Arbuscular Mycorrhizal Fungi and the Need for a Meaningful Regulatory Plant Protection Product Testing Strategy.

Arbuscular mycorrhizal fungi (AMF) perform key soil ecosystem services and, because of their symbiotic relationship with plant roots, may be exposed to the plant protection products (PPPs) applied to soils and crops. In 2017, the European Food Safety Authority (EFSA) released a scientific opinion addressing the state of the science on risk assessment of PPPs for in-soil organisms, recommending the inclusion of AMF ecotoxicological testing in the PPP regulatory process. However, it is not clear how this can be implemented in a tiered, robust, and ecologically relevant manner. Through a critical review of current literature, we examine the recommendations made within the EFSA report and the methodologies available to integrate AMF into the PPP risk assessment and provide perspective and commentary on their agronomic and ecological relevance. We conclude that considerable research questions remain to be addressed prior to the inclusion of AMF into the in-soil organism risk assessment, many of which stem from the unique challenges associated with including an obligate symbiont within the PPP risk assessment. Finally, we highlight critical knowledge gaps and the further research required to enable development of relevant, reliable, and robust scientific tests alongside pragmatic and scientifically sound guidance to ensure that any future risk-assessment paradigm is adequately protective of the ecosystem services it aims to preserve. Environ Toxicol Chem 2022;41:1808-1823. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Ecotoxicological test to assess effects of herbicides on spore germination of Rhizophagus clarus and Gigaspora albida.

Given the essential role of arbuscular mycorrhizal fungi (AMF) in soil systems and agriculture, their use as biological indicators has risen in all fields of microbiology research. However, AMF sensitivity to chemical pesticides is poorly understood in field conditions, and not explored in ecotoxicology protocols. Hence, the goal of this study was to evaluate the effects of different concentrations of glyphosate (Roundup®) and diuron+paraquat (Gramocil®) on the germination of spores of Gigaspora albida and Rhizophagus clarus in a tropical artificial soil. This study was conducted in 2019 at the Soil Ecology and Ecotoxicology Laboratory of the Universidade do Estado de Santa Catarina. The nominal concentrations of glyphosate were 0, 10, 50, 100, 250, 500, 750 and 1000 mg a.i. kg-1. For diuron+paraquat, the concentrations tested were 0, 10 + 20, 50 + 100, 100 + 200, 250 + 500, 500 + 1000, 750 + 1500 and 1000 + 2000 mg a.i. kg-1. Glyphosate did not alter germination of G. albida, but germination inhibition of R. clarus spores was of 30.8% at 1000 mg kg-1. Diuron+paraquat inhibited by 8.0% germination of G. albida, but only at the highest concentration tested. On the other hand, effects on R. clarus were detected at 50 + 100 mg kg-1 concentration and above, and inhibition was as high as 57.7% at the highest concentration evaluated. These results suggest distinct response mechanisms of Rhizophagus and Gigaspora when exposed to herbicides, with the former being more sensitive than the later.

Are there any risks of the disposal of pesticide effluents in soils? Biobed system meets ecotoxicology ensuring safety to soil fauna.

The biobed is a purification system, which reduces soil pollution for receiving pesticide residues from handling and washing machinery in agricultural areas. The aims of this study were (1) to assess ecotoxicity effects over time to soil fauna, posed by Lorsban® 480 BR (Chlorpyrifos) and Dithane® NT (Mancozeb) residues when disposed of in a biobed system compared with two subtropical soils, and (2) to assess ecotoxicity effects over time to soil fauna simulating an accidental spillage with Lorsban® 480 BR at the biobed. A semi-field experiment was conducted for 420 days in southern Brazil, testing continuous disposal of washing pulverization tanks in biobeds, Typic Haploperox or Typic Hapludults. In addition, different biobeds received a single dose (1 L) of Lorsban® 480 BR to simulate an accidental spillage. Chronic ecotoxicity tests were performed using Folsomia candida, Eisenia andrei, and Enchytraeus crypticus in different sampling times for both experiments. F. candida was the most sensitive species. The biobed system was able to eliminate effects from residues of both pesticides over time in all species, which did not happen in both natural soils. In accidental spillage simulation, even 420 days after contamination, F. candida did not show reproduction. The biobeds can be a feasible alternative for the disposal and treatment residues of pesticides, also for handling and washing pesticides activities. The system was efficient in promoting degradation and reducing ecotoxicity effects posed by Lorsban® 480 BR and Dithane® NT for soil fauna. It is a safe alternative to avoid soil contamination.

Boric acid as reference substance for ecotoxicity tests in tropical artificial soil.

Reference substances are recommended to evaluate the quality of laboratory test species and the reliability of ecotoxicity data. Boric acid (BA) has been recommended as reference substance in some standardized tests in OECD soil, but no data are available for Tropical Artificial Soil (TAS). For this purpose, avoidance tests with Eisenia andrei, lethality tests with E. andrei and Folsomia candida, and reproduction tests with E. andrei, Enchytraeus crypticus and F. candida were carried out in TAS (5% organic matter), following ISO guidelines, and compared between two laboratories. Collembolans were more sensitive than earthworms in lethality tests (LC50 = 342 and > 1000 mg kg-1, respectively). For both laboratories, the EC50 values were similar for reproduction of oligochaeta species (165 mg kg-1 for E. crypticus; 242 and 281 mg kg-1 for E. andrei), but significantly different for reproduction of F. candida (96 and 198 mg kg-1). Present results suggest that boric acid could replace the current pesticides recommended by ISO guidelines as reference substances on reproduction tests with soil invertebrates in TAS. Concerning avoidance tests, additional investigations should be performed with other substances that cause no neurotoxic effects on soil organisms.