Enhanced periphyton biodegradation of endocrine disrupting hormones and microplastic: Intrinsic reaction mechanism, influential humic acid and microbial community structure elucidation.

Endocrine-disrupting compounds (EDCs), as well as microplastics, have drawn global attention due to their presence in the aquatic ecosystem and persistence in wastewater treatment plants (WWTPs). In the present study, for simultaneous bio-removal of two EDCs, 17α-ethinylestradiol (EE2), bisphenol A (BPA), and a microplastic, polypropylene (PP) four kinds of periphytic biofilms were employed. Additionally, the effect of humic acid (HA) on the removal efficacy of these biofilms was evaluated. It was observed that EE2 and BPA (0.2 mg L-1 each) were completely (∼100%) removed within 36 days of treatment; and the biodegradation of EE2, BPA, and PP was significantly enhanced in the presence of HA. Biodegradation of EE2 and BPA was evaluated through Ultra-high performance liquid chromatography (UHPLC), and Gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) was used to determine the mechanism of degradation. Gel permeation chromatography (GPC) and SEM had validated the biodegradation of PP (5.2-14.7%). MiSeqsequencing showed that the community structure of natural biofilm changed after the addition of HA, as well as after the addition of EDCs and PP. This change in community structure might be a key factor regarding variable biodegradation percentages. The present study revealed the potential of periphytic biofilms for the simultaneous removal of pollutants of different chemical natures, thus provides a promising new method for wastewater treatment applications.

Multifunctional Periphytic Biofilms: Polyethylene Degradation and Cd2+ and Pb2+ Bioremediation under High Methane Scenario.

Priority pollutants such as polyethylene (PE) microplastic, lead (Pb2+), and cadmium (Cd2+) have attracted the interest of environmentalists due to their ubiquitous nature and toxicity to all forms of life. In this study, periphytic biofilms (epiphyton and epixylon) were used to bioremediate heavy metals (HMs) and to biodegrade PE under high (120,000 ppm) methane (CH4) doses. Both periphytic biofilms were actively involved in methane oxidation, HMs accumulation and PE degradation. Epiphyton and epixylon both completely removed Pb2+ and Cd2+ at concentrations of 2 mg L-1 and 50 mg L-1, respectively, but only partially removed these HMs at a relatively higher concentration (100 mg L-1). Treatment containing 12% 13CH4 proved to be most effective for biodegradation of PE. A synergistic effect of HMs and PE drastically changed microbial biota and methanotrophic communities. High-throughput 16S rRNA gene sequencing revealed that Cyanobacteria was the most abundant class, followed by Gammaproteobacteria and Alphaproteobacteria in all high-methane-dose treatments. DNA stable-isotope probing was used to label 13C in a methanotrophic community. A biomarker for methane-oxidizing bacteria, pmoA gene sequence of a 13C-labeled fraction, revealed that Methylobacter was most abundant in all high-methane-dose treatments compared to near atmospheric methane (NAM) treatment, followed by Methylococcus. Methylomonas, Methylocystis, Methylosinus, and Methylocella were also found to be increased by high doses of methane compared to NAM treatment. Overall, Cd+2 had a more determinantal effect on methanotrophic activity than Pb2+. Epiphyton proved to be more effective than epixylon in HMs removal and PE biodegradation. The findings proved that both epiphyton and epixylon can be used to bioremediate HMs and biodegrade PE as an efficient ecofriendly technique under high methane concentrations.

Enhanced Adsorptive Bioremediation of Heavy Metals (Cd2+, Cr6+, Pb2+) by Methane-Oxidizing Epipelon.

Cadmium (Cd), chromium (Cr) and lead (Pb) are heavy metals that have been classified as priority pollutants in aqueous environment while methane-oxidizing bacteria as a biofilter arguably consume up to 90% of the produced methane in the same aqueous environment before it escapes into the atmosphere. However, the underlying kinetics and active methane oxidizers are poorly understood for the hotspot of epipelon that provides a unique micro-ecosystem containing diversified guild of microorganisms including methane oxidizers for potential bioremediation of heavy metals. In the present study, the Pb2+, Cd2+and Cr6+ bioremediation potential of epipelon biofilm was assessed under both high (120,000 ppm) and near-atmospheric (6 ppm) methane concentrations. Epipelon biofilm demonstrated a high methane oxidation activity following microcosm incubation amended with a high concentration of methane, accompanied by the complete removal of 50 mg L-1 Pb2+ and 50 mg L-1 Cd2+ (14 days) and partial (20%) removal of 50 mg L-1 Cr6+ after 20 days. High methane dose stimulated a faster (144 h earlier) heavy metal removal rate compared to near-atmospheric methane concentrations. DNA-based stable isotope probing (DNA-SIP) following 13CH4 microcosm incubation revealed the growth and activity of different phylotypes of methanotrophs during the methane oxidation and heavy metal removal process. High throughput sequencing of 13C-labelled particulate methane monooxygenase gene pmoA and 16S rRNA genes revealed that the prevalent active methane oxidizers were type I affiliated methanotrophs, i.e., Methylobacter. Type II methanotrophs including Methylosinus and Methylocystis were also labeled only under high methane concentrations. These results suggest that epipelon biofilm can serve as an important micro-environment to alleviate both methane emission and the heavy metal contamination in aqueous ecosystems with constant high methane fluxes.

Periphytic biofilm: An innovative approach for biodegradation of microplastics.

Microplastics (MPs) have been gaining the attention of environmental researchers since the 1960s anecdotal reports of plastic entanglement and ingestion by marine creatures. Due to their increasing accretion in aquatic environments, as well as resistance towards degradation, marine litter research has focused on microplastics more recently. In the present study, a relatively new method of biodegradation was implemented for the biodegradation of three structurally different MPs i.e. polypropylene (PP), polyethylene (PE) and polyethylene terephthalate (PET). Periphytic biofilm was used for this purpose in various backgrounds of carbon sources (glucose, peptone, and glucose and peptone). Biodegradation of MPs was estimated in terms of weight loss. It was observed that the addition of glucose enhanced the biodegradation of MPs by periphyton biofilm for all MPs (from 9.52%-18.02%, 5.95%-14.02% and 13.24-19.72% for PP, PE and PET respectively) after 60 days compared to natural biofilm alone. To the contrary, peptone, and glucose and peptone together, were inhibitory. Biodegradation was further confirmed by morphological changes observed using SEM, FTIR spectra and GPC lent further support to the results whereby new peaks appeared along with reduction in old peaks and decrease in peak intensities. MiSeq sequencing shows that Deinococcus-thermus > Proteobacteria > Cyanobacteria are the dominant phyla in natural biofilms, and their relative abundances increase after the addition of glucose. However, the abundances shifted to Deinococcus-thermus > Cyanobacteria > Firmicutes > Bacteroidetes, when the biofilms were treated with either peptone alone, or with glucose and peptone together. Therefore, the change in biodegradation capability might also be due to the change in the microbial community structures after addition of the C-sources. These experiments provide an innovative approach towards effective biodegradation of MPs using a relatively new environment-friendly method.

Periphyton biofilms: A novel and natural biological system for the effective removal of sulphonated azo dye methyl orange by synergistic mechanism.

Due to their large scale use, azo dyes are adversely affecting aquatic fauna and flora as well as humans. The persistent nature of sulphonated azo dyes makes them potential ecotoxic hazards. The aim of the present study was to employ a proficient, locally available biomaterial, viz. periphyton (i.e. epiphyton, epilithon or metaphyton), for removal of the azo dye, methyl orange (MO). Results showed that the periphytic biofilms are capable of completely removing comparatively high concentrations (up to 500 mg L-1) of MO from wastewater. The removal of MO occurs by a synergistic mechanism involving bioadsorption and biodegradation processes. The adsorption of MO by periphyton can be described by pseudo-second order kinetics. Elovich and intraparticle diffusion models as well as Langmuir equations fit well to the MO adsorption process. FTIR analysis of MO and its metabolites demonstrated biotransformation into simpler compounds within 72 h. GC-MS/MS analysis showed the conversion of MO into simpler compounds such as phenol, ethyl acetate and acetyl acetate. The results indicated that periphyton is a promising biomaterial for the complete removal of MO from wastewater and that the treatment process has the potential for in situ removal of MO at contaminated sites.

Evaluating role of immobilized periphyton in bioremediation of azo dye amaranth.

The aim of this study was to evaluate the bioremediation capabilities of three kinds of periphyton (i.e. epiphyton, metaphyton and epilithon) immobilized in bioreactors to decolorize and biodegrade the sulphonated azo dye, amaranth. Results showed that periphyton dominated by phyla including Cyanobacteria, Proteobacteria and Bacteroidetes. Complete removal of dye was shown by all the biofilms periphyton (epiphyton showed highest removal efficacy) over a range of initial concentrations (50-500mgL-1) within 84h at pH 7 and 30°C. Biodegradation of amaranth was confirmed through FTIR and HPLC and the biodegradation pathways were detected by GC-MS/MS analysis. The azo bonds in the amaranth were successfully broken by periphyton and amaranth was converted to non-toxic, aliphatic compounds including isobutene, acetyl acetate and ethyl acetate. The results showed the potential application of immobilized periphyton at industrial scale for the removal of azo dyes from wastewater containing azo dye amaranth.

The adsorption process during inorganic phosphorus removal by cultured periphyton.

To explain the detailed process involved in phosphorus removal by periphyton, the periphyton dominated by photoautotrophic microorganisms was employed in this study to remove inorganic phosphorus (Pi) from wastewater, and the removal kinetics and isotherms were then evaluated for the Pi removal process. Results showed that the periphyton was capable of effectively removing Pi that could completely remove the Pi in 24 h at an initial Pi concentration of 13 mg P L(-1). Furthermore, the Pi removal process by the periphyton was dominated by adsorption at initial stage (~24 h), which involved physical mechanistic process. However, this Pi adsorption process was significantly influenced by environmental conditions. This work provides an insight into the understanding of phosphorus adsorption by periphyton or similar microbial aggregates.

In situ bioremediation of surface waters by periphytons.

Environmentally benign and sustainable biomeasures have become attractive options for the in situ remediation of polluted surface waters. In this paper, we review the current state of reported experiments utilizing naturally occurring periphyton. These are microbial communities consisting of heterotrophic and photoautotrophic microorganisms that are reportedly capable of remediating surface waters which suffer from pollution due to a variety of contaminants. In our review, we focus on four aspects of bioremediation: multiple contaminant removal, the processes involved in contaminant removal, successful cell immobilization technologies and finally, the consideration of safety in aquaculture. It has been noted that recent developments in immobilization technologies offer a fresh approach facilitating the application of periphyton. The use of periphyton biofilm overcomes several disadvantages of single species microbial aggregates. The inclusion of periphyton, as a stable micro-ecosystem, is a promising in situ strategy to restore decimated surface water ecosystems.