Microplastic biodegradability dependent responses of plastisphere antibiotic resistance to simulated freshwater-seawater shift in onshore marine aquaculture zones.

MPs carrying ARGs can travel between freshwater and seawater due to intensive land-sea interaction in onshore marine aquaculture zones (OMAZ). However, the response of ARGs in plastisphere with different biodegradability to freshwater-seawater shift is still unknown. In this study, ARG dynamics and associated microbiota on biodegradable poly (butyleneadipate-co-terephthalate) (PBAT) and non-biodegradable polyethylene terephthalate (PET) MPs were investigated through a simulated freshwater-seawater shift. The results exhibited that freshwater-seawater shift significantly influenced ARG abundance in plastisphere. The relative abundance of most studied ARGs decreased rapidly in plastisphere after they entered seawater from freshwater but increased on PBAT after MPs entered freshwater from seawater. Besides, the high relative abundance of multi-drug resistance (MDR) genes occurred in plastisphere, and the co-change between most ARGs and mobile genetic elements indicated the role of horizontal gene transfer on ARG regulation. Proteobacteria was dominant phylum in plastisphere and the dominant genera, such as Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Afipia, Gemmobacter and Enhydrobacter, were significantly associated with qnrS, tet and MDR genes in plastisphere. Moreover, after MPs entered new water environment, the ARGs and microbiota genera in plastisphere changed significantly and tended to converge with those in receiving water. These results indicated that MP biodegradability and freshwater-seawater interaction influenced potential hosts and distributions of ARGs, of which biodegradable PBAT posed a high risk in ARG dissemination. This study would be helpful for understanding the impact of biodegradable MP pollution on spread of antibiotic resistance in OMAZ.

Efficient bio-cementation between silicate tailings and biogenic calcium carbonate: Nano-scale structure and mechanism of the interface.

Biogenic calcium carbonate (bio-CaCO3) cementing tailings is an efficient technology to immobilize heavy metals in waste tailings. However, the underlying mechanism of interface cementation has not yet been clearly established, which limits the technological development. In this study, we used advanced techniques, including atomic force microscopy-based Lorentz contact resonance (AFM-LCR) spectroscopy, AFM-based nanoscale infrared (AFM-IR) spectroscopy, and solid-state nuclear magnetic resonance (ssNMR) spectroscopy, to reveal the structural, mechanical, and chemical properties of the interface on the nanoscale. Ureolytic bacteria produced bio-CaCO3 to fill in pore space and to bind cement tailings particles, which prevented the formation of leachate containing heavy metals. After cementation, a strong 40-300 nm thin interface was formed between the taillings and bio-CaCO3 particles. Unlike chemically synthesized CaCO3, bio-CaCO3 is strongly negatively charged, which gives it better adhesion ability. Fourier transform infrared (FTIR), AFM-IR, and 29Si ssNMR spectra indicated that the Si-OH and Si-O-Si groups on the silicate surface were converted to deprotonated silanol groups (≡Si-O-) at a high pH and they formed strong chemical bonds of Si-O-Ca on the interface through a Ca ion bridge. In addition, hydrogen bonding with Si-OH also played a role at the cementation interface. These findings provide the nano-scale interfacial structure and mechanism of bio-CaCO3 cementing silicate tailings and accelerate the development of tailings disposal technology.

Mechanisms for enhanced lignin humification with reduced organic matter loss by goethite in biogas residue composting.

In this study, effects of three iron (oxyhydr)oxides on the biogas residue composting, i.e., composting with goethite (CFe1), hematite (CFe2) or magnetite (CFe3), were investigated. Results showed that composting performance of CFe1 was much better than those of CFe2 and CFe3. Addition of goethite increased temperature of CFe1 and enhanced lignin humification. More than 31.49% of Fe(III) in goethite was reduced to amorphous Fe(II) during the composting, suggesting that goethite worked as electron acceptor for microbial metabolism and heat generation. The functional bacteria Chloroflexi and Actinobacteria, and genes encoding key enzymes (AA1 family), which play essential roles in humification of lignin, were enriched in CFe1. Besides, goethite reduced 10.96% organic matter (OM) loss probably by increasing the molecular size and aggregation of OM for its protection during the composting. This study shows that adding goethite is an efficient strategy to enhancing the humification of lignin-rich biowaste.

The persistently breaking trade-offs of three-decade plastic film mulching: Microplastic pollution, soil degradation and reduced cotton yield.

Plastic mulching has been extensively used for near 40 years in China because of the commonly recognized trade-offs between economic profit and adverse impacts. Whether the trade-offs is well kept after decades of mulching usage is poorly understood. In this study, microplastic (MP) pollution, soil physical properties, as well as the root traits and yields of cotton under plastic mulching for different years (up to 32 years) in northwest China were investigated. The results showed that average abundance of MPs in soil ranged from 28.00 to 1426.67 items kg-1 and increased with time whereas size of MPs decreased with time. With respect to soil physical properties, bulk density and penetration resistance significantly increased with mulching time while saturated hydraulic conductivity and mean weight diameter of water-stable aggregates in cotton fields decreased over time, indicating that long-term plastic mulching has irreversible adverse impact on soil function. The significantly increased soil penetration resistance and decreased hydraulic conductivity substantially restricted root growth and this finally led to remarkable reduction of seed cotton yield. It is urgent for the agricultural departments to take appropriate measures to mitigate the constantly increased negative trade-offs of plastic mulching in the arid areas.

Temporospatial nano-heterogeneity of self-assembly of extracellular polymeric substances on microplastics and water environmental implications.

Environmental behavior and ecotoxicity of microplastics (MPs) are significantly influenced by the omnipresent self-assembly of microbial extracellular polymeric substances (EPS) on them. However, mechanisms of EPS self-assembly onto MPs at nanoscale resolution and effects of aging are unclear. For the first time, temporospatial nano-heterogeneity of self-assembly of EPS onto fresh and one-year aged polypropylene (PP) MPs were investigated by atomic-force-microscopy-infrared-spectroscopy (AFM-IR). Natural aging caused high degree nanoscale fragmentation of MPs physically and chemically. Self-assembly of EPS on MPs was aging-dependent. Polysaccharides were assembled on MP surface faster than proteins. Initially, regardless of the fresh or aged MPs, polysaccharides and proteins, with the former being predominant, were successively and separately assembled to different nanospaces because of their competition for binding sites. More and more proteins and polysaccharides were superimposed on each other with assembly time due to intermolecular forces. The nanochemical textural analysis showed that the nano-heterogeneity of EPS assembly to MPs was clearly correlated with the aging-induced nanochemical and nanomechanical heterogeneity of MP surface. The spontaneous self-assembly of EPS with temporospatial nano-heterogeneity on MPs have multiple impacts on behavior, ecotoxicity and fate of MPs and their associated pollutants as well as other key ecological processes in aquatic environment.

Persistent versus transient, and conventional plastic versus biodegradable plastic? -Two key questions about microplastic-water exchange of antibiotic resistance genes.

The ubiquitous microplastics (MPs) in water environment play an important role in the dissemination of antibiotic resistance genes (ARGs) due to their exchange between floating MPs and receiving waters. However, whether the ARG exchange is persistent or transient and what are the differences in ARG exchange between conventional plastics and biodegradable plastics are the two key issues to be addressed. In this study, biodegradable PBAT and non-biodegradable PET MPs were chosen to explore the MP-water ARG exchange after the MPs floated to the receiving waters. The results demonstrated that the active exchange of ARGs between MPs and receiving waters occurred, which, however, were transient for most of ARGs. The relative abundance of ARGs both on the MPs and in the waters rapidly decreased to the initial or lower levels within 4 weeks. Approximately 25-50% (ARG subtype number ratio) of studied ARG subtypes were introduced into the receiving waters by MPs, and 35-65% of studied ARG subtypes went through fluctuation in terms of abundance on MPs and in the receiving water. ARGs tended to converge between MPs and the receiving waters with time. Furthermore, the ARG exchange between MPs and waters facilitated horizontal gene transfer (HGT). IntI1 and tnpA05 played the crucial roles in HGT, which was indicated by their correlated change with most ARGs; in contrast, tnpA04 showed the obvious lagging responses. The biodegradable MP of PBAT generally accumulated higher levels of most ARGs including multidrug resistant genes than the non-biodegradable MP of PET. The transient exchange of most ARGs between MPs and water implies that the on-off hitchhiking of ARGs on MPs in aquatic environment may not exert significant influence on ARG transmission. However, compared with the conventional plastics, the biodegradable MPs might pose much higher ARG dissemination risks due to the higher enrichment of ARGs particularly with people's ever-increasingly usage. Enough attention must be paid to this emerging issue.

Distinct fungal plastisphere across different river functional zones: A watershed scale study.

Fungi not only play important roles in biogeochemical processes but also can form biofilm on plastic debris. However, knowledge of structure composition and spatiotemporal pattern of fungal plastisphere on different kinds of plastic debris in river with specific usages, known as river functional zones, is still missing. In this study, we investigated the spatial distribution of the fungal plastisphere across a complete urban river with different functional zones (drinking, farm irrigation, aquaculture, and tail lake). Our research was performed based on both field residual plastic debris collection and a 30-day field in situ incubation experiments. Our study revealed that plastic debris enriched distinct fungal communities (including pathogenic fungi) significantly different from the surrounding water. Tracking the source of the fungi colonized on plastic debris suggested that the fungal taxa colonized on the different kinds of plastic debris were not from the surrounding water. Human activities had considerable effects on the fungal community structure on plastic debris, and the plastisphere fungal community structure strikingly varied across different river functional zones. Plastisphere may be used as an indicator for fungi biogeography and pathogenic fungi pollution in river with different functional zones. These findings are essential for ecological risk assessment and management decisions for pollution control of plastic debris and maintaining ecological health.

Ecotoxicological Effects of Microplastics on Bird Embryo Development by Hatching without Eggshell.

Microplastics are an emerging global pollutant type that poses a great health threat to animals due to their uptake and translocation in animal tissues and organs. Ecotoxicological effects of microplastics on the development of bird embryos are not known. The bird egg is a complete development and nutrition system, and the entire embryo development occurs in the eggshell. Therefore, a direct record of bird embryo development under the stress of pollutants such as microplastics is highly limited by the opaque eggshell in traditional hatching. In this study, the effects of microplastics on quail embryo development were visually monitored by hatching without an eggshell. The main steps include the cleaning and disinfection of fertilized eggs, the incubation before exposure, the short-term incubation after exposure, and the sample extraction. The results show that compared with the control group, the wet weight and body length of the microplastics-exposed group displayed a statistical difference and the liver proportion of the whole exposed group significantly increased. Additionally, we evaluated external factors that affect the incubation: temperature, humidity, egg rotation angle, and other conditions. This experimental method provides valuable information on the ecotoxicology of microplastics and a novel way to study the adverse effects of pollutants on the development of embryos.

Selectively enrichment of antibiotics and ARGs by microplastics in river, estuary and marine waters.

The partition of antibiotics and antibiotic resistant genes (ARGs) between the microplastics (MPs) and the surrounding water with various salinity are still unclear. In this study, we hypothesized that adsorption of antibiotics on MPs might cause a significant change of the structure of microbial communities, diversity and abundance of ARGs on MPs and this might be further affected by change of salinity. In this study, we investigated adsorption of four common antibiotics (sulfamerazine, tetracycline, chloramphenicol and tylosin) to polyethylene (PE) MPs in river, estuary and marine waters, and the differences of antibiotic resistant genes (ARGs) and bacterial communities on MPs and in the three waters. The results showed that MPs can enrich antibiotics, ARGs and microbes from the surrounding water. Elevated salinity could reduce adsorption of antibiotics to MPs and the abundance of ARGs. For example, MPs can concentrate more antibiotics and ARGs in the fresh river water than in the estuary and the marine waters. In addition, ARGs and bacterial communities on MPs at various salinity were significantly different under the pressure of four antibiotics. On MPs, sul1, sulA/folP-01, tetA, tetC, tetX and ermE increased significantly but a few new ARGs such as sulA/folP-01 and tetA appeared. The structure of the bacterial communities on MPs was different from the surrounding water since some bacteria species found on MPs were barely detected in the surrounding water while some genera on MPs vanished after exposure to antibiotics. As the antibiotics adsorbed and the ARGs on MPs decreased with the water salinity, the structure of the communities on MPs thus varied with salinity change. These findings are important to understand the effects of MPs on the transport, fate and ecological risk of antibiotics and ARGs in different aquatic environments.

Increased inheritance of structure and function of bacterial communities and pathogen propagation in plastisphere along a river with increasing antibiotics pollution gradient.

Plastic debris provides a stable substrate and novel ecological niche for microorganisms in the aquatic environment, which was referred to as "Plastisphere". Little is known about distribution patterns and responses of ecological function and structure of microbial communities in the plastisphere along rivers which usually have antibiotics pollution gradient. In this study, the differences in the community structure between the plastisphere and the planktonic bacteria, and their spatial variation of the community structure and function along a river with increased antibiotics pollution gradient was investigated at the watershed scale. The diversity of bacteria colonized on most plastic debris was higher than in surrounding water. Plastic debris could accumulate a higher abundance of some potential pathogens than surrounding water even at high antibiotics concentrations. The source tracking results showed that downstream plastisphere inherited much higher proportions of bacterial taxa from upstream than planktonic bacteria. About 92.3-99.7% of bacteria communities in downstream water were not from upstream but from the input of downstream human activities. On the contrary, high proportions of bacterial taxa in downstream plastisphere were closely connected to upstream. The plastisphere possesses higher ecological functional diversity than the planktonic bacteria. Seventy nine functional groups across plastisphere were predicted using functional annotation of prokaryotic taxa and only 65 functional groups were found in the planktonic bacteria. Plastisphere also acts as hotspot for biogeochemical cycling of nutrients such as N and S. Intensive human activities of urban and downstream agriculture and aquaculture had great effects on microbial community structure and functional groups of the Urumqi River. Pastisphere communities are much more resistant to human disturbance than planktonic bacteria. Compared to surrounding water, plastisphere increased inheritance from upstream microbial structure and function and also increased survival and propagation of pathogens in the downstream water with high concentrations of antibiotics.

Cadmium sorption by EPSs produced by anaerobic sludge under sulfate-reducing conditions.

This study showed that EPS produced by anaerobic sludge under sulfate-reducing bacteria was effective in removing Cd(2+) from solution. The sorption data could be described by the Freundlich isotherm model. The q(m) derived from the Langmuir isotherm model was up to 2720 mg/g EPSs. The presence of copper and nickel ions had significantly reduced the cadmium sorption by EPS and the presence of zinc had little effect. Analysis of FTIR spectra demonstrated that C-O-C of polysaccharides at 1150-1030 cm(-1), group of the amide(I), CH(2) group of the lipids, carboxyl and -OH groups of proteins and polysaccharides were involved in cadmium binding, of which the -OH groups and the C-O-C group of polysaccharides played a major role in cadmium sorption by EPSs.

Extracellular polymeric substances buffer against the biocidal effect of H2O2 on the bloom-forming cyanobacterium Microcystis aeruginosa.

H2O2 is an emerging biocide for bloom-forming cyanobacteria. It is important to investigate the H2O2 scavenging ability of extracellular polymeric substances (EPS) of cyanobacteria because EPS with strong antioxidant activity may "waste" considerable amounts of H2O2 before it kills the cells. In this study, the buffering capacity against H2O2 of EPS from the bloom-forming cyanobacterium Microcystis aeruginosa was investigated. IC50 values for the ability of EPS and vitamin C (VC) to scavenge 50% of the initial H2O2 concentration were 0.097 and 0.28 mg mL(-1), respectively, indicating the higher H2O2 scavenging activity of EPS than VC. Both proteins and polysaccharides are significantly decomposed by H2O2 and the polysaccharides were more readily decomposed than proteins. H2O2 consumed by the EPS accounted for 50% of the total amount of H2O2 consumed by the cells. Cell growth and photosynthesis were reduced more for EPS-free cells than EPS coated cells when the cells were treated with 0.1 or 0.2 mg mL(-1) H2O2, and the maximum photochemical efficiency Fv/Fm of EPS coated cells recovered to higher values than EPS-free cells. Concentrations of H2O2 above 0.3 mg mL(-1) completely inhibited photosynthesis and no recovery was observed for both EPS-free and EPS coated cells. This shows that EPS has some buffering capacity against the killing effect of H2O2 on cyanobacterial cells. Such a strong H2O2 scavenging ability of EPS is not favorable for killing bloom-forming cyanobacteria. The high H2O2 scavenging capacity means considerable amounts of H2O2 have to be used to break through the EPS barrier before H2O2 exerts any killing effects on the cells. It is therefore necessary to determine the H2O2 scavenging capacity of the EPS of various bloom-forming cyanobacteria so that the cost-effective amount of H2O2 needed to be used for killing the cyanobacteria can be estimated.

Effects of irradiation and pH on fluorescence properties and flocculation of extracellular polymeric substances from the cyanobacterium Chroococcus minutus.

Microbial extracellular polymeric substances (EPS) may flocculate or be decomposed when environmental factors change, which significantly influences nutrient cycling and transport of heavy metals. However, little information is available on the stability of EPS in natural environments. Fluorescence and flocculation properties of EPS from Chroococcus minutus under different irradiation and pH conditions were studied. Two aromatic protein-like fluorescence peaks and one tyrosine protein-like peak were identified from the excitation-emission-matrix (EEM) fluorescence spectra of EPS. UVB (ultraviolet B) and solar irradiation increased the fluorescence intensity of all the three peaks while UVC (ultraviolet C) irradiation had little effect. EPS formed unstable flocs after exposure to UV (ultraviolet) irradiation and formed stable flocs under solar irradiation. EPS were prone to flocculation under highly acidic conditions and minimal fluorescence of peaks was observed. The fluorophores in EPS were relatively stable under neutral and alkaline conditions. These findings are helpful for understanding the behavior of EPS in aquatic environments and their role in biogeochemical cycles of the elements.

Biosorption of Hg(II) onto goethite with extracellular polymeric substances.

This study characterized the interactions of goethite, EPS from cyanobacterium Chroococcus sp. and Hg(II) using excitation emission matrix (EEM) spectra and adsorption isotherms. Three protein-like fluorescence peaks were noted to quench in the presence of Hg(II). The estimated conditional stability constant (logKa) and the binding constant (logKb) of the studied EPS-Hg(II) systems ranged 3.84-4.24 and 6.99-7.69, respectively. The proteins in EPS formed stable complex with Hg(II). The presence of proteins of Chroococcus sp. enhanced the adsorption capacity of Hg(II) on goethite; therefore, the goethite-EPS soil is a larger Hg(II) sink than goethite alone soil. Biosorption significantly affects the mobility of Hg(II) in goethite soils.

Desorption of Hg(II) and Sb(V) on extracellular polymeric substances: effects of pH, EDTA, Ca(II) and temperature shocks.

Extracellular polymeric substances (EPS) existed ubiquitously in biological systems affect the mobility and availability of heavy metals in the environments. The adsorption-desorption behaviors of Hg(II) and Sb(V) on EPS were investigated. The sorption rates follow Sb(V) > Hg(II), and the desorption rates follow reverse order. Applications of ethylene diamine tetraacetic acid (EDTA), Ca(II) and pH shocks affect desorption rates and desorbed quantities of Hg(II) from EPS-Hg complex. Temperature shock minimally affects the desorption rate of Hg(II). Conversely, the EPS-Sb complex is stable subjected to EDTA, Ca(II), temperature or pH shocks. The excitation-emission matrix (EEM) fluorescence spectroscopy and fast-Fourier (FT-IR) analysis showed that Hg(II) and Sb(V) principally interacted with polysaccharides and protein-like compounds in the EPS, respectively. The EPS-Hg complex presents a time bomb that may release high levels of Hg(II) in short time period under environmental shocks.

Binding of phenanthrene to extracellular polymeric substances (EPS) from aerobic activated sludge: a fluorescence study.

Extracellular polymeric substances (EPS) are the essential components of activated sludge for removal of pollutants from wastewater. Limited information is available on the binding constants and binding mode of organic pollutants to EPS. In the present study, binding of phenanthrene (PHE) to extracellular polymeric substances (EPS) from aerobic activated sludge was investigated using fluorescence spectroscopy. Two protein-like fluorescence peaks (Ex/Em=225nm/340nm for peak A; Ex/Em=275-280nm/338nm for peak B) were identified in the three-dimensional excitation-emission matrix (3DEEM) fluorescence spectroscopy of the EPS. The fluorophores in EPS were clearly quenched by PHE and the quenching processes were static. The quenching constants (lnK(a)) were in the range of 11.27-13.82M(-1) and the binding constants (logK(b)) in the range of 6.11-8.98M(-1). The binding site number increased with increasing temperature. The corresponding thermodynamic parameters DeltaG, DeltaH and DeltaS were calculated. The interaction of the fluorophores in EPS and PHE is spontaneous and exothermic. The binding of EPS with PHE is dominated by the hydrophobic interactions. Fluorophore A has stronger hydrophobic interaction with PHE than fluorophore B.

Complexation between Hg(II) and biofilm extracellular polymeric substances: an application of fluorescence spectroscopy.

The three-dimensional excitation emission matrix (EEM) fluorescence spectroscopy was employed to investigate the interaction of extracellular polymeric substances (EPS) from natural biofilm with Hg(II). The EEM spectra demonstrated that EPS with molecular weight over 14 kDa had two protein-like fluorescence peaks. The fluorescence intensity at both peaks was strongly dependent on the solution pH in the absence and presence of Hg(II), with the maximal fluorescence intensity at neutral pH. Fluorescence of both protein-like peaks was significantly quenched by Hg(II). The values of conditional stability constants (log K(a)=3.28-4.48) derived from modified Stern-Volmer equation are approximate to those for humic substances and dissolved organic matter (DOM), indicating that fluorescent components in EPS have strong binding capacity for Hg(II). Our findings suggest that EPS from biofilm is a class of important organic ligands for complexation with Hg(II) and may significantly affect the chemical forms, mobility, bioavailability and ecotoxicity of heavy metals in the aquatic environment.