Wastewater treatment plants (WWTPs) receive wastewater from various sources. Despite wastewater treatment aiming to remove contaminants, microplastics persist. Plastic surfaces are quickly colonized by microbial biofilm ("plastispheres"). Plastisphere communities are suggested to promote the spread and survival of potential human pathogens, suggesting that the transfer of plastispheres from wastewater to the environment could pose a risk to human and environmental health. The study aimed to identify pathogens in wastewater plastispheres, specifically food-borne pathogens, in addition to characterizing the taxonomic diversity and composition of the wastewater plastispheres. Plastispheres that accumulated on polypropylene (PP), polyvinyl chloride (PVC), and high-density polyethylene propylene (HDPE) surfaces exposed to raw and treated wastewater were analyzed via cultivation methods, quantitative reverse transcription PCR (RT‒qPCR) and 16S rRNA amplicon sequencing. RT‒qPCR revealed the presence of potential foodborne pathogenic bacteria and viruses, such as Listeria monocytogenes, Escherichia coli, norovirus, and adenovirus. Viable isolates of the emerging pathogenic species Klebsiella pneumoniae and Acinetobacter spp. were identified in the plastispheres from raw and treated wastewater, indicating that potential pathogenic bacteria might survive in the plastispheres during the wastewater treatment. These findings underscore the potential of plastispheres to harbor and disseminate pathogenic species, posing challenges to water reuse initiatives. The taxonomic diversity and composition of the plastispheres, as explored through 16S rRNA amplicon sequencing, were significantly influenced by the wastewater environment and the duration of time the plastic spent in the wastewater. In contrast, the specific plastic material did not influence the bacterial composition, while the bacterial diversity was affected. Without efficient wastewater treatment and proper plastic waste management, wastewater could act as a source of transferring plastic-associated pathogens into the food chain and possibly pose a threat to human health. Continued research and innovation are essential to improve the removal of microplastics and associated pathogenic microorganisms in wastewater.
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