患真菌性皮肤病奶牛血液中白细胞分类计数及酯酶染色阳性T淋巴细胞的检测与分析

为探讨T淋巴细胞在皮肤真菌病奶牛中的作用,对患皮肤真菌病的奶牛和健康对照牛静脉血中的T淋巴细胞数量进行了检测。采用酯酶染色法观察并记录静脉血中酯酶染色T淋巴细胞的数量,并用瑞氏染色对白细胞进行分类计数。结果表明,患病奶牛的T淋巴细胞在淋巴细胞中的百分比明显高于健康对照奶牛,说明T淋巴细胞在抗皮肤真菌病免疫中发挥了重要的作用。     

Dermatophytosis due to Microsporum persicolor (13 cases) or Microsporum gypseum (20 cases) in dogs

A retrospective study of 20 cases of dermatophytosis due to Microsporum gypseum and 13 cases due to Microsporum persicolor seen over an 8-year period is reported. These skin diseases are rare (1.3% of our canine case load) and pleomorphic, from localized erythematous and squamous or suppurative lesions (kerions) to extensive or generalized erythematous, alopecic, scaling, crusting and eventually papular and pustular dermatoses. Such dermatophytoses are diagnosed by fungal culture, and in some instances rapidly by dermatohistopathology with periodic acid-Schiff staining (3/3 cases of Microsporum gypseum infection and 5/7 cases of Microsporum persicolor infection). Therapy with topical and/or systemic azole derivatives (topical enilconazole, miconazole, econazole and systemic ketoconazole, respectively) was successful in all cases, with only two relapses, which were easily controlled.     

宁夏肉牛皮肤病原真菌的分离与鉴定

【目的】从患真菌性皮肤病肉牛的毛发、皮屑、痂皮等病料样本中分离皮肤病原真菌并对其进行种属分类鉴定,为牛真菌性皮肤病的诊断及防治提供参考依据。【方法】无菌采集宁夏地区患皮肤病肉牛的毛发、皮屑、痂皮,将病料样本置于载玻片上,滴加10% KOH溶液1滴,静置5 min后进行镜检观察;将病料样本接种于沙氏葡萄糖琼脂培养基（SDA）中进行真菌的分离、纯化培养,对分离的真菌用乳酸酚棉兰染液进行染色观察,根据真菌形态学特征对其进行初步分类;用分离的真菌制备浓度为1×10⁸ cfu·mL^-1的孢子悬液涂抹健康ICR小鼠皮肤,根据其致病性筛选病原真菌;提取病原真菌DNA,对其内部转录间隔区（ITS）进行PCR扩增并测序,登录NCBI GenBank数据库,对病原真菌ITS序列进行同源性比较并构建系统发育树,根据其同源性比较及系统发育分析结果,结合真菌形态学特征对病原真菌作出种属分类鉴定。【结果】病料中含真菌菌丝及链状厚垣孢子,毛发中存在大量小分生孢子;从病料样本中共分离、纯化到23株真菌,分属于毛癣菌属（Trichophyton）、曲霉属（Aspergillus）、横梗霉属（Lichtheimia）、链格孢属（Alternaria）、镰刀菌属（Fusarium）、附球菌属（Epicoccum）、毛霉菌属（Mucor）;小鼠皮肤感染试验表明,分离的毛癣菌属真菌NXGY1、NXGY2具有较强的致病性,试验组小鼠均出现瘙痒、皮屑增多、形成结痂、脱毛等临床症状;毛癣菌属NXGY1、NXGY2 的ITS序列长度大小分别为660、662 bp,其同源性比较及系统发育分析结果表明,NXGY1、NXGY2与疣状毛癣菌(Trichophyton verrucosum)同源性高达99% ,在系统发育树上属于同一分支,遗传距离最近。【结论】根据真菌形态特征及系统发育分析结果,鉴定分离的肉牛皮肤病原真菌NXGY1、NXGY2为疣状毛癣菌(T. verrucosum)。     

致病真菌PCR检测方法的建立

以须癣毛癣菌的基因组DNA为模板,采用真菌通用引物(1TS1、ITS4)PCR扩增rDNA上的ITS段,进行凝胶电泳,结果表明临床样品与相应的培养菌株PCR电泳图谱一致,为准确微量检测致病真菌提供了分子生物学检测方法。     

Antifungal activity evaluation of the constituents of Haliclona baeri and Haliclona cymaeformis, collected from the Gulf of Thailand

A new compound maleimide-5-oxime was isolated, together with 3,4-dihydroxybenzoic acid, tetillapyrone, from the ethyl acetate extract of the marine sponge Haliclona baeri while tetillapyrone, nortetillapyrone, p-hydroxybenzaldehyde and phenylacetic acid were isolated from the ethyl acetate extract of Haliclona cymaeformis, collected from the Gulf of Thailand. The structures of tetillapyrone and nortetillapyrone were re-examined using HMBC correlations. Maleimide-5-oxime, tetillapyrone and nortetillapyrone were found to be inactive against three human tumor cell lines (the estrogen-dependent ER(+) MCF-7, the estrogen-independent ER(−) MDA-MB-231 and NCI-H460. Maleimide-5-oxime, p-hydroxybenzaldehyde, phenylacetic acid, tetillapyrone and nortetillapyrone were evaluated for their growth inhibitory effect against seven yeasts and eight filamentous fungi. Only nortetillapyrone showed antifungal activity, with a preponderance on the dermatophytic filamentous fungi.     

In search of the source of dermatophytosis: Epidemiological analysis of Trichophyton verrucosum infection in llamas and the breeder (case report)

During the last few years, the number of cases of Trichophyton verrucosum isolation from humans suffering from mycoses has been constantly increasing, which is correlated with the presence of an increasing number of outdoor breeding farms. Farmers and their families as well as veterinarians and technicians involved in handling the animals are at a higher risk of infection. One of the most important aims of mycological diagnostics is epidemiological analysis. Typically, the history of the disease is not sufficient to indicate reliably and eliminate the outbreak of infection. PCR fingerprinting methods are a useful tool in this type of analysis, which is presented in this study. The main aim is to present diagnostic and epidemiological analyses of dermatophyte isolates from llamas and their breeder. In two llamas, round alopecia sites or ca. 2‐cm excoriations covered with thickened scaling epidermis were noticed at the border of the head and neck with a distinct tendency towards hair loss. Tinea unguium was noticed in a nail of the breeder's right hand. Direct analysis of the material from the clinical lesions revealed the presence of arthrospores. The macro‐ and micromorphology of the isolates were homogeneous and characteristic for T. verrucosum. The identification analysis based on the ITS sequences confirmed the previous morphological diagnostic examination. The MP‐PCR and MSP‐PCR analysis indicated high invariability of the genomes of the strains isolated from the human and animals. The epidemiological research has indicated an identical source of dermatophyte infection in the breeder and the lamas. To sum up, the number of pets and farm animals is increasing and dermatologists should always be informed about possible dermatophyte transmission sources. The possibility of transmission of zoophilic dermatophytes from humans to animals is a suggestion for further analysis; therefore, this type of transmission should be considered in dermatological studies.     

河北兔场皮肤病原真菌rDNA-ITS序列分析

利用皮肤真菌核糖体内转录间隔区（ITS）序列通用引物,对采自河北省兔场的5种皮肤真菌病进行了PCR扩增,ITS区的克隆、测序、序列变异及遗传进化关系分析。经与GenBank核酸序列数据库数据比对和形态学观察结果表明：有4种真菌被鉴定,分别为须毛癣菌、多聚曲霉、球孢白僵菌和产黄青霉,1种未鉴定;不同病原菌的5.8SrDNA序列高度保守,而ITS区的变异性则较高。该研究确定ITS区序列分析可用于兔皮肤病原真菌的分离。     

Evaluation of the dermatophyte test medium RapidVet-D

The performance of the dermatophyte test medium (DTM) RapidVet-D was assessed using hair samples collected from experimentally infected guinea pigs. Three dermatophyte species were included in the study: Microsporum canis , Trichophyton mentagrophytes and Trichophyton equinum . DTM substrates were inoculated with infected hairs and scales, incubated at 18, 21, 24, 27 or 37 °C and examined daily for 15 days. The rapidity of colour change was clearly related to the incubation temperature and to the number of infected hairs deposited on the reactive substrates. With the optimum incubation temperature 27 °C, a systematic colour change could be observed only a few days post-inoculation: 3 days with M. canis infected hairs, 4 days with T. equinum and 5 days with T. mentagrophytes .     

Evaluation of topically applied enilconazole for the treatment of dermatophytosis in a Persian cattery

Many Persian catteries have long-standing dermatophyte infections and are particularly difficult to treat. Enilconazole is a topical antifungal agent that has demonstrated good efficacy in recent studies. Twenty-two Persian cats naturally infected with Microsporum canis in a breeding cattery were treated with topical 0.2% enilconazole and monitored for 180 days. The treatments were repeated every 3 days for a total of eight applications. All the cats improved clinically and became culture negative by day 28. By day 180, four cats had developed clinical dermatophytosis and all cats had positive fungal cultures. In this study, topical 0.2% enilconazole was generally well tolerated but may have caused hypersalivation, idiopathic muscle weakness and slightly elevated serum alanine aminotransferase (ALT) concentrations. This study suggests that enilconazole may be used safely with little risk to the young, aged and gravid animals.