Co-occurrence patterns of gut microbiome, antibiotic resistome and the perturbation of dietary uptake in captive giant pandas.

The microbiome is a key source of antibiotic resistance genes (ARGs), significantly influenced by diet, which highlights the interconnectedness between diet, gut microbiome, and ARGs. Currently, our understanding is limited on the co-occurrence among gut microbiome, antibiotic resistome in the captive giant panda and the perturbation of dietary uptake, especially for the composition and forms in dietary nutrition. Here, a qPCR array with 384 primer sets and 16 S rRNA gene amplicon sequencing were used to characterize the antibiotic resistome and microbiomes in panda feces, dietary bamboo, and soil around the habitat. Diet nutrients containing organic and mineral substances in soluble and insoluble forms were also quantified. Organic and mineral components in water-unextractable fractions were 7.5 to 139 and 637 to 8695 times higher than those in water-extractable portions in bamboo and feces, respectively, while the latter contributed more to the variation (67.5 %) of gut microbiota. Streptococcus, Prevotellaceae, and Bacteroides were the dominant genera in giant pandas. The ARG patterns in panda guts showed higher diversity in old individuals but higher abundance in young ones, driven directly by the bacterial community change and mobile genetic element mediation and indirectly by dietary intervention. Our results suggest that dietary nutrition mainly accounts for the shift of gut microbiota, while bacterial community and mobile genetic elements influenced the variation of gut antibiotic resistome.

Extrapolation of cytotoxic masked effects in planar in vitro assays.

The masking of specific effects in in vitro assays by cytotoxicity is a commonly known phenomenon. This may result in a partial or complete loss of effect signals. For common in vitro assays, approaches for identifying and quantifying cytotoxic masking are partly available. However, a quantification of cytotoxicity-affected signals is not possible. As an alternative, planar bioassays that combine high-performance thin layer chromatography with in vitro assays, such as the planar yeast estrogen screen (p-YES), might allow for a quantification of cytotoxically affected signals. Affected signals form a typical ring structure with a supressed or completely lacking centre that results in a double peak chromatogram. This study investigates whether these double peaks can be used for fitting a peak function to extrapolate the theoretical, unaffected signals. The precision of the modelling was evaluated for four individual peak functions, using 42 ideal, undistorted peaks from estrogenic model compounds in the p-YES. Modelled ED50-values from bisphenol A (BPA) experiments with cytotoxically disturbed signals were 13 times higher than for the apparent data without compensation for cytotoxicity (320 ± 63 ng versus 24 ± 17 ng). This finding has a high relevance for the modelling of mixture effects according to concentration addition that requires unaffected, complete dose-response relationships. Finally, we applied the approach to results of a p-YES assay on leachate samples of an elastomer material used in water engineering. In summary, the fitting approach enables the quantitative evaluation of cytotoxically affected signals in planar in vitro assays and also has applications for other fields of chemical analysis like distorted chromatography signals.

LandS: Vegetation modeling based on Ellenberg's ecological indicator values.

We present LandS, a new version of the Gras Model. The Gras Model was designed to simulate grassland development at local scales based on Ecological Indicator Values (EIVs) for different grassland management practices. In LandS, we complemented the existing set of EIVs with a second set representing several environmental factors: light, moisture, temperature, soil pH and nitrogen, also known as Ellenberg's EIVs. These new EIVs make the model more versatile and applicable to a wide range of sites across Central Europe. For example, it can be used on sites with dry or moist, acidic or calcareous soils in grassland or forest environments. We have also improved the implementation of the model by introducing version control and moving species and site-specific variables to data input files, so that species sets can be easily swapped for application in new study sites. We demonstrate the use and behavior of the model in two simulation experiments exploring interactions mediated by Ellenberg's EIVs, using input files to apply the model to different landscapes. We also provide detailed guidance on species selection and calibration, and discuss model limitations.•LandS is an improved version of the GraS Model for simulating vegetation development at the local scale.•It includes Ellenberg-like indicator values for environmental variables for inverse prediction of species occurrence and composition.•The model is now flexible enough to be used for study sites throughout Central Europe, using data input files for species initialization.

Conflicts of Interest in the Assessment of Chemicals, Waste, and Pollution.

Pollution by chemicals and waste impacts human and ecosystem health on regional, national, and global scales, resulting, together with climate change and biodiversity loss, in a triple planetary crisis. Consequently, in 2022, countries agreed to establish an intergovernmental science-policy panel (SPP) on chemicals, waste, and pollution prevention, complementary to the existing intergovernmental science-policy bodies on climate change and biodiversity. To ensure the SPP's success, it is imperative to protect it from conflicts of interest (COI). Here, we (i) define and review the implications of COI, and its relevance for the management of chemicals, waste, and pollution; (ii) summarize established tactics to manufacture doubt in favor of vested interests, i.e., to counter scientific evidence and/or to promote misleading narratives favorable to financial interests; and (iii) illustrate these with selected examples. This analysis leads to a review of arguments for and against chemical industry representation in the SPP's work. We further (iv) rebut an assertion voiced by some that the chemical industry should be directly involved in the panel's work because it possesses data on chemicals essential for the panel's activities. Finally, (v) we present steps that should be taken to prevent the detrimental impacts of COI in the work of the SPP. In particular, we propose to include an independent auditor's role in the SPP to ensure that participation and processes follow clear COI rules. Among others, the auditor should evaluate the content of the assessments produced to ensure unbiased representation of information that underpins the SPP's activities.

Monitoring metal patterns from urban and agrarian sites using the bumblebee Bombus terrestris as a bioindicator.

Honeybees are well-established bioindicators for different types of pollutants. This study aims to establish another species of the Apidae family as a bioindicator, with a distinct behaviour and life cycle. The bumblebee Bombus terrestris was used as a bioindicator for 12 metals. Bumblebee hives were placed at sampling sites in and around the city of Aachen, Germany, and metal concentrations were assessed using ICP-MS. Metal concentrations were compared to those found in honeybees described in the literature. Spatial differences in metal patterns were investigated by comparing two land-use types: urban and agrarian. Seasonal differences were compared by taking samples in spring and summer. All analysed metals were detected above the detection limit and within or even above the concentration range found in honeybees. Significant spatial differences were found for the metalloid B and the metal Cd with higher concentrations at the agrarian sites than the urban sites. Significant seasonal differences were found for 8 metals: Fe, Zn, Cu, Ni, Cd, and As concentrations were higher in summer than in spring, while B and V concentrations were higher in spring. To categorise the results, we applied the honeybee contamination index (HCI) and adapted it to bumblebee purposes. According to the HCI, only one agrarian site showed a high contamination level. This study shows that bumblebees are suitable bioindicators for metals. The obtained data can serve as a first baseline in the establishment of additional monitoring studies or risk assessments.

Environmental selenium volatilization is possibly conferred by promiscuous reactions of the sulfur metabolism.

Selenium deficiency affects many million people worldwide and volatilization of biogenically methylated selenium species to the atmosphere may limit Se entering the food chain. However, there is very little systematic data on volatilization at nanomolar concentrations prevalent in pristine natural environments. Pseudomonas tolaasii cultures efficiently methylated Se at these concentrations. Nearly perfect linear correlations between the spiked Se concentrations and Dimethylselenide, Dimethyldiselenide, Dimethylselenylsulfide and 2-hydroxy-3-(methylselanyl)propanoic acid were observed up to 80 nM. The efficiency of methylation increased linearly with increasing initial Se concentration, arguing that the enzymes involved are not constitutive, but methylation proceeds promiscuously via pathways of S methylation. From the ratio of all methylated Se and S species, one can conclude that between 0.30% and 3.48% of atoms were Se promiscuously methylated at such low concentrations. At concentrations higher than 640 nM (∼50 µg/L) a steep increase in methylation and volatilization was observed, which suggested the induction of specific enzymes. Promiscuous methylation at low environmental concentrations calls into question that view that methylated Se in the atmosphere is a result of a purposeful Se metabolism serving detoxification. Rather, the concentrations of methylated Se in the atmosphere may be "coincidental" i.e., determined by the activity of S cycling microorganisms. Further, a steep increase in methylation efficiency when surpassing a certain threshold concentration (here ∼50 µg/L) calls into question that natural methylation can be estimated from high Se spikes in laboratory systems, yet highlights the possibility of using bacterial methylation as an effective remediation strategy for media higher concentrated in Se.

Fermented biochar has a markedly different effect on fate of pesticides in soil than compost, straw, and a mixed biochar-product.

Current knowledge about how biochars affect the fate of pesticides in soil is based on studies that used pure biochars. After finding that an additional biological post-pyrolysis treatment, such as co-composting or lactic fermentation, is required for biochars for superior performance in temperate arable soils, a knowledge gap formed of how such further processed biochar products would affect the fate of pesticides in soil. This study compared the effects of a novel fermented biochar alone or mixed with biogas residues on the fate of two pesticides, 4-chloro-2-methylphenoxyacetic acid (MCPA) and metalaxyl-M, in a temperate arable soil to the traditional organic amendments wheat straw and compost. The fate of 14C-labeled MCPA was markedly affected in different ways. Fermented biochar effectively reduced the water-extractability and mineralization due to adsorption that was comparable to adsorption strengths reported for pure biochars. However, this effect was weak for the biochar mixed with biogas residues. Straw reduced water-extractable amounts due to increased biodegradation and formation of likely biogenic non-extractable residues of MCPA. In contrast, compost decelerated mineralization and increased the water solubility of the MCPA residues due to released dissolved organic matter. The amendments' effects were minor regarding 14C-metalaxyl-M, except for the fermented biochar which again reduced water-extractability and delayed degradation due to adsorption. Thus, the effects of the organic amendments differed for the two pesticide compounds with only the fermented biochar's effect being similar for both. However, this effect was no longer present in the mixed product containing 20% biochar. Our findings clearly show that biologically treated biochar-containing products can affect the fate of pesticides in soil very differently, also when compared to traditional organic amendments. Such impacts and their desirable and undesirable ecotoxicological implications need to be considered before the large-scale application of biochars to temperate arable soils.

Evaluating microbial contaminations of alternative heating oils.

Since 2008, European and German legislative initiatives for climate protection and reduced dependency on fossil resources led to the introduction of biofuels as CO2-reduced alternatives in the heating oil sector. In the case of biodiesel, customers were confronted with accelerated microbial contaminations during storage. Since then, other fuel alternatives, like hydrogenated vegetable oils (HVOs), gas-to-liquid (GtL) products, or oxymethylene ether (OME) have been developed. In this study, we use online monitoring of microbial CO2 production and the simulation of onset of microbial contamination to investigate the contamination potential of fuel alternatives during storage. As references, fossil heating oil of German refineries are used. Biodiesel blends with fossil heating oils confirmed the promotion of microbial activity. In stark contrast, OMEs have an antimicrobial effect. The paraffinic Fischer-Tropsch products and biogenic hydrogenation products demonstrate to be at least as resistant to microbial contamination as fossil heating oils despite allowing a diversity of representative microbes. Through mass spectrometry, elemental analysis, and microbial sequencing, we can discuss fuel properties that affect microbial contaminations. In summary, novel, non-fossil heating oils show clear differences in microbial resistance during long-term storage. Designing blends with an intrinsic resistance against microbial contamination and hence reduced activity might be an option.

Organic solvent free PbI2 recycling from perovskite solar cells using hot water.

Perovskite solar cells represent an emerging and highly promising renewable energy technology. However, the most efficient perovskite solar cells critically depend on the use of lead. This represents a possible environmental concern potentially limiting the technologies' commercialization. Here, we demonstrate a facile recycling process for PbI2, the most common lead-based precursor in perovskite absorber material. The process uses only hot water to effectively extract lead from synthetic precursor mixes, plastic- and glass-based perovskites (92.6 - 100% efficiency after two extractions). When the hot extractant is cooled, crystalline PbI2 in high purity (> 95.9%) precipitated with a high yield: from glass-based perovskites, the first cycle of extraction / precipitation was sufficient to recover 94.4 ± 5.6% of Pb, whereas a second cycle yielded another 10.0 ± 5.2% Pb, making the recovery quantitative. The solid extraction residue remaining is consequently deprived of metals and may thus be disposed as non-hazardous waste. Therefore, exploiting the highly temperature-dependent solubility of PbI2 in water provides a straightforward, easy to implement way to efficiently extract lead from PSC at the end-of-life and deposit the extraction residues in a cost-effective manner, mitigating the potential risk of lead leaching at the perovskites' end-of-life.

Transcriptomic and metabolomic changes in lettuce triggered by microplastics-stress.

Microplastics (MPs) are a global threat to the environment, and plant uptake of MP particles (≤0.2 µm) is a particular cause for concern. However, physiological and molecular mechanisms underlying MP-induced growth inhibition need to be clarified. Towards this goal, we conducted a hydroponic experiment to investigate the accumulation of MPs, changes in physiology, gene expression, and metabolites in lettuce from a series of concentrations of fluorescence-labelled polystyrene MPs (0, 10, 20, 30, 40, 50 mg L-1, ∼0.2 µm). Our results showed that MPs accumulated in the lettuce root tips and leaf veins, resulting in the hypertonic injury of lettuce, and the down-regulation of genes related to ion homeostasis. Stress-related genes were up-regulated, and sphingolipid metabolism increased in response to MP additions, causing increased biosynthesis of ascorbic acid, terpenoid, and flavonoids in root exudates. Our findings provide a molecular-scale perspective on the response of leafy vegetables to MP-stress at a range of concentrations. This enables more comprehensive evaluation of the risks of MPs to human health and the ecological environment.

Manure- and straw-derived biochars reduce the ecological risk of PBDE and promote nitrogen cycling by shaping microbiomes in PBDE-contaminated soil.

Pyrolysis of agricultural waste into biochar for soil remediation is a useful solid waste management strategy. However, it is still unclear how different agricultural feedstocks affect the properties of biochars and their effectiveness in remediation of PBDE-contaminated soil. In this study, we systematically investigated dynamic alterations of soil properties, microbial communities, and PBDE dissipation and bioavailability induced by the application of biochars from manure (MBC) and straw (SBC) to PBDE-contaminated soil. The results showed that soil properties, microbial community structure, and diversity changed differently with the incorporation of the two biochars. MBC had a larger surface area (17.4 m2/g) and a higher nutrient content (45.1% ash content), making it more suitable for use as a soil additive to improve soil quality and nutrient conditions, as well as to stimulate microbial growth. SBC showed higher adsorption capacity for 2,2',4,4'-Tetrabromodiphenyl Ether (BDE-47) (26.73 ± 0.65 mg/g), thus lowering the bioavailability and ecological risk of BDE-47 in soil. BDE-47 was stepwise debrominated into lower brominated PBDE by PBDE-degrading bacteria. MBC accelerated the debromination of BDE-47 (10.1%) by promoting PBDE-degrading microorganisms, while this was inhibited by SBC (3.5%) due to strong adsorption of BDE-47. In addition, we found that both types of biochar favored Nitrospirae bacteria and promoted N cycling. Overall, biochars from manure and straw can positively shape soil microbial communities differently by altering soil properties, soil fertility and nutrient availability, and the fate and the effects of contaminants, which ultimately led to a difference in the potential of biochars for their use in soil remediation.

Positively Charged Microplastics Induce Strong Lettuce Stress Responses from Physiological, Transcriptomic, and Metabolomic Perspectives.

Microplastics (MPs) can enter plants through the foliar pathway and are potential hazards to ecosystems and human health. However, studies related to the molecular mechanisms underlying the impact of foliar exposure to differently charged MPs to leafy vegetables are limited. Because the surfaces of MPs in the environment are often charged, we explored the uptake pathways, accumulation concentration of MPs, physiological responses, and molecular mechanisms of lettuce foliarly exposed to MPs carrying positive (MP+) and negative charges (MP-). MPs largely accumulated in the lettuce leaves, and stomatal uptake and cuticle entry could be the main pathways for MPs to get inside lettuce leaves. More MP+ entered lettuce leaves and induced physiological, transcriptomic, and metabolomic changes, including a decrease in biomass and photosynthetic pigments, an increase in reactive oxygen species and antioxidant activities, a differential expression of genes, and a change of metabolite profiles. In particular, MP+ caused the upregulation of circadian rhythm-related genes, and this may play a major role in the greater physiological toxicity of MP+ to lettuce, compared to MP-. These findings provide direct evidence that MPs can enter plant leaves following foliar exposure and a molecular-scale perspective on the response of leafy vegetables to differently charged MPs.

To be or not to be degraded: in defense of persistence assessment of chemicals.

Characterizing the degradation behavior of chemicals in the environment is a key component of chemical hazard and risk assessment. Persistence has been successfully characterized for readily and for slowly degradable chemicals using standardized tests, but for the third group of chemicals with intermediate degradability ("middle group"), the assessment is less straightforward. Whether chemicals of this group behave as persistent or not in a given environment depends on environmental factors such as the presence of sorbents that can limit the bioavailability of chemicals. Uncertainties associated with current persistence assessments of chemicals in the middle group do not imply that persistence assessment is generally inconsistent, too ambiguous for regulatory use, and not useful in chemical hazard and risk assessment. Given the complexity of the environmental factors influencing chemical degradation, and the diversity of commercial chemicals, it has to be accepted though that for chemicals in the middle group even improved testing methods will not remove all of the immanent heterogeneity in their persistence data. For cases with widely different but technically valid persistence data, a weight-of-evidence approach is necessary and the "benefit of the doubt" should follow the precautionary principle in order to protect human and ecosystem health. We maintain that technically valid persistence data, although they might be considered dissatisfying from a scientific point of view because of high variability or even inconclusiveness, can well be sufficient for regulatory purposes. As with anything, also in persistence assessment, the scientific logic aims for a mechanistic description of the processes involved, low uncertainty, and a comprehensive understanding derived from a broad empirical basis. If the scientific logic is used as a benchmark in the regulatory context, this may easily lead to "paralysis by analysis". While regulatory decisions should be based on sound science, discrepancies between scientific goals and regulatory needs and, consequently, different levels of requirements (must-have versus nice-to-have) for degradation studies need to be recognized and appreciated. We further advocate for enhancing consistency between regulatory persistence assessments ("one substance-one assessment"), which is currently not the case.

Freeze-thaw alternations accelerate plasticizers release and pose a risk for exposed organisms.

The application of plastic mulch films brings convenience to agricultural production, but also causes plastic waste that can be degraded into microplastics (MPs). However, little is known about the fate of plastic waste in agricultural ecosystem under freeze-thaw alternation in middle and high latitudes, as well as in highlands around the world. Whether the release of plasticizers, i.e. phthalate esters (PAEs), under such conditions would pose a potential risk to exposed organisms due to bioaccumulation is also unknown. To fill these data gaps, the agricultural fields in Liaoning of China with typical freeze-thaw alternation was selected as the study area. The transformation of plastic film was demonstrated by simulation freeze-thaw alternating from -30 to 20 â„ƒ. Soil samples were collected to investigate the patterns of MP composition, abundance, and distribution. Concurrently, the concentrations of two PAEs including bis(2-ethylhexyl) phthalate (DEHP) and diethyl phthalate (DEP) in soils were analyzed to provide information on the correlation between MPs abundance and PAEs concentrations as well as potential risks. The results showed that freeze-thaw alternating can accelerate the formation of MPs and release of PAEs from plastic waste. The abundance of MPs was positively correlated with the concentration of PAEs. Soil PAEs ranged from 3268 ± 213-6351 ± 110 µg/kg, indicating that over 40 % of the PAEs were transferred from plastic films to soils. Such residual amounts could pose risk for exposed organisms. Hence, the current study suggested that special concerns should be given to the release plasticizers in plastic waste of agricultural soils.

Rapid sequestration of perovskite solar cell-derived lead in soil.

Efficient and stable perovskite solar cells rely on the use of Pb species potentially challenging the technologies' commercialisation. In this study, the fate of Pb derived from two common perovskite precursors is compared to cationic lead in soil-water microcosm experiments under various biogeochemical conditions. The rapid and efficient removal of Pb from the aqueous phase is demonstrated by inductively coupled plasma mass spectrometry. Sequential soil extraction results reveal that a substantial amount of Pb is associated with immobile fractions, whereas a minor proportion of Pb may become available again in the long term, when oxygen is depleted (e.g. during water logging). X-ray absorption spectroscopy results reveal that the sorption of Pb on mineral phases represents the most likely sequestration mechanism. The obtained results suggest that the availability of leached Pb from perovskite solar cells is naturally limited in soils and that its adverse effects on soil biota are possibly negligible in oxic soils. All three Pb sources used behaved very similar in the experiments, wherefore we conclude that perovskite derived Pb will have a similar fate compared to cationic Pb, so that established risk assessment considerations for Pb remain legitimate.

Fate of 14C-labelled ionic organic chemicals in a water-sediment system and surface water.

The persistence assessment of organic chemicals is based on neutral reference substances. Therefore, our study aimed at investigating the influence of a chemical charge on the degradation of organic compounds in a water-sediment system (OECD 308) and surface water (OECD 309). We used radiolabelled 4-n-dodecylbenzenesulfonic acid sodium salt (14C-DS-, anionic), 4-n-dodecylbenzyltrimethylammonium chloride (14C-DA+, cationic) and 4-n-dodecylphenol (14C-DP, non-ionic) which are structurally similar except their charges. After 120 days of incubation in a water-sediment system, 68% (14C-DS-), 6% (14C-DA+) and 63% (14C-DP) of the applied radioactivity (AR) were mineralized. The formation of non-extractable residues (NER) after 120 days was highest for 14C-DA+ (33% AR), followed by 14C-DS- (19% AR) and 14C-DP (14% AR). Dissipation half-lives (DT50) at 12 °C decreased as follows: 14C-DA+ (346 days) ≫ 14C-DS- (47 days) > 14C-DP (30 days). After 60 days of incubation in surface water with suspended sediment, mineralization of 14C-DS-, 14C-DA+ and 14C-DP accounted for 63%, 7% and 58% AR, respectively. Highest NER formation was observed for 14C-DP (21% AR), followed by 14C-DA+ (14% AR) and 14C-DS- (9% AR). DT50 (12 °C) decreased as follows: 14C-DA+ (45 days) > 14C-DP (3 days) > 14C-DS- (2 days). We showed that a positive charge reduces the degradability of organic chemicals in both test systems. From a scientific point of view, simulation studies following OECD 309 should always be complimented by tests with high sorption capacity, e.g. OECD 308 and OECD 307 tests in order to assess the degradation of a compound, especially in case of cationic organic compounds.