Dirofilaria repens—An etiological factor or an incidental finding in cytologic and histopathologic biopsies from dogs

Dirofilaria repens is an endemic, zoonotic parasite of carnivores, causing subcutaneous dirofilariasis, which is mostly asymptomatic. The aim of this study was to describe 22 cases of canine subcutaneous dirofilariasis. The cytologic and histopathologic samples were collected from dogs, which presented with various clinical signs such as cutaneous/subcutaneous nodules, hydropericardium, ascites, and lymphadenomegaly. The cytologic or histopathologic examination revealed purulent, pyogranulomatous, granulomatous or eosinophilic dermatitis/panniculitis, and the presence of D repens microfilariae or adults. The microfilariae or adults were also found incidentally in neoplastic cutaneous or subcutaneous tumors and in a sialocele. For the first time, microfilariae were also detected and described in pericardial and abdominal effusions and in enlarged reactive lymph nodes. Although it is well known that D repens can cause dermatitis and panniculitis in dogs, no previous reports of D repens microfilariae in body cavity fluids were found. The role of this parasite in the accumulation of body cavity fluid or in reactive lymphadenomegaly requires further investigation. Due to its zoonotic potential, and apparently underestimated pathogenicity, each case of canine subcutaneous dirofilariasis should be treated.     

Epidermal dysplasia and Malassezia infection in two West Highland White Terrier siblings: an inherited skin disorder or reaction to severe Malassezia infection?

Two 9-month-old West Highland White Terrier siblings were referred to our clinic with pruritus, alopecia and lichenification. Cytological examination of Scotch tape strippings revealed Malassezia organisms and cocci. Skin biopsy specimens showed epidermal dysplasia. Treatment included bathing with a 2% miconazole/chlorhexidine-containing shampoo, orally administered ketoconazole (5 mg kg-1, every 12 h) and cloxacillin (25 mg kg-1 every 8 h). Six weeks later, the dermal infection had resolved and there was hair regrowth. However, the dogs were still moderately pruritic. Intradermal allergy testing was positive for house dust mites, storage mites and Malassezia. Immunotherapy was initiated, and treatment with ketoconazole and cloxacillin was stopped. Skin biopsies, which were performed in both dogs 4 months after the first presentation, revealed mild superficial perivascular dermatitis. The remaining mild facial pruritus was easily controlled with topical treatment. These two cases indicate that epidermal dysplasia might be an inflammatory or hypersensitivity reaction to the Malassezia infection or a result of excessive self-trauma, rather than a congenital keratinization disorder.     

Dermal mass aspirate from a Persian cat

A 1-year-old spayed female Persian cat with alopecia and weight loss had numerous variably ulcerated dermal nodules. Cytologic examination of an aspirate of one of the nodules revealed pyogranulomatous inflammation along with septate hyphae and basophilic round bodies, 0.5–1.0 μm in diameter, surrounded by a thin clear halo (arthrospores). The cytologic diagnosis was dermatophytic pseudomycetoma. Histologically, there were dermal granulomas containing poorly staining, septate hyphae with bulbous spores embedded within abundant amorphous eosinophilic material (Splendore-Hoeppli reaction), and the histologic diagnosis was pseudomycetoma-associated chronic multifocal severe granulomatous dermatitis with lymphocytic perifolliculitis and furunculosis. Microsporum canis was cultured from the lesion. Pseudomycetomas are distinguished from fungal mycetomas, or eumycotic mycetomas, by the findings of multiple lesions, lack of a history of skin trauma, an association with dermatophytes, most commonly Microsporum canis , and, histologically, lack of true cement material and a more abundant Splendore-Hoeppli reaction in pseudomycetomas. Additionally, pseudomycetomas differ from dermatophytosis, in which lesions are restricted to epidermal structures. Persian cats have a high incidence of pseudomycetoma formation, suggesting a heritable predisposition. The prognosis is fair with systemic antifungal therapy. When examining cytologic specimens from Persian cats with single or multiple dermal nodules, especially if pyogranulomatous inflammation is present, a diagnosis of pseudomycetoma should be suspected and is warranted if arthrospores and refractile septate hyphae are present.     

Acellular Dermal Matrix for Surgical Repair of Ventral Hernia in Horses

The aim of the present study was to evaluate acellular dermal matrix (ADM) of rat origin for the repair of ventral hernia in horses. The skin from rats, to be used as a graft, was de-epithelialized using hypertonic solution and further decellularized with 0.5% sodium dodecyl sulfate and 0.25% tri-(n-butyl)-phosphate. Under general anesthesia, the hernial ring was exposed and repaired with the ADM graft using inlay graft technique. Blood samples were collected at postimplantation days 0, 15, and 30 and were used for sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis to assess the serum protein concentration of the animals, as well as for gelatin zymography for the identification of matrix metalloproteinases. All animals had an uneventful recovery without clinical signs of wound dehiscence, infection, or recurrence of hernias during 6-month follow-up period. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of concentration of the serum proteins revealed that this was increased at day 15 and had decreased again at day 30. Gelatin zymography showed only one major band of 92 kDa in the serum of all the horses with the implant, but the relative amount of 92 kDa was higher at day 15 as compared with day 0 and day 30. It may be concluded that ADM of rat origin can be used safely in horses for repair of ventral hernia.     

Surgical treatment of a large congenital cavernous haemangioma on the thorax of a foal

A 3‐day‐old male foal was presented with a fluctuant 25 × 15 cm mass on the thorax. The mass had increased in size since birth. The mass did not respond to conservative treatment consisting of aspiration of serohaemorrhagic contents and compression bandages, and it was therefore surgically removed when the foal was approximately 2½ weeks. A histopathological diagnosis of cavernous haemangioma was made. Healing progressed without complications despite a large surgical wound left to heal by second intention. Recurrence was not observed during the 1.5‐year follow‐up period.     

Unusual Glycosaminoglycans from a Deep Sea Hydrothermal Bacterium Improve Fibrillar Collagen Structuring and Fibroblast Activities in Engineered Connective Tissues

Biopolymers produced by marine organisms can offer useful tools for regenerative medicine. Particularly, HE800 exopolysaccharide (HE800 EPS) secreted by a deep-sea hydrothermal bacterium displays an interesting glycosaminoglycan-like feature resembling hyaluronan. Previous studies demonstrated its effectiveness to enhance in vivo bone regeneration and to support osteoblastic cell metabolism in culture. Thus, in order to assess the usefulness of this high-molecular weight polymer in tissue engineering and tissue repair, in vitro reconstructed connective tissues containing HE800 EPS were performed. We showed that this polysaccharide promotes both collagen structuring and extracellular matrix settle by dermal fibroblasts. Furthermore, from the native HE800 EPS, a low-molecular weight sulfated derivative (HE800 DROS) displaying chemical analogy with heparan-sulfate, was designed. Thus, it was demonstrated that HE800 DROS mimics some properties of heparan-sulfate, such as promotion of fibroblast proliferation and inhibition of matrix metalloproteinase (MMP) secretion. Therefore, we suggest that the HE800EPS family can be considered as an innovative biotechnological source of glycosaminoglycan-like compounds useful to design biomaterials and drugs for tissue engineering and repair.     

白蜡及白蜡高级烷醇对人真皮乳头细胞生长的影响

[目的]明确白蜡及白蜡高级烷醇吐温水溶液对人真皮乳头细胞(HFDPCs)生长的影响。[方法]采用台盼蓝法测定细胞的的接种数量,接种第2天给药,给药48 h后采用MTS法研究不同浓度吐温80、白蜡及白蜡高级烷醇吐温水溶液对人真皮乳头细胞的作用。[结果]人真皮乳头细胞在吐温80用量1.56 6.25μg·m L~(-1)范围内能正常生长。在安全范围内,白蜡(增溶量为0.31 1.25μg·m L~(-1))吐温水溶液、白蜡高级烷醇(增溶量为3.12 12.5μg·m L~(-1))吐温水溶液均能促进人真皮乳头细胞的增殖,其中,水溶液中含有6.25μg·m L~(-1)吐温80和1.25μg·m L~(-1)白蜡、水溶液中含有6.25μg·m L~(-1)吐温80和12.5μg·m L~(-1)白蜡高级烷醇时细胞的增殖率明显高于非那雄胺。[结论]吐温80作为增溶剂,其用量低于6.25μg·m L~(-1)时对人真皮乳头细胞安全。在保证吐温80对细胞安全的条件下,白蜡及白蜡高级烷醇可显著促进人真皮乳头细胞的增殖。     

施药条件对施药者体表污染的影响

该文主要研究了在夏季高温季节使用3WS-16型手动喷雾器和WFB-18AC型机动喷雾器施药对施药者体表的农药污染情况及影响施药者体表污染的主要施药条件。结果表明：使用手动喷雾器,左手、右手、右上臂和右前臂污染较多;使用机动喷雾器,左上臂和左前臂污染较多;施药者体表主要污染部位与次要污染部位间具有高度的线性相关;施药机具和喷头流量、作业环境、作物因素和施药方式等单因素均影响施药者体表污染;单因素交互作用后,风速和作物高度是影响施药者体表污染的主要因素。此研究结果对于改进施药技术,减少施药者体表污染具有实际的应用价值。     

大眼鰤鲈鱼真皮肿瘤病毒（WDSV）辅助基因orfA和orfC多抗的制备及初步应用

【目的】制备大眼鰤鲈鱼真皮肿瘤病毒(WDSV)辅助基因orfA和orfC的多克隆抗体,并用其检测酵母和肿瘤细胞中目的蛋白的表达。【方法】以WDSV基因组序列为模板,PCR扩增WDSV orfA和orfC基因,分别构建其原核表达载体,转化Rosseta（DE3）感受态细胞,经IPTG诱导表达并纯化重组蛋白后,采用皮下多点注射结合耳静脉加强免疫的方法免疫新西兰白兔,制备抗orfA和抗orfC的多克隆抗体,用Western blot法检测抗体的特异性。用制备的多克隆抗体检测orfA和orfC融合蛋白在酵母和HeLa299、SPC-A-1肿瘤细胞中的表达。【结果】PCR扩增获得了WDSV orfA和orfC基因。成功构建了orfA和orfC基因的原核表达载体并进行了诱导表达,orfA和orfC重组融合蛋白以包涵体形式存在;采用皮下注射结合耳静脉加强免疫有效地提高了抗体效价,成功制备得到了兔抗orfA和兔抗orfC血清,抗体效价达1∶8 000～1∶10 000;通过Western blot法,利用上述抗血清分别检测到了原核表达的目的蛋白及酵母和肿瘤细胞中表达的目的蛋白。【结论】制备得到的抗orfA和抗orfC多克隆抗体具有很好的特异性,能够用于原核表达、酵母和肿瘤细胞中表达的目的蛋白的Western blot检测。     

Allele frequency of hereditary equine regional dermal asthenia in American Quarter horses in Brazil determined by quantitative real-time PCR with high resolution melting analysis

Hereditary equine regional dermal asthenia (HERDA) is a genetic disorder that occurs in the American Quarter horse (AQH) and is caused by a c.115G>A missense mutation in the peptidylprolyl isomerase B (PPIB) gene. Using a quantitative real-time PCR high resolution melting analysis genotyping assay for the PPIB mutation, the estimated HERDA allele and carrier frequencies in a sample of Brazilian AQHs were 2.9% and 5.8%, respectively.