tail, surrounded by hyper- gest that further studies with toceranib phosphate as an trichotic skin. FNA cytological evaluation showed an adjuvant treatment in squamous carcinomas should be inflammatory process. No therapy was instituted. After done. two months, two new small nodules were observed. Source of funding: Self-funded. Surgical excision of three nodules was performed under Conflict of interest: None declared. general anaesthesia and submitted for histopathological evaluation. Histologically, unencapsulated proliferation of mature smooth muscle cells, organized in well- defined, randomly oriented and abnormally large bun- P-94 dles was seen, expanding pre-existent appendages and surrounded by lymphocytes. There was no evidence of Multicentric squamous cell carcinoma in situ cytological atypia or mitoses within the muscle compo- in a dog resembling Bowen’s disease nents. The diagnosis was arrector pili muscle hamar- B. L. SANZ RESSEL*,†, M. J. DUARTE‡,G.C. toma. The pseudo-Darier sign, characteristic of muscle BROGLIA¶,§, C. G. BARBEITO*,† and A. R. hamartoma, was negative. Two weeks later, biopsy sites MASSONE** had healed uneventfully and the other nodules had *Laboratorio de Histologıa y Embriologıa Descriptiva, undergone progressive reduction, yet hypertrichosis per- Experimental y Comparada (LHYEDEC), Facultad de sisted. We hypothesize an origin from arrector pili mus- Ciencias Veterinarias, Universidad Nacional de La Plata, cle because of the anatomical relationship with hair La Plata, Buenos Aires, Argentina; †Facultad de follicles. Linear distribution of arrector pili hamartomas Ciencias Veterinarias, Consejo Nacional de and associated hypertrichosis have been described in Investigaciones Cientıficas y Tecnicas (CONICET), humans. To the best of our knowledge, this is the first Universidad Nacional de La Plata, La Plata, Buenos case reported in a cat. Aires, Argentina; ‡Hospital Escuela de Pequenos~ Source of funding: Self-funded. Animales, Area de Clınica Medica en Pequenos~ Conflict of interest: None declared. Animales, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina; ¶Hospital Escuela de Pequenos~ Animales, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, La P-93 Plata, Buenos Aires, Argentina; §Catedra de Use of toceranib phosphate as an adjuvant Enfermedades de Caninos y Felinos Domesticos, Facultad de Ciencias Veterinarias, Universidad Nacional treatment for squamous cell carcinoma: a case de La Plata, La Plata, Buenos Aires, Argentina; report **Laboratorio de Patologıa Especial Veterinaria Dr. R. RAMADINHA, J. ALMEIDA and D. CAICO Bernardo Epstein, Facultad de Ciencias Veterinarias, ARAUJO Universidad Nacional de La Plata, La Plata, Buenos Clinical and Surgery Department - Rural Federal Aires, Argentina University of Rio de Janeiro, Seropedica, RJ, Brazil Multicentric squamous cell carcinoma in situ (MSCCIS) The authors describe the use of toceranib phosphate is very uncommon in dogs, with only one case reported (Palladia, Zoetis Animal Health; Parsippany, NJ, USA) in the veterinary literature. We describe an 8-year-old as an adjuvant therapy for squamous cell carcinoma of female mixed-breed dog that developed multifocal well- a 12-year-old female pitt bull dog. The tumour started circumscribed, pigmented, crusted, proliferative or ero- as multiple small and scaly nodules on the abdomen sive cutaneous lesions, measuring 0.5–3 cm in diameter. and pelvic limbs. The dog had several surgical excisions Lesions were found in pigmented and nonpigmented of nodules and was treated with topical imiquimod. A skin. The most frequently affected areas were dorsal larger tumour in the inguinal region grew and had an thorax, abdomen and proximal limbs. Metastasis was extensive surgical procedure. Subsequently, treatment not observed. The dog was otherwise healthy. with carboplatin (230 mg/m2) was done with no Histopathologically, lesions were characterized by well- response followed by radiation therapy (three fractions circumscribed areas of neoplastic keratinocytes without of 8 Gy each) that controlled the neoplasia for a short invasion through the basement membrane into the der- time. In June 2019, the authors decided to initiate an mis. Irregular epidermal and superficial follicular experimental and palliative therapy with intralesional hyperplasia and dysplasia were observed. Moderate toceranib phosphate (2.75 mg/kg) every 48 h. The parakeratotic hyperkeratosis also was detected. Lesions growth of the neoplasia slowed and the inguinal on pigmented areas showed marked hyperpigmentation tumour reduced in volume by approximately 50%. The of the epidermis. Papillomavirus-associated cytopathic dog was still in treatment, at the time of writing with effects in the keratinocytes were not observed. The case reduced doses (2.50 mg/kg) and palliative control of reported here is the second reported case of MSCCIS the neoplasia. Normochromic normocytic anaemia was in a dog with clinical and histological features of observed, and also episodes with diarrhoea occurred Bowen’s disease. four and eight months after starting therapy. The dog Source of funding: Self-funded. recovered after treatment was suspended for two weeks Conflict of interest: None declared.

P-95 (Nolvasan Surgical Scrub, Fort Dodge Animal Health; Fort Dodge, IA, USA) every other day. Another dog Mini review and update of lethal was treated with the same topical 2% chlorhexidine acrodermatitis of bull terriers acetate twice weekly. The dog without pyoderma was N. A. MCEWAN given a moisturizing shampoo twice weekly (Afloat Vet University of Liverpool, School of Veterinary Science, Moisturize Foam, QIX Corporation; Tokyo, Japan). Liverpool, Cheshire, UK All dogs’ pruritus and clamminess were resolved by these treatments, with no anti-inflammatory agents nec- Lethal acrodermatitis (LAD) is an autosomal recessive essary. Hyperhidrotic skin has been reported to have a genetic condition in bull terriers. The objectives of this high pH and the clamminess seen in this study was paper were to give an overview of LAD and to inform resolved without using anti-seborrhoeic shampoo. In that a DNA test is available. The first cases of LAD in conclusion, the clamminess of the skin of Yorkshire ter- bull terriers were reported in 1986. The largest study of riers may occur as a primary disease and may be more 28 cases was reported by McEwan, who discovered low attributed to hyperhidrosis than seborrhoea. immunoglobulin (Ig)A levels in these cases and pro- Source of funding: Self-funded. posed that LAD was hereditary and immunosuppres- Conflict of interest: None declared. sive. LAD causes growth stunting with most cases only achieving half their body size. The skin lesions consist of crusting and hyperkeratosis mainly over pressure points such as the elbows and hocks, the paws are splayed with hyperkeratosis of the foot pads and older P-97 cases have deformed claws. Bacterial and yeast skin Partial and full-thickness burns in three dogs infections (including Candida spp.) and upper respira- from an electric heating pad tory disease with nasal discharge are common with † LAD. The clinical signs and histopathological confir- V. BALAZS*, A. LOPEZ and B. RIVERA* mation of diffuse parakeratosis suggest zinc deficiency, *Oftaderm, Clınica especializada en Oftalmologıa y † and the disease was compared to lethal trait A46 of cat- Dermatologıa Veterinaria, Santiago, Chile; Centro tle and acrodermatitis enteropathica of humans. Both Vetera-Terapis, Temuco, IX Region, Chile of these conditions are responsive to zinc, yet attempts Prolonged exposure to electric heating pads in veteri- to treat LAD with zinc have all failed, including by nary clinics frequently causes thermal injuries in small peritoneal injections. Recently a DNA test for the gene animals. Three cases are described. A 1-year-old male splicing defect of MKLN1 has become available for German shepherd doe (Case 1) was anaesthetized for LAD (Animal Health Trust; Lanwades Park, Kentford, gastrointestinal surgery with a heating pad under the Newmarket, UK). In conclusion, LAD in bull terriers back. On Day 7 post-surgery, the dog showed a large is caused by a MKLN1 gene splicing defect and a DNA area of devitalized skin on the dorsal trunk that pro- test is now available. gressed to well-demarcated serpiginous ulcerations with Source of funding: University of Liverpool, Liverpool, central necrosis. These partial thickness lesions compro- UK. mised 10–20% of the body surface. Treatment was suc- Conflict of interest: None declared. cessful with Clindamycin 11 mg/kg per os every 12 h and a topical product made of a combination of honey and essential oils (Centro-Vetera; Temuco,Chile). A 1- year-old female mixed-breed dog (Case 2) presented P-96 eight days after ovariohysterectomy with demarcated necrotic lesions on the right flank surrounded by ery- Clamminess in Yorkshire terriers caused by thematous skin. The dog had been kept on lateral perspiration recumbency on a heating pad after surgery. Necrotic Y. OSHIMA-TERADA, K. MORI and N. skin was resected and subcutaneous tissue approxi- MURAYAMA mated with sutures. Partial-thickness lesions compro- Dermatology Services for Dogs and Cats, Tokyo, Japan mised 10% of the total body surface. Healing was achieved using DuoDERM Hydroactive Gel (Uru- Few studies have been reported on primary hyperhidro- farma; Montevideo, Uruguay). A 6-month-old female sis in dogs. The aim of this study was to report clammi- Pomeranian (Case 3) presented 14 days after an ovario- ness on the trunk which may occur as a primary hysterectomy with necrotic lesions on both flanks, dor- disease in Yorkshire terriers. Three pure-bred Yorkshire sum, and under the neck. The dog had been kept on a terriers and one Yorkshire terrier mix were presented heating pad for a long time after surgery. Full-thickness with clamminess on the trunk with pruritus. Three dogs injuries affected 20–30% of the body surface. The dog also showed papules and erythema on trunk caused by died due to systemic decompensation. The use of elec- superficial pyoderma, and 2/3 had recurrent pyoderma. tric thermal pads in veterinary clinics can lead to severe Pruritus and clamminess were persistent whether or not injury and should be avoided. the pyoderma existed. The age when the clinical signs Source of funding: Self-funded. first became apparent ranged from six months to 2 Conflict of interest: None declared. years. The mean pH of the skin on the dorsal thorax of all four dogs ranged from 9.0 to 9.7. Histopathological results showed sebaceous gland hyperplasia and dilated apocrine glands. Two dogs were treated with oral antibiotics and topical 2% chlorhexidine acetate

P-98 presented with a keratinized horn measuring 3.0 x 4.0 cm protruding from the snout, with multiple Sebaceous adenitis: effectiveness of ciclosporin warty papillomatosis lesions of the face and oral cavity. treatment, regardless of disease stage The animal was concomitantly diagnosed with Ehrlichia canis and treated with doxycycline hydrochloride, A. BLANCO*, M. LOIZA*, A. WOLBERG*, E. 5 mg/kg per os every 12 h for 30 days. After resolution TONELLI*, M. DUARTE*, M. SCARPA* and A. † of this haemoparasitosis, under general anaesthesia the DUCHENE *Dermatology, Small Animals Hospital, School of apical portion of the horn was resected and the base of the surgical wound was frozen, in two freezing cycles Veterinary, University of Buenos Aires, Argentina; †Pathobiology Service, Small Animals Hospital, School approximately 1 min each, with direct spray application. Cryosurgery was used on the other verrucous of Veterinary, University of Buenos Aires, Argentina lesions, with two cycles of freezing. After a 14 day There are different therapeutic options for the long- recovery, the cryosurgery was repeated on the remain- term management of sebaceous adenitis (SA) in dogs ing warty lesions and the remaining portion of the cuta- with variable results. This purpose of this study was to neous horn, in two freezing cycles similar to the first demonstrate the effectiveness of ciclosporin regardless application. After 10 days from the second session, all of the stage of the disease. This is a retrospective lesions were in complete remission. Six months after review of dogs with biopsy-confirmed sebaceous adeni- the procedure, the animal remained in complete remis- tis treated with ciclosporin from January 2009 to sion. The treatment typically indicated for cutaneous December 2019. Eighteen dogs were included. The fol- horns is surgical excision; however, in this case, cryo- lowing breeds were represented: akita inu (7), standard surgery was chosen because in the affected region surgi- poodle (3), chow chow (1), Lhasa apso (1), shih tzu (1), cal access was difficult. In conclusion, cryotherapy may German shepherd dog (1) and four mixed-breeds (13 be an effective tool for the management of cutaneous males, five females). Ages ranged from 1 to 8 years. horn secondary to papillomatosis if surgical removal is The predominant skin lesions were scales, follicular less desirable. casting and hypotrichosis. Three skin biopsies were Source of funding: This study was financed in part by taken from each patient. Histopathological results Coordenacßao~ de Aperfeicßoamento de Pessoal de Nıvel showed various stages of the disease, including absence Superior-Brasil (CAPES). of the sebaceous glands in 10/18 cases (55.55%), Conflict of interest: None declared. inflammation surrounding the glands in 5/18 (27.77%) and both characteristics in 3/18 (16.66%). Treatment with ciclosporin was initiated at 5 mg/kg per os once daily for 90 days, accompanied by baths every four P-100 days with a sulfur and 2% salicylic acid shampoo (Der- mosedan champu antiseborreico, Inst. Dermatologico; Evaluation of a plant-based balm in preventing Buenos Aires, Argentina), and daily sprays with 50% pedal lesions on sled dogs propylene glycol. Thirteen cases (72%) showed disap- † ‡ A. BOUVIER*, C. THORIN , H. POULIQUEN and pearance of scales and follicular casting with hair P. BOURDEAU* regrowth, four cases (22%) showed reduction in scale *Department of Clinical sciences, Veterinary School and follicular casting and partial hair regrowth, and (Oniris), University of Nantes, Nantes, France; one case (5%) had a partial response. It is concluded † Department of Management and statistics, Veterinary that ciclosporin and topical therapy are highly effective School (Oniris), University of Nantes, Nantes, France; in the treatment of sebaceous adenitis, regardless of the ‡ Department of Biology, Pathology and Nutrition disease stage. Sciences, Veterinary School (Oniris), University of Source of funding: Self-funded. Nantes, Nantes, France Conflict of interest: None declared. Sled dogs suffer from painful pododermatitis during the training season. The objective of this study was to evaluate the efficacy of Dermoscent BIO BALM (Labo- P-99 ratoire Dermo Cosmetique Animale; Castres, France) on foot pads of sled dogs. Fifty-two dogs were included Cryosurgery in the treatment of cutaneous in two groups: in A (22 dogs) all feet were treated; and horn associated with papillomatosis: a case in B (30 dogs), the treatment was applied on right foot report pads only (left legs as control), for a total of 148 foot pads treated and 60 control. The balm was applied F. PAIVA*, M. SOUZA*, L. JUNIOR†, T. COSTA*, ‡ before and after daily running (15–50 km) during 15 D. ARAUJO*, C. BOTELHO* and J. FERNANDES consecutive days. Evaluation of erythema, abrasions or *Post Graduation Program, Universidade Federal Rural cracks on all foot pads was made twice daily on days do Rio de Janeiro, RJ, Brazil; †Autonomous Veterinary, ‡ (D)1–2 and D15 in Group A; on D1–2, 4–5, 8–9, 12–13 RN, Brazil; Department of Veterinary Science, and 15 in Group B. Unpaired Student’s t-test was used Universidade Federal Rural de Rio de Janeiro, RJ, for statistical analysis. During the study, erythema/ Brazil abrasions and cracks were detected in respectively 285/ Cutaneous horns are keratinized acellular formations, 2,000 and 43/2,000 feet examined, representing in manifesting in association with other skin diseases. The Group B 7.3% of treated (versus 14.1% untreated) for occurrence is rare, with a frequency of 0.1% of skin erythema/abrasion and 0.5% of treated (versus 2.3% biopsies in dogs. This report describes a case of cuta- untreated) for cracks. The overall risk of erythema/ neous horn associated with papillomatosis treated with abrasion was significantly lower with treatment [odds cryosurgery. A 5-year-old male mixed-breed dog was ratio (OR) = 0.2984, P = 1.35e-7] and this difference

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 96 Abstracts increased with time, from D5 (OR = 0.0638, speeding the healing of feet during intensive training of P = 0.00741) up to D15 (OR = 0.0231, P = 9.50e-7). sled dogs. The treatment also reduced cracks (OR = 0.1272, Source of funding: Self-funded. Laboratoire Dermo Cos- P = 0.00143). The balm was well-tolerated by dogs and metique Animale provided free product upon request by also reduced “snow balls”, a frequent cause for sled authors and played no role in the study otherwise. dogs’ foot irritations. Dermoscent BIO BALM appears Conflict of interest: None declared. effective in limiting foot lesions on sled dogs during intensive training. Source of funding: Self-funded. Laboratoire Dermo Cos- metique Animale provided free product upon request by P-102 authors and played no role in the study otherwise. Conflict of interest: None declared. The use of interactive technology to enhance learning in clinical veterinary dermatology A. PUTRA*, W. GRAM*, C. STEFANOU*,† and D. SANTORO* P-101 *Department of Small Animal Clinical Science, Healing effect of a plant-based balm on pedal University of Florida, College of Veterinary Medicine, FL, USA; †Department of Family, Youth, and lesions on sled dogs Community Sciences, University of Florida, College of A. BOUVIER*, C. THORIN†, H. POULIQUEN‡ and Agricultural and Life Sciences, FL, USA P. BOURDEAU* Clinical teaching in veterinary medicine, including der- *Department of Clinical Sciences, Veterinary School matology, is very challenging. In several disciplines, (Oniris), University of Nantes, Nantes, France; † there is an increasing interest in the use of novel inter- Department of Management and Statistics, Veterinary active technology-based techniques, such as adaptive School (Oniris), University of Nantes, Nantes, France; ‡ learning, to provide students with more opportunities Department of Biology, Pathology and Nutrition to learn. Few studies have evaluated the use of this Sciences, Veterinary School (Oniris), University of technique in veterinary medicine. We hypothesized that Nantes, Nantes, France adaptive learning modules given during dermatology Pododermatitis is a frequent cause of pain and lame- rotation would improve students’ dermatology test ness of sled dogs due to training conditions. Regardless scores, and students would regard these modules as of protective boots that dogs wear to prevent lesion beneficial to their overall learning experience. Ten inter- deterioration, additional treatment is often necessary. active dermatology modules were developed using an The objective of this study was to evaluate the healing adaptive learning platform. Using a quasi-experimental effect of Dermoscent BIO BALM (Laboratoire Dermo design, incoming veterinary students to the dermatol- Cosmetique Animale; Castres, France) on lesions ogy rotation who agreed to participate were randomly induced during training. Thirty included dogs received assigned to treatment (given the modules during the an application of balm on right foot pads (left ones as rotation) or control group (given the modules after the control) for a total of 60 treated and 60 untreated foot rotation ends). Students completed a pre-test two weeks pads. Dermoscent BIO BALM was applied before and before their rotation start date and a post-test after after daily running (15–50 km) during 15 consecutive their rotation end date. A questionnaire was developed days and erythema/abrasion was evaluated on all foot to assess students’ learning experience using adaptive pads twice daily, before and after training, on days (D) learning modules. There was a statistically significant 1, D2, D4, D5, D8, D9, D12, D13 and D15. When difference (P = 0.019) in the post-test scores between bilateral lesions occurred on a dog on the same day students in the treatment and control groups. Students (BL), the time for healing was compared between trea- in both groups received a significant increase from pre- ted and untreated pads (chi-squared test). Erythema/ test to post-test (P < 0.001 for treatment group, abrasion was detected on 16/60 treated pads and 19/60 P = 0.002 for control group). Students’ views regarding untreated pads among which (respectively) 93.7% (15/ the adaptive learning modules was mostly positive. This 16) and 42.1% (8/19) were healed (P < 0.0015). Among study shows that the adaptive learning platform is an 15 BL dogs, 14/15 (93.3%) treated pads healed much effective novel method to augment clinical teaching in faster than the nontreated ones. More than half of trea- veterinary dermatology. This study also indicates that ted pads (8/15, 53.3%) were healed within one day of students perceive the use of adaptive learning modules application versus 0 for untreated pads (P < 0.001). as being beneficial for their education. After D4, 11/15 (73.3%) of treated feet healed versus 6/ Source of funding: Self-funded. 15 (40%) of untreated feet (P < 0.067). Dermoscent Conflict of interest: None declared. BIO BALM appears to be effective in aiding and

P-103 the first time in the dog. Two geriatric, male French bulldogs with pituitary-dependent hyperadrenocorticism First report of an epidermolytic foot pad and on trilostane treatment were presented with bilat- hyperkeratosis in a dog eral symmetrical alopecia and calcinosis cutis. More- R. GARCIA-FONTICOBA*,†, L. ORDEIX*,†,M. over, multiple mildly pruritic, firm, reddish and tightly CABRE* ,† and L. FERRER*,† adherent crusting papules were observed on the flank. *Servei de Dermatologia, Fundacio Hospital Clınic The removal of this protruding material gave rise to Veterinari, Universitat Autonoma de Barcelona, deep ulcers. Histopathological results (haematoxylin & Barcelona, Spain; †Departament de Medicina i Cirurgia eosin and Masson’s trichrome stains) showed transepi- Animals, Universitat Autonoma de Barcelona, Barcelona, dermal extrusion of degenerated collagen fibres perpen- Spain dicularly orientated and associated with an intense neutrophilic inflammatory infiltrate, crusting and necro- Hereditary foot pad hyperkeratoses (HFH) represent a tic debris. The histopathological findings were compati- group of canine genodermatoses most commonly ble with APD. The cause of these lesions remain described in the dogue de Bordeaux, Irish terrier and unknown. In one case, trilostane administration was Kromfohrlander.€ They are defined histologically as discontinued and the lesions resolved spontaneously nonepidermolytic hyperkeratosis. Mutations in the after six weeks. The other dog was euthanized four KRT16 (dogue de Bordeaux) and FAM83G (Irish ter- months after the diagnosis owing to seizures associated rier, Kromfhorlander)€ genes have been associated with with the growth of the pituitary adenoma and rapid the disease. We report the first case of epidermolytic worsening of neurological signs. At the time of eutha- foot pad hyperkeratosis in a dog. A 12-year-old neu- nasia the lesions were still present and had improved to tered male, mixed-breed dog was presented with a his- some extent. To the best of the authors’ knowledge, tory of severe painful foot pad hyperkeratosis from six this is the first report of canine APD. months old. Thick papilliform projections and fissures Source of funding: Self-funded. were observed in all foot pads. Histopathological exam- Conflict of interest: None declared. ination revealed severe papillated epidermal hyperplasia with orthokeratotic hyperkeratosis, intense hypergranu- losis, focal dyskeratosis and epidermolysis. The FAM83G and KRT16 gene mutations reported to be associated with this disease were not detected in this P-105 case. Human epidermolytic palmoplantar hyperkerato- Use of topic growth factors and minoxidil sis has been associated with mutation of the gene cod- sulfate in a female dog with black hair ing for keratin 9 (KRT 9). We fully sequenced this gene follicular dysplasia and found six single nucleotide polymorphisms (SNPs), † ‡ four of which cause amino acid change. However, none K. GAVAZZONI*, L. MARRA , E. FERRARINI § of these SNPs seemed to explain sufficiently the pheno- and V. KALTBACH † type of the dog. This is the first report of epidermolytic *Veterinary Clinic Santa Lucia, RS, Brazil; Small foot pad hyperkeratosis in a dog. Mutations in KRT16, Animal Clinic and Surgery, Universidade Federal de ‡ FAM83G or KRT9 genes do not seem to be the cause Goias, GO, Brazil; Pharmacy Farmec, Caxias do Sul, § of this phenotype. RS, Brazil; Pharmacy Bella Salute, Flores da Cunha, Source of funding: Self-funded. RS, Brazil Conflict of interest: None declared. Canine black hair follicular dysplasia (BHFD) is a rare disorder limited to the black coat regions affecting bicolour or tricolour dogs during the first few weeks of life. This case report describes the use of growth factors P-104 as a treatment modality for canine BHFD. A 5-year- Acquired perforating dermatosis in two French old, spayed, mixed-breed dog developed alopecia at eight months of age. The dog was otherwise healthy. bulldogs Histopathological evaluation of affected areas showed R. GARCIA-FONTICOBA*,†, P. J. SANCHO‡,D. follicular dysplasia and clumps of melanine in the hair FONDEVILA†, T. LIMA*,†, L. FERRER*,† and L. follicle canal and peri-follicular area compatible with ORDEIX*,† BHFD. The dog was unsuccessfully treated with mela- *Servei de Dermatologia, Fundacio Hospital Clınic tonin at 10 mg/day orally for three years. Five years Veterinari, Universitat Autonoma de Barcelona, after the diagnosis the dog had complete dorsal alope- Barcelona, Spain; †Department of Animal Medicine and cia and dry skin. Trichography was performed and Surgery, Universitat Autonoma de Barcelona, Barcelona, showed macromelanoses obscuring the normal hair Spain; ‡Clınica Veterinaria Dres. Sancho. Servicio de shaft architecture supportive of BHFD. A compounded Dermatologıa Veterinaria Dermasantboi. Sant Boi de moisturizing shampoo containing 2% colloidal oatmeal, Llobregat, Barcelona, Spain 2% hydroviton, 2% D-panthenol, 2% aloe vera, 2% Acquired perforating dermatosis (APD) is a group of Jaborandi tincture, 1% lactic acid and 0.2% pepper- human skin disorders characterized by collagen extru- mint essential oil was prescribed every seven days. In sion from the dermis. The pathogenesis is poorly under- addition, a compounded spray formulation containing stood and it is commonly associated with systemic 3% basic fibroblast growth factor (BFGF), 3% nano diseases. A similar clinical and histopathological condi- copper peptide and 5% minoxidyl sulfate was applied tion has been reported in cats. We describe APD for once daily to the dorsal region. During 40 days of the

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 98 Abstracts treatment, hair growth was noted covering the previ- P-107 ously alopecic areas. The haircoat was less dense and had a different quality. Maintenance therapy with the Idiopathic linear pustular acantholytic compounded formulations included weekly baths and dermatosis in a border collie three times a week spray application. In conclusion, the T. NUNES RODRIGUES and S. VANDENABEELE combination of BFGF, nano copper peptide and Small Animal Department, Faculty of Veterinary minoxidil should be considered as a treatment option Medicine, Ghent University, Merelbeke, Belgium for canine BHFD. Source of funding: Self-funded. Idiopathic linear pustular acantholytic dermatosis Conflict of interest: None declared. (ILPAD) is a very rare condition with an unknown aetiology or predisposition. A 10-month-old, female spayed border collie was presented with a six week history of a pruritic linear raised solitary lesion. The lesion P-106 involved the fifth pad and partially the metacarpal pad, and extended midway between the carpal pad and the Nodular dermatofibrosis and cystic renal olecranon of the left forelimb. It consisted of coalescent disease in a beagle without FLCN mutation: a crusts. The affected paw pads presented with moderate case report erythema and erosion. Previous treatments with short courses of oral antibiotics, antifungals, anti-inflamma- B. RIVERA*, V. BALAZS*, C. TORO† and S. ‡ tories and glucocorticoids did not improve the lesions. STUBBS Cytological examination of impression smears of the *Oftaderm, Clınica especializada en Oftalmologıa y lesion revealed high numbers of Malassezia yeast. Two † Dermatologıa Veterinaria, Santiago, Chile; Department punch biopsies were taken for histopathological exami- of Veterinary Medicine, University of Cambridge, UK; ‡ nation. Topical treatment with orbifloxacin, mometa- London School of Hygiene and Tropical Diseases, sone furoate monohydrate and posaconazole (Posatex, London, UK Intervet; Boxmeer, the Netherlands) and oral ketocona- Nodular dermatofibrosis (ND) is a rare syndrome ini- zole at 5 mg/kg/day was prescribed until the biopsy tially described in German shepherd dogs. The disease results became available. Histopathological results is characterized by the presence of multiple dermal and showed neutrophilic mural and luminal folliculitis with subcutaneous nodules primarily affecting the legs, head pronounced acantholytic cells, and epidermal exocytosis and ears. It also is associated with polycystic kidneys, of neutrophils. There was a significant improvement renal cystadenomas or cystadenocarcinomas, and less with topical medication after two weeks. Ketoconazole commonly with uterine tumours. This condition has was discontinued and oral prednisolone at 0.7 mg/kg been linked to an autosomal dominant mode of inher- once a day was prescribed at the time of histopatholog- ence and to mutations of the gene encoding folliculin ical results. The lesions resolved after two weeks of (FLCN). An 8-year-old, intact male beagle presented therapy. The prednisolone was weaned-off and topical with several well-circumscribed skin nodules located on treatment discontinued. The case remained controlled the extremities, head and pinna that progressed over a for nearly three years with occasional use of topical period of two years. No other clinical signs were pre- therapy and short courses of prednisolone. In a similar sent and haematological as well as biochemical parame- way to the previous cases reported in the literature, ters were unremarkable. Abdominal ultrasound showed ILPAD affects only one area of the body and responds bilateral cortical renal cysts suggesting a degenerative/ very quickly to glucocorticoids. neo-proliferative inflammatory process. One of the cysts Source of funding: Self-funded. in the left kidney was large and it did not appear to Conflict of interest: None declared. affect the renal parenchyma significantly. A PCR blood test did not reveal germline mutations in the FLCN gene. These findings correlate with other reported cases of ND. This dog has lived for two years since diagno- P-108 sis, exceeding by far the life expectancy found in cases of malignant renal cysts, suggesting the absence of neo- P-1A case of ulcerative neutrophilic dermatitis plasia. It is the first time that this disorder has been resembling human bromoderma in a dog described in beagles. The fact that FLCN mutations C. DEDOLA*, A. GALLUCCI†, F. IBBA‡,F. were not detected suggests that alterations in different ENNAS§, I. PORCELLATO¶ and C. genes may be involved in this particular form of ND. BRACHELENTE¶ Future studies are still required to determine the aetiol- *Centro Veterinario Ichnos, Cagliari, Italy; †Veterinary ogy of this FLCN-independent nonmalignant ND, now Centre La Fenice - Neurology Section, Cagliari, Italy; described in beagles. ‡Animal House Clinic, Capoterra, Italy; §Karel Source of funding: Self-funded. Veterinary Clinic, Cagliari, Italy; ¶Department of Conflict of interest: None declared Veterinary Medicine, University of Perugia, Perugia, Italy Bromoderma is a cutaneous adverse reaction described in humans and caused by ingestion, inhalation or direct contact with bromides. Lesions are characterized by pustular and acneiform eruptions as well as nodules and ulcers affecting the face, scalp and limbs. A 2-year- old, female Maremma sheepdog was referred for the presence of six months duration painful ulcers with

© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD, Veterinary Dermatology, 31 (Suppl. 1), 6–109 Abstracts 99 violaceous borders and multiple dermal and subcuta- islets. The pituitary tumour showed a loss of normal neous haemorrhagic nodules on the bridge of the nose, reticulin stromal pattern and multifocal periodic acid the dorsal aspect of the front paws and on all four legs. Schiff-positive neoplastic cells. Mitoses were six per 10 Lesions had not responded to antibacterial and HPF. The tumour was PIT1-negative, TPIT- and immunomodulatory therapy. Nine months before, the ACTH-positive. P53 was negative and Ki67 index was dog had been diagnosed with suspected idiopathic epi- 7%. According to human classifications, this was a lepsy and treated with potassium bromide (KBr) ever pituitary neuroendocrine tumour (PitNET) grade 1b, a since. Histopathological examination of lesions revealed corticotroph adenoma with high Ki67 labelling index an interstitial neutrophilic dermatitis extending multifo- (based on the WHO 2017 classification). This tumour cally to the subcutaneous tissue. All special stains were caused cortical hyperadrenocorticism, skin fragility and negative for infectious agents, and owing to the lack of diabetes mellitus. tropism for follicular structures as well as negative bac- Source of funding: Self-funded. terial and fungal cultures, a diagnosis of sterile neu- Conflict of interest: None declared. trophilic process similar to pyoderma gangrenosum was made. A cutaneous drug reaction to KBr was suspected, because it was the only continuous drug in the dog’s history before the first appearance of lesions. P-110 Potassium bromide was discontinued and after six weeks the ulcerative lesions were completely healed. Alopecia and suprasellar secreting neuro- The present report describes a case of an ulcerative endocrine tumour in a dog neutrophilic dermatitis associated with KBr administra- C. GIMENEZ-ACOSTA*, B. HUBERT*, K. tion in an epileptic dog, resembling human bromod- MATIASEK†, L. BLOND*, C. BAUDIN*, F. erma. Corticosteroid treatment was ineffective in DEGORCE-RUBIALES‡ and S. PIAZZA* controlling skin lesions and only drug withdrawal led *Veterinary Hospital Medicine Languedocia, to clinical resolution. Montpellier, France; †Clinical & Comparative Source of funding: Self-funded. Neuropathology Laboratory, Ludwig-Maximilians- Conflict of interest: None declared. University, Munich, Germany; ‡Laboratoire d’Anatomie Pathologique du Sud-Ouest, Toulouse, France Most of the hormone-active adenomas secrete prolactin (50%) and more rarely somatotropin (10%) or corti- P-109 cotropin (5%). Secretion from another tumour type, Skin fragility in a cat with an ACTH- especially in case of suprasellar extension, has not been producing pituitary neuroendocrine tumour reported previously. A 9-year-old, intact female bearded (PitNET) collie was presented with a major alopecia, especially involving the trunk, neck and tail, and sparing the B. BRUNETTI*, L. V. MUSCATELLO*, F. head. The dog was diagnosed with hypothyroidism and † ‡ § CLEMENTE , S. ASIOLI , B. BACCI and F. treatment with levothyroxine was unsuccessful. An § ABRAMO adrenocorticotropic hormone stimulation test was con- *Department of Veterinary Medical Sciences, University sistent with increased adrenal gland response suggestive † of Bologna, Italy; Associate Veterinary Clinic “San of hyperadrenocorticism. No clinical improvement was ‡ Luca”, Bologna, Italy; Department of Biomedical and visible after oral trilostane treatment (1 mg/kg daily for Neuromotor Sciences, University of Bologna, Italy; 30 days). Microscopic examination of skin biopsies § Department of Veterinary Sciences, University of Pisa, revealed a severe atrophic dermatosis, lack of inflam- Italy mation, an extremely thin dermis, marked orthokera- Feline acquired skin fragility syndrome is a rare condi- totic hyperkeratosis, follicular plugging, empty tion with multiple underlying causes, including sponta- follicular ostia, atrophic sebaceous and apocrine glands, neous and iatrogenic hyperglucocorticism and diabetes and the preponderance of telogen and kenogen follicles. mellitus amongst others. A 14-year-old, female Euro- An endocrinopathy was suspected. High-dose dexam- pean cat was presented for severe sudden and diffuse ethasone suppression testing was consistent with an skin lacerations, either spontaneous or induced by adrenal secreting neoplasia or pituitary-dependent minor trauma. The cat had a two year history of glyco- hyperadrenocorticism. Abdominal ultrasound revealed suria and untreated polydipsia and polyuria. On clinical mild symmetrical increase of adrenal gland size and examination, it had poor physical condition, dehydra- hepatomegaly. The clinical profile of this patient was tion, severe plantigrade stance, thin, fragile skin, and ambiguous with no polyuria-polydipsia, no cutaneous lacerations without skin hyperextensibility. The suspi- or muscular calcinosis and normal alkaline phosphatase cion was of acquired skin fragility, and owing to the plasmatic activity. The dog developed neurological clin- poor condition of the patient and severity of skin dam- ical signs and was euthanized. A pituitary mass was age, compassionate euthanasia was performed. At confirmed during necropsy as a suprasellar neuroen- necropsy, the adrenal glands were both enlarged and docrine-type cell tumour from pars distalis and the the pituitary gland had a tumour measuring adrenal glands were slightly hyperplastic. This case 10 x 10 x 12 mm. Histopathological examination of the report describes an unusual form of hyperadrenocorti- skin revealed a very thin epidermis with a single layer cism in a dog. of keratinocytes, atrophic dermis and sparse and thin Source of funding: Self-funded. collagen. Transmission electron microscopic investiga- Conflict of interest: None declared. tion showed no structural alterations of the collagen. Both the adrenal glands had cortical hyperplasia. In the pancreas, there was amyloidosis of the Langerhans

P-111 enlarged and had a moderately thick wall. The other uterine horn had a thin wall, and was expanded, filled Skin necrosis in a cat likely caused by with purulent brown liquid. The histopathological subcutaneous injection of enrofloxacin results of the neoplastic testicle revealed a parenchyma A. MIYA*, E. HIRATA†, N. MURAYAMA‡ and Y. formed by many tubules, separated by a very dense OSHIMA-TERADA‡ stroma. These had elongated and palisading cells, the *Okusawa Subaru Animal Clinic, Setagaya, Tokyo, majority with multiple nucleoli. The other testicle was Japan; †Misaki Animal Hospital, Suginami, Tokyo, hypoplastic. In relation to the uterus, the largest horn Japan; ‡Dermatology Services for Dogs and Cats, Koto, had a reduced mucosa and submucosa, with infiltration Tokyo, Japan of active neutrophils and macrophages, a thin muscular layer, marked congestion and angiogenesis. The horn Although there is one case report on injection site that had the neoplastic testicle attached showed evi- necrosis due to enrofloxacin, no detail was provided. dence of endometrial cystic hyperplasia along with The aim of this study was to report the histological inflammation. In conclusion, this is the first report in examination of the injection site after subcutaneous Peru of persistent Mullerian€ duct syndrome with hyper- enrofloxacin caused injection site necrosis in a cat. A 2- oestrogenism, sertoli cell tumour, endometrial cystic year-old, neutered mixed-breed cat presented for ulcera- hyperplasia and pyometra in a male schnauzer. tions over the dorsal neck. The cat had been given an Source of funding: Self-funded. injection of enrofloxacin (5 mg/kg subcutaneously once Conflict of interest: None declared. daily, Baytril 2.5% Injectable Solution, Bayer, Yaku- hin, Ltd.; Osaka, Japan) and prednisolone 1–2 mg/kg s.c. once daily (Prednisolone Injection Solution KS, Kyoritsu Seiyaku Corp.; Tokyo, Japan) in the dorsal neck over the course of two months as a treatment of P-113 anaemia. Three weeks after these treatments, a skin Post-clipping hair follicle arrest and lesion of 1 cm full-thickness ulceration was observed. glucocorticoid-induced alopecia treated with However, at the time of presentation, ulceration malt extracts made in presence of Rhizopus (8 9 5 cm) had spread over the dorsal neck. Our histological examination revealed full-thickness coagulation oryzae in dogs necrosis of the epidermis, dermis and adnexa with Y. NAKAMURA* and T. FUKASE† extension into the panniculus. The lesion was almost *Katsuragi Institute of Life Sciences, Tokyo, Japan; completely resolved after nine months of simply rinsing †Department of Veterinary Medicine, Faculty of the affected area with normal saline solution and Veterinary Medicine, Okayama University of Science, changing the moist wound dressing every day. Based Imabari, Japan on the medical history of and histological examination In the treatment of one dog with post-clipping hair fol- on this cat, we have concluded that the high pH of licle arrest and one dog with glucocorticoid-induced enrofloxacin (approximately 11) may have caused the alopecia, we evaluated the efficacy of a product con- full-thickness coagulation necrosis in the cat. taining water extracts of the malt made in the presence Source of funding: Self-funded. of Rhizopus oryzae (R&U 30, Ushikoshi Research Insti- Conflict of interest: None declared. tute for Physiology; Chiba, Japan). This product is known to develop a regulatory effect on steroid hor- mones. The first case was a 13-year-old, castrated long- haired miniature dachshund, with body weight 6.7 kg, P-112 showing alopecia of the flanks and ventrum post-clip- Hyperoestrogenism, sertoli cell tumour and ping. One tablet (120 mg) of the product was adminis- pyometra in a male schnauzer with persistent tered orally twice a day. Hair regrowth occurred by 40 € days after starting the medication. The second case was Mullerian duct syndrome a 13-year-old, castrated toy poodle, with body weight † G. MONTOYA*, E. ESCOBEDO*, S. PIZARRO ,G. 5.0 kg, showing alopecia in the right flank resulting SALAS† and R. RAMIREZ† from topical glucocorticoid use over three months. One *Department of Dermatology, Scientific University of tablet of the tested product was administered orally the South, School of Veterinary Medicine, Lima, Peru; once a day. After 28 days, the hair began to regrow, † Department of Pathology, Scientific University of the with the colour of the newly growing hair changing to South, School of Veterinary Medicine, Lima, Peru brown instead of cream. The hair regrowth in this case Persistent Mullerian€ duct syndrome is a rare form of is thought to have been achieved earlier than in the pseudohermaphroditism in male dogs, mainly described case without treatment after stopping a topical gluco- in toy schnauzers. This report describes a case with this corticoid. The tested product may be helpful for some condition. A 10-year-old male schnauzer with gynaeco- kinds of canine alopecia. mastia, comedones, linear erythema of the preputial Source of funding: Self-funded. skin, and only one descended testicle was sent for an Conflict of interest: None declared. ultrasound examination, revealing the other testicle to be abdominal and enlarged to 10 cm. Needle aspiration and cytological evaluation were suggestive of a Sertoli cell tumour and so the dog was castrated. During surgery, it was observed that the neoplastic testicle was nodular, whitish coloured with dark red areas and was cranially attached to a uterine horn that was slightly

P-114 noted followed by a relapse and worsening of the lesions three months later. This observation suggests Sensory neuropathy (acral mutilation the likelihood of a genetic component in the develop- syndrome) in three miniature pinschers ment of this disease as both the dog and its clone had ,† ‡ ,† developed the disease at the same age regardless of L. ORDEIX* , T. LIMA and L. FERRER* *Servei de Dermatologia, Fundacio Hospital Clınic their living conditions. Moreover, neutering, despite allowing an identical temporary improvement, was not Veterinari, Universitat Autonoma de Barcelona, Barcelona, Spain; †Departament de Medicina i Cirurgia more effective when performed earlier than later. Animals, Universitat Autonoma de Barcelona, Barcelona, Source of funding: Self-funded. Spain; ‡Hospital Veterinari Canis, Girona, Spain Conflict of interest: None declared. Hereditary sensory neuropathies are uncommonly diagnosed in dogs and usually reported as “acral mutilation syndromes” (AMS). Similar to humans, genetic causes P-116 have been described. A point mutation in a lincRNA upstream of the GDNF gene has been associated with Conformational abnormality as a frequent AMS in sporting dog breeds (pointers, English springer cause of pododermatitis in canines spaniel, French spaniel). Although AMS has been A. TIWARI, B. KHAJURIA, V. VERMA and K. described previously in one miniature pinscher, at this KANT time it is not known if the disease also has a genetic Suvidha Vet Care and Research Centre, Jammu & origin in this breed. The present report describes three Kashmir, India additional cases of AMS in miniature pinschers. Three unrelated dogs, two females and one male miniature Canine pododermatitis is a common clinical presenta- pinscher were presented with intense licking and biting tion that causes paw licking or lameness. This study of the hind limbs leading to severe self-mutilation of was conducted to analyse the causes of pododermatitis several claws, digits and foot pads. The age of disease in three breeds of dogs. Labrador retrievers (labs), bea- onset was between four and 15 months. Neurological gles and golden retrievers (n = 15 of each breed) over 1 examination confirmed acral analgesia of the hind feet year of age with pododermatitis were recruited for this in all cases. Mutation of the lincRNA upstream of the study. All dogs had hair plucks, skin scraping and cyto- GDNF gene was not found. One case was lost to fol- logical evaluation performed. Biopsies were taken from low-up after diagnosis, while the other two showed par- eight labs and six beagles, bacterial culture and sensitiv- tial response to gabapentin and fluoxetine. The ity tests were performed in 10 labs, 12 beagles and six miniature pinscher seems to be one of the breeds where golden retrievers with deep pyoderma. The cause of this specific neuropathy occurs, although it is not asso- pododermatitis was found to be demodicosis in four ciated with the mutation described for other breeds. labs, four beagles and two golden retrievers, Malassezia Therefore, this breed may constitute an interesting spp. in three labs, two beagles and four golden retriev- model for the identification of new gene mutations ers, atopic dermatitis in five labs, six beagles and six causing AMS in dogs and potentially for further insight golden retrievers, orthopaedic pain in three labs, one into human neuropathies as well. beagle and three golden retrievers, and a conforma- Source of funding: Self-funded. tional defect of the foot in five labs and nine beagles. Conflict of interest: None declared. This conformational defect caused ventrally fused pads to form a horseshoe bridge, which leads to weight bear- ing on interdigital skin. Cytological results of cases with the conformational defect revealed degenerate neu- P-115 trophils and macrophages, and biopsy showed no neoplastic growth. Increased body weight with splayed feet Cloned alopecia X in a Pomeranian dog predisposes to this painful condition, causing inflamma- S. DELEPORTE*, P. PRELAUD† and C. LAFFORT‡ tion of hair follicles and results in chronic inflammation *Dermatology department, Lorrainevet, Ludres, France; in the feet. Conformational pododermatitis was most †Dermatology department, Advetia Centre Hospitalier commonly seen in labs and beagles, and these patients Vet eriniare, Velizy, France; ‡Dermatology department, were the most difficult to treat. Cases of conforma- Clinique Vet erinaire Alliance, Bordeaux, France tional pododermatitis are rarely cured and require long-term management. Alopecia X is a frequent cause of alopecia in Pomera- Source of funding: Self-funded. nian dogs. The pathomechanism is still obscure and a Conflict of interest: None declared. genetic basis suspected. This report of a cloned Pomeranian argues strongly for a genetic basis for this condition. A 2-year-old male Pomeranian was presented early 2014 with alopecia X that had been progressing over a year. It was neutered in April 2014 and hair regrowth was observed and quickly followed by a relapse of the hair cycle arrest. The patient died acci- dentally in September 2014. His owner immediately arranged for his dog to be cloned in South Korea (Sooam Biotech; Seoul, South Korea). A viable clone was born in August 2015 and was returned to its owner in France in March 2016. It also presented with the first clinical signs of alopecia X at one year of age and was immediately neutered. Partial hair regrowth was

P-117 P-118 A case report of sterile granuloma/ pyogranuloma syndrome in a cat Junctional epidermolysis bullosa in a domestic cat K. MORI*, Y. OSHIMA-TERADA*, H. KONDO† K. SHIBATA*, K. OKADA†, Y. KAGAWA† and M. and N. MURAYAMA* ‡ *Dermatology Services for Dogs and Cats, Tokyo, TAKANOSU † *DVMs Animal Medical Center Yokohama, Kanagawa, Japan; Nihon University College of Bioresource † ‡ Sciences, Kanagawa, Japan Japan; North Lab, Hokkaido, Japan; Nasunogahara Animal Clinic, Tochigi, Japan Sterile granuloma/pyogranuloma syndrome (SGPS) is a very rare disease in cats. The pathogenesis and treat- Junctional epidermolysis bullosa (JEB) is a disorder ment have not been fully evaluated. This case report characterized by subepidermal blistering at the lamina describes SGPS in a cat and the efficacy of essential lucida. In humans, mutations of LAMA3, LAMB3, fatty acids (EFA) for this disease. A 3-year-old, cas- LAMC2, COL17A1 and ITGB4 have been identified as trated male American short hair cat, kept indoors, pre- responsible for JEB. Our objective was to examine the sented with alopecic nodules on the nasal bridge cause of JEB in an 11-month-old, female mixed-breed without pruritus. The physical examination revealed no cat referred for recurrent ulcerative dermatitis. Clinical abnormalities. Trichogram, skin scrapings and impres- examination revealed ulcerations and erosions associ- sion smears were unremarkable. Results of complete ated with crusts on the pinna, head and axilla. In addi- blood cell count and serum chemistry analysis were tion, erosions with scales of multiple foot pads and within normal limits. The histopathological findings onychomadesis of all claws were present. The cat also revealed pyogranulomatous dermatitis consisting of had severe stomatitis. Histopathological examination neutrophils and macrophages extending from the der- revealed subepidermal bullae. Periodic acid Schiff stain mis to the subcutaneous layers. Foreign bodies, infec- and immunostaining for collagen type IV were positive tious agents and atypical cells were not detected by on the bottom of the bullae, indicating that the dermo- periodic acid Schiff, Ziehl–Neelsen, Giemsa and tolu- epidermal separation occurred at the lamina lucida. idine blue staining. Based on the clinical and These clinical and histopathological findings were con- histopathological findings, the cat was diagnosed with sisted with JEB. Real-time PCR of skin cDNA revealed SGPS. Two capsules once daily of EFA (Antinol, V abnormal splicing events in exons 22–26 of LAMA3. and P; Tokyo, Japan) were administered and the lesions Although the whole genome sequencing of the cat improved after one month. However, two months after revealed no mutations in open reading frames of the initial treatment, the alopecic nodules relapsed. The LAMA3, LAMB3, LANC2 and ITGB4, mutations were cat was treated with two capsules once daily of another identified in eight of 191 splicing factors that may have EFA (Moegicap, Veterinarian Medical Development; induced aberrant splicing variants of laminin 332. These Saitama, Japan) containing higher concentrations of findings suggest that a partial defect in the laminin 332 omega-3 fatty acids. The lesions resolved one month a3 chain might be the cause of JEB in this cat. Further later and did not relapse for one year after the initial study is need to demonstrate if mutated splicing factors visit. To the best of our knowledge, this is the first caused the aberrant splicing variants of laminin 332 in report of SGPS in a cat treated with EFA. This report this case. indicates that omega-3 fatty acids, which have anti- Source of funding: Self-funded. inflammatory effects, could be an effective treatment Conflict of interest: None declared. for SGPS. Source of funding: Self-funded. Conflict of interest: None declared.

FC-66 Restrictive diet and dietary challenge guided FC-67 by prick test in the diagnosis of food allergy in Evaluation of clinical signs related to adverse dogs with chronic pruritus associated with food reaction (AFR) in dogs using a new atopic dermatitis behaviour-recognition approach C. ALCALA, J. POSSEBOM, V. GMYTERCO and S. M. WERNIMONT*, J. L. WEEMHOFF*, J. M. † M. R. FARIAS MACLEAY*, J. J. BREJDA and K. L. GROSS* † Escola de Ciencias^ da Vida, Pontifıcia Universidade *Hill’s Pet Nutrition, Inc. Topeka, KS, USA; Alpha Catolica do Parana, PUCPR, Curitiba, Brazil Statistical Consulting, Lincoln, NE, USA The aim of this study was to evaluate the prick test Quantifying intervention response in canine adverse with extracts of food proteins in the diagnosis of food food reactions (AFR) is a challenge. A device providing allergy in dogs with atopic dermatitis. To this end, 34 quantitative scratching, shaking, resting and sleep data dogs (Group 1) with atopic dermatitis syndrome and 10 could assist management. We paired this innovative healthy dogs (Group 2) were screened, which were sub- tool with the COSCAD’18 and examined behaviours jected to a prick test with extracts of bovine protein, over time in an IACUC-approved prospective study. chicken, egg, pork, fish, milk, soy and wheat and envi- Forty-seven dogs diagnosed by food elimination trial at ronmental allergens, considering positive papules larger general veterinary practices and consuming a novel or than 3 mm from the negative control. Dogs with a pos- hydrolysed protein food provided complete data. Dogs itive food test received a homemade diet based on their wore a collar-mounted activity monitor and veterinary result, PVAS and CADESI-4 being evaluated on days CADLI scores and owner PVAS scores were performed 0, 30, 60 and those who improved were challenged with on day (D) and D21. Machine learning-based beha- positive protein for 7 days. In Group 2, one dog viour recognition algorithms quantified the total dura- reacted to wheat and another to milk. In Group 1, four tion of scratching (s/day), shaking (s/day), resting (h/ (12%) reacted to environmental allergens, four (12%) day) and sleeping (h/day); sleep quality was evaluated to food and 26 (76%) to both, 30 of which were started with an algorithm based on the absence of night-time on diet trials and 25 of 30 remained in the study. Per- disturbance (0–100: highly disturbed-undisturbed sleep). centage improvement of PVAS (P = 0.0001) and Behaviours, CADLI and PVAS scores were analyzed in CADESI-4 (P = 0.0001) between days 0 and 60 was SAS using linear mixed-models with diet, time, and observed in 21 (84%) dogs, with no difference between DietxTime as fixed-effects and significance P < 0.05. days 30 and 60, and all increased PVAS (P = 0.0001) Duration of behaviours (mean Æ SD) were: scratching: and CADESI-4 (P = 0.039) after the challenge. Bovine, 108 Æ 136 s/day; shaking: 39.7 Æ 27.6 s/day; resting: wheat and chicken proteins were the most reactive. A 9.1 Æ 2.0 h/day; sleeping: 12.6 Æ 2.2 h/day; and sleep conclusion is that dogs with atopic dermatitis are gener- quality: 90.1 Æ 8.4. There were no statistically signifi- ally polysensitized and the prick test can serve as a cant differences between the CADLI and PVAS scores guide for establishing dietary protocols and assessing collected at study entry and day 21 (CADLI: 4.43/50 food allergy in them. vs. 3.65/50, P = 0.09; PVAS: 3.69/10 vs. 3.49/10, Source of funding: Coordenacßao~ de Aperfeicßoamento de P = 0.50). Quantitative, objective behaviour data from Pessoal de Nıvel Superior (CAPES). wearable activity monitors complements the COSCAD Conflict of Interest: None declared. ‘18 and is free of recall bias. Quantitative behaviour data provides a new reference for clinical studies of dogs with AFR. Source of Funding: Hill’s Pet Nutrition. Conflict of Interest: Authors are employees of or were compensated (JJB) by Hill’s Pet Nutrition, Inc.

FC-68 FC-69 Quantification of behaviours in dogs with Successful management of clinical signs related atopic dermatitis with a machine learning- to adverse food reaction in dogs using a food based approach with a new approach J. L. WEEMHOFF*, S. M. WERNIMONT*, J. M. J. M. MACLEAY*, J. L. WEEMHOFF*, S. M. MACLEAY*, J. J. BREJDA† and K. L. GROSS* WERNIMONT*, J. J. BREJDA† and K. L. GROSS* *Hill’s Pet Nutrition, Inc. Topeka, KS, USA; †Alpha *Hill’s Pet Nutrition, Inc. Topeka, KS, USA; †Alpha Statistical Consulting, Lincoln, NE, USA Statistical Consulting, Lincoln, NE, USA Quantifying intervention response in canine atopic der- Quantifying intervention response in canine adverse matitis (CAD) is a challenge. A device providing quan- food reaction (AFR) is a challenge. Devices providing titative scratching, shaking, resting and sleep data quantitative scratching, shaking, resting and sleep data could assist management. We paired this tool with the could assist management. We paired this tool with the COSCAD’18 and examined behaviours in an IACUC- COSCAD’18 in an IACUC-approved randomized, con- approved prospective study. Dogs with CAD and vet- trolled, double-masked prospective study. Dogs diag- erinarian-assessed CADLI scores ≥8/50 and an owner- nosed with AFR, consuming a novel or hydrolysed reported PVAS scores ≥3.5/10 were enrolled and wore protein food were enrolled. CADLI and PVAS were an activity monitor. Machine learning algorithms quan- recorded on day (D)0, D21 and D42 and dogs wore tified the total duration of dermatitis-related behaviours activity monitors throughout. Dogs consumed their per day: scratching (s/day), shaking (s/day), resting (h/ food (0–21 days) and then were randomized to test day) and sleeping (h/day). Ninety-five dogs had com- (TF) or control (CF) food for 22–42 days. The TF con- plete data. Mean Æ SD CADLI (13.8 Æ 0.78) and tained egg, n3-6-9 fatty acids, and the CF hydrolysed PVAS (6.86 Æ 3.6) scores confirmed clinical signs con- animal protein. Duration of scratching (s/day), shaking sistent with entry criteria. Activity data reported as (s/day), resting (h/day), sleeping (h/day), and sleep mean Æ SD (median, interquartile range (IQR): quality (0–100:highly disturbed-undisturbed sleep). scratching: 254 Æ 319.1 (131.0, 210.0) s/day, shaking: Analysis was linear mixed-models with diet, time, and 44.1 Æ 31.7 (35.5, 32.5) s/day, resting: 9.0 Æ 1.8 (9.0, diet 9 time as fixed-effects and a significance threshold 1.8) h/day, and sleeping: 12.9 Æ 2.1 (13.0,2.3) h/day. of P < 0.05 in SAS. Twenty-three TF and 21 CF com- Scratching duration was greater and more variable pleted study. Baseline and D42 CADLI and PVAS though shaking duration was similar to each of two scores were similar (P > 0.16). In 35 dogs stratified on comparator groups: a group of pruritic dogs at general CADLI (> or <8/50) there were no differences between practice clinics (mean Æ SD scratching: 96.9 Æ 11.4 TF and CF (P > 0.60). Dogs beginning with CADLI and shaking: 49.2 Æ 5.5 s/day) and a large population score ≤8/50 remained in that category. In activity data of dogs with various health states (median, IQR from 32 dogs there were no differences between TF & scratching: 46, 80 and shaking: 31, 31). Quantitative, CF in change from baseline for any endpoint objective behaviour data from wearable activity moni- (P > 0.30), and no linear trends (P > 0.20). Pet owner tors complements the COSCAD ‘18 and is free of recall satisfaction was similarly positive between groups. bias. Quantitative behaviour data provides a new refer- Feeding a food formulated for AFR resulted in success- ence for clinical studies of dogs with CAD. ful transition and comparable control of clinical signs Source of Funding: Hill’s Pet Nutrition. vs. feeding a hydrolysed protein-based food. Conflict of Interest: Authors are employees of or were Source of Funding: Hill’s Pet Nutrition. compensated (JJB) by Hill’s Pet Nutrition, Inc. Conflict of Interest: Authors are employees of or were compensated (JJB) by Hill’s Pet Nutrition, Inc.

FC-70 Amongst non-inclusion criteria were other diseases, ges- tation or lactation, secondary infections, impossibility Diagnosis of feline cutaneous adverse food to stop flavored medications. Drug usage was allowed reactions with extensively hydrolysed protein only in precise circumstances e.g. needing oral or local diet antibiotics during the elimination phase. After inclusion U. MAYER*,1, K. BERGVAL†, L. BECO‡,J. visit V0, cats were fed the test diet (extensively hydrol- FONTAINE§, T. BORD¦, S. AHMAN**, R. ysed protein and purified corn starch) for eight weeks, ALMELA*, L. BOUTIGNY†† and I. LESPONNE†† with follow-up visits at day 30 (V1) and 60 (V2). Then *Kleintierspezialisten, Augsburg, Deutschland; cats were challenged with previous diet untill V3 (flare †Department of Clinical Sciences, University of up, 14 days otherwise), and back to test diet until study Agriculture, Uppsala, Sweden; ‡Cabinet vet erinaire completion. Thirty-four cats were enrolled, ten com- MonVt, Spa, Belgium; §Animal Clinic, Bruxelles, pleted both phases. Eight of these had lesions improved Belgium; ¶Clinique vet erinaire de l’Arche, Salon-de- with the test diet (SCORFAD decreasing from median Provence, France; **DjurAkuten AB, Stockholm, 3.5 [2;5] at visit (V)0 to 1.5 [0;6] at V2, owners mention- Sweden; ††Royal Canin, Aimargues, France ing skin improvement after 12 days in average). Two of them, also experiencing worsening during re-challenge, The diagnosis of cutaneous Adverse Food Reactions were diagnosed as cAFR cases. Owner-assessed fecal (cAFR) in pets relies on performing an elimination trial scores confirmed good digestive tolerance of the food. followed by re-challenge. The objective of this prospec- The results show that this food may constitute a valu- tive multicenter study was to assess the diagnostic effi- able tool for feline cAFR diagnosis. Further studies are cacy of an extensively hydrolysed protein-based food in warranted to confirm it. cats. Adult cats with suspected cAFR were included Source of Funding: Royal Canin SAS. into the study by dermatology specialists, if also meet- Conflict of Interest: IL is employee of Royal Canin SAS. ing criteria for pruritus (Pruritus Visual Analog Scale LB is a former employee of Royal Canin SAS. equal or greater to 1) and/or lesions (Scoring Feline Allergic Dermatitis (SCORFAD) equal or greater to 4).

FC-71 determine if there is association between resistance to first and second tier antibiotics and presence of mecA; Prevalence of methicillin resistant and 3) determine if there is an association between Staphylococcus pseudinteremedius in dogs with MRSP and previous antibiotic use. Sixty-eight clinical skin and ear infections in South Africa samples from five geographically dispersed laboratories C. PRIOR*, M. KARAMA†, A. MOODLEY‡ and in RSA were collected over a 24-month period. SP were A. LEISEWITZ* detected by a standard laboratory method and antimi- *Department of Companion Animal Clinical Studies, crobial susceptibility testing by disc diffusion. Possible Faculty of Veterinary Science, University of Pretoria, MRSP isolates were flagged when resistance to oxacillin Onderstepoort, South Africa; †Department of was observed. All isolates were confirmed as SP and Paraclinical Sciences, Veterinary Public Health, Faculty genotyped for the mecA gene by PCR. Fifty-three iso- of Veterinary Science, University of Pretoria, lates were confirmed to be SP (78%) and only 44 Onderstepoort, South Africa; ‡Department of Veterinary (83%) were MRSP. Thirty-two MRSP were isolated and Animal Science, Faculty of Health and Medical from pyoderma (32/44, 73%) and 8/44 from otitis Sciences, University of Copenhagen, Copenhagen, (18%). All mecA positive samples were recovered from Denmark; ‡International Livestock Research Institute, dogs that had been exposed to antimicrobials previ- CGIAR AMR Hub, Nairobi, Kenya ously (50/68, 73.5%). MRSP isolates are common among dogs with pyoderma and otitis in South Africa. Staphylococcus pseudintermedius (SP) is an important There is an urgent need for better laboratory diagnosis opportunistic pathogen, frequently associated with der- of MRSP and surveillance of dogs presenting pyoderma matitis and otitis in small animals. The emergence and and otitis in South Africa. Moreover, diligent antibiotic rapid expansion of methicillin resistance is problematic stewardship will be crucial to prevent a deterioration in due to multi-drug resistance and reduced treatment this situation in South Africa. options. We aimed to 1) confirm that standard labora- Source of funding: Zoetis Animal Health. tory methods used at five regional veterinary labs in Conflict of interest: None declared. South Africa (RSA) can accurately identify SP; 2)

FC-72 FC-73 Evaluation of owner conduct in the Topical corticosteroids and urea for the management of dogs with atopic dermatitis, in treatment of canine atopic dermatitis the municipality of Rio de Janeiro-RJ T. MATSUDA*, N. NUNOKAWA*, S. HIRANO* † M. SILVA* and M. SILVA-CARVALHO† and K. IYORI *Nunokawa Dogs and Cats Hospital, Kanagawa, Japan; *Estacio de Sa University (UNESA), Rio de Janeiro, † Brazil; †Federal University of Rio de Janeiro, Rio de Vet Derm Tokyo, Kanagawa, Japan Janerio, Brazil This study investigated the efficacy of topical corticos- Dermatological problems are currently one of the main teroids (TCSs) of varying potencies with and without reasons for going to veterinary clinics for small ani- topical moisturizer in dogs with atopic dermatitis (AD). mals. Canine Atopic Dermatitis is an inflammatory Seven dogs with chronic AD were enrolled. Three of skin disease generated by an exacerbated reaction of four digital skin lesions of each dog were randomly the immune system to different types of allergens. It is selected for treatment with one of three topical regi- a disease of genetic origin, in which the epidermal bar- mens: (1) 0.1% mometasone furoate cream, (2) 0.1% rier undergoes changes, being weakened as a defense triamcinolone acetonide cream, or (3) 0.1% triamci- system. Treatment involves controlling secondary infec- nolone acetonide cream mixed with a topical 20% urea tions, managing itching and excluding any potential cream. Triamcinolone acetonide and 20% urea cream irritating agents. The present work had as objective to were mixed in equal amounts. Each topical agent make a survey of data, in order to better understand (0.25 g per digit referred to finger-tip-unit in humans) the profile of owners of dogs diagnosed with CAD. A was applied twice daily for 14 days. The investigators quantitative questionnaire consisting of 16 questions and dog owners were blinded to the treatment alloca- involving the daily lives of animals and their caregivers tion. Clinical scores were determined by the Canine was applied virtually through Google Docs, in the city Atopic Dermatitis Extent and Severity Index of Rio de Janeiro. A total of 100 responses were (CADESI-4) on experimental days 0, 7, and 14. On obtained. Results show many caregivers still fail to days 7 and 14, mean CADESI-4 score significantly understand the importance of a complete therapeutic reduced in the lesions treated with mometasone (32% approach. Although most prevent ectoparasites (50%), on day 7 and 50% on day 14), triamcinolone (34% on many have the habit of offering varied snacks (51%) to day 7 and 57% on day 14), and triamcinolone plus their dogs, in addition to bathing in a pet shop (50%). urea (29% on day 7 and 54% on day 14) compared The awareness of caregivers of atopic animals regarding with day 0 (p < 0.05). There were no significant inter- optimal care must be made to improve the collabora- group differences in CADESI-4 scores on days 7 and tion that is essential for successful treatment of atopic 14. Thus, triamcinolone acetonide cream, classified as a dogs. medium potency TCS in Japan, may be as effective for Funding Source: Self-funded. AD as the very high potency mometasone furoate. Mix- Conflict of Interest: None declared. ing triamcinolone with 20% urea cream ultimately results in reduction in TCS amount required to achieve an appropriate clinical response. Source of Funding: Self-funded. Conflict of Interest: None declared.

P-119 Bromelain based enzymatic debridement of P-120 deep burns on the paws of a cat Vetch toxicoses in Angus cows fed Vicia sativa L. WALDMAN subsp. sativa (Common Vetch) in New South Dermatology and Allergy Clinic, Haifa, Israel Wales – a case report A bromelain based proteolytic enzyme preparation, A. JORDAN*, B. WATT†, W. BERRY‡ and NexobridTM (MediWound, Yavne, Israel) is used to P. PINCZOWSKI* treat moderate and deep burns in humans. It removes *EMAI, NSW Department of Primary Industries; †LLS, eschar while preserving healthy dermis, thereby, NSW Department of Agriculture; ‡Blayney Vet improving healing and reducing the need for repeated Hospital, NSW, Australia general anesthesia, surgery and autografting. A 13-year- Vetch toxicoses is a well-recognised syndrome of granu- old, male neutered, domestic medium hair cat suffered lomatous eosinophilic dermatitis and systemic disease smoke inhalation, lower urinary tract disease, and sev- associated with ingestion of Hairy Vetch (Vicia villosa), ere burns to all paws in a forest fire. After cleaning the which has also been reported with Narrow-leaf Vetch paws, the bromelain based proteolytic enzyme prepara- (Vicia sativa subsp. nigra). In December 2019, approxi- tion was applied under occlusive dressing for 4 h result- mately 10 cows from a herd of 50 developed cutaneous ing in complete debridement down to healthy dermis. ulcers and crusts over the head, neck, and perineum, To prevent drying, the wounds were treated, without which were erythematous and exudative when lifted. dressings, with sea-buckthorn oil three times daily for There was also some alopecia and scaling. A skin six weeks. The day after the proteolytic enzyme treat- biopsy was submitted, and histopathology showed ment, the cat began to walk cautiously to the litter tray. exudative and ulcerative, perivascular and periadnexal, Pseudoeschar formed within two days. Granulation tis- eosinophilic and histiocytic dermatitis with folliculitis sue appeared after six days and clobetasol propionate and multinucleate giant cells, including Langerhans 0.05% cream was then applied three times daily for six cells. The cattle were being fed Morava hay, variety of weeks. Pain was controlled with buprenorphine Common Vetch (Vicia sativa subsp. sativa), and in light 0.025 mg/kg orally four times daily for the first two of histopathology findings, feeding of Vetch hay was weeks, then meloxicam 0.2 mg/kg orally for one day ceased. Skin lesions largely resolved within two weeks. followed by 0.05 mg/kg for six days. One week after Varieties of Common Vetch have an increasing role in debridement, the cat was alert, walking in the house cropping systems and as livestock fodder in NSW, and and eating well, despite signs of painful paws. Five awareness of a possible role in toxicoses should be weeks later, adhesions in the left hind paw were surgi- raised. cally released. In conclusion, a bromelain based prote- Source of Funding: Local Land Services provided olytic enzyme preparation successfully treated deep paw resources to facilitate the investigation, and AJ is burns in a cat, mitigating the need for intensive wound employed by the Department of Primary Industries, both care. TM within the New South Wales Government Department of Source of Funding: NexoBrid was donated by Medi- Industry, Agriculture and Environment, Australia. Wound. Conflict of Interest: None declared. Conflict of Interest: None declared.

P-121 veterinarian dermatitis VAS on days 0, 7, 14 and 28. Starting from one day after treatment, the percentage A masked, randomized clinical trial evaluating reduction from baseline of Owner-assessed pruritus the efficacy and safety of lokivetmab VAS LS means was significantly higher in the group of compared to saline control in client owned animals treated with lokivetmab versus the control dogs with allergic dermatitis group, with 57.71% reduction compared to 21.78% on Day 28. For all timepoints, the percentage reduction L. VAN BRUSSEL*, H. MOYAERT*, † from baseline of Investigator-assessed skin condition M. ESCALADA*, S. P. MAHABIR and VAS means was significantly higher in the lokivetmab M. R. STEGEMANN* group compared to the control group, with 57.05% *Zoetis Belgium SA, VMRD, Zaventem, Belgium; reduction versus 20.49% on Day 28. Overall, the occur- †Zoetis Inc., VMRD Kalamazoo, MI, USA rence of health events during the in-life phase was com- Interleukin (IL)-31 is an important mediator in atopic parable between both treatment groups. In conclusion, dermatitis and is, as recently reported in humans, also in this study lokivetmab was a safe and efficacious dysregulated in other allergic diseases. To demonstrate treatment for dogs with allergic dermatitis. the efficacy and safety of lokivetmab (an anti-IL-31 Source of Funding: Zoetis Inc, Parsipanny, NJ, USA. monoclonal antibody) for treatment of pruritus associ- Conflict of Interest: All authors are employees of Zoetis ated with allergic dermatitis in dogs.For this study, Inc. dogs (n = 123) were enrolled from 14 practices in Por- Acknowledgements: We thank the clinicians who enrolled tugal, Hungary, France and Germany. Animals were at dogs in this study and performed the clinical investiga- least moderately pruritic at enrolment and a presump- tions: Jean-Michel Arquilliere, Emmanuel Bensignor, tive diagnosis of allergic dermatitis was made. Animals Laurent Bourdenx, Vincent Mahe, Isabelle Papadopoulo, were randomized to placebo (saline, n = 62) or lokivet- Annette Feische, Elisabeth Koldt, Petra Wittmann, mab (1–3.3 mg/kg, n = 61). Dogs were treated once on Balazs Capari, Laszl o Kiraly, Szilard Kraxner, Krisztina day 0. Pruritus was assessed daily by the owner until Palfi, Joao Araujo, Catarina Duarte, Adriana Estrela day 7 and weekly thereafter through a Visual Analogue and Manuela Mangas. Scale (VAS) by Hill. Skin condition was assessed by a