Co-occurrence of papillomas related to Equus caballus papillomavirus type 2 and cutaneous habronemiasis

The present report describes a case of cutaneous papillomatosis related to equine papillomavirus type 2 (EcPV2) with concomitant cutaneous habronemiasis. A 2-year-old female Pura Raza Española horse was referred for a dermatological evaluation because of the onset of cutaneous multifocal, nodular lesions on the face characterised by a warty and partially ulcerated overlying epidermis. Multiple biopsies were taken from these lesions, and cutaneous viral papillomas with a concomitant severe focal eosinophilic dermatitis with intralesional Habronematidae nematode larvae were diagnosed. Immunohistochemistry revealed papillomaviral capsid protein L1 in the epidermal proliferative lesion, confirming the presence of a productive infection. Rt-qPCR confirmed the presence of EcPV type 2. To the authors’ knowledge, this is the first report of EcPV2-related papillomas with concurrent habronemiasis.     

Variations in the morphology of Rhizomucor pusillus in granulomatous lesions of a Magellanic penguin (Spheniscus magellanicus)

This report presents a new case of mucormycosis encountered in penguin characterized by morphological variation of hyphae and presence of sporangia with numerous sporangiospores. A 4.5-year-old Magellanic penguin (Spheniscus magellanicus) died after exhibiting anorexia, poor nutritional condition and dyspnea. Multiple nodular lesions were observed in the thoracic and abdominal regions. Histopathologically, hyphae of various sizes were seen in the lungs, air sac and nodular lesions. Myriad sporangiospores and several sporangia were observed in/around the bronchi or parabronchi. The very narrow and short hyphae in the nodules were not consistent with the characteristics of Mucorales. However, for most hyphae, including those in the nodules, sporangiospores and sporangia, immunohistochemistry revealed Mucorales-positive reactions. In addition, these fungi were identified as Rhizomucor pusillus by gene analysis.     

Analysis of genomic mutation and immunohistochemistry of platelet‐derived growth factor receptors in canine vascular tumours

We examined whether mutation of the platelet‐derived growth factor receptor protein tyrosine kinase (PDGFR)‐α and PDGFR‐β genes contributes to their overexpression in canine vascular tumours. Genomic sequences of trans‐ or juxtamembrane regions of PDGFR‐α and PDGFR‐β were analysed with immunohistochemical staining and polymerase chain reaction‐direct sequencing using DNA from paraffin‐embedded neoplastic tissues of 27 hemangiosarcomas (HSAs) and 20 hemangiomas (HAs). Immunohistochemically, 75% of the HA cases were positive for PDGFR‐α and almost most of the HA cases were negative for PDGFR‐β. Of the HSA cases, 55.6% were negative for PDGFR‐α and 63% were strongly positive for PDGFR‐β. Among the HA cases, 1 missense mutation was detected in PDGFR‐α exon 18 and 1 in PDGFR‐β exon 17. Two HSA cases had missense mutations in exon 14 and 1 in exon 17 of PDGFR‐β. Thus, genomic mutation of trans‐ or juxtamembrane regions of PDGFRs was not the main mechanism driving the activation of receptors in HSA and HA.     

The value of molecular expression of KIT and KIT ligand analysed using real‐time polymerase chain reaction and immunohistochemistry as a prognostic indicator for canine cutaneous mast cell tumours

This study investigated the correlation between KIT gene expression determined by immunohistochemistry and real‐time polymerase chain reaction (RT‐PCR) and the rate of tumour recurrence and tumour‐related deaths in dogs affected with mast cell tumour (MCT). Kaplan–Meier curves were constructed to compare tumour recurrence and tumour‐related death between patients. The log‐rank test was used to check for significant differences between curves. KIT‐I, KIT‐II and KIT‐III staining patterns were observed in 9 (11.11%), 50 (61.73%) and 22 (27.16%) tumours, respectively. Tumour recurrence rates and tumour‐related deaths were not associated with KIT staining patterns (P = 0278, P > 0.05), KIT (P = 0.289, P > 0.05) or KIT ligand (P = 0.106, P > 0.05) gene expression. Despite the lack of association between KIT staining pattern and patient survival time, the results suggest a correlation between aberrant KIT localization and increased proliferative activity of MCTs. RT‐PCR seems to be a sensible method for quantitative detection of KIT gene expression in canine MCT, although expressions levels are not correlated with prognosis.     

After nasopharyngeal infection,foot-and-mouth disease virus serotype A RNA is shed in bovine milk without associated mastitis

Foot-and-mouth disease (FMD) is a highly contagious aphthoviral infection of cloven-hoofed animals, inducing vesiculopustular stomatitis, pododermatitis, and thelitis. Vesicular fluid represents a major pathway of virus excretion, but bovine milk is another important source of virus shedding. We describe here the time course of FMD virus (FMDV) excretion in the milk and characterize associated lesions in the mammary gland. Three dairy cows were infected by nasopharyngeal instillation of FMDV and monitored over 12 d. Autopsy was performed at the end of the study, and specimens were collected for histopathology, IHC, and RT-qPCR. All 3 cows developed fever, drooling, vesiculopustular stomatitis, interdigital dermatitis, and thelitis. FMDV RNA was detectable in whole milk until the end of the trial, but only transiently in saliva, nasal secretions, and blood serum. Although histology confirmed vesiculopustular lesions in the oral and epidermal specimens, the mammary glands did not have unequivocal evidence of FMDV-induced inflammation. FMDV antigen was detectable in skin and oral mucosa, but not in the mammary gland, and FMDV RNA was detectable in 9 of 29 samples of squamous epithelia but only in 1 of 12 samples of mammary gland.     

Oncology Virtual Issue Articles

The following two articles have been published online as part of the EVE Oncology Virtual Issue which is available online at wileyonlinelibrary.com/journal/eve     

黄鳝胃肠的生长抑素分泌细胞及超微结构

在肠卷石蜡法切片上，用ABC免疫组织化学方法，显示消化管全长的生长抑素分泌细胞（又称D细胞），结果只在胃体部显现，主要散在分布于胃腺中，个别出现在粘膜上皮。消化道内分泌细胞根据基部有否胞质突起，可分为4种类型，在黄鳝胃中均有，其中Ⅲ型数量最多，I、Ⅳ型次之，而Ⅱ型数量最少。由于产生生长抑素的D细胞在消化道内分泌细胞中作用多并且类型复杂，故黄鳝胃还是一个十分重要且复杂的内分泌器官。应用透射电镜，对黄鳝的胃肠作超薄切片观察显示在胃腺上皮细胞内含大量的微管泡系和一定量的酶原颗粒，表明其兼有泌酸及产生酶原的功能，属泌酸胃酶细胞。在肠上皮细胞游离面，含有大量微绒毛，但幼鳝比成鳝的长、密且整齐，提示幼鳝的肠上皮比成鳝有更强的吸收能力。     

栉孔扇贝神经节一氧化氮合酶的组织化学和免疫组化定位

采用组织化学和免疫组化技术对栉孔扇贝(Chlamys farreri)神经节内的一氧化氮合酶(NOS)进行定位研究。组织化学显示,存在NOS的部位如下：脑神经节内纵行的神经纤维和表层的少量小细胞;足神经节表层的大量小细胞,中央大量水平分布的神经纤维;脏神经节中部大量水平分布的神经纤维,前叶内大量小细胞和神经纤维,后叶内少量小细胞和许多环行神经纤维,侧叶内大量似放射状分布的神经纤维;脑足和脑脏神经索内的神经纤维。免疫组化定位表明,神经型一氧化氮合酶(nNOS)和诱导型一氧化氮合酶(iNOS)在整个神经系统内均呈阴性;足神经节和脏神经节内有少量神经细胞呈内皮型一氧化氮合酶(eNOS)强阳性;各神经节和神经索内的部分小细胞和神经纤维呈eNOS弱阳性。栉孔扇贝进化上为较低等的贝类,NOS阳性神经细胞应主要分布于外周器官组织内。神经系统内大量的NOS可在其神经传导和免疫调节等方面发挥重要的作用。     

The GnRH systems in the brain and pituitary of normal and hCG treated European silver eels

The distribution of immunoreactive GnRH was studied in the brain and pituitary gland of normal and human chorionic gonadotrophin (hCG) injected silver eels. It was found that the general organization of GnRH systems in this species is similar to that reported in other teleosts. Cell bodies were present in the olfactory bulbs, ventral telencephalon, periventricular hypothalamus and dorsal tegmentum. No positive perikarya could be detected in the preoptic region. Only scarce fibers were observed in the proximal neurohypophysis. Treatment with hCG does not modify the distribution of GnRH but it increases the density of positive structures, in particular at the level of the pituitary. The results are discussed in relation with the present status of knowledge of the mechanisms underlying the blockage of sexual maturation in the European eel at the silver stage.     

An alkaline phosphatase-based method for the detection of infectious salmon anaemia virus (ISAV) in tissue culture and tissue imprints

Currently, the presence of infectious salmon anaemia virus (ISAV) is often detected in Atlantic salmon by the use of an indirect fluorescent antibody test. This test is limited by the poor stability of fluorescein isothiocyanate which fades after about a week in storage, preventing the development of stained archive material as a reference source. One possible alternative would be the use of immunohistochemical staining methods to detect ISAV. An immunohistochemical method is presented that uses alkaline phosphatase‐conjugated antibodies and Vector^® Red as a substrate, to detect ISAV in kidney imprints. This paper also describes a procedure where Bouin's fluid is used to successfully inhibit endogenous alkaline phosphatase in tissue samples, prior to immunohistochemical processing. This method provides a stable stain that can be read for many weeks after staining or archived for future reference.