Low field-relevant tebufenozide concentrations affect reproduction in Chironomus riparius (Diptera: Chironomidae) in a long-term toxicity test

A few studies reporting the effects of tebufenozide, a non-steroidal ecdysone agonist that mimics natural moulting hormones in chironomids exist in the literature. However, nothing is known about its chronic or multigenerational effects on the reproduction of aquatic insects, although tebufenozide is present in aquatic ecosystems. Here, we investigated the chronic toxicity of tebufenozide in two successive generations of Chironomus riparius using nominal concentrations that ranged from 4 to 26.2 μg/L. We started the test from the first instar larvae in the parental (P) generation, quantifying life cycle parameters (emergence, sex ratio, development rate, fecundity and fertility) until the emergence in the subsequent F1 generation. Results showed a reduction in reproduction and a significant decrease in male developmental rate of midges for all treatments, in the F1 generation compared with the P generation (paired t?test; p?<?0.001). Two-way analysis of variance revealed a significant exposure?×?generation effect on male fraction with male fraction increasing (P generation) or decreasing (F1 generation) with increasing exposure. These effects on C. riparius underline the importance of conducting long-term studies with environmentally relevant concentrations to investigate population-level endpoints for endocrine disrupting chemicals.     

An assessment of the chronic toxicity of the synthetic pyrethroid, fenvalerate, to Daphnia galeata mendotae, using life tables

The effects of chronic exposure to fenvalerate, a synthetic pyrethroid insecticide, on the cladoceran Daphnia galeata mendotae were evaluated under laboratory conditions. The daphnids were exposed for their entire life cycle to concentrations ranging from 0.005 to 0.1 microg fenvalerate litre(-1). The parameters used to determined toxicity were survival, time to first reproduction, reproductive frequency, number of young per brood, cumulative brood size, intrinsic rate of natural increase (r(m)), generation time (T) and net reproductive rate (R(0)). A concentration of 0.005 microg fenvalerate litre(-1) resulted in an increase in longevity but a decrease in production of young. Higher concentrations caused a decrease in survival. Studies of shorter duration than the complete life cycle of the organisms would not have detected toxicity at such low levels. The intrinsic rate of natural increase, r(m), was not affected by fenvalerate until concentration reached 0.05 microg litre(-1) and r(m) decreased to 0.113 from 0.238. A concentration of 0.01 microg litre(-1) caused the net reproductive rate, R(0), and the generation time, T, to decrease to 73.2 offspring per female and 17.3 days from 125.9 offspring per female and 20.3 days, respectively.     

Sperm count in earthworms (Lumbricus terrestris) as a biomarker for environmental toxicology: effects of cadmium and chlordane

Earthworms, Lumbricus terrestris, exposed in artificial soil to sublethal concentrations of technical chlordane (6.25, 12.5, and 25 ppm) and cadmium nitrate (100, 200, and 300 ppm) exhibited significant reduction in spermatozoa from testes and seminal vesicles. The onset time of reduction varied with exposure concentration, but absolute depression in sperm count was independent of exposure concentration or exposure duration after reduction was first manifested, demonstrating a threshold effect. Earthworm sperm counts show potential as a rapid-measurement endpoint biomarker for measuring sublethal effects of chemical pollutants on reproduction.     

Are individual NOEC levels safe for mixtures? A study on mixture toxicity of brominated flame-retardants in the copepod Nitocra spinipes

In aquatic ecosystems organisms are exposed to mixtures of pollutants. Still, risk assessment focuses almost exclusively on effect characterization of individual substances. The main objective of the current study was therefore to study mixture toxicity of a common group of industrial substances, i.e., brominated flame-retardants (BFRs), in the harpacticoid copepod Nitocra spinipes. Initially, 10 BFRs with high hydrophobicity but otherwise varying chemical characteristics were selected based on multivariate chemical characterization and tested individually for effects on mortality and development using a partial life cycle test (six days) where silica gel is used as a carrier of the hydrophobic substances. Based on these findings, six of the 10 BFRs were mixed in a series of NOEC proportions (which were set to 0.008, 0.04, 0.2, 1, and five times the NOEC concentrations for each individual BFR), loaded on silica gel and tested in a full life cycle test (26 days). Significantly increased mortality was observed in N. spinipes after six and 26 days exposure at a NOEC proportion that equals the NOEC LDR value (x1) for each BFR in the mixture (p=0.0015 and p=0.0105, respectively). At the NOECx5 proportion all animals were dead. None of the other NOEC proportions caused significant negative responses related to development and reproduction. This shows that low concentrations of individual substances can cause toxicity if exposed in mixtures, which highlights the need to consider mixture toxicity to a greater extent in regulatory work.     

Exposure to pyriproxyfen (juvenile hormone agonist) does not alter maternal care and reproduction in the European earwig

Sublethal exposure to pesticides can alter the survival and reproduction of a wide range of non-target organisms. However, it remains unclear whether this exposure can alter behaviours that are often essential for long-term population dynamics and maintenance, such as parental care. In this study, we tested the effect of pyriproxyfen exposure (an insect growth regulator) on maternal care in the European earwig, an insect that is both used in pest control in pip-fruit orchards and considered a pest in stone fruit orchards. We exposed 424 females at doses either 10 times lower, equivalent or 10 times higher than normal application rates in French orchards. As maternal care can change over the weeks of family life, we exposed the earwig mothers at five different days before and after egg hatching. We then measured the expression of ten forms of maternal care towards eggs and juveniles, six non-caring behaviours, eggs and juvenile development, metabolic reserves in mothers at egg hatching and females’ production of a terminal clutch. First, our results revealed that the three tested doses of pyriproxyfen were non-lethal and confirmed that maternal care decreased throughout both pre- and post-hatching family life. However, we did not detect any effect of pyriproxyfen on maternal care and non-care behaviours, eggs and juvenile development, quantities of lipids, proteins and glycogen in mothers at egg hatching, and on the production of a future clutch. Overall, these findings suggest that the maximal doses of pyriproxyfen authorized in French orchards is likely to have limited effects on the short- and long-term maintenance of populations of the European earwig and raises fundamental questions about the nature of the link between juvenile hormone and parental care in insects.

     

Factors determining the uptake of persistent pollutants in an eel population (Anguila anguilla L.)

The distribution of persistent pollutants in an eel population from a eutrophic lake of southern Scandinavia was examined. The origin of PCBs, DDT, DDE and lindane found in the fish was most likely the atmosphere. The most important factors for uptake of the chlorinated hydrocarbons was age (exposure time), growth rate and fat content. The life cycle of the eel is unique with a stage in freshwater when energy reserves (fat stored in muscular tissue) and lipophilic pollutants are accumulated. This stage is followed by a long migration to the spawning areas in the Sargasso Sea when pollutants are released from the fat deposits. These two stages followed by a once-in-a-lifetime spawning behaviour, makes the eel especially vulnerable to persistent pollutants. The effects of persistent pollutants combined with the eel's unusual life cycle may explain the decline in the eel population in northern Europe in recent decades.     

Effects of phytosterols on zebrafish reproduction in multigeneration test

Zebrafish from mixed sex populations were exposed continuously across three generations to two phytosterol preparations both containing -sitosterol. The phytosterols were isolated from wood and soy beans. Blood vitellogenin levels and sex ratio changes were used as intermediate indicators of the reproduction failures. Both sterol preparations caused vitellogenin induction in the exposed fish. The wood sterol changed the sex ratio of the exposed fish. In generation F1, the predominant sex was male, and in generation F2 it was female. The soy sterol in the used test concentration was lethal to the exposed fish in generation F1. This multigeneration test evidenced that phytosterols containing -sitosterol disrupt the reproduction system of zebrafish by changing the sex ratios and by inducing the vitellogenin production in the exposed fish.     

Population dynamics in Daphnia magna as modified by chronic tetradifon stress

Two Daphnia magna offsprings (animals from the first and third brood) whose parentals (F0-generation) were exposed during 21 days to different tetradifon (4-chlorophenyl 2,4,5-trichlophenyl sulfone) concentrations, were transferred to a pesticide free medium during 21 days (recovery period). The algae Nannochloris oculata (5 x 10(5) cells/mL) was used as food. In this recovery study, survival, growth and reproduction (mean total young per female, mean brood size, onset of reproduction and mean number broods per female) were assessed as individual parameters and the intrinsic rate of natural increase (r) as population parameter, for F1 generation (1st and 3rd broods). Reproduction was still reduced in F1 (1st and 3rd broods) generation daphnids from parentals (F0) exposed to 0.18 mg/L tetradifon and higher concentrations even after 21 days in clean water. However, survival was not significantly different (p > 0.05) in those F1 offsprings from parentals pre-exposed to the acaricide. Growth was still reduced in F1 daphnids from parentals pre-exposed to 0.10-0.44 mg/L tetradifon. The intrinsic rate of natural increase (r) was still affected in F1 generation daphnids, specially in those from the third brood. Therefore, F1 generation D. magna offsprings from a parental generation (F0) previously exposed to the acaricide tetradifon were not able to recuperate completely when a recovery period of 21 days was allowed.     

Enchytraeidae Fridericia bulbosa as a new test species for soil ecotoxicity assessment

Fridericia bulbosa (Rosa, 1887) was proposed as a new test species to assess soil ecotoxicity. The acute toxicity of mercury (Hg) and bromoxynil (BX) on the mortality of Fridericia bulbosa (F. bulbosa) in OECD soil was investigated. The results suggested there were statistically significant differences (p<0.05) between controls and treatments except lower concentration of exposure to single pollutant. BX was more toxic to F. bulbosa than Hg. The 14 d LC(50) values for F. bulbosa exposed to Hg and BX were 3.87 and 2.41 mg kg(-1), respectively. The effects of mixtures with single pollutants on earthworms were observed differently. Toxicity of Hg on earthworms was significantly influenced by the concentration of BX. BX was the main contributive factor of the combined toxic effects. It can be proved that F. bulbosa is a suitable bio-indicator to measure the acute toxicity of mixed pollutants in soil and the mortality of earthworms may be considered as a valuable and sensitive biomarker to diagnose adverse effect of Hg or BX in soil environment.     

Fluoxetine effects assessment on the life cycle of aquatic invertebrates

Fluoxetine is a serotonin re-uptake inhibitor, generally used as an antidepressant. It is suspected to provoke substantial effects in the aquatic environment. This study reports the effects of fluoxetine on the life cycle of four invertebrate species, Daphnia magna, Hyalella azteca and the snail Potamopyrgus antipodarum exposed to fluoxetine spiked-water and the midge Chironomus riparius exposed to fluoxetine-spiked sediments. For D. magna, a multi-generational study was performed with exposition of newborns from exposed organisms. Effects of fluoxetine could be found at low measured concentrations (around 10mugl(-1)), especially for parthenogenetic reproduction of D. magna and P. antipodarum. For daphnids, newborns length was impacted by fluoxetine and the second generation of exposed individuals showed much more pronounced effects than the first one, with a NOEC of 8.9mugl(-1). For P. antipodarum, significant decrease of reproduction was found for concentrations around 10mugl(-1). In contrast, we found no effect on the reproduction of H. azteca but a significant effect on growth, which resulted in a NOEC of 33mugl(-1), expressed in nominal concentration. No effect on C. riparius could be found for measured concentrations up to 59.5mgkg(-1). General mechanistic energy-based models showed poor relevance for data analysis, which suggests that fluoxetine targets specific mechanisms of reproduction.     

Detecting points as developmental delay based on the life-history development and urosome deformity of the harpacticoid copepod, Tigriopus japonicus sensu lato, following exposure to benzo(a)pyrene

To identify ecotoxicological responses to an endocrine disrupter, benzo(a)pyrene, we examined the life-history of the harpacticoid copepod, Tigriopus japonicus sensu lato. Based on the life-history of copepods, survival rate of nauplii (NSR) and copepodites (CSR), copepodite emergence day (CED) and adult male emergence day (AMED), sex ratio (MER), brooding success rate (BSR), and first brooding day of adult females (FBD) were measured. Significant differences were observed in the survival and development of nauplii (NSR and CED) and sex ratio (MER) of exposed and non-exposed copepods. Moreover, high concentration of BaP can be lethal to copepodite and exhibited a delay of growth. In this study, the CED and AMED among ecotoxicological response based on life-history developments were delayed and the body characteristics decreased in response to exposure to benzo(a)pyrene. The dwarfism and urosome deformity of the T. japonicus s.l. was exhibited in response to chemical exposure. Specifically, the body characteristics and biomass of dwarf copepods that had been exposed to benzo(a)pyrene were 30% and 50% lower than the control group, respectively. The incidence of abnormal urosomes was divided into two types. The first deformity type was signs of shrinkage in the middle of the urosome or the entire urosome was narrower than those of the control organisms. In the second type, the anal somite and the distal side of the urosome had abnormally swelled. Taken together, the nauplii and copepodid development of T. japonicus s.l. can be used as a useful biomaker for detecting developmental delay based on their entire life-history. In addition, the urosome deformity was used a good potential monitoring tool invading various chemicals and environmental contamination into water system.     

Effects of long- and short-term exposure to carbaryl on survival, growth and reproduction of Daphnia ambigua

In laboratory experiments a cladoceran, Daphnia ambigua, was exposed for six instars to the insecticide carbaryl at five different concentrations (1, 2, 3, 4, 5 microg litre(-1)) or received short-term (10-h) exposure to 5 microg carbaryl litre(-1) at different life stages of the animal. In the long-term exposure experiments carbaryl at 1 microg litre(-1) did not have any detectable effects on the animals, whereas 2 microg litre(-1) reduced the survival, growth and reproduction of the animals, and higher concentrations killed all the animals examined before they grew to the fifth instar. These results indicate that the range of sublethal concentrations of carbaryl for D. ambigua is narrow. In the short-term exposure experiments the animals did not suffer any damage when they were exposed to the chemical during the egg stage. However, their growth and egg production were reduced when the short-term exposure was conducted during the life stages from the final embryo stage to the third instar. The effect was most marked when the animals were treated during the first instar, indicating that this is the stage when the animals are most sensitive to the chemical.     

Toxicity of the herbicide glyphosate to Chordodes nobilii (Gordiida, Nematomorpha)

Nematomorpha (horsehair worms) is a poorly known group of worm-like animals similar to nematodes. Adults are free-living and reproduction takes place in freshwater environments, where preparasitic larvae undergo development. All species have a parasitic juvenil stage and infection may result in the host's death, insects being the most frequent host. Most of the life cycle occurs in freshwater environments, which are often contaminated by different pollutants. Based on the lack of information on the toxicity of herbicides to horsehair worms, the objective of this study is to evaluate the effect of different concentrations of glyphosate (technical grade and formulated product) on Chordodes nobilii (Gordiida, Nematomorpha). Bioassays were performed with embryos and larvae (preparasitic stages), and adults (postparasitic stage). Test organisms were exposed for a short period of time to concentrations ranging between 0.1 and 8 mga.e.l(-1) of glyphosate (technical and formulated). Although embryo development was not inhibited, there was a significant decrease in the infective capacity of larvae derived from eggs that had been exposed to >or= 0.1mg/l. Similar results were obtained for directly exposed larvae. No differences in toxicity were detected between the active ingredient and formulated product. Adult exposed for 96 h to 1.76 mgl(-1) formulated Gly shown a mortality of 50%. Results indicate that C. nobilii is affected at glyphosate concentrations lower than those expected to be found in freshwater environments and those specified in the legislation.     

Impaired glutathione redox status is associated with decreased survival in two organophosphate-poisoned marine bivalves

Biomonitoring organophosphate (OP) exposure in marine environments is generally achieved by the measurement of acetylcholinesterase activity in bivalves like mussels. However, there is evidence that indicates that oxidative stress may be implied in OP toxicity. The aim of this study was to evaluate the relationship between survival from the OP insecticide fenitrothion and glutathione levels in marine bivalves. Mussels (Mytilus galloprovincialis Lam.) and scallops (Flexopecten flexuosus Poli) were exposed, in a time to death test, to their LC85 of fenitrothion for 96 h. OP-poisoned mussels showed reduced (GSH) and oxidised (GSSG) glutathione depletion in the digestive gland, muscle and gills. Pectinid spats exposed to this insecticide presented GSH depletion in the digestive gland and mantle, and a reduction of the GSH/GSSG ratio in gills and mantle. Although survival curves were significantly different and mussels withstood twice as much fenitrothion as pectinid spats, muscular GSH/GSSG ratio was highly related to mortality in both species. We suggest that an impairment in the glutathione redox status could result in an induction of the cell death, either by apoptosis or necrosis, leading ultimately to the death of the organism. We conclude that whereas glutathione depletion can be used as a biomarker of exposure, the muscular GSH/GSSG ratio might be used as a biochemical marker of effect and individual susceptibility to mortality of marine bivalves exposed to fenitrothion or other pollutants that induce oxidative stress.     

S-metolachlor pulse exposure on the alga Scenedesmus vacuolatus: effects during exposure and the subsequent recovery

In streams and creeks, the aquatic flora is exposed to fluctuating concentrations of herbicides during and following their application. Peak concentrations of herbicides, like the chloroacetanilide S-metolachlor, are usually detected following rain events. In this study, we assessed the effect of S-metolachlor pulse exposure on the algae Scenedesmus vacuolatus. We measured the time-dependency of effects during exposure on algae population and identified the algae development stage most sensitive to S-metolachlor. Furthermore, we assessed the time-to-recovery of the algae following exposure. A 6h pulse exposure at 598mugl(-1) was sufficient to inhibit cell reproduction by 50%. However, the exposure period had to coincide with the cell development stage specifically inhibited by S-metolachlor, which is the end of the cell growth phase. In algae populations composed of cells at all development stages, we initially observed an increase in the size of some algal cells, ultimately leading to an inhibition of the growth rate. In these experimental conditions, effects were observed after 18h of exposure and greatly increased with time. The recovery of algae following exposure to strongly inhibiting S-metolachlor concentrations was delayed and only occurred after 29h. These findings suggest that peak exposure to S-metolachlor may affect the growth of sensitive alga in surface waters, considering that the effects extend beyond the period of exposure.     

Population dynamics in Daphnia Magna as modified by chronic tetradifon stress

Abstract

Two Daphnia magna offsprings (animals from the first and third brood) whose parentals (F₀‐generation) were exposed during 21 days to different tetradifon (4‐chlorophenyl 2,4,5‐trichlophenyl sulfone) concentrations, were transferred to a pesticide free medium during 21 days (recovery period). The algae Nannochloris oculata (5x10⁵ cells/mL) was used as food. In this recovery study, survival, growth and reproduction (mean total young per female, mean brood size, onset of reproduction and mean number broods per female) were assessed as individual parameters and the intrinsic rate of natural increase (r) as population parameter, for F₁ generation (1st and 3rd broods). Reproduction was still reduced in F₁ (1st and 3rd broods) generation daphnids from parentals (F₀) exposed to 0.18 mg/L tetradifon and higher concentrations even after 21 days in clean water. However, survival was not significantly different (p>0.05) in those F1 offsprings from parentals pre‐exposed to the acaricide. Growth was still reduced in F1 daphnids from parentals pre‐exposed to 0.10–0.44 mg/L tetradifon. The intrinsic rate of natural increase (r) was still affected in Fl generation daphnids, specially in those from the third brood. Therefore, Fl generation D. magna offsprings from a parental generation (F0) previously exposed to the acaricide tetradifon were not able to recuperate completely when a recovery period of 21 days was allowed.     

Reproductive impairment of sea urchins upon chronic exposure to cadmium. Part I: Effects on gamete quality

Successful reproduction is a determining factor for species survival. Pollution may impair reproductive success of adults through effects on gamete quality. Reproductive impairment of the sea urchin Anthocidaris crassispina upon chronic (4 weeks) exposure to 0.01 and 0.1 mg l-1 Cd2+ was investigated. Criteria used for assessing gamete quality included sperm motility, egg morphology, fertilization rate and dynamics of first cleavage. A dose-response relationship was found between Cd2+ levels and changes in sperm motility, and percentage fertilization. Sperm motility, measured by computer-assisted sperm analysis, indicated that percent motile sperm, velocities, and percent sperm with normal trajectory were significantly affected by chronic exposure to > or = 0.1 mg l-1 Cd2+. A decline in sperm motility was also accompanied by a decrease in fertilization success of sea urchin sperm. Width/height ratio of sea urchin eggs was not affected by cadmium, but larger egg sizes were found when sea urchins were exposed to 0.1 mg l-1 Cd2+. Male sea urchins exposed to Cd2+ produced poorer quality sperm, as indicated by a lower percent fertilization and lower cleavage rate, implying that male sea urchins were more sensitive than females to chronic Cd2+ exposure. Results of the present study provide an explanation of reproductive impairment in marine invertebrates upon chronic exposure to Cd2+.