Multiple Assays on Non-Target Organisms to Determine the Risk of Acute Environmental Toxicity in Tebuconazole-Based Fungicides Widely Used in the Black Sea Coastal Area.

The widespread use of Tebuconazole-based fungicides in phytosanitary treatments on a wide range of crops, on the one hand, and the lack of official reports on the amount of fungicide residues in nearby water basins, on the other hand, may lead to uncontrolled and hazardous contamination of water sources used by the resident population, and to serious effects on the environment and public health. Our study explores the acute toxicological risk of this fungicide on various organisms, from bacteria and yeast to fish, using a battery of tests (standardized Toxkit microbiotests and acute semi-static tests). By investigating the interaction between Tebuconazole and bacteria and yeast organisms, we observed that Gram-negative bacteria displayed a strong tolerance for Tebuconazole, while Gram-positive bacteria and yeasts proved to be very sensitive. The fish experiment was conducted on Chelon auratus juveniles exposed to five concentrations of the fungicide Tebustar EW (Tebuconazole, 250 g/L as active substance). After 96 h of exposure, the LC50 for C. auratus was 1.13 mg/L. In the case of the Toxkit microbiotests' application, the following results were recorded: Spirodela polyrhiza EC50 = 2.204 mg/L (after 72 h exposure), Thamnocephalus platyurus EC50 = 0.115 mg/L (after 24 h), and Daphnia magna EC50 = 2.37 mg/L (after 24-48 h). With the exception of bacteria and yeast, the same response pattern was observed for all non-target species tested; the response range expressed by concentrations causing growth inhibition or mortality was small, ranging between very close values that are quite low, thereby demonstrating the high toxicity of Tebuconazole-based fungicides to the environment.

A Multi-Community Approach in Biodiversity Assessment of a Peat Bog in the Southern Carpathians (Romania) and Implications for Conservation.

Although natural peatlands have been recognized as an important type of wetlands because they support high biodiversity and provide important ecosystem services, the value of peatlands both in biodiversity research and conservation is still largely underestimated. Our study characterizes the biodiversity and conservation value of Pesteana peat bog, an upland mesotrophic peat bog, located in the Southern Carpathians, Romania. More specifically, we: (1) characterized the invertebrate (i.e., top soil, surface litter, and plant-dwelling) and plant communities along a humidity gradient in Pesteana peat bog and nearby habitats (i.e., treeline, ecotone, lowland and highland meadow, and forest), (2) assessed the main environmental factors driving the invertebrate community diversity and composition, and (3) determined the relationship between invertebrate community diversity and vegetation, focusing on the top soil invertebrates. Our study revealed a high diversity of invertebrates spanning over 43 taxonomic groups and a high number of plant indicator species, emphasizing the role of natural peatlands in preserving diverse communities in a small area. The results showed that the composition of top soil invertebrate community was determined by depth of organic layer, vegetation cover, and soil compaction. We found that the diversity of top soil invertebrate community was strongly influenced by habitat type and soil attributes and weakly by vegetation. Overall, the invertebrate and plant communities showed different responses to habitat conditions along the humidity gradient. This highlights the importance of using a multi-community approach to support the design of effective conservation and management actions beneficial for a wide range of taxa.

Ecotoxicological Risk Assessment of Actellic 50 EC Insecticide on Non-Target Organisms in Parallel with the Application of Standardized Tests.

This paper contributes to the ecotoxicological risk assessment of the Actellic 50 EC insecticide (with 50% pirimiphos-methyl as the active substance) tested on non-target organisms. The insecticide concentrations tested were the same for all organisms (0.1, 0.01, and 0.001 mg L-1 of Actellic 50 EC), with an exposure of 3-5-21 days for plants and 4-5-14 days for animals. The non-target organisms tested were both plants (wheat and two ferns) and animals (the Prussian carp and marsh frog tadpoles). The tested insecticide significantly inhibited the growth of roots in wheat, a result that was also confirmed by a microbiotest application (62% root growth inhibition in sorghum and 100% germination inhibition in white mustard and garden cress). In ferns, even for the lowest concentration, the percentage of germinated spores was inhibited by 40% for Asplenium scolopendrium. The recorded toxicological effects of Actellic 50 EC upon the Prussian carp included a decrease in the respiratory rate and oxygen consumption, an increase in the number of erythrocytes and leukocytes, and an increase in blood glucose levels. The highest concentration (0.1 mg L-1 of Actellic 50 EC) caused a 50% decrease in the survival rate of marsh frog tadpoles after 5 days of exposure, negatively affecting body volume and length. Given the high degree of toxicity of the insecticide Actellic 50 EC, we recommend continuing investigations on non-target species, including both plants and animals, as the sub-chronic effects are quite little known in the scientific literature.

Ecotoxicological Studies on the Action of Actara 25 WG Insecticide on Prussian Carp (Carassius gibelio) and Marsh Frog (Pelophylax ridibundus).

The toxic action of the Actara 25 WG insecticide (it contains 25% thiamethoxam as an active substance) in non-lethal doses was studied in two species of aquatic organisms-the Prussian carp (Carassius gibelio) and the marsh frog (Pelophylax ridibundus)-at two thermal levels, 6-8 °C (low temperature) and 18-20 °C (room temperature), respectively. In the Prussian carp, we recorded decreases in oxygen consumption and stimulation of the respiratory rhythm, changes that were more pronounced in the case of intoxicated fish and when the species were kept at room temperature. The histopathology of the lung in the frog illustrated the thickening of the conjunctival septum, an increase in the number of mucous cells, and an increase in the ratio between the diameter of the nucleus and the diameter of the pneumocyte. All of these changes were more pronounced in the animals kept at higher temperature. Our study looks at the extent to which temperature changes can influence the ability of poikilothermic organisms to withstand the presence of toxic substances in the environment as a result of the impact of the use of insecticides in agriculture. The two tested organisms are a common presence for the study area, which was affected in the last decade by climate change.

Bioaccumulation of Copper and Zinc and the Effects on Antioxidant Enzyme Activities in the Liver of Acipenser stellatus (Pallas, 1771).

Although water pollution by metals in the Danube River is considered high, little is known about its impact on sturgeons. In this regard, the aim of this study was to investigate the bioaccumulation of copper and zinc as well as their effects on antioxidant enzymes activities in the liver of Acipenser stellatus. The fish were exposed for 7 and 14 days, to two concentrations of copper and zinc (10% and 25% of LC50 96 h), previously determined as 0.54 mg/L Cu2+ and, 34.22 mg/L Zn2+ respectively. The enzymatic responses of A. stellatus varied greatly depending on metal type, concentration and time. Significant bioaccumulation of the two metals was recorded. Even though the water hardness used in the experiment is known to offer a clear protection against metal contamination, stellate sturgeon remains a sensitive species in the face of metal toxicity.

A practical and user-friendly toxicity classification system with microbiotests for natural waters and wastewaters.

Various types of toxicity classification systems have been elaborated by scientists in different countries, with the aim of attributing a hazard score to polluted environments or toxic wastewaters or of ranking them in accordance with increasing levels of toxicity. All these systems are based on batteries of standard acute toxicity tests (several of them including chronic assays as well) and are therefore dependent on the culturing and maintenance of live stocks of test organisms. Most systems require performance of the bioassays on dilution series of the original samples, for subsequent calculation of L(E)C50 or threshold toxicity values. Given the complexity and costs of these toxicity measurements, they can only be applied in well-equipped and highly specialized laboratories, and none of the classification methods so far has found general acceptance at the international level. The development of microbiotests that are independent of continuous culturing of live organisms has stimulated international collaboration. Coordinated at Ghent University, Belgium, collaboration by research groups from 10 countries in central and eastern Europe resulted in an alternative toxicity classification system that was easier to apply and substantially more cost effective than any of the earlier methods. This new system was developed and applied in the framework of a cooperation agreement between the Flemish community in Belgium and central and eastern Europe. The toxicity classification system is based on a battery of (culture-independent) microbiotests and is particularly suited for routine monitoring. It indeed only requires testing on undiluted samples of natural waters or wastewaters discharged into the aquatic environment, except for wastewaters that demonstrate more than 50% effect. The scoring system ranks the waters or wastewaters in 5 classes of increasing hazard/toxicity, with calculation of a weight factor for the concerned hazard/toxicity class. The new classification system was applied during 2000 by the participating laboratories on samples of river water, groundwaters, drinking waters, mine waters, sediment pore waters, industrial effluents, soil leachates, and waste dump leachates and was found to be easy to apply and reliable.