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分析寄生虫真菌的安全性:对其毒力特征及其与禽类肠道微生物群落相互作用的初步了解。

Analyzing the safety of the parasiticide fungus : first insights on its virulence profile and interactions with the avian gut microbial community.

机构信息

CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.

Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal.

出版信息

Microbiol Spectr. 2024 May 2;12(5):e0407823. doi: 10.1128/spectrum.04078-23. Epub 2024 Mar 27.

Abstract

Parasiticide fungi are considered an accurate, sustainable, and safe solution for the biocontrol of animal gastrointestinal (GI) parasites. This research provides an initial characterization of the virulence of the native parasiticide fungus (FMV-FR1) and an assessment of its impact on birds' gut microbes. The genome of this fungus was sequenced to identify the genes coding for virulence factors. Also, this fungus was checked for the phenotypic expression of proteinase, lecithinase, DNase, gelatinase, hemolysin, and biofilm production. Finally, an trial was developed based on feeding spores to laying hens and peacocks three times a week. Bird feces were collected for 3 months, with total genomic DNA being extracted and subjected to long-read 16S and 25S-28S sequencing. Genes coding for an iron permease (FTR1), iron receptors (FOB1 and FOB2), ADP-ribosylation factors (ARFs) (ARF2 and ARF6), and a GTPase (CDC42) were identified in this genome. Also, this fungus was positive only for lecithinase activity. The field trial revealed a fecal microbiome dominated by Firmicutes and Proteobacteria in laying hens, and Firmicutes and Bacteroidetes in peacocks, whereas the fecal mycobiome of both bird species was mainly composed of Ascomycetes and Basidiomycetes fungi. Bacterial and fungal alpha-diversities did not differ between sampling time points after administrations ( = 0.62 and = 0.15, respectively). Although findings from this research suggest the lack of virulence of this parasiticide isolate, more complementary and research is needed to conclude about the safety of its administration to birds, aiming at controlling their GI parasites.IMPORTANCEA previous study revealed that the native isolate (FMV-FR1) can develop parasiticide activity toward coccidia oocysts, one of the most pathogenic GI parasites in birds. However, ensuring its safety for birds is of utmost importance, namely by studying its virulence profile and potential effect on commensal gut microbes. This initial study revealed that although this isolate had genes coding for four types of virulence factors-iron permease, iron receptors, ADP-ribosylation factors, and GTPase-and only expressed phenotypically the enzyme lecithinase, the administration of its spores to laying hens and peacocks did not interfere with the abundances and diversities of their gut commensal bacteria and fungi. Although overall results suggest the lack of virulence of this isolate, more complementary research is needed to conclude about the safety of its administration to birds in the scope of parasite biocontrol programs.

摘要

寄生虫真菌被认为是一种针对动物胃肠道 (GI) 寄生虫的生物防治的准确、可持续和安全的解决方案。本研究初步描述了本地寄生虫真菌 (FMV-FR1) 的毒力,并评估了其对鸟类肠道微生物的影响。该真菌的基因组已被测序,以鉴定编码毒力因子的基因。此外,还检查了该真菌蛋白酶、卵磷脂酶、DNase、明胶酶、溶血素和生物膜产生的表型表达。最后,基于每周给产蛋母鸡和孔雀喂食孢子三次,开展了一项试验。收集了 3 个月的鸟类粪便,提取总基因组 DNA 并进行长读 16S 和 25S-28S 测序。在这个基因组中鉴定出铁渗透酶 (FTR1)、铁受体 (FOB1 和 FOB2)、ADP-核糖基化因子 (ARFs) (ARF2 和 ARF6) 和 GTPase (CDC42) 的编码基因。此外,该真菌仅对卵磷脂酶活性呈阳性。田间试验表明,产蛋母鸡的粪便微生物群主要由厚壁菌门和变形菌门组成,而孔雀的粪便微生物群主要由厚壁菌门和拟杆菌门组成,而这两种鸟类的粪便真菌群主要由子囊菌门和担子菌门真菌组成。细菌和真菌的α多样性在给药后不同时间点没有差异(=0.62 和=0.15)。尽管这项研究的结果表明这种寄生虫分离株没有毒力,但需要更多的互补和研究来得出关于将其施用于鸟类以控制其 GI 寄生虫的安全性的结论。

重要性

先前的研究表明,本地分离株(FMV-FR1)可以对球虫卵囊产生寄生虫活性,球虫卵囊是鸟类中最具致病性的 GI 寄生虫之一。然而,确保其对鸟类的安全性至关重要,即通过研究其毒力谱及其对共生肠道微生物的潜在影响。这项初步研究表明,尽管该分离株具有编码四种类型毒力因子(铁渗透酶、铁受体、ADP-核糖基化因子和 GTPase)的基因,并且仅表型上表达了卵磷脂酶,但向产蛋母鸡和孔雀投喂其孢子不会干扰其肠道共生细菌和真菌的丰度和多样性。尽管总体结果表明该分离株缺乏毒力,但需要更多的互补研究来得出关于将其施用于鸟类以控制其 GI 寄生虫的生物防治计划中安全性的结论。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb9/11064519/bbe804ae9de7/spectrum.04078-23.f001.jpg

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