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Abstract: Abstract The hyporheic zone, i.e. the water-saturated sediment beneath and alongside the riverbed, is exposed to multiple stressors. Agricultural-watershed rivers are frequently exposed to two concomitant stressors: clogging and copper contamination. However, one stressor exposure can increase sensitivity to a second stressor. The aim of this study was to experimentally test the cumulative effects of these two stressors on copper distribution and structural and functional microbial communities responses in the hyporheic zone. A slow filtration column experiment was conducted to compare the effects of 3 treatments of increasing complexity: ‘Reference’, ‘Copper-contaminated’ (dissolved copper added at 191 µg L−1), and ‘Clogging+Copper’ (dissolved copper + addition of 2 cm of fine sediment). Microbial community structure and activities were studied at 4 column sediment depths. The results showed that clogging did not modify the distribution of copper, which remained fixed in the first few centimetres. In the first few centimetres, clogging had a stimulating effect on microbial activities whereas copper had limited effects mainly on leucine aminopeptidase activity and microbial community tolerance to copper. The subsurface zone thus hosts significant different microbial communities from the communities in the deeper zones that were protected from surface stressors. This experiment confirms the valuable filtering role played by the hyporheic zone and shows that microbial responses are strongly correlated to microhabitat-scale physicochemical conditions in sediment. PubDate: 2023-03-17
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Abstract: Abstract Imazethapyr, a post-emergent herbicide used in worldwide soybean and corn crops, induces genetic and biochemical alterations in aquatic vertebrates. This study examined the relationship between biomarkers at different organization levels and imazethapyr real-life route exposure in Boana pulchella adults. Frogs were exposed to imazethapyr-based formulation Pivot® H (10.59%) at concentrations representing possible acute routes: field runoff (S1:10 mg.L−1), exposure after direct foliar application (S2:100 mg.L−1) and during direct foliar application (S3:1000 mg.L−1). Post-exposure, endpoints levels were evaluated: organism alterations, biochemical activities and cytogenetic assays. Forty-eight hours post-exposure, antioxidant enzymes decrease, micronuclei induction and DNA damage were observed in all scenarios, while cholinesterase activity increase and body condition reduction were observed in frog-exposed to S3. Ninety-six hours post-exposure, frogs showed glutathione-S-transferase inhibition in S1, micronuclei induction in S2 and S3, and DNA-damage increase in S3. Herbicides routes of exposures in real-life could indicate that authorized applications have a risk to amphibian populations. PubDate: 2023-03-16
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Abstract: Abstract Carabids (Coleoptera: Carabidae) are abundant predators in ecosystems and serve as pest biocontrol in agroecosystems and forestry. Here we test the impact of thiamethoxam, among the most used neonicotinoids on the consumption rate, locomotion, metabolomics, and oxidative stress level measuring superoxide dismutase (SOD) activity in a predatory carabid, Abax parallelus (Duftschmid, 1812), after acute exposure in the laboratory trials, to get additional data that might link the use of pesticides and predation efficiency. Beetles were exposed to increasing concentrations of thiamethoxam by dipping method, and left to feed overnight prior to the assays. The results showed that individuals treated with higher concentrations of thiamethoxam (20 and 40 mg/L) consumed significantly less food per body weight and had a higher share of intoxicated and moribund individuals. The mass of consumed food per beetle body weight and observed locomotion did not differ significantly between control and groups treated with lower concentrations of thiamethoxam. There are significant differences in concentrations of some metabolites between treated and control individuals, primary in succinate and d-glucose, indicating a disruption in energy production. On the other hand, there is no statistically significant differences in SOD activity among the groups. To conclude, acute exposure to thiamethoxam can result in negative sub-lethal effects in predatory activity and energy budget, while the effects of long-term exposure to lower doses require further research, as well as field assessment on the predation efficiency after pesticide application. PubDate: 2023-03-11
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Abstract: Abstract Microplastic (MP) is yet another form of chronic anthropogenic contribution to the environment. MPs are plastic particles (<5 mm) that have been widely found in the most diverse natural environments, but their real impacts on ecosystems are still under investigation. Here, we studied the toxicity of naturally aged secondary polypropylene (PP) MPs after constant exposure to ultraviolet radiation (26 µm) to the third instar larvae of Chironomus sancticaroli, a dipteran species. The concentrations tested were 13.5; 67.5; and 135 items g−1 of dry sediment. C. sancticaroli organisms were investigated for fragment ingestion, mortality and changes in enzymatic biomarkers after 144 h of exposure. The organisms were able to ingest MPs from the first 48 h, and the amount of items internalized was dose-dependent and time-dependent. Overall, the results show that mortality was low, being significant at the lowest and highest concentrations (13.5 and 135 items g−1). Regarding changes in biochemical markers, after 144 h MDA and CAT activities were both significantly altered (increased and reduced, respectively), while SOD and GST levels were unchanged. In the present study, naturally aged polypropylene MPs induced biochemical toxicity in C. sancticaroli larvae, with toxicity being higher according to exposure time and particle concentration. PubDate: 2023-03-11
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Abstract: Abstract Ecological risk assessment of chemicals focuses on the response of different taxa in isolation not taking ecological and evolutionary interplay in communities into account. Its consideration would, however, allow for an improved assessment by testing for implications within and across trophic levels and changes in the phenotypic and genotypic diversity within populations. We present a simple experimental system that can be used to evaluate the ecological and evolutionary responses to chemical exposure at microbial community levels. We exposed a microbial model system of the ciliate Tetrahymena thermophila (predator) and the bacterium Pseudomonas fluorescens (prey) to iron released from Magnetic Particles (MP-Fedis), which are Phosphorus (P) adsorbents used in lake restoration. Our results show that while the responses of predator single population size differed across concentrations of MP-Fedis and the responses of prey from communities differed also across concentration of MP-Fedis, the community responses (species ratio) were similar for the different MP-Fedis concentrations. Looking further at an evolutionary change in the bacterial preys’ defence, we found that MP-Fedis drove different patterns and dynamics of defence evolution. Overall, our study shows how similar community dynamics mask changes at evolutionary levels that would be overlooked in the design of current risk assessment protocols where evolutionary approaches are not considered. PubDate: 2023-03-04
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Abstract: Abstract Soil sorption properties can influence the bioavailability of substances and consequently the toxicity for soil organisms. Current standardised laboratory testing for the exposure assessment of pesticides to soil organisms uses OECD artificial soil that does not reflect the high variation in chemical-physical soil properties found in natural agroecosystems. According to guideline OECD 222, earthworm reproduction tests with Eisenia fetida and the pesticide carbendazim were performed in four natural soils and OECD artificial soil. By using pF 1.6, which ensures a uniformity in actual soil water availability, the control reproduction performance of E. fetida in all natural soils was at the same level as OECD artificial soil. In a principle component analysis, the variation in toxicity between the tested soils was attributable to a combination of two soil properties, namely total organic carbon content (TOC) and pH. The largest difference of 4.9-fold was found between the typical agricultural Luvisol with 1.03% TOC and pH 6.2 (EC10: 0.17 (0.12–0.21) mg a.i. kg−1 sdw, EC50: 0.36 (0.31–0.40) mg a.i. kg−1 sdw) and OECD artificial soil with 4.11% TOC and pH 5.6 (EC10: 0.84 (0.72–0.92) mg a.i. kg−1 sdw, EC50: 1.07 (0.99–1.15) mg a.i. kg−1 sdw). The use of typical agricultural soils in standardised laboratory earthworm testing was successfully established with using the measure pF for soil moisture adjustment. It provides a more application-oriented approach and could serve as a new tool to refine the environmental risk assessment at lower tier testing or in an intermediate tier based approach. PubDate: 2023-03-01
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Abstract: Abstract Mexico City is one of the most polluted cities in the world, and one in which air contamination is considered a public health threat. Numerous studies have related high concentrations of particulate matter and ozone to several respiratory and cardiovascular diseases and a higher human mortality risk. However, almost all of those studies have focused on human health outcomes, and the effects of anthropogenic air pollution on wildlife species is still poorly understood. In this study, we investigated the impacts of air pollution in the Mexico City Metropolitan Area (MCMA) on house sparrows (Passer domesticus). We assessed two physiological responses commonly used as biomarkers: stress response (the corticosterone concentration in feathers), and constitutive innate immune response (the concentration of both natural antibodies and lytic complement proteins), which are non-invasive techniques. We found a negative relationship between the ozone concentration and the natural antibodies response (p = 0.003). However, no relationship was found between the ozone concentration and the stress response or the complement system activity (p > 0.05). These results suggest that ozone concentrations in air pollution within MCMA may constrain the natural antibody response in the immune system of house sparrows. Our study shows, for the first time, the potential impact of ozone pollution on a wild species in the MCMA presenting the Nabs activity and the house sparrow as suitable indicators to assess the effect of air contamination on the songbirds. PubDate: 2023-02-21
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Abstract: Abstract Port areas are socio-ecosystems impacted by chronic mixture pollution. Some marine species benefit from living there and may be studied to define the ecological state of such environments. In this study, the risks of chronic chemical contamination and its consequences on three marine molluscs were evaluated in North Corsica (France) port areas. Mediterranean mussel Mytilus galloprovincialis, tubular sea cucumber Holothuria tubulosa and Mediterranean limpet Patella sp. were sampled in three port areas and a reference location. A set of biomarkers was analysed to evaluate oxidative stress, detoxification, energetic metabolism, neurotoxicity, immunity and bioaccumulation (metallic trace elements and organic pollutants). The objectives were to assess pollution-induced effects in organisms, to determine the best bioindicator species for the selected locations and to validate a “pool” sampling technique (when the analysis is done on a single pool of samples and not on individual samples). The results validate the sampling techniques as “pool” for management purposes. St-Florent was demonstrated as the most contaminated location. All the other locations present a low contamination, below the recommended threshold values (for metallic trace elements and organic pollutants). Finally, the limpet appears to be the best bioindicator for the selected locations. Mussel and sea cucumber are inappropriate due to their absence in this oligotrophic region and the lack of responses observed, respectively. PubDate: 2023-02-17
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Abstract: Abstract Apis mellifera is an important bee pollinating native and crop plants but its recent population decline has been linked to the use of pesticides, including fungicides that have been commonly classified as safe for bees. However, many pesticides, in addition to direct mortality cause sublethal effects, including damage to target selective honey bee organs. The midgut is the organ responsible for the digestion and absorption of nutrients and the detoxification of ingested substances, such as pesticides. This study evaluated the histopathological and cytotoxic changes in the midgut of A. mellifera workers caused by the pesticide azoxystrobin. The limit-test was performed, and a 100 µg a.i./bee dose was administered orally and midgut analyzed with light and transmission electron microscopies after 24 h and 48 h of pesticide exposure. The midgut of the control bees has a single layer of digestive cells, with spherical nuclei, nests of regenerative cells, and the lumen coated with the peritrophic matrix. The bees fed on azoxystrobin showed morphological changes, including intense cytoplasm vacuolization and cell fragments released into the gut lumen. The protein detection test showed greater staining intensity in the nests of regenerative cells after 24 h of exposure to azoxystrobin. The occurrence of damage to the midgut in A. mellifera exposed to azoxystrobin indicates that although this fungicide has been classified as low toxicity for bees, it has sublethal effects in the midgut, and effects in other organs should be investigated. PubDate: 2023-02-06
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Abstract: Abstract In recent years, there has been an increase in research to understand the consequences of microplastic contamination. A subset of this research assesses the interaction of microplastics with metals and the subsequent effects of the resulting microplastic-metal complexes in freshwater environments. While our understanding of how microplastics behave in freshwater remains largely unknown, our knowledge of metal behavior in those same environments is well-established. The behavior (partitioning, speciation, bioavailability) of metals is highly dependent on environmental characteristics, including water quality variables such as hardness, pH, and dissolved organic matter. This study reveals that despite our understanding of metal behavior, there is little consideration for these influential factors in the current body of microplastic-metal research. Multiple instances highlighted throughout this study show that even when similar plastic, metal, and biota are utilized, there are conflicting observations as to whether the mixture is toxic; we stress that without adequate reporting of environmental conditions, these contradictions are likely to persist without explanation. Through justification of water quality characteristics known to influence metal behavior, this study proposes a framework of reporting requirements for all future microplastic-metal research. PubDate: 2023-02-06
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Abstract: Abstract Chironomus sancticaroli is a tropical species, easy to grow and to maintain in laboratory cultures. It has a fast reproduction cycle, under adequate conditions, around 30 days, allowing it to have many generations per year, an important criterion for selecting a test organism in ecotoxicology. Its life stages include: eggs, four larval instars (one planktonic and three benthic), pupa and adult (midges) This study aimed to: (1) review the methods for C. sancticaroli cultivation and its use in ecotoxicological tests, (2) establish a laboratory culture of C. sancticaroli, presenting the difficulties and discussing the ways to overcome them. Early 4th instar larvae was the most used in acute studies, while the 1st instar larvae (early 1st instar) was the most used in chronic studies; 96 h and 28 days were the most frequent durations in acute and chronic studies, respectively. The most common endpoints evaluated were organisms’ survival and development, and most of the ecotoxicological studies using C. sancticaroli were performed in laboratory. Most of the tested contaminants were pesticides and these had the most adverse effects on organisms. Most mesocosms with environmental contaminated samples did not show adverse effects on C. sancticaroli. Chronic and field studies as well as those testing the effects of the mixture contaminants on C. sancticaroli were still deficient. Keeping the laboratory environment and equipment effectively sanitized was important as well as maintaining stabilized conditions of temperature, photoperiod, physical, chemical and biological water quality in cultures. PubDate: 2023-01-28
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Abstract: Abstract Environmental contaminant-induced aquatic pollution threatens the aquatic biota and causes oxidative stress in the aquatic organisms. Mahananda River, the lifeline of Malda district, West Bengal, India is presently experiencing environmental crisis due to increasing level of contaminants which deteriorate the water quality along with its biota. Puntius sarana is an economically important edible fish species in the River Mahananda that seems to be declining, owing to environmental pollution. Therefore, the present study aimed to investigate the health status of Puntius sarana by evaluating different oxidative stress biomarkers. The sampling sites were selected according to population density and possible anthropogenic activities around the River Mahananda. Significantly higher concentration of Zn2+ (p < 0.01), Cu2+ (p < 0.05) in different sampling sites has been observed, compared to the control site. Results of the water quality index and water pollution index confirmed the pollution status of different sampling sites. Significant decreases were observed in superoxide dismutase and catalase activity, compared to the control. The decreased values of reduced and oxidized glutathione ratio also indicate the increased oxidative stress of Puntius sarana. The principal component analysis showed that total hardness, calcium hardness, nitrate, manganese, zinc, copper, iron and arsenic were the main descriptors of the aquatic pollution. Therefore, findings of the present study demonstrate the ecotoxicological impacts of various anthropogenic activities which necessitates the need for continuous monitoring of the fresh water resources and its ecosystem from further contamination. PubDate: 2023-01-28
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Abstract: Abstract Profenofos (organophosphate) is among the major toxicant polluting freshwater bodies, exerting a significant effect on fish health. The LC50 value of Profenofos (PRO) was resolved in Grass carp (Ctenopharyngodon idella) with average body weight (55.82 ± 5.42 g) and determined the 96 h LC50 value as 7.2 µg/L for the assay. Twenty-one-day exposures to 1.8 µg/ L and 3.6 µg/ L doses were conducted to evaluate the sub-lethal effects, and various toxicological endpoints were assessed on the 1st, 7th, 15th and 21st days of exposure. Acute toxic stress was observed with fish displaying behavioral toxicity. The most hematological change was extreme microcytic hypochromic anemia. Leucocyte count increased in experimented fish. Moderate neutrophilia, monocytosis and lymphocytosis were observed. Serum total protein, albumin, and globulin concentrations were significantly diminished. Overall, increments over control were recognized in serum urea, creatinine and acid phosphatase. However, serum glucose, total lipid, cholesterol, serum ALT and AST activity showed a significant decrease in fish exposed to both concentrations of PRO. Serum IgM concentrations insignificantly changed in treated fish except for on the 21st day of exposure to 3.6 µg/ L of PRO, while serum lysozyme significantly decreased. Furthermore, total protein, lipid and glycogen concentrations in muscles and the liver exhibited a decreasing trend at all concentrations. Moreover, histopathological alterations in the liver, kidney, and muscles occurred exclusively after treatment. From the obtained results, it is assumed that profenofos induced general toxic impacts under field conditions and might disturb ecologically relevant processes. PubDate: 2023-01-28
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Abstract: Abstract Fungicide usage has increased globally in response to the rise in fungal pathogens, especially in the agricultural sector. However, research examining the toxicity of fungicides is still limited for many aquatic species. In this study, we examined the acute toxicity of two widely used fungicides, chlorothalonil and pyraclostrobin, on six North American larval amphibian species across multiple families using 96-h LC50 tests. We found that pyraclostrobin was approximately 3.5x more toxic than chlorothalonil; estimated LC50 values ranged from 5–18 µg/L for pyraclostrobin and 15–50 µg/L for chlorothalonil. Comparing across amphibian groups, we found that salamanders were 3x more sensitive to pyraclostrobin than anuran species and equally as sensitive to chlorothalonil. Notably, our estimated LC50 values within the range of the expected environmental concentration for these fungicides suggesting environmental exposures could lead to direct mortality in these species. Given the widespread and increasing usage of fungicides, additional work should be conducted to assess the general risk posed by these chemicals to amphibian and their associated aquatic habitats. PubDate: 2023-01-24
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Abstract: Abstract Aquatic ecosystems have been suffering deleterious effects due to the development of different economic activities. Metal(loid)s are one of the most persistent chemicals in environmental reservoirs, and may produce adverse effects on different organisms. Since fishes have been largely used in studies of metal(loid)s exposure, tilapia and largemouth bass were collected in three ecosystems from the Yaqui River Basin to measure the concentrations of metal(loid)s (chromium (Cr), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), zinc (Zn) arsenic (As), mercury (Hg), and selenium (Se)) and some biomarkers (somatic indices, metallothionein expression and histopathological analysis) in tissues of both species. Metal(loid) concentrations varied seasonally among ecosystems in tissues of both species. The elements varied seasonally and spatially in tissues of both species, with a general distribution of liver > gills > gonads. Also, biomarkers showed variations indicative that the fish species were exposed to different environmental stressor conditions. The highest values of some biomarkers were in largemouth bass, possibly due to differences in their biological characteristics, mainly feeding habits. The multivariate analysis showed positive associations between metal(loid)s and biomarkers, which are usually associated to the use of these elements in metabolic and/or regulatory physiological processes. Both fish species presented histological damage at different levels, from SI types (changes that are reversible for organ structure) to SII types (changes that are more severe but may be repairable). Taken together, the results from this study suggest that the Yaqui River Basin is moderately impacted by metals and metalloids. PubDate: 2023-01-23
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Abstract: Abstract Current international legislation regarding agrochemicals requires thorough toxicological testing mainly of the active ingredients. In a 96-h acute toxicity test we exposed Rana dalmatina and Bufo bufo tadpoles to either one of three concentrations of glyphosate, three concentrations of the surfactant (POEA), three concentrations of the two components together, or to non-contaminated water (control), and subsequently assessed mortality and body mass. To investigate whether simultaneous exposure to another stress factor influences effects of the contaminants, we performed tests both in the presence or absence of predator chemical cues. We found that the surfactant had significant harmful effects on tadpoles; survival was lowered by the highest concentration of the surfactant in case of R. dalmatina, while in B. bufo tadpoles it reduced survival already at medium concentrations. Body mass was significantly influenced by medium and high surfactant concentrations in both species. The presence of glyphosate did not have a significant effect by itself, but it slightly increased mortality in tadpoles exposed to medium concentrations of the surfactant in both species. The presence of chemical cues did not have an effect on the examined variables. Our study confirms that the toxicity of glyphosate-based herbicides is mainly due to the examined surfactant. Nonetheless, we found that glyphosate can enhance the harmful effect of the surfactant. These results stress that during the authorization process of new pesticide formulations, not only the active ingredients would need to be examined but the excipients should also be taken into account in an obligatory and systematic manner. PubDate: 2023-01-21
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Abstract: Abstract Aspirin (acetylsalicylic acid, ASA), a widely used non-steroidal anti-inflammatory drug, was frequently detected in aquatic environments around the world. However, information on the potential toxic effects of aspirin on non-target aquatic invertebrates is limited. In the present study, we investigated the effects of ASA on the transcriptional expressions of antioxidant genes (Nrf2, Keap1, HO-1, GCLC, GPx, TRX, TrxR and Prx1) and DNA methylation genes (DNMT1, DNMT3 and TET2) in Daphnia magna (D. magna)for 24, 48 and 96 h and the changes of antioxidant enzymatic activity and GSH, MDA content for 48 h. The effects of ASA on the life traits of D. magna were also addressed via a 21-days chronic toxicity test. Results showed that the expressions of Nrf2 and its target genes (HO-1, GPx and TrxR, GCLC, TRX and Prx1) were induced to different degrees at 48 h and/or 96 h. The activity of antioxidant enzymes (SOD, CAT, GST and GPx) and MDA content increased but GSH content decreased, indicating that ASA caused oxidative stress in D. magna. ASA also changed the expression of DNA methylation genes, such as DNMT and TET2, in D. magna. We speculated that ASA may affect the antioxidant system responses through regulation of Nrf2/Keap1 signaling pathway, and/or through indirectly influencing DNA methylation levels by DNMT and TET gene expression, but the detailed mechanism needs further investigations. Chronic exposure to ASA for 21 days caused inhibitions on the growth, reproduction and behavior of D. magna (e.g., delaying days to the first brood and shortening the body length). In summary, ASA significantly affected the antioxidant responses of D. magna, and negatively disturbed its life traits in growth, development and reproduction. PubDate: 2023-01-21
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Abstract: Abstract This study aimed to apply the comet assay on Aegla platensis crabs as a suitable non-destructive approach for in situ monitoring of freshwater genotoxicity. Animals were captured during four sampling periods in a stream under minor anthropogenic impacts in Southern Brazil. Crabs were captured with a hand net, then the hemolymph samples were collected, and the animals were released into the stream after a 20-min recovery time. Hemolymph samples were transported to the laboratory and used to perform the alkaline comet assay. Results showed an intermediate level in the DNA damage index (range 107.3–165.0 arbitrary unit). No significant differences were observed among the different sampling periods. Hemolymph was successfully used as a non-lethal source of biological samples, and the comet assay using A. platensis proved to be a feasible approach for genotoxicity studies. PubDate: 2023-01-21
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Abstract: Abstract The use of fungicides in agriculture has been playing a role in the enhancement of agricultural yields through the control of pathogens causing serious diseases in crops. Still, adverse environmental and human health effects resulting from its application have been reported. In this study, the possibility of readjusting the formulation of a commercial product combining azoxystrobin and tebuconazole (active ingredients – AIs; Custodia®) towards environmentally safer alternative(s) was investigated. Specifically, the sensitivity of non-target aquatic communities to each AI was first evaluated by applying the Species Sensitivity Distributions (SSDs) approach. Then, mixtures of these AIs were tested in a non-target organism (Raphidocelis subcapitata) denoting sensitivity to both AIs as assessed from SSDs. The resulting data supported the design of the last stage of this study, where mixtures of those AIs at equivalent vs. alternative ratios and rates as in the commercial formulation were tested against two target fungal species: Pyrenophora teres CBS 123929 and Rhynchosporium secalis CBS 110524. The comparison between the sensitivity of non-target aquatic species and the corresponding efficacy towards target fungi revealed that currently applied mixture and rates of these AIs are generally environmentally safe (antagonistic interaction; concentrations below the EC1 for R. subcapitata and generally below the HC5 for aquatic non-target communities), but ineffective against target organisms (maximum levels of inhibition of 70 and 50% in P. teres CBS 123929 and R. secalis CBS 110524, respectively). Results additionally suggest a potentiation of the effects of the AIs by the other formulants added to the commercial product at tested rates. Overall, this study corroborates that commercial products can be optimized during design stages based on a systematic ecotoxicological testing for ingredient interactions and actual efficacy against targets. This could be a valuable pathway to reduce environmental contamination during transition to a more sustainable agricultural production. PubDate: 2023-01-18
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