Ecotoxicological assessment of sanitary sewer overflows and rainfall dynamics offers insights into conditions for potential adverse ecological outcomes.

Sewer overflows are an environmental concern due to their potential to introduce contaminants that can adversely affect downstream aquatic ecosystems. As these overflows can occur during rainfall events, the influence of rainwater ingress from inflow and infiltration on raw untreated wastewater (influent) within the sewer is a critical factor influencing the dilution and toxicity of the contaminants. The Vineyard sewer carrier in the greater city of Sydney, Australia, was selected for an ecotoxicological investigation of a sanitary (separate from stormwater) sewerage system and a wet-weather overflow (WWO). Three influent samples were collected representing dry-weather (DW), intermediate wet-weather (IWW) and wet-weather (WW). In addition, a receiving water sample was also collected downstream in Vineyard Creek (WW-DS) coinciding with a WWO. We employed direct toxicity assessment (DTA) and toxicity identification evaluation (TIE) approaches to gain comprehensive insights into the nature and magnitude of the impact on influent from rainwater ingress into the sewer. Three standard ecotoxicological model species, a microalga, Chlorella vulgaris, the water flea, Ceriodaphnia dubia and the midge larva, Chironomus tepperi were used for both acute and chronic tests. The study revealed variable toxicity responses, with the sample of influent collected in wet-weather displaying lower toxicity compared to the dry-weather sample of influent. Ammonia, and metals, were identified in dry weather as contributors to the observed toxicity, however, this risk was alleviated through rainwater ingress in wet-weather with further dilution within the receiving water. Based on toxicity data, dilutions of influent to minimise effects on C. vulgaris and C. dubia ranged from 1 in 12 in DW to 1 in 2.8 in WW, and further diminished in the receiving water to 1 in 1.8. The successful application of ecotoxicological approaches enabled the assessment of cumulative effects of contaminants in influent, offering valuable insights into the sanitary sewer system under rainwater ingress.

Transcriptomic Point of Departure (tPOD) of androstenedione in zebrafish embryos as a potential surrogate for chronic endpoints.

The transcriptomic Point of Departure (tPOD) is increasingly used in ecotoxicology to derive quantitative endpoints from RNA sequencing studies. Utilizing transcriptomic data in zebrafish embryos as a New Approach Methodology (NAM) is beneficial due to its acknowledgment as an alternative to animal testing under EU Directive 2010/63/EU. Transcriptomic profiles are available in zebrafish for various modes of action (MoA). The limited literature available suggest that tPOD values from Fish Embryo Toxicity (FET) tests align with, but are generally lower than, No Observed Effect Concentrations (NOEC) from long-term chronic fish toxicity tests. In studies with the androgenic hormone androstenedione in a Fish Sexual Development Test (FSDT), a significant shift in the sex ratio towards males was noted at all test concentrations, making it impossible to determine a NOEC (NOEC <4.34 µg/L). To avoid additional animal testing in a repetition of the FSDT and adhere to the 3Rs principle (replacement, reduction, and refinement), a modified zebrafish FET (zFET) was conducted aiming to determine a regulatory acceptable effect threshold. This involved lower concentration ranges (20 to 6105 ng/L), overlapping with the masculinization-observed concentrations in the FSDT. The tPOD analysis in zFET showed consistent results with previous FSDT findings, observing strong expression changes in androgen-dependent genes at higher concentrations but not at lower ones, demonstrating a concentration-response relationship. The tPOD values for androstenedione were determined as 24 ng/L (10th percentile), 60 ng/L (20th gene), and 69 ng/L (1st peak). The 10th percentile tPOD value in zFET was 200 times lower than the lowest concentration in the FSDT. Comparing the tPOD values to literature suggests their potential to inform on the NOEC range in FSDT tests.

Ecotoxicological effects of triclosan on Lemna minor: bioconcentration, growth inhibition and oxidative stress.

Triclosan (TCS), an emerging pollutant, is a notable contributor to adverse impacts on aquatic organisms due to its widespread use during COVID-19 and hydrophobic properties. There is extensive documented literature on TCS toxicity in commercially important fish species; however, studies on aquatic plants remain limited. In this prelude, the present study aims to evaluate the effect of TCS on Lemna minor, a commercially important aquatic plant species for 7 days. The results showed dose-dependent significant alterations in growth, pigments and stress enzymes of L. minor at varied concentrations of TCS (1 to 8 mg L-1). Median inhibitory concentration (IC50) was found to be 4.813 mg L-1. Total chlorophyll and carotenoid levels decreased 73.11 and 81.83%, respectively after 7 days of TCS exposure. A significant increase in catalase and superoxide dismutase activity was observed in TCS exposed groups as compared to the control. Bioconcentration factor was found to be in the range of 5.855 to 37.129 signifying TCS ability to accumulate and transfer through the food chain. Scanning electron microscopy (SEM) analysis showed deformation in the cell surface and alteration of stroma morphology of TCS exposed groups. Furthermore, the Fourier transform infrared spectroscopy (FTIR) study also revealed that higher concentrations of TCS could cause alteration in the functional groups in the plant. This study demonstrates that TCS negatively impacts the growth and metabolism of primary producers, offering crucial insights into its interactions with aquatic plants and establishing baseline information essential for crafting effective mitigation strategies for TCS contamination in aquatic environments.

Ecotoxicological study of seven pharmaceutically active compounds: Mixture effects and environmental risk assessment.

Pharmaceutically active compounds (PhACs) have been detected in several aquatic compartments, which has been of environmental concern since PhACs can cause adverse effects on the aquatic ecosystem at low concentrations. Despite the variety of PhACs detected in surface water, ecotoxicological studies are non-existent for many of them, mainly regarding their mixture. In addition, water bodies can continuously receive the discharge of raw or treated wastewater with micropollutants. Thus, PhACs are subject to mixture and interactions, potentiating or reducing their toxicity. Therefore, the present study evaluated the toxicity on Aliivibrio fischeri of seven PhACs, which still needs to be explored in the literature. The effects were evaluated for the PhACs individually and for their binary and tertiary mixture. Also, the experimental effects were compared with the concentration addition (CA) and independent action (IA) models. Finally, an environmental risk assessment was carried out. Fenofibrate (FEN), loratadine (LOR), and ketoprofen (KET) were the most toxic, with EC50 of 0.32 mg L-1, 6.15 mg L-1 and 36.8 mg L-1, respectively. Synergistic effects were observed for FEN + LOR, KET + LOR, and KET + FEN + LOR, showing that the CA and IA may underestimate the toxicity. Environmental risks for KET concerning algae, and LOR e 17α-ethynylestradiol (EE2) for crustaceans and fish were high for several locations. Besides, high removals by wastewater treatment technologies are required to achieve the concentrations necessary for reducing KET and LOR risk quotients. Thus, this study contributed to a better understanding of the toxic interactions and environmental risks of PhACs.

Metal-based nanomaterials in aquatic environments: What do we know so far about their ecotoxicity?

The wide range of applications of nanomaterials (NM) in different fields has led to both uncontrolled production and release into environmental compartments, such as aquatic systems, where final disposal occurs. Some efforts have been made to estimate their concentrations in environmental matrices; however, little is known about the actual effects of environmental NM concentrations on biota. The aims of the present review are to (i) expose the state of the art of the most applied NM and their actual concentrations regarding how much is being released to the aquatic environment and which are the predicted ones; (ii) analyze the current literature to elucidate if the aforementioned conditions were proven to cause deleterious effects on the associated organisms; and (iii) identify gaps in the knowledge regarding whether the actual NM concentrations are harmful to aquatic biota. These novel materials are expected to being released into the environment in the range of hundreds to thousands of tons per year, with Si- and Ti-based NM being the two most important. The estimated environmental NM concentrations are in the low range of ng to µg/L, except for Ti-based ones, which concentrations reach values on the order of mg/L. Empirical information regarding the ecotoxicity of environmental NM concentrations mainly focused on metal-based NM, however, it resulted poor and unbalanced in terms of materials and test species. Given its high predicted environmental concentration in comparison with the others, the ecotoxicity of Ti-based NM has been well assessed in algae and fish, while little is known regarding other NM types. While only a few marine species were addressed, the freshwater species Daphnia magna and Danio rerio accounted for the majority of studies on invertebrate and fish groups, respectively. Most of the reported responses are related to oxidative stress. Overall, we consider that invertebrate groups are the most vulnerable, with emphasis on microcrustaceans, as environmentally realistic metal-based NM concentration even caused mortality in some species. In the case of fish, we assumed that environmental concentrations of Ti-based NM represent a growing concern and threat; however, further studies should be carried out by employing other kinds of NM. Furthermore, more ecotoxicological information is needed in the case of carbon-based NM, as they are expected to considerably increase in terms of released amounts and applications in the near future.

Ecotoxicological effects, human and animal health risks of pollution and exposure to waste engine oils: a review.

Waste engine oils are hazardous waste oils originating from the transportation sector and industrial heavy-duty machinery operations. Improper handling, disposal, and miscellaneous misuses cause significant air, soil, sediments, surface water, and groundwater pollution. Occupational exposure by prolonged and repeated contact poses direct or indirect health risks, resulting in short-term (acute) or long-term (chronic) toxicities. Soil pollution causes geotoxicity by disrupting the biocenosis and physicochemical properties of the soil, and phytotoxicity by impairing plant growth, physiology and metabolism. Surface water pollution impacts aquatic ecosystems and biodiversity. Air pollution from incineration causes the release of greenhouse gases creating global warming, noxious gases and particulate matter eliciting pulmonary disorders. The toxicity of waste engine oil is due to the total petroleum hydrocarbons (TPH) composition, including polycyclic aromatic hydrocarbons (PAHs), benzene, toluene, ethylbenzene, xylene (BTEX), polychlorinated biphenyls (PCBs) congeners, organometallic compounds, and toxic chemical additives. The paper aims to provide a comprehensive overview of the ecotoxicological effects, human and animal health toxicology and exposure to waste engine oils. It highlights the properties and functions of engine oil and describes waste engine oil generation, disposal and recycling. It provides intensive evaluations and descriptions of the toxicokinetics, metabolism, routes of exposure and toxicosis in human and animal studies based on toxicological, epidemiological and experimental studies. It emphasises the preventive measures in occupational exposure and recommends risk-based remediation techniques to mitigate environmental pollution. The review will assist in understanding the potential risks of waste engine oil with significant consideration of the public health benefits and importance.

Eco-toxicological effects assessment: comparative characteristics of environmental conditions and status of vertebrate indicator species in the "Dnepr" launch vehicle accident zone.

Space rockets are regularly launched from the Baikonur Cosmodrome in Kazakhstan. The ecological impact of territories affected by the consequences of space rocket activities on the environment is a significant concern. On July 27, 2006, the Dnepr rocket was launched from the Baikonur Cosmodrome, resulting in an abnormal situation and its emergency fall. The rocket fuel components, heptyl, and amyl, led to decontamination and reclamation activities. However, the self-restoration of soil and vegetation cover is currently in the earliest stages. Consequently, an assessment of the remote consequences of the accidental fall of the Dnepr launch vehicle for environmental objects was conducted. The study involved eco-toxicological characterization of habitat conditions, assessment of species diversity, determination of key morphophenetic indicators, and the genetic status of indicator species as the main parameters. The results revealed an increased level of genotoxicity in soils at the accident site. While there were deviations from the norm in some morphophenetic and cytogenetic parameters in indicator animals, they were not critical for their existence in this territory. Possibly, at the physiological level, they exhibit a high enough adaptation potential and compensatory mechanism.

Ecotoxicological assessment, in freshwater environment, of wastewater sludge coupled and uncoupled with micro-polyvinyl chloride on algae and water fleas.

In the frame of bioeconomy and circular economy, wastewater sludge (WS) could be a good candidate for its use in agriculture as fertilizer, due to its high content of organic matter, N and P, but on the other hand, it is full of toxicants such as heavy metal, microplastics, detergent, antibiotics, and so on that can reach groundwater and water bodies in leachate form. In this study, we have investigated different sludge concentrations in the eluate form, combined and not with PVC on two different freshwater organisms Selenastrum capricornutum and Daphnia magna, using ecotoxicity tests. At the endpoint, we have evaluated inhibition growth rate, oxidative stress, and pigments production for S. capricornutum, while in case of D. magna, we have assessed organism immobilization and development. From our results, it emerged that at the higher WS concentration, there was not inhibition growth rate, while at oxidative stress, it was higher in algae treated with WS and PVC. Higher Chl-a production was shown for algae treated with 0.3 g/L of sludge coupled with PVC, where higher phaeopigments production were recorded for algae treated with 0.3 g/L of WS. D. magna has shown an opposite trend when compared with algae, where at the highest WS concentrations supplied was corresponding to an increased mortality explaned as the highest immobility percentage. PRACTITIONER POINTS: Wastewater sludge is used in agriculture as fertilizer. PVC microplastic presence and associate ecotoxicity was tested. PVC presence increased oxidative stress in S. capricornutum. D. magna was significantly affected by sludge concentrations supplied.

Ecotoxicological risk assessment on coagulation-flocculation in water/wastewater treatment: a systematic review.

It is undeniable that removal efficiency is the main factor in coagulation-flocculation (C-F) process for wastewater treatment. However, as far as environmental safety is concerned, the ecotoxicological aspect of the C-F process needs to be examined further. In this study, a systematic review was performed based on publications related to the toxicity research in C-F technology for wastewater treatment. Through a series of screening steps, available toxicity studies were categorized into four themes, namely acute toxicity, phytotoxicity, cytotoxicity, and genotoxicity, which comprised 48 articles. A compilation of the methodologies executed for each theme was also outlined. The findings show that conventional metallic coagulants (e.g., alum, iron chloride, and iron sulfate) were less toxic when tested on test species such as Daphnia magna (water flea), Lattuca sativa (lettuce), and animal cells compared to synthetic polymers. Natural coagulants such as chitosan or Moringa oleifera were less toxic compared to metallic coagulants; however, inconsistent results were observed. Moreover, an advanced C-F (electrocoagulation) as well as integration between C-F and Fenton, adsorption, and photocatalytic does not significantly change the toxicological profile of the system. It was found that diverse coagulants and flocculants, species sensitivity, complexity in toxicity testing, and dynamic environmental conditions were some key challenges faced in this field. Finally, it was expected that advances in technology, interdisciplinary collaboration, and a growing awareness of environmental sustainability will drive efforts to develop more effective and eco-friendly coagulants and flocculants, improve toxicity testing methodologies, and enhance the overall efficiency and safety of water and wastewater treatment processes.

Integrated assessment of the chemical, microbiological and ecotoxicological effects of a bio-packaging end-of-life in compost.

The present study aimed to i) assess the disintegration of a novel bio-packaging during aerobic composting (2 and 6 % tested concentrations) and evaluate the resulting compost ii) analyse the ecotoxicity of bioplastics residues on earthworms; iii) study the microbial communities during composting and in 'earthworms' gut after their exposure to bioplastic residues; iv) correlate gut microbiota with ecotoxicity analyses; v) evaluate the chemico-physical characterisation of bio-packaging after composting and earthworms' exposure. Both tested concentrations showed disintegration of bio-packaging close to 90 % from the first sampling time, and compost chemical analyses identified its maturity and stability at the end of the process. Ecotoxicological assessments were then conducted on Eisenia fetida regarding fertility, growth, genotoxic damage, and impacts on the gut microbiome. The bioplastic residues did not influence the earthworms' fertility, but DNA damages were measured at the highest bioplastic dose tested. Furthermore bioplastic residues did not significantly affect the bacterial community during composting, but compost treated with 2 % bio-packaging exhibited greater variability in the fungal communities, including Mortierella, Mucor, and Alternaria genera, which can use bioplastics as a carbon source. Moreover, bioplastic residues influenced gut bacterial communities, with Paenibacillus, Bacillus, Rhizobium, Legionella, and Saccharimonadales genera being particularly abundant at 2 % bioplastic concentration. Higher concentrations affected microbial composition by favouring different genera such as Pseudomonas, Ureibacillus, and Streptococcus. For fungal communities, Pestalotiopsis sp. was found predominantly in earthworms exposed to 2 % bioplastic residues and is potentially linked to its role as a microplastics degrader. After composting, Attenuated Total Reflection analysis on bioplastic residues displayed evidence of ageing with the formation of hydroxyl groups and amidic groups after earthworm exposure.

First evidence of lithium toxicity in the cryptic species complex of the estuarine copepod Eurytemora affinis.

The development of renewable and low-carbon energy sources means that strategic elements such as lithium (Li) are increasingly being used. The data available on the effects of Li on aquatic organisms are relatively scarce. The copepod Eurytemora affinis, widely distributed in the brackish estuarine waters of the northern hemisphere, is a species of choice in ecotoxicology but in fact constitutes a cryptic species complex, composed of at least six cryptic species. Cryptic diversity can lead to misinterpretation and alter the reproducibility of routine ecotoxicological tests. In the present study, two cryptic species of the E. affinis complex from the Seine (European clade) and the St. Lawrence (North-Atlantic clade) estuaries were used to assess Li toxicity and to compare their differential sensitivity. Larvae were exposed to different concentrations of Li (0.4, 4.39, 35.36 and 80.83 mg L-1) under semi-static conditions for 96 h. Larval development stages were determined and log-logistic functions were fitted to evaluate mortality (LC50) and growth (EC50) parameters. After 96 h of exposure, the results showed that the European and North-Atlantic clades had LC50 values of 55.33 and 67.81 mg L-1 and EC50 values of 28.94 and 41.45 mg L-1, respectively. A moderate difference in sensitivity to Li between the European and North-Atlantic clades of the E. affinis complex was observed. Thus, the cryptic species diversity should be considered using E. affinis to avoid bias in the interpretation of the data. Despite environmental concentrations of Li are expected to increase over the next years, EC50 and LC50 found for E. affinis cryptic species are largely higher than Li environmental concentrations to provoke extreme effects.

From a validated targeted method to a retrospective UHPLC-HRMS non-targeted analysis unveiling COVID-19-related contaminants in clams. Have we bias in marine model organisms for ecotoxicological studies?

This study explores a retrospective non-targeted analysis (NTA), based on Ultra High-Performance Liquid Chromatography coupled to High-Resolution Mass Spectrometry (UHPLC-HRMS), to assess hidden chemicals of emerging concern (CECs) in marine model organisms. Conventional ecotoxicological studies do not include evaluating the natural habitats of the collected organisms, missing the possibility of highlighting unexpected pollutants, and thus compromising the correctness and reliability of the experimental results. In this paper we reprocessed samples previously collected from the Venice Lagoon for ecotoxicological studies and used for targeted analysis of three bisphenols-related compounds (i.e. BPS, BPF and BPAF) on seawater and specimens of the clam Ruditapes philippinarum. Results from the validation were the following: accuracy, expressed as percentage recoveries (R%), in the range 80%

Ecotoxicological, ecological, and human health risks of total carbohydrates and some inorganic pollutants on the Nile Delta region along the Egyptian Mediterranean Coast.

This is one of few studies dealing with the potential impact of total carbohydrates (TCHO), and some inorganic pollutants (F, B, As, V, Se) on human health. Additionally, the latter pollutants toxicological and ecological effects on the Egyptian Mediterranean Coast, especially, the Nile Delta region, were investigated. Both F (0.18 ± 0.09 mg/g) and As (2.47 ± 5.39 µg/g) were of lower concentrations compared to previous reports. Values of all ecological and ecotoxicity indices, particularly, the risk quotient (RQ), showed that arsenic had the most adverse biological effects on three trophic levels (algae, invertebrates, and fish). Children and adults non-carcinogenic hazard index (HI) values were <1, revealing that sediments in the studied area would pose no risk to humans. However, arsenic carcinogenic risk (CR) values exceeded the maximum permissible limits, implying risk to children and adults. These findings could anticipate toxic impacts of polluted effluents on the Nile Delta region.

Multi-omics integration analysis: Tools and applications in environmental toxicology.

Nowadays, traditional single-omics study is not enough to explain the causality between molecular alterations and toxicity endpoints for environmental pollutants. With the development of high-throughput sequencing technology and high-resolution mass spectrometry technology, the integrative analysis of multi-omics has become an efficient strategy to understand holistic biological mechanisms and to uncover the regulation network in specific biological processes. This review summarized sample preparation methods, integration analysis tools and the application of multi-omics integration analyses in environmental toxicology field. Currently, omics methods have been widely applied being as the sensitivity of early biological response, especially for low-dose and long-term exposure to environmental pollutants. Integrative omics can reveal the overall changes of genes, proteins, and/or metabolites in the cells, tissues or organisms, which provide new insights into revealing the overall toxicity effects, screening the toxic targets, and exploring the underlying molecular mechanism of pollutants.

Derivation of ecotoxicologically acceptable Cu concentrations in soil from different land uses in South Korea.

This study aimed to establish ecotoxicologically acceptable Cu concentrations for soil-residing species by integrating the biotic ligand model and the species sensitivity distribution. Statistical analyses were performed on 35 soil solution samples collected from four distinct land use sites: residential, agricultural, forested, and industrial regions. The environmental parameters of these samples, including pH, dissolved organic carbon (DOC), Ca2⁺, Mg2⁺, K⁺, and Na⁺ concentrations, exhibited wide variations across the four regions. Specifically, pH and the concentrations of Mg2⁺, K⁺, and Na⁺ showed significant variability. Additionally, a strong correlation was observed between pH and Ca2⁺, as well as between the DOC concentration and Mg2⁺ and Na⁺. Using the biotic ligand model, we derived the half-maximal effective activities of Cu (EC50{Cu2+}) for 10 soil organisms based on the chemical compositions of the soil solution samples. Additionally, a species sensitivity distribution approach was employed to determine the 5% hazardous concentration (HC5) for soil biota, which was closely associated with DOC and Na⁺ concentrations, with Mg2⁺ playing a secondary role. We attributed these relationships to the formation of DOC complexes that mitigate Cu toxicity, along with competitive interactions with cations. Notably, HC5 values did not differ significantly across sampling sites (p = 0.523). Clustering based on environmental factors grouped the samples into four clusters, each containing soils from different land use types. However, the third cluster included an outlier from agricultural soil due to its unusually high pH and DOC levels. These findings suggest that it is crucial to consider site-specific soil characteristics when determining ecotoxicologically acceptable Cu concentrations, and soil solution characteristics do not always align with specific land use patterns.

ECOTOXr: An R package for reproducible and transparent retrieval of data from EPA's ECOTOX database.

The US EPA ECOTOX database provides key ecotoxicological data that are crucial in environmental risk assessment. It can be used for computational predictions of toxicity or indications of hazard in a wide range of situations. There is no standardised or formalised method for extracting and subsetting data from the database for these purposes. Consequently, results in such meta-analyses are difficult to reproduce. The present study introduces the software package ECOTOXr, which provides the means to formalise data retrieval from the ECOTOX database in the R scripting language. Three cases are presented to evaluate the performance of the package in relation to earlier data extractions and searches on the website. These cases demonstrate that the package can reproduce data sets relatively well. Furthermore, they illustrate how future studies can further improve traceability and reproducibility by applying the package and adhering to some simple guidelines. This contributes to the FAIR principles, credibility and acceptance of research that uses data from the ECOTOX database.

Microbiological toxicity tests using standardized ISO/OECD methods-current state and outlook.

Microbial toxicity tests play an important role in various scientific and technical fields including the risk assessment of chemical compounds in the environment. There is a large battery of normalized tests available that have been standardized by ISO (International Organization for Standardization) and OECD (Organization for Economic Co-operation and Development) and which are worldwide accepted and applied. The focus of this review is to provide information on microbial toxicity tests, which are used to elucidate effects in other laboratory tests such as biodegradation tests, and for the prediction of effects in natural and technical aqueous compartments in the environment. The various standardized tests as well as not normalized methods are described and their advantages and disadvantages are discussed. In addition, the sensitivity and usefulness of such tests including a short comparison with other ecotoxicological tests is presented. Moreover, the far-reaching influence of microbial toxicity tests on biodegradation tests is also demonstrated. A new concept of the physiological potential of an inoculum (PPI) consisting of microbial toxicity tests whose results are expressed as a chemical resistance potential (CRP) and the biodegradation adaptation potential (BAP) of an inoculum is described that may be helpful to characterize inocula used for biodegradation tests. KEY POINTS: • Microbial toxicity tests standardized by ISO and OECD have large differences in sensitivity and applicability. • Standardized microbial toxicity tests in combination with biodegradability tests open a new way to characterize inocula for biodegradation tests. • Standardized microbial toxicity tests together with ecotoxicity tests can form a very effective toolbox for the characterization of toxic effects of chemicals.

Ecotoxicological evaluation of an aqueous phytoextract of Melia azedarach L.

Melia azedarach L. is a Meliaceae that has shown important insecticidal activities. However, few researchers have extensively studied the toxicology of aqueous extracts of M. azedarach (MAE). Therefore, the main objective of this study was to characterize the phyto-eco-toxicological profile of MAE. First, a botanical and phytochemical characterization of MAE was performed using a histological, and metabolomic multi-analytical approach. Second, the toxicological effects on pollinating insects (Apis mellifera ligustica) and soil collembola (Folsomia candida) were evaluated. In addition, acute toxicity was evaluated in zebrafish (Danio rerio) to assess effects on aquatic fauna, and toxicity was determined in human neuroblastoma (SH-SY5Y) and fibroblast (FB-21) cell models. Finally, phytotoxic effects on germination of Cucumis sativus L., Brassica rapa L. and Sorghum vulgare L. were considered. Metabolomic analyses revealed the presence of not only limonoids but also numerous alkaloids, flavonoids and terpenoids in MAE. Histological analyses allowed us to better localize the areas of leaf deposition of the identified secondary metabolites. Regarding the ecotoxicological data, no significant toxicity was observed in bees and collembola at all doses tested. In contrast, severe cardiac abnormalities were observed in zebrafish embryos at concentrations as low as 25 µg/mL. In addition, MAE showed toxicity at 1.6 µg/mL and 6.25 µg/mL in FB-21 and SH-SY5Y cells, respectively. Finally, MAE inhibited seed germination with inhibitory concentrations starting from 5.50 µg/mL in B. rapa, 20 µg/mL in S. vulgare, and 31 µg/mL in C. sativus. Although M. azedarach extracts are considered valuable natural insecticides, their ecological impact cannot be underestimated. Even the use of an environmentally friendly solvent (an aqueous solution), for the first time, is not without side effects. Therefore, the data collected in this study show the importance of evaluating the dosages, modes of administration and production methods of M. azedarach phytoextracts in agricultural settings.

Silence is not always golden: A closer look at potential environmental and ecotoxicological impacts of large-scale dsRNA application.

RNA interference (RNAi) technology has emerged as a pivotal strategy in sustainable pest management, offering a targeted approach that significantly mitigates the environmental and health risks associated with traditional insecticides. Originally implemented through genetically modified organisms (GMOs) to produce specific RNAi constructs, the technology has evolved in response to public and regulatory concerns over GMOs. This evolution has spurred the development of non-transgenic RNAi applications such as spray-induced gene silencing (SIGS), which employs double-stranded RNA (dsRNA) to silence pest genes directly without altering the plant's genetic makeup. Despite its advantages in specificity and reduced ecological footprint, SIGS faces significant obstacles, particularly the instability of dsRNA in field conditions, which limits its practical efficacy. To overcome these limitations, innovative delivery mechanisms have been developed. These include nanotechnology-based systems, minicells, and nanovesicles, which are designed to protect dsRNA from degradation and enhance its delivery to target organisms. While these advancements have improved the stability and application efficiency of dsRNA, comprehensive assessments of their environmental safety and the potential for increased exposure risks to non-target organisms remain incomplete. This comprehensive review aims to elucidate the environmental fate of dsRNA and evaluate the potential risks associated with its widespread application on non-target organisms, encompassing soil microorganisms, beneficial insects, host plants, and mammals. The objective is to establish a more refined framework for RNAi risk assessment within environmental and ecotoxicological contexts, thereby fostering the development of safer, non-transgenic RNAi-based pest control strategies.

Transferring knowledge across aquatic species via clustering techniques to unravel patterns of pesticide toxicity.

In silico modelling takes the advantage of accelerating ecotoxicological assessments on hazardous chemicals without conducting risky in vivo experiments under ethic regulation. To date, the prevailing strategy of one model for one species cannot be well generalized to multi-species modelling. In this work, we propose a new strategy of one model for multiple species to facilitate knowledge transfer across aquatic species. The available lethal concentration values of 4952 pesticides on 651 fish species are aggregated into one toxicity response matrix, purely through which we attempt to unravel fish toxicosis-phylogenesis relationships and pesticide toxicity-structure relationships via clustering techniques including non-negative matrix factorization (NMF) and hierarchical clustering. The clustering results suggest that (1) close NMF weights indicate close species-toxicosis and pesticide-toxicity profiles; (2) and that species toxicosis patterns are related with species phylogenetic relationships; (3) and that close pesticide-toxicity profiles indicate similar atom-pair structural fingerprints. These environmental, chemical and biological insights can be used as expert knowledge for environmentalists to manually gain knowledge about untested species/pesticides from tested species/pesticides, and meanwhile provide support for us to build in silico models from species phylogenetic and pesticide structural points of view. Besides unravelling the mechanisms behind toxicity response, we also adopt stratified cross validation and external test to validate the reliability of using NMF to predict missing toxicity values. Independent test on external data shows that NMF achieves 0.8404-0.9397 R2 on four fish species. In the context of toxicity prediction, non-negative matrix factorization can be viewed as a model based on quantitative activity-activity relationships (QAAR), and provides an alternative approach of inferring toxicity values on untested species from tested species.