Development of an in vitro toxicological test system based on zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) sperm analysis

The effect of seven heavy metals on the motility parameter of zebrafish sperm was tested in order to develop an in vitro toxicological test system as an alternative to live animal testing. In vitro test systems are currently preferred in ecotoxicology due to their practical and ethical advantages and fish sperm can be a suitable model. A number of studies had been carried out previously on this topic, but the described methods had not been standardized in numerous aspects (donor species, measured endpoint, etc.). In this study, heavy metals (mercury, arsenic, chromium, zinc, nickel, copper, cadmium) were used as reference toxicants with known toxicity to develop a standardized fish sperm in vitro assay. The tested concentrations were determined based on preliminary range finding tests. The endpoints were progressive motility (PMOT, %), curvilinear velocity (VCL, μm/s), and linearity (LIN, %) measured by a computer-assisted sperm analysis (CASA) system. According to our results, PMOT was the most sensitive of the three investigated parameters: dose-response curves were observed for each metal at relatively low concentrations. VCL values were less sensitive: higher concentrations were needed to observe changes. Of the three parameters, LIN was the least affected: dose-response relationship was observed only in the case of mercury (e.g., lowest observed effect concentration (LOEC) of Hg at 120 min: 1 mg/L for PMOT, 2.5 mg/L for VCL, 5 mg/L for LIN; LOEC of Cu at 120 min: 1 mg/L for PMOT, 5 mg/L for VCL, any for LIN). The order of toxicity as determined by PMOT was as follows: Hg²⁺?>?As³⁺?>?Cd²⁺?>?Cu²⁺?>?Zn²⁺?>?Cr³⁺?>?Ni²⁺. In conclusion, we found that PMOT of zebrafish sperm was an accurate and fast bioindicator of heavy metal load. Sperm analysis can be adopted to estimate the possible toxic effects of various chemicals in vitro. Future investigations should concentrate on the applicability of this assay to other contaminants (e.g., organic pollutants).     

Association between environmental exposure to p,p′-DDE and lindane and semen quality

Scientific concern exists about the toxic effect of dichlorodiphenyldichloroethylene (p, p′-DDE) and lindane on male infertility, and the mechanism underlying male reproductive toxicity of this pesticide remains unanswered. We investigated not only the possible association between the chlorinated pesticide levels and semen quality in nonoccupationally exposed men, but also the probable mode of action using mitochondrial membrane potential (MMP), reactive oxygen species (ROS), lipid peroxidation (LPO), and sperm chromatin structure assay (SCSA). A study in 278 men (21–40 years old) who visited Obstetrics and Gynecology Department, KGMU, Lucknow, for semen analysis was conducted. We performed semen analysis according to the WHO guidelines, while p, p′-DDE and lindane analysis was done by the GLC and LPO by the spectrophotometer, and the sperm mitochondrial status, ROS, and SCSA with the flow cytometer. The questionnaire data showed no significant difference in the demographic characteristics between the two groups, i.e., trying to conceive >1 year and proven fertility. However, a significant difference in the concentration of p, p′-DDE and lindane was observed between the groups. When the subjects were divided among four categories by quartile of exposure, the subjects in the highest quartile showed low sperm motility as compared to the subjects in the lowest quartile. Pearson’s correlation showed a significant negative correlation between semen p, p′-DDE, lindane level, and sperm quality and positive association with the number of cells with depolarized mitochondria, elevation in ROS production and LPO, and DNA fragmentation index (DFI). The findings are suggestive that these toxicants might cause a decline in semen quality, and these effects might be ROS, LPO, and mitochondrial dysfunction mediated.     

Reproductive toxicity of lead,cadmium, and phthalate exposure in men

Environmental toxicants viz lead or cadmium and phthalate esters (di(2-ethylhexyl) phthalate [DEHP], dibutyl phthalate [DBP], and diethyl phthalate [DEP]) widely found in different environmental strata are linked to deteriorating male reproductive health. The objective was to assess the relationships between the seminal lead, cadmium, and phthalate (DEHP, DBP, DEP) concentrations at environmental level and serum hormone levels and semen quality in non-occupationally exposed men and specify the effect of individual and combined exposure of toxicants on semen quality. A study of 60 male partners of couples attending the Andrology Laboratory of the Reproductive Biology Department, All India Institute of Medical Sciences (AIIMS), New Delhi, India for semen analysis to assess their inability to achieve a pregnancy was selected for the study. The results of univariate and stepwise multiple regression analysis in the unadjusted model showed a significant correlation between lead or cadmium and phthalates DEHP/DBP/DEP and sperm motility, sperm concentration, and DNA damage. After adjusting for potential confounders, an association with lead or DEHP was only observed. The present data shows that lead (Pb) or cadmium (Cd) or phthalates might independently contribute to decline in semen quality and induce DNA damage. Phthalates might influence reproductive hormone testosterone. These findings are significant in light of the fact that men are exposed to a volley of chemicals; however, due to the small sample size, our finding needs to be confirmed in a larger population.     

Copper and cadmium removal from synthetic industrial wastewater using chitosan and nylon 6

Purpose

Chitosan with nylon 6 membranes was evaluated as adsorbents to remove copper and cadmium ions from synthetic industrial wastewater.

Methods

Chitosan and nylon 6 with glutaraldehyde blend ratio with (1:1+Glu, 1:2+Glu, and 2:1+Glu) have been prepared and these were used as membranes to remove copper and cadmium ions from synthetic industrial wastewater. Characterization of the synthesized membrane has been done with FTIR, XRD, TGA/DTA, DSC, and SEM. Chemical parameters for quantities of adsorption of heavy metal contamination have been done and the kinetics of adsorption has also been carried out.

Results

The optimal pH for the removal of Cd(II) and Cu(II) using chitosan with nylon 6. Maximum removal of the metals was observed at pH 5 for both the metals. The effect of adsorbent dose also has a pronounced effect on the percentage of removal of the metals. Maximum removal of both the metals was observed at 5 g/100 ml of the adsorbent.

Conclusion

Copper and cadmium recovery is parallel at all time. The percentage of removal of copper increased with increase in the pH from 3 to 5. In the case of cadmium containing wastewater, the maximum removal of metal occurred at pH 5. The uptake amount of Cu²⁺ ions on chitosan increased rapidly with increasing contact time from 0 to 360 min and then reaches equilibrium after 360 min; the equilibrium constant for copper and cadmium ions is more or less the same for the adsorption reaction.     

Does static electric field from ultra-high voltage direct-current transmission lines affect male reproductive capacity? Evidence from a laboratory study on male mice

With the development of ultra-high-voltage direct-current (UHVDC) transmission technology and increase in transmission voltage, the issue of environmental static electric field (SEF) pollution is standing out and its possible health effects have caused much public attention. In this study, the effects of chronic exposure to SEF on reproductive capacity of male mice were investigated. Twenty Institute of Cancer Research (ICR) mice were exposed to SEF (56.3 ± 1.4 kV/m, 49 days) generated by a high-voltage device. Several biological end points related to spermatogenesis and testicular function were evaluated, including reproductive organ coefficients, sperm motility and morphology, serum testosterone level, and testicular histology. No significant differences were found between the SEF-exposed and sham-exposed groups at the end of the exposure period. However, further observation through transmission electron microscopy revealed cristae losses in mitochondria of spermatogenic cells after SEF exposure. Nevertheless, the mitochondria injury did not affect sperm motility, which might be explained from the perspective of energy supply. That is, most of the energy required for sperm movement is generated by glycolysis which occurs in the cytoplasm rather than oxidative phosphorylation which occurs in mitochondria. In conclusion, this study indicates that exposure to SEF (56.3 ± 1.4 kV/m, 49 days) has limited effects on male reproductive capacity.     

Efficiency of lipopeptide biosurfactants in removal of petroleum hydrocarbons and heavy metals from contaminated soil

This study describes the potential application of lipopeptide biosurfactants in removal of petroleum hydrocarbons and heavy metals from the soil samples collected from industrial dumping site. High concentrations of heavy metals (like iron, lead, nickel, cadmium, copper, cobalt and zinc) and petroleum hydrocarbons were present in the contaminated soil samples. Lipopeptide biosurfactant, consisting of surfactin and fengycin was obtained from Bacillus subtilis A21. Soil washing with biosurfactant solution removed significant amount of petroleum hydrocarbon (64.5 %) and metals namely cadmium (44.2 %), cobalt (35.4 %), lead (40.3 %), nickel (32.2 %), copper (26.2 %) and zinc (32.07 %). Parameters like surfactant concentration, temperature, agitation condition and pH of the washing solution influenced the pollutant removing ability of biosurfactant mixture. Biosurfactant exhibited substantial hydrocarbon solubility above its critical micelle concentration. During washing, 50 % of biosurfactant was sorbed to the soil particles decreasing effective concentration during washing process. Biosurfactant washed soil exhibited 100 % mustard seed germination contradictory to water washed soil where no germination was observed. The results indicate that the soil washing with mixture of lipopeptide biosurfactants at concentrations above its critical micelle concentration can be an efficient and environment friendly approach for removing pollutants (petroleum hydrocarbon and heavy metals) from contaminated soil.     

Prenatal and postnatal exposure to diazinon and its effect on spermatogram and pituitary gonadal hormones in male offspring of rats at puberty and adulthood

The objective of this research is to study the possible reproductive adverse effects of diazinon on rat offspring exposed in utero and during lactation. Twenty-four Sprague–Dawley female rats (10–12 week old) were randomly assigned to four groups, each consisting of six rats. Group 1 served as the control and these rats were given normal saline orally. Rats in groups 2, 3, and 4 were administered diazinon, dissolved in saline at 10, 15, 30 mg/ kg^?1 body weight, per oral, once daily, during mating, pregnancy and lactation. The male offsprings were examined at puberty and adulthood for body weight, testis weight, epididymis weight, sperm count, motility and morphology, pituitary-gonadal hormone levels. At 30 mg kg^?1 dose, the male offsprings showed a decrease in testicular weight, sperm count, motility, with an increase in abnormal sperm percentage and a decline in pituitary-gonadal hormones, at puberty. Upon attaining adulthood, there was a decrease in testicular weight, sperm count and motility with an increase in abnormal sperm percentage and a decrease in pituitary hormone level. There was evidence of some adverse reproductive effects on the male offspring at the 15 mg/ kg^?1 dose. Most of the adverse effects were irreversible and were evident at both puberty and adulthood in the offsprings, although a few parameters reverted to the normal growth pattern. Diazinon is a reproductive toxicant for male offsprings if exposed during prenatal and postnatal phases.     

Effects of chronic exposure to lead,copper, zinc,and cadmium on biomarkers of the European eel,Anguilla anguilla

Exposure to specific metallic compounds can cause severe deleterious modifications in organisms. Fishes are particularly prone to toxic effects from exposure to metallic compounds via their environment. Species that inhabit estuaries or freshwater environments can be chronically affected by persistent exposure to a large number of metallic compounds, particularly those released by industrial activities. In this study, we exposed yellow eels (European eel, Anguilla anguilla) for 28 days to environmentally relevant concentrations of four specific metals; lead (300, 600, and 1,200 μg/l), copper (40, 120, and 360 μg/l), zinc (30, 60, and 120 μg/l) and cadmium (50, 150, and 450 μg/l). The selected endpoints to assess the toxicological effects were neurotransmission (cholinesterasic activity in nervous tissue), antioxidant defense, and phase II metabolism (glutathione-S-transferase [GST] activity, in both gills and liver tissues), and peroxidative damage. The results showed an overall lack of effects on acetylcholinesterase for all tested metals. Lead, copper, and cadmium exposure caused a significant, dose-dependent, increase in GST activity in gill tissue. However, liver GST only significantly increased following zinc exposure. No statistically significant effects were observed for the thiobarbituric acid reactive substances assay, indicating the absence of peroxidative damage. These findings suggest that, despite the occurrence of an oxidative-based response after exposure to lead, copper, and cadmium, this had no consequence in terms of peroxidative membrane damage; furthermore, cholinergic neurotoxicity caused by lead, copper, and cadmium did not occur. The implications of these results are further discussed.     

Ultrasonic vibration seeds showed improved resistance to cadmium and lead in wheat seedling

Heavy metals have long-term adverse impacts on the health of soil ecosystems and even exhibit hazardous influences on human health. Literatures have shown that heavy metals could result in the reduction of crops growth and development and finally result in crops production decline. To determine whether or not ultrasonic vibration alleviate damage induced by cadmium and lead in crops, the wheat seeds, which is one of the most important agriculture crops in China and other countries in the world, were exposed to 10 min ultrasonic vibration and then the toxicological effects were investigated. Wheat seeds were soaked for 3 h with water and then the seeds were placed in clean beaker with some water, the beaker were placed in ultrasonic apparatus to vibrate (model, KQ-200VDV; frequency, 45 KHz; power, 160 W). Pretreatment seeds of 80 were sown in dishes (Ø 15 cm). After seeds emergence, the seedlings were thinned to 60 per dish. The dishes with seedlings were placed in a growth chamber maintained at 25 °C, 70 % relative humidity and 380 μmol?mol^?1 CO₂ under dark condition. A 400 μmol?m^?2?s^?1 photosynthetically active radiation was provided for 8 h (dark for 16 h) after the seed germination. When the seedlings were 2 days old, the seedlings were subjected to cadmium and lead for 4 days and then some selective biochemical and physiological parameters were measured. (1) Although each doses of ultrasonic vibration could improve seed germination, enhance biosynthesis of protein and chlorophyll and seedlings growth, the optimum dosage of ultrasonic vibration was 10 min. (2) Compared with the controls, cadmium and lead stress led to significant increase in the concentrations of malondialdehyde (MDA) and O^?2 and in the conductivity of electrolyte leakage, but the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), the glutathione concentration, and the shoot weight were decreased by Cd and Pb stress. In the case of the seeds exposed to ultrasonic vibration and the seedlings followed by cadmium lead stress, the concentrations of MDA and O^?2, and the conductivity of electrolyte leakage were significantly lower than those in cadmium and lead stress; the activities of CAT, SOD, and GR and the shoot weight were significantly higher (except for glutathione (GSH) concentration) than those in cadmium and lead stress seedlings. The membrane is responsible for the selective inflow and outflow of molecules, ions, and water, and is a dynamic structure that performs a variety of functions. Cellular membrane systems play an important role in the compartmentalization of cells and maintaining intercellular homeostasis. Abiotic and biotic stress can induce functional impairments to the cellular membrane systems through triggering an increased formation of reactive oxygen species (ROS), such as superoxide (O₂ ^?), hydrogen peroxide (H₂O₂), and hydroxyl radicals. There are several pathways that can be utilized to eliminate ROS in plants, e.g., CAT, SOD, and GR and GSH, etc. compared with controls, cadmium, and lead enhanced the concentrations of ROS; decreased the SOD, CAT, and GR activities; the GSH concentration, and the seedling growth. In the case of ultrasonic pretreatment followed by cadmium and lead stresses, the activities of CAT, SOD, and GR were significant higher, and the conductivity of electrolyte leakage and the concentrations of MDA and O₂ ^? were significant lower than that of those subjected by cadmium and lead stress. This phenomenon demonstrated ultrasonic pretreatment can help plant eliminate the ROS by enhance the activities of antioxidant enzymes. These results suggested that ultrasonic vibration can alleviate the toxicological effect induced by heavy mental.     

Cadmium in blood of Tunisian men and risk of bladder cancer: interactions with arsenic exposure and smoking

Prior investigations identified an association between low-level blood arsenic (As) and bladder cancer risk among Tunisian men but questions remain regarding confounding by cadmium (Cd), a well-established bladder carcinogen. A case–control study of Tunisian men was re-examined to assess the levels of cadmium in blood and reparse the association between the simultaneous exposure to these metals and bladder cancer risk. Levels of blood Cd were significantly twice higher among cases than in controls (P?<?0.05) and were positively correlated with smoking and age. Additionally, analysis of metal levels among non-smokers according to the region of residence showed very high blood Cd and As levels for the coastal regions of Sfax and central Tunisia. After controlling for potential confounders, for low blood As levels (<0.67 μg/L), the OR for blood Cd was 4.10 (95 % CI 1.64–10.81), while for higher levels (>0.67 μg/L), it was reduced to 2.10 (CI, 1.06–4.17). Adjustment for Cd exposure did not alter the risk associated to As exposure. This study is the first to report the relationship between Cd exposure and risk of bladder cancer occurrence in interaction with smoking and As exposure. Smoking is shown to be the main exposure source to Cd in the Tunisian population but also environmental pollution seems to be responsible of Cd exposure among non-smokers. Exposure assessment studies encompassing a wider population are needed.     

Effect of Isoflavones on Reproductive Performance,Testosterone Levels,Lipid Peroxidation,and Seminal Plasma Biochemistry of Male Rabbits

Abstract

The objective of this study was to determine the effect of either 2.5 mg/kg Body Weight or 5 mg/kg Body Weight (BW) doses of isoflavones on semen quality, testosterone levels, lipid peroxidation and semen biochemistry of male New Zealand White rabbits. Animals were given both 2.5 mg/kg BW and 5 mg/kg BW doses of isoflavones. The tested doses were given to rabbits orally every other day for 13 weeks. Treatment with isoflavones caused an increase (p < 0.05) in libido (by decreasing the reaction time), sperm concentration, sperm motility (%), total motile sperm per ejaculate (TMS), packed sperm volume (PSV), total functional sperm fraction (TFSF), total sperm output, initial fructose concentration and normal sperm, while dead sperm was reduced compared to control animals. On the other hand, ejaculate volume, initial hydrogen ion concentration (pH) and plasma testosterone levels did not change in treated animals with both doses of isoflavones as compared to control. Concentrations of thiobarbituric acid-reactive substances (TBARS), total lipids, and low density lipoprotein were significantly (p < 0.05) reduced in seminal plasma of rabbits treated with either 2.5 mg/kg BW or 5 mg/kg BW doses of isoflavones. While, the activities of glutathione S-transferase (GST), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), acid phosphatase (AcP), and alkaline phosphatase (AlP) were significantly (p < 0.05) increased in seminal plasma of treated animals. Also, total cholesterol, percentage cholesterol (out of total lipids), and high density lipoprotein were significantly (p < 0.05) increased, while triglyceride did not change in seminal plasma of treated animals. Supplementation at either level of isoflavones did not cause changes in live body weight (LBW), dry matter intake (DMI), and relative weights of testes and epididymis. The present results showed that either 2.5 mg/kg BW or 5 mg/kg BW doses of isoflavones caused an improvement of some semen characteristics and did not have negative effects on male fertility.     

Protective Role of Isoflavones Against the Toxic Effect of Cypermethrin on Semen Quality and Testosterone Levels of Rabbits

Cypermethrin is considered as one of the endocrine disruptors. Isoflavones play an important role in various physiological processes in the body. It has both estrogenic and antioxidant effects. The objective of this study was to evaluate the protective role of isoflavones (2 mg/kg B.W) on semen quality and plasma testosterone levels of male New Zealand White rabbits given sublethal dose (24 mg/kg BW every other day for 12 weeks) of cypermethrin. Results showed that treatment with cypermethrin caused a significant decreases (P < 0.05) in ejaculate volume, sperm concentration, total sperm output, sperm motility (%), total motile sperm per ejaculate (TMS), packed sperm volume (PSV), semen initial fructose and plasma testosterone. In addition, live body weight (LBW), dry matter intake (DMI) and relative weights of testes and epididymis were decreased. On the other hand, treatment with cypermethrin increased (P < 0.05) the numbers of abnormal and dead sperms, and initial hydrogen ion concentration (pH). Results indicated that the presence of isoflavones together with cypermethrin was capable to minimize its harmful effects. Treatment with isoflavones alone had positive effects on some semen characteristics in spite of it is considered as estrogen‐like compound. Since it causes significant increases in libido (by decreasing the reaction time), PSV, sperm motility and TMS, while abnormal and dead sperm were reduced compared to control animals. Meanwhile, isoflavones had no negative effect on ejaculate volume, total sperm output, sperm concentration, initial fructose concentration, pH and plasma testosterone levels. Results demonstrated the beneficial influences of isoflavones in reducing the negative effects of cypermethrin on reproductive characteristics of mature male rabbits. Interestingly, data showed that isoflavones alone caused an improvement in some semen quality and had no negative effects on male fertility, and did not have negative effects on male fertility.     

Enzymatic biomarkers as indicators of dietary cadmium in gypsy moth caterpillars

Heavy metals damage the structure, chemistry, and function of cells, including enzyme systems inside them. Variation in the profile of biochemical biomarkers in prevalent species should be used for assessing environmental contamination. The present study pays attention to the phosphatases present in the midgut of gypsy moth fourth instar caterpillars, which had been exposed to short- and long-term cadmium intake at 10 and 30 μg Cd/g dry food. Chronic cadmium ingestion significantly inhibited the activity of all examined phosphatases, while only the activity of lysosomal phosphatase was acutely decreased. Total acid phosphatase activity recovered from both long-term cadmium treatments within 3 days. The low index of phenotypic plasticity was connected to high variability of plasticity. Dependence of phosphatase isoforms on genotype and duration of cadmium treatment was determined. We concluded that, with further investigations, profiling of total acid phosphatase activity, as well as the lysosomal fraction can be used as a biomarker for acute sublethal metal toxicity.     

Body metal concentrations and glycogen reserves in earthworms (Dendrobaena octaedra) from contaminated and uncontaminated forest soil

Stress originating from toxicants such as heavy metals can induce compensatory changes in the energy metabolism of organisms due to increased energy expenses associated with detoxification and excretion processes. These energy expenses may be reflected in the available energy reserves such as glycogen. In a field study the earthworm, Dendrobaena octaedra, was collected from polluted areas, and from unpolluted reference areas. If present in the environment, cadmium, lead and copper accumulated to high concentrations in D. octaedra. In contrast, other toxic metals such as aluminium, nickel and zinc appeared to be regulated and kept at low internal concentrations compared to soil concentrations. Lead, cadmium and copper accumulation did not correlate with glycogen reserves of individual worms. In contrast, aluminium, nickel and zinc were negatively correlated with glycogen reserves. These results suggest that coping with different metals in earthworms is associated with differential energy demands depending on the associated detoxification strategy.     

Characterization of contaminants and evaluation of the suitability for land application of maize and sludge biochars

Prior to the application of biochar as an agricultural improver, attention should be paid to the potential introduction of toxicants and resulting unintended impacts on the environment. In the present study, the concentrations of polycyclic aromatic hydrocarbons (PAHs), heavy metals, and mineral elements were determined in maize and sludge biochars produced at 100 °C increments between 200 and 700 °C. The concentration ranges of total PAHs were 358–5,136 μg kg^?1 in maize biochars and 179–70,385 μg kg^?1 in sludge biochars. The total heavy metals were detected at the following concentrations (mg kg^?1): Cu, 20.4–56.7; Zn, 59.7–133; Pb, 1.44–3.50; Cd, <0.014; Cr, 8.08–21.4; Ni, 4.38–9.82 in maize biochars and Cu, 149–202; Zn, 735–986; Pb, 54.7–74.2; Cd, 1.06–1.38; Cr, 180–247; Ni, 41.1–56.1 in sludge biochars. The total concentrations of PAHs and heavy metals in all maize biochars and most sludge biochars were below the control standards of sludge for agricultural use in China, the USA, and Europe. The leachable Mn concentrations in sludge biochars produced at below 500 °C exceeded the groundwater or drinking water standards of these countries. Overall, all the maize biochars were acceptable for land application, but sludge biochars generated at temperatures between 200 and 500 °C were unsuitable for application as soil amendments due to their potential adverse effects on soil and groundwater quality.     

Dental enamel as biomarker for environmental contaminants in relevant industrialized estuary areas in São Paulo,Brazil

Heavy metal contamination is a long-standing and very well-known public health problem, and its exposure can cause damage to several organs of human body, especially on the central nervous system of young children and teenagers. The aim of this article is to evaluate lead, cadmium, and manganese contamination in 125 children from 6 to 13 years old living in contaminated areas during the period from 2006 to 2009 (São Vicente, Cubatão Downtown, Bertioga and Cubatão Pilões/Água Fria). This estuary area is the most important example of environmental degradation by chemicals from industrial sources. This is a cross-sectional study through clinical examinations and dental enamel tests. All mothers from these children lived in the area since before the pregnancy. Lead, cadmium, and manganese levels (μg/g) were measured on dental enamel samples through graphite furnace atomic absorption spectrometry, searching for the occurrence of heavy metals. The mean lead concentrations were 139.48 μg/g in Cubatão Pilões/Água Fria, 170.45 μg/g in Cubatão Downtown, 213.52 μg/g in São Vicente, and 151.89 μg/g in Bertioga. The mean cadmium concentrations were 10.83 μg/g in Cubatão Pilões/Água Fria, 12.58 μg/g in Cubatão Downtown, 10.92 μg/g in São Vicente, and 14.57 μg/g in Bertioga. The mean manganese concentrations were 23.49 μg/g in Cubatão Pilões/Água Fria, 30.90 μg/g in Cubatão Downtown, 41.46 μg/g in São Vicente, and 42.00 μg/g in Bertioga. Dental surface enamel may be used as an efficient biomarker of past environmental exposure to lead, manganese, and cadmium which are associated to well-known sources of heavy metal contamination. The results suggest that the evaluated children were exposed to sources of lead, cadmium, and manganese since before their conceptions. Although Bertioga initially was chosen as a control area of this study, it was also was verified to have heavy metal contamination on examined children.     

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+.     

The effect of pesticides and metals on acetylcholinesterase (AChE) in various tissues of blue mussel (Mytilus trossulus L.) in short-term in vivo exposures at different temperatures

The purpose of the study was to assess the impact of short-term exposure to selected toxicants as well as metal accumulation upon acetylcholinesterase (AChE) in the blue mussel, Mytilus trossulus L., in laboratory in vivo experiments. Mussels were exposed for up to 48 hours to a mixture of copper (Cu^{2 +}, 400 μ g L^?1) and cadmium (Cd^{2 +}, 200 μ g L^?1), to dichlorvos (DDVP, 100 μ g L^?1), and to carbaryl (100 μ g L^?1) at two temperatures: 5°C and 20°C. Samples were collected after 0, 6, 12, 24, and 48 hours of exposure, and AChE activity and metal concentration (where applicable) were analysed in gills, digestive gland, mantle+muscles, and the whole soft tissue. Very strong AChE inhibition was observed in response to the dichlorvos treatment, mainly in gills. Carbaryl and the metals caused a short-term inhibition effect. Considerable differences in AChE activity between the two temperatures were noticed. In particular, the metals were accumulated much faster at 20°C than at 5°C, especially in gills. No correlation between AChE activity and metal concentration was found. Gills turned out to be the optimal tissue for AChE activity analysis in short-term studies.