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曲霉病,家禽的一种自然感染:烟曲霉分离株的真菌学和分子特征及胶霉毒素的测定

Aspergillosis, a Natural Infection in Poultry: Mycological and Molecular Characterization and Determination of Gliotoxin in Aspergillus fumigatus Isolates.

作者信息

de Oca Verónica Montes, Valdés Sara Esther, Segundo Carolina, Gómez Gabriela Guadalupe, Ramírez José, Cervantes Roberto Arnulfo

机构信息

A Microbiology and Immunology Department, Faculty of Veterinary Medicine and Animal Science, National Autonomous University of Mexico, Avenida Universidad 3000, Ciudad Universitaria 04510, Mexico City, Mexico.

B Laboratory of Quality Food Technology, Faculty of Higher Studies, Campus Cuautitlán, National Autonomous University of Mexico, Avenida 1o de Mayo S/N, Santa María las Torres, Campo 1 54740 Cuautitlán Izcalli, Mexico.

出版信息

Avian Dis. 2017 Mar;61(1):77-82. doi: 10.1637/11496-092016-Reg.

Abstract

Aspergillosis affects all types of birds; it causes the loss of specimens with high ecologic value and also leads to significant economic losses within the poultry industry. The main etiologic agent is Aspergillus fumigatus , a filamentary fungus with multiple virulence factors, such as gliotoxin (GT), which is an immunosuppressive epipolythiodioxopiperazine molecule. Necropsy was performed on 73 poultry from different provenances, all of which presented with a respiratory semiology compatible with aspergillosis. A mycological culture was performed on the injured lungs of diseased birds, as was chloroform extraction of the GT, a thin-layer chromatography analysis (TLC), and a histopathology analysis with hematoxylin-eosin and Grocott stainings. The A. fumigatus identification was confirmed by PCR, where the ITS 1 5.1-5.8S-ITS 2 fragment of the rDNA complex was amplified. The in vitro GT production was studied by TLC in the recovered isolates from A. fumigatus . Seven isolates of A. fumigatus were obtained and in six of them, GT-like compounds were detected. In a lung sample, a compound with the same retention time (RF) as the reference GT was detected; whereas RF compounds different from the GT standard were observed in three lung samples.

摘要

曲霉菌病会影响所有鸟类;它导致具有高生态价值的鸟类标本损失,还会给家禽业造成重大经济损失。主要病原体是烟曲霉,这是一种具有多种毒力因子的丝状真菌,比如胶霉毒素(GT),它是一种免疫抑制性的表聚硫代二氧哌嗪分子。对73只来自不同产地的家禽进行了尸检,所有家禽均表现出与曲霉菌病相符的呼吸症状。对患病鸟类的受损肺部进行了真菌培养,以及GT的氯仿萃取、薄层色谱分析(TLC),还有苏木精-伊红染色和Grocott染色的组织病理学分析。通过PCR确认了烟曲霉的鉴定结果,其中rDNA复合体的ITS 1 5.1-5.8S-ITS 2片段被扩增。通过TLC对从烟曲霉中分离得到的菌株进行体外GT产量研究。获得了7株烟曲霉菌株,其中6株检测到了GT样化合物。在一个肺样本中,检测到一种与参考GT具有相同保留时间(RF)的化合物;而在三个肺样本中观察到了与GT标准品不同RF的化合物。

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