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The frequency of plastic debris entering agricultural land is likely going to increase due to increased discharge into surface waters and more frequent flood events. Microbial biofilm on the surfaces of plastic pollution (known as the 'plastisphere') in freshwater environments often includes human pathogenic bacteria capable of causing disease. Pathogens have been detected on the surface of plastics in freshwater environments, but it is yet to be determined whether plastic debris can also transport pathogens into agricultural fields during flooding. Therefore, this study quantified the presence of viable pathogenic bacteria on the surface of plastic pollution at five agricultural fields along two rivers. All visible plastic debris, including sewage-associated plastic waste, were collected along a perpendicular 100 m transect from the riparian zone into each field. All plastic pieces were screened for five target bacteria (Escherichia coli, intestinal enterococci, Salmonella spp., Campylobacter spp., and Klebsiella spp.) using selective media, and positively identified colonies subsequently tested for antimicrobial resistance. In all five fields, there were higher volumes of plastic in the areas closer to the river, with 75% ± 24% of plastic collected within 30 m from the riverbank. Overall, 49% of all plastic collected in agricultural fields was colonised by phenotypically positive colonies for at least one or more target bacteria, with resistance to commonly prescribed antibiotics detected among several of these target bacteria. Therefore, the transport of contaminated plastic debris from fluvial floodwater into agricultural fields could pose an as yet unquantified risk of introducing potentially harmful bacteria into agricultural systems and the ultimately into the food chain., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Plastic waste is found with increasing frequency in the environment, in low- and middle-income countries. Plastic pollution has increased concurrently with both economic development and rapid urbanisation, amplifying the effects of inadequate waste management. Distinct microbial communities can quickly colonise plastic surfaces in what is collectively known as the 'plastisphere'. The plastisphere can act as a reservoir for human pathogenic bacteria, including Salmonella enterica sp. (such as S. Typhimurium), which can persist for long periods, retain pathogenicity, and pose an increased public health risk. Through employing a novel mesocosm setup, we have shown here that the plastisphere provides enhanced protection against environmental pressures such as ultraviolet (UV) radiation and allows S. Typhimurium to persist at concentrations (>1 × 10 3  CFU/ml) capable of causing human infection, for up to 28 days. Additionally, using a Galleria Mellonella model of infection, S. Typhimurium exhibits greater pathogenicity following recovery from the UV-exposed plastisphere, suggesting that the plastisphere may select for more virulent variants. This study demonstrates the protection afforded by the plastisphere and provides further evidence of environmental plastic waste acting as a reservoir for dangerous clinical pathogens. Quantifying the role of plastic pollution in facilitating the survival, persistence, and dissemination of human pathogens is critical for a more holistic understanding of the potential public health risks associated with plastic waste., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Plastic waste is ubiquitous in the environment and there are increasing reports of such waste being colonised by human pathogens. However, the ability of pathogens to persist on plastics for long periods, and the risk that they pose to human health, is unknown. Here, under simulated environmental conditions, we aimed to determine if pathogenic bacteria can retain their virulence following a prolonged period on plastic. Using antibiotic selection and luciferase expression for quantification, we show that clinically important strains of E. coli can survive on plastic for at least 28-days. Importantly, these pathogens also retained their virulence (determined by using a Galleria mellonella model as a surrogate for human infection) and in some cases, had enhanced virulence following their recovery from the plastisphere. This indicates that plastics in the environment can act as reservoirs for human pathogens and could facilitate their persistence for extended periods of time. Most importantly human pathogens in the plastisphere are capable of retaining their pathogenicity. Pathogens colonising environmental plastic waste therefore pose a heightened public health risk, particularly in areas where people are exposed to pollution., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Large quantities of microplastics are regularly discharged from wastewater treatment plants (WWTPs) into the aquatic environment. Once released, these plastics can rapidly become colonised by microbial biofilm, forming distinct plastisphere communities which may include potential pathogens. We hypothesised that the protective environment afforded by the plastisphere would facilitate the survival of potential pathogens during transitions between downstream environmental matrices and thus increase persistence and the potential for environmental dissemination of pathogens. The survival of Escherichia coli, Enterococcus faecalis and Pseudomonas aeruginosa colonising polyethylene or glass particles has been quantified in mesocosm incubation experiments designed to simulate, (1) the direct release of microplastics from WWTPs into freshwater and seawater environments; and (2) the movement of microplastics downstream following discharge from the WWTP through the river-estuary-marine-beach continuum. Culturable E. coli, E. faecalis and P. aeruginosa were successfully able to survive and persist on particles whether they remained in one environmental matrix or transitioned between different environmental matrices. All three bacteria were still detectable on both microplastic and glass particles after 25 days, with higher concentrations on microplastic compared to glass particles; however, there were no differences in bacterial die-off rates between the two materials. This potential for environmental survival of pathogens in the plastisphere could facilitate their transition into places where human exposure is greater (e.g., bathing waters and beach environments). Therefore, risks associated with pathogen-microplastic co-pollutants in the environment, emphasises the urgency for updated regulations on wastewater discharge and the management of microplastic generation and release., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Microplastics in wastewater and surface water rapidly become colonised by microbial biofilm. Such 'plastisphere' communities are hypothesised to persist longer and be disseminated further in the environment and may act as a vector for human pathogens, particularly as microplastics entering wastewater treatment plants are exposed to high concentrations of pathogenic bacteria. However, the potential for human viral pathogens to become associated with the plastisphere has never before been quantified. Here, we have used rotavirus (RV) SA11 (a non-enveloped enteric virus) and the enveloped bacteriophage Phi6 as model viruses to quantify binding and recovery from biofilm-colonised microplastic pellets in three different water treatments (filtered and non-filtered surface water, and surface water with added nutrients). Viruses associated with biofilm-colonised pellets were more stable compared to those remaining in the water. While infectious particles and genome copies of RV remained stable over the 48 h sampling period, Phi6 stability was highly impacted, with a reduction ranging from 2.18 to 3.94 log 10 . Virus particles were protected against inactivation factors when associated with the biofilm on microplastic surfaces, and when there was a high concentration of particulate matter in the liquid phase. Although our results suggest that the presence of an envelope may limit virus interaction with the plastisphere, the ability to recover both enveloped and non-enveloped infectious viruses from colonised microplastic pellets highlights an additional potential public health risk of surface waters becoming contaminated with microplastics, and subsequent human exposure to microplastics in the environment., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)