Plastic pollution is ubiquitous throughout the marine environment, with microplastic (i.e. <5 mm) contamination a global issue of emerging concern. The lack of universally accepted methods for quantifying microplastic contamination, including consistent application of microscopy, photography, an spectroscopy and photography, may result in unrealistic contamination estimates. Here, we present and apply an analysis workflow tailored to quantifying microplastic contamination in marine waters, incorporating stereomicroscopic visual sorting, microscopic photography and attenuated total reflectance (ATR) Fourier transform infrared (FTIR) spectroscopy. The workflow outlines step-by-step processing and associated decision making, thereby reducing bias in plastic identification and improving confidence in contamination estimates. Specific processing steps include (i) the use of a commercial algorithm-based comparison of particle spectra against an extensive commercially curated spectral library, followed by spectral interpretation to establish the chemical composition, (ii) a comparison against a customised contaminant spectral library to eliminate procedural contaminants, and (iii) final assignment of particles as either natural- or anthropogenic-derived materials, based on chemical type, a compare analysis of each particle against other particle spectra, and physical characteristics of particles. Applying this workflow to 54 tow samples collected in marine waters of North-Western Australia visually identified 248 potential anthropogenic particles. Subsequent ATR-FTIR spectroscopy, chemical assignment and visual re-inspection of photographs established 144 (58%) particles to be of anthropogenic origin. Of the original 248 particles, 97 (39%) were ultimately confirmed to be plastics, with 85 of these (34%) classified as microplastics, demonstrating that over 60% of particles may be misidentified as plastics if visual identification is not complemented by spectroscopy. Combined, this tailored analysis workflow outlines a consistent and sequential process to quantify contamination by microplastics and other anthropogenic microparticles in marine waters. Importantly, its application will contribute to more realistic estimates of microplastic contamination in marine waters, informing both ecological risk assessments and experimental concentrations in effect studies.

In this study, we examined the removal of arsenite (As(III)) and arsenate (As(V)) by perilla leaf-derived biochars produced at 300 and 700 °C (referred as BC300 and BC700) in aqueous environments. Results revealed that the Langmuir isotherm model provided the best fit for As(III) and As(V) sorption, with the sorption affinity following the order: BC700-As(III) > BC700-As(V) > BC300-As(III) > BC300-As(V) (QL = 3.85–11.01 mg g⁻¹). In general, As removal decreased (76–60%) with increasing pH from 7 to 10 except for the BC700-As(III) system, where notably higher As removal (88–90%) occurred at pH from 7 to 9. Surface functional moieties contributed to As sequestration by the biochars examined here. However, significantly higher surface area and aromaticity of BC700 favored a greater As removal compared to BC300, suggesting that surface complexation/precipitation dominated As removal by BC700. Arsenic K-edge X-ray absorption near edge structure (XANES) spectroscopy demonstrated that up to 64% of the added As(V) was reduced to As(III) in BC700- and BC300-As(V) sorption experiments, and in As(III) sorption experiments, partial oxidation of As(III) to As(V) occurred (37–39%). However, XANES spectroscopy was limited to precisely quantify As binding with sulfur species as As2S3-like phase. Both biochars efficiently removed As from natural As-contaminated groundwater (As: 23–190 μg L⁻¹; n = 12) despite in the presence of co-occurring anions (e.g., CO3²⁻, PO4³⁻, SO4²⁻) with the highest levels of As removal observed for BC700 (97–100%). Overall, this study highlights that perilla leaf biochars, notably BC700, possessed the greatest ability to remove As from solution and groundwater (drinking water). Significantly, the integrated spectroscopic techniques advanced our understanding to examine complex redox transformation of As(III)/As(V) with biochar, which are crucial to determine fate of As on biochar in aquatic environments.

Microplastics (MPs) are generated from plastic and have negative impact to our environment due to high level of fragmentation. They can be originated from various sources in different forms such as fragment, fiber, foam and so on. For detection of MPs, many techniques have been developed with different functions such as microscopic observation, density separation, Raman and FTIR analysis. Besides, due to ingestion of MPs by wide range of marine species, research on the effect of this pollution on biota as well as human is vital. Therefore, we comprehensively reviewed the occurrence and distribution of MPs pollution in both marine and freshwater environments, including rivers, lakes and wastewater treatment plants (WWTPs). For future studies, we propose the development of new techniques for sampling MPs in aquatic environments and biota and recommend more research regarding MPs release by WWTPs.

Microplastics (<5 mm) have often been studied under in-vitro conditions where plastics have been investigated in isolation. However, in the natural environment microplastics readily form agglomerates conferring the particles with properties different to their pristine counterparts. Here, we examined the interaction of exopolymers with polystyrene nanoplastics and microplastics. Formation of plastic agglomerates was examined using simulated sea surface conditions. Flow cytometry coupled with microscopy revealed that nano- and microplastic particle spheres form agglomerates in seawater with a mucilagenous material and an associated microbial community. To characterise this material, differential staining methods revealed it to be glycoprotein in composition. Exposing increasing concentrations of a marine bacterial glycoprotein EPS to nano- or microplastics revealed that these types of polymers contribute to the formation and abundance of plastic agglomerates. This work highlights the importance of EPS on the fate of plastic and future research should take this into account when evaluating the impact of plastics.

Shipping is recognised as an unintentional efficient pathway for spreading non-native species, harmful organisms and pathogens. In 2004, a unique IMO Convention was adopted to control and minimize this transfer in ship's ballast water. This Convention entered into force on 8th September 2017. However, unlikely the majority of IMO Conventions, the Ballast Water Management Convention requires ships to comply with biological standards (e.g. concentration of organisms per unit of volume in ballast water discharges). This study aimed to apply different techniques developed to measure concentrations of viable phytoplankton in natural and treated ballast water samples and compare them with the established flow cytometry method and vital staining microscopy. Samples were collected in the English Channel over one year and on-board during ballast water shipboard efficacy tests. Natural abundance of live phytoplankton varied from 23% to 89% of the total, while for cells larger than 10 μm (a size defined by the BWM Convention) the percentage varied from 3% to 60%. An overall good correlation was seen between the measurements taken with the two fluorometers and in comparison with the flow cytometry analysis, as found in previous studies. Analysis of treated ballast water samples showed a large variation in the number of viable cells, however indicating a low level of risk on all occasions for regulatory purposes. One of the key aspects to bear in mind when sampling and analysing for compliance is to be aware of the limitations of each technique.

Large seaweed cultivation has proven an effective means to inhibit harmful microalgae at experimental scales and battle eutrophication in Chinese coastal waters, but essentially there is a lack of field-scale studies to explore the underlying mechanism. Here we present a 1.5-year-long time series of particulate organic matter (POM) and settling particulate matter (SPM) concentrations from an integrated aquaculture of Gracilaria lemaneiformis off the coast of Nan'ao Island, South China from April 2014 to August 2015. The microscopic examination and geochemical characteristics show that the POM mainly consisted of microalgae. The mean POM concentration increased 99.8%, 71.2%, 45.8% and 111.9% at the four sampling sites during the non-cultivation period, while decreased 25.5%, 17.3%, 12.2% and 20.3%, respectively, during the seaweed cultivation period. These results suggest that the large scale seaweed cultivation can remove excess nutrients and inhibit microalgal growth, thereby contributing to the improvement of coastal marine aquaculture environment.

The applicability of spectral analysis in detection of freshwater metal contamination was assessed by developing and testing a novel hyperspectral imaging (HSI) application for aquatic insect larvae (Trichoptera: Hydropsychidae). Larvae were first exposed to four different cadmium (Cd) concentrations: 0, 1, 10, and 100 μg L⁻¹ for 96 h. Individual larvae were then preserved in ethanol, inspected with microscopy for the number of anomalies in larval gills, and imaged by hyperspectral camera operating with wavebands between 500 and 850 nm. Three additional larvae from each exposure were analyzed for tissue Cd concentration. Although the larval tissue Cd concentrations correlated positively with actual water concentrations, the toxicity response of larvae, i.e., frequency of gill abnormalities, did not differ among the Cd concentrations. In contrast, hyperspectral imaging data indicated some concentration-response relationship of larval spectral properties to the Cd exposure, but it was too weak for reliable automatic distinction between exposed and unexposed larvae. In this pilot study a workflow for data processing for a novel application of hyperspectral imaging was developed. Based on the results of this preliminary study, the workflow in the imaging process will be optimized and its potential for detecting metal contamination of aquatic environments reassessed.

The corrosion products usually found on outdoor bronzes are generated by the interaction between the metal alloy and the atmospheric pollutants. To protect the external surface of bronzes, different organic materials (natural or synthetic) can be applied, creating over time a patina consisting of a complex mixture of inorganic and organic degraded components. The correct chemical characterization of patina constituents is fundamental to define the state of conservation of a metal artwork and address proper restoration actions. In this paper, we evaluated the potentialities of near-infrared (NIR) reflectance microscopy (4000–7500 cm⁻¹) as complementary method to mid-infrared (MIR) analyses for the characterization of bronze patinas. Although NIR spectroscopy has been already used in the field of heritage science, its application for the characterization of bronze patinas is almost unexplored. In this paper, several corrosion products usually found on the surface of outdoor bronze sculptures were synthesized, characterized, and submitted to the NIR-MIR total reflection analysis to build up a reference spectral database. We devoted particular attention to the NIR features of copper hydroxychlorides, such as atacamite and paratacamite, which have not been studied in detail up to now. A selection of organic-based formulations, commonly used by restorers to protect the bronze surface against the outdoor aggressive environment, were also considered as references. Successively, NIR-MIR reflectance microscopy was successfully employed for the analysis of patina micro-samples collected from the bronze statues of the Neptune Fountain (sixteenth century) located in Bologna. The obtained results demonstrate the ability of NIR spectroscopy to identify organic and inorganic patina constituents, even in mixtures. In addition, the study can be considered as a proof of concept for the possible future application of the technique for in situ diagnostic campaigns on bronze sculptures.

The Cyanobacterium Oculatella subterranea Zammit, Billi, Albertano inhabits hypogea and stone caves and is a pioneer of different stone substrata. In this study, a strain isolated from the House of Marco Castricio (Archaeological Park of Pompeii, Italy) was identified by a polyphasic approach and used for an in vitro colonization test to verify the influence of the substrate on the biofilm architecture. Fine structure of O. subterranea microbial mats was revealed as well as filaments orientation toward light source. This aim has been achieved through confocal laser scanner microscope microscopy and computer image analysis. Moreover, bioreceptivity of five different substrates, commonly retrieved in archaeological sites of Campania, was assessed for O. subterranea. Our results show that the three-dimensional structure of O. subterranea microbial mats is poorly affected by physical and geochemical features of substrates: in fact, the porous architecture of its biofilm was preserved, independently of the materials. On the other hand, the area/perimeter ratio relative to the O. subterranea growth on tuff, brick, and porphyry showed significant differences, indicating dissimilar levels of bioreceptivity of the three substrates.

Simultaneous oxidation of Hg⁰ and NH₃-SCR of NO by catalyst is one of the key methods for co-purification of coal-fired flue gas. Till now, the interaction between the oxidation of Hg⁰ and NH₃-SCR of NO and its mechanism have not clarified. In this study, a series of nanophase Ce ₓ Zr y Mn z O₂ was prepared for the simultaneous oxidation of Hg⁰ and NH₃-SCR of NO at low temperature. The catalysts were characterized using surface area analysis, X-ray diffraction, temperature-programmed techniques, and several types of microscopy and spectroscopy. The experimental results indicated that the Ce₀.₄₇Zr₀.₂₂Mn₀.₃₁O₂ exhibited superior Hg⁰ removal efficiency (> 99%) and NO conversion efficiency (> 90%) even at 150 °C, and it also exhibited a good durability in the presence of SO₂ and H₂O. The excellent performance of Ce₀.₄₇Zr₀.₂₂Mn₀.₃₁O₂ on co-purifying Hg⁰ and NO was due to the stronger synergistic effects of Ce-Zr-Mn in Ce₀.₄₇Zr₀.₂₂Mn₀.₃₁O₂ than that of the others, which was illustrated by the characterization results of XPS, XRD, and FT-IR. Moreover, it was found that the NO conversion of Ce₀.₄₇Zr₀.₂₂Mn₀.₃₁O₂ could be slightly influenced by Hg⁰ and was decreased about 4% to the max, while that of Hg⁰ could rarely be affected by the selected catalytic reduction process of NO. It might be due to the co-purification mechanism of NO and Hg⁰. The mechanism of the simultaneous oxidation of Hg⁰ and NH₃-SCR of NO was mainly due to the synergetic effect on the mobility of surface oxygen and the activation of lattice oxygen of Ce₀.₄₇Zr₀.₂₂Mn₀.₃₁O₂. The effect of the oxidation of Hg⁰ on the NH₃-SCR of NO was mainly due to the absorbed Hg⁰/Hg²⁺ on the surface of Ce₀.₄₇Zr₀.₂₂Mn₀.₃₁O₂, which attenuated the formation of NH₃₍ₐd₎, −NH₂₍ₐd₎, and NH₄⁺ on its acid sites. Similarly, the NH₃-SCR of NO process could hardly influence the oxidation of Hg⁰ when NO and Hg⁰ were co-purified.