The Survival Response of Earthworm (Eisenia fetida L.) to Individual and Binary Mixtures of Herbicides.

Frequent use of herbicides may impose a risk on non-target species. The objective was to test the combined toxic effect of binary herbicide mixtures­metribuzin:halosulfuron and metribuzin:flumioxazin­on non-target earthworms in two test systems: filter paper and a soil toxicity test system. The joint action experiments were independently run twice to substantiate the findings. The most potent individual herbicide was metribuzin, with a 50% lethal concentration (LC50) of 17.17 µg ai. cm−2 at 48 h in the filter paper test. The toxicity of the individual herbicides on the filter paper test was ranked as metribuzin>halosulfuron>flumioxazin. In the soil test, metribuzin and halosulfuron had high toxicity with an LC50 of 8.48 and 10.08 mg ai. kg−1, respectively, on day 14. Thus, the individual herbicide ranking did not change between the filter paper and artificial soil tests. The herbicide's mixed effect in both test systems showed a consistent antagonistic effect relative to a Concentration Addition reference model. It indicates that the mixtures retracted the herbicide's action in the earthworms.

Assessing herbicide symptoms by using a logarithmic field sprayer.

BACKGROUND: In field experiments, assessment of herbicide selectivity and efficacy rarely takes advantage of dose-response regressions. The objective is to demonstrate that logarithmic sprayers, which automatically make a logarithmic dilution of a herbicide rate, can extract biologically relevant parameters describing the efficacy of herbicides in crops, and compare localities and time of assessment. RESULTS: In a conventional and an organic field, canola, white mustard, and no crop plots were sprayed with diflufenican and beflubutamid. A mixed effect log-logistic dose-response regression, with autoregressive correlation structure, estimated ED50 and ED90 for visual and Excess Green Index symptoms at various days after treatment (DAT). For visual assessment, ED50 differed within no crop between locations for beflubutamid at 12 DAT and 26 DAT. For diflufenican, the ED50 was different within crops at the two fields at 12 DAT, but not at 26 DAT. The Excess Green Indices at ED50 were not different among herbicides, locations, and corps; ED90 differed for white mustard and canola for beflubutamid but not for diflufenican. CONCLUSION: Suitable nonlinear regression models are now available for fitting dose-response data from a logarithmic sprayer in field experiments. The derived parameters (e.g. ED50 ) can compare selectivity and efficacy at numerous cropping systems. © 2018 Society of Chemical Industry.

Reproducibility of binary-mixture toxicity studies.

Binary-mixture studies often are conducted with the aim of elucidating the effect of one specific chemical on the biological action of another. The results can be interpreted in relation to reference models by the use of response-surface analyses and isobolograms. The amount of data needed for these analyses is, however, extensive, and the experiments therefore rarely are repeated. In the present study, we investigate the reproducibility of isobole shapes of binary-mixture toxicity experiments in terms of deviation from the reference model of concentration addition (CA), dose-level dependence, and isobole asymmetry. We use data from four herbicide mixtures tested in three to five independent experiments on the aquatic test plant Lemna minor and the terrestrial plant Tripleurospermum inodorum. The results showed that the variation both within and among experiments was approximately half the size for the aquatic test system compared to the terrestrial system. As a consequence, a consistent deviation from CA could be obtained in three of four herbicide mixtures for L. minor, whereas this was only the case for one or two of the herbicide mixtures tested on T. inodorum. For one mixture on T. inodorum, both CA synergism and antagonism were detected. Dose-dependent effects could not be repeated consistently, just as the asymmetry found in some isoboles could not. The study emphasizes the importance of repeating mixture toxicity experiments, especially for test systems with large variability, and using caution when drawing biological conclusions from the test results.

Activities of mixtures of soil-applied herbicides with different molecular targets.

The joint action of soil-applied herbicide mixtures with similar or different modes of action has been assessed by using the additive dose model (ADM). The herbicides chlorsulfuron, metsulfuron-methyl, pendimethalin and pretilachlor, applied either singly or in binary mixtures, were used on rice (Oryza sativa L.). The growth (shoot) response curves were described by a logistic dose-response model. The ED50 values and their corresponding standard errors obtained from the response curves were used to test statistically if the shape of the isoboles differed from the reference model (ADM). Results showed that mixtures of herbicides with similar molecular targets, i.e. chlorsulfuron and metsulfuron (acetolactate synthase (ALS) inhibitors), and with different molecular targets, i.e. pendimethalin (microtubule assembly inhibitor) and pretilachlor (very long chain fatty acids (VLCFAs) inhibitor), followed the ADM. Mixing herbicides with different molecular targets gave different results depending on whether pretilachlor or pendimethalin was involved. In general, mixtures of pretilachlor and sulfonylureas showed synergistic interactions, whereas mixtures of pendimethalin and sulfonylureas exhibited either antagonistic or additive activities. Hence, there is a large potential for both increasing the specificity of herbicides by using mixtures and lowering the total dose for weed control, while at the same time delaying the development of herbicide resistance by using mixtures with different molecular targets.

Does the effect of herbicide pulse exposure on aquatic plants depend on Kow or mode of action?

The highest concentrations of herbicides measured in flowing surface waters are often only present for short periods of time. These herbicide pulses can reach concentrations that would affect aquatic plants if present over a long time. The aim of this study was to assess the effect of a 3-h herbicide pulse relative to the effects of long-term (4 and 7 days) exposure of six herbicides with different sites of action and different K(ow) on the growth of the floating macrophyte Lemna minor. The herbicides were the two photosynthetic inhibitors: diquat and terbuthylazine, the inhibitors of acetolactate syntase (ALS), imazamox and metsulfuron-methyl and the microtubule assembly inhibitors propyzamide and pendimethalin. The log K(ow) ranged from -4.6 to 5.2. For imazamox, metsulfuron-methyl, propyzamide and pendimethalin a 3-h pulse induced the effect on area-specific growth as did a 4-day exposure at an approximate 10-fold higher concentration. For diquat and terbuthylazine a concentration closer to a factor of 100 or more was needed for a 3-h pulse to induce an effect similar to that of a 4-day exposure. For diquat, the low pulse-effect was most likely due to a slow uptake of the hydrophilic ion (log K(ow) = -4.6), as no effect was observed on chlorophyll fluorescence within 8 h after exposure. The chlorophyll fluorescence parameters are expected to respond quickly to a PSI inhibitor as diquat. For terbuthylazine, fluorescence measurements showed an effect on photosynthesis within 1h of exposure, and reached a minimum after 3 h. Recovery was fast, and initial fluorescence was restored within 24 h. Hence, the small pulse effect on area-specific growth was due to rapid recovery of photosynthesis. In contrast to terbuthylazine, the stop in area-specific growth observed for the ALS-and microtubule assembly inhibitors, took up to 4 days to recover from. Such a long recovery time after a pulse of only 3 h indicate that at realistic pulse exposures of up to a day or two, pulse-effects will approach the effects obtained in long-term studies. When investigating the effects of pulse exposures on aquatic plants, we should therefore focus more on non-photosynthetic inhibitors, which might not appear in pulses in as large concentrations as the PSII inhibitors investigated up till now, but whose effect, even in a shorter pulse, can be more damaging.

Improved empirical models describing hormesis.

During the past two decades, the phenomenon of hormesis has gained increased recognition. To promote research in hormesis, a sound statistical quantification of important parameters, such as the level and significance of the increase in response and the range of concentration where it occurs, is strongly needed. Here, we present an improved statistical model to describe hormetic dose-response curves and test for the presence of hormesis. Using the delta method and freely available software, any percentage effect dose or concentration can be derived with its associated standard errors. Likewise, the maximal response can be extracted and the growth stimulation calculated. The new model was tested on macrophyte data from multiple-species experiments and on laboratory data of Lemna minor. For the 51 curves tested, significant hormesis was detected in 18 curves, and for another 17 curves, the hormesis model described that data better than the logistic model did. The increase in response ranged from 5 to 109%. The growth stimulation occurred at an average dose somewhere between zero and concentrations corresponding to approximately 20 to 25% of the median effective concentration (EC50). Testing the same data with the hormesis model proposed by Brain and Cousens in 1989, we found no significant hormesis. Consequently, the new model is shown to be far more robust than previous models, both in terms of variation in data and in terms of describing hormetic effects ranging from small effects of a 10% increase in response up to effects of an almost 100% increase in response.

Re-evaluation of groundwater monitoring data for glyphosate and bentazone by taking detection limits into account.

Current regulatory assessment of pesticide contamination of Danish groundwater is exclusively based on samples with pesticide concentrations above detection limit. Here we demonstrate that a realistic quantification of pesticide contamination requires the inclusion of "non-detect" samples i.e. samples with concentrations below the detection limit, as left-censored observations. The median calculated pesticide concentrations are shown to be reduced 10(4) to 10(5) fold for two representative herbicides (glyphosate and bentazone) relative to the median concentrations based upon observations above detection limits alone.