Diverse Mixture Research Articles

Evidence of Oxidative Stress as a Mechanism of Pharmaceutical-Induced Toxicity in Amphibians

Amphibians, which are essential components of ecosystems, are susceptible to pharmaceutical contamination, a phenomenon of increasing concern owing to the widespread consumption and detection of pharmaceutical compounds in environmental matrices. This review investigates oxidative stress (OS) as the primary mechanism of drug toxicity in these organisms. The evidence gathered reveals that various pharmaceuticals, from antibiotics to anesthetics, induce OS by altering biomarkers of oxidative damage and antioxidant defense. These findings underscore the deleterious effects of pharmaceuticals on amphibian health and development and emphasize the necessity of incorporating OS biomarkers into ecotoxicological risk assessments. Although further studies on diverse amphibian species, drug mixtures, and field studies are required, OS biomarkers offer valuable tools for identifying sublethal risks. Furthermore, the development of more refined OS biomarkers will facilitate the early detection of adverse effects, which are crucial for protecting amphibians and their ecosystems. Ultimately, this review calls for continued research and mitigation strategies to safeguard biodiversity from pharmaceutical contamination.

A tuneable minimal cell membrane reveals that two lipid species suffice for life

All cells are encapsulated by a lipid membrane that facilitates their interactions with the environment. How cells manage diverse mixtures of lipids, which dictate membrane property and function, is experimentally challenging to address. Here, we present an approach to tune and minimize membrane lipid composition in the bacterium Mycoplasma mycoides and its derived ‘minimal cell’ (JCVI-Syn3A), revealing that a two-component lipidome can support life. Systematic reintroduction of phospholipids with different features demonstrates that acyl chain diversity is more important for growth than head group diversity. By tuning lipid chirality, we explore the lipid divide between Archaea and the rest of life, showing that ancestral lipidomes could have been heterochiral. However, in these simple organisms, heterochirality leads to impaired cellular fitness. Thus, our approach offers a tunable minimal membrane system to explore the fundamental lipidomic requirements for life, thereby extending the concept of minimal life from the genome to the lipidome.

Evaluative criteria for least-cost economic analysis of citywide inclusive sanitation: a scoping review

ABSTRACT Compared to health interventions, where evaluative criteria have been established, uncertainty about how comparative economic analysis methodologies should be applied to heterogeneous urban sanitation interventions in cities of low- and middle-income countries persists. Gaps between least-cost principles and current approaches constrain emerging citywide inclusive sanitation (CWIS) planning frameworks. Furthermore, a lack of comparable evidence about the economic efficiency of service provision interventions impacts the delivery of urban sanitation as an inclusive public service. This review scopes how least-cost principles may be applied to methodologies for urban sanitation interventions in two iterations. We devise and discuss evaluative criteria, embodying CWIS principles, that will improve the quality of these methodologies. They include how (i) the inclusion of diverse cost perspectives, particularly populations with marginal access to sanitation, are accommodated within an analysis; (ii) a context-specific without-project case may be used as an appropriate basis for comparison; (iii) tangible and intangible costs and outcomes may influence decisions; and (iv) diverse mixtures of interventions at different scales may be compared with integrity. The resulting evaluative criteria define principles that may inform future systematic reviews on practices that enhance CWIS planning frameworks.

A tuneable minimal cell membrane reveals that two lipid species suffice for life.

All cells are encapsulated by a lipid membrane which facilitates the interaction between life and its environment. How life exploits the diverse mixtures of lipids that dictate membrane property and function has been experimentally challenging to address. We introduce an approach to tune and minimize lipidomes in Mycoplasma mycoides and the Minimal Cell (JCVI-Syn3A) revealing that a 2-component lipidome can support life. Systematically reintroducing phospholipid features demonstrated that acyl chain diversity is more critical for growth than head group diversity. By tuning lipid chirality, we explored the lipid divide between Archaea and the rest of life, showing that ancestral lipidomes could have been heterochiral. Our approach offers a tunable minimal membrane system to explore the fundamental lipidomic requirements for life, thereby extending the concept of minimal life from the genome to the lipidome.

Dissolved organic matter (DOM) rather than warming and eutrophication directly affects partial pressure of CO2 (pCO2) in mesocosm systems

Environmental warming and eutrophication pose significant challenges to shallow lake systems, where dissolved organic matter (DOM) serves as a diverse and intricate mixture of organic macromolecules, playing a pivotal role in aquatic ecosystems. Despite its complexity, comprehending the interplay between environmental changes and DOM composition alterations and their subsequent impacts on aqueous CO2 partial pressure (pCO2) is essential for a better understanding of carbon cycling. Yet, our current understanding in this realm remains limited. To address this gap, mesocosm systems were established to investigate how elevated water temperature and eutrophication, alongside changes in DOM composition, influence pCO2 dynamics. Results indicate that while temperature and nutrient levels do not directly influence pCO2 fluctuations, they indirectly affect aqueous pCO2 through their modulation of DOM composition. Elevated temperature and nutrient concentrations notably enhance both the production and degradation of indigenous protein-like organic matter and increase the accumulation of humic-like organic compounds, with phosphorus released from sediment playing a particularly significant role. Furthermore, the degradation rate of protein-like organic matter significantly exceeds its accumulation rate. On the other hand, the impact of water eutrophication on DOM composition surpasses that of temporal temperature variations, with a 2∼4 °C temperature rise showing minimal effects on DOM composition. Notably, the degradation of protein-like organic matter markedly increases aqueous pCO2, while the rise in humic-like organic matter in water exerts minimal influence on pCO2 concentrations. A comprehensive understanding of carbon cycling processes under environmental changes will facilitate effective management of lake ecosystems and the advancement of carbon mitigation technologies.

Recent advances in capillary electrophoresis-mass spectrometry analysis of ‍trace biomolecules

In recent years, various trace bioanalysis methods have been developed, including single-cell transcriptome analysis methods. As the sample volume and amount of biomolecules contained therein are extremely limited, development of new single-cell analysis methods require extremely high-level techniques. It is necessary to design an appropriate analysis system that integrates a highly sensitive detection system and a pretreatment protocol for minimizing sample loss, where separation method is especially important for analyzing diverse mixtures of biomolecules. Among them, capillary electrophoresis (CE) can separate biomolecules in nanoliter-scale solutions with high resolution, making it highly compatible with trace samples such as single cells. By combining with highly sensitive nano-electrospray ionization-mass spectrometry (MS), it is possible to detect nanomolar to sub-nanomolar biomolecules, which can be further improved by using online sample preconcentration methods. These highly sensitive analytical techniques have made it possible to analyze trace amounts of metabolites, proteins, lipids, etc. This review paper summarizes the research on CE-MS trace bioanalysis that has been reported to date, with a focus on single-cell analysis.

Wavelength-Selective Xanthene-Based Monochromophoric Photoremovable Protecting Groups for Tuning Soft Matter Material Properties.

Photocontrolled deprotection of specific functional groups has garnered significant interest over the past two decades. Notably, the selective deprotection of distinct groups based on wavelength has emerged as a prominent focus in recent research. The achievement of this objective has primarily involved the utilization of linker-based bichromophoric systems and diverse cocktail mixtures of photoresponsive protecting groups (PRPGs), each responsive to varying wavelengths of light. Herein, we present the first wavelength-selective monochromophoric system based on a hydroxanthene moiety, enabling the wavelength-selective release of two distinct functionalities under 450 and 600 nm light, respectively. The mechanism of the wavelength-selective photodegradation was thoroughly investigated by 1H NMR, UV-vis, and fluorescence spectroscopy, suggesting a proton-coupled electron transfer mechanism in the first photorelease step and electron transfer based arylmethyl type of photorelease in the second step. The utility of the xanthene-based wavelength-selective PRPGs was demonstrated in the multistep degradation of microparticles and dual-color tuning of polymer chain architecture, thus opening an avenue to design advanced photoreactive wavelength-controlled systems for applications in soft matter materials.

A brief history and future directions of dielectrophoretic filtration: A review.

Dielectrophoresis (DEP) is an electrokinetic effect first studied in the early 20th century. Since then, DEP has gained significant interest in research, owing to its ability to solve particle separation problems in various industries. Dielectrophoretic filtration (DEP filtration) is a separation method using DEP to filter a wide range of microparticles, from bacterial cells to catalytic particles. DEP filtration can selectively separate particles based on size or dielectric properties, recover trapped particles and avoid common problems associated with mechanical filtration based on pore size (e.g. pressure drops and regular filter replacements). This review describes the simple beginnings of DEP filtration and how our understanding and applications for DEP filtration have progressed over time. A brief section of DEP theory as well as a note on the general outlook for DEP filtration in the future is presented. DEP filtration offers an exciting opportunity to selectively separate diverse particle mixtures. To achieve such a feat, technical challenges such as Joule Heating and low throughputs must be addressed.

Dispersal and connectivity modelling simulations for invertebrate larvae passing through the Strait of Gibraltar

Abstract The link between the northeastern Atlantic and Mediterranean Sea created by the Strait of Gibraltar and the adjacent Iberian and Moroccan coasts marks remarkable transition areas between distinct environments that harbour a diverse mixture of species. The area is interesting regarding marine connectivity and the transport of pelagic invertebrate larvae, crucial knowledge to manage over-exploited populations, and minimize the impacts of climate change and anthropogenic activities. Biophysical models were developed, combining oceanic and particle-tracking Lagrangian simulations with in situ zooplankton distribution data. The conditions driving the larval exchange between the sub-basins and the connectivity throughout the region were explored, using crustacean decapod larvae as biological references. The potential exchange between both sub-basins was confirmed, although specific larval traits revealed contrasting scenarios. The simulations showed that slope-dwelling and mesopelagic larvae have advantage when crossing from the Alboran into the Atlantic, in comparison with shelf-dwelling and epipelagic larvae. Transport pathways and retention areas were identified, and passive drifts were shown to increase the dispersal range of the simulations. The spatial origin of the larval release, larval duration, vertical distribution, and the interaction of larvae with the oceanic features are presented as the main factors impacting the effective larval input into the Atlantic or Mediterranean basins.

The thawing process in gentamicin and polyethylene glycol hydrogel: The impact of selected parameters on the calculation of activation energy using the methods of Belichmaer and Duswalt

AbstractThe subject of the research conducted by differential scanning calorimetry (DSC) was hydrated mixtures of polyethylene glycol and gentamicin (GEN) in a 1:1 (w/w) ratio. In this way, a system with a particularly high ability to form hydrogen bonds was created. It was postulated that studying the effective activation energy (EA) of the thawing process of diverse hydrated polymer mixtures will indirectly allow for an attempt to assess the distribution of water molecules in the examined material in future studies. This article presents a detailed analysis of the impact of selected parameters on the calculation of EA using the Belichmaer and Duswalt methods. The research focused on assessing how the interpolation method affects the accuracy of results obtained from these techniques. The interpolation method was analyzed in terms of its ability to accurately replicate experimental data. By evaluating the impact of the number of points from the DSC curve used in linear interpolation, a standard deviation from the average value of EA was obtained, respectively: 2.4% for the Belichmaer method and 1.9% for the Duswalt method. The article also shows that by using both methods simultaneously, it is possible to detect the moment when additional endothermic and exothermic reaction activity becomes present in the reaction curve. This allowed for the avoidance of the depletion error. Computer modeling and simulations were used to compare the effectiveness of both techniques in various scenarios. The results showed that the choice of interpolation method and taking into account the depletion error significantly affect the accuracy of calculating EA. The computed activation energy via Duswalt method was EA = 363.32 kJ mol−1 with a standard deviation of 7.07 kJ mol−1 and using the Belichmaer method the EA was 364.81 kJ mol−1, with a standard deviation of 9.10 kJ mol−1.

Phylogenomics and biogeography of sawflies and woodwasps (Hymenoptera, Symphyta)

Phylogenomic approaches have recently helped elucidate various insect relationships, but large-scale comprehensive analyses on relationships within sawflies and woodwasps are still lacking. Here, we infer the relationships and long-term biogeographic history of these hymenopteran groups using a large dataset of 354 UCE loci collected from 385 species that represent all major lineages. Early Hymenoptera started diversifying during the Early Triassic ∼249 Ma and spread all over the ancient supercontinent Pangaea. We recovered Xyeloidea as a monophyletic sister group to other Hymenoptera and Pamphilioidea as sister to Unicalcarida. Within the diverse family Tenthredinidae, our taxonomically and geographically expanded taxon sampling highlights the non-monophyly of several traditionally defined subfamilies. In addition, the recent removal of Athalia and related genera from the Tenthredinidae into the separate family Athaliidae is supported. The deep historical biogeography of the group is characterised by independent dispersals and re-colonisations between the northern (Laurasia) and southern (Gondwana) palaeocontinents. The breakup of these landmasses led to ancient vicariance in several Gondwanan lineages, while interchange across the Northern Hemisphere has continued until the Recent. The little-studied African sawfly fauna is likewise a diverse mixture of groups with varying routes of colonization. Our results reveal interesting parallels in the evolution and biogeography of early hymenopterans and other ancient insect groups.

P-802 Exploring the effects of phthalate mixtures on human endometrial organoids: Insights into transcriptomic and proteomic profiles throughout the menstrual cycle

Abstract Study question Does the exposure of human endometrial organoids (hEOs) to phthalates influence their transcriptomic and proteomic profiles, particularly during the window of implantation? Summary answer The mixture of phthalates altered the expression of several genes which are essential during the window of implantation. What is known already hEOs display a robust responsiveness to steroid hormones and accurately reproduce essential morphological features observed in the receptive endometrium in vivo. The conventional approach to evaluating chemical health risks has predominantly been centered on the assessment of individual substances, overlooking the reality that individuals are regularly exposed to diverse mixtures of phthalates, a class of Endocrine Disrupting Chemicals (EDCs). There is a limited number of studies indicating that phthalates and their combinations can have a negative impact on the endometrium. To this purpose, we explore the effect of this mixture on endometrial receptivity during the Window Of Implantation (WOI). Study design, size, duration Endometrial tissues (n = 6) used for this study were obtained, with written informed consent from all participants. The endometrial organoids were generated from the biopsies and treated with hormones to mimic the WOI. Samples were then exposed with a mixture of phthalates in order to study the transcriptomic and proteomic changes on these cells throughout the menstrual cycle, especially the WOI. Participants/materials, setting, methods hEOs were generated from endometrial biopsies of healthy volunteers (n = 6). Organoids were cultured for 4 days and then divided into: untreated without any hormonal treatment; stimulated with 10 nM estrogen (E2); treated with 10 nM estrogen, 1μM progesterone (P4) and 1μM cyclic adenosine monophosphate (cAMP). EOs were further on exposed to the phthalate mixture. Proteomic profiling was performed using 2-D electrophoresis and Mass spectrometry. Samples were also subject to transcriptomic analysis, by RNA sequencing. Main results and the role of chance hEOs present a suitable in vitro model for the screening of the impact of phthalate mixtures on the endometrium throughout the menstrual cycle. Cytotoxicity test confirmed that the chosen concentration (100x of the value referring to the geometrical means in urine) of the mixture is not toxic for the cells. The transcriptomic and proteomic analyses revealed several up-regulated or down-regulated genes which are involved into pivotal pathways implicated to the WOI. Limitations, reasons for caution This study represents a preliminary experiment. All results need to be validated on an enlarged number of samples to provide more statistical strength. Wider implications of the findings hEOs offer a robust in vitro model to simulate and gain a more profound comprehension of the consequences arising from the combinatory effects of EDCs, on the endometrium. This model proves valuable in examining the impact of various EDCs across different phases of the menstrual cycle. Trial registration number not applicable

Impacts of polycultural cropping on crop yields and biodiversity: A systematic map protocol

Abstract Agriculture is currently the largest driver of biodiversity‐loss worldwide. There is a critical need to develop agricultural systems that protect and promote biodiversity, while also meeting local and global food needs. Ecological theory suggests that cultivating crops in diverse mixtures both maximises niche occupancy and generates additional niches, generating both higher yields and higher biodiversity than cultivation in monocultures. A large and growing body of agronomic research provides strong evidence for the potential productivity and biodiversity benefits of in‐field crop‐diversification strategies. This protocol sets out a methodology for a systematic map of evidence on yield‐ and biodiversity‐outcomes in ‘polycultures’—systems of cultivation where multiple food crop species are grown together simultaneously at field‐scale. Systematic searches with a global scope will be conducted in Web of Science Core Collection, Scopus, and CAB Abstracts to yield a comprehensive sample of relevant published, peer‐reviewed literature. The articles returned by searches will be subject to eligibility screening according to pre‐defined inclusion criteria, at successive stages (title and abstract, and full text). Following this, we will extract data from eligible studies on study designs, survey and sampling protocols, experimental treatments and comparators, and outcomes measured. A narrative synthesis, illustrated by tables, figures and maps, will describe the quantity and characteristics of the available evidence on the impacts of polycultural cropping on (i) crop yields, and (ii) agroecosystem biodiversity. We will also identify the research that examines these two outcomes in parallel. The synthesis will be accompanied by a published database containing bibliographic information as well as data (see ‘2’) on study characteristics and outcomes. Practical implications. The anticipated systematic map will provide a synthesis of current evidence regarding the potential yield‐ and biodiversity‐impacts of polycultural cropping. We will identify key knowledge gaps, and ‘clusters’ of related evidence that could lend themselves to further quantitative synthesis via subsequent systematic reviews and/or meta‐analyses. As such, the work will facilitate future appraisal of the potential of polyculture as a tool to bridge the currently conflicting priorities of food production on the one hand and biodiversity conservation on the other.

Topology of Pulsar Profiles (ToPP)

Some of the most important information on a radio pulsar is derived from its average pulse profile. Many early pulsar studies were necessarily based on only a few such profiles. In these studies, discrete profile components were linked to emission mechanism models for individual stars through human interpretation. For the population as a whole, profile morphology must reflect the geometry and overall evolution of the radio emitting regions. The problem, however, is that this population is becoming too large for individual intensive studies of each source. Moreover, connecting profiles from a large collection of pulsars rapidly becomes cumbersome. In this article, we present ToPP, the first-ever unsupervised method to sort pulsars by profile-shape similarity using graph topology. We applied ToPP to the publicly available European Pulsar Network profile database, providing the first organised visual overview of multi-frequency profiles representing 90 individual pulsars. We found discrete evolutionary tracks varying from simple single-component profiles at all frequencies towards diverse mixtures of more complex profiles with frequency evolution. The profile evolution is continuous, extending out to millisecond pulsars, and does not fall into sharp classes. We interpret the profiles as being a mixture of pulsar core-cone emission type, spin-down energetics, and the line-of-sight impact angle towards the magnetic axis. We show how ToPP can systematically classify sources into the Rankin empirical profile scheme. ToPP comprises one of the key unsupervised methods that will be essential to exploring upcoming pulsar census data, such as the data expected from the Square Kilometer Array.

An experimental platform for semi-autonomous kinetic model refinement combining optimal experimental design, computer-controlled experiments, and optimization leads to new understanding of N[formula omitted]O + O

Automated platforms could enable the unprecedented pace required for modeling the countless proposed alternative fuels relevant to future technologies, but existing platforms rely exclusively on automatically generated computational data and lack experimental validation. Here, we introduce an experimental platform for rapid kinetic model refinement that combines optimal experimental design, computer-controlled experiments, and optimization—each of which are tailored in novel ways to form a cohesive platform together. The optimal experimental design considers (1) realistic uncertainties in both experimental conditions and measurements and (2) diverse reactant mixtures that can include “chemical sensitizers,” which are not necessarily reactants in a specified Quantity of Interest (QoI) but sensitize kinetic information relevant to a QoI. Similarly, our High-Throughput Jet-Stirred Reactor (HT-JSR) features (1) rapid multi-species diagnostics that can measure dozens of species within minutes, (2) a flow delivery system that can prepare up to ∼10-component reactant mixtures, and (3) computer-controlled operation of all components. Finally, a post-processing code automatically retrieves data from the instruments, quantifies uncertainties in both experimental conditions and measurements, and produces self-contained files usable for optimization in our MultiScale Informatics software. This platform is demonstrated for the rate constant for N2O + O ⇌ N2 + O2 as the QoI where, unlike for N2O + O ⇌ NO + NO, proposed values at ∼1000 K span ∼5 orders of magnitude yet give equally good agreement with previous experimental data—suggesting that previous experiments fail to constrain the branching ratio of N2O + O. The results show that optimal conditions with more species as both reactants and analytes—particularly NO2 as both a reactant and analyte—enable unambiguous discrimination of the main products of N2O + O. Specifically, the data preclude N2 + O2 as the main products at ∼1000 K—contrary to most recently proposed values.Novelty and significance statementWe introduce a novel experimental platform for rapid kinetic model refinement that combines optimal design, computer-controlled experiments, and optimization—each uniquely tailored to form a cohesive platform. It notably employs high levels of automation, high-throughput multi-species diagnostics, and uniquely diverse reactant mixtures to accelerate scientific discovery. This platform is shown to achieve a goal that no previous experiment has: decipher the main products of N2O + O. This success—attributable to the platform’s unique design principles—demonstrates the effectiveness of this experimental platform as a tool for accelerating scientific discovery and kinetic model development.

Novel Anti-Enterovirus A71 Compounds Discovered by Repositioning Antivirals from the Open-Source MMV Pandemic Response Box.

The open-source drug library, namely, MMV Pandemic Response Box, contains 153 antiviral agents, a chemically and pharmacologically diverse mixture of early-stage, emerging anti-infective scaffolds, and mature compounds currently undergoing clinical development. Hence, the Pandemic Response Box might contain compounds that bind and interfere with target molecules or cellular pathways that are conserved or shared among the closely related viruses with enterovirus A71 (EV-A71). This study aimed to screen antiviral agents included in the Pandemic Response Box for repurposing to anti-EV-A71 activity and investigate the inhibitory effects of the compounds on viral replication. The compounds' cytotoxicity and ability to rescue infected cells were determined by % cell survival using an SRB assay. The hit compounds were verified for anti-EV-A71 activity by virus reduction assays for viral RNA copy numbers, viral protein synthesis, and mature particle production using qRT-PCR, Western blot analysis, and CCID50 assay, respectively. It was found that some of the hit compounds could reduce EV-A71 genome replication and protein synthesis. D-D7 (2-pyridone-containing human rhinovirus 3C protease inhibitor) exhibited the highest anti-EV-A71 activity. Even though D-D7 has been originally indicated as a polyprotein processing inhibitor of human rhinovirus 3C protease, it could be repurposed as an anti-EV-A71 agent.

Pollutant removal efficiency of bare and planted rain gardens with diverse planting mixtures.

This study examines the influence of planting mixture variations on the quality of the percolated water of the rain garden with and without plants. Six planting mixtures in experimental rain gardens have been used. It has been noted that pollutant removal efficiency of RG can exhibit variations based on specific parameters. Notably, RG6, utilizing a planting mix of 75% topsoil and 25% compost, demonstrated the highest performance. These results draw attention to the critical role of the specific planting mixtures in influencing the performance of vital parameters related to pollutant removal. The observation shows that RG5 exhibits exceptional removal efficiency in pH, Total Suspended Solids (TSS), Biological Oxygen Demand (BOD), and Chemical Oxygen Demand (COD), and RG6 performs best in electrical conductivity (EC), Total Dissolved Solids (TDS), Total Nitrogen (TN), and Total Phosphorus (TP) removal. In particular, when analyzing pollutant removal on a surface with Madagascar periwinkle plants, RG6 emerges as the most effective, achieving an impressive efficiency of approximately 49%. For the bare surface, pollutant removal efficiency is 40%. The study outcome will be useful in deciding the composition of the planting mixture, which will keep the rain garden to improve quality and quantitatively hydrological performance, lowering urban flooding magnitude.

Friction Stir Welding: A Comprehensive Review of Non-Metallic Particle Reinforcement in Joints

Friction Stir Welding (FSW), a solid-state joining technique, has demonstrated superior efficiency in welding metal-matrix-reinforced composite joints. FSW has garnered significant attention in recent years as a viable technique for joining similar and dissimilar materials, particularly those reinforced with non-metallic particles. By utilizing diverse mixture of reinforcement particles and base matrices, FSW outperforms traditional fusion joining methods in terms of effectiveness and reliability. Despite significant progress, challenges persist in achieving a homogeneous distribution of Non-metallic particles in the weld zone, which directly impacts macrostructural and microstructural characteristics. Moreover, the mechanical properties of these welds are intricately linked to process parameters, influencing grain enhancement and reinforcement particle distribution. This review critically evaluates these aspects, providing insights into the current understanding and highlighting areas for future research.

Tracking SARS-CoV-2 variants of concern in wastewater: an assessment of nine computational tools using simulated genomic data.

Wastewater-based surveillance (WBS) is an important epidemiological and public health tool for tracking pathogens across the scale of a building, neighbourhood, city, or region. WBS gained widespread adoption globally during the SARS-CoV-2 pandemic for estimating community infection levels by qPCR. Sequencing pathogen genes or genomes from wastewater adds information about pathogen genetic diversity, which can be used to identify viral lineages (including variants of concern) that are circulating in a local population. Capturing the genetic diversity by WBS sequencing is not trivial, as wastewater samples often contain a diverse mixture of viral lineages with real mutations and sequencing errors, which must be deconvoluted computationally from short sequencing reads. In this study we assess nine different computational tools that have recently been developed to address this challenge. We simulated 100 wastewater sequence samples consisting of SARS-CoV-2 BA.1, BA.2, and Delta lineages, in various mixtures, as well as a Delta-Omicron recombinant and a synthetic 'novel' lineage. Most tools performed well in identifying the true lineages present and estimating their relative abundances and were generally robust to variation in sequencing depth and read length. While many tools identified lineages present down to 1 % frequency, results were more reliable above a 5 % threshold. The presence of an unknown synthetic lineage, which represents an unclassified SARS-CoV-2 lineage, increases the error in relative abundance estimates of other lineages, but the magnitude of this effect was small for most tools. The tools also varied in how they labelled novel synthetic lineages and recombinants. While our simulated dataset represents just one of many possible use cases for these methods, we hope it helps users understand potential sources of error or bias in wastewater sequencing analysis and to appreciate the commonalities and differences across methods.

Untargeted Metabolomics Reveals Fruit Secondary Metabolites Alter Bat Nutrient Absorption

The ecological interaction between fleshy fruits and frugivores is influenced by diverse mixtures of secondary metabolites that naturally occur in the fruit pulp. Although some fruit secondary metabolites have a primary role in defending the pulp against antagonistic frugivores, these metabolites also potentially affect mutualistic interactions. The physiological impact of these secondary metabolites on mutualistic frugivores remains largely unexplored. Using a mutualistic fruit bat (Carollia perspicillata), we showed that ingesting four secondary metabolites commonly found in plant tissues affects bat foraging behavior and induces changes in the fecal metabolome. Our behavioral trials showed that the metabolites tested typically deter bats. Our metabolomic surveys suggest that secondary metabolites alter, either by increasing or decreasing, the absorption of essential macronutrients. These behavioral and physiological effects vary based on the specific identity and concentration of the metabolite tested. Our results also suggest that a portion of the secondary metabolites consumed is excreted by the bat intact or slightly modified. By identifying key shifts in the fecal metabolome of a mutualistic frugivore caused by secondary metabolite consumption, this study improves our understanding of the effects of fruit chemistry on frugivore physiology.