Protective role of humic acids against picloram-induced genomic instability and DNA methylation in <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>

Picloram (4-amino-3,5,6-trichloropicolinic acid) is a liquid auxinic herbicide used to control broad-leaved weeds. Picloram is representing a possible hazard to ecosystems and human health. Therefore, in this study, DNA methylation changes and DNA damage levels in Phaseolus vulgaris exposed to picloram, as well as whether humic acid (HA) has preventive effects on these changes were investigated. Random amplified polymorphic DNA (RAPD) techniques were used for identification of DNA damage and coupled restriction enzyme digestion-random amplification (CRED-RA) techniques were used to detect the changed pattern of DNA methylation. According to the obtained results, picloram (5, 10, 20, and 40 mg/l) caused DNA damage profile changes (RAPDs) increasing, DNA hypomethylation and genomic template stability (GTS) decreasing. On the other hand, different concentrations of applied HA (2, 4, 6, 8, and 10%) reduced hazardous effects of picloram. The results of the experiment have explicitly indicated that HAs could be an alternative for reducing genetic damage in plants. In addition to the alleviate effects of humic acid on genetic damage, its epigenetic effect is hypomethylation.     

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.     

Effects of dimethoate on rotifer Brachionus calyciflorus using multigeneration toxicity tests

As important members of the zooplankton community and sources of food for fish, rotifers are used extensively in ecotoxicological research to assess the health of the environment and safety of compounds. However, most rotifer toxicity tests are only conducted using rotifer neonates derived from unexposed mothers, thus ignoring the potential transfer of contaminants from mother to offspring. To understand better the mother to offspring exposure, a multigenerational study was conducted using three successive generations (F₀, F₁ and F₂) of the common freshwater rotifer Brachionus calyciflorus to investigate the toxic effects of the widely used organophosphate pesticide, dimethoate (O, O-dimethyl S-methylcarbamoylmethyl phosphorodithioate). When the F₀ generation was exposed to five pesticide concentrations, the population growth rate (r) displayed symptoms of hormesis, characterized by the conversion of low-concentration stimulation to high-concentration inhibition. Despite this observation, the exposure to any given concentration of dimethoate reduced the population growth rates of the F₁ and F₂ generation rotifers. Significant differences existed between the F₀, F₁ and F₂ rotifers for the population growth rate under dimethoate stress: F₂ individuals were more sensitive than F₁, whereas the F₁ individuals were more sensitive than F₀. The results indicated that the parental exposure to a given toxic stress could result in increased sensitivity and decreased fitness in the offspring. This study illustrates the utility of multigenerational toxicity tests, which may better reflect and more accurately predict the effects of long-term pesticide exposure to aquatic organisms at the population level.     

Screening differentially expressed genes in an amphipod (Hyalella azteca) exposed to fungicide vinclozolin by suppression subtractive hybridization

Vinclozolin, a dicarboximide fungicide, is an endocrine disrupting chemical that competes with an androgenic endocrine disruptor compound. Most research has focused on the epigenetic effect of vinclozolin in humans. In terms of ecotoxicology, understanding the effect of vinclozolin on non-target organisms is important. The expression profile of a comprehensive set of genes in the amphipod Hyalella azteca exposed to vinclozolin was examined. The expressed sequence tags in low-dose vinclozolin-treated and -untreated amphipods were isolated and identified by suppression subtractive hybridization. DNA dot blotting was used to confirm the results and establish a subtracted cDNA library for comparing all differentially expressed sequences with and without vinclozolin treatment. In total, 494 differentially expressed genes, including hemocyanin, heatshock protein, cytochrome, cytochrome oxidase and NADH dehydrogenase were detected. Hemocyanin was the most abundant gene. DNA dot blotting revealed 55 genes with significant differential expression. These genes included larval serum protein 1 alpha, E3 ubiquitin-protein ligase, mitochondrial cytochrome c oxidase, mitochondrial protein, proteasome inhibitor, hemocyanin, zinc-finger–containing protein, mitochondrial NADH-ubiquinone oxidoreductase and epididymal sperm-binding protein. Vinclozolin appears to upregulate stress-related genes and hemocyanin, related to immunity. Moreover, vinclozolin downregulated NADH dehydrogenase, related to respiration. Thus, even a non-lethal concentration of vinclozolin still has an effect at the genetic level in H. azteca and presents a potential risk, especially as it would affect non-target organism hormone metabolism.     

Effects of multigenerational exposures of D. magna to environmentally relevant concentrations of pentachlorophenol

The re-emergence of schistosomiasis has given rise to ubiquitous concentrations of the primary control agent pentachlorophenol (PCP) in the environment, especially in the surface waters of China. In this study, the effects of environmentally relevant concentrations of PCP, namely, 0.0002, 0.002, 0.02, 0.2, and 2 μmol/L on survival, age at first reproduction, fecundity, length of mothers, and number of molts of Daphnia magna were studied over three generations. The survival of D. magna exposed to 2 μmol/L was significantly affected in the three generations. Toxic effects were enhanced in later generations. Age at first reproduction of F ₁ and F ₂ D. magna was significantly slower than that of the controls. The total number of offspring per female exposed to concentrations of 0.002 μmol/L or greater was less (23.5 to 67.6, 9.4 to 73.7, and 3.6 to 83.7 %) than that of the controls in the F ₀, F ₁, and F ₂ generations, respectively. The body length of mothers significantly decreased (4.7 to 6.8, 9.6 to 15.1, and 13.3 to 23.2 %) after exposure to 0.002 μmol/L or greater than those of unexposed individuals in the F ₀, F ₁, and F ₂ generations, respectively. Dose–response relationships between concentrations of PCP and length and number of molts of D. magna were observed in the F ₀ to F ₂ generations. PCP concentrations on the surface waters of China caused adverse effects to D. magna, which increased over successive generations. Significant effects were observed in the third generation. The multigenerational studies were more sensitive than the single-generation experiments. Thus, multigenerational exposure may be more predictive of chronic exposure under field conditions.     

Arsenic concentration in topsoil of central Chile is associated with aberrant methylation of P53 gene in human blood cells: a cross-sectional study

Gene expression can be modified in people who are chronically exposed to high concentrations of heavy metals. The soil surrounding the Ventanas Industrial Complex, located on the coastal zone of Puchuncaví and Quintero townships (Chile), contain heavy metal concentrations (As, Cu, Pb, Zn, among others) that far exceed international standards. The aim of this study was to determine the potential association of the heavy metals in soils, especially arsenic, with the status of methylation of four tumor suppressor genes in permanent residents in those townships. To study the methylation status in genes p53, p16, APC, and RASSF1A, we took blood samples from adults living in areas near the industrial complex for at least 5 years and compared it to blood samples from adults living in areas with normal heavy metal concentrations of soils. Results indicated that inhabitants of an area with high levels of heavy metals in soil have a significantly higher proportion of methylation in the promoter region of the p53 tumor suppressor gene compared with control areas (p-value: 0.0035). This is the first study to consider associations between heavy metal exposure in humans and aberrant DNA methylation in Chile. Our results suggest more research to support consistent decision-making on processes of environmental remediation or prevention of exposure.

     

Effects of aqueous stable fullerene nanocrystals (nC60) on Daphnia magna: Evaluation of sub-lethal reproductive responses and accumulation

Concerns exist regarding the inadvertent release of engineered nanomaterials into natural systems, and the possible negative ecosystem response that may occur. Understanding sub-lethal effects may be particularly important to determining ecosystem responses as current levels of nanomaterial release are low compared to levels projected for the future. In this work, the sub-lethal effects and bioaccumulation of water stable, nanocrystalline fullerenes as C₆₀, (termed nC₆₀) were studied in Daphnia magna, a globally distributed, parthenogenetic zooplankton. Sub-lethal concentrations were first determined for both mature mother (LD₅₀ = 0.4 mg L⁻¹) and neonate (gestating) daphnids (0.2 mg L⁻¹) in standard 48 h exposure tests. Subsequent experiments focused on the accumulation and effects (at temperatures of 18–28 °C) of nC₆₀, during the D. magna reproductive cycle. The results demonstrate that upon sub-lethal exposure, the mortality rates of gestating daphnids increased with time and developmental stage. The maturation of daughter daphnids was negatively impacted. The mother daphnids were unable to reproduce again after exposure during pregnancy, and differential bioaccumulation occurred as a function of lipid content in the daphnia with the highest accumulation level of 7000 mg kg⁻¹ wet weight. Taken together, these results not only describe the accumulation and sub-lethal effects of nC₆₀ on exposed daphnia, but also highlight the importance of sub-lethal exposure scenarios, which are critical to fully understanding the potential impact of fullerenes and other engineered nanoscale materials on natural systems.     

As-resistance in laboratory-reared F1, F2 and F3 generation offspring of the earthworm Lumbricus rubellus inhabiting an As-contaminated mine soil

Previous studies provided no unequivocal evidence demonstrating that field populations of Lumbricus rubellus Hoffmeister (1843), exhibit genetically inherited resistance to As-toxicity. In this study F1, F2 and F3 generation offspring derived from adults inhabiting As-contaminated field soil were resistant when exposed to 2000 mg kg⁻¹ sodium arsenate. The offspring of uncontaminated adults were not As-resistant. Cocoon viability was 80% for F1 and 82% for F2 offspring from As-contaminated adults and 59% in the F1 control population. High energy synchrotron analysis was used to determine whether ligand complexation of As differed in samples of: resistant mine-site adults, the resistant F1 and F2 offspring of the mine-site earthworms exposed to the LC₂₅ sodium arsenate (700 mg kg⁻¹) of the F1 parental generation; and adult L. rubellus from an uncontaminated site exposed to LC₂₅ concentrations of sodium arsenate (50 mg kg⁻¹). XANES and EXAFS indicated that As was present as a sulfur-coordinated species.     

Chronic depression-like phenotype in male offspring mice following perinatal exposure to naturally contaminated eels with a mixture of organic and inorganic pollutants

Previously, we demonstrated that maternal exposure to high, intermediate, or lowly contaminated European eels with a mixture of chemicals, during pregnancy and lactation, resulted in adult despair-like behavior, selectively in male offspring mice. Here, we investigate if depression-like behavior in offspring males was transient or permanent by monitoring immobility behavior, a measure of behavioral despair, at three distinct stages of life, including young adult (post-natal day (PND) 55), mature adult (PND 200) and middle (PNDs 335–336) age, in the forced swimming (FST) and the tail suspension (TST) tests. Oxidative stress markers including malondialdehyde (MDA) levels and superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities were evaluated in the hippocampus, prefrontal cortex, and cerebellum of middle-aged animals. Findings showed a significant enhancement of immobility behavior in the TST performed at young adult age (all p < 0.05) in the FST carried out at mature adult age (all p < 0.001) and in both behavioral tests realized at middle age (all p < 0.05, except one p = 0.06) in mice perinatally exposed to eels compared with non-exposed controls. Antioxidant-related enzyme activities, including SOD and CAT, were only elevated in the hippocampus of middle-aged males perinatally exposed to the two more polluted eels (all p < 0.05). Further, lipid peroxidation, assessed by MDA levels, was not found to be differentially regulated in the selected areas of middle-aged brains of exposed mice (all p > 0.05). Collectively, this suggested limited oxidative metabolism disturbances in middle-aged brains exposed to eels. In summary, our results highlighted that offspring males perinatally exposed to naturally contaminated reared and river eels with persistent organic pollutants (POPs) and heavy metals displayed chronic depression-like phenotype. As extrapolation of data to humans should be done with precaution, retrospective and prospective epidemiological studies are needed to clarify this potential relationship, stressed in our animal model, between maternal polluted fish consumption and chronically low mood in offspring.

     

Juvenile hormone agonists affect the occurrence of male Daphnia

The water flea Daphnia magna reproduces primarily by cyclic parthenogenesis. Environmental stimuli that signal a change to adverse conditions induce the organisms to switch from parthenogenesis to gamogenetic reproduction. During the gamogenetic period, they produce male daphnids and dormant resting eggs, which can survive prolonged periods of environmental adversity. However, little is known about the mechanisms associated with the switch from parthenogenesis to gamogenetic reproduction. We investigated the effects of several juvenoids on sex determination in Daphnia. Females less than 24 h old were exposed to various concentrations of the test substance and were observed for 21 days. It was found that they can trigger the appearance of male daphnids: the percentage of males in the population increases to a level greater than what occurs under ordinary environmental conditions. We found that methylfarnesoate, juvenile hormone III, methoprene, and the phenoxyphenoxy derivatives pyriproxyfen and fenoxycarb (both insecticides) reduced the production of offspring and produced sex ratios dominated by male daphnids. Pyriproxyfen and fenoxycarb showed striking effects at low concentrations. Exposure to either of these chemicals at a concentration of 330 ngl(-1) caused adult females to produce almost all male neonates. Methylfarnesoate, juvenile hormone III, and methoprene showed an effect in inducing male production at higher concentrations (3.7 x 10(3), 3.3 x 10(5), and 1.3 x 10(5) ngl(-1), respectively). Our findings suggest that juvenile hormone agonists, including some insecticides, affect the chemical signaling responsible for inducing the production of male offspring.     

A two-generational reproductive toxicity study of zinc in rats

A two-generation reproductive toxicity study of zinc chloride (ZnCl₂) was conducted in rats. F_o male and female rats were administered 0.00 (control), 7.50 (low), 15.00 (mid) and 30.00 (high) mg/kg/day of ZnCl₂. Selected F₁ male and female rats were exposed to the same doses received by their parents (F_o). Exposure of F₀ parental rats to ZnCl₂ showed significant reduction in fertility, viability (days 0 and 4), and the body weight of F₁ pups from the high-dose group but caused no effects on litter size, weaning index, and sex ratio. Similarly, the continued exposure of F₁ parental rats to ZnCl₂ also reduced fertility, liter size, viability (day 0), and the body weight of F₂ pups within the high-dose group but caused no effects on weaning index and sex ratio. Exposure of ZnCl₂ to F₀ and F₁ parental males resulted in a significant reduction in their body weights, and the F₀ and F₁ parental females did not show any significant difference in their body weights compared to their control groups. However, the postpartum dam weights of both F₀ and F₁ female rats were significantly reduced compared to their controls. Exposure of ZnCl₂ to F_o and F₁ generation parental rats did not produce any significant change of their clinical signs as well as their clinical pathology parameters, except the alkaline phosphotase (ALK) level, which showed an upward trend in both sexes of both generations. Exposure of ZnCl₂ to F₀ rats resulted in a reduction of brain, liver, kidney, spleen and seminal vesicles weights of males and in the spleen and uterus of females. Similarly, exposure of F₁ rats to ZnCl₂ also resulted in reduction of brain, liver, kidney, adrenal, spleen, prostate and seminal vesicles weights of males and in spleen and uterus of females. ZnCl₂ exposure resulted in grossly observed gastro-intestianla (GI) tract, lymphoreticular/hematopoietic, and reproductive tract lesions in parental rats in both generations. Reduced body fat was also recorded in F₁ parental rats.     

Toxicity of silver nanoparticles to rainbow trout: A toxicogenomic approach

Silver (Ag) nanoparticles are used as antimicrobial adjuvant in various products such as clothes and medical devices where the release of nano-Ag could contaminate the environment and harm wildlife. The purpose of this study was to examine the sublethal effects of nano-Ag and dissolved Ag on Oncorhynchus mykiss rainbow trout. Hepatic Ag contents and changes in gene expression were monitored to provide insights on bioavailability and mode of action of both forms of silver. Fish were exposed to increasing concentrations (0.06, 0.6 and 6 μg L⁻¹) of nano-Ag (20 nm) and silver nitrate (AgNO₃) for 96 h at 15 °C. A gene expression analysis was performed in the liver using a DNA microarray of 207 stress-related genes followed by a quantitative polymerase chain reaction on a selection of genes for validation. The biochemical markers consisted of the determination of labile zinc, metallothioneins, DNA strand breaks, lipid peroxidation (LPO) and vitellogenin-like proteins. The analysis of total Ag in the aquarium water revealed that nano-Ag was mostly aggregated, with 1% of the total Ag being dissolved. Nevertheless, hepatic Ag content was significantly increased in exposed fish. Indeed, dissolved Ag was significantly more bioavailable than nano-Ag only at the highest concentration with 38 ± 10 and 11 ± 3 ng Ag mg⁻¹ proteins for dissolved and nano-Ag respectively. Exposure to both forms of Ag led to significant changes in gene expression for 13% of tested gene targets. About 12% of genes responded specifically to nano-Ag, while 10% of total gene targets responded specifically to dissolved Ag. The levels of vitellogenin-like proteins and DNA strand breaks were significantly reduced by both forms of Ag, but DNA break levels were lower with nano-Ag and could not be explained by the presence of ionic Ag. Labile zinc and the oxidized fraction of metallothioneins were increased by both forms of Ag, but LPO was significantly induced by nano-Ag only. A discriminant function analysis revealed that the responses obtained by biochemical markers and a selection of ten target genes were able to discriminate completely (100%) the effects of both forms of Ag. Exposure to nano-Ag involved genes in inflammation and dissolved Ag involved oxidative stress and protein stability. Hence, the toxicity of Ag will differ depending on the presence of Ag nanoparticles and aggregates.     

Assessment of the effects of the carbamazepine on the endogenous endocrine system of Daphnia magna

In the present study, the endocrine activity of the antiepileptic pharmaceutical carbamazepine (CBZ) in the crustacean Daphnia magna was assessed. To assess the hormonal activity of the drug, we exposed maternal daphnids and embryos to environmental relevant concentrations of CBZ (ranging from 10 to 200 μg/L) and to mixtures of CBZ with fenoxycarb (FEN; 1 μg/L). Chronic exposure to CBZ significantly decreased the reproductive output and the number of molts of D. magna at 200 μg/L. This compound induced the production of male offspring (12?±?1.7 %), in a non-concentration-dependent manner, acting as a weak juvenile hormone analog. Results showed that this substance, at tested concentrations, did not antagonize the juvenoid action of FEN. Further, CBZ has shown to be toxic to daphnid embryos through maternal exposure interfering with their normal gastrulation and organogenesis stages but not producing direct embryo toxicity. These findings suggest that CBZ could act as an endocrine disruptor in D. magna as it decreases the reproductive output, interferes with sex determination, and causes development abnormality in offspring. Therefore, CBZ could directly affect the population sustainability.

     

Growth and Foliar Nitrogen Status of Four Plant Species Exposed to Atmospheric Ammonia

A chamber study was conducted to evaluate the growth response and leaf nitrogen (N) status of four plant species exposed to continuous ammonia (NH₃) for 12 weeks (wk). This was intended to evaluate appropriate plant species that could be used to trap discharged NH₃ from the exhaust fans in poultry feeding operations before moving off-site. Two hundred and forty bare-root plants of four species (Juniperus virginiana (red cedar), Gleditsia triacanthos var. inermis (thornless honey locust), Populus sp. (hybrid poplar), and Phalaris arundinacea (reed canary grass) were transplanted into 4- or 8-L polyethylene pots and grown in four environmentally controlled chambers. Plants placed in two of the four chambers received continuous exposure to anhydrous NH₃ at 4 to 5 ppm while plants in another two chambers received no NH₃. In each of the four chambers, 2 to 4 plants per species received no fertilizer while the rest of the plants were fertilized with a 100 ppm solution containing 21% N, 7% phosphorus, and 7% potassium. The results showed that honey locust was the fastest-growing species. The superior growth of honey locust among all species was also supported by its total biomass, root, and root dry matter (DM) weights. For all species there was a trend for plants exposed to NH₃ to have greater leaf DM than their non-exposed counterparts at 6 (43.0 vs. 30.8%; P = 0.09) and 12 wk (47.9 vs. 36.6%; P = 0.07), and significantly greater (P ≤ 0.05) leaf N content at 6 (6.44 vs. 3.67%) and 12 wk (7.05 vs. 3.51%) when exposed to NH₃. Numerically greater leaf DM due to NH₃ exposure was also consistently measured in poplar at both sampling periods. Hybrid poplar, as well as honey locust and reed canary grass, deposited 1.5 to 2-fold greater N in their leaves than red cedar tissues as a result of NH₃ exposure compared to non-exposed plants. Regardless of the effect of NH₃ on foliar color and damage score of the plants, the increase of foliar N content (g 100 g^?1 of fresh foliage weight) after NH₃ exposure at 6 and 12 wk was 0.45 and 0.87 for grass,1.25 and 1.34 for locust, and 2.67 and 6.09 for poplar. However, only honey locust likely benefited from ambient NH₃ as indicated by its consistent leaf color quality and lower damage score, compared with other species that were adversely affected by atmospheric NH₃.     

Short term recovery of periphyton photosynthesis after pulse exposition to the photosystem II inhibitors atrazine and isoproturon

Aquatic organisms are exposed to fluctuating concentrations of herbicides which contaminate rivers following their use for agricultural or domestic purposes. The development of sensitive bioanalytical tests enabling us not only to detect the effects of those pollutants but to take into account this pattern of exposure should improve the ecological relevance of river toxicity assessment. In this respect, the use of chlorophyll fluorescence measurements is a convenient way to probe the effect of photosystem II (PSII) inhibitors on primary producers. This study was devoted to validate the combined use of two fluorescence parameters, the effective and the optimal quantum yields of PSII photochemistry (Φ_PSII and F_v/F_m), as reliable biomarkers of initial isoproturon (IPU) or atrazine (ATZ) toxicity to natural periphyton in a pulse exposition scenario. Φ_PSII and F_v/F_m were regularly estimated during a 7 h-exposure to each pollutant (0-100 μM) and also later after being transferred in herbicide-free water (up to 36 h). Our results showed that IPU was more toxic than ATZ, but with effects reversible within 12 h. Moreover, these two similarly acting herbicides (i.e. same target site) presented contrasted short term recovery patterns, regarding the previous exposure duration.     

A method of hazard assessment of a gaseous substance with respect to formation of toxic photodecomposition products. Application to CCl4, CCl3F and CCl2F2

CCl₄, CCl₃F and CCl₂F₂ decompose in ambient air forming highly toxic carbonyl halides etc. when exposed to sources of intense uv-emission, e.g. welding arcs. The resulting health hazard is assessed using a mathematical model.