天山北坡绵羊面部湿疹高发草场病原真菌分离及PCR-DGGE分析

doi:10.11843/j.issn.0366-6964.2014.10.019

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (10): 1718-1725.doi: 10.11843/j.issn.0366-6964.2014.10.019

天山北坡绵羊面部湿疹高发草场病原真菌分离及PCR-DGGE分析

刘良波，张豫，孙志华，欧科鹏，孙焕林，剡根强^*

（石河子大学动物科技学院，石河子 832003）

收稿日期:2014-02-07 出版日期:2014-10-23 发布日期:2014-10-23
通讯作者: 剡根强，教授，E-mail：ygq58@shzu.edu.cn
作者简介:刘良波(1980-)，男，四川大竹人，助教，博士生，主要从事遗传抗病机理研究，E-mail：13768963@qq.com，Tel：0993-2058583
基金资助:
国家科技支撑计划课题（2012BAD43B02）

Isolation and PCR-DGGE Studies of the Fungal Pathogens from Pastures with High Incidence of Sheep Facial Eczema on the Northern Slope of Tianshan Mountains

LIU Liang-bo，ZHANG Yu，SUN Zhi-hua，OU Ke-peng，SUN Huan-lin，YAN Gen-qiang^*

（College of Animal Science & Technology，Shihezi University，Shihezi 832003，China）

Received:2014-02-07 Online:2014-10-23 Published:2014-10-23

摘要/Abstract

摘要：

旨在探讨天山北坡放牧绵羊面部湿疹发病区草场真菌的种类及多样性，查明疾病发生与放牧地理位置、真菌群落分布的相关性。调查发病地区的地理环境特征，采集发病季节绵羊面部湿疹发病区域和非发病区域牧草和土壤样本，进行真菌分离培养，形态学观察，真菌种类及分布统计分析；提取真菌总DNA，利用18S rDNA的变性梯度凝胶电泳（DGGE）和测序技术，进行真菌多样性分析。结果显示发病地区地理位置为N44.016～44.022，E85.792～85.800，海拔925～1 039 m，是山前倾斜洪积扇平原，属典型的温带干旱大陆性气候，是当地重要的春秋草场。真菌分离培养、PCR-DGGE及测序分析结果显示该地区真菌类群主要包括10种真菌，其中链格孢菌属、纸皮司霉属、镰刀菌属、曲霉属、青霉属在发病区域分离率较高，纸皮司霉属真菌分离率最高，与发病绵羊消化道分离菌相一致。试验结果表明该地区引起绵羊肝原性光过敏面部湿疹的真菌可能为链格孢菌属、纸皮司霉属、镰刀菌属、曲霉属真菌；发病区放牧的地理位置、真菌分布与疫病发生具有相关性，为进一步揭示放牧绵羊面部湿疹的发病原因及影响因素提供了依据。

Abstract:

This paper was undertaken to investigate fungi species which cause facial eczema (FE) of grazing sheep from pastures with high incidence of FE on the northern slope of Tianshan Mountains，and analyze their diversity，then identify the relationship among disease occurrence grazing location，and community distribution of fungal correlation.After investigating the features of geographical environment in the FE incidence area，forage and soil samples were cultured from incidence areas and non epidemic areas during FE season.Fungi were isolated and cultured for further studies，such as morphological observation，species and distribution analysis.Total fungal DNAs were extracted，and then PCR，denaturing gradient gel electrophoresis (DGGE) and DNA sequencing of 18S rDNA were used to explore fungal diversity.The results showed that the FE incidence area located at inclined plains of piedmont alluvial fan，N44.016-44.022，E85.792-85.800，and altitude of 925-1 039 m，where expressing typical temperate continental arid climate.They are important local spring- and autumn pastures.Fungal cultivation，PCR-DGGE and sequencing analysis of 18S rDNA showed that there found 10 kinds of fungi in above areas.Separation rates of Alternaria，Pithomyces chartarum，Fusarium，Aspergillus，Penicillium were higher，and that of Pithomyces chartarum was the highest，these results were identical with the isolated fungi in digestive tract of FE sheep.Our research indicated that Alternaria spp.，pithomyces chartarum spp., Fusarium and aspergillus maybe the responsible pathogens of hepatogenic light allergic FE.There were the correlation among grazing location，fungi distribution and FE incidence，which provides a basis for revealing the pathogenesis and the influence factors of FE in grazing sheep.

中图分类号:

S858.265.4

刘良波，张豫，孙志华，欧科鹏，孙焕林，剡根强. 天山北坡绵羊面部湿疹高发草场病原真菌分离及PCR-DGGE分析[J]. 畜牧兽医学报, 2014, 45(10): 1718-1725.

LIU Liang-bo，ZHANG Yu，SUN Zhi-hua，OU Ke-peng，SUN Huan-lin，YAN Gen-qiang. Isolation and PCR-DGGE Studies of the Fungal Pathogens from Pastures with High Incidence of Sheep Facial Eczema on the Northern Slope of Tianshan Mountains[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(10): 1718-1725.

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天山北坡绵羊面部湿疹高发草场病原真菌分离及PCR-DGGE分析

Isolation and PCR-DGGE Studies of the Fungal Pathogens from Pastures with High Incidence of Sheep Facial Eczema on the Northern Slope of Tianshan Mountains

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