The combined effects of microbial insecticides and sodium chloride on the development and emergence of Chironomus xanthus.

BACKGROUND: Freshwater organisms are facing increasing salinity levels, not only due to natural environmental processes, but also human activities, which can cause several physiological adaptations to osmotic stress. Additionally, these organisms might also have to deal with contamination by microbial insecticides. Our main goal was to use Chironomus xanthus to assess the chronic effects of increasing the salinity and commercial formulations of the microbial insecticides based on Bacillus thuringiensis subs. kurstaki (Btk) and Beauveria bassiana (Bb) as active ingredients, respectively. RESULTS: A significant interaction of growth was observed between the biopesticide based on Bb and NaCl on the larvae of C. xanthus. Single exposure to NaCl and each one of the formulations demonstrated deleterious impacts not only on larval development, but also on the emergence success and emergence time of this nontarget insect, with potential consequences for freshwater ecosystems due to cascading effects. CONCLUSION: The chronic effects induced by both bioinsecticides show that these formulations can have environmental impacts on nontarget freshwater insects. © 2023 Society of Chemical Industry.

What does not kill it makes it stronger! The tolerance of the F1 larvae of Chironomus xanthus to a neonicotinoid insecticide formulation.

Thiamethoxam (TMX) is a systemic neonicotinoid that acts as a partial agonist of the nicotinic acetylcholine receptors (nAChRs). However, target species have shown resistance to formulations based on such neonicotinoids, which can also be expected for non-target insects. This research aimed to study the effects of a formulation based on TMX [Cruiser® 350 FS (CRZ)] on the life traits of Chironomus xanthus filial generation (F1) and compare it with the parental generation (P). Environmentally relevant concentrations of CRZ significantly decreased larvae growth P generation , also slowing and decreasing their emergence. Larvae of the F1 generation were less sensitive than their parents, suggesting that the progeny were able to thrive and perform basic physiological functions better than the parental generation. Our results highlight that insect resistance to neonicotinoids may be associated with the better performance of the filial generation, which is related to the change in affinities of the active ingredient for the sub-units constituting the nAChRs subtypes of F1 organisms, inherited from P organisms that were able to survive and reproduce. Moreover, further studies using biochemical and omics tools should be performed to disentangle the specific changes occurring at the nAChRs throughout insect development.

The sexual reproduction of the nontarget planarian Girardia tigrina is affected by ecologically relevant concentrations of difenoconazole: new sensitive tools in ecotoxicology.

The fungicide difenoconazole, widely used to reduce the negative impacts of fungi diseases on areas with intensive farming, can reach freshwater systems causing deleterious effects on nontarget organisms. The acute and chronic toxicity of a commercial formulation containing 250 g L-1 of difenoconazole (Prisma®) as the active ingredient was assessed in the freshwater planarian Girardia tigrina. The endpoints evaluated were feeding rate, locomotion, regeneration, and sexual reproduction of planarians. The estimated 48 h LC50 of the commercial formulation on planarians expressed as the concentration of the active ingredient difenoconazole was 47.5 mg a.i.L-1. A significant decrease of locomotion (LOEC = 18.56 mg a.i.L-1), delayed regeneration (LOEC = 9.28 mg a.i.L-1), and sexual reproduction impairment, i.e., decreased fecundity and fertility rates (LOEC ≤ 1.16 mg a.i.L-1) were observed on planarians exposed to sublethal concentrations of the formulation. This study demonstrated the importance of using reproductive, physiological, and behavioral parameters as more sensitive and complementary tools to assess the deleterious effects induced by a commercial formulation of difenoconazole on a nontarget freshwater organism. The added value and importance of our research work, namely, the impairment of sexual reproduction of planarians, contributes to the development of useful tools for ecotoxicology and highlights the fact that those tools should be developed as guidelines for testing of chemicals. Our results showed that the use of reproductive parameters of Girardia tigrina would help to complement and achieve a better assessment of the risk posed by triazole fungicides to freshwater ecosystems.

Do bio-insecticides affect only insect species? Behavior, regeneration, and sexual reproduction of a non-target freshwater planarian.

Bio-insecticides have been increasingly used worldwide as ecofriendly alternatives to pesticides, but data on their effects in non-target freshwater organisms is still scarce and limited to insects. The aim of this study was to determine the lethal and sub-lethal effects of the bio-insecticides Bac Control (based on Bacillus thuringiensis kurstaki-Btk) and Boveril (based on Beauveria bassiana-Bb) on regeneration, behavioral, and reproductive endpoints of the freshwater planarian Girardia tigrina. The estimated LC50-48h were > 800 mg a.i./L for Btk and 60.74 mg a.i./L for Bb. In addition, exposure to Btk significantly decreased locomotion and feeding activities of planarians (lowest observed effect concentration (LOEC) of 12.5 mg a.i./L Btk) and fecundity rate (LOEC = 3.12 mg a.i./L Btk), whereas exposure to Bb significantly delayed regeneration (LOEC = 0.75 mg a.i./L Bb) and decreased fecundity rate (1.5 mg a.i./L Bb) of planarians. Thus, both bio-insecticides induced deleterious sub-lethal effects on a non-insect freshwater invertebrate species. However, only Bb-based formulation affected the survival, fecundity rate, and regeneration at concentrations below the maximum predicted environmental concentration (PEC = 247 mg/L). Thus, care should be taken when using such formulations as alternatives to chemical insecticides near aquatic ecosystems.

Lethal and sublethal effects of the saline stressor sodium chloride on Chironomus xanthus and Girardia tigrina.

Salinization in freshwaters is gradually increasing as a result of human activities and climatic changes. Higher salt content causes stress for freshwater organisms. Sodium chloride (NaCl) is among the most frequently occurring salts in freshwater ecosystems. The objective of the present study was to investigate the lethal and sublethal effects of NaCl on freshwater ecosystems, using as test organism the dipteran Chironomus xanthus and the planarian Girardia tigrina. Acute tests showed that C. xanthus was more sensitive (48-h LC50 (median lethal concentration) of 2.97 g NaCl L-1) than G. tigrina (48-h LC50 of 7.77 g NaCl L-1). C. xanthus larvae growth rate (larvae length and head capsule width) was significantly reduced under exposure to concentrations as low as 0.19 g L-1 NaCl and higher. A delay in the emergence time (EmT50) was also demonstrated for the same concentration. Sublethal NaCl effects in G. tigrina included feeding inhibition (LOEC (lowest observed effect concentration) of 0.4 g L-1), reduced locomotion (LOEC = 0.2 g L-1), and 24-48-h blastema regeneration (LOEC = 0.2 g L-1 and 0.1 g L-1, respectively). The results demonstrated the toxicity of NaCl to C. xanthus and G. tigrina including sublethal effects that can result in negative consequences for populations in natural freshwaters under salinization.

Toxicity of microbial insecticides toward the non-target freshwater insect Chironomus xanthus.

BACKGROUND: Commercial formulations based on Bacillus thuringiensis subs. kurstaki (Btk) and Beauveria bassiana (Bb) are commonly used microbial insecticides in Brazil and other tropical regions. However, and despite being considered environmentally friendly, their use generates concerns regarding possible adverse ecological effects in freshwater ecosystems. Here, we evaluate the effects of these bioinsecticides on the tropical aquatic dipteran Chironomus xanthus under laboratory conditions. RESULTS: After laboratory exposures to these compounds 48-h median lethal concentration (LC50 ) values of 1534 µg a.i./L for Btk and of 6.35 µg a.i./L for Bb were estimated. Chronic assays revealed different sublethal effects: Btk-based bioinsecticide exposure reduced C. xanthus growth [lowest observed effect concentration (LOEC) was 126 µg a.i./L for head width], decreased emergence rate (LOEC = 8 µg a.i./L) and increased immunological response (LOEC = 50 µg a.i./L) measured as total hemocyte count in larvae hemolymph. Exposure to low concentrations of Bb-based insecticide also reduced C. xanthus growth (LOEC = 0.07 µg a.i./L for larvae body length measurements), and emergence rate (LOEC = 0.28 µg a.i./L), despite no clear effects on the total hemocyte counts. CONCLUSION: Our results suggest that low concentrations of Btk and Bb bioinsecticides are toxic to C. xanthus. Given their widespread use and occurrence in tropical freshwater systems, research is needed to evaluate the potential effects of these compounds concerning natural freshwater insect communities and ecosystem functioning. © 2019 Society of Chemical Industry.

Exposure to Roundup® affects behaviour, head regeneration and reproduction of the freshwater planarian Girardia tigrina.

The demand of glyphosate-based herbicides including Roundup® is rising in the tropics due to increase occurence of glyphosate-resistant weeds that require higher herbicide application rates but also because of their use associated with genetically engineered, glyphosate-tolerant crops. Consequently, there is now an excessive use of glyphosate in agricultural areas with potential adverse effects also for the surrounding aquatic environments. This study aimed to determine the sensitivity of the freshwater planarian Girardia tigrina to acute and chronic exposures of Roundup®. Planarians were exposed to a range of lethal and sub-lethal concentrations of Roundup® to determine the median lethal concentration (LC50) concerning its active ingredient glyphosate and also effects on locomotor velocity (pLMV), feeding rate, regeneration, reproductive parameters and morphological abnormalities. Regeneration endpoints included length of blastema and time for photoreceptors and auricles regeneration after decapitation, while effects on reproduction were assessed measuring fecundity (number of deposited cocoons) and fertility (number of hatchlings) over five weeks of exposure to glyphosate. The estimated 48 h LC50 of was 35.94 mg glyphosate/L. Dose dependent effects were observed for feeding, locomotion and regeneration endpoints with Lowest observed effect concentration (LOEC) values as low as 3.75 mg glyphosate/L. Chronic exposures to environmentally relevant concentrations of glyphosate significantly impaired fecundity and fertility rates of exposed planarians (median effective concentration, EC50 = 1.6 mg glyphosate/L for fecundity and fertility rates). Our results show deleterious effects of Roundup® on regeneration, behavior and reproduction of freshwater planarians and add important ecotoxicological data towards the environmental risk assessment of glyphosate-based herbicide in freshwater ecosystems.