Milk transfer and tissue uptake of mercury in suckling offspring after exposure of lactating maternal guinea pigs to inorganic or methylmercury

Maternal guinea pigs were injected with mercuric chloride (HgCl₂; 1 mg Hg/kg body weight) or methylmercury (MeHg; 1 mg Hg/kg) 12 h after parturition, and exposure of the offspring to mercury (Hg) via breast milk were studied on days 3, 5 and 10 postpartum. Milk Hg concentrations were lower than maternal plasma Hg concentrations regardless of the form of Hg given to the dams. Milk Hg was higher in HgCl₂-treated dams than in MeHg-treated dams. In MeHg-treated dams, MeHg was separately determined. While the ratio of MeHg to T-Hg decreased in the dams’ plasma, it did not in the milk. There was a strong correlation between milk and plasma T-Hg concentrations in HgCl₂ treated dams. In the milk of MeHg-treated dams, the plasma MeHg concentrations correlated better than did the plasma T-Hg concentrations. In the offspring, regardless of the chemical forms of Hg given to the dams, the highest Hg concentrations were found in the kidney, followed by the liver and the brain. Brain Hg concentrations were, however, significantly higher in the offspring of MeHg-treated dams than in those of HgCl₂-treated dams. In addition, Hg levels in the major organs of the offspring of HgCl₂-treated dams peaked on day 5 postpartum, while those of MeHg-treated dams did not show a significant decrease up to day 10 postpartum. These facts indicate that the two chemical forms of Hg were transferred to the offspring via the breast milk and were distributed differently, depending on the chemical form, to the offspring’s tissues.     

Selenium concentrations in brain after exposure to methylmercury: relations between the inorganic mercury fraction and selenium

 Three groups of female monkeys (Macaca fascicularis) were exposed to methylmercury (MeHg, p.o. 50 μg Hg/kg body wt per day) for 6, 12, or 18 months. One group was exposed to MeHg for 12 months and kept unexposed for 6 months before sacrifice. Another group of three monkeys was exposed to HgCl₂ i.v. for 3 months. Total and inorganic mercury concentrations in occipital pole and thalamus were determined by cold vapor atomic absorption spectroscopy. Selenium concentrations were analyzed by hydride generation atomic absorption spectroscopy. The results indicated an association between concentrations of inorganic mercury and selenium in both occipital pole and thalamus in the MeHg-exposed animals. A linear regression model using concentrations of inorganic mercury (nmol/g wet wt) as independent variable, and selenium concentrations (nmol/g wet wt) as the dependent variable showed significant correlations between the variables in both occipital pole and thalamus (r = 0.85 and r = 0.91, P <0.0001). The intercept of the regression line was slightly lower (about 2 nmol Se/g wet wt) than the selenium concentrations found in control monkeys (about 3 nmol Se/g wet wt). There was a tendency to a “hockey stick”-shaped relationship between concentrations of selenium and inorganic mercury in the thalamus of monkeys with ongoing exposure to MeHg. An important role for selenium in the retention of mercury in brain is indicated. Received: 6 April 1994 / Accepted: 5 October 1994     

Methylmercury transport across the placenta via neutral amino acid carrier

 Methylmercury (MeHg) penetrates the placental barrier to affect developing fetuses in the uterus. However, the mechanism of placental MeHg transport is not well defined. To clarify the MeHg transport system that functions in the placenta, pregnant rats were intravenously administered MeHg on day 18 of gestation. The fetal blood was collected from the umbilical cord at 30 and 60 min after the administration, and its mercury concentration was measured. MeHg was found to be rapidly transported to the fetal blood in a time- and dose-dependent manner, and predominantly distributed in the blood cells there. MeHg transport was effectively suppressed by the co-injection of neutral amino acids, i.e., L-methionine and L-phenylalanine, suggesting that MeHg is actively transported as its cysteine conjugate via the neutral amino acid carrier system. The suppression by methionine was not so marked as by phenylalanine. Since methionine administration caused a rapid increase of the cysteine, which functioned as a predominant carrier in MeHg transport, in the maternal plasma, newly synthesized cysteine seemed to accelerate the mercury uptake. Accordingly, the acceleration by the extra cysteine would compensate partly the competitive effect of methionine as a neutral amino acid. Received: 20 June 1995/Accepted: 21 August 1995     

Postnatal behavioral effects in mice after prenatal exposure to methylmercury

CFW mice were injected with methylmercury hydroxide (1, 2, 3, 5 or 10 mg/kg as mercury) on Day 8 of gestation. Mice treated with 3, 5 or 10 mg/kg averaged $\text{1}\text{3}$ fewer pups than controls. Pups from these treated animals weighed less than controls and the weight differences persisted through weaning but were no longer significant at 56 days of age. Mice exposed to methylmercury in utero showed significant differences from controls in their behavior in a 2-way active avoidance shuttle box and in a punishment situation but not when tested in an open field, a water escape runway or a conditioned suppression paradigm. Neither the mothers nor progency of the mice exposed prenatally to methylmercury showed behavioral deficits.     

The effect of various dietary fibres on tissue concentration and chemical form of mercury after methylmercury exposure in mice

The whole-body retention of mercury after exposure of BALB/c mice to methylmercury was measured in animals fed fibre-free, 5% pectin, 5% cellulose or 5, 15 or 30% wheat bran diets. The rate of elimination of mercury was dependent on the diet fed, with dietary bran increasing the rate of elimination. The incorporation of 15 or 30% bran in the diet of the mice decreased the total mercury concentration in the brain, blood and small intestine, although the effects were significant only in those animals on 30% bran diet. The fibres had little effect on mercury levels in other tissues. The proportion of mercury found in the mercuric form was significantly greater in liver, kidneys and gut of mice fed bran. The results suggest that dietary bran may reduce the levels of mercury in the brain after methylmercury exposure and may therefore reduce the neurotoxic effects of the organomercurial. We suggest that wheat bran exerts its effects on mercury retention and brain level via a modification of the metabolic activity of the gut microflora.     

Developmental disturbances of the fetal brain in guinea-pigs caused by methylmercury

Pregnant guinea-pigs of Hartley strain were orally administered methylmercuric chloride once at a dose of 7.5 mg Hg/animal (weighing 500–800 g) on one of days 21, 28, 35, 42 or 49 (3–7 weeks) of gestation. They were killed on day 63 (9 weeks) and their fetuses were removed. Both maternal and fetal blood, brain, liver and kidney, and fetal hair, urine, gastric content and amniotic fluid as well, were sampled for mercury analysis. The fetal brains were also examined pathologically. The maternal kidney contained mercury at a high concentration but the fetal kidney did not. The mercury concentration was strikingly high in the fetal hair, but fairly low in the urine, gastric contents and amniotic fluid. Mercury distributed unevenly in various brain regions of both dams and fetuses after treatment at 6 and 7 weeks of pregnancy (3 and 2 weeks before sampling). The concentration was high in the neopallium and archipallium, followed by the paleopallium, diencephalon and mesencephalon, but low in the rhombencephalon, including cerebellum. Mercury contents were relatively low and distributed almost evenly in various brain regions of both the dams and fetuses following treatment at 3, 4, and 5 weeks of pregnancy. Morphologically, the fetal brains were disturbed in the development following treatment at 3, 4 and 5 weeks of pregnancy. The cerebral cortex was thinned, the nucleus caudatus putamen and the hippocampal formation were reduced in size, and the lateral ventricles were dilated. However, the histological architecture of the cerebral cortex was not strikingly maldeveloped; only a slight disarrangement of the cellular alignment was noted. Following treatment at 6 and 7 weeks of pregnancy, focal degeneration of the neuronal cells was observed in the fetal neocortex; the severe cases showed spongy degeneration and dysgenetic hydrocephalus.     

Acute effects of methylmercury on hepatic and renal glutathione metabolisms in mice

Because of its high affinity to the sulfhydryl group, the in vivo fate of methylmercury (MeHg) is closely related to the glutathione (GSH) metabolism. Here, to examine the possible effects of MeHg on the GSH metabolism, C57BL female mice were challenged by this heavy metal at a marginal dose level to induce slight renal dysfunction. Liver and blood GSH levels decreased by 16% and 20%, respectively, 24 h after MeHg (160 μmol/kg) administration, whereas kidney and plasma levels drastically increased. The GSH half-lives obtained using L-buthionine-(S,R)-sulfoximine were shortened by 17% in the liver, but lengthened by 28% in the kidney. The accelerated secretion of GSH from the liver and/or blood cells might have caused increased plasma levels of the tripeptide, which in turn could increase the supply of the constituent amino acids for GSH synthesis to the kidney. Furthermore, renal γ-glutamylcysteine synthetase activity, a rate-determining enzyme in GSH biosynthesis, was found to be enhanced in the MeHg-treated group. The marked increase in the renal GSH levels induced by MeHg could be due to the increased synthesis and the decreased efflux of the tripeptide in this tissue. The MeHg-induced alterations of GSH metabolism described here might reflect one of the defense mechanisms of bioorganisms against the challenge by MeHg. Received: 29 November 1993 / Accepted: 25 April 1994     

甲基汞对大鼠的行为致畸效应研究

目的探讨妊娠期甲基汞暴露对Wistar大鼠的母体毒性及仔代的行为致畸效应.方法 Wistar孕鼠80只于妊娠第6～9天采用甲基汞0.00、0.01、0.05和2.00mg·kg-1@d-1连续灌胃染毒.分别进行母体毒性、胚胎毒性、仔鼠早期生理发育和神经行为发育指标、仔鼠迷宫和程序控制行为测试、亲仔两代大鼠脑组织形态学观察和单胺类神经递质(去甲肾上腺素、多巴胺、5-羟色胺)的测定.整个实验采用双盲法.结果未观察到明显的母体毒性;3个剂量组胎仔的体重、尾长均低于对照组(P＜0.01);各剂量组仔鼠的体重增长、早期生理及神经行为发育滞后于对照组(P＜0.05);各剂量组仔鼠迷宫错误次数均比对照组多(P＜0.05),具有剂量-效应关系(rs=0.257,P＜0.05);程序控制行为学习成绩比对照组降低(P＜0.05),有剂量-效应关系(rs=-0.727 3,P＜0.01);各剂量组母鼠和仔鼠脑组织均未见形态学改变,但脑组织单胺类神经递质含量均比对照组明显增高(P＜0.05),有剂量-效应关系(s=0.712 4～0.925 7,P＜0.01).结论甲基汞在不引起可观察到母体毒性剂量下,就可产生胚胎毒性,影响仔鼠神经系统的发育,导致神经行为功能的改变.     

Mercury and methylmercury contamination in Mytilus galloprovincialis from Taranto Gulf (Ionian Sea,Southern Italy): Risk evaluation for consumers

Total mercury (THg) and methylmercury (Me-Hg) concentrations were determined in mussels (Mytilus galloprovincialis) from 10 stations located in the Mar Piccolo of Taranto (Ionian Sea, Taranto Gulf) an important semi-enclosed basin in Italy, devoted to mussel culture activities. The obtained results show that THg and Me-Hg concentrations ranged from 0.236 to 0.559 μg g⁻¹ d.w. and from 0.066 to 0.155 μg g⁻¹ d.w., respectively. Consequently, the Me-Hg/THg ratios ranged from 17% to 49%. The dietary intake of THg and Me-Hg were studied among children and adults from Taranto (Southern Italy). The estimated weekly intake for THg and Me-Hg was below the Provisional Tolerable Weekly Intake (PTWI) established by European Food Safety Autority (EFSA) for all sampled mussels, though their consumption provides a THg intake in children near the PTWI.     

Impact of a risk-benefit advisory on fish consumption and dietary exposure to methylmercury in France

We designed the CORAI (COnsumer Risk Advisory Inquiry) study to observe consumer reactions' after an advisory revealing risk of methylmercury contamination together with benefits of Long-Chain Poly Unsaturated Fatty Acids of the n-3 variety (LC n-3 PUFA). The message was very close to the ones commonly delivered by national food agencies and included recommendations for women of childbearing age and children below 15 years old. Two groups of subjects including consumers at risk were selected. Participants recorded the frequency of their fish consumption detailed by species for them and their family over a one-month period one month before, a month immediately after and 3 month after the advisory. Results were compared between consumers receiving the advisory and controls. Results show that the message revelation led to a significant decrease in total fish consumption which is greater for children below 6 years old than for the children between 6 and 15 years old and women. The consumption of the most contaminated fish quoted in the advisory, rarely consumed and poorly known by French consumers did not decrease in any group despite the advice to avoid their consumption. The consumption of other fish products quoted in the advisory but frequently consumed and better known, as canned tuna, did decrease and was a major contributor to the overall reduction of exposure for the advised group. Before the information, about 3% of women of childbearing age are exceeding the PTWI for MeHg and both the average and the high percentiles of the exposure to MeHg are decreasing significantly in the advised group. Regarding the number of subjects of the advised group exceeding the PTWI, they were 6, 3 and 2, respectively, in May, June and September. Accompanying questionnaires show that consumers imperfectly memorize most of the fish species quoted in the recommendation. This paper concludes that consumer advisory, which is a major tool for risk management, has a minimal effect under our experimental conditions to reduce the exposure of groups at risk. Messages to be carried to consumers should be carefully tested for long term memorization in order to become more effective.     

Effects of postnatal exposure to methylmercury on spatial learning and memory and brain NMDA receptor mRNA expression in rats

The extreme vulnerability of developing nervous system to methylmercury (MeHg) is well documented. Still unclear is the consequence of different postnatal period exposure to MeHg. We investigated the critical postnatal phase when MeHg induced neurotoxicity in rats and the underlying mechanism. Rats were given 5 mg/(kg day) methylmercury chloride (MMC) orally on postnatal day (PND) 7, PND14, PND28, and PND60 for consecutive 7 days. A control group was treated with 0.9% sodium chloride solution 5 ml/(kg day) instead. On PND69, spatial learning and memory was evaluated by Morris water maze test. Behavior deficits were found in MMC-treated rats of PND7 and PND14 groups (p < 0.01). N-methyl-d-aspartate (NMDA) receptor 2 subunits mRNA expressions were evaluated 3 days after the last administration. In hippocampus, the mRNA expression of NR2A and NR2B decreased, but the NR2C expression increased in PND14 group following MMC-treatment (p < 0.01). In cerebral cortex, mRNA expression of NR2A decreased, with NR2C expression elevating in PND14 group following MMC-treatment (p < 0.05). These observations suggest that the postnatal exposure to MeHg during PND7–20 could cause neurobehavioral deficits which extend to adulthood. Furthermore, the abnormal expression of NMDAR 2 subunits might associate with the impairment.     

Effects of early life exposure to methylmercury in Daphnia pulex on standard and reduced food ration

As a well-known eco-toxicological model organism, Daphnia pulex may also offer advantages in human health research for assessing long-term effects of early life exposures to coupled stressors. Here, we examine consequences of early life exposure to methylmercury (MeHg) under standard and reduced food ration. We exposed Daphnia for 24 h in early life to varying concentrations of methylmercury(II) chloride (0, 200, 400, 800 and 1600 ng/L) and thereafter kept Daphnia on either a standard or a reduced food ration. The data suggests an additive effect of MeHg concentration and food ration on decreasing lifespan, although MeHg concentration does not affect survival linearly. Food ration and MeHg concentration were predictive of reduced reproduction, and there is some evidence of an interaction (p = 0.048). Multi-stressor work in alternative model systems may be useful for prioritizing research, taking into account potential antagonistic, additive or synergistic effects that nutritional status may have on chemical toxicity.     

Prenatal methylmercury exposure affects spatial vision in adult monkeys

Decades of research have demonstrated that exposure to methylmercury (MeHg), a ubiquitous environmental pollutant, can have both early and long-term neurobehavioral consequences in exposed offspring. The present study assessed visual functioning in adult macaque monkeys (Macaca fascicularis) exposed in utero to 0, 50, 70, or 90 microg/kg/day of MeHg hydroxide. Twenty-one full-term, normal birth weight offspring (9 controls, 12 exposed) were tested at approximately 11-14.5 years of age on a visual contrast sensitivity task. A forced-choice tracking procedure was utilized with spatial frequencies of 1, 4, 10, and 20 cycles per degree of visual angle. On each test session, a single spatial frequency was presented across five levels of contrast, each differing by 3 dB. Methylmercury-exposed monkeys exhibited reduced contrast sensitivity thresholds, particularly at the higher spatial frequencies. The degree of visual impairment was not related to MeHg body burden or clearance and almost half of the exposed animals were unimpaired. The results from this study demonstrate that chronic in utero MeHg exposure, at subclinical levels, is associated with permanent adverse effects on spatial vision in adult monkeys.     

Chronic exposure to cadmium attenuates behavioral sensitization to morphine

 The purpose of this investigation was to assess the impact of dietary cadmium on morphine-induced changes in locomotor activity. Adult male rats were exposed ad libitum to an adulterated food supply containing 100 ppm added cadmium chloride, or an identical diet containing no added cadmium, for 45 days prior to testing for the locomotor activating effects of successive daily morphine administration (0, 5,10, or 20 mg/kg per session) on locomotor activity. On day 1 of testing, increasing doses of morphine produced a dose-related suppression of activity, and this sedative effect was greater in control than in cadmium-exposed animals. Repeated morphine administration resulted in tolerance to the sedative effects of the drug, and a systematic elevation of locomotor activity over the 14-day testing period was observed, with the augmentation (sensitization) effect more pronounced in control than cadmium-exposed animals. There was no indication that conditioning (context) events played a role in the effects observed here. Received: 23 May 1996 / Final version: 6 February 1997     

Inhibitory effect of methylmercury on migration and tube formation by cultured human vascular endothelial cells

 The effect of methylmercury chloride (MeHg) on migration and tube formation by cultured human umbilical vein endothelial cells (HUVECs) was quantitatively analyzed. The distance of endothelial cell outgrowth from the scraped edge of a monolayer was measured. HUVEC outgrowth was inhibited by MeHg (1.0–5.0 μM) treatment in a dose-dependent manner. Tube formation was studied by culturing the cells on gelled basement membrane matrix (Matrigel). Treatment of HUVECs with 0.1–5.0 μM MeHg for 24 h inhibited tube formation dose-dependently. These results suggest that migration and tube formation by HUVECs are susceptible to MeHg cytotoxicity, and that MeHg could be injurious to endothelial cell function, which may be involved in the pathogenesis of arteriosclerosis. Received: 29 July 1994 / Accepted: 22 November 1994     

A longitudinal analysis of prenatal exposure to methylmercury and fatty acids in the Seychelles

Background

Maternal fish consumption during pregnancy exposes the fetus simultaneously to methylmercury (MeHg) and long chain polyunsaturated fatty acids (LCPUFA). Data from the Seychelles Child Development Nutrition Study (SCDNS) showed a negative association of MeHg with child development when children were 30 months of age, only when controlling for LCPUFA. Concomitantly, n − 3 LCPUFA were found to have a significant positive association only at 9 months. These findings suggest that the effects of MeHg and LCPUFA may vary with age over the first few years of life. We address this by including outcomes at two ages and adjusting for the child's age at testing.

Methods

A longitudinal analysis utilizing linear mixed models was performed to assess the associations of maternal hair total mercury (THg, a biomarker for MeHg) and maternal LCPUFA with children's Bayley Scales of Infant Development Psychomotor Developmental Index (BSID-II PDI) at 9 and 30 months of age, and to determine whether these associations change over time. Data from 228 children were included.

Results

Maternal hair MeHg had a negative effect on BSID PDI, while maternal n − 3 LCPUFA had a positive effect. These effects did not change significantly from 9 to 30 months in this analysis.

Conclusions

The longitudinal analysis provides increased power for estimating the relationships of prenatal MeHg and LCPUFA exposures during child development. Significant associations of these exposures in opposite directions confirm the importance of LCPUFA in development and the need to adjust for maternal nutrition when studying prenatal MeHg exposure.     

d-Amphetamine unmasks postnatal consequences of exposure to methylmercury in utero: methods for studying behavioral teratogenesis

Pregnant rats were given various doses of methylmercury (MM) at three different stages of gestation (Days 0, 7 or 14). Administration of 56% or 27% of the dose given on the first day (Day 0) of pregnancy to 7 or 14 day pregnant rats, respectively, resulted in equivalent concentrations of MM ni 19 day old feti and 1 day and 1 week old neonates. A single, 5 mg MM/kg, oral dose on Day 0, or its equivalent on Days 7 or 14 of gestation did not produce any signs of toxicity in pregnant dams or their offspring. Weight gain of pregnant dams, litter size, litter weights at birth or at weaning and gross physical appearance were not different amongst the various treated groups and their respective controls. Operant level of bar pressing and acquisition of a discrete trial autoshape task indicated no differences with respect to operant levels, rate of acquisition or asymptotic performance resulting from exposure to MM in utero. The operant (autoshaped) behavior showed sex related differences to the disrupting influence of d-amphetamine (d-A); females were significantly more sensitive than males. Moreover, both males and females whose dams were treated with MM on Day 0 or 7 of pregnancy were significantly less affected by the d-A when compared with controls. Offspring born to dams given MM on Day 14 of pregnancy did not show a differential effect of d-A. It is concluded that early prenatal exposure to low doses of MM can result in behavioral consequences subtle enough to require unmasking of the effects with psychotropic drugs. Additionally, periods may exist during development when the embryo or fetus is most susceptible to behavioral or functional teratogenic effects of exposure to chemical insult. The testing procedures used in these experiments were objective, automatic and amenable for use with relatively large sample sizes compared with other operant behavior analytical methods. They also lend themselves to appropriate parametric statistical analyses, a staunch requirement for behavioral toxicological and teratological studies.     

Reduction of arginase I activity and manganese levels in the liver during exposure of rats to methylmercury: a possible mechanism

The toxicity of methylmercury (MeHg) is, in part, thought to be due to its interaction with thiol groups in a variety of enzymes, but the molecular targets of MeHg are poorly understood. Arginase I, an abundant manganese (Mn)-binding protein in the liver, requires Mn as an essential element to exhibit maximal enzyme activity. In the present study, we examined the effect of MeHg on hepatic arginase I in vivo and in vitro. Subcutaneous administration of MeHg (10 mg/kg) for 8 days to rats resulted in marked suppression of arginase I activity. With purified arginase I, we found that interaction of MeHg with arginase I caused the aggregation of arginase I as evaluated by centrifugation and subsequent precipitation, and then the reduction of catalytic activity. Experiments with organomercury column confirmed that arginase I has reactive thiols that are covalently bound to organomercury. While MeHg inhibited arginase I activity, Mn ions were released from this enzyme. These results suggest that MeHg-mediated suppression of hepatic arginase I activity in vivo is, at least in part, attributable to covalent modification of MeHg or substantial leakage of Mn ions from the active site.     

Prenatal exposure to lead and cognitive deficit in 7- and 14-year-old children in the presence of concomitant exposure to similar molar concentration of methylmercury

Few studies have examined the effects of mixed metal exposures in humans. We have evaluated the effect of prenatal lead exposure in a Faroese birth cohort in the presence of similar molar-level exposure to methylmercury. A cohort of 1022 singleton births was assembled in the Faroe Islands during 1986–1987 from whom lead was measured in cord-blood. A total of 896 cohort subjects participated in a clinical examination at age 7 and 808 subjects in a second examination at age 14. We evaluated the association between cord-blood lead concentrations and cognitive deficits (attention/working memory, language, visuospatial, and memory) using multiple regression models. Overall, the lead concentration showed no clear pattern of association. However, in subjects with a low methylmercury exposure, after inclusion of statistical interaction terms, lead-associated adverse effects on cognitive functions were observed. In particular, higher cord-blood lead was associated with a lower digit span forward score on the Wechsler Intelligence Scale for Children-Revised (WISC-R) [beta = ?1.70, 95% confidence interval (CI): ?3.12 to ?0.28] at age 7 and a lower digit span backward score on the WISC-R (beta = ?2.73, 95%CI: ?4.32 to ?1.14) at age 14. Some interaction terms between lead and methylmercury suggested that the combined effect of the exposures was less than additive. The present study indicates that adverse effects of exposure may be overlooked if the effects of a co-pollutant are ignored. The present study supports the existence of adverse effects on cognitive functions at prenatal lead exposures corresponding to an average cord-blood concentration of 16 μg/L.