Effects of glyphosate on earthworms: From fears to facts.

Glyphosate is the most widely applied herbicide worldwide, contaminating water, soils, and living organisms. Earthworms are emblematic soil organisms used as indicators of soil quality, but knowledge about the impacts of glyphosate and glyphosate-based herbicides (GBH) on these key soil organisms is scattered. Here, we examine this knowledge in detail to answer four questions: (1) Which endpoint is the most sensitive when assessing the effects of glyphosate or GBH in earthworms? (2) Which is most toxic to earthworms: glyphosate or GBH? (3) Are glyphosate and GBH harmful to earthworms when used at the recommended application dose? (4) What are the interactions between glyphosate or GBH and other chemicals in earthworms? The results indicate that a weak legislation led to improper assessment of the ecotoxicity of glyphosate during the last renewal in 2017. Our findings also highlighted that negative effects can occur in earthworms at the recommended application rate, although not after only a single application or when considering only the mortality of adult individuals. However, under more realistic conditions, that is, when assessing sensitive endpoints (e.g., reproduction, growth) and using species present in the field, after several applications per year, the negative effects of glyphosate or GBH on earthworms were observed at the subindividual, individual, population, and community levels, as well as on earthworm-mediated functions. Our recommendations are as follows: (i) competent agencies should collect more information on the toxicity of these compounds to earthworms before the next renewal deadline, with emphasis on the use of the updated legislation on the topic, and (ii) scientists should increase research on the effects of these herbicides on soil invertebrate species, with emphasis on earthworms, using guideline tests and obtain data from long-term field testing. Integr Environ Assess Manag 2023;00:1-7. © 2023 SETAC.

Bringing ecology into toxicology: Life-cycle toxicity of two neonicotinoids to four different species of springtails in LUFA 2.2 natural soil.

Collembolans comprise one of the most abundant groups of soil invertebrates within the arthropods. The parthenogenetic species Folsomia candida (Willem, 1902) is the most well-studied representative, being used since the beginning of the 1960s as a model organism for assessing toxicity of chemicals in soil. In this paper we aimed at answering three questions by exposing four different species of springtails (F. candida, Folsomia fimetaria, Sinella curviseta and Heteromurus nitidus) to the neonicotinoids imidacloprid and thiacloprid: i) How representative as a model organism is F. candida for species of springtails that reproduce sexually? (ii) How suitable are other species of springtails to be used as model organisms for ecotoxicological testing? (iii) Is it possible to use the life history of these species to extrapolate the impact of neonicotinoids on the population level? Our results showed that F. candida is a good model organism, despite being the most sensitive species tested, when analysing both endpoints - survival and reproduction. The tests performed with S. curviseta and H. nitidus showed that they could be used as surrogates in ecotoxicity tests, and also to predict how their population might be affected after being exposed to chemicals. The adjustments made to the test performed with F. candida: introducing adults (20-22 days old) into the test jars and exposing them for 21 days instead of 28 days, proved to be as efficient as the standardized test guideline (OECD 232, 2009).

Toxicity in Neonicotinoids to Folsima candida and Eisenia andrei.

We compared the toxicity of the neonicotinoids imidacloprid, thiacloprid, thiamethoxam, acetamiprid, and clothianidin in terms of the survival and reproduction of 2 species of soil invertebrates, Folsomia candida and Eisenia andrei. Tests were performed using LUFA 2.2 natural soil, following standard protocols aimed at answering 2 questions: 1) Is there a difference in the toxicity between pure compound and its formulation? and 2) Is there a difference in the sensitivity of the species exposed to the same compound? For E. andrei, formulations and pure compounds had similar toxicity to both endpoints tested. For F. candida, acetamiprid and imidacloprid had different toxicities, with acetamiprid being 4 times more toxic to survival (median lethal concentration [LC50] 0.12 mg active substance [a.s.]/kg dry soil) and imidacloprid being 4 times more toxic to reproduction of the springtail (median effect concentration [EC50] 0.25 mg a.s./kg dry soil) than their commercial formulations. The most toxic compound to E. andrei was acetamiprid (LC50 0.80 and EC50 0.35-0.40 mg a.s./kg), and the most toxic to F. candida was clothianidin (LC50 0.07 and EC50 0.05 mg a.s./kg). Estimated risk ratios indicated that only one application/yr of clothianidin in the formulation Poncho® may pose a threat to the populations of springtails and earthworms. Environ Toxicol Chem 2020;39:548-555. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Assessing the toxicity of thiamethoxam, in natural LUFA 2.2 soil, through three generations of Folsomia candida.

In the field, long-term exposure is a rule rather than an exception. As a consequence, the relatively short-term standard toxicity tests may not be adequate for assessing long-term effects of pesticide exposure. This study determined the toxicity of the neonicotinoid thiamethoxam, both pure and in the formulation Actara® (25% active substance), to the springtail Folsomia candida, over three generations (P, F1 and F2). For the parental generation (P), the toxicity of pure thiamethoxam and Actara® did not differ significantly, with LC50s and EC50s of 0.32-0.35 and 0.23-0.25 mg a.s./kg dry soil, respectively. For the F1 and F2 generations, LC50s were >0.37 mg a.s./kg dry soil for both compounds. Actara was more toxic towards reproduction in the F1 generation (EC50 0.16 mg a.s./kg dry soil) than pure thiamethoxam (EC50 0.23 mg a.s./kg dry soil). For generation F2, there was no significant difference in the toxicity of the compounds towards reproduction, with EC50s of >0.37 and 0.30 mg a.s./kg dry soil for Actara® and pure thiamethoxam respectively. These results suggest a slight decrease in the toxicity of the compounds throughout the generations tested. The similarity in the toxicity of pure and formulated thiamethoxam indicates that the ingredients in the formulation Actara® do not enhance toxicity.

Comparative toxicity of imidacloprid and thiacloprid to different species of soil invertebrates.

Neonicotinoid insecticides have come under increasing scrutiny for their impact on non-target organisms, especially pollinators. The current scientific literature is mainly focused on the impact of these insecticides on pollinators and some aquatic insects, leaving a knowledge gap concerning soil invertebrates. This study aimed at filling this gap, by determining the toxicity of imidacloprid and thiacloprid to five species of soil invertebrates: earthworms (Eisenia andrei), enchytraeids (Enchytraeus crypticus), Collembola (Folsomia candida), oribatid mites (Oppia nitens) and isopods (Porcellio scaber). Tests focused on survival and reproduction or growth, after 3-5 weeks exposure in natural LUFA 2.2 standard soil. Imidacloprid was more toxic than thiacloprid for all species tested. F. candida and E. andrei were the most sensitive species, with LC50s of 0.20-0.62 and 0.77 mg/kg dry soil for imidacloprid and 2.7-3.9 and 7.1 mg/kg dry soil for thiacloprid. EC50s for effects on the reproduction of F. candida and E. andrei were 0.097-0.30 and 0.39 mg/kg dry soil for imidacloprid and 1.7-2.4 and 0.44 mg/kg dry soil for thiacloprid. The least sensitive species were O. nitens and P. scaber. Enchytraeids were a factor of 5-40 less sensitive than the taxonomically related earthworm, depending on the endpoint considered. Although not all the species showed high sensitivity to the neonicotinoids tested, these results raise awareness about the effects these insecticides can have on non-target soil invertebrates.

Multigeneration toxicity of imidacloprid and thiacloprid to Folsomia candida.

In a recent study, we showed that the springtail Folsomia candida was quite sensitive the neonicotinoid insecticides imidacloprid and thiacloprid. This study aimed at determining the toxicity of both compounds to F. candida following exposure over three generations, in natural LUFA 2.2 standard soil. In the first generation, imidacloprid was more toxic than thiacloprid, with LC50s of 0.44 and 9.0 mg/kg dry soil, respectively and EC50s of 0.29 and 1.5 mg/kg dry soil, respectively. The higher LC50/EC50 ratio suggests that thiacloprid has more effects on reproduction, while imidacloprid shows lethal toxicity to the springtails. In the multigeneration tests, using soil spiked at the start of the first generation exposures, imidacloprid had a consistent effect on survival and reproduction in all three generations, with LC50s and EC50s of 0.21-0.44 and 0.12-0.29 mg/kg dry soil, respectively, while thiacloprid-exposed animals showed clear recovery in the second and third generations (LC50 and EC50 > 3.33 mg/kg dry soil). The latter finding is in agreement with the persistence of imidacloprid and the fast degradation of thiacloprid in the test soil.