Sub-lethal effects of the insecticide, imidacloprid, on the responses of damselfly larvae to chemosensory cues indicating predation risk.

Insecticides, including the widely used neonicotinoids, can affect both pest and non-target species. In addition to lethal effects, these insecticides at sub-lethal levels may cause disruption to sensory perception and processing leading to behavioural impairments. In this laboratory experiment, we investigated the effects of a 10-day exposure to the neonicotinoid insecticide, imidacloprid, on the behaviour of larvae of the damselfly, Lestes congener. In tests of baseline activity, imidacloprid concentrations of 1.0 and 10.0 µg/L caused significant reductions in foraging behaviour. Moreover, in response to chemical cues that indicate a potential risk to the larvae, imidacloprid caused the loss of an appropriate antipredator response (reduced foraging) depending on the concentration and duration of exposure. Imidacloprid at 0.1 µg/L caused the loss of responses toward the odour of a beetle (Dytiscus spp.) predator after 10 days of exposure, whereas 1.0 µg/L caused lost responses toward both the predator odour and injured conspecific cues (i.e., alarm cues) and after only 2 days of exposure. However, at 10.0 µg/L, larvae responded appropriately to both cues throughout the duration of the study, suggesting compensatory responses to imidacloprid at higher concentrations. Hence, the lack of appropriate responses at 1.0 µg/L likely resulted from a cognitive impairment rather than chemical alteration of these important chemosensory cues. In the natural environment, such effects will likely cause decreased survivorship in predator encounters. Hence, imidacloprid exposure, even at low concentrations, could have adverse consequences for chemosensory ecology of this damselfly species.

Combinations of Lemongrass and Star Anise Essential Oils and Their Main Constituent: Synergistic Housefly Repellency and Safety against Non-Target Organisms.

The present study evaluated the housefly repellency of single-component formulations and combinations of lemongrass and star anise essential oils (EOs) and their main constituents. The efficacies of the combinations were compared against those of single-component formulations and DEET. Safety bioassays of all formulations and DEET on non-target species-guppy, molly, dwarf honeybee, and stingless bee-were conducted. GC-MS analysis showed that the main constituent of lemongrass EO was geranial (46.83%) and that of star anise EO was trans-anethole (92.88%). All combinations were highly synergistic compared to single-component formulations, with an increased repellent value (IR) of 34.6 to 51.2%. The greatest synergistic effect was achieved by 1.0% lemongrass EO + 1.0% trans-anethole combination, with an IR of 51.2%. The strongest, 100% repellent rate at 6 h was achieved by 1.0% geranial + 1.0% trans-anethole. They were twice as effective as DEET and caused obvious damage to housefly antennae under microscopic observation. All single-component formulations and combinations were benign to the four tested non-target species. In contrast, DEET was highly toxic to them. The synergistic repellency and biosafety of these two combinations are compellingly strong support for developing them into an effective green repellent.

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.

Adulticidal synergy of two plant essential oils and their major constituents against the housefly Musca domestica and bioassay on non-target species.

Single and mixture formulations of lemongrass (Cymbopogon citratus (DC.) Stapf.) and star anise (Illicium verum (J. Presl.)) essential oils (EOs) and their major constituents were assayed for their adulticidal activities against housefly, Musca domestica L., and two non-target species, stingless bee (Tetragonula pegdeni Schwarz) and guppy (Poecilia reticulata Peters). The efficacies of the mixture formulations were compared against those of the single formulations and 1.0% α-cypermethrin, a common synthetic insecticide. GC-MS analysis found that the major constituent of lemongrass EO was geranial (45.23%), and that of star anise EO was trans-anethole (93.23%). Almost all mixture formulations were more effective in adulticidal activity against housefly adults than single formulations and 1.0% α-cypermethrin. A mixture of 1.0% lemongrass EO + 1.0% trans-anethole exhibited the strongest synergistic insecticidal activity with a 100% mortality rate (KT50 of 3.2 min and LT50 of 0.07 h). The relative percentage increase in mortality rate over single formulations was between 1.6 and 91.9%. In addition, it was three times more effective than 1.0% α-cypermethrin. To find the mechanism of adulticidal action, scanning electron microscopy (SEM) was done to find morphological aberrations, such as antennal and mouthpart aberrations, after the houseflies were treated with 1.0% lemongrass EO + 1.0% trans-anethole. The aberrations included deformed and abnormal shape of arista and flagellum, change in labellum pigmentation, and damage to pseudotracheae. Regarding toxicity against non-target species, all single and mixture formulations were not toxic to the two non-target species, while 1.0% α-cypermethrin was highly toxic. To conclude, a mixture of 1.0% lemongrass EO + 1.0% trans-anethole can be an excellent, natural, sustainable housefly adulticidal agent.

Exposure to the insecticide, imidacloprid, impairs predator-recognition learning in damselfly larvae.

Many aquatic organisms use chemosensory information to learn about local predation threats, but contaminants in their environment may impair such cognitive processes. Neonicotinoids are a class of water-soluble systemic insecticides that have become a major concern in aquatic systems. In this study, we explored how a 10-day exposure to various concentrations (0, 0.1, 1.0, or 10.0 µg/L) of the neonicotinoid imidacloprid affects the learned recognition of predator odour by non-target damselfly larvae (Lestes spp). Unexposed larvae and those exposed to the low concentration (0.1 µg/L) demonstrated an appropriate learned response to a novel predator odour following a conditioning with the odour paired with chemical alarm cues. However, such learning failed to occur for larvae that were exposed to imidacloprid concentrations of 1.0 and 10.0 µg/L. Thus, either the cognitive processing of the chemical information was impaired or the chemistry of one or both of the conditioning cues was altered, making them ineffective for learning. In a second experiment, we found evidence for this latter hypothesis. In the absence of background imidacloprid exposure, larvae did not show significant learned responses to the predator odour when the conditioning cues were mixed with imidacloprid (initial pulse solution of 3.0 µg/L) at the start of conditioning (reaching a final concentration of 0.01 µg/L). These findings indicate that even low levels of imidacloprid can have important implications for chemosensory cognition of non-target species in aquatic environments.

Living on the edge: Predicting songbird response to management and environmental changes across an ecotone.

Effective wildlife management requires robust information regarding population status, habitat requirements, and likely responses to changing resource conditions. Single-species management may inadequately conserve communities and result in undesired effects to non-target species. Thus, management can benefit from understanding habitat relationships for multiple species. Pinyon pine and juniper (Pinus spp. and Juniperus spp.) are expanding into sagebrush-dominated (Artemisia spp.) ecosystems within North America and mechanical removal of these trees is frequently conducted to restore sagebrush ecosystems and recover Greater Sage-grouse (Centrocercus urophasianus). However, pinyon-juniper removal effects on non-target species are poorly understood, and changing pinyon-juniper woodland dynamics, climate, and anthropogenic development may obscure conservation priorities. To better predict responses to changing resource conditions, evaluate non-target effects of pinyon-juniper removal, prioritize species for conservation, and inform species recovery within pinyon-juniper and sagebrush ecosystems, we modeled population trends and density-habitat relationships for four sagebrush-associated, four pinyon-juniper-associated, and three generalist songbird species with respect to these ecosystems. We fit hierarchical population models to point count data collected throughout the western United States from 2008 to 2020. We found regional population changes for 10 of 11 species investigated; 6 of which increased in the highest elevation region of our study. Our models indicate pinyon-juniper removal will benefit Brewer's Sparrow (Spizella breweri), Green-tailed Towhee (Pipilo chlorurus), and Sage Thrasher (Oreoscoptes montanus) densities. Conversely, we predict largest negative effects of pinyon-juniper removal for species occupying early successional pinyon-juniper woodlands: Bewick's Wren (Thryomanes bewickii), Black-throated Gray Warblers (Setophaga nigrescens), Gray Flycatcher (Empidonax wrightii), and Juniper Titmouse (Baeolophus ridgwayi). Our results highlight the importance of considering effects to non-target species before implementing large-scale habitat manipulations. Our modeling framework can help prioritize species and regions for conservation action, infer effects of management interventions and a changing environment on wildlife, and help land managers balance habitat requirements across ecosystems.

Evaluation of the Morphological, Physiological and Biochemical Effects Induced by Coragen 20 SC in Some Non-Target Species.

Coragen 20 SC is an insecticide based on chlorantraniliprole that is applied on many crops. Considered an effective product with an incremental cost-benefit ratio, it has been widely used globally. Residual pesticides affect non-target organisms, so it is necessary to explore the possible effects induced by these xenobiotics on different species. This work aimed to assess some morphological, physiological and biochemical effects induced by Coragen 20 SC on two non-target species: Perca fluviatilis (Linné, 1758) and Triticum aestivum L. The concentrations used were the same for all tested species (0.0125, 0.025 and 0.05 mL L-1), and the experiments were of the acute, subchronic and chronic type. The toxicological effects of Coragen 20 SC on perch recorded behavioral changes, a decrease in respiratory rate and oxygen consumption, an increase in blood glucose levels and a decrease in the number of erythrocytes and leukocytes. The results obtained from the evaluation of Coragen 20 SC toxicity using the Triticum test indicate a weak to moderate phytotoxicity for the considered parameters at the applied doses. Only the assimilatory pigments were significantly modified at the concentration of 0.025 mL L-1 for the growth of the axial organs and the wet and dry weight, with the changes obtained not being statistically significant.

A systematic review of the toxic effects of a nanopesticide on non-target organisms: Estimation of protective concentrations using a species sensitivity distribution (SSD) approach - The case of atrazine.

Nanopesticides, such as nanoencapsulated atrazine (nATZ), have been studied and developed as eco-friendly alternatives to control weeds in fields, requiring lower doses. This review contains a historical and systematic literature review about the toxicity of nATZ to non-target species. In addition, the study establishes protective concentrations for non-target organisms through a species sensitivity distribution (SSD) approach. Through the systematic search, we identified 3197 publications. Of these, 14 studies addressed "(nano)atrazine's toxicity to non-target organisms". Chronological and geographic data on the publication of articles, characterization of nATZ (type of nanocarrier, size, polydispersity index, zeta potential), experimental design (test species, exposure time, measurements, methodology, tested concentrations), and toxic effects are summarized and discussed. The data indicate that cell and algal models do not show sensitivity to nATZ, while many terrestrial and aquatic invertebrates, aquatic vertebrates, microorganisms, and plants have high sensitivity to nAZT. The SSD results indicated that D. similis is the most sensitive species to nATZ, followed by C. elegans, E. crypticus, and P. subcapitata. However, the limitations in terms of the number of species and endpoints available to elaborate the SSD reflect gaps in knowledge of the effects of nATZ on different ecosystems.

Neonicotinoid exposure in Tricolored Blackbirds (Agelaius tricolor).

There is increasing awareness of the negative ecological and environmental effects of widespread use of pesticides on the landscape. Spillover or drift of pesticides from agricultural areas has been shown to impact species health, reproduction, and trophic dynamics through both direct and indirect mechanisms. Neonicotinoid insecticides are associated with observed declines of insectivorous and grassland birds, and these environmental pollutants are a significant conservation concern for many species that have experienced past or current population declines. Due to the high efficacy of these modern insecticides in depressing local insect populations, insectivorous birds can be negatively impacted by a pesticide-mediated reduction in food supply. Neonicotinoids may act synergistically with other stressors, such as habitat loss, to exacerbate threats to species or population viability. The Tricolored Blackbird is an insectivorous grassland bird of conservation concern in California, USA. Due to the high association of this species with agricultural habitats, we sought to quantify the amount of neonicotinoid residues in Tricolored Blackbird carcasses as a first step in assessing how this species may be impacted by pesticides. Out of 85 salvaged carcasses sampled (N = 24 adults, N = 3 fledglings, and N = 58 nestlings), only two contained detectable levels of target compounds. These were an adult and one nestling that contained clothianidin residue (40 ppb and 7 ppb, respectively); both of these birds were salvaged from breeding colonies associated with dairy farms in Kern County, California. We suggest that further work is needed to assess neonicotinoid exposure of Tricolored Blackbirds in dairy-associated breeding colonies.

Ecotoxicological relevance of glyphosate and flazasulfuron to soil habitat and retention functions - Single vs combined exposures.

Glyphosate (GLY) and flazasulfuron (FLA) are two non-selective herbicides commonly applied together. However, research focused on their single and combined ecotoxicological impacts towards non-target organisms is still inconclusive. Therefore, this study aimed to test their single effects on soil's habitat and retention functions, and to unravel their combined impacts to earthworms and terrestrial plants. For this, ecotoxicological assays were performed with plants (Medicago sativa), oligochaetes (Eisenia fetida) and collembola (Folsomia candida). Soil elutriates were also prepared and tested in macrophytes (Lemna minor) and microalgae (Raphidocelis subcapitata). FLA (82-413 µg kg-1) reduced earthworms' and collembola's reproduction and severely impaired M. sativa growth, being much more toxic than GLY (up to 30 mg kg-1). In fact, the latter only affected plant growth (≥ 9 mg kg-1) and earthworms (≥ 13 mg kg-1), especially at high concentrations, with no effects on collembola. Moreover, only elutriates from FLA-contaminated soils significantly impacted L. minor and R. sucapitata. The experiments revealed that the co-exposure to GLY and FLA enhanced the toxic effects of contaminated soils not only on plants but also on earthworms'. However, such increase in toxicity was dependent on GLY residual concentrations in soils. Overall, this work underpins that herbicides risk assessment should consider herbicides co-exposures, since the evaluation of single exposures is not representative of current phytosanitary practices and of the potential effects under field conditions, where residues of different compounds may persist in soils.

Habitat and Season Effects on Small Mammal Bycatch in Live Trapping.

Trapping small mammals is frequently used to study the dynamics, demography, behavior and presence of pathogens. When only particular small mammal species are in the focus of interest, all other species are unnecessary bycatch. We analyzed data from extensive live trapping campaigns conducted over the last decade in Germany, following a consistent standard trapping protocol that resulted in about 18,500 captures of small mammals. Animals were trapped with Ugglan multiple capture traps in grassland, forest and margin habitat. Trap success and the proportion of bycatch were about 30% when target species were common voles (Microtus arvalis) in grassland and common voles and bank voles (Clethrionomys glareolus) in margins and forests. This was more pronounced in spring and along margins. Species mentioned in the early warning list according to the Red List Germany were higher in numbers and proportion in spring and in grassland. The results will help to avoid periods with enhanced presence of bycatch, including endangered species (if the purpose of the study allows) or to pay particular attention in certain seasons and habitats when the occurrence of bycatch is most likely.

Second-generation anticoagulant rodenticides in the blood of obligate and facultative European avian scavengers.

The widespread use of second-generation anticoagulant rodenticides (SGARs) and their high persistence in animal tissues has led to these compounds becoming ubiquitous in rodent-predator-scavenger food webs. Exposure to SGARs has usually been investigated in wildlife species found dead, and despite growing evidence of the potential risk of secondary poisoning of predators and scavengers, the current worldwide exposure of free-living scavenging birds to SGARs remains scarcely investigated. We present the first active monitoring of blood SGAR concentrations and prevalence in the four European obligate (i.e., vultures) and facultative (red and black kites) avian scavengers in NE Spain. We analysed 261 free-living birds and detected SGARs in 39.1% (n = 102) of individuals. Both SGAR prevalence and concentrations (ΣSGARs) were related to the age and foraging behaviour of the species studied. Black kites showed the highest prevalence (100%), followed by red kites (66.7%), Egyptian (64.2%), bearded (20.9%), griffon (16.9%) and cinereous (6.3%) vultures. Overall, both the prevalence and average ΣSGARs were higher in non-nestlings than nestlings, and in species such as kites and Egyptian vultures foraging in anthropic landscapes (e.g., landfill sites and livestock farms) and exploiting small/medium-sized carrions. Brodifacoum was most prevalent (28.8%), followed by difenacoum (16.1%), flocoumafen (12.3%) and bromadiolone (7.3%). In SGAR-positive birds, the ΣSGAR (mean ± SE) was 7.52 ± 0.95 ng mL-1; the highest level detected being 53.50 ng mL-1. The most abundant diastereomer forms were trans-bromadiolone and flocoumafen, and cis-brodifacoum and difenacoum, showing that lower impact formulations could reduce secondary exposures of non-target species. Our findings suggest that SGARs can bioaccumulate in scavenging birds, showing the potential risk to avian scavenging guilds in Europe and elsewhere. We highlight the need for further studies on the potential adverse effects associated with concentrations of SGARSs in the blood to better interpret active monitoring studies of free-living birds.

Transcriptome profiling of the early developmental stages in the giant mussel Choromytilus chorus exposed to delousing drugs.

The giant mussel Choromytilus chorus is a marine bivalve commonly collected in central - southern Chile from fishery zones shared with the salmon industry. These economically relevant areas are also affected by the use of pesticides for controlling sea lice infestations in salmon aquaculture. Their main target is the sea louse Caligus rogercresseyi. However, other than some physiological impacts, the molecular effects of delousing drugs in non-target species such as C. chorus remain largely understudied. This study aimed to explore the transcriptome modulation of Trochophore and D larvae stages of C. chorus after exposure to azamethiphos and deltamethrin drugs. Herein, RNA-seq analyses and mRNA-lncRNAs molecular interactions were obtained. The most significant changes were found between different larval development stages exposed to delousing drugs. Notably, significant transcriptional variations were correlated with the drug concentrations tested. The biological processes involved in the development, such as cell movement and transcriptional activity, were mainly affected. Long non-coding RNAs (lncRNAs) were also identified in this species, and the transcription activity showed similar patterns with coding mRNAs. Most of the significantly expressed lncRNAs were associated with genes annotated to matrix metalloproteinases, collagenases, and transcription factors. This study suggests that exposure to azamethiphos or deltamethrin drugs can modulate the transcriptome signatures related to the early development of the giant mussel C. chorus.

Asymptomatic Anticoagulant Rodenticide Exposure in Dogs and Cats-A French and Belgian Rural and Urban Areas Study.

Anticoagulant rodenticides (ARs) are important tools for controlling rodent pests, but they also pose a health threat to non-target species. ARs are one of the most common causes of pet poisoning. However, exposure of domestic animals to subclinical doses of ARs is poorly documented. To study the random exposure of dogs and cats to ARs, feces from animals showing no clinical signs of rodenticide poisoning were collected from a network of French and Belgian veterinarians. We analyzed fresh feces from 304 dogs and 289 cats by liquid chromatography-tandem mass spectrometry. This study showed a limited prevalence of AR exposure in dogs and cats of 2.6 and 4.5% respectively. In both species, access to the outdoors is a risk factor for ARs exposure. In contrast, the sex of the animals did not affect the ARs exposure status. The observation of the ratio of cis and trans isomers suggested primary exposure in dogs, but also in some cats. While primary exposure in dogs appears to be related to the use of ARs as plant protection products, primary exposure in cats may be malicious, as warfarin, an anticoagulant formerly used as a rodenticide and now used only in humans, was found in 4 of 13 exposed cats. Secondary exposure may also occur in cats.Our study showed reduced exposure in dogs and cats, compared to wildlife, which often has high exposure, especially in areas where rodent control is important.

Impacts on antioxidative enzymes and transcripts in darter (Etheostoma spp.) brains in the Grand River exposed to wastewater effluent.

The Grand River watershed is the largest in southern Ontario and assimilates thirty wastewater treatment plants (WWTP) with varied degrees of treatment. Many WWTPs are unable to effectively eliminate several contaminants of emerging concern (CECs) from final effluent, leading to measurable concentrations in surface waters. Exposures to CECs have reported impacts on oxidative stress measured through antioxidative enzymes (SOD, CAT, GPX). This study focuses on the effects of WWTP effluent on four Etheostoma (Darter) species endemic to the Grand River, by investigating if increased antioxidative response markers are present in darter brains downstream from the effluent outfall compared to an upstream reference site relative to the Waterloo, ON WWTP across two separate years (Oct 2020 and Oct 2021). This was assessed using transcriptional and enzyme analysis of antioxidant enzymes and an enzyme involved in serotonin synthesis, tryptophan hydroxylase (tph). In fall 2020, significant differences in transcript markers were found between sites and sexes in GSD with SOD and CAT showing increased expression downstream, in JD with both sexes showing increased SOD downstream, and an interactive effect for tph in RBD. Changes in transcripts aligned with enzyme activity where interactive effects with sex-related differences were observed in fish collected fall 2020. In contrast, transcripts measured in fall 2021 were increased upstream compared to downstream species in RBD and GSD. This study additionally displayed yearly, species and sex differences in antioxidant responses. Continued investigation on the impacts of CECs in effluent in non-target species is required to better understand WWTP effluent impacts.

Effect of genetically modified maize expressing the Cry1Ab and EPSPS proteins on growth, development, and gut bacterial diversity of the non-target arthropod Locusta migratoria.

The widespread cultivation of genetically modified (GM) crops has raised concerns for their safety. Here, we evaluated the effects of a GM maize variety expressing the Cry1Ab (14.76 ± 0.87 µg/g FW) and EPSPS proteins (191.55 ± 15.69 µg/g FW) on the life-history traits and gut bacterial community of a non-target arthropod, Locusta migratoria, in the laboratory. We found that GM maize had no significant effect on the survival or body weight of different development stages of L. migratoria. The midgut and hindgut bacterial diversities and compositions were determined using high-throughput sequencing targeting the V3-V4 regions of the 16S rRNA. No significant changes were found in the species diversity or abundance between insects in the GM-fed treatment and the non-GM control. Furthermore, the concentration of Cry1Ab and EPSPS in the gut was determined after digestion of GM maize. Results showed that the contents of Cry1Ab/EPSPS rapidly decreased and were hard to detect after 72 h. Based on the parameters assessed, we can conclude that the GM maize variety examined has no significant adverse effect on L. migratoria.

Veterinary pharmaceuticals as a threat to endangered taxa: Mitigation action for vulture conservation.

Overuse and misapplication of veterinary pharmaceuticals affect the ecosystem, even at low concentrations. Vultures are mainly exposed to these compounds when feeding on improperly disposed carcasses from animals treated before death. This produces diverse negative impacts on vulture health and populations, even leading to death. Using the available bibliography we determined which veterinary pharmaceuticals vultures are exposed to worldwide and assessed the potential consequences for these species. Based on the responsibilities of the different stakeholders, we also propose action to mitigate this problem. Of 104 articles addressing vulture exposure to veterinary pharmaceuticals, most came from Asia, Europe and Africa; almost no information was available on the Americas. Vultures were reported as being exposed to non-steroidal anti-inflammatory drugs (NSAIDs), antibiotics, anti-parasitic and euthanizing agents. Most available information is related to the catastrophic effect of the NSAID diclofenac in South Asia. Vultures are particularly exposed to veterinary drugs when ingesting carcasses from intensive livestock production, but other potential pathways (e.g., discards from salmon farms or fisheries) have not yet been properly evaluated. It is essential to improve scientific information on this topic - increasing the range of drugs and geographical areas studied - in order to implement sustainable conservation action for these birds. A combination of strategies could prove effective in reducing the impact of pharmaceuticals on the environment and non-target species. To mitigate this conservation problem, a set of multilateral actions should therefore be implemented, involving diverse stakeholders such as government representatives, pharmaceutical companies, veterinary practitioners, scientists and conservation agents, and local communities.

Antifungal and Insecticidal Potential of the Essential Oil from Ocimum sanctum L. against Dangerous Fungal and Insect Species and Its Safety for Non-Target Useful Soil Species Eisenia fetida (Savigny, 1826).

The antifungal and insecticidal effect of the essential oil from Ocimum sanctum L. was evaluated using a model set of harmful organisms hazardous for health and the economy. Toxigenic and plant pathogenic filamentous fungi, including causal agents of human infections, were chosen as exemplary fungal groups-Fusarium verticillioides, Penicillium expansum and Aspergillus flavus. Spodoptera littoralis (African cotton leafworm), Culex quinquefasciatus (Southern house mosquito), the lymphatic filariasis vector and potential Zika virus vector, and the common housefly, Musca domestica were chosen as model insects. Major and minor active substances were detected and quantified using GC/MS analysis. Environmental safety was verified using the non-target useful organism Eisenia fetida. Significant antifungal and insecticidal activity, as well as environmental safety, were confirmed. The essential oil showed the highest efficacy against A. flavus according to MIC50/90, and against S. littoralis larvae according to LD50/90. The monoterpenoid alcohol linalool, t-methyl cinnamate, and estragole as phenylpropanoids were detected as effective major components (85.4%). The essential oil from Ocimum sanctum L. was evaluated as universal and significantly efficient, providing a high potential for use in environmentally safe botanical 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.

Effects of diclofenac on the swimming behavior and antioxidant enzyme activities of the freshwater interstitial crustacean Bryocamptus pygmaeus (Crustacea, Harpacticoida).

Diclofenac (DCF) is one of the most widespread pharmaceutical compounds found in freshwaters as a pseudo-persistent pollutant due to its continuous release from point and diffuse sources, being its removal in Wastewater Treatment Plants incomplete. Moreover, DCF is particularly persistent in interstitial habitats and potentially toxic for the species that spend their whole life cycle among the same sediment grains. This study is aimed at offering a first contribution to the assessment of DCF effects on freshwater invertebrate species living in the interstitial habitats of springs, rivers, lakes and groundwaters. The Crustacea Copepoda are one of the main components of the freshwater interstitial communities, with the primacy taken by the worm-like and small-sized harpacticoids. A sub-lethal concentration of 50 µg L-1 DCF significantly affected six out of the eight behavior parameters of the burrower/interstitial crustacean harpacticoid Bryocamptus pygmaeus recorded by video tracking analysis. DCF exposure reduced swimming speed, swimming activity, exploration ability and thigmotaxis, and increased swimming path tortuosity. The biochemical approach revealed a reduced level of the mitochondrial superoxide dismutase 2 in individuals exposed to DCF. It could be explained by a decline in mitochondrial performance or by a reduced number of functional mitochondria. Since mitochondrial dysfunction may determine ATP reduction, it comes that less energy is produced for maintaining the cell functions of the DCF-exposed individuals. In addition, the increasing energy demand for the detoxification process further contributes to decrease the total energetic budget allocated for other physiological activities. These observations can explain the changes we have observed in the swimming behavior of the copepod B. pygmaeus.