Wastewater treatment alters microbial colonization of microplastics

Abstract

Microplastics are ubiquitous contaminants in aquatic habitats globally, and wastewater treatment plants (WWTPs) are point sources of microplastics. Within aquatic habitats microplastics are colonized by microbial biofilms, which can include pathogenic taxa and taxa associated with plastic breakdown. Microplastics enter WWTPs in sewage and exit in sludge or effluent, but the role that WWTPs play in establishing or modifying microplastic bacterial assemblages is unknown. We analyzed microplastics and associated biofilms in raw sewage, effluent water, and sludge from two WWTPs. Both plants retained >99% of influent microplastics in sludge, and sludge microplastics showed higher bacterial species richness and higher abundance of taxa associated with bioflocculation (e.g. Xanthomonas) than influent microplastics, suggesting that colonization of microplastics within the WWTP may play a role in retention. Microplastics in WWTP effluent included significantly lower abundances of some potentially pathogenic bacterial taxa (e.g. Campylobacteraceae) compared to influent microplastics; however, other potentially pathogenic taxa (e.g. Acinetobacter) remained abundant on effluent microplastics, and several taxa linked to plastic breakdown (e.g. Klebsiella, Pseudomonas, and Sphingomonas) were significantly more abundant on effluent compared to influent microplastics. These results indicate that diverse bacterial assemblages colonize microplastics within sewage and that WWTPs can play a significant role in modifying the microplastic-associated assemblages, which may affect the fate of microplastics within the WWTPs and the environment.

Fig 1. Relative abundance of microplastic particle types within each sample type (sewage, effluent, and sludge) from each of two WWTPs (Bartlett and Woodridge).

Each data point represents the mean value (n = 4).

Fig 2. Number of operational taxonomic units (based on 97% identity in 16S rRNA amplicons) observed within bacterial communities attached to microplastic particles collected from three sample types (sewage, effluent, and sludge) from two WWTPs (Bartlett and Woodridge).

Each data point represents the mean value (n = 4) with error bars representing standard error.

Fig 3. nMDS ordination of bacterial assemblage composition for biofilms attached to microplastic particles collected from three sample types (sewage, effluent, and sludge) from two WWTPs (Bartlett and Woodridge).

Each point represents the bacterial assemblage from one individual sample. Bacterial assemblage analysis was based on high-throughput amplicon sequencing of partial 16 rRNA genes, clustering sequences into OTUs (97% sequence identity) and comparison of assemblages based on the theta index. Stress value of ordination = 0.2997.

Fig 4. Average relative abundance of the 25 most abundant bacterial families within microplastic-attached bacterial assemblages from three sample types (sewage, effluent, and sludge).

Each bar represents the mean (n = 8). Bacterial families were identified based on high-throughput amplicon sequencing of partial 16 rRNA genes.