Bio-QSARs 2.0: Unlocking a new level of predictive power for machine learning-based ecotoxicity predictions by exploiting chemical and biological information.

Practical, legal, and ethical reasons necessitate the development of methods to replace animal experiments. Computational techniques to acquire information that traditionally relied on animal testing are considered a crucial pillar among these so-called new approach methodologies. In this light, we recently introduced the Bio-QSAR concept for multispecies aquatic toxicity regression tasks. These machine learning models, trained on both chemical and biological information, are capable of both cross-chemical and cross-species predictions. Here, we significantly extend these models' applicability. This was realized by increasing the quantity of training data by a factor of approximately 20, accomplished by considering both additional chemicals and aquatic organisms. Additionally, variable test durations and associated random effects were accommodated by employing a machine learning algorithm that combines tree-boosting with mixed-effects modeling (i.e., Gaussian Process Boosting). We also explored various biological descriptors including Dynamic Energy Budget model parameters, taxonomic distances, as well as genus-specific traits and investigated the inclusion of mode-of-action information. Through these efforts, we developed Bio-QSARs for fish and aquatic invertebrates with exceptional predictive power (R squared of up to 0.92 on independent test sets). Moreover, we made considerable strides to make models applicable for a range of use cases in environmental risk assessment as well as research and development of chemicals. Models were made fully explainable by implementing an algorithmic multicollinearity correction combined with SHapley Additive exPlanations. Furthermore, we devised novel approaches for applicability domain construction that take feature importance into account. We are hence confident these models, which are available via open access, will make a significant contribution towards the implementation of new approach methodologies and ultimately have the potential to support "Green Chemistry" and "Green Toxicology".

Temperature induced changes in species distribution increases sensitivity of aquatic invertebrate communities to chemicals.

In this commentary, I will discuss how climate warming might influence the impacts of chemicals on (aquatic) ecosystems. It provides a commentary on Sinclair et al. (2024).

Microplastics alter the leaf litter breakdown rates and the decomposer community in subtropical lentic microhabitats.

Microplastics, pervasive pollutants in aquatic environments, have been primarily studied for their impact on marine ecosystems. However, their effects on freshwater systems, particularly in forested phytotelmata habitats, remain understudied in Subtropical systems. This research examines the influence of varying microplastic concentrations (0.0, 200, 2,000, 20,000, and 200,000 ppm) on leaf litter breakdown of Inga vera (in bags of 10 and 0.05 mm mesh) and the naturally associated invertebrate community occurring in forested phytotelmata. The study employs an experimental design with microplastic concentration treatments in artificial microcosms (buckets with 800 mL of rainwater) arranged in an area of Atlantic Rain Forest native vegetation of Subtropical systems. The results indicate that elevated concentrations of microplastics may enhance leaf litter breakdown (6-8%), irrespective of the bag mesh, attributed to heightened decomposer activity and biofilm formation. Consequently, this contributes to increased invertebrate richness (33-37%) and greater shredder abundance (21-37%). Indicator analysis revealed that Culicidae, Stratiomyidae, Chironomidae, Empididae, Planorbidae, and Ceratopogonidae were indicative of some microplastic concentrations. These findings underscore the significance of accounting for microplastics when evaluating the taxonomic and trophic characteristics of invertebrate communities, as well as the leaf breakdown process in Subtropical systems.

Organophosphate pesticide residue impact on water quality and changes in macroinvertebrate community in an Afrotropical stream flowing through farmlands.

River Chanchaga has experienced significant agricultural practices around its catchment, which involved the indiscriminate use of pesticides. However, residents of the study area are not well aware of the negative impact of pesticides on water quality and macroinvertebrates. In this study, the first report on the influence of organophosphate pesticide contamination on the abundance of the macroinvertebrate community was provided. Sampling for the determination of organophosphate pesticide residues was carried out during the peak of the two seasons, while macroinvertebrates and physicochemical variables were observed for 6 months. We examined 11 organophosphate pesticide residues using gas chromatography coupled with mass spectrometry, 12 water quality variables, and 625 macroinvertebrate individuals. The concentration of recorded organophosphate pesticide residues ranged from 0.01 to 0.52 µg/L. From the Canonical Correspondence Analysis plot, malathion, chlorine, and paraffin show a positive correlation with Unima sp., Hydrocanthus sp., Chironomus sp., and Potadoma sp. At station 3, depth shows a positive correlation with Biomphalaria sp. and Zyxomma sp., indicating poor water quality as most of these macroinvertebrates are indicators of water pollution. Diuron and carbofuran show a negative correlation with Lestes sp. and Pseudocloeon sp., and these are pollution-sensitive macroinvertebrates. The total mean concentration of organophosphate pesticide residues was above international drinking water standards set by the World Health Organization except for paraffin, chlorpyrifos, and diuron. In conclusion, the observations recorded from this research are useful in managing pesticide applications around the river catchment.

Developmental toxicity of pre-production plastic pellets affects a large swathe of invertebrate taxa.

Microplastics pose risks to marine organisms through ingestion, entanglement, and as carriers of toxic additives and environmental pollutants. Plastic pre-production pellet leachates have been shown to affect the development of sea urchins and, to some extent, mussels. The extent of those developmental effects on other animal phyla remains unknown. Here, we test the toxicity of environmental mixed nurdle samples and new PVC pellets for the embryonic development or asexual reproduction by regeneration of animals from all the major animal superphyla (Lophotrochozoa, Ecdysozoa, Deuterostomia and Cnidaria). Our results show diverse, concentration-dependent impacts in all the species sampled for new pellets, and for molluscs and deuterostomes for environmental samples. Embryo axial formation, cell specification and, specially, morphogenesis seem to be the main processes affected by plastic leachate exposure. Our study serves as a proof of principle for the potentially catastrophic effects that increasing plastic concentrations in the oceans and other ecosystems can have across animal populations from all major animal superphyla.

Safe and sustainable by design Ag nanomaterials: A case study to evaluate the bio-reactivity in the environment using a soil model invertebrate.

Safe-and-sustainable-by-design (SSbD) nanomaterials (NMs) or NM-containing products are a priority. Silver (Ag) NMs have a vast array of applications, including biomedical and other products, even as nanopesticides. Thus, their release to the environment is expected to increase. The aim of the present study was to assess the ecotoxicity of the SSbD Ag NM to the soil model species Enchytraeus crypticus (Oligochaeta). The Ag NM tested consists in a SSbD Ag with biomedical applications, a hydroxyethyl cellulose (HEC) coated Ag NMs (AgHEC) and its toxicity was compared to the naked Ag NMs (Ag-Sigma), an Ag-based biomedical product (PLLA-Ag: Poly l-Lactide microfibers doped with Ag), and AgNO3. Effects were assessed both in soil and aqueous media, following the standard OECD guideline in soil (28 days) and the OECD extension (56 days), and short-term pulse (5 days) in aqueous media: reconstituted water (ISO water) and soil:water (S:W) extracts, followed by a 21-days recovery period in soil. Ag materials were thoroughly characterized as synthesized and during the test in media and animals. Results in S:W showed AgHEC was more toxic than Ag-Sigma (ca. 150 times) and PLLA-Ag (ca. 2.5 times), associated with a higher Ag uptake. Higher toxicity was related to a smaller hydrodynamic size and higher suspension stability, which in turn resulted in a higher bioavailability of Ag NMs and released ions, particularly in S:W. Toxicity was correlated with the main physicochemical features, providing useful prediction of AgNMs bioactivity. The ability to test E. crypticus in a range of media with different and/or increasing complexity (water, S:W extracts, soil) provided an excellent source to interpret results and is here recommended.

Toxicity of 6PPD-quinone to four freshwater invertebrate species.

The antioxidant N-(1,3-Dimethylbutyl)-N'-phenyl-p- phenylenediamine (6PPD) is used to protect the rubber in tires from oxidation, which extends the life of the tire. When oxidized, 6PPD is transformed into 6PPD-quinone (6PPDQ). 6PPDQ, along with other tire ingredients, can enter aquatic ecosystems through the transport of tire wear particles in runoff during a precipitation event. The mass mortality of coho salmon following precipitation events in urban areas lead to the discovery that 6PPDQ is the likely cause due to coho salmon's relatively high sensitivity to 6PPDQ. The assessment of 6PPDQ toxicity to other aquatic species has expanded, but it has focused on fish. This study investigated the toxicity of 6PPDQ to four freshwater invertebrate species, larval burrowing mayfly (Hexagenia spp.), juvenile cladoceran (Daphnia magna), file ramshorn snail embryo (Planorbella pilsbryi), and adult washboard mussel (Megalonaias nervosa). For all four species, the highest concentration of 6PPDQ tested did not result in significant mortality. This translated into the determination of the highest concentration that did not cause significant mortality (NOEC) for Hexagenia spp., D. magna, P. pilsbryi, and M. nervosa of 232.0, 42.0, 11.7, and 17.9 µg/L, respectively. The data from this study indicate that freshwater invertebrates are not as sensitive to 6PPDQ as some salmonid species (e.g., coho salmon Oncorhynchus kisutch). This study also analyzed 6PPDQ in road runoff from around the city of Guelph in Ontario, Canada. 6PPQ was detected in all samples but the concentration was two orders of magnitude lower than the NOECs for the four tested species of freshwater invertebrate.

Impacts of urban and industrial pollution on functional traits of benthic macroinvertebrates: Are some traits advantageous for survival?

Urbanization and industrialization produce substantial changes in biodiversity and in the functionality of ecosystems. However, little is known about how anthropic pressures might drive these changes and about their functional consequences. We aimed to determine the responses of macroinvertebrate biological traits to urban and industrial pollution and assess the impacts of these disturbances on the functional diversity of these assemblages. We sampled benthic macroinvertebrates in 27 sites of four basins with different urban disturbance gradients (rural, peri-urban, and urban-industrial), among them the Matanza-Riachuelo River, one of the most polluted basins in the world. We classified macroinvertebrates into 11 traits and 56 categories. Then, we performed an RLQ analysis and computed functional richness, evenness, divergence and Rao diversity indexes for each site and community weighted means for each trait category. The urban and industrial sites (mainly low and middle Matanza-Riachuelo River Basin) showed high concentrations of ammonium, SRP, conductivity, COD, BOD, and organic matter, as well as the lowest values of DO. The functional richness and Rao index of these sites were significantly lower than that of the other sites. Macroinvertebrate traits associated with urban and industrial sites were aerial respiration (spiracles), forms of resistance (eggs or statoblast), cylindrical body shape, oviparity, feeding on microinvertebrates, and full water swimmers. These traits potentially enabled tolerant species persistence at polluted sites while gills, grazers, and crawlers were sensitive to these disturbances. Urban and industrial activities influence biological traits, producing the disappearance or dominance of certain traits in macroinvertebrate assemblages. As a consequence, extreme pollution caused predictable trait-based community changes resulting in reduced functional diversity, and potentially altered the ecosystem function.

Ethylhexyl methoxycinnamate and butyl methoxydibenzoylmethane: Toxicological effects on marine biota and human concerns.

Ethylhexyl methoxycinnamate (EHMC) (CAS number: 5466-77-3) and butyl methoxydibenzoylmethane (BMDM) (CAS number: 70356-09-1) are important sunscreens. However, frequent application of large amounts of these compounds may reflect serious environmental impact, once it enters the environment through indirect release via wastewater treatment or immediate release during water activities. In this article, we reviewed the toxicological effects of EHMC and BMDM on aquatic ecosystems and the human consequences. According to the literature, EHMC and BMDM have been detected in water samples and sediments worldwide. Consequently, these compounds are also present in several marine organisms like fish, invertebrates, coral reefs, marine mammals, and other species, due to its bioaccumulation potential. Studies show that these chemicals are capable of damaging the aquatic beings in different ways. Further, bioaccumulation studies have shown that EHMC biomagnifies through trophic levels, which makes human seafood consumption a concern because the higher position in the trophic chain, the more elevate levels of ultraviolet (UV) filters are detected, and it is established that EHMC present adverse effects on the human organism. In contrast, there are no studies on the BMDM bioaccumulation and biomagnification potential. Different strategies can be adopted to avoid the damage caused by sunscreens in the environment and human organism. Two of them include the use of natural photoprotectors, such as polyphenols, in association with UV filters in sunscreens and the development of new and safer UV filters. Overall, this review shows the importance of studying the impacts of sunscreens in nature and developing safer sunscreens and formulations to safeguard marine fauna, ecosystems, and humans.

Alternative animal models in predictive toxicology.

Toxicity testing relies heavily on animals, especially rodents as part of the non-clinical laboratory testing of substances. However, the use of mammalians and the number of animals employed in research has become a concern for institutional ethics committees. Toxicity testing involving rodents and other mammals is laborious and costly. Alternatively, non-rodent models are used as replacement, as they have less ethical considerations and are cost-effective. Of the many alternative models that can be used as replacement models, which ones can be used in predictive toxicology? What is the correlation between these models and rodents? Are there standardized protocols governing the toxicity testing of these commonly used predictive models? This review outlines the common alternative animal models for predictive toxicology to address the importance of these models, the challenges, and their standard testing protocols.

Effects of Pesticides on Longevity and Bioenergetics in Invertebrates-The Impact of Polyphenolic Metabolites.

Environmentally hazardous substances such as pesticides are gaining increasing interest in agricultural and nutritional research. This study aims to investigate the impact of these compounds on the healthspan and mitochondrial functions in an invertebrate in vivo model and in vitro in SH-SY5Y neuroblastoma cells, and to investigate the potential of polyphenolic metabolites to compensate for potential impacts. Wild-type nematodes (Caenorhabditis elegans, N2) were treated with pesticides such as pyraclostrobin (Pyr), glyphosate (Gly), or fluopyram (Fluo). The lifespans of the nematodes under heat stress conditions (37 °C) were determined, and the chemotaxis was assayed. Energetic metabolites, including adenosine triphosphate (ATP), lactate, and pyruvate, were analyzed in lysates of nematodes and cells. Genetic expression patterns of several genes associated with lifespan determination and mitochondrial parameters were assessed via qRT-PCR. After incubation with environmentally hazardous substances, nematodes were incubated with a pre-fermented polyphenol mixture (Rechtsregulat®Bio, RR) or protocatechuic acid (PCA) to determine heat stress resistance. Treatment with Pyr, Glyph and Fluo leads to dose-dependently decreased heat stress resistance, which was significantly improved by RR and PCA. The chemotaxes of the nematodes were not affected by pesticides. ATP levels were not significantly altered by the pesticides, except for Pyr, which increased ATP levels after 48 h leads. The gene expression of healthspan and mitochondria-associated genes were diversely affected by the pesticides, while Pyr led to an overall decrease of mRNA levels. Over time, the treatment of nematodes leads to a recovery of the nematodes on the mitochondrial level but not on stress resistance on gene expression. Fermented extracts of fruits and vegetables and phenolic metabolites such as PCA seem to have the potential to recover the vitality of C. elegans after damage caused by pesticides.

Physiological Effects of Neonicotinoid Insecticides on Non-Target Aquatic Animals-An Updated Review.

In this paper, we review the effects of large-scale neonicotinoid contaminations in the aquatic environment on non-target aquatic invertebrate and vertebrate species. These aquatic species are the fauna widely exposed to environmental changes and chemical accumulation in bodies of water. Neonicotinoids are insecticides that target the nicotinic type acetylcholine receptors (nAChRs) in the central nervous systems (CNS) and are considered selective neurotoxins for insects. However, studies on their physiologic impacts and interactions with non-target species are limited. In researches dedicated to exploring physiologic and toxic outcomes of neonicotinoids, studies relating to the effects on vertebrate species represent a minority case compared to invertebrate species. For aquatic species, the known effects of neonicotinoids are described in the level of organismal, behavioral, genetic and physiologic toxicities. Toxicological studies were reported based on the environment of bodies of water, temperature, salinity and several other factors. There exists a knowledge gap on the relationship between toxicity outcomes to regulatory risk valuation. It has been a general observation among studies that neonicotinoid insecticides demonstrate significant toxicity to an extensive variety of invertebrates. Comprehensive analysis of data points to a generalization that field-realistic and laboratory exposures could result in different or non-comparable results in some cases. Aquatic invertebrates perform important roles in balancing a healthy ecosystem, thus rapid screening strategies are necessary to verify physiologic and toxicological impacts. So far, much of the studies describing field tests on non-target species are inadequate and in many cases, obsolete. Considering the current literature, this review addresses important information gaps relating to the impacts of neonicotinoids on the environment and spring forward policies, avoiding adverse biological and ecological effects on a range of non-target aquatic species which might further impair the whole of the aquatic ecological web.

Lethal and sub-lethal activity of Brevibacillus laterosporus on the mosquito Aedes albopictus and side effects on non-target water-dwelling invertebrates.

The biocidal potential of Brevibacillus laterosporus against mosquitoes of major medical importance has been widely documented, but its effects on non-target invertebrates are still poorly known. In this study, we determined the lethal and sub-lethal effects of B. laterosporus strain UNISS 18, an entomopathogenic bacterium known for its effectiveness against synanthropic Diptera, on the larvae of the Asian tiger mosquito Aedes albopictus, a vector of several pathogens to humans. Moreover, we compared the larvicidal activity with the lethal action on the invasive snail Physella acuta and on two non-target water-dwelling species: the mayfly Cloeon dipterum, and the harlequin fly Chironomus riparius. B. laterosporus exhibited significant lethal effects on all the tested species with a concentration-dependent activity. However, the susceptibility varied among species, with a higher susceptibility of Ae. albopictus (LC50 = 0.16 × 107 spores mL-1) than the other species (LC50 = 0.31, 0.33, and 0.30 × 107 spores mL-1 for C. dipterum, C. riparius, and P. acuta, respectively). While 1st instar mosquito larvae were very susceptible to the bacterial infection, no effects on preimaginal development stages and adult emergence were observed at sub-lethal spores' concentrations. Even if the efficacy of B. laterosporus against Ae. albopictus and the invasive freshwater snail P. acuta is promising for their control, the susceptibility of non-target beneficial aquatic insects, highlights the need of accurate evaluations before applying B. laterosporus for pest management in water environments.

Oxytocin Involvement in Body Composition Unveils the True Identity of Oxytocin.

The origin of the Oxytocin/Vasopressin system dates back about 600 million years. Oxytocin (Oxt) together with Vasopressin (VP) regulate a diversity of physiological functions that are important for osmoregulation, reproduction, metabolism, and social behavior. Oxt/VP-like peptides have been identified in several invertebrate species and they are functionally related across the entire animal kingdom. Functional conservation enables future exploitation of invertebrate models to study Oxt's functions not related to pregnancy and the basic mechanisms of central Oxt/VP signaling. Specifically, Oxt is well known for its effects on uteri contractility and milk ejection as well as on metabolism and energy homeostasis. Moreover, the striking evidence that Oxt is linked to energy regulation is that Oxt- and Oxytocin receptor (Oxtr)-deficient mice show late onset obesity. Interestingly Oxt-/- or Oxtr-/- mice develop weight gain without increasing food intake, suggesting that a lack of Oxt reduce metabolic rate. Oxt is expressed in a diversity of skeletal muscle phenotypes and regulates thermogenesis and bone mass. Oxt may increases skeletal muscle tonicity and/or increases body temperature. In this review, the author compared the three most recent theories on the effects of Oxt on body composition.

Response of aquatic macroinvertebrates communities to multiple anthropogenic stressors in a lowland tributary river.

In the current study the response of aquatic macroinvertebrate communities to multiple anthropogenic stressors in a typical lowland river that crosses pristine sectors situated toward headwaters, as well as densely populated urban areas was assessed. We wished to develop an effective bioassay procedure for assessing water and sediment quality in lotic ecosystems from Romania with the aid of macroinvertebrate organisms correlated with physico-chemical parameters and pollutants in both dissolved fractions and material bonded to sediment. A fast scanning approach of the river, from springs to the mouth, was employed. We found significant changes in physico-chemical parameters along a longitudinal gradient, the highest values being registered within the urban area and heavily agriculturally developed areas. The macroinvertebrates showed affinities for certain abiotic factors, emphasising their potential use for future studies as reliable ecological indicators, shaped by a synergic combination of urban effects and magnitude of type of land use.

The neonicotinoid alternative sulfoxaflor causes chronic toxicity and impairs mitochondrial energy production in Chironomus kiinensis.

Unintentional environmental consequences caused by neonicotinoids reinforce the development of safer alternatives. Sulfoxaflor is considered such an alternative. However, ecological risk of sulfoxaflor remains largely unknown. Here, we investigated the acute and chronic toxicity of sulfoxaflor to a benthic invertebrate, Chironomus kiinensis. Sulfoxaflor showed lower lethality than imidacloprid to midges, with LC50 values of 84.1 (81.5-87.3), 66.3 (34.8-259), and 47.5 (29.5-306) µg/L for 96-h, 10-d, and 23-d exposures, respectively. Conversely, sulfoxaflor significantly inhibited C. kiinensis growth and emergence in chronic exposures when concentrations were above 20 µg/L. Effects on energy production were assessed through in vitro tests using mitochondria isolated from C. kiinensis. Sulfoxaflor disrupted mitochondrial state-3 respiration, meanwhile, adenosine triphosphatase (ATPase) activity and adenosine triphosphate (ATP) production were both inhibited in a dose-dependent manner. The observed mitochondrial dysfunction may be related to the decreased organismal growth and emergence, which could further influence biodiversity. Interestingly, sulfoxaflor uptake in C. kiinensis was detected even after emergence, implying its potential to be transported along food webs and among environmental compartments. This study provides thorough investigations on the toxicity of an emerging neonicotinoid alternative to Chironomidae. Data derived from the current study are useful to inform future ecological risk assessment and benefit problem-solving to the overall agriculture-environment nexus.

Applied pesticide toxicity shifts toward plants and invertebrates, even in GM crops.

Pesticide impacts are usually discussed in the context of applied amounts while disregarding the large but environmentally relevant variations in substance-specific toxicity. Here, we systemically interpret changes in the use of 381 pesticides over 25 years by considering 1591 substance-specific acute toxicity threshold values for eight nontarget species groups. We find that the toxicity of applied insecticides to aquatic invertebrates and pollinators has increased considerably-in sharp contrast to the applied amount-and that this increase has been driven by highly toxic pyrethroids and neonicotinoids, respectively. We also report increasing applied toxicity to aquatic invertebrates and pollinators in genetically modified (GM) corn and to terrestrial plants in herbicide-tolerant soybeans since approximately 2010. Our results challenge the claims of a decrease in the environmental impacts of pesticide use.

Effects of a bacteria-produced algicide on non-target marine invertebrate species.

Harmful algal blooms (HABs) affect both freshwater and marine systems. Laboratory experiments suggest an exudate produced by the bacterium Shewanella sp. IRI-160 could be used to prevent or mitigate dinoflagellate blooms; however, effects on non-target organisms are unknown. The algicide (IRI-160AA) was tested on various ontogenetic stages of the copepod Acartia tonsa (nauplii and adult copepodites), the blue crab Callinectes sapidus (zoea larvae and megalopa postlarvae), and the eastern oyster Crassostrea virginica (pediveliger larvae and adults). Mortality experiments with A. tonsa revealed that the 24-h LC50 was 13.4% v/v algicide for adult females and 5.96% for early-stage nauplii. For C. sapidus, the 24-h LC50 for first-stage zoeae was 16.8%; results were not significant for megalopae or oysters. Respiration rates for copepod nauplii increased in the 11% concentration, and in the 11% and 17% concentrations for crab zoeae; rates of later stages and oysters were unaffected. Activity level was affected for crab zoeae in the 1%, 11%, and 17% treatments, and for oyster pediveliger larvae at the 17% level. Activity of later stages and of adult copepods was unaffected. Smaller, non-target biota with higher surface to volume could be negatively impacted from IRI-160AA dosing, but overall the taxa and stages assayed were tolerant to the algicide at concentrations required for dinoflagellate mortality (EC50 = ~ 1%).

Toxic Effects of Fine Plant Powder Impregnated With Avermectins on Mosquito Larvae and Nontarget Aquatic Invertebrates.

The toxic effects of an avermectin-impregnated fine plant powder (AIFP) against larval Aedes aegypti L. (Diptera: Culicidae), Culex modestus Ficalbi (Diptera: Culicidae), and Anopheles messeae Falleroni (Diptera: Culicidae), as well as selected nontarget aquatic invertebrates, were studied under laboratory conditions. The possibility of trophic transfer of avermectins (AVMs) through the food chain and their toxic effects on predaceous species fed AIFP-treated mosquito larvae was also evaluated. Among mosquitoes, Anopheles messeae were the most sensitive to AIFP, while Cx. modestus exhibited the least sensitivity to this formulation. Among nontarget aquatic invertebrates, the greatest toxicity of AIFP was observed for benthic species (larval Chironomus sp. Meigen (Diptera: Chironomidae), whereas predators (dragonflies, water beetles, and water bugs) exhibited the lowest AIFP sensitivity. AIFP sensitivity of the clam shrimp Lynceus brachyurus O. F. Muller (Diplostraca: Lynceidae), the phantom midge Chaoborus crystallinus De Geer (Diptera: Chaoboridae), and the mayfly Caenis robusta Eaton (Ephemeroptera: Caenidae) was intermediate and similar to the sensitivity of the mosquito Cx. modestus. However, these nontarget species were more resistant than An. messeae and Ae. aegypti. Solid-phase extraction of mosquito larvae treated with AIFP and subsequent high-performance liquid chromatography (HPLC) analysis of the extracts revealed an AVM concentration of up to 2.1 ± 0.3 µg/g. Feeding the creeping water bug Ilyocoris cimicoides L. (Hemiptera: Naucoridae) on the AIFP-treated mosquito larvae resulted in 51% mortality of the predaceous species. But no toxicity was observed for Aeshna mixta Latreille (Odonata: Aeshnidae) dragonfly larvae fed those mosquito larvae. The results of this work showed that this AVM formulation can be effective against mosquito larvae.

The use of an in vitro approach to assess marine invertebrate carboxylesterase responses to chemicals of environmental concern.

Carboxylesterases (CEs) are key enzymes which catalyse the hydrolysis reactions of multiple xenobiotics and endogenous ester moieties. Given their growing interest in the context of marine pollution and biomonitoring, this study focused on the in vitro sensitivity of marine invertebrate CEs to some pesticides, pharmaceuticals, personal care products and plastic additives to assess their potential interaction on this enzymatic system and its suitability as biomarkers. Three bivalves, one gastropod and two crustaceans were used and CEs were quantified following current protocols set for mammalian models. Four substrates were screened for CEs determination and to test their adequacy in the hepatic fraction measures of the selected invertebrates. Two commercial recombinant human isoforms (hCE1 and hCE2) were also included for methodological validation. Among the invertebrates, mussels were revealed as the most sensitive to xenobiotic exposures while gastropods were the least as well as with particular substrate-specific preferences. Among chemicals of environmental concern, the plastic additive tetrabromobisphenol A displayed the highest CE-inhibitory capacity in all species. Since plastic additives easily breakdown from the polymer and may accumulate and metabolise in marine biota, their interaction with the CE key metabolic/detoxification processes may have consequences in invertebrate's physiology, affect bioaccumulation and therefore trophic web transfer and, ultimately, human health as shellfish consumers.