Pet dogs transfer veterinary medicines to the environment.

Worldwide, the number of pet dogs increases yearly, and as a result so does the use of veterinary medicines for flea and tick control. We investigated the potential transfer of veterinary flea products from dogs to the environment in a 'proof of principle' experiment. For this purpose, samples of hair, urine, and water after swimming were investigated. Nine dogs were recruited for this study, eight of which had been recently treated with an ectoparasiticide product. Hair and urine samples were tested for afoxalaner, fluralaner, fipronil and imidacloprid. Interestingly, contamination with ectoparasiticides was frequently demonstrated in samples from dogs untreated with these particular substances, suggesting widespread secondary transfer. In addition, hair retrieved from a bird's nest contained fipronil, fluralaner and imidacloprid, indicating a potential pathway for the exposure of juvenile birds. Three of the dogs also participated in a swimming experiment. One had been treated with oral fluralaner, whilst the remaining two had received other compounds not included in our study. However, in all three dogs, both fluralaner and imidacloprid were detected in hair samples. Fluralaner concentrations in the swimming water exceeded Dutch water quality standards, indicating a potential risk to the aquatic environment. Imidacloprid levels increased after each swimming dog, but did not breach Dutch water quality standard levels. These findings all call for improvements in the current risk assessment and management for veterinary medicines, by including companion animals and their exposure pathways into ecosystems.

Response of a nematode community to the fungicide fludioxonil in sediments of outdoor freshwater microcosms compared to a single species toxicity test.

When entering aquatic ecosystems, hydrophobic organic chemicals like the fungicide fludioxonil partition to the sediment compartment where they pose potential risks to benthic invertebrates. To assess the ecological risk for sediment-dwelling invertebrates, nematodes are a suitable organism group, as they are abundantly present and possess key positions in the benthic food web. Therefore, the toxicity of the fungicide fludioxonil to nematodes was assessed in a standardized sediment toxicity test with Caenorhabditis elegans (ISO 10872), as well as in an outdoor sediment-spiked microcosm test system. In the microcosms, effects on the nematode species composition were studied, while exposure concentrations of fludioxonil were monitored in total sediment and pore water. Toxic effects on nematodes were better predicted using concentrations in pore water than total sediment concentrations. In laboratory single species tests, fludioxonil showed considerably lower toxicity in spiked field-collected sediment, compared to artificial ISO-sediments. Applying an assessment factor of 10 to the C. elegans 96-h EC10, a Tier-1 RACNematode of 7.99 mg kg-1 dry artificial sediment (corresponding to 69 µg l-1 in pore water) appeared to be protective for nematode communities in microcosms that showed no response in total abundance and species composition up to 39.9 mg fludioxonil kg-1 dry sediment (corresponding to 392 µg l-1 in pore water).

Toxicity of sediment-bound lufenuron to benthic arthropods in laboratory bioassays.

This paper deals with species sensitivity distributions (SSDs) for the lipophilic insecticide lufenuron and benthic arthropods based on sediment-spiked laboratory toxicity tests. This compound that inhibits chitin synthesis and moulting of arthropods persists in sediment. Using field-collected sediment, toxicity tests were conducted with three macro-crustaceans and six insects. The Hazardous Concentration to 5% of the tested species, the HC5 (and 95% confidence limit), derived from an SSD constructed with 10d-LC50's was 2.2 (1.2-5.7) µg/g organic carbon (OC) in dry sediment. In addition, HC5 values derived from SSDs constructed with 28d-LC10 and 28-d LC50 values were 0.13 (0.02-1.50) µg/g OC and 2.0 (1.3-5.5) µg/g OC, respectively. In 28d toxicity tests with Chironomus riparius and Hyalella azteca, a higher sensitivity was observed when using lufenuron-spiked field-collected sediment than in lufenuron-spiked artificial sediment. Overall, the non-biting midge C. riparius appeared to be a representative and sensitive standard test species to assess effects of lufenuron exposure in sediment. The Tier-1 (based on standard test species), Tier-2 (based on standard and additional test species) and Tier-3 (model ecosystem approach) regulatory acceptable concentrations (RACs) for sediment-spiked lufenuron did not differ substantially. The Tier-2 RAC was the lowest. Since to our knowledge this study is the first in the open literature that evaluates the tiered approach in the sediment effect assessment procedure for pesticides, we advocate that similar evaluations should be conducted for pesticides that differ in toxic mode-of-action.

Effects of sediment-spiked lufenuron on benthic macroinvertebrates in outdoor microcosms and single-species toxicity tests.

Sediment ecotoxicity studies were conducted with lufenuron to (i) complement the results of a water-spiked mesocosm experiment with this lipophilic benzoylurea insecticide, (ii) to explore the predictive value of laboratory single-species tests for population and community-level responses of benthic macroinvertebrates, and (iii) to calibrate the tier-1 effect assessment procedure for sediment organisms. For this purpose the concentration-response relationships for macroinvertebrates between sediment-spiked microcosms and those of 28-d sediment-spiked single-species toxicity tests with Chironomus riparius, Hyalella azteca and Lumbriculus variegatus were compared. Lufenuron persisted in the sediment of the microcosms. On average, 87.7% of the initial lufenuron concentration could still be detected in the sediment after 12 weeks. Overall, benthic insects and crustaceans showed treatment-related declines and oligochaetes treatment-related increases. The lowest population-level NOEC in the microcosms was 0.79µg lufenuron/g organic carbon in dry sediment (µg a.s./g OC) for Tanytarsini, Chironomini and Dero sp. Multivariate analysis of the responses of benthic macroinvertebrates revealed a community-level NOEC of 0.79µg a.s./g OC. The treatment-related responses observed in the microcosms are in accordance with the results of the 28-d laboratory toxicity tests. These tests showed that the insect C. riparius and the crustacean H. azteca were approximately two orders of magnitude more sensitive than the oligochaete L. variegatus. In our laboratory tests, using field-collected sediment, the lowest 28-d EC10 (0.49µg a.s./g OC) was observed for C. riparius (endpoint survival), while for the standard OECD test with this species, using artificial sediment, a NOEC of 2.35µg a.s./g OC (endpoint emergence) is reported. In this particular case, the sediment tier-1 effect assessment using the chronic EC10 (field-collected sediment) or chronic NOEC (artificial sediment) of C. riparius and an assessment factor of 10, seems to be protective for the treatment-related responses observed in the sediment-spiked microcosms.

The minimum detectable difference (MDD) and the interpretation of treatment-related effects of pesticides in experimental ecosystems.

In the European registration procedure for pesticides, microcosm and mesocosm studies are the highest aquatic experimental tier to assess their environmental effects. Evaluations of microcosm/mesocosm studies rely heavily on no observed effect concentrations (NOECs) calculated for different population-level endpoints. Ideally, a power analysis should be reported for the concentration-response relationships underlying these NOECs, as well as for measurement endpoints for which significant effects cannot be demonstrated. An indication of this statistical power can be provided a posteriori by calculated minimum detectable differences (MDDs). The MDD defines the difference between the means of a treatment and the control that must exist to detect a statistically significant effect. The aim of this paper is to expand on the Aquatic Guidance Document recently published by the European Food Safety Authority (EFSA) and to propose a procedure to report and evaluate NOECs and related MDDs in a harmonised way. In addition, decision schemes are provided on how MDDs can be used to assess the reliability of microcosm/mesocosm studies and for the derivation of effect classes used to derive regulatory acceptable concentrations. Furthermore, examples are presented to show how MDDs can be reduced by optimising experimental design and sampling techniques.

Survey of the crayfish plague pathogen presence in the Netherlands reveals a new Aphanomyces astaci carrier.

North American crayfish species as hosts for the crayfish plague pathogen Aphanomyces astaci contribute to the decline of native European crayfish populations. At least six American crayfish species have been reported in the Netherlands but the presence of this pathogenic oomycete with substantial conservational impact has not yet been confirmed in the country. We evaluated A. astaci prevalence in Dutch populations of six alien crustaceans using species-specific quantitative PCR. These included three confirmed crayfish carriers (Orconectes limosus, Pacifastacus leniusculus, Procambarus clarkii), two recently introduced but yet unstudied crayfish (Orconectes cf. virilis, Procambarus cf. acutus), and a catadromous crab Eriocheir sinensis. Moderate levels of infection were observed in some populations of O. limosus and P. leniusculus. Positive results were also obtained for E. sinensis and two Dutch populations of O. cf. virilis. English population of the latter species was also found infected, confirming this taxon as another A. astaci carrier in European waters. In contrast, Dutch P. clarkii seem only sporadically infected, and the pathogen was not yet detected in P. cf. acutus. Our study is the first confirmation of crayfish plague infections in the Netherlands and demonstrates substantial variation in A. astaci prevalence among potential hosts within a single region, a pattern possibly linked to their introduction history and coexistence.

In situ treatment with activated carbon reduces bioaccumulation in aquatic food chains.

In situ activated carbon (AC) amendment is a new direction in contaminated sediment management, yet its effectiveness and safety have never been tested on the level of entire food chains including fish. Here we tested the effects of three different AC treatments on hydrophobic organic chemical (HOC) concentrations in pore water, benthic invertebrates, zooplankton, and fish (Leuciscus idus melanotus). AC treatments were mixing with powdered AC (PAC), mixing with granular AC (GAC), and addition-removal of GAC (sediment stripping). The AC treatments resulted in a significant decrease in HOC concentrations in pore water, benthic invertebrates, zooplankton, macrophytes, and fish. In 6 months, PAC treatment caused a reduction of accumulation of polychlorobiphenyls (PCB) in fish by a factor of 20, bringing pollutant levels below toxic thresholds. All AC treatments supported growth of fish, but growth was inhibited in the PAC treatment, which was likely explained by reduced nutrient concentrations, resulting in lower zooplankton (i.e., food) densities for the fish. PAC treatment may be advised for sites where immediate ecosystem protection is required. GAC treatment may be equally effective in the longer term and may be adequate for vulnerable ecosystems where longer-term protection suffices.

Copper kinetics and internal distribution in the marbled crayfish (Procambarus sp.).

Metal pollution e.g. copper, in water bodies occurs worldwide. Although copper is an essential trace metal, at certain levels it is still considered as pollutant. The aim of this study was to investigate the effect of exposure concentration on copper bioaccumulation in marbled crayfish (Procambarus sp.) by determining uptake and elimination kinetics. Crayfish were exposed to sub-lethal copper concentrations (average measured concentrations of 0.031 and 0.38 mg Cu L(-1)) for 14 d and transferred to copper-free water for another 14 d. At different time points during the uptake and elimination phases copper concentrations were measured in five organs (exoskeleton, gills, muscle, ovaries and hepatopancreas). At 0.031 mg Cu L(-1), copper levels in the crayfish organs were not significantly increased compared to the control animals, suggesting effective regulation. Exposure to 0.38 mg Cu L(-1) did lead to not significantly increased copper levels in muscles and ovaries, while the gills and exoskeleton, which are in direct contact with the water, showed significantly higher copper concentrations. In these four organs, copper showed fast uptake kinetics with equilibrium reached within 10 d of exposure. Copper accumulation was highest in the hepatopancreas; uptake in this storage organ steadily increased with time and did not reach equilibrium within the 14-d exposure period. Copper accumulation levels in the marbled crayfish found in this study were hepatopancreas>gills>exoskeleton>muscle.

The species sensitivity distribution approach compared to a microcosm study: a case study with the fungicide fluazinam.

We assessed the sensitivity of freshwater organisms (invertebrates and algae) to the fungicide Shirlan (active ingredient fluazinam) in single-species laboratory tests and in microcosms. Species sensitivity distribution (SSD) curves were constructed by means of acute toxicity data for 14 invertebrate species, since algae were much less sensitive. The EC(10)-based SSD gave a median HC(5) value of 0.6microgL(-1) and a 90% confidence interval of 0.1-1.9 microgL(-1). The EC(50)-based SSD gave a median HC(5) value of 3.9 microgL(-1) (90% confidence interval: 0.9-9.9 microgL(-1)). The microcosms were treated four times with Shirlan (concentration range: 0.4-250 microgL(-1)). Responses of the microcosm communities were followed. The 2 microgL(-1) treatment was the no-observed-effect concentration (NOEC(microcosm)). The 10 microgL(-1) treatment resulted in short-term effects on a few zooplankton taxa. Clear effects were observed at 50 and 250 microgL(-1). The responses in the microcosms were in line with the toxicity data for the tested lab species. The median EC(10)-based HC(5) and the lower limit EC(50)-based HC(5) were lower, and the median EC(50)-based HC(5) was slightly higher than the NOEC(microcosm). This is consistent with other studies that compared SSDs with responses in model ecosystems that received repeated applications of pesticides.

Impact of triphenyltin acetate in microcosms simulating floodplain lakes. II. Comparison of species sensitivity distributions between laboratory and semi-field.

The study objectives were to shed light on the types of freshwater organism that are sensitive to triphenyltin acetate (TPT) and to compare the laboratory and microcosm sensitivities of the invertebrate community. The responses of a wide array of freshwater taxa (including invertebrates, phytoplankton and macrophytes) from acute laboratory Single Species Tests (SST) were compared with the concentration-response relationships of aquatic populations in two types of freshwater microcosms. Representatives of several taxonomic groups of invertebrates, and several phytoplankton and vascular plant species proved to be sensitive to TPT, illustrating its diverse modes of toxic action. Statistically calculated ecological risk thresholds (HC5 values) based on 96 h laboratory EC50 values for invertebrates were 1.3 microg/l, while these values on the basis of microcosm-Species Sensitivity Distributions (SSD) for invertebrates in sampling weeks 2-8 after TPT treatment ranged from 0.2 to 0.6 microg/l based on nominal peak concentrations. Responses observed in the microcosms did not differ between system types and sampling dates, indicating that ecological threshold levels are not affected by different community structures including taxa sensitive to TPT. The laboratory-derived invertebrate SSD curve was less sensitive than the curves from the microcosms. Possible explanations for the more sensitive field response are delayed effects and/or additional chronic exposure via the food chain in the microcosms.

Impact of triphenyltin acetate in microcosms simulating floodplain lakes. I. Influence of sediment quality.

Floodplain lakes in the Rhine-Meuse delta of the Netherlands vary considerably in levels of sediment-bound toxicants. Microcosm experiments were done to compare the ecological impact of the fungicide triphenyltin acetate (TPT) between test systems with clean or polluted sediments (10 microcosms each). Differences in sediment quality affected the structure of the aquatic communities that developed in the microcosms. Initially, a faster growth of the macrophyte Elodea nuttallii was observed on the polluted sediments, which contained not only toxicants but also higher organic matter and nutrient levels. Dynamics of TPT concentrations in the overlying water were very similar between the two types of test system. Higher levels of TPT, however, were found in the sediment compartment of the clean sediment systems containing a smaller macrophyte biomass. TPT was very persistent in the sediments. In both test systems representatives of several taxonomic groups showed clear responses to a single application of TPT, although benthic Nematoda were not affected. Although a few differences in the intensity and/or duration of TPT-related population responses were observed between the two types of test system, the background pollutants in the polluted sediment hardly affected the overall sensitivity of the aquatic community to the additional chemical stressor TPT.

Life-history consequences for Daphnia pulex exposed to pharmaceutical carbamazepine.

The effects of the antiepileptic, analgesic drug carbamazepine on the growth, morphology, and life-history characteristics of Daphnia pulex were examined at nominal concentrations of 0, 0.1, 1, 10, 100, and 200 microg L(-1). At 1 microg carbamazepine L(-1), Daphnia matured and reproduced slightly earlier than did controls, and at a given body length females produced more offspring than did controls or those receiving other treatments. In combination with a relatively high juvenile somatic growth rate and highest total number of progeny produced per female, carbamazepine at 1 microg L(-1) seemed to exert a stimulatory effect. The rates of population growth of the 100 and 200 microg L(-1) treatment groups was 9% and 32% lower, respectively, than the rates of growth of the controls and the Daphnia receiving treatments of up to 10 microg carbamazepine L(-1). At the highest dose, retardation of juvenile somatic growth resulted in delayed maturity and consequently in a lower rate of population growth. Adult somatic growth, spine length, reproductive output, and size of newborns were similar among treatments. Male offspring were only produced in the third broods, with broods that were 8% and 28% male at 1 and 10 microg L(-1), respectively. Neck teeth were never observed in Daphnia. Chronic adverse effects of carbamazepine on nontarget Daphnia were detected at 200 microg carbamazepine L(-1), but stimulatory effects might occur at environmentally realistic concentrations. However, additional studies of chronic toxicity investigating various combinations of pharmaceuticals and various environmental stresses, such as food condition, temperature, and kairomones, are needed to fully explore potential long-term adverse effects and to assess the environmental risk of common pharmaceuticals.