Toxicity of fungicides to terrestrial non-target fauna - Formulated products versus active ingredients (azoxystrobin, cyproconazole, prothioconazole, tebuconazole) - A case study with Enchytraeus crypticus (Oligochaeta).

Despite the high usage of pesticides in current agricultural practices, its effects to humans and to the environment (non-target species) are a continuous concern. Soil dwelling organisms are among the first in line of exposure to pesticides, however their risks are often based on the pure active ingredient (a.i.) and not on the commercial formulated products (FPs) actually applied in the fields. In the present study, we investigated the effects of two fungicide FPs versus its a.i. (s): Amistar® XTRA and the respective a.i. (s) azoxystrobin and cyproconazole, and Prosaro® 250 EC and the respective a.i. (s) prothioconazole and tebuconazole, to the non-target soil oligochaete Enchytraeus crypticus. The standard Enchytraeid Reproduction Test was used to assess effects on survival and reproduction. Results showed that Amistar was more toxic than Prosaro, particularly for reproduction (EC50 = 161 mg Amistar/kg soil, EC50 = 350 mg Prosaro/kg soil). For both FPs, reproductive effects were mainly related to one of its a.i. (s) (azoxystrobin [EC50 = 37 mg azosxystrobin/kg soil] for Amistar, and tebuconazole [EC50 = 41 mg tebuconazole/kg soil] for Prosaro), while lethal effects were not predicted by the toxicity of its a.i. (s) (particularly in the case of Prosaro, which was more toxic than its a.i. (s)). These findings highlight the need to further explore the toxicity data of the FPs compared to the a.i. (s), aiming to predict a more realistic environmental hazard of pesticides.

Multi-biomarker approach and IBR index to evaluate the effects of different contaminants on the ecotoxicological status of Apis mellifera.

The honeybee, Apis mellifera L. (Hymenoptera: Apidae), a keystone pollinator of wild plant species and agricultural crops, is disappearing globally due to parasites and diseases, habitat loss, genetic constraints, beekeeper management issues and to the widespread use of pesticides. Besides insecticides, widely studied in this species, honeybees are also exposed to herbicides and fungicides and heavy metals whose lethal and sublethal effects need to be investigated. In this context, our study aimed to evaluate the effects of fungicides and of heavy metals on honeybees and to develop and apply a multi-biomarker approach that include an Integrated Biological Index (IBRv2) to assess the toxicological status of this species. Biomarkers of neurotoxicity (AChE and CaE), metabolic alteration (ALP, and GST) and immune system (LYS, granulocytes) were measured, following honeybees' exposure to cadmium or to a crop fungicide, using the genotoxic compound EMS as positive control. A biomarker of genotoxicity (NA assay) was developed and applied for the first time in honeybees. At the doses tested, all the contaminants showed sublethal toxicity to the bees, highlighting in particular genotoxic effects. The data collected were analyzed by an IBRv2 index, which integrated the seven biomarkers used in this study. IBRv2 index increased with increasing cadmium or fungicide concentrations. The IBRv2 represents a simple tool for a general description of honeybees ecotoxicological health status. Results highlight the need for more in-depth investigations on the effects of fungicides on non-target organisms, such as honeybees, using sensitive methods for the determination of sublethal effects. This study contributes to the development of a multi-biomarker approach to be used for a more accurate ecotoxicological environmental monitoring of these animals.

Apis mellifera ligustica, Spinola 1806 as bioindicator for detecting environmental contamination: a preliminary study of heavy metal pollution in Trieste, Italy.

Honeybees have become important tools for the ecotoxicological assessment of soil, water and air metal contamination due to their extraordinary capacity to bioaccumulate toxic metals from the environment. The level of heavy metal pollution in the Trieste city was monitored using foraging bees of Apis mellifera ligustica from hives owned by beekeepers in two sites strategically located in the suburban industrial area and urban ones chosen as control. The metal concentration in foraging bees was determined by inductively coupled plasma-mass spectrometry. The chemical analysis has identified and quantified 11 trace elements accumulated in two different rank orders: Zn> Cu > Sr > Bi > Ni > Cr > Pb = Co > V > Cd > As in foraging bees from the suburban site and Zn > Cu > Sr > Cr > Ni > Bi > Co = V > Pb > As > Cd in bees from urban site. Data revealed concentrations of Cr and Cu significantly higher and concentration of Cd significantly lower in bees from urban sites. The spatial difference and magnitude order in heavy metal accumulation along the urban-suburban gradient are mainly related to the different anthropogenic activity within sampled sites and represent a risk for the human health of people living in the city. We discussed and compared results with the range of values reported in literature.