Bouchali Rayan, Marjolet Laurence, Mondamert Leslie, Chonova Teofana, Ribun Sébastien, Laurent Elodie, Bouchez Agnès, Labanowski Jérôme, Cournoyer Benoit
UMR Ecologie Microbienne, CNRS 5557, INRAE 1418, Research Group «Bacterial Opportunistic Pathogens and Environment», VetAgro Sup, Aisle 3, 1st Floor, 69280 Marcy L'Etoile, France.
Institut de Chimie des Milieux et des Matériaux de Poitiers (IC2MP), École Nationale Supérieure d'Ingénieurs (ENSIP), UMR CNRS 7285, Université de Poitiers, 86000 Poitiers, France.
Microorganisms. 2023 Apr 2;11(4):922. doi: 10.3390/microorganisms11040922.
The ability of WWTP outflow bacteria at colonizing rock surfaces and contributing to the formation of river epilithic biofilms was investigated. Bacterial community structures of biofilms (b-) developing on rocks exposed to treated wastewaters (TWW) of a hospital (HTWW) and a domestic (DTWW) clarifier, and to surface waters of the stream located at 10 m, 500 m, and 8 km from the WWTP outlet, were compared. Biofilm bacterial contents were analyzed by cultural approaches and a -based DNA metabarcoding analytical scheme. Co-occurrence distribution pattern analyses between bacterial datasets and eighteen monitored pharmaceuticals were performed. Higher concentrations of iohexol, ranitidine, levofloxacin, and roxithromycin were observed in the b-HTWW while atenolol, diclofenac, propranolol, and trimethoprim were higher in the b-DTWW. MPN growth assays showed recurrent occurrences of and among these biofilms. An enrichment of multi-resistant cells was observed in the hospital sewer line. MPN values were negatively correlated to roxithromycin concentrations. The DNA metabarcoding analyses confirmed these trends and allowed an additional tracking of more than 90 species from 24 genera. Among the recorded 3082 ASV (amplicon sequence variants), 41% were allocated to the . Significant differences through ANOSIM and DESeq2 statistical tests were observed between ASV recovered from b-HTWW, b-DTWW, and epilithic river biofilms. More than 500 ASV were found restricted to a single sewer line such as those allocated to and being strictly found in the b-HTWW file. Several significant correlations between ASV counts per species and pharmaceutical concentrations in biofilms were recorded such as those of being positively correlated with trimethoprim concentrations. A source tracking analysis showed the b-DTWW and b-HTWW ASV to have contributed, respectively, at up to 35% and 2.5% of the epilithic river biofilm -taxa recovered downstream from the WWTP outlet. Higher contributions of TWW taxa among epilithic biofilms were recorded closer to the WWTP outlet. These analyses demonstrated a coalescence of WWTP sewer communities with river freshwater taxa among epilithic biofilms developing downstream of a WWTP outlet.
研究了污水处理厂流出物中的细菌在岩石表面定殖并促进河流附石生物膜形成的能力。比较了在暴露于医院（HTWW）和生活污水处理厂（DTWW）澄清池的处理后废水（TWW）以及距离污水处理厂出水口10米、500米和8公里处溪流地表水的岩石上形成的生物膜（b-）的细菌群落结构。通过培养方法和基于16S rRNA基因的DNA宏条形码分析方案分析生物膜细菌含量。对细菌数据集与18种监测药物之间的共现分布模式进行了分析。在b-HTWW中观察到碘海醇、雷尼替丁、左氧氟沙星和罗红霉素的浓度较高，而在b-DTWW中阿替洛尔、双氯芬酸、普萘洛尔和甲氧苄啶的浓度较高。MPN生长试验表明这些生物膜中反复出现粪肠球菌和屎肠球菌。在医院污水管道中观察到多重耐药肠球菌细胞的富集。MPN值与罗红霉素浓度呈负相关。16S rRNA基因DNA宏条形码分析证实了这些趋势，并允许额外追踪来自24个属的90多种物种。在记录的3082个ASV（扩增子序列变体）中，41%被归类为变形菌门。通过ANOSIM和DESeq2统计检验观察到从b-HTWW、b-DTWW和河流附石生物膜中回收的ASV之间存在显著差异。发现500多个ASV仅限于单一污水管道，例如那些严格在b-HTWW文件中发现的被归类为假单胞菌属和不动杆菌属的ASV。记录了生物膜中每个物种的16S rRNA基因ASV计数与药物浓度之间的几个显著相关性，例如粪肠球菌与甲氧苄啶浓度呈正相关。一项来源追踪分析表明，b-DTWW和b-HTWW的16S rRNA基因ASV分别对污水处理厂出水口下游回收的河流附石生物膜细菌分类群贡献高达35%和2.5%。在更靠近污水处理厂出水口的附石生物膜中记录到污水处理厂分类群的贡献更高。这些分析表明了在污水处理厂出水口下游形成的附石生物膜中，污水处理厂污水群落与河流淡水分类群的融合。
