<br>使用 18S rRNA 和鸟枪法宏基因组学分析废水中的致病原虫及其功能途径,Science of the Total Environment

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使用 18S rRNA 和鸟枪法宏基因组学分析废水中的致病原虫及其功能途径
Science of the Total Environment ( IF 8.2 ) Pub Date : 2023-12-26 , DOI: 10.1016/j.scitotenv.2023.169602
Nonsikelelo P Mthethwa-Hlongwa ₁ , Isaac D Amoah ₂ , Andres Gomez ₃ , Sam Davison ₃ , Poovendhree Reddy ₄ , Faizal Bux ₅ , Sheena Kumari ₅

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尽管进行了广泛的研究，但人们对环境样本中真核原生生物的组成知之甚少。这是由于寄生虫浓度低、寄生虫多样性复杂以及缺乏合适的参考数据库和标准化方案。为了弥补这一知识差距，本研究使用 18S rRNA 短扩增子和鸟枪法宏基因组测序方法来分析原生动物微生物群落及其在从南非不同地区收集的经过处理和未经处理的废水样本中的功能途径。结果表明，原生动物多样性（香农指数 P 值 = 0.03）和分类组成（PERMANOVA，P 值 = 0.02）主要由废水样品类型（已处理和未处理）和地理位置驱动。然而，这些污水处理厂还被发现含有原生动物寄生虫的核心群落。未经处理的废水样本显示，主要存在于人类和动物中常见的自由生活、寄生和潜在致病性原生生物，范围从 Alveolata (27%) 门（Apicomplexa 和 Ciliophora）到 Excavata (3.88%)（Discoba 和 Parasalia）和变形虫 (2.84%)（内阿米巴和棘阿米巴）。对未经处理的废水样本的子集进行鸟枪法宏基因组学分析，证实了对公共卫生具有重要意义的原生动物的存在，包括隐孢子虫 (3.48%)、囊状内阿米巴 (6.58%)、人芽囊原虫 (2.91%)、格鲁伯氏耐格里阿米巴 (2.37%)、弓形虫 (1.98 %)、环孢子虫 (1.30 %) 和贾第鞭毛虫 (0.31 %)。鉴定出与致病原生动物相关的毒力基因家族，例如丝氨酸/苏氨酸蛋白磷酸酶和粘蛋白脱硫硫酸酯酶。此外，丰富的途径包括硫胺素二磷酸生物合成 III、血红素生物合成、甲基赤藓糖醇 4-磷酸途径、甲基赤藓糖醇磷酸 (MEP) 和戊糖磷酸途径。这些发现表明，原生动物病原体可能在污水处理厂内具有代谢和生长潜力，如果在释放前消毒不充分，将构成传播给人类和动物的严重风险。这项研究为未来对废水中多种原生动物群落的调查提供了基线，这对公共卫生具有重要意义。

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Profiling pathogenic protozoan and their functional pathways in wastewater using 18S rRNA and shotgun metagenomics

Despite extensive research, little is known about the composition of eukaryotic protists in environmental samples. This is due to low parasite concentrations, the complexity of parasite diversity, and a lack of suitable reference databases and standardized protocols. To bridge this knowledge gap, this study used 18S rRNA short amplicon and shotgun metagenomic sequencing approaches to profile protozoan microbial communities as well as their functional pathways in treated and untreated wastewater samples collected from different regions of South Africa. Results demonstrated that protozoan diversity (Shannon index P-value = 0.03) and taxonomic composition (PERMANOVA, P-value = 0.02) was mainly driven by the type of wastewater samples (treated & untreated) and geographic location. However, these WWTPs were also found to contain a core community of protozoan parasites. The untreated wastewater samples revealed a predominant presence of free-living, parasitic, and potentially pathogenic protists typically found in humans and animals, ranging from Alveolata (27 %) phylum (Apicomplexa and Ciliophora) to Excavata (3.88 %) (Discoba and Parasalia) and Amoebozoa (2.84 %) (Entamoeba and Acanthamoeba). Shotgun metagenomics analyses in a subset of the untreated wastewater samples confirmed the presence of public health-importance protozoa, including Cryptosporidium species (3.48 %), Entamoeba hystolitica (6.58 %), Blastocystis hominis (2.91 %), Naegleria gruberi (2.37 %), Toxoplasma gondii (1.98 %), Cyclospora cayetanensis (1.30 %), and Giardia intestinalis (0.31 %). Virulent gene families linked to pathogenic protozoa, such as serine/threonine protein phosphatase and mucin-desulfating sulfatase were identified. Additionally, enriched pathways included thiamine diphosphate biosynthesis III, heme biosynthesis, Methylerythritol 4-Phosphate Pathway, methyl erythritol phosphate (MEP), and pentose phosphate pathways. These findings suggest that protozoan pathogens may possess metabolic and growth potential within WWTPs, posing a severe risk of transmission to humans and animals if inadequately disinfected before release. This study provides a baseline for the future investigation of diverse protozoal communities in wastewater, which are of public health importance.

更新日期：2023-12-26

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