Seasonal dynamics of the standard test species Lemna sp. in outdoor microcosms.

Lemna L. sp. is a free-floating aquatic macrophyte that plays a key role as a standard test species in aquatic risk assessment for herbicides and other contaminants. Population modeling can be used to extrapolate from laboratory to field conditions. However, there are insufficient data on longer-term seasonal dynamics of this species to evaluate such models. Therefore, several long-term growth experiments were conducted in outdoor microcosms (surface area 0.174 m2). Monitoring parameters included biomass, frond numbers, water parameters, and weather data. Three different datasets were generated: frond numbers and biomass from weekly to monthly destructively sampled microcosms; a year-round dataset of frond numbers from five continuously monitored microcosms; and seasonal growth rates without the effect of density dependence over 1-2 weeks in freshly inoculated microcosms. Lemna sp. reached a maximum of approximately 500 000 fronds m-2 and 190 g dry weight m-2. During the first winter, the microcosms were covered by ice for approximately four weeks, and Lemna sp. populations collapsed. The second winter was warmer, without any ice cover, and Lemna sp. populations maintained high abundance throughout the winter. Dry weight per frond was not constant throughout the year but was highest in autumn and winter. Growth rates without density dependence under outdoor environmental conditions reached 0.29 day-1 for frond number, 0.43 day-1 for fresh weight, and 0.39 day-1 for dry weight. In linear regressions, these growth rates were best explained by water temperature. For the populations continuously monitored throughout a year, the nitrogen-to-phosphorus ratio best explained the growth rate of frond numbers. This study yielded a relevant dataset for testing and refining Lemna population models used in chemical risk assessment as well as for managing ecosystems and combating the effects of eutrophication. Integr Environ Assess Manag 2024;20:1625-1638. © 2024 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Seasonal dynamics of the macrophyte test species Myriophyllum spicatum over two years in experimental ditches for population modeling application in risk assessment.

Myriophyllum spicatum is a sediment-rooted, aquatic macrophyte growing submerged, with a wide geographical distribution and high ecological relevance in freshwater ecosystems. It is used in testing and risk assessment for pesticides in water and sediment. Population models enable effects measured under laboratory conditions to be extrapolated to effects expected in the field with time-variable environmental factors including exposure. These models are a promising tool in higher-tier risk assessments. However, there is a lack of data on the seasonal dynamics of M. spicatum, which is needed to test model predictions of typical population dynamics in the field. To generate such data, a two-year study was set up in outdoor experimental systems from May 2017 to May 2019. The growth of M. spicatum was monitored in 0.2025 m2 plant baskets installed in an experimental ditch. Parameters monitored included biomass (fresh weight [FW] and dry weight [DW]), shoot length, seasonal short-term growth rates of shoots, relevant environmental parameters, and weather data. The results showed a clear seasonal pattern of biomass and shoot length and their variability. M. spicatum reached a maximum total shoot length (TSL) of 279 m m-2 and a maximum standing crop above-ground DW of 262 g m-2 . Periodical growth rates reached up to 0.072, 0.095, and 0.085 day-1 for total length, FW, and DW, respectively. Multivariate regression revealed that pH (as a surrogate for the availability of carbon species) and water temperature could explain a significant proportion of the variability in M. spicatum growth rates (p < 0.05). This study has provided an ecologically relevant data set on seasonal population dynamics representative of shallow freshwater ecosystems, which can be used to test and refine population models for use in chemical risk assessment and ecosystem management. Integr Environ Assess Manag 2022;18:1375-1386. © 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Exposure and effects of sediment-spiked fludioxonil on macroinvertebrates and zooplankton in outdoor aquatic microcosms.

Information from effects of pesticides in sediments at an ecosystem level, to validate current and proposed risk assessment procedures, is scarce. A sediment-spiked outdoor freshwater microcosm experiment was conducted with fludioxonil (lipophilic, non-systemic fungicide) to study exposure dynamics and treatment-related responses of benthic and pelagic macroinvertebrates and zooplankton. Besides blank control and solvent control systems the experiment had six different treatment levels (1.7-614mga.s./kg dry sediment) based around the reported 28-d No Observed Effect Concentration (NOEC) for Chironomus riparius (40mga.s./kg dry sediment). Twelve systems were available per treatment of which four were sacrificed on each of days 28, 56 and 84 after microcosm construction. Fludioxonil persisted in the sediment and mean measured concentrations were 53-82% of the initial concentration after 84days. The dissipation rate increased with the treatment level. Also exposure concentrations in overlying water were long-term, with highest concentrations 28days after initiation of the experiment. Sediment-dwelling Oligochaeta and pelagic Rotifera and Cladocera showed the most pronounced treatment-related declines. The most sensitive sediment-dwelling oligochaete was Dero digitata (population NOEC 14.2mga.s./kg dry sediment). The same NOEC was calculated for the sediment-dwelling macroinvertebrate community. The most sensitive zooplankton species was the cladoceran Diaphanosoma brachyurum (NOEC of 1.6µga.s./L in overlying water corresponding to 5.0mga.s./kg dry sediment). At the two highest treatments several rotifer taxa showed a pronounced decrease, while the zooplankton community-level NOEC was 5.6µga.s./L (corresponding to 14.2mga.s./kg dry sediment). Zooplankton taxa calanoid Copepoda and Daphnia gr. longispina showed a pronounced treatment-related increase (indirect effects). Consequently, an assessment factor of 10 to the chronic laboratory NOECs of Chironomus riparius (sediment) and Daphnia magna (water) results in a regulatory acceptable concentration that is sufficiently protective for both the sediment-dwelling and pelagic organisms in the microcosms.

Effects of linuron on a rooted aquatic macrophyte in sediment-dosed test systems.

Effects of linuron on the sediment-rooted aquatic macrophyte Myriophyllum spicatum L. were studied in sediment-dosed test systems following a proposed guideline with extended test duration. Sediment, pore water, overlying water and macrophyte shoots were sampled weekly for chemical analyses. Linuron was stable in the sediments. Sediment and pore water concentrations were in equilibrium after 48 h. Overlying water concentrations increased over time, but did not reach equilibrium with pore water concentrations and were 100 times lower. Mass balances showed a rapid uptake of linuron by macrophyte roots. Known pathways and the compound's properties support the conclusion that Myriophyllum takes up linuron from pore water directly through the roots. Hence, effects on macrophytes in this type of sediment toxicity test should be expressed in terms of pore water concentrations. Pore water concentration is the most relevant parameter for describing effects on macrophytes.

Spider hosts (Arachnida, Araneae) and wasp parasitoids (Insecta, Hymenoptera, Ichneumonidae, Ephialtini) matched using DNA barcodes.

The study of parasitoids and their hosts suffers from a lack of reliable taxonomic data. We use a combination of morphological characters and DNA sequences to produce taxonomic determinations that can be verified with reference to specimens in an accessible collection and DNA barcode sequences posted to the Barcode of Life database (BOLD). We demonstrate that DNA can be successfully extracted from consumed host spiders and the shed pupal case of a wasp using non-destructive methods. We found Acrodactylaquadrisculpta to be a parasitoid of Tetragnathamontana; Zatypotapercontatoria and Zatypotabohemani both are parasitoids of Neottiurabimaculata. Zatypotaanomala is a parasitoid of an as yet unidentified host in the family Dictynidae, but the host species may be possible to identify in the future as the library of reference sequences on BOLD continues to grow. The study of parasitoids and their hosts traditionally requires specialized knowledge and techniques, and accumulating data is a slow process. DNA barcoding could allow more professional and amateur naturalists to contribute data to this field of study. A publication venue dedicated to aggregating datasets of all sizes online is well suited to this model of distributed science.

Ecological impacts of time-variable exposure regimes to the fungicide azoxystrobin on freshwater communities in outdoor microcosms.

This paper evaluates the effects of different time-varying exposure patterns of the strobilurin fungicide azoxystrobin on freshwater microsocosm communities. These exposure patterns included two treatments with a similar peak but different time-weighted average (TWA) concentrations, and two treatments with similar TWA but different peak concentrations. The experiment was carried out in outdoor microcosms under four different exposure regimes; (1) a continuous application treatment of 10 µg/L (CAT(10)) for 42 days (2), a continuous application treatment of 33 µg/L (CAT(33)) for 42 days (3), a single application treatment of 33 µg/L (SAT(33)) and (4) a four application treatment of 16 µg/L (FAT(16)), with a time interval of 10 days. Mean measured 42-d TWA concentrations in the different treatments were 9.4 µg/L (CAT(10)), 32.8 µg/L (CAT(33)), 14.9 µg/L (SAT(33)) and 14.7 µg/L (FAT(16)). Multivariate analyses demonstrated significant changes in zooplankton community structure in all but the CAT(10) treated microcosms relative to that of controls. The largest adverse effects were reported for zooplankton taxa belonging to Copepoda and Cladocera. By the end of the experimental period (day 42 after treatment), community effects were of similar magnitude for the pulsed treatment regimes, although the magnitude of the initial effect was larger in the SAT(33) treatment. This indicates that for long-term effects the TWA is more important for most zooplankton species in the test system than the peak concentration. Azoxystrobin only slightly affected some species of the macroinvertebrate, phytoplankton and macrophyte assemblages. The overall no observed ecologically adverse effect concentrations (NOEAEC) in this study was 10 µg/L.

Macroinvertebrate responses to insecticide application between sprayed and adjacent nonsprayed ditch sections of different sizes.

Under typical agricultural use of an insecticide, it is likely that only part of an edge-of-field drainage ditch will be directly contaminated by spray drift. The response, including recovery, of aquatic macroinvertebrates in sprayed ditch sections may be affected by immigration of organisms from adjacent nonsprayed ditch sections, but also the population dynamics in nonsprayed sections (refuges) may be affected by nearby contaminated patches (known as action at a distance). Experimental ditches were used to study the influence of the presence of nearby refuges on the responses of macroinvertebrates in ditch sections directly sprayed with the insecticide lufenuron, and vice versa. The treatment regimes differed in the proportion of the ditch (0, 33, 67, and 100% of surface area) that was sprayed to reach a lufenuron concentration of 3 microg/L in the water column of the sprayed ditch section. In sprayed ditch sections, clear treatment-related effects were observed for adult midges in the emergence traps and for aquatic arthropods (mainly juveniles) in the artificial substrate/sweep net samples. The extent in magnitude and duration of effects in sprayed ditch sections was overall larger when a larger proportion of the ditch was sprayed and/or the distance to the refuge was larger. In nonsprayed ditch sections of partially treated ditches, treatment-related effects were absent or minor for macroinvertebrates that predominantly dwell on or in the sediment compartment, particularly at a larger distance from the sprayed ditch sections. More mobile arthropods that predominantly dwell in the water column showed clear treatment-related effects in the nonsprayed ditch sections as well, but action at a distance was smaller if a smaller proportion of ditch was treated.

Impact of a benzoyl urea insecticide on aquatic macroinvertebrates in ditch mesocosms with and without non-sprayed sections.

The long-term response, including recovery, of aquatic macroinvertebrates to short-term insecticide exposure may be affected by the presence of uncontaminated refuges in the stressed ecosystem. Experimental ditches were used to study the influence of non-sprayed ditch sections regarding the ecotoxicological effects on and the recovery of macroinvertebrates following treatment with the insecticide lufenuron. The treatment regimes differed in the proportion of the ditch (0, 33, 67, and 100% of surface area) that was sprayed to reach a lufenuron concentration of 3 microg/L in the water column of the sprayed ditch section. The magnitude and duration of effects on macroinvertebrates, and on arthropods in particular, were higher when a larger proportion of the ditch was treated. Initially, more pronounced responses were observed for bivoltine and multivoltine insects and macrocrustaceans than for univoltine and semivoltine arthropods. Most macroinvertebrate arthropods showed delayed responses, with maximum treatment-related effects observed two to six weeks after lufenuron application. This latency of effects can be explained by the mode of action of lufenuron, involving inhibition of chitin synthesis, which affects arthropod molting and metamorphosis. The observed effects were short-lived only in those ditches where 33% of the surface area was sprayed. In the ditches where 67 and 100% of the surface area was sprayed, some insects and macrocrustaceans showed long-term effects. In the 100% sprayed ditches in particular, the treatment-related reduction in arthropods resulted in indirect effects, such as an increase in snails, and later in an increase in the ephemeropteran Cloeon dipterum, probably because of an increase in periphyton, and release from competition and predation. Effects that are most likely indirect also were observed for Oligochaeta, Hirudinea, and the flatworm Mesostoma sp.

Sensitivity of submersed freshwater macrophytes and endpoints in laboratory toxicity tests.

The toxicological sensitivity and variability of a range of macrophyte endpoints were statistically tested with data from chronic, non-axenic, macrophyte toxicity tests. Five submersed freshwater macrophytes, four pesticides/biocides and 13 endpoints were included in the statistical analyses. Root endpoints, reflecting root growth, were most sensitive in the toxicity tests, while endpoints relating to biomass, growth and shoot length were less sensitive. The endpoints with the lowest coefficients of variation were not necessarily the endpoints, which were toxicologically most sensitive. Differences in sensitivity were in the range of 10-1000 for different macrophyte-specific endpoints. No macrophyte species was consistently the most sensitive. Criteria to select endpoints in macrophyte toxicity tests should include toxicological sensitivity, variance and ecological relevance. Hence, macrophyte toxicity tests should comprise an array of endpoints, including very sensitive endpoints like those relating to root growth.

Ecological impact in ditch mesocosms of simulated spray drift from a crop protection program for potatoes.

Outdoor aquatic ditch mesocosms were treated with a range of pesticides to simulate various spray drift rates resulting from a typical crop protection program used in the cultivation of potatoes in The Netherlands. The main experimental aims of the present study were to provide information on the fate and ecological effects of drift of the pesticides into surface water and to evaluate the effectiveness of drift-reduction measures in mitigating risks. The pesticides selected and the dosage, frequency, and timing of application were based on normal agricultural practices in the potato crop. Applications of prosulfocarb, metribuzin (both herbicides), lambda-cyhalothrin (insecticide), chlorothalonil, and fluazinam (both fungicides) were made in the sequence typical of the spray calendar for potatoes. A total of 15 treatments with the various compounds were made by spray application to the water surface at 0.2%, 1%, and 5% of the recommended label rates. Chemical fate and effects on ecosystem function and structure (phytoplankton, zooplankton, chlorophyll-a, macroinvertebrates, macrophytes, breakdown of plant litter) were investigated. To interpret the observed effects, treatment concentrations were also expressed in toxic units (TU), which describe the relative toxicity of the compounds with standard toxicity test organisms (Daphnia and algae). After treatment, each compound disappeared from the water phase within 2 d, with the exception of prosulfocarb, for which 50% dissipation time (DT50) values ranged between 6 and 7 d. At the 5% treatment level, an exposure peak of 0.9 TUalgae was observed, which resulted in short-term responses of pH, oxygen, and phytoplankton. At the 5% treatment level, exposure concentrations also exceeded 0.1 TUDaphnia, and this resulted in long-term effects on zooplankton and macroinvertebrates, some of which did not fully recover by the end of the present study. At the 1% treatment level, only slight transient effects were observed on a limited number of zooplankton and macro-invertebrate species and on pH. At the 0.2% level, no consistent treatment-related effects were observed. Most of the observed effects were consistent with the results from higher-tier and mesocosm studies with the individual compounds. Multi and repeated stress played a small role within the applied pesticide package, because of rapid dissipation of most substances and the absence of many simultaneous applications. This suggests that risk assessments based on the individual compounds would in this case have been sufficiently protective for their uses in a crop protection program.

Effects of lambda-cyhalothrin in two ditch microcosm systems of different trophic status.

The fate and effects of the pyrethroid insecticide lambda-cyhalothrin were compared in mesotrophic (macrophyte-dominated) and eutrophic (phytoplankton-dominated) ditch microcosms (approximately 0.5 m3). Lambda-cyhalothrin was applied three times at one-week intervals at concentrations of 10, 25, 50, 100, and 250 ng/L. The rate of dissipation of lambda-cyhalothrin in the water column of the two types of test systems was similar. After 1 d, only 30% of the amount applied remained in the water phase. Initial, direct effects were observed primarily on arthropod taxa. The most sensitive species was the phantom midge (Chaoborus obscuripes). Threshold levels for slight and transient direct toxic effects were similar (10 ng/L) between types of test systems. At treatment levels of 25 ng/L and higher, apparent population and community responses occurred. At treatments of 100 and 250 ng/L, the rate of recovery of the macroinvertebrate community was lower in the macrophyte-dominated systems, primarily because of a prolonged decline of the amphipod Gammarus pulex. This species occurred at high densities only in the macrophyte-dominated enclosures. Indirect effects (e.g., increase of rotifers and microcrustaceans) were more pronounced in the plankton-dominated test systems, particularly at treatment levels of 25 ng/L and higher.