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虫生真菌节丛孢的分离与鉴定。

Isolation and characterization of the nematophagous fungus Arthrobotrys conoides.

机构信息

Department of Veterinary Medicine, Universidade Estadual do Centro Oeste, UNICENTRO, Rua Simeão Camargo Varela de Sá, 03. CEDETEG, Guarapuava, PR, CEP 85040-080, Brazil.

出版信息

Parasitol Res. 2013 Jan;112(1):177-85. doi: 10.1007/s00436-012-3123-3. Epub 2012 Sep 20.

Abstract

The spread of organic farming and the development of resistance to anthelmintics by parasites, especially in small ruminants, have necessitated the search for alternative methods of nematode control. Biological control using nematophagous fungi is one option; however, few studies have been conducted with native strains. The present study was divided into two phases. In the first phase, we aimed to isolate, identify, and assess the in vitro predatory activity of nematophagous fungi that had been isolated on Trichostrongylidae third-instar larvae. In the second phase, the isolate with superior predatory activity in vitro was molecularly characterized, and its morphological plasticity was observed using scanning electron microscopy (SEM) on Haemonchus third-instar larvae. Of the 56 soil samples from different regions of Paraná State, Brazil, 57 fungal strains were recovered, of which four exhibited predatory activity. Two pure isolates were obtained: the CED and LIN strains. After demonstrating 96.35 % predatory activity for the CED strain, this strain was selected and characterized using molecular criteria by sequencing the rDNA internal transcribed spacer and was identified as Arthrobotrys conoides (GenBank ID: JN191309). Morphological patterns in this strain during the interaction between the fungus and the nematode were revealed by SEM, in which two extensions of the infection bulb that was used to pierce the nematode's cuticle were clearly visible.

摘要

有机农业的推广和寄生虫(尤其是小反刍动物)对驱虫剂的耐药性发展,使得人们有必要寻找替代线虫控制方法。利用食线虫真菌进行生物防治是一种选择;然而,针对本地菌株的研究很少。本研究分为两个阶段。在第一阶段,我们旨在分离、鉴定并评估已从捻转血矛线虫 3 龄幼虫上分离出的食线虫真菌的体外捕食活性。在第二阶段,对具有优越体外捕食活性的分离株进行分子特征描述,并使用扫描电子显微镜 (SEM) 观察其在绵羊仰口线虫 3 龄幼虫上的形态可塑性。在巴西巴拉那州不同地区的 56 个土壤样本中,共回收了 57 株真菌菌株,其中 4 株表现出捕食活性。获得了两个纯培养物:CED 和 LIN 菌株。在 CED 菌株表现出 96.35%的捕食活性后,选择该菌株并通过测序 rDNA 内转录间隔区进行分子标准特征描述,鉴定为节丛孢属(Arthrobotrys conoides)(GenBank ID:JN191309)。通过 SEM 揭示了该菌株与线虫相互作用过程中的形态模式,其中可以清楚地看到用于刺穿线虫角质层的感染球的两个延伸。

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