International audience | Triazoles belong to a family of fungicides that are ubiquitous in agroecosystems due to their widespread use in crops. Despite their efficiency in controlling fungal diseases, triazoles are also suspected to affect non-target vertebrate species through the disruption of key physiological mechanisms. Most studies so far have focused on aquatic animal models, and the potential impact of triazoles on terrestrial vertebrates has been overlooked despite their relevance as sentinel species of contaminated agroecosystems. Here, we examined the impact of tebuconazole on the thyroid endocrine axis, associated phenotypic traits (plumage quality and body condition) and sperm quality in wild-caught house sparrows (Passer domesticus). We experimentally exposed house sparrows to realistic concentrations of tebuconazole under controlled conditions and tested the impact of this exposure on the levels of thyroid hormones (T3 and T4), feather quality (size and density), body condition and sperm morphology. We found that exposure to tebuconazole caused a significant decrease in T4 levels, suggesting that this azole affects the thyroid endocrine axis, although T3 levels did not differ between control and exposed sparrows. Importantly, we also found that exposed females had an altered plumage structure (larger but less dense feathers) relative to control females. The impact of tebuconazole on body condition was dependent on the duration of exposure and the sex of individuals. Finally, we did not show any effect of exposure to tebuconazole on sperm morphology. Our study demonstrates for the first time that exposure to tebuconazole can alter the thyroid axis of wild birds, impact their plumage quality and potentially affect their body condition. Further endocrine and transcriptomic studies are now needed not only to understand the underlying mechanistic effects of tebuconazole on these variables, but also to further investigate their ultimate consequences on performance (i.e. reproduction and survival).

Numerous toxicological studies have shown that ingestion of pesticides can induce physiological stress in breeding birds, with adverse consequences on egg laying parameters and offspring quality through parental effects. However, previous studies do not mimic current levels of pesticide residues in typical landscapes, and they do not consider potential cocktail effects of pesticides as they occur in the wild. Herein, we explored whether realistic pesticide exposure affected reproduction parameters and offspring condition through parental effects in Grey partridge. We fed 24 breeding pairs with either seeds from conventional agriculture crops treated with various pesticides during cropping, or organic grains without pesticide residues as controls. The conventional and organic grain diets mimicked food options potentially encountered by wild birds in the field. The results showed that ingesting low pesticide doses over a long period had consequences on reproduction and offspring quality without altering mortality in parents or chicks. Compared with organic pairs, conventional pairs yielded smaller chicks at hatching that had a lower body mass index at 24 days old. Additionally, these chicks displayed lower haematocrit when body mass index was higher. Therefore, ingestion of conventional grains by parents resulted in chronic exposure to pesticide residues, even at low doses, and this had detrimental consequences on offspring. These results demonstrate a sublethal effect of pesticide residues through parental effects. The consequences of parental exposure on chicks might partly explain the decline in wild Grey partridge populations, which raises questions for avian conservation and demography if current agrosystem approaches are continued.

Anthropogenic pollution is a major driver of global environmental change. To be properly addressed, the study of the impact of pollutants must consider both lethal effects and sublethal effects on individual fitness. However, measuring fitness remains challenging. In plants, the total number of seeds produced, i.e. the seed set, is traditionally considered, but is not readily accessible. Instead, performance traits related to survival, e.g., vegetative biomass and reproductive success, can be measured, but their correlation with seed set has rarely been investigated. To develop accurate estimates of seed set, relationships among 15 vegetative and reproductive traits were analyzed. For this purpose, Noccaea caerulescens (Brassicaceae), a model plant to study local adaptation to metal-contaminated environments, was used. To investigate putative variation in trait relationships, sampling included several accessions cultivated in contrasting experimental conditions. To test their applicability, selected estimates were used in the first generation of a Laboratory Natural Selection (LNS) experiment exposing experimentally plants to zinc soil pollution. Principal component analyses revealed statistical independence between vegetative and reproductive traits. Traits showing the strongest positive correlation with seed set were the number of non-aborted silicles, and the product of this number and mean silicle length. They thus appeared the most appropriate to document sublethal or fitness effects of environmental contaminants in plant ecotoxicological studies. The relevance of both estimates was confirmed by using them to assess the fitness of parental plants of the first generation of an LNS experiment: the same families consistently displayed the highest or the lowest performance values in two independent experimental metal-exposed populations. Thus, both these fitness estimates could be used to determine the expected number of offspring and the composition of successive generations in further LNS experiments investigating the impact of multi-generational exposure of a plant species to environmental pollution.

The mosquito Aedes aegypti is a primary vector for major arboviruses, and its control is mainly based on the use of insecticides. Caffeine and spent coffee grounds (CG) are potential agents in controlling Ae. aegypti by reducing survival and blocking larval development. In this study, we analyzed the effects of treatment with common CG (CCG: with caffeine), decaffeinated CG (DCG: with low caffeine), and pure caffeine on the survival, behavior, and morphology of the midgut of Ae. aegypti under laboratory conditions. Third instar larvae (L3) were exposed to different concentrations of CCG, DCG, and caffeine. All compounds significantly affected larval survival, and sublethal concentrations reduced larval locomotor activity, delayed development, and reduced adult life span. Damage to the midgut of treated larvae included changes in epithelial morphology, increased number of peroxidase-positive cells (more abundant in DCG-treated larvae), and caspase 3-positive cells (more abundant in CCG-treated larvae), suggesting that the treatments triggered cell damage, leading to activation of cell death. In addition, the treatments reduced the FMRFamide-positive enteroendocrine cells and dividing cells compared to the control. CG and caffeine have larvicidal effects on Ae. aegypti that warrant field testing for their potential to control mosquitoes.

Despite intensive ecotoxicological research, we still know relatively little about the ecological impacts of many environmental contaminants. Filling these knowledge gaps is particularly important regarding amphibians, because they play significant roles in freshwater and terrestrial ecosystems, and their populations are declining worldwide. In this study, we investigated two pollutants that have been poorly studied in ecotoxicology despite their widespread occurrence in surface waters: the herbicide terbuthylazine and the pharmaceutical drug carbamazepine. We exposed two anuran species throughout their larval development to each of two environmentally relevant concentrations of each pollutant, and recorded mortality and 17 sub-lethal endpoints up to several months after exposure. Mortality was low and unrelated to treatment. In agile frogs (Rana dalmatina), we found that treatment with 0.3 μg/L terbuthylazine decreased tadpole activity and reduced fat bodies in juveniles, whereas treatment with 50 μg/L carbamazepine decreased spleen size and increased spleen pigmentation. In common toads (Bufo bufo), treatment with 0.003 μg/L terbuthylazine increased body mass at metamorphosis, treatment with 0.3 μg/L terbuthylazine increased the size of optic tecta, and treatment with 0.5 μg/L carbamazepine decreased hypothalamus size. Treatment with 50 μg/L carbamazepine reduced the feeding activity of toad tadpoles, decreased their production of anti-predatory bufadienolide toxins, and increased their body mass at metamorphosis; juvenile toads in this treatment group had reduced spleen pigmentation. Neither treatments affected the time to metamorphosis, post-metamorphic body mass, or sex ratios significantly. These results show that environmental levels of both terbuthylazine and carbamazepine can have several sub-lethal effects on anurans, which may be detrimental to individual fitness and population persistence in natural conditions. Our findings further highlight that toxic effects cannot be generalized between chemicals of similar structure, because the terbuthylazine effects we found do not conform with previously reported effects of atrazine, a related and extensively studied herbicide.

The use of some systemic insecticides has been banned in Europe because they are toxic to beneficial insects when these feed on nectar. A recent study shows that systemic insecticides can also kill beneficial insects when they feed on honeydew. Honeydew is the sugar-rich excretion of hemipterans and is the most abundant carbohydrate source for beneficial insects such as pollinators and biological control agents in agroecosystems. Here, we investigated whether the toxicity of contaminated honeydew depends on i) the hemipteran species that excretes the honeydew; ii) the active ingredient, and iii) the beneficial insect that feeds on it. HPLC-MS/MS analyses demonstrated that the systemic insecticides pymetrozine and flonicamid, which are commonly used in Integrated Pest Management programs, were present in honeydew excreted by the mealybug Planococcus citri. However, only pymetrozine was detected in honeydew excreted by the whitefly Aleurothixus floccosus. Toxicological studies demonstrated that honeydew excreted by mealybugs feeding on trees treated either with flonicamid or pymetrozine increased the mortality of the hoverfly Sphaerophoria rueppellii, but did not affect the parasitic wasp Anagyrusvladimiri. Honeydew contaminated with flonicamid was more toxic for the hoverfly than that contaminated with pymetrozine. Collectively, our data demonstrate that systemic insecticides commonly used in IPM programs can contaminate honeydew and kill beneficial insects that feed on it, with their toxicity being dependent on the active ingredient and hemipteran species that excretes the honeydew.

The Paraná River basin is one of the most important in South America and is affected by human activities that take place on its margins. In particular, the De la Cruz stream flows through an industrial pole and the Arrecifes River goes mainly through agricultural fields. The aim of this study was to evaluate the water quality of the De la Cruz stream (S1) and the Arrecifes River (S2) by means of physicochemical parameters, including metals and pesticides concentrations. Since amphibians are good indicators of environmental quality, bioassays with Rhinella arenarum were carried on. For lethal and sublethal parameters, embryos and larvae were exposed to a dilution gradient of water samples and AMPHITOX Solution (AS) as negative control for 504 h. For the determination of oxidative stress biomarkers (Catalase -CAT-, Glutathione S-Transferase -GST-, Reduced Glutathione -GSH-, and lipid peroxidation -TBARS-), embryos and larvae were exposed to undiluted water samples and AS. For the determination of micronuclei, larvae at hind limb bud stage (S.28) were exposed to undiluted water samples, simultaneously with negative and positive controls (AS and cyclophosphamide 40 mg/L, respectively). Dissolved oxygen was low in both sites and the copper levels exceeded the Argentine limit for the protection of aquatic life. In embryos exposure, water sample from S1 caused lethal effects (504h-LC50 = 49 (28–71.6)%), increased TBARS levels, and GST and CAT activities. In larvae exposure, water sample from this site decreased CAT activity, while the water sample from S2 caused important lethal effects (504h-LC50 = 98.72 (60.60–302.52)%), low GSH levels and increased GST activity. Water samples from both sites induced higher micronuclei frequency than the negative control. This study alerts about the degradation of water quality of the studied sites including lethal and sublethal effects in R. arenarum that can jeopardize the native populations of this species.

Due to increased use of agrochemicals and growing concerns about ecotoxicology, the development of new insecticides, moving away from those with neurotoxic and broad spectrum effects towards insecticides that are safer for the environment and nontarget beneficial species, has been a research priority. Novaluron stands out among these newer insecticides, is an insect growth regulator that is used for the control of insect pests in crops grown close to mulberry plantations. Mulberry serves as food for the silkworm Bombyx mori, which is a nontarget insect of great economic importance to silk production. We investigated the lethal and sublethal effects of Novaluron on the development of B. mori. Larvae were segregated into experimental groups: the control groups (CGs) and the treatment groups (TGs), which were treated with the Novaluron concentration of 0.15 mL/L. Following exposure, we analyzed: larval mortality, changes in the insect life cicle and cytotoxic effects on the midgut cells. This is the first report about the Novaluron’s effects on B.mori. We detected rupture in the integument, complete cessation of feeding, late development, incomplete ecdysis and production of defective cocoons. After 240 h of exposure, there was 100% mortality in TG larvae exposed in the 3rd instar and 20% mortality from larvae exposed in the 5th instar. Cytotoxic effects was observed, such as dilation of cells, emission of cytoplasmic protrusions, extreme rarefaction of the cytoplasm and nuclei, dilation of the endoplasmic reticulum in addition to changes in mitochondria, the presence of large digestive vacuoles and intercellular spaces and the presence of active caspase. Novaluron exposure impairs the midgut and may affect the physiological functions of this organ. Novaluron additionally compromises several phases of insect development, indicating the importance of toxicology studies that utilize different life stages of nontarget species to evaluate the safe use of insecticides.

Leaf-cutting ants are important pests of forests and agricultural crops in the Neotropical region. Atta sexdens colonies can be composed of thousands of individuals, which form a highly complex society with a single reproductive queen. Successful control of this species is achieved only if the queen is affected. Few data are available on the lethal or sublethal effects of toxic compounds on leaf-cutting ant queens. Azadirachtin has been claimed as an effective biopesticide for insect control, but its action on leaf-cutting ants has been little explored. This study shows that azadirachtin affects oviposition in A. sexdens queens, impairing egg development by decreasing protein reserves. Azadirachtin inhibits the synthesis of vitellogenin, the major yolk protein precursor. The negative effects of azadirachtin on the reproduction of leaf-cutting ant queens suggest a potential use for the control of these insects.

Harmful microalgal blooms are a threat to aquatic organisms, ecosystems and human health. Toxic dinoflagellates of the genus Alexandrium are known to produce paralytic shellfish toxins and to release bioactive extracellular compounds (BECs) with potent cytotoxic, hemolytic, ichtyotoxic and allelopathic activity. Negative allelochemical interactions refer to the chemicals that are released by the genus Alexandrium and that induce adverse effects on the physiology of co-occurring protists and predators. Releasing BECs gives the donor a competitive advantage that may help to form dense toxic blooms of phytoplankton. However BECs released by Alexandrium minutum are uncharacterized and it is impossible to quantify them using classical chemical methods. Allelochemical interactions are usually quantified through population growth inhibition or lytic-activity based bioassays using a secondary target organism. However these bioassays require time (for growth or microalgal counts) and/or are based on lethal effects. The use of pulse amplitude modulation (PAM) fluorometry has been widely used to assess the impact of environmental stressors on phytoplankton but rarely for allelochemical interactions. Here we evaluated the use of PAM and propose a rapid chlorophyll fluorescence based bioassay to quantify allelochemical BECs released from Alexandrium minutum. We used the ubiquitous diatom Chaetoceros muelleri as a target species. The bioassay, based on sub-lethal effects, quantifies allelochemical activity from different samples (filtrates, extracts in seawater) within a short period of time (2 h). This rapid bioassay will help investigate the role of allelochemical interactions in Alexandrium bloom establishment. It will also further our understanding of the potential relationship between allelochemical activities and other cytotoxic activities from BECs. While this bioassay was developed for the species A. minutum, it may be applicable to other species producing allelochemicals and may provide further insights into the role and impact of allelochemical interactions in forming dense algal blooms and structuring marine ecosystems.