第32届欧洲兽医皮肤科大会(在线)2021年9月16-16日摘要-（1）

王帆

32nd European Veterinary Dermatology Congress (online) 16–16 September 2021 Abstracts

第32届欧洲兽医皮肤科大会(在线)2021年9月16-16日摘要

翻译：王帆 校对：刘欣

Are images worth a thousand words? Testing a video for owners’ education in canine atopic dermatitis

图片胜过千言万语吗?测试一段用作宠主教育的犬特应性皮炎视频

FERNANDES*, A. CAVACO† , H. PEREIRA*, M. PINTO*, A. BIZARRO* and A. LOURENCO* *CIISA, Faculty of Veterinary Medicine, † IMed. ULisboa, Pharmacy Faculty, University of Lisbon, Lisboa, Portugal

Successful management of canine atopic dermatitis (cAD) requires effective pet owner education considering its challenging and complex nature. Given the lack of proven effective owner education strategies, we wanted to create an effective and appealing cAD owner education tool of feasible application in the clinical context. An eight week prospective randomised controlled study was conducted. Our study hypothesis was that the video-based educational intervention would be noninferior or equivalent to the control group (standard verbal educational intervention). Participants were randomly assigned (1:1) to either the control (CG) or the intervention group (IG). In both groups, cAD knowledge scores and clinical outcomes were measured at the beginning and end of the study. Additionally, owners’ adherence and perceived utility and appeal ratings were measured at the end of the study. The educational intervention’s outcome was defined by a cAD knowledge score improvement, provided that the end score was superior to the positive scale’s central value. A significant association (P < 0.01) was found between educational intervention outcome and group, with a higher percentage of success in the IG [CG 6/13 (46.15%); IG 15/16 (93.75%)]. The differences found in the clinical outcomes, utility and appeal ratings, and owners’ adherence were not statistically significant. This novel study establishes a positive impact of videobased education on therapeutic owner education regarding cAD, confirming its great potential as an effective way to deliver pet owner’s education. It also provides general veterinarians with the opportunity to achieve good-quality cAD owner education in a time-efficient way.

考虑CAD的困难和复杂性。要想成功的管理犬特应性皮炎(cAD)需要有效的宠主教育。由于缺乏行之有效的宠主教育策略，我们希望创建一个有效的、有吸引力的cAD宠主教育工具，可在临床环境中应用。进行了一项为期8周的前瞻性随机对照研究。我们的研究假设是基于视频的教育干预将不低于或等同于对照组(标准语言教育干预)。参与者被随机(1:1)分配到对照组(CG)或干预组(IG)。在两组中，在研究开始和结束时测量cAD知识评分和临床结果。此外，在研究结束时，还测量了宠主的忠诚度、感知效用和吸引力评级。教育干预的结果是通过cAD知识得分的提高来定义的，前提是最终得分优于正量表的中心值。显著关联(P＜0.01)， IG组的成功率较高[CG 6/13 (46.15%);IG 15/16(93.75%)。在临床结果、效用和吸引力评级以及宠主的依从性方面发现的差异没有统计学意义。这项新研究建立了基于视频的教育对治疗性CAD的主人教育的积极影响，证实了它作为一种有效的方式来提供宠物主人教育的巨大潜力。它还为普通兽医提供机会，以时间有效的方式实现高质量的cAD宠主教育。

Source of funding: CEVA Saude Animal

Conflicts of interest: None declared.

Transcriptome signatures in peripheral blood and skin cells of dogs with atopic dermatitis

特应性皮炎犬外周血和皮肤细胞的转录组特征

ALTUNBULAKLI*, N. FISCHER† , C. AKDIS‡ , C. FAVROT† and A. ROSTAHER† *Lund University, Department of Immunotechnology, Lund, Sweden † Clinic for Small Animal Internal Medicine, Vetsuisse Faculty Zurich, Zurich, Switzerland ‡ Swiss Institute of Allergy and Asthma Research (SIAF), Davos, Switzerland

Canine atopic dermatitis (AD) is a multifactorial disease characterised by skin barrier and immune cell dysfunction. We aimed to study the state of circulating immune cells in atopic dogs, a relatively understudied area of AD. RNA was isolated from peripheral blood mononuclear cells (PBMCs) and nonlesional skin tissue collected from seven client-owned atopic dogs with active disease and 11 healthy dogs. RNA-sequencing was performed using the Illumina Hi-Seq 2500 platform. Gene Ontology (GO) term enrichment and pathway analysis were performed with the GOSEQ R package and Enricher platform, respectively. The comparison of PBMC RNA-seq data from atopic and healthy dogs revealed a distinct transcriptome expression in AD, with 1,604 significantly regulated genes (P < 0.01). The GO analysis indicated an increased expression of genes belonging to the regulation of mitogen-activated protein kinase pathways, immunoglobulin production and general inflammatory response. Among the most differentially expressed genes were CD80, CD83, ICAM1, PD-L1 and IRF1, suggesting a breaking of inflammatory homeostasis in the peripheral blood. When comparing skin tissue RNA-seq between atopic and healthy dogs, we identified 804 significantly regulated genes (P < 0.01). Upregulated genes in AD skin include antimicrobial peptides S100A8 and S100A9, genes belonging to production of matrix metalloproteinases (TIMP3, MMP1), as well as enzymes and transcription factors regulating tissue inflammatory response. We herein show the dysregulation of immune cells in the peripheral blood and in the skin inflammation in AD. A closer analysis of RNA-seq is a step towards unravelling the complete disease pathogenesis of AD and identifying possible biomarkers.

犬特应性皮炎(AD)是一种多因子疾病，以皮肤屏障和免疫细胞功能障碍为特征。我们的目的是研究特应性疾皮炎犬体内循环免疫细胞的状态，AD是一个相对缺乏研究的领域。从7只家养的活跃性特应性皮炎患犬和11只健康犬的外周血单个核细胞(pmcs)和非病变性皮肤组织中分离出RNA。RNA测序采用Illumina Hi-Seq 2500平台。分别使用GOSEQ R包和Enricher平台进行基因本体论(GO)术语富集和通路分析。比较AD犬和健康犬的PBMC RNA-seq数据，发现AD中有不同的转录组表达，有1604个显著调节的基因(P <0.01)。氧化石墨烯分析表明，属于丝裂原活化蛋白激酶通路调控、免疫球蛋白产生和一般炎症反应的基因表达增加。表达差异最大的基因有CD80、CD83、ICAM1、PD-L1和IRF1，提示外周血炎症稳态被打破。当比较AD和健康犬的皮肤组织RNA-seq时，我们发现804个显著调节的基因(P <0.01)。AD皮肤中上调的基因包括抗菌肽S100A8和S100A9，属于基质金属蛋白酶(TIMP3, MMP1)的产生基因，以及调节组织炎症反应的酶和转录因子。我们在此显示了AD患犬外周血和皮肤炎症中免疫细胞的失调。对RNA-seq的进一步分析是阐明AD完整的疾病发病机制和识别可能的生物标志物的一步。

Source of funding: Swiss National Fund. Conflicts of interest: None declared.

Bedlington terriers diagnosed with familial footpad hyperkeratosis are carriers of an FAM83G mis-sense variant

被诊断为家族性爪垫过度角化症的贝灵顿㹴是一种FAM83G错义变种的携带者

MAKRI*, K. VARJONEN† , C. WALKER‡ , D. OZDEMIR§ , J. SCHOENEBECK§ and D. GOW* *R(D)SVS and The Roslin Institute, The University of Edinburgh, Edinburgh, UK † Anicura, Stockholm, Sweden ‡ TheSkinVet, Essex, United Kingdom

Hereditary footpad hyperkeratosis (HFH) is a genetically heterogenous group of diseases described in various breeds, notably Irish terriers and the Kromfohrlander. Genetic mutations in these breeds are published and prebreeding screening tests available. Affected dogs develop hyperkeratotic pads that become deeply fissured and secondarily infected, thus leading to significant pain and discomfort. Because HFH is a hereditary disease, treatment is symptomatic (e.g. the use of softening agents). Bedlington terriers also have been noted to develop HFH. Lesions in these dogs have been seen at an early age,indicating a genetic predisposition. The aim of this study was to assess the genetic profile of Bedlington terriers with FH to identify an underlying genetic defect responsible for the hyperkeratosis. Three Bedlington terriers were clinically diagnosed with FH. Saliva swabs were collected from each dog and genomic DNA extracted according to the manufacturer’s protocol. Each sample was genotyped for the FAM83G:c.155G>C variant by Sanger sequencing. All three affected dogs were homozygous for the FAM83G risk allele, while two control Bedlington terriers were negative for this mutation. All affected dogs were diagnosed with HFH between the ages of six and nine months, with the first signs of disease noted by the owner as paw-licking and lameness. Affected dogs responded to the use of regular hydrating agents applied to the paws. The finding of a FAM83G mutation as a cause for HFH in the Bedlington terrier may allow for genetic screening to be offered in this breed to subsequently reduce the frequency of occurrence.

遗传性爪垫过度角化症(HFH)是一种遗传异质性的疾病在各种品种都有描述，特别是爱尔兰㹴和克龙弗兰犬。这些品种的基因突变已公布，育种前筛选试验可用。患犬发展爪垫过度角化，出现深层开裂和继发感染，从而导致显著的疼痛和不适。由于HFH是一种遗传性疾病，治疗是对症的(例如使用软化剂)。贝灵顿㹴也被发现有HFH。这些犬的病变在早期就已经看到，表明遗传易感性。本研究的目的是评估贝灵顿㹴FH的遗传谱，以确定导致过度角化的潜在遗传缺陷。三只贝德灵顿犬被临床诊断为FH。收集每只犬的唾液拭子，并根据制造商的协议提取基因组DNA。通过Sanger测序对每个样本进行FAM83G: C . 155g>C型基因分型。所有3只患犬都是FAM83G风险等位基因的纯合子，而2只对照贝灵顿㹴对这种突变是阴性的。所有患犬在6到9个月之间被诊断出患有HFH，主人注意到的第一个症状是舔爪和跛行。患犬爪部使用常规保湿剂有效。在贝灵顿㹴中发现FAM83G突变是HFH的一个原因，这可能允许对该品种进行基因筛查，从而减少发生的频率。

Source of funding: Fiona and Ian Russell Clinical Seed Corn grant, R(D)SVS. Conflicts of interest: None declared.

Compliance and adverse effects of oral ciclosporin A administration in cats: a four-year retrospective survey of long-term administration in referral practice

口服环孢素A给药猫的依从性和不良反应:一项关于转诊中心中长期给药的四年回顾性调查

DELEPORTE*, A. BRIAND† and P. PRELAUD† *AniCura LorraineVet, Ludres, France † ADVETIA CHV, Velizy Villacoublay, France

The aim of the study was to assess the compliance of ciclosporin treatment in cats. Thus, we performed a survey of 42 owners who had used ciclosporin for their cats. The cat population was composed of 30 domestic short hair (DSH), five Maine coon, two Birman and five other breeds (20 males, 22 females; median age 5.5 years). Ciclosporin oral suspension was administered directly in 75% and with food in 25% of cases. The administration was considered easy, difficult and impossible in 43%, 55% and 2% of cases, respectively. The treatment was disrupted in 50% of cases for the following reasons: difficulty of administration (9/42), failure (4/42), price (2/42) and other causes (2 deaths, 2 tumours, 1 secondary adverse effect and one lack of compliance). Adverse events were reported in 59% of cases as follows: vomiting-diarrhoea (8/42), ptyalism (8/42), dysorexia-anorexia (6/42), gingival hyperplasia (1/42), and combined vomiting, diarrhoea and ptyalism (2/42). Behavioural problems were reported in 64% of cases as follows: hiding (7/42), scared of owner (10/42), modification of sleeping or playing activity (6/42), urine/faeces out of the litter (2/42), aggression towards the owner (2/42) and all of the above (1/42). In total, 57% presented both physical adverse events and behavioural problems, and only 7% presented either physical adverse events or behavioural problems. Interestingly this study suggests that behavioural problems were underestimated in the cats, either because of therapeutic failure or lack of compliance. Larger scale studies are needed to confirm these preliminary results.

本研究的目的是评估环孢素治疗对猫的依从性。因此，我们对42名曾为猫使用环孢素的猫主进行了调查。猫群由30只家养短毛猫(DSH)、5只缅因猫、2只伯曼猫和5只其他品种的猫组成(雄性20只，雌性22只;平均年龄5.5岁)。75%的病患直接服用环孢素口服混悬液，25%的病患与食物一起服用。分别有43%、55%和2%的病例认为给药容易、困难和不可能。50%的病例因以下原因中断治疗:给药困难(9/42)、失败(4/42)、价格(2/42)和其他原因(2例死亡、2例肿瘤、1例继发性不良反应和1例缺乏依从性)。59%的病例报告的不良事件为:呕吐-腹泻(8/42)、流涎(8/42)、食欲不良-厌食症(6/42)、牙龈增生(1/42)和合并呕吐、腹泻和流涎(2/42)。64%的病例报告的行为问题如下:躲藏(7/42)，害怕主人(10/42)，改变睡觉或玩耍活动(6/42)，在窝里小便(2/42)，对主人的攻击(2/42)和以上所有的(1/42)。总的来说，57%同时出现了身体不良反应和行为问题，只有7%出现了身体不良反应或行为问题。有趣的是，这项研究表明，由于治疗失败或缺乏依从性，猫的行为问题被低估了。需要更大规模的研究来证实这些初步结果。

Source of funding: Self-funded. Conflicts of interest: None declared.

A prospective, controlled, multicentric and doubleblinded study of the efficacy of combined treatment of allergen-specific immunotherapy and lokivetmab for the treatment of canine atopic dermatitis: a pilot study

一项前瞻性、控制性、多中心和双盲研究:过敏原特异性免疫疗法和洛基维特单抗联合治疗犬特应性皮炎的疗效:一项初步研究

L. RAMIO-LLUCH  * and A. PUIGDEMONT† *LETI Pharma, Barcelona, Spain † Universitat Autonoma de Barcelona, Barcelona, Spain

The objective of this pilot, prospective, double-blinded, controlled, multicentric study was to evaluate the efficacy of allergen-specific immunotherapy (ASIT) given concomitantly with lokivetmab in dogs with atopic dermatitis (AD) during one year. Sixteen privately owned, mite-allergic dogs were enrolled in this study. Dogs were treated with subcutaneous ASIT (treatment group; n = 7) or placebo (control group, n = 9) for 12 months concomitantly with lokivetmab before, and after one and two months of starting ASIT. Canine Atopic Dermatitis Extent and Severity Index (CADESI) and pruritus Visual Analog Scale (pVAS) scores were obtained at month (m)0, m2, m5, m8 and m12. Lokivetmab was the rescue medication after the two months period. Significant differences between groups were observed in the CADESI values (P = 0.019). In the lokivetmab-treated group, values significantly decreased from 33.4  13.2 to 2.9  2.6 (mean  SD) with a mean improvement of 91%. In the control groups, they decreased from 32.7  8.3 to 12.9  10.4 (58% improvement). The pVAS values also decreased during the treatment in both groups, ranging from 6.7  2.0 to 2.6  2.5 in the control group (47% improvement) and from 7.1  1.5 to 1.3  1.5 in the treated group (82% improvement). Significant differences were not observed. This pilot study revealed that combined treatment with lokivetmab at the beginning of ASIT leads to a faster improvement of AD and improves owner compliance. Further evaluation of a larger population of dogs treated with the combined regimen should be conducted to verify these observations.

该试验是一项前瞻性、双盲、对照、多中心研究，目的是评估过敏源特异性免疫疗法(ASIT)联合洛基维特单抗治疗特应性皮炎(AD)犬一年的疗效。这项研究招募了16只家养的螨虫过敏犬。采用皮下ASIT治疗(治疗组;n = 7)或安慰剂(对照组，n = 9)，在ASIT开始前和开始1个月和2个月后，联合洛基维特单抗治疗12个月。犬特应性皮炎严重程度指数(CADESI)和瘙痒视觉模拟量表(pVAS)评分分别于(m)0、m2、m5、m8和m12月进行。洛基维特单抗是两个月后的使用药物。CADESI值组间差异有统计学意义(P = 0.019)。在洛基维特单抗治疗组，数值显著下降，从33.4 13.2下降到2.9 2.6(平均SD)，平均改善91%。在对照组中，他们从32.7 8.3下降到12.9 10.4(58%改善)。两组治疗期间pVAS值也有所下降，对照组从6.7 2.0到2.6 2.5(改善47%)，治疗组从7.1 1.5到1.3 1.5(改善82%)。未观察到显著差异。这项初步研究显示，在ASIT开始时联合使用洛基维特单抗可更快地改善AD，并提高病患的依从性。为了验证这些观察结果，应该对更大数量的犬进行进一步的评估。

Source of funding: LETI Pharma, Barcelona, Spain. Conflicts of interest: LR is an employee of LETI Pharma; AP has received unrelated funding from LETI Pharma.

Characterization of clinical phenotype (cutaneous lesion score and central sensitization) and comparison of serological allergen-specific immunoglobulin E and intradermal testing in a colony of atopic-like cats

临床表型的特征(皮肤病变评分和中枢致敏)，以及在一群特应性皮炎猫科动物中进行的血清学过敏原特异性免疫球蛋白E和皮内试验的比较

J. LEFRANCOIS*, E. TRONCY† , A. GONZALES‡ , M. VISSER‡ , C. OTIS† , M. MOREAU§ , J.P. PELLETIER§ , J. MARTEL-PELLETIER§ and F. SAUVE* *Faculte de m  edecine v  et  erinaire, Universit  e de Mon-  treal, St-Hyacinthe, Canada  † Groupe de recherche en pharmacologie animale du Quebec (GREPAQ), St-Hyacinthe, Canada  ‡ Zoetis, Kamalazoo, MI, USA § Osteoarthritis Research Unit, University of Montreal Hospital Research Centre, Montreal, Canada

Intradermal testing (IDT) and allergy serology testing (AST) are used in patients with feline atopic syndrome (FAS), and the relationship between these two tests is unknown. Central neural sensitization has been demonstrated in humans with chronic pruritus and cats with chronic osteoarthritis pain. We evaluated the relationship between IDT and AST results in a colony of cats with FAS, with or without skin lesions, and characterised their somatosensory profile. Twenty-four cats were divided into three groups: (i) FAS with skin lesions (16); (ii) FAS without skin lesions (4); and (iii) control (4). With group blinding the following were evaluated: a complete physical examination, Scoring Feline Allergic Dermatitis (SCORFAD) calculation, IDT (Stallergenes Greer, London, ON, Canada), AST (Heska, Loveland, CO, USA), somatosensory profile determined by punctate tactile withdrawal threshold test (using von Frey aesthesiometer on paw plantar surface) and response to mechanical temporal summation (RMTS). All statistical analyses were performed with an alpha value set at 0.05. For the AST results, the allergens were grouped into seven categories. In the first group, the median SCORFAD was 4. Results showed a significant relationship between the IDT and AST only for the ‘mites’ category [v2 1 = 10.9, P = 0.003]. The RMTS was significantly (P < 0.049) lower in both FAS groups [10.9(3.6), 8.9(1.4)] compared to control [13.6(2.3)]. These results reinforce the lack of relationship between IDT and AST. This is the first report investigating a possible role of central sensitization in FAS, which could help better understand the pruritus pathways and open new horizons for treatment.

皮内试验(IDT)和过敏血清学试验(AST)用于特应性综合征(FAS)患猫，但这两种试验之间的关系尚不清楚。患有慢性瘙痒症的人和慢性骨关节炎疼痛患猫已被证实具有中枢神经敏感化。我们在一群FAS患猫中评估了IDT和AST结果之间的关系，并对它们的躯体感觉特征进行了描述。将24只猫分为3组:(i) FAS伴皮肤病变(16);(ii)无皮肤病变的FAS (4);(3)对照组(4)。采用盲法对以下情况进行评价:全面体检，评分猫过敏性皮炎(SCORFAD)计算，IDT，通过点状触觉退缩阈值测试(使用足跖面von Frey触觉计)和机械时间总和(RMTS)反应来确定躯体感觉套组。将alpha值设置为0.05，进行所有的统计分析。对于AST结果，过敏原被分为7类。在第一组中，平均SCORFAD为4。结果仅在螨类中IDT与AST存在显著相关性[v2 1 = 10.9, P = 0.003]。RMTS显著(P <0.049)的FAS组[10.9(3.6)，8.9(1.4)]低于对照组[13.6(2.3)]。这是首个研究中枢致敏在FAS中可能作用的报告，这将有助于更好地理解瘙痒通路，并为治疗开辟新的视野。

Source of funding: Zoetis Inc., Kalamazoo, MI, USA. Conflicts of interest: AG and MV are employees of Zoetis Inc.

Single myringotomy for the treatment of otitis media in cats: a retrospective study

单一鼓膜切开术治疗猫中耳炎:一项回顾性研究

S. DELEPORTE* and P. PRELAUD  † *LORRAINEVET, Ludres, France † CHV ADVETIA, Velizy-Villacoublay, France

Otitis media (OM) is common in cats, yet, except for polyp-associated OM, data on medical treatment are lacking. We retrospectively studied 24 cats with magnetic resonance imaging (MRI)- or computed tomography (CT)- confirmed OM without polyps or neoplasia. Most cats (58%) presented with a bilateral OM. Both parts of the bulla were involved in 97% of cases. Clinical signs included head tilt in 11/24 cats (54%), Horner syndrome in 7/24 (29%), ataxia or otalgia in 2/24 (9%), otitis externa in 5/24 (21), and nystagmus and facial paralysis in 1/24 (4%). Cytological examination found middle ear effusion dominated by neutrophils. Cocci were identified on cytological evaluation in 20/38 samples (53%) and rods in 2/38 samples (5%). The bacterial culture was positive in half of the samples, with Pseudomonas sp., Pasteurella multocida, Staphylococcus felis, S. schleiferi, S. pseudintermedius and Serratia marcescens isolated. All cats were treated with a single myringotomy, soft bulla flushing (0.5–2 mL saline), oral prednisolone (1 mg/kg/d for seven to 10 days) and one month of systemic antibiotics chosen according to the susceptibility testing. Seventeen cats (71%) were clinically cured 60–240 days after starting treatment, while one cat was healthy except for a mild head tilt. One cat was euthanised because of failure to control a Pseudomonas infection; another died because of complications of anaesthesia. The four remaining cats were cured after ventral bulla osteotomy. This simple technique with only one myringotomy and mild ear flushing offers a very practical, safe and efficient method to treat suppurative OM in cats.

中耳炎(OM)在猫身上很常见，但除与息肉相关的OM外，缺乏关于医学治疗的数据。我们回顾性研究了24只经磁共振成像(MRI)或计算机断层扫描(CT)证实无息肉或肿瘤的OM猫。大多数猫(58%)表现为双侧OM。97%的病例都涉及鼓泡的两个部分。临床症状包括头倾斜11/24(54%)，霍纳综合征7/24(29%)，共济失调或耳痛2/24(9%)，外耳炎5/24(21)，眼球震颤和面瘫1/24(4%)。细胞学检查发现中耳积液以中性粒细胞为主。在20/38份样本(53%)中细胞学鉴定出球菌，在2/38份样本(5%)中鉴定出杆菌。半数样品细菌培养阳性，分离出假单胞菌、多杀性巴氏杆菌、猫葡萄球菌、施莱费氏葡萄球菌、假中间型葡萄球菌和粘质沙雷氏菌。所有猫均予鼓膜切开、轻柔鼓泡冲洗(0.5-2 mL生理盐水)、口服泼尼松龙(1 mg/kg/d，持续7-10天)和根据药敏试验选择的全身抗生素治疗1个月。17只猫(71%)在开始治疗60 - 240天后临床治愈，其中1只猫除轻度头倾斜外，其余均为健康。一只猫因为无法控制假单胞菌感染而被安乐死;另一只死于麻醉并发症。其余4只猫经腹侧鼓泡截骨术后痊愈。这种简单的技术只需要一次鼓膜切开术和轻微的耳冲洗，为治疗猫的化脓性OM提供了一种非常实用、安全、有效的方法。

Source of funding: Self-funded. Conflicts of interest: None declared.

In vitro antibacterial effect and cytotoxicity on canine keratinocytes of PurO3 Ozonated Olive Oil

PurO3臭氧化橄榄油对犬角质细胞的体外抑菌作用及细胞毒性研究

N. MAHEAS, B. CHATAIGNER, N. MILHAU, D. PIN and M. MOSCA VetAgro Sup, Marcy l’etoile, France  

In the current antibacterial resistance context, alternative therapies to decrease the use of antibiotics are necessary. PurO3 Ozonated Olive Oil (O3-Oil) is a skin ointment with antiseptic, healing and anti-inflammatory properties that could be useful in treating canine pyoderma. This in vitro study aims to prove the antibacterial effect of O3- Oil on Staphylococcus pseudintermedius and its lack of toxicity on canine keratinocytes. The antibacterial effect of O3-Oil at different concentrations (2.5%, 5%, 7.5% and 10% incorporated into TSA agar plates) was tested on nine S. pseudintermedius strains obtained from dogs with pyoderma (three sensitive to all antibiotics tested, three resistant to one or two antibiotics, three multiresistant). Canine progenitor epidermal keratinocytes (CPEK) were incubated with O3-Oil at different concentrations (0%, 25%, 50%, 75% and 100% diluted with virgin olive oil) for 2, 4 and 6 h, and cell viabilities were determined by a colorimetric CCK8 assay to evaluate toxicity. Results showed a minimal inhibitory concentration varying between 5% and 10% depending on the strain tested. All bacterial strains of S. pseudintermedius were completely inhibited at 10% O3-Oil within the culture medium. The toxicity assay showed no significant difference of cell viability between virgin olive oil and the different concentrations of O3-Oil tested, even pure (P > 0.05 at each time and each concentration, Student’s t-test). O3-Oil is noncytotoxic to keratinocytes and has a strong in vitro antibacterial effect. Further research is needed to demonstrate in vivo efficacy of the product for topical treatment of canine pyoderma.

在目前的抗菌药物耐药性背景下，替代疗法减少抗生素的使用是必要的。PurO3臭氧化橄榄油(O3-Oil)是一种皮肤软膏，具有杀菌、愈合和消炎的特性，可以用于治疗犬脓皮病。本体外实验旨在证明O3-油对假中间葡萄球菌的抑菌作用及对犬角质细胞无毒性作用。来自脓皮病患犬身上的9株假中间型葡萄球菌(3株对所有抗生素敏感，3株对1-2种抗生素耐药，3株多重耐药)进行了不同浓度的o3-油(TSA琼脂板中加入2.5%、5%、7.5%和10%)的抑菌试验。将犬表皮角质细胞(CPEK)与不同浓度的o3-油(0%、25%、50%、75%和100%用初榨橄榄油稀释)孵育2、4和6 h，采用CCK8比色法测定细胞活力，以评价其毒性。结果显示最小抑制浓度变化在5%和10%之间取决于菌株测试。在10%的o3-油浓度下，所有菌株均被完全抑制。毒性试验表明，初榨橄榄油和不同浓度的O3-Oil，即使是纯的(P >0.05，st检验)无显著差异。o3-油对角质细胞无细胞毒性，具有很强的体外抗菌作用。还需要进一步的研究来证明该产品在外部治疗犬脓皮病的体内疗效。

Source of funding: Self-funded. Conflicts of interest: None declared.

Effectiveness of fluralaner against ticks infesting horses (Equus caballus)

氟雷拉纳对寄生在马身上的蜱虫的效果

G. SHEINBERG*, C. ROMERO*, A. CORDERO† , L. BAUTISTA‡ and R. HEREDIA§ *Dermatologia Especializada CVM, Mexico City, Mexico † VETDERM, Guadalajara, Mexico ‡ Centro Universitario UAEM, AMECAMECA EDOMEX, Mexico § DERMAVET, EDOMEX, Mexico

Tick control in horses usually is achieved through the application of different acaricides and there is no report of using isoxazolines. The study objective was to evaluate the efficacy of fluralaner (Bravecto, MSD; Kenilworth, NJ, USA) against ticks in horses. Five females and five males were used in the trial, with an average age of 12 ( 5) years, weighing 432 ( 12) kg. Selected animals were evaluated for the presence of ticks and did not receive any acaricides 30 days before the trial. All horses were treated with fluralaner 25 mg/kg. Ticks were counted and identified morphologically on day (D)1, D7, D14, D30, D45, D60 and D90. No other treatment was administered during the study. Data were analysed using the Wilcoxon matched pairs test with an alpha of 0.05 for comparison between post-treatment measurements. Two tick species were identified: Amblyomma cajennense and Rhipicephalus microplus. The most significant number of ticks were found in the ears (55.63%), followed by the head (23.23%) and the rest of the body (21.12%). On D7, tick numbers were significantly reduced by 78.67% (P = 0.0001). On D14, there was a continued decrease in tick numbers compared to initial counts; tick numbers were reduced by 94% (P = 0.0001). By D30, tick numbers were reduced by 99.33% (P = 0.006). From D30 to D90, no ticks were found showing the sustained activity of fluralaner over time. No adverse effects were observed during clinical trial. Based on these data, fluralaner appears to provide effective control of A. cajennense and R. microplus ticks for ≥90 days.

对马的蜱虫控制通常是通过使用不同的杀螨剂来实现的，目前还没有使用异恶唑啉的报告。研究目的是评价氟雷拉纳(Bravecto, MSD;Kenilworth, NJ，美国)对马身上的蜱虫作用。试验对象为5名雌性和5名雄性，平均年龄为12岁( 5岁)，体重为432 ( 12)公斤。被选中的动物在试验前30天没有接受任何杀螨剂，并被评估是否存在蜱虫。所有马均给予25 mg/kg的氟雷拉纳治疗。第1天、第7天、第14天、第30天、第45天、第60天和第90天对蜱虫进行形态学计数和鉴定。在研究期间没有进行其他治疗。数据采用Wilcoxon配对检验，alpha值为0.05，用于比较治疗后测量值。鉴定出两种蜱种:钝头蜱(Amblyomma cajenense)和微头蜱(头蜱)。耳中蜱的数量最多(55.63%)，其次为头部(23.23%)和身体其他部位(21.12%)。第7天，蜱数显著减少78.67% (P = 0.0001)。在第14天，蜱虫数量较开始时持续减少;蜱虫数量减少94% (P = 0.0001)。D30时，蜱虫数量减少了99.33% (P = 0.006)。在D30 ~ D90期间，未发现有持续活动的蜱虫。临床试验期间未观察到不良反应。根据这些数据，氟雷拉纳控蜱可有效控制卡延花蜱和微小牛蜱≥90 d。

Source of funding: Self-funded. Conflicts of interest: None declared.

The comparative cerumenolytic activity of otic preparations, an in vitro study

耳部制剂耵聍溶解活性的比较，体外研究

N. MILANESI*, G. GHIBAUDO* and T. DELLA MURA† *Clinica Veterinaria Malpensa, Samarate (Varese), Italy † ICF S.r.l., Palazzo Pignano, Italy

The canine ear canal has an abundant presence of sebaceous and ceruminous glands. Ceruminous otitis is a very common pathology in dogs. The cerumenolytic activity of some otic preparations is valuable in the management of this condition. The study aimed to evaluate the in vitro wax removal efficacy of various veterinary otic preparations used in Europe. To evaluate the effectiveness of the otic preparations (Otoprof, Otoact, Epiotic, Diclorex oto, Otifree, Aurinet, Malacetic, Otodog and Otolane) an analytical and experienced protocol was used. Standardised synthetic cerumen (SCC) was prepared, weighed in test tubes and left to solidify. An aliquot of the otological product equal to 2 mL was added. The tubes were shaken moderately at a temperature of 35°C for 5 min and subsequently inverted. After 24 h each test tube was weighed, and another 2 mL product were added. Five washes per tube were repeated and the test was performed in triplicate. A total of 99.4% of removed cerumen was obtained only with the product containing carbamide peroxide and dioctyl sodium sulfosuccinate (Otoprof, ICF S.r.l.; Palazzo Pignano, Italy). Good cerumenolytic activity (65%) was obtained from the product containing squalene (Otoact, ICF S.r.l.). Other products containing disodium EDTA, PCMX, diethylhexyl sodium sulfosuccinate such as Epiotic (Virbac S.r.l.; Milano, Italy) showed less (7%) cerumenolytic activity and the remaining ones just 5% to 1% activity.

犬耳道有丰富的皮脂腺和耵聍腺。耵聍性耳炎是犬的一种常见病。一些耳部制剂的耵聍溶解活性对这种情况的治疗中是有价值的。本研究旨在评价欧洲使用的各种兽医耳制剂的体外除耵聍效果。为了评价耳部科制剂(Otoprof, Otoact, Epiotic, Diclorex oto, Otifree, Aurinet, Malacetic, Otodog and Otolane)的有效性，采用分析和经验的方案。标准化的合成耳垢(SCC)被制备出来，在试管中称量，然后让它固化。加入等于2毫升的耳科产品。在35℃温度下适度摇晃试管5分钟，然后倒置。24 h后称量每个试管，再加入2 mL产品。每管重复5次洗涤，试验进行3次。仅用含有过氧化脲和二辛基磺基琥珀酸钠(Otoprof, ICF S.r.l.; Palazzo Pignano、意大利)。含角鲨烯(Otoact, ICF S.r.l.)的产品具有良好的溶垢活性(65%)。其他含有EDTA二钠，PCMX，二乙基己基磺基琥珀酸钠的产品，如耳漂 (维克 S.r.l.;意大利，米兰)表现出更低的(7%)溶垢活性，其余的活性仅为5%至1%。

Source of funding: ICF S.r.l. Conflicts of interest: NM is an ICF S.r.l. employee, TDM is ICF S.r.l. Scientific Manager and GG have received consultancy and speaker fees from ICF S.r.l.

Stability of N-acetylcysteine (NAC) with Tris-EDTA solution and in combination with dexamethasone sodium phosphate in aqueous solution for 50 days

n -乙酰半胱氨酸(NAC)与Tris-EDTA溶液及与地塞米松磷酸钠溶液联合作用50天的稳定性

N. MILANESI*, G. GHIBAUDO* and T. DELLA MURA† *Clinica Veterinaria Malpensa, Samarate (Varese), Italy † ICF S.r.l., Palazzo Pignano, Italy

The objective of this study was to evaluate the stability of a formulation containing N-acetylcysteine (NAC) + TrisEDTA (Tris-NAC, ICF S.r.l.; Palazzo Pignano, Italy) and the compatibility between the dexamethasone sodium phosphate in aqueous solution, 2 mg/mL injectable formulation (Dexadreson, MSD Italia S.r.l.; Roma, Italy) and TrisNAC for 50 days. To evaluate the compatibility between dexamethasone and Tris-NAC, redox titrations were performed to evaluate the NAC content in the formulations. Two tests were performed: (i) stability assessment of Tris-NAC mixing 100 mL formulation with 5 mL dexamethasone sodium phosphate solution (2 mg/mL); and (ii) stability assessment of the Tris-NAC formulation without any additions. The two formulations were stored for 50 days at 25°C and 5°C and titrations were performed every 10 days to evaluate the NAC content in the formulation. The experimental data demonstrated the NAC titre in the two formulations to be comparable. In conclusion, after 50 days at 25°C and 5°C, both formulations maintained almost unchanged NAC titration compared to the start point.

本研究的目的是评价含n-乙酰半胱氨酸(NAC)+TrisEDTA的制剂的稳定性(Tris-NAC, ICF S.r.l;地塞米松磷酸钠在2 mg/mL注射剂(Dexadreson, MSD Italia S.r.l.;罗马，意大利)和TrisNAC为期50天。为评价地塞米松与Tris-NAC的相容性，采用氧化还原滴定法评价制剂中NAC的含量。进行了两项试验:(i)稳定性评估Tris-NAC混合100 mL配方与5 mL地塞米松磷酸钠溶液(2 mg/mL);(ii)不添加任何添加剂的Tris-NAC配方的稳定性评估。两种制剂分别在25℃和5℃保存50天，每10天进行一次滴定，测定制剂中NAC含量。实验数据表明，两种剂型的NAC滴度具有可比性。综上所述，在25℃和5℃条件下，50 d后，两种剂型的NAC滴定量与起始点基本保持不变。

Source of funding: ICF S.r.l. Conflicts of interest: NM is an ICF S.r.l. employee, TDM is ICF S.r.l. Scientific Manager, and GG received consultancy and speaker fees from ICF S.r.l.

Tris-NAC (Tris-EDTA + N-acetylcysteine) activity against biofilm production, an in vitro study

Tris-NAC (Tris-EDTA+N-乙酰半胱氨酸)对生物膜产生的体外活性研究

N. MILANESI‡ , G. GHIBAUDO† AND T. DELLA MURA‡ *Clinica Veterinaria Malpensa, Samarate (Varese), Italy † ICF S.r.l., Palazzo Pignano, Italy

The aim of the study was to assess the efficacy of product containing N-acetylcysteine with Tris-EDTA (TrisNAC, ICF S.r.l.; Palazzo Pignano, Italy) in preventing the formation of biofilm and aiding its disintegration. The study of biofilm growth inhibition by this formulation, conducted by the Clever Bioscience S.r.l. laboratory (Pavia, Italy), was evaluated using the microplate biofilm formation test against three micro-organism strains: Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923) and Malassezia pachydermatis (DSM 6172). The same three micro-organisms were grown in microtitre plates and the biofilm method evaluated by Microtiter Dish Biofilm Formation Assay. Biofilm growth and production were assessed at two different time points (8 h, 24 h) for bacteria and at three different times for yeasts (8 h, 24 h, 48 h). Subsequently, the Tris-NAC formulation, at different dilutions, was added to the same plates to evaluate the ability to desegregate the biomass during the 24 h. To obtain reliable results, the test was carried out in triplicate. The data show that Tris-NAC (a formulation containing Disodium-EDTA) can disaggregate P. aeruginosa biofilm both at 8 h and at 24 h, S. aureus biofilm at 24 h (in this case at 8 h the biofilm was still absent), yet not completely disaggregate M. pachydermatis biofilm at 48 h (at 8 h and 24 h the biofilm was absent). Furthermore, the formulation inhibited the biofilm formation of all three micro-organisms.

本研究的目的是评估含有N-乙酰半胱氨酸和Tris-EDTA (TrisNAC, ICF S.r.l.;Palazzo Pignano，意大利)防止生物膜的形成和协助其解体。Clever Bioscience S.r.l.实验室(Pavia，意大利)对这种配方的生物膜生长抑制的研究，使用针对三种微生物菌株的微生物板生物膜形成试验进行了评估:铜绿假单胞菌(ATCC 27853)、金黄色葡萄球菌(ATCC 25923)和厚皮马拉色菌(DSM 6172)。同样的三种微生物在微滴皿中生长，生物膜法通过微滴皿生物膜形成试验进行评价。生物膜的生长和生产评估在两个不同的时间点(8 h, 24 h)细菌和酵母在三个不同的时间(8小时、24小时、48小时)。随后,Tris-NAC配方,在不同的稀释,添加相同的盘子评估能够废止种族隔离的生物量在24 h。获得可靠的结果,试验一式三份。数据显示,Tris-NAC(包含Disodium-EDTA配方)可以分解铜绿假单胞菌生物膜在8 h和24 h,金黄色葡萄球菌生物膜在24小时(在本例中8 h生物膜还没有),但不完全分解，厚皮马拉色菌生物膜在48小时(8 h和24小时的生物膜会消失)。此外，该配方抑制了所有三种微生物的生物膜形成。

Source of funding: ICF S.r.l. Conflicts of interest: NM is an ICF S.r.l. employee, TDM is ICF S.r.l. Scientific Manager, GG received consultancy and speaker fees from ICF S.r.l.

Ear inflammation and hearing measurements: taking advantage of preclinical tools to assess the tolerance of otic products

耳炎和听力测量:利用临床前工具评估耳部产品的耐受性

A. MARIE*, C. VINCENT*, M. DOLON† , P. SCHREIBER† , P. GRUET† , JB. RASCLE† and G. NAERT* *CILcare SAS, Montpellier, France † Virbac, Carros, France

Ear infections are a relatively common disease in pets and one of the leading causes of veterinary appointments for dogs. Therapies developed to prevent and treat otitis should be safe and well-tolerated when administered by the otic route. As some molecular agents may possess ototoxic properties and cause profound hearing impairments, their impact on ear inflammation and hearing should be assessed. The standard otoscopes and hearing tests used in veterinary clinics are insufficiently sensitive to detect slight changes in hearing and the appearance of the eardrum. In preclinical research, specific oto-endoscopes and electrophysiological measures, including brainstem auditoryevoked responses (BAER), are valuable tools to evaluate auditory changes with high precision. The BAER technique involves the detection of electrical activity of the brainstem in response to a range of frequencies sent at specific intensities. We transposed the method used widely in rodents to species such as dogs or mini swine, using a portable BAER device and an oto-endoscope equipped with a camera. The protocol was developed to detect BAER waves at different frequencies, according to the hearing range of the tested species. Oto-endoscopes specifically adapted to the anatomy of each species were used to observe the auditory canal down to the eardrum. Otoscopic and BAER evaluations were successfully developed in both species. In conclusion, a dedicated BAER device coupled to an oto-endoscope is a flexible and sensitive diagnostic tool for pets. It can be used to easily monitor ear integrity and hearing, and detect the ototoxic effects of otic products.

耳部感染在宠物中是一种相对常见的疾病，也是让犬去看兽医的主要原因之一。预防和治疗耳炎的治疗方法应该是安全的，通过耳道给药时耐受性良好。由于某些分子药物可能具有耳毒性并引起严重的听力病变，因此应评估其对耳部炎症和听力的影响。兽医诊所使用的标准耳镜和听力测试不够敏感，无法发现听力和鼓膜外观的轻微变化。在临床前研究中，特殊的耳内窥镜和电生理测量，包括脑干听觉诱发反应(BAER)，是评估听觉变化和高精度的有价值的工具。BAER技术包括检测脑干对特定强度发出的一系列频率的反应的电活动。我们使用便携式BAER设备和配备摄像机的耳内窥镜，将广泛用于啮齿类动物的方法转换为犬或迷你猪等物种。该方案是根据被测试物种的听力范围，在不同频率下检测BAER波的。专门适应于每个物种解剖的耳内窥镜被用来观察耳道到鼓膜。耳镜和BAER评估在这两个物种中都成功发展。总之，一个专用的BAER设备联合耳内窥镜是一个灵活和敏感的宠物诊断工具。它可以方便地监测耳朵完整性和听力，并检测耳毒性产品。

Source of funding: Self-funded. Conflicts of interest: None declared.

Allermmune immunotherapy in feline atopic syndrome

猫特应性综合征的过敏免疫治疗

F. MARTINI, C. FAVROT, A. ROSTAHER and N. FISCHER Clinic for Small Animal Internal Medicine, Vetsuisse Faculty, Zurich University, Zurich, Switzerland

Allermmune (Zenoaq; Fukushima, Japan) is a recombinant Dermatophagoides farinae 2 (Der f 2) pullulan-based immunotherapy vaccine whose efficacy on house dust mite (HDM)-allergic dogs has been demonstrated in several studies. The goal of the present pilot study was to evaluate the efficacy of this intervention in Der f sensitised cats. Ten affected cats diagnosed with feline atopic syndrome received Allermmune weekly for six weeks and then monthly for three months. On week (W)0, W6 and W17 clinical lesions were assessed by means of the Feline Dermatitis Extent and Severity Index (FeDESI), while owners assessed pruritus with a 10 cm Visual Analog Scale (pVAS). Concurrent medications were recorded, using a previously published scoring system (MS). Data were normally distributed, and a one-tailed paired Students’ t-test was used for statistical analysis. FeDESI, pVAS and MS means improved greatly from 37 to 12 (P = 0.003), from 7.2 to 3.2 (P = 0.001) and from 32 to 10 (P = 0.005), respectively. Only 2/10 cats were considered nonresponders. Interestingly, the improvement was already obvious (>50% improvement) at W6 in 58%, 71% and 60%, for pVAS, FeDESI and MS, respectively. Two cats with concurrent asthma and relapsing angioedema also responded adequately. The results of this study are very similar to those seen in dogs and suggest that Allermmune may be effective in alleviating clinical signs in cats with skin or respiratory signs of HDM-hypersensitivity. Further large-scale and controlled studies are needed to confirm these findings.

Allermmune (Zenoaq;是一种重组粉螨2 (Der f 2)普鲁兰免疫治疗疫苗，其对屋尘螨(HDM)过敏犬的疗效已在多项研究中得到证实。目前初步研究的目的是评估这种干预对Der f致敏猫的疗效。10只被诊断为特应性综合症的患猫接受过敏免疫治疗，为期6周，然后每月一次，持续3个月。在第0周(W)、第6周和第17周，采用猫皮炎严重程度指数(FeDESI)评估临床病变，宠主用10cm视觉模拟量表(pVAS)评估瘙痒。使用之前发表的评分系统(MS)记录并发药物。数据为正态分布，采用单尾配对student t检验进行统计分析。FeDESI、pVAS、MS平均值分别由37 ~ 12 (P = 0.003)、7.2 ~ 3.2 (P = 0.001)、32 ~ 10 (P = 0.005)显著改善。只有2/10的猫被认为无反应。有趣的是，pVAS、FeDESI和MS病患W6的改善已经很明显(改善50%)，分别为58%、71%和60%。同时患有哮喘和复发性血管性水肿的两只猫也有充分的反应。这项研究的结果与在犬身上看到的结果非常相似，并表明过敏免疫可能有效地缓解猫的皮肤或呼吸道症状的HDM超敏反应。需要进一步的大规模对照研究来证实这些发现。

Source of funding: Allermmune was provided by Zenoaq. Conflicts of interest: CF is scientific consultant for Zenoaq.

Alopecia areata concurrent with atopic dermatitis in three dogs

3只犬斑秃伴发特应性皮炎

G.A. GRAPI*, E.R. RIERA*, C. YOTTI† , J. DECLERCQ‡ and L. FERRER§ *Citopat Veterinaria and Letipharma, Barcelona, Spain † Centro Dermatologico Skinpet, Madrid, Spain  ‡ Small Animal Practice, Kortrijk, Belgium § Universitat Autonoma de Barcelona, Barcelona, Spain  

Alopecia areata is an uncommon autoimmune skin disease of the dog characterised clinically by noninflammatory alopecia and histologically by a lymphocytic bulbitis. Atopic dermatitis is a common, genetically predisposed inflammatory and pruritic skin disease associated with well-defined clinical signs and immunoglobulin (Ig)E directed against environmental allergens. We present three dogs (two chow chows, one shih tzu) with atopic dermatitis (AD) that developed alopecia areata. All three dogs had a clinical history of nonseasonal pruritus and after ruling out other pruritic diseases by appropriate diagnostic tests, a diagnosis of AD was made. The dogs responded totally or partially to oclacitinib and/or ciclosporin. Concurrently, the animals developed atypical areas of noninflammatory alopecia that were not self-induced. Histologically, the lesions were characteristic of alopecia areata and consisted of lymphocytic inflammation of the hair bulbs of anagen follicles and surrounding adjacent tissue, with scant macrophages. Additional findings were infundibular keratosis, follicular atrophy and marked vacuolisation along the follicle walls (vacuolar degeneration) confirming the diagnosis of alopecia areata. Lymphocytic infiltrates were positive to CD3, with no positivity for S100. Comorbidity between immune-mediated diseases is a wellknown phenomenon in humans and dogs. We describe for the first time the association between AD and alopecia areata, with additional new findings not described previously including intense vacuolar degeneration of follicular walls. All three dogs had a plush hair coat (chow chow, shih tzu). One of the dogs showed recurrence of hair loss during the following two years, suggesting a seasonal course.

斑秃是一种罕见的犬自身免疫性皮肤病，临床表现为非炎症性脱毛，组织学表现为淋巴细胞性毛球炎。特应性皮炎是一种常见的遗传性炎症性瘙痒性皮肤病，具有明确的临床体征和针对环境过敏原的免疫球蛋白(Ig)E。我们报告了三只犬(两只松狮犬，一只西施犬)患有特应性皮炎(AD)，并发展为斑秃。3只犬均有非季节性瘙痒史，经适当的诊断试验排除其他瘙痒病后，诊断为AD。犬对奥拉替尼和/或环孢素完全或部分有效。同时，动物出现非炎症性非典型性脱毛。组织学上表现为斑秃的特征性病变，表现为毛囊及周围组织的淋巴细胞性炎症，伴少量巨噬细胞。另外还发现漏斗部角化、毛囊萎缩和沿毛囊壁的明显空泡化(空泡变性)，证实了斑秃的诊断。CD3阳性淋巴细胞浸润，S100阴性。在人类和犬免疫介导疾病并存是一个众所周知的现象。我们首次描述了AD和斑秃之间的关系，以及以前未描述的其他新发现，包括毛囊壁强烈的空泡变性。三只犬都具有蓬松被毛(松狮犬，狮子犬)。其中一只犬在接下来的两年里出现脱毛复发，表明这是一个季节性的过程。

Source of funding: Self-funded. Conflicts of interest: None declared.

Development and validation of an owner-assessed Visual Analog Scale (VAScat) for feline pruritus severity scoring

猫瘙痒严重程度评分的宠主评估视觉模拟量表(VAScat)的开发和验证

S. COLOMBO*, S. BORIO† , R. SARTORI‡ and C. SCHIEVANO§ *Servizi Dermatologici Veterinari, Legnano, Italy † Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, California, USA ‡ Servizi Dermatologici Veterinari, Milano, Italy § University of Padova, Padova, Italy

Assessment of the severity of pruritus is difficult in cats, because they manifest discomfort by overgrooming, which includes increased licking, increased scratching or both. Our objective was to develop and evaluate the effectiveness of a feline-specific pruritus scale (VAScat). The scale was designed using the combination of a double Visual Analog Scale (VAS), one for licking and one for scratching, with severity and behavioural descriptors. The highest score (VAS-max) on either VAS was taken as the pruritus score for each cat. Owners of 153 cats with skin diseases and 108 healthy cats scored their pet’s pruritus using the VAScat; 96/153 cats with skin diseases were re-evaluated after four to eight weeks of treatment. Pearson’s correlation value between VAS-licking and VASscratching scores was r = 0.26, and Cronbach’s alpha was 0.41. Both indexes indicated that the two scales measure different manifestations of pruritus and supported the use of a dual assessment system. Comparison with a clinical pruritus severity scale (0, absent; 1, mild; 2, moderate; 3, severe) suggested that VAS-licking and VAS-scratching scales taken alone are unsuitable for measuring absent to mild pruritus (grades 0–1), while VASmax is suitable (P = 0.001). VAS-licking, VAS-scratching and VAS-max were all suitable to assess higher levels of pruritus (grades 2–3, P < 0.01). The VAScat was able to measure pruritus improvement following therapy, as post-treatment scores were significantly decreased compared to pre-treatment ones (P < 0.0001). It thus proved to be a useful tool to assess pruritus in cats and for monitoring the response to treatment.

评估猫的瘙痒程度是困难的，因为它们通过过度梳理表现出不适，包括增加舔舐，增加抓挠或两者都有。我们的目的是开发和评估一种猫特异性瘙痒量表(VAScat)的有效性。该量表的设计采用了双视觉模拟量表(VAS)，一种用于舔舐，另一种用于抓挠，并包含严重程度和行为描述。以每只猫的瘙痒评分(VAS-max)为最高评分。153只患有皮肤病的猫和108只健康的猫的主人用VAScat对宠物的瘙痒程度进行了评价。在治疗4至8周后，对153只患有皮肤病的猫进行了重新评估。Pearson相关值为r = 0.26, Cronbach s α为0.41。这两个指标表明，这两个量表测量不同的瘙痒表现，并支持使用双重评估系统。与临床瘙痒严重程度量表(0，无;1、温和;2、温和;3，严重)提示仅取vas -舔舐和vas -挠痒量表不适用于无瘙痒至轻度瘙痒(0级1)，而VASmax适合(P = 0.001)。vas -舔痒、vas -抓痒和VAS-max均适用于评估瘙痒程度较高的病患(2 - 3级，P <0.01)。VAScat能够测量治疗后瘙痒改善情况，因为治疗后评分与治疗前相比显著降低(P <0.0001)。因此，它被证明是一个有用的工具，以评估瘙痒在猫和监测对治疗的反应。

Source of funding: SIDEV Research Grant. Conflicts of interest: None declared.

Does a disease resembling human psoriasis exist in dogs?

犬身上是否存在一种类似人类银屑病的疾病?

ORDEIX*, A. FONDATI† , F. SCARAMPELLA‡ and C. MARIA§ *Fundacio Hospital Cl  ınic Veterinari, Barcelona, Spain † Veterinaria Trastevere and Veterinaria Cetego, Rome, Italy ‡ Studio Dermatologico Veterinario, Milan, Italy § Servei de Dermatologia, Fundacio Hospital Cl  ınic Veterinari, Barcelona, Spain

Human psoriasis is an immune-mediated disease resulting from gene–environmental trigger interactions, characterised clinically by chronic, symmetrical, welldemarcated scaling erythematous plaques, and histologically by acanthosis, parakeratosis and lymphocytic infiltrate. We report four dogs with clinicopathological findings resembling human psoriasis. Two spayed female Belgian shepherd dogs and two male Jack Russell terriers with a median age of 7 years (range 2–8) were presented with a chronic (9 month history; 6–30) and intensely pruritic dermatitis. Predominant lesions were symmetrical erythematous plaques covered by whitish adherent scales on the extremities and bony prominences (n = 4), periocular (n = 4) and perioral (n = 3) regions, inner pinnae (n = 3), footpads (n = 3) and scrotum (n = 1). Histopathological lesions were regular acanthosis with parakeratosis, spongiosis, lymphocytic exocytosis, lymphocytic perivascular dermatitis (n = 4) and intraepidermal neutrophilic micropustules (n = 2). Three dogs exhibited lack of remission following elimination diet and allergen-specific serological results were negative (n = 1) and positive for salivary flea (n = 1) and Ambrosia (n=1). Oral glucocorticoids led to partial improvement in three dogs, although relapses following treatment tapering were observed. Pruritus, and not lesions, partially responded to a six month course of oclacitinib in one dog. Complete clinical response (n = 4) was achieved with 5 mg/kg ciclosporin (41 days; range 10–70). Median follow-up was 8.5 months. One dog showed a mild relapse when ciclosporin was administered once weekly and another dog did not show any relapse one year after ciclosporin discontinuation. Chronic erythematous-scaling dermatitis associated with hyperplastic and parakeratotic lymphocytic dermatitis may represent a psoriasis-like distinctive clinicohistopathological reaction pattern related to an immune-mediated response whose triggers remain currently unknown.

人类银屑病是一种由基因环境触发相互作用引起的免疫介导疾病，临床表现为慢性、对称、边界清楚的皮屑性红斑斑块，组织学表现为棘层症、角化不全和淋巴细胞浸润。我们报告四个犬与临床病理发现类似人银屑病。2只绝育的雌性比利时牧羊犬和2只雄性杰克罗素㹴，中位年龄为7岁(范围为28岁)，有慢性(9个月的病史;6-30)和严重瘙痒性皮炎。主要病变是对称的红斑的斑块被白色皮屑覆盖，位于四肢和骨突(n = 4),眼周(n = 4)和口周(n = 3)区域,耳廓凹面(n = 3),爪垫(n = 3)和阴囊(n = 1)。组织病理学病变是常规棘皮症与角化不全、海绵层水肿,淋巴细胞外泌、淋巴细胞血管周皮炎(n= 4)和表皮内中性粒细胞小脓疱(n= 2)。3只犬在食物排查后出现症状缓解，过敏原特异性血清学结果为阴性(n=1)，跳蚤唾液(n=1)和豚草 (n=1)呈阳性。口服糖皮质激素3只犬得到部分改善，但在治疗剂量逐渐减量后复发。在一只犬身上进行了6个月疗程的奥拉替尼治疗后，出现部分缓解，有瘙痒但没有皮肤病变。使用5mg/kg环孢素临床完全缓解(n = 4)(41天;范围10-70)。平均随访时间为8.5个月。当环孢素每周给药一次时，其中一只犬出现了轻微的复发，而另一只犬在停用环孢素一年后没有出现任何复发。与增生性和角化不全性淋巴细胞性皮炎相关的慢性红斑-皮屑性皮炎可能代表一种银屑病样独特的临床组织病理反应模式，该反应与免疫介导反应相关，其触发机制目前尚不清楚。

Source of funding: Self-funded. Conflicts of interest: None declared.

Dogs with moderate to severe leishmaniosis present an exaggerated allergen-specific immunoglobulin E immune response

中度至重度利什曼病犬表现出夸张的过敏原特异性免疫球蛋白E免疫反应

M. CABRE*, L. SOLANO-GALLEGO† and L. ORDEIX* *Fundacio Hospital Cl  ınic Veterinari, † Departament de Medicina i Cirurgia Animals, Universitat Autonoma de  Barcelona, Barcelona, Spain

Dogs with moderate to severe canine leishmaniosis develop a strong nonprotective humoral response which is associated mainly with a massive production of  immunoglobulin (Ig)G, although IgE also is produced. The aim of this study was to determine the allergen-specific IgE response in dogs with moderate to severe leishmaniosis and to compare it with that observed in clinically healthy dogs. Serum samples from dogs with leishmaniosis (n = 26) without atopic dermatitis and with high L. infantum-specific antibody levels were studied by means of a commercially available allergen-specific IgE assay kit (Polycheck Canis 20, Biocheck GmbH; Munster, Germany); serums from clinically healthy dogs (n = 21) negative (＜35 ELISA units) to the same L. infantum in-house ELISA served as controls. The median ages of dogs with leishmaniosis and clinically healthy dogs were 4 (range 1– 10) and 6 years (range 1–13), respectively (Wilcoxon– Mann–Whitney U-test, P = 0.129). IgE anti-cross-reactive carbohydrate determinants (CCD) were detected in 4/26 (15.4%) dogs with leishmaniosis and 3/21 (14.3%) clinically healthy dogs (Fisher’s exact test, P = 0.916). Therefore, only dogs without anti-CCD IgE were analysed. Total allergen-specific IgE concentration was higher in dogs with leishmaniosis (mean 27.28 kU/L) compared to healthy dogs (mean 14.06 kU/L) (Wilcoxon–Mann–Whitney U-test, P = 0.015). Specific-IgE-positive results against at least one allergen were detected in 21/22 (95.5%) dogs with leishmaniosis and in 12/18 (66.7%) clinically healthy dogs (chi-square, P = 0.017). This preliminary study indicates that dogs with moderate to severe leishmaniosis present an exaggerated allergenspecific IgE immune response that does not appear to be of clinical relevance.

患有中度至重度犬利什曼病的犬会产生强烈的非保护性体液反应，这主要与大量产生免疫球蛋白(Ig)G有关，尽管也会产生IgE。本研究的目的是确定中度至重度利什曼病犬的过敏原特异性IgE反应，并将其与临床健康犬的观察结果进行比较。用一种市售的过敏原特异性IgE检测试剂盒;从临床健康犬(n = 21)的血清(<35个ELISA单位)到相同的L.婴儿的内部ELISA作为对照。利什曼病犬和临床健康犬的中位年龄分别为4岁(范围1 10)和6岁(范围1 13)。在4/26(15.4%)利什曼病犬和3/21(14.3%)临床健康犬中检测到IgE抗交叉反应碳水化合物(CCD) (Fisher精确试验，P = 0.916)。因此，只对无抗ccd IgE的犬进行分析。利什曼病犬总过敏原特异性IgE浓度(平均27.28 kU/L)高于健康犬(平均14.06 kU/L) 。在21/22(95.5%)利什曼病犬和12/18(66.7%)临床健康犬中检测到至少一种过敏原特异性ige阳性(卡方，P = 0.017)。本初步研究表明，中度至重度利什曼病犬存在过度的过敏原特异性IgE免疫反应，这似乎没有临床意义。

Source of funding: 2019 ESVD Minor Grant. Conflicts of interest: None declared.

Effect of a natural spot-on based on phytoceramides, plant-extracted essential fatty acids and essential oils on reconstructed canine epidermis

基于植物神经酰胺、植物提取的必需脂肪酸和精油的天然滴剂对重建犬表皮的影响

C. DARMON-HADJAJE*, J. DELLACASAGRANDE† and N. AMALRIC† *CHV Fregis, Arcueil, France  † Synelvia, Labege, France  

Atopic dermatitis (AD) involves cutaneous barrier impairment linked to inflammation. Reconstructed canine epidermis (RCE) from healthy dog keratinocytes cultured with T-helper (Th)1 and Th2 cytokines for four days (RCEAD) mimics changes observed in canine AD. This study evaluated the effect of a natural complex designed specifically for allergy-prone skin contained in a spot-on [Dermoscent ATOP 7, Laboratoire de Dermo-Cosmetique  Animale (LDCA); Castres, France] in RCE-AD model. Three RCE-ADs were treated with 0.02% ATOP 7 spoton complex for four days; three RCE-AD and three normal RCEs treated with a placebo served as controls. Qualitative histological analysis evaluated RCE morphology, whereas profilaggrin and filaggrin expressions were quantified through immunofluorescence staining using IMAGEJ software. Inflammation was assessed by measuring the secreted canine cytokine interleukin (IL)-8 by enzymelinked immunosorbent assay. Results are expressed as mean  SD. After having confirmed the good tolerance of the natural complex on normal RCEs, the efficacy of the natural complex then was tested on RCE-AD. Morphological disorganisation was observed on control RCE-AD, while improvement with less spongiosis was noted after treatment. Immunofluorescence staining revealed a 52% improvement in profilaggrin and filaggrin expressions comparing the treated RCE-AD quantified at 32.58%  6.3 and the control RCE-AD at 21.47%  7.79, both compared to normal RCEs as baseline (100%). RCE-AD treated with the natural complex showed 25% reduction in IL-8 release (30,423 pg/ mL  6,597) compared to the control RCE-AD at 40,471 pg/mL  4,110. ATOP 7 spot-on could be of interest in the multimodal management of canine AD thanks to its natural complex demonstrated to decrease IL-8 and improve skin barrier function of RCE-AD models.

特应性皮炎(AD)涉及与炎症相关的皮肤屏障损害。用T 辅助 (Th)1和Th2细胞因子培养健康犬角质细胞(RCEAD)，构建的犬表皮细胞(RCE)模拟了犬AD的变化。本研究评估了一种专为易过敏皮肤设计的天然复合物的效果，该复合物包含在一种滴剂中[德美丝 ATOP 7, Laboratoire de dermo - cosmetics  Animale (LDCA);Castres，法国]在RCE-AD模型中。0.02% ATOP 7 滴剂在复合物上处理3个RCE-ADs四天;3个RCE-AD和3个正常的RCE与安慰剂一起作为对照。定性组织学分析评价RCE的形态，而前丝聚蛋白和丝聚蛋白表达的全通过免疫荧光染色，使用IMAGEJ软件。通过酶联免疫吸附法检测犬白细胞介素(IL)-8的分泌情况。结果用平均SD表示。在确认天然配合物对正常RCE具有良好耐受性后，再对RCE-AD进行药效测试。对照组RCE-AD观察到形态紊乱，治疗后改善，海绵状病变减少。免疫荧光染色显示，与正常RCE(baseline, 100%)相比，处理RCE-AD (32.58%  6.3)和对照RCE-AD(21.47% 7.79)的前丝聚蛋白和丝聚蛋白表达均有52%的改善。IL-8释放量(30,423 pg/mL 6,597)比对照组(40,471 pg/mL 4,110)降低了25%。由于其天然复合物被证明可以降低IL-8并改善RCE-AD模型的皮肤屏障功能，在犬AD的多模式管理中，ATOP 7滴剂可能有用。

Source of funding: LDCA. Conflicts of interest: C. Darmon is a consultant for LDCA.

Skin barrier reinforcement effect of a spot-on based on natural ingredients in a dog model of tape-stripping

一种天然成分滴剂在犬胶带粘贴模型上的强化皮肤屏障作用

A. IDEE, M. MOSCA and D. PIN Vetagrosup, Marcy l’etoile, France  

Canine atopic dermatitis being linked to skin inflammation and barrier alterations, cutaneous barrier reinforcement is therefore essential in a multimodal approach. This study aims to evaluate the barrier reinforcement effect of a spot-on [Dermoscent ATOP 7, Laboratoire de DermoCosmetique Animale (LDCA); Castres, France], with a  noninvasive method of acute barrier disruption by stratum corneum delamination with repeated tape-stripping. Five healthy adult beagle dogs were enrolled in this study. A fixed number of adhesive tapes were applied on each of four predesignated clipped zones (Z1–4) on the back, at one zone per week. The spot-on was applied weekly for three weeks on the same spot, at the medium point of the four zones. Its effect on transepidermal water loss (TEWL) was measured (AF200 AquaFlux, Biox Systems; London, UK) weekly following delamination on Z2, Z3, Z4, four days after each spot-on application. Z1 (before first application of the spot-on) and a nondelaminated zone on the abdomen were used as controls. Tape-stripping induced a significant increase in TEWL on Z1 (before first spot-on application) compared to nondelaminated abdomen zone (77.62 17.68 versus 20.46 10.24 g/m2 /h, P < 0.001, ANOVA2 test). Spot-on applications progressively limited the TEWL after delamination, with a TEWL close to that of the abdomen area after the 3rd application (40.56  17.51 versus 21.64  10.51 g/m2 /h, no significant difference, ANOVA2 test). Moreover, the spot-on demonstrated its diffusion efficacy at distance from the application point. No deleterious effect was noted. Weekly application of this spot-on at one point during three weeks protects and strengthens the skin by limiting TEWL in case of barrier alteration.

犬特应性皮炎与皮肤炎症和屏障改变有关，因此皮肤屏障强化在多模式方法中至关重要。本研究旨在通过反复胶带黏贴使角质层的层离，造成屏障破坏的无创方法，评价滴剂[Dermoscent ATOP 7, Laboratoire de DermoCosmetique Animale (LDCA); Castres, France]对屏障的加强作用。。5只健康的成年比格犬参与了这项研究。在背部的四个预先指定的剃毛区域(Z1-4)中，每个区域应用固定数量的胶带，每周一个区域。在四个区域的中心点，每周用一次滴剂，连续三周。每次用滴剂后4天，每周测量（AF200 AquaFlux, bix Systems;伦敦，英国)其对Z2、Z3、Z4层离的经表皮失水(TEWL)的影响。Z1(第一次滴剂使用前)和腹部的非层离区域作为对照。与腹部非层离区域相比，Z1组(第一次滴剂使用前)的皮肤水分流失显著增加(77.62、17.68 vs 20.46、10.24 g/m2 /h, P <0.001, ANOVA2测试)。滴剂的应用逐步限制层离后的水分流失，第3次滴剂应用后的水分流失接近腹部区域(40.56  17.51 g/m2 /h vs 21.64  10.51 g/m2 /h, ANOVA2检验无显著差异)。此外，在距离滴点较远的地方，该滴剂的扩散效果显著。没有发现有害影响。在三周的时间里，每周在一点上涂滴剂，防止皮肤屏障改变，减少水分流失，从而保护和加强皮肤。

Source of funding: The study was funded by LDCA. Conflicts of interest: None declared.

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