Electrophoretic deposition is a straightforward method for synthesizing high-quality photoanodes. We present a protocol for synthesizing a TiO2-modified boron-doped diamond photoanode (BDD/TiO2) via electrophoretic deposition, detailing the chemical and electrochemical treatments applied to the bare BDD electrode prior to use. We provide a step-by-step guide for performing photoelectrochemical characterization under both dark and light conditions and describe an optical technique for investigating band-gap energy. For complete details on the use and execution of this protocol, please refer to Quilumbaquin et al.1.

Increasing plastic waste is a critical global challenge to ecological and human health requiring focused solutions to reduce omnipresent plastic pollution in the environment. While recycling has been touted as one solution to counter plastic waste and resource utilization, it has been largely ineffective in offsetting the impact of rising global plastic production of more than 400 million metric tonnes annually, due to low global recycling rates of only 9%. Over three decades since implementing plastic resin codes, recycling has favoured thermoplastics, neglecting thermoset plastics. There is a constant need to enhance overall recycling efficiency by exploring advanced methods, as enormous gaps exist in fully unlocking the potential of plastic recycling. We identify critical gaps associated with plastic waste recycling and its potential environmental impacts. We discuss substantial progress in recycling technology, designs-for-recyclability with controlled chemical use, and economic incentives to expand markets for recycled plastics and to curb plastic leakage into the environment. Additionally, we highlight some emerging strategies and legally binding international policy instruments, such as the Global Plastics Treaty that require further development to reduce plastic waste and improve plastic recyclability.

Research on fungi under anaerobic conditions is limited but crucial for understanding their ecological and pathological impacts. Here, we present a protocol for enriching, isolating, and characterizing anaerobic fungi from environmental and clinical samples. We also describe steps for evaluating the anaerobic growth potential and drug susceptibility of fungal pathogens. This protocol can contribute to the need for initiating effective antifungal therapy to address and manage fungal infections or mycosis in oxygen-limited environments. For complete details on the use and execution of this protocol, please refer to Yadav et al.1.

The stop signal is produced in response to negative experiences at the food source and inhibits honey bee (Apis mellifera) waggle dancing. Here, we present a protocol for measuring the effects of an inhibitory signal associated with danger on honey bee dopamine levels. We describe steps for observing honey bee colonies, training them with artificial nectar, and simulating hornet attacks. We then detail procedures for recording waggle dancing and stop signals and measuring brain dopamine levels during different treatments. For complete details on the use and execution of this protocol, please refer to Dong et al.1.

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When plants are subjected to drought, it is impossible to obtain xylem sap for subsequent biochemical and molecular analysis using root pressure exudate, the conventional approach. Here, we present a protocol for collecting xylem sap from drought-treated tomato plants using a pressure chamber. We describe steps for how to prepare plants, apply drought, and use the pressure chamber to collect the xylem sap. Using this technique, one can obtain 500-700 μL of xylem sap in just 5-7 min. For complete details on the use and execution of this protocol, please refer to Alexou and Peuke.1.

Secondary inorganic aerosols play an important role in air pollution and climate change, and their formation modulates the atmospheric deposition of reactive nitrogen (including oxidized and reduced nitrogen), thus impacting the nitrogen cycle. Large-scale and long-term analyses of secondary inorganic aerosol formation based on model simulations have substantial uncertainties. Here we improve constraints on secondary inorganic aerosol formation using decade-long in situ observations of aerosol composition and gaseous precursors from multiple monitoring networks across the United States. We reveal a shift in the secondary inorganic aerosol formation regime in the rural United States between 2011 and 2020, making rural areas less sensitive to changes in ammonia concentrations and shortening the effective atmospheric lifetime of reduced forms of reactive nitrogen. This leads to potential increases in reactive nitrogen deposition near ammonia emission hotspots, with ecosystem impacts warranting further investigation. Ammonia (NH3), a critical but not directly regulated precursor of fine particulate matter in the United States, has been increasingly scrutinized to improve air quality. Our findings, however, show that controlling NH3 became significantly less effective for mitigating fine particulate matter in the rural United States. We highlight the need for more collocated aerosol and precursor observations for better characterization of secondary inorganic aerosols formation in urban areas.

Imagery has become one of the main data sources for investigating seascape spatial patterns. This is particularly true in deep-sea environments, which are only accessible with underwater vehicles. On the one hand, using collaborative web-based tools and machine learning algorithms, biological and geological features can now be massively annotated on 2D images with the support of experts. On the other hand, geomorphometrics such as slope or rugosity derived from 3D models built with structure from motion (sfm) methodology can then be used to answer spatial distribution questions. However, precise georeferencing of 2D annotations on 3D models has proven challenging for deep-sea images, due to a large mismatch between navigation obtained from underwater vehicles and the reprojected navigation computed in the process of building 3D models. In addition, although 3D models can be directly annotated, the process becomes challenging due to the low resolution of textures and the large size of the models. In this article, we propose a streamlined, open-access processing pipeline to reproject 2D image annotations onto 3D models using ray tracing. Using four underwater image datasets, we assessed the accuracy of annotation reprojection on 3D models and achieved successful georeferencing to centimetric accuracy. The combination of photogrammetric 3D models and accurate 2D annotations would allow the construction of a 3D representation of the landscape and could provide new insights into understanding species microdistribution and biotic interactions.

The fish disease model facilitates our understanding of disease dynamics, risk assessment for disease outbreaks, the response of the gut immune system, and the maintenance of ecosystem health. Here, we present a protocol for studying gut immunity modulation by infecting Lepidocephalichthys guntea, a loach fish, with Aeromonas hydrophila. We describe steps for performing intra-peritoneal injection on fish and a bath challenge. We detail procedures for conducting periodic population calculations during the infection phase to corroborate Aeromonas hydrophila invasion in the gut. For complete details on the use and execution of this protocol, please refer to Basak and Chakraborty.1.

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