Evaluation of multigenerational effects of 2-ethylhexyl 4-hydroxybenzoate in Japanese medaka.

The Japanese medaka (Oryzias latipes) extended one-generation reproduction test (MEOGRT) (Test Guideline 890.2200) is a Tier 2 test within the Endocrine Disruptor Screening Program of the US Environmental Protection Agency (US EPA). A modified MEOGRT was used to evaluate multigenerational effects of 2-ethylhexyl 4-hydroxybenzoate (2-EHHB) under flow-through conditions starting with adults (parent generation, F0) through a 3-week reproductive phase of the second generation (F2). Fish were exposed to one of five 2-EHHB test concentrations or a dechlorinated tap water control. Fecundity was affected at the lowest exposure (5.32 µg/L) and greater sensitivity occurred in the F1 and F2 generations. Percent fertility was also diminished from no effect level observed in the F0 generation to 101 and 48.8 µg/L in the F1 and F2 generations, respectively. Growth indices were decreased for F0 adult females and F1 subadults and adults at 48.8 µg/L 2-EHHB. Histopathologic examination of gonads, liver, kidney, and thyroid yielded possible delayed reproductive tract development in F1 subadult males, masculinization of the renal phenotype in F1 adult females (renal tubular eosinophilia) and reduced hepatic energy storage (liver glycogen vacuoles) in F1 (11.3 and 48.8 µg/L) and F2 (48.8 and 101 µg/L) males and females, respectively. Endocrine-related findings included a decrease in anal fin papillae in F2 adult males at 101 µg/L. Results of this study demonstrate effects on growth, development, and reproduction that may be mediated by endocrine (weak estrogenic) and nonendocrine mechanisms. Duration of the MEOGRT should not be routinely extended beyond the OCSPP 890 guideline study design.

Evaluation of the ecotoxicological effects of biogenic amines derived from cadaverous putrefaction on springtails Folsomia candida.

Necro-leachate, a liquid released during cadaveric decomposition, is considered the main culprit for impacts on cemetery environments. The biogenic amines cadaverine and putrescine make up part of the composition of necro-leachate and have a certain toxicity to different organisms. Springtails are among the most used bioindicators to assess the impacts of soil contaminants. As there are no data on the acute and chronic toxicity of springtails exposed to cadaverine and putrescine, the objective of this study was to evaluate the toxic potential of both amines, under the behavioral effect of avoidance and reproduction in the species Folsomia candida. Springtails were exposed to soils contaminated with different concentrations of cadaverine and putrescine, and different mixtures of these amines. To evaluate the avoidance and reproduction test, the individuals were exposed for periods of 48 h and 28 days, respectively. The results obtained in the avoidance test showed that springtails exhibited a preference for the treated soil in both isolated and mixed treatments. The chronic evaluation assays showed that the reproduction was affected, particularly in the treatments with combined amines, resulting in a reduction in the total number of juveniles. From the results, it is possible to infer that the methods applied in this research have provided information that will contribute to a better understanding of the toxicity of putrefactive biogenic amines, since there exist few ecotoxicological studies carried out with these amines, and especially with those from cemetery environments.

The value of avian gross pathology in identifying endocrine disrupting properties.

The European Chemical Agency and European Food Safety Authority recommend that gross pathology findings, from avian reproduction studies, be used to support assessment of potential endocrine disrupting properties of active pesticidal and biocidal substances. In open literature, little information is available on the utility of gross pathology data for informing endocrine evaluations. Here the gross pathology data from historical control groups of 51 northern bobwhite and 51 mallard reproduction tests is analyzed to evaluate the utility of such information. Incidence of gross morphology findings in untreated birds may aid the interpretation of some gross abnormalities, potentially indicative of an endocrine interaction (e.g. reproductive condition). Statistical analysis of the historical control data indicates that gross pathology is not likely to be useful for detecting endocrine effects as abnormalities with relatively high increases in prevalence (more than 20-30%, depending on prevalence in controls) are reliably interpreted as a treatment response. Gross pathology changes are only indicative and not diagnostic of endocrine interactions making it difficult to distinguish gross pathology abnormalities, due to endocrine-mediated effects, from systemic toxicity. This work demonstrates the utility of using historical control analyses to establish the value and properties of selected endpoints for regulatory applications.

Assessment of the reproduction of six collembolan species in tropical soils naturally rich in potentially toxic elements.

Laboratory ecotoxicological tests are important tools for the management of environmental changes derived from anthropogenic activities. Folsomia candida is usually the model species used in some procedures. However, this species may not be sufficiently representative of the sensitivity of the other collembolan species. This study aimed to evaluate (i) the effects of soils naturally rich in potentially toxic elements (PTE) and soil characteristics on the reproduction and survival of different collembolan species, (ii) whether the habitat function of these soils is compromised, and (iii) to what extent F. candida is representative of the other collembolan species. For this, reproduction tests with six collembolan species were conducted in 14 different samples of soils. In general, collembolan reproduction was not completely inhibited in none of the natural tested soils. Even soils with high pollution load index values did not negatively affect collembolan reproduction for most of the species. In contrast, the lowest collembolan reproduction rates were found in a visually dense soil (lowest volume/weight ratio), highlighting that soil attributes other than total PTE concentration also interfere in the reproduction of collembolans. Our results support the idea that the F. candida species might not be representative of other collembolan species and that laboratory tests to assess soil contaminations should be conducted using diverse collembolan species.

From the importance of organic carbon in reproduction tests with Eisenia fetida.

The standard Eisenia fetida chronic toxicity reproduction test is normalized and needs up to 8 weeks of experimentation. Many matrixes have shown toxic effects on the earthworms' survey, growth and reproduction. But the food factor may affect the organisms' responses to contaminated media in such tests. We try to evaluate the effect, or not, of the organic carbon content present in the environment, as spreadable organic wastes added in soil, on the reproduction test of Eisenia fetida. We confirm the influence of the organic matter content on earthworm reproduction. This "food" factor must be considered when studying environmental samples rich in organic matter, since that could mask potentially harmful effects. To minimize this potential "food" effect during reproduction tests, we recommend an inverse gradient of organic matter contribution by the "horse dung" food applied in all conditions according to the dose of agricultural amendments tested.

Variation in chronic nickel toxicity to Daphnia magna among Japanese river waters and performance evaluation of bioavailability models in predicting the toxicity.

Nickel (Ni) ecotoxicity is dictated by water chemistry characteristics such as pH, water hardness, and amount of dissolved organic carbon. Bioavailability models have been developed to predict Ni toxicity and validated for European, Australian, and US natural waters. In this study, chronic toxicity tests in Ni-spiked Japanese river waters were conducted on a strain of Daphnia magna to test whether the chronic toxicity differs among Japanese natural waters with different water chemistries. Based on the results of chronic Ni toxicity tests, we assessed the performance of existing D. magna bioavailability models, which were developed in artificial waters (Model 1) and calibrated in European natural waters (Model 2), in terms of the accuracy and the bias of model predictions. Furthermore, we also calibrated the two models by using toxicity test results to develop a bioavailability model for Ni chronic toxicity to the strain of D. magna in Japanese river waters. The 10%, 20%, and 50% effect concentrations (EC10, EC20, and EC50) of dissolved Ni on reproduction of the D. magna strain were within ranges from 8.1 to 44.9 µg/L, 9.0 to 57.1 µg/L, and 10.9 to 86.1 µg/L, respectively. Results indicate that differences in water chemistry among Japanese river waters influenced chronic Ni toxicity to the model organism. Model 1predicted 43% of the observed EC10, EC20, and EC50 values within a factor of 2 and 100%, 100%, and 43% within a factor of 3, respectively. Model 2 predicted 14%, 14%, and 29% of the observed EC10, EC20, and EC50 values within a factor of 2 and 43% within a factor of 3. The values of model bias based on the geometric mean of ratios of EC10, EC20 and EC50 values predicted by each of the two models and observed EC10, EC20, and EC50 values were 0.71, 0.65, and 0.62 for Model 1 and 0.27, 0.26, and 0.29 for Model 2, respectively. After calibrating two models using the results of toxicity tests, refined Model 1 predicted 71%, 57%, and 57% of observed EC10, EC20, and EC50 values within a factor of 2 and 100%, 86%, and 100% within a factor of 3; refined Model 2 predicted 71% of observed EC10, EC20, and EC50 values within a factor 2 and 100%, 86%, and 86% within a factor of 3, respectively. Our results indicate that calibrating the Ni bioavailability models in Japanese natural waters increased their predictive capacity by a factor of up to approximately five.

Ecological risk assessment of an antifouling biocide triphenyl (octadecylamine) boron in the Seto Inland Sea, Japan.

In this study, we derived the predicted no-effect concentrations (PNEC) for triphenyl (octadecylamine) boron (TPB-18) and investigated the occurrence of triphenylboranes (TPBs), including TPB-18, for ecological risk assessment in the Seto Inland Sea, Japan. We tested algal growth inhibition, crustacean immobilization, and reproductive toxicity and performed toxicity tests in fish to assess acute and chronic toxicity and generate the PNEC for TPB-18. The minimum toxicity value was 0.30 µg/L, as determined by the 72-h no-observed-effect concentration (NOEC) for the alga Chaetoceros gracilis. The 5th-percentile of hazardous concentration (HC5), derived from NOECs using the species sensitivity distributions approach, was 0.059 µg/L, which indicated the PNEC of 0.0059 µg/L. In comparison, the highest concentration in seawater sampled from the Seto Inland Sea was 0.00034 µg/L, suggesting that the ecological risks posed by TPB-18 are currently low.

The ecological risk assessment of soil contamination with Ti and Fe at military sites in Ukraine: avoidance and reproduction tests with Folsomia candida.

The aim of this research was to assess the effect of soil contamination with titanium (Ti) and iron (Fe) at military sites in Ukraine using the avoidance and reproduction tests with Folsomia candida (springtail). The soil used for the tests was sampled in 2017 from Dolyna, Ivano-Frankivsk region, Ukraine from two plots, namely a contaminated and a control site. The sample site is a former military site previously used for tank training. At the control site mainly the concentrations of Ti and Fe were exceeded. The control soil was free from contamination. The avoidance test and reproduction test were conducted with the use of springtail species F. candida. The following nine levels of contamination with heavy metals were established: 1%, 1.5%, 5%, 10%, 15%, 25%, 50%, 75% and 100%. The duration of the avoidance test was 7 days, and that of the reproduction test was 28 days. Overall, the results show that the avoidance and reproduction tests with collembolans have the potential to be used as screening tools in an ecological risk assessment of heavy metals. In the avoidance test, the concentrations from 1.5 to 100% significantly decreased the number of F. candida in the contamination site in comparison to the control site. At the same time, avoidance was not observed in the first concentration (1%). According to the reproduction test, the negative effect on the number of F. candida juveniles was observed beginning at the 10% dose. The half maximal effective concentration (EC50) for the avoidance test was 50.12%, while that for the reproduction test was 22.39%. The contamination with heavy metals at the military areas indicated the short- and long-term toxicity risk on the springtail F. candida.

Sublethal and chronic effects of reclaimed water on aquatic organisms. Looking for relationships between physico-chemical characterisation and toxic effects.

The use of reclaimed water for irrigation and aquaculture purposes is generally considered a reliable alternative for sustainable water management in regions with water scarcity. Many organic compounds, generally called compounds of emerging concern (CECs), have been detected in reclaimed water, which implies continuous exposure for aquatic organisms. To date no quality criteria have been proposed for this group of compounds. This work aims to assess the acute, sublethal and chronic effects of reclaimed water using two representative organisms of the aquatic compartment; the green alga Chlorella vulgaris and the microcrustacean Daphnia magna. The study comprises the 72 h-algal growth inhibition test, the D. magna feeding bioassay and the D. magna reproduction test. The results highlighted, for the selected characterised compounds, no differences in the concentrations between the different tertiary WWTP treatments, except for the particular case of carbamazepine. Considering seasonality, no differences were observed between the two different sample collection campaigns. The sublethal and chronic effects observed for these samples could not be explained by the lower concentrations found in the chemical characterisation. However, in the majority of cases, dilution of raw reclaimed water reduced the toxic effects of these samples. Several interactions among compounds can affect the mixture's toxicity. Canonical correlation analyses (CCA) were included to explore the potential relationships between the physico-chemical characterisation of reclaimed water and effects on aquatic organisms. The results corroborated the toxic effect of some pharmaceuticals, in particular beta-blockers and antibiotics, on the growth and yield of green algae, as well as inhibition of daphnia reproduction. Thus the CCA methods could help to elucidate the potential relationships between the physico-chemical characterisation and toxic effects by considering all the potential interactions.

Summary of the development the US Environmental Protection Agency's Medaka Extended One Generation Reproduction Test (MEOGRT) using data from 9 multigenerational medaka tests.

In response to various legislative mandates, the US Environmental Protection Agency (USEPA) formed its Endocrine Disruptor Screening Program (EDSP), which in turn, formed the basis of a tiered testing strategy to determine the potential of pesticides, commercial chemicals, and environmental contaminants to disrupt the endocrine system. The first tier of tests is intended to detect the potential for endocrine disruption mediated through estrogen, androgen, or thyroid pathways, whereas the second tier is intended to further characterize the effects on these pathways and to establish a dose-response relationship for adverse effects. One of these tier 2 tests, the Medaka Extended One Generation Reproduction Test (MEOGRT), was developed by the USEPA for the EDSP and, in collaboration with the Japanese Ministry of the Environment, for the Guidelines for the Testing of Chemicals of the Organisation for Economic Co-operation and Development (OECD). The MEOGRT protocol was iteratively modified based on knowledge gained after the successful completion of 9 tests with variations in test protocols. The present study describes both the final MEOGRT protocol that has been published by the USEPA and the OECD, and the iterations that provided valuable insights into nuances of the protocol. The various tests include exposure to 17ß-estradiol, 4-t-octylphenol, o,p'- dichlorodiphenyltrichloroethane, 4-chloro-3-methylphenol, tamoxifen, 17ß-trenbolone, vinclozolin, and prochloraz. Environ Toxicol Chem 2017;36:3387-3403. Published 2017 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

How does reverse shoulder replacement change proprioception in patients with cuff tear arthropathy? A prospective optical 3D motion analysis study.

BACKGROUND: Reverse total shoulder arthroplasty (RSA) can improve function in cuff tear arthropathy (CTA) shoulders, but limited data are available on the effect of RSA on proprioception. The purpose of this study was the evaluation of the proprioception after RSA. METHOD: This study included fifteen consecutive patients (n = 15) who received an RSA. We measured an active angle reproduction (AAR) by a novel 3D motion analysis using the Heidelberg upper extremity model (HUX). RESULTS: The overall proprioception didn't change significantly from 6.2° preoperatively to 6.2° postoperatively. CONCLUSION: We conclude that, the overall proprioception remained the same or deteriorated for different movements.

Medaka extended one-generation reproduction test evaluating 4-nonylphenol.

The medaka extended one-generation test (MEOGRT) was developed as a multigenerational toxicity test for chemicals, particularly endocrine-disrupting chemicals. Briefly, 3 generations of Japanese medaka (Oryzias latipes) are exposed to a chemical over a 20-wk period: 3 wk in the parental generation (F0), 15 wk in the first generation (F1), and 2 wk in the second generation (F2). The present study reports the first MEOGRT results concerning branched isomer mixtures of 4-nonylphenol (NP). Adult F0 medaka exposed to NP at 5 actual concentrations (1.27, 2.95, 9.81, 27.8, 89.4 µg/L) were unaffected in terms of reproduction, although vitellogenin in the male liver was increased dose-dependently at concentration of 2.95 µg/L and higher. In F1, in contrast, total egg (fecundity), fertile egg, and fertility decreased as NP increased; lowest-observed-effect concentrations (LOECs) for total egg, fertile egg, and fertility were 1.27, 1.27, 27.8 µg/L, respectively. In F1, but not in F0, secondary sex characteristics (i.e., anal fin papillae in males) were suppressed at 27.8 µg/L NP. Vitellogenin induction in adult male fish was slightly weaker in F1 than it was in F0, however. Gonadal sex abnormality and sex reversal occurred at 27.8 and 89.4 µg/L NP in F1 subadults. At 89.4 µg/L NP, all genotypic F1 males in breeding pairs had female phenotype, and some even demonstrated spawning. Concentrations of NP lower than 89.4 µg/L did not affect F2 survival or hatching. The highest detected NP level in environmental freshwater in Japan was approximately a half of the LOEC (1.27 µg/L for F1 fecundity); in other countries, however, environmental concentrations above the LOEC are reported, suggesting that NP may be affecting fish populations. Environ Toxicol Chem 2017;36:3254-3266. © 2017 SETAC.

Two-generational effects of contaminants in Daphnia magna: Effects of offspring quality.

The authors set up a protocol to perform a 2-generational ring test using the existing guidelines for the Daphnia magna reproduction test. It is well known in ecology that size and quality of offspring vary across the first clutches in D. magna and that certain chemicals affect offspring quality. Therefore, the origin of the second generation is an important factor to consider. Two-generational effects across first, second, and third clutches were evaluated using 4-nonylphenol; those across first and third clutches were evaluated using tributyltin, and those across the third clutch were evaluated using piperonyl butoxide. The compound showing the greatest aggravation of toxic effects between the parental and second generations was piperonyl butoxide, followed by 4-nonylphenol, whereas intergenerational effects of tributyltin varied across experiments. The studied chemicals affected the quantity and quality of the offspring produced by exposed females of the parental generation, those effects being greater in third-clutch neonates. Therefore, when third-clutch offspring were further exposed, they turned out to be more sensitive than the parental generation. The results are in line with those obtained in multigenerational studies using mammalian tests, which showed that, in many cases, effects on the second generation can be predicted by evaluating the quality of the offspring produced. Environ Toxicol Chem 2016;35:1470-1477. © 2015 SETAC.

Comparison of earthworm responses to petroleum hydrocarbon exposure in aged field contaminated soil using traditional ecotoxicity endpoints and 1H NMR-based metabolomics.

(1)H NMR metabolomics and conventional ecotoxicity endpoints were used to examine the response of earthworms exposed to petroleum hydrocarbons (PHCs) in soil samples collected from a site that was contaminated with crude oil from a pipeline failure in the mid-1990s. The conventional ecotoxicity tests showed that the soils were not acutely toxic to earthworms (average survival ≥ 90%), but some soil samples impaired reproduction endpoints by >50% compared to the field control soil. Additionally, metabolomics revealed significant relationships between earthworm metabolic profiles (collected after 2 or 14 days of exposure) and soil properties including soil PHC concentration. Further comparisons by partial least squares regression revealed a significant relationship between the earthworm metabolomic data (collected after only 2 or 14 days) and the reproduction endpoints (measured after 63 days). Therefore, metabolomic responses measured after short exposure periods may be predictive of chronic, ecologically relevant toxicity endpoints for earthworms exposed to soil contaminants.