The biological monitoring of prenatal exposure to methylmercury.

Several biological media have been used as indicators of the fetal body burden of methylmercury and the levels in the primary target tissue, the developing brain. These media include maternal hair and blood. The relative merits of these media will be considered both with regard to current knowledge of the physiology of mercury disposition in the body and also the practicality of field application with respect to sample, collection, transport, storage and processing.

Perinatal and lifetime exposure to methylmercury in the mouse: blood and brain concentrations of mercury to 26 months of age.

Chronic, low-level exposures to environmental toxicants, because they often begin prenatally and then persist throughout the individual's lifetime, pose challenging issues to risk assessment. Exposure to low levels of methylmercury through the diet, based largely on consumption of fish and sea mammals, follows this pattern. Early development is considered to be a period of heightened vulnerability during which even low-level exposures may produce undetected, "silent", damage that is revealed only under conditions that challenge the functional capacities of the individual. Aging, with its diminished functional capacities and compensatory reserves provides such a challenge, but, to explore this possibility, requires basic information about blood and brain levels under conditions of chronic lifetime exposure. The current research was undertaken to provide such information. One hundred female B6C3F1/HSD mice were assigned to one of three dose groups, 0, 1, or 3 ppm methylmercury chloride administered in a 5 nM sodium carbonate drinking solution. They were bred with male CBA/J HSD mice to produce the trihybrid offspring B6C3F1/ HSD x CBA/J HSD. Dosing of the females began 4 weeks prior to breeding and continued for the two methylmercury-exposed groups throughout breeding and gestation. The methylmercury-treated litters were split into two subgroups, one exposed throughout its lifetime (set at 26 months) to the original dose, the other exposed through postnatal day 13 (PND 13). Brain and blood concentrations were assayed by cold-vapor atomic absorption. Samples were obtained on PND 4 and 21, and then at the end of months 14 and 26. On PND 4, brain and blood levels closely reflected maternal dosing. In all groups, concentrations fell sharply from PND 4 to 21, but to a greater extent in the perinatal groups. Blood levels in the 1 ppm lifetime group remained unchanged between months 14 and 26, but brain levels rose modestly. In the 3 ppm lifetime group, both brain and blood levels rose significantly between months 14 and 26, suggesting an interaction between dose and age.

Association between methylmercury exposure from fish consumption and child development at five and a half years of age in the Seychelles Child Development Study: an evaluation of nonlinear relationships.

Studies to date of the developmental effects of pre- and postnatal methylmercury exposure from fish consumption in the Seychelles Islands, using linear regression models for analysis, have not shown adverse effects on neurodevelopmental test scores. In this study we evaluated whether nonlinear effects of methylmercury exposure were present, using scores on six tests administered to cohort children in the Seychelles Child Development Study at 66 months of age. Prenatal exposure was determined by measuring mercury in a segment of maternal scalp hair representing growth during pregnancy. Postnatal exposure was measured in a segment of the child's hair taken at 66-months of age. Generalized additive models (GAMs), which make no assumptions about the functional form of the relationship between exposure and test score, were used in the analysis. GAMs similar to the original linear regression models were used to reanalyze the six primary developmental endpoints from the 66-month test battery. Small nonlinearities were identified in the relationships between prenatal exposure and the Preschool Language Scale (PLS) Total score and Child Behavior Check List (CBCL) and between postnatal exposure and the McCarthy General Cognitive Index (GCI) test scores. The effects are best described graphically but can be summarized by computing the change in the predicted test score from 0 to either 10 or 15 ppm and then above this point. For the PLS the trend involved a decline of 0.8 points between 0 and 10 ppm followed by an increase (representing improvement) of 1.3 points above 10 ppm. For the CBCL there was an increase of 1 point from 0 to 15 ppm, and then a decline (improvement) of 4 points above 15 ppm. The GCI increased by 1.8 points through 10 ppm and then declined 3.2 points (representing worse performance) above 10 ppm. These results are not entirely consistent. Two of the trends involve what appear to be beneficial effects of prenatal exposure. The one possibly adverse trend involves postnatal exposure. In every case the trend changes direction, so that an effect in one direction is followed by an effect in the opposite direction. Because of the descriptive nature of GAMs it is difficult to provide a precise level of statistical significance for the estimated trends. Certainly above 10 ppm there is less data and trends above this level are estimated less precisely. Overall there was no clear evidence for consistent (across the entire range of exposure levels) adverse effects of exposure on the six developmental outcomes. Further nonlinear modeling of these data may be appropriate, but there is also the risk of fitting complex models without a clear biological rationale.

Association between prenatal exposure to methylmercury and cognitive functioning in Seychellois children: a reanalysis of the McCarthy Scales of Children's Ability from the main cohort study.

Methylmercury (MeHg) is a neurotoxicant whose high-dose effects first became known following a number of poisoning outbreaks that occurred worldwide. The primary human exposure is low dosage from fish consumption. Studies of fish-eating populations have not found a consistent pattern of association between exposures and outcomes. Therefore, examining specific areas of cognitive functioning has been suggested as an important approach to determine whether more subtle effects of MeHg exposure are present. In the Seychelles longitudinal study of prenatal and postnatal MeHg exposure from fish consumption and development, the McCarthy Scales of Children's Abilities (MSCA) were administered to children at age 66 months. No association between MeHg exposure and performance on the MSCA General Cognitive Index was identified. We analyzed these data further to determine whether associations were present on specific subscales of the MSCA. The standard MSCA subscales were analyzed. Then, more specific subscales of the MSCA were defined and analyzed utilizing a neuropsychological approach. The subscales were recombined to approximate the domains of cognitive functioning evaluated in the Faroes and New Zealand studies. Analyses of both the standard and the recombined MSCA subscales showed no adverse associations with MeHg exposure and neuropsychological endpoints. A positive association between postnatal MeHg exposure and performance on the MSCA Memory subscale was found. These findings are consistent with previous reports from the Seychelles study in that no adverse effects of MeHg exposure from fish consumption can be detected in this cohort.

Segmental hair mercury evaluation of a single family along the upper Madeira basin, Brazilian Amazon.

Mercury pollution (MeHg) up the aquatic food chains in the Amazonian ecosystems has been a major concern in environmental health. Riverside people (ribeirinhos) along the Upper Madeira river are heavy fish eaters. Hair is the best biomarker for MeHg exposure. By assuming a constant hair growth rate, it is possible to evaluate a temporal profile of Hg exposure over the recent defined past. In this paper we present the segmental total hair Hg concentrations from a single family from which some of the 10 persons investigated had high hair Hg concentrations (peak of 339 ppm). We also presented the hair MeHg content from 4 out of the 10 family members investigated. There was a wide variation in total hair Hg concentrations (8 to 339 ppm) among these individuals, who were mostly sharing their meals; there was also a wide variation in total Hg concentrations in the same individual over time (136 to 274 ppm). Hg speciation showed a mean and standard deviation in the MeHg content of 62% and 6%, respectively. The wide variation in total hair Hg concentration strongly indicated that it is possible to mitigate critical Hg exposure levels by conducting a fish advisory.

Neurodevelopmental outcomes of Seychellois children from the pilot cohort at 108 months following prenatal exposure to methylmercury from a maternal fish diet.

The Seychelles Child Development Study has been unable to confirm any relationship between maternal exposure to MeHg during pregnancy and adverse developmental outcomes. In this report, 87 children from a pilot cohort were evaluated at 9 years of age. Each child was given a battery testing specific cognitive, visual motor, and motor skills using standardized psychometric and neuro-psychological tests. The results indicated no adverse association between maternal MeHg exposure and any developmental outcome measure. For three endpoints (Boston Naming Test and two tests of visual motor coordination), enhanced performance in males was associated with increasing prenatal MeHg exposure. A secondary analysis including both prenatal MeHg and postnatal MeHg exposure was done even though we lacked postnatal hair for about 35% of the cohort. The results of the secondary analysis mirrored the outcomes of the primary analysis regarding prenatal exposure but were less robust. The results of this study are consistent with earlier findings from the 66-month evaluations of the SCDS Main cohort. Since MeHg is neurotoxic, this effect is likely due to other factors associated with consumption of fish.

Secondary analysis from the Seychelles Child Development Study: the child behavior checklist.

Human exposure to methylmercury (MeHg), a known neurotoxin, is primarily from fish consumption. As part of a large study examining the association between MeHg exposure and child development in a population with high fish consumption we examined school-age behavior using the Achenbach Child Behavior Checklist (CBCL). The CBCL Total T score was a primary endpoint and was reported earlier to show no adverse association with prenatal or postnatal MeHg exposure. In this study we analyzed the T scores of the CBCL subscales to determine if more discrete aspects of measured behavior were associated with exposure. The Seychelles Child Development Study (SCDS) is a prospective, double blind, longitudinal evaluation of over 700 children. The index of prenatal exposure was maternal hair total mercury (T-Hg) in a segment growing during gestation. Postnatal exposure was T-Hg in the child's hair taken at 66 months of age. The child's primary caregiver completed the CBCL during the 66-month evaluation. No association between prenatal or postnatal MeHg exposure and the CBCL subscales was found. In Seychellois children exposed to MeHg from consumption of ocean fish we found no association between either prenatal or postnatal MeHg exposure and behavior as measured by the CBCL subscales.

Determination of methyl mercury in whole blood by ethylation-GC-CVAFS after alkaline digestion-solvent extraction.

A method for the determination of methyl mercury in whole blood samples based on ethylation-gas chromatography-cold vapor atomic fluorescence spectrometry after alkaline digestion-solvent extraction is described. The extraction procedure and conditions were optimized, and the matrix interference after extraction was critically investigated. The storage stability of MeHg in blood samples and a series of extracts was determined. The method detection limit was found to be approximately 0.02 ng/g for a 0.5-g blood sample with relative standard deviations of less than 10%. The accuracy and precision were evaluated by summarizing the quality-control (QC) data generated over a one and one half year period. Appropriate procedures for sample collection, transportation, and storage were adapted to the method. Using this method accompanied by explicit QC protocols and procedures, background levels of MeHg and total mercury in blood for 150 8-10-year-old Portuguese children with nonoccupational and nonamalgamal exposure were determined and reported with summarized QC data.

Twenty-seven years studying the human neurotoxicity of methylmercury exposure.

Research at the University of Rochester (U of R) has been focused on mercury for nearly half a century. Initially studies focused on dosimetry, especially the accuracy of measuring exposure, and experimental work with animal models. Clinical studies in human populations started when the U of R mercury group was asked to assist with dosimetry in the Iraq epidemic of 1971-1972. Initial clinical studies described the effects of methylmercury (MeHg) poisoning on adults and children. A dose-response curve for prenatal exposure was determined and it suggested that relatively low exposures might be harmful to the fetus. Since most human exposure to MeHg is dietary from fish consumption, these theoretical dangers had far-reaching implications. After Iraq, the Rochester team pursued exposure from fish consumption in both adults and children. Populations with high fish consumption were identified in Samoa and Peru for studying adults and in Peru and the Seychelles islands for studying children. The possible health threat to the fetus from maternal fish consumption quickly became the focus of research efforts. This paper reviews the Rochester experience in studying human exposure to MeHg from fish consumption.

A cluster of pediatric metallic mercury exposure cases treated with meso-2,3-dimercaptosuccinic acid (DMSA)

Nine children and their mother were exposed to vapors of metallic mercury. The source of the exposure appears to have been a 6-oz vial of mercury taken from a neighbor's home. The neighbor reportedly operated a business preparing mercury-filled amulets for practitioners of the Afro-Caribbean religion Santeria. At diagnosis, urinary mercury levels in the children ranged from 61 to 1,213 microg/g creatinine, with a geometric mean of 214.3 microg/m creatinine. All of the children were asymptomatic. To prevent development of neurotoxicity, we treated the children with oral meso-2,3-dimercaptosuccinic acid (DMSA). During chelation, the geometric mean urine level rose initially by 268% to 573.2 microg mercury/g creatinine (p<0.0005). At the 6-week follow-up examination after treatment, the geometric mean urine mercury level had fallen to 102.1 microg/g creatinine, which was 17.8% of the geometric mean level observed during treatment (p<0.0005) and 47.6% of the original baseline level (p<0.001). Thus, oral chelation with DMSA produced a significant mercury diuresis in these children. We observed no adverse side effects of treatment. DMSA appears to be an effective and safe chelating agent for treatment of pediatric overexposure to metallic mercury.

Mercury contamination of heavy metal collection containers.

We investigated discordant urinary mercury testing results from 2 patients with potential mercury exposures. Two patients had mercury levels of 634 and > 1,000 micrograms/L respectively. Although repeat 24 h urine mercury levels were elevated, spot urines were negative. Investigation revealed that technical HCl with high mercury content had been added to the 24 h urine collection containers. Subsequently, 20 hospitals were contacted to determine their heavy metals testing procedure and to analyze the acid used for mercury. Most hospitals contacted used acid in the preparation of their urine heavy metal collection containers. Of 13 HCl samples tested, 5 had low levels of mercury and 1 had heavy mercury content. Acid added to heavy metal collection containers should be of high purity grade to avoid mercury contamination of samples.

Association between prenatal exposure to methylmercury and developmental outcomes in Seychellois children: effect modification by social and environmental factors.

UNLABELLED: The Seychelles Child Development Study (SCDS) is testing the hypothesis that prenatal exposure to low doses of MeHg from maternal consumption of fish is associated with the child's developmental outcomes. No deleterious relationships between exposure to MeHg and cognitive functions have been identified in the primary analysis of the main cohort through 66 months of age. We performed secondary analyses to determine if effect modification (EM) from social and environmental factors was affecting associations between MeHg and outcomes. METHODS: MeHg exposure was determined by analysis of maternal hair growing during pregnancy. Children in our Main Study cohort were evaluated at 6.5 months (N = 740) for visual recognition memory and visual attention using the Fagan Infantest, at 19 months (N = 738) and 29 months (N = 736) with the Bayley Scales of Infant Development (BSID). Interactions between MeHg and Caregiver Intelligence, Family Income and Home Environment were examined by multiple regression analysis. RESULTS: The median prenatal MeHg exposure was 5.9 ppm (Range 0.5-26.7 ppm). No EM occurred for preferential looking or visual attention at 6.5 months, for the BSID Psychomotor Scale at either 19 or 29 months, or for activity level at 29 months as measured by the BSID Infant Behavior Record. Interactions between MeHg level and both caregiver intelligence and family income were statistically significant for the BSID Mental Scale at 19 months but not at 29 months. These showed enhancement of MDI scores with increasing maternal MeHg in higher caregiver IQ groups at several levels of family income. CONCLUSIONS: In Seychellois children, consistent major EM by social or environmental factors were not identified. The small EM by caregiver intelligence and social factors at 19 months is consistent with the enhanced performance we reported when this cohort was examined at 66 months.

Effects of prenatal and postnatal methylmercury exposure from fish consumption on neurodevelopment: outcomes at 66 months of age in the Seychelles Child Development Study.

CONTEXT: Human neurodevelopmental consequences of exposure to methyl-mercury (MeHg) from eating fish remain a question of public health concern. OBJECTIVE: To study the association between MeHg exposure and the developmental outcomes of children in the Republic of Seychelles at 66 months of age. DESIGN: A prospective longitudinal cohort study. PARTICIPANTS: A total of 711 of 779 cohort mother-child pairs initially enrolled in the Seychelles Child Development Study in 1989. SETTING: The Republic of Seychelles, an archipelago in the Indian Ocean where 85% of the population consumes ocean fish daily. MAIN OUTCOME MEASURES: Prenatal and postnatal MeHg exposure and 6 age-appropriate neurodevelopmental tests: the McCarthy Scales of Children's Abilities, the Preschool Language Scale, the Woodcock-Johnson Applied Problems and Letter and Word Recognition Tests of Achievement, the Bender Gestalt test, and the Child Behavior Checklist. RESULTS: The mean maternal hair total mercury level was 6.8 ppm and the mean child hair total mercury level at age 66 months was 6.5 ppm. No adverse outcomes at 66 months were associated with either prenatal or postnatal MeHg exposure. CONCLUSION: In the population studied, consumption of a diet high in ocean fish appears to pose no threat to developmental outcomes through 66 months of age.

Semiparametric modeling of age at achieving developmental milestones after prenatal exposure to methylmercury in the Seychelles child development study.

Controversy exists concerning the fetal risk associated with exposure to low-dose methylmercury from maternal fish consumption. Previous studies of the effects of acute prenatal mercury exposure identified delays in achieving developmental milestones among exposed children. This led to public health concern that prenatal low-dose exposure from fish consumption could adversely affect the fetus. We evaluated the effects of prenatal methylmercury exposure (through maternal fish consumption) on the age that children walked and first said words in the main study cohort of the Seychelles Child Development Study. We used semiparametric generalized additive models to identify nonlinearities in the relationships between prenatal exposure and developmental outcomes, after adjusting for covariates, and to evaluate their importance. Very slight delays (<1 day) in walking were seen as mercury levels increased from 0 to 7 ppm, but this effect did not persist at the higher exposure levels represented by the cohort, making it difficult to conclude that a cause and effect relationship existed at the exposure levels seen in this cohort. There was no evidence for any association between prenatal exposure and age at talking.

Longitudinal hair mercury concentration in riverside mothers along the Upper Madeira river (Brazil).

Mercury releases from gold mining occurred during the 1980s in the Upper Madeira river, a Southern tributary of the Amazon. Downstream from these areas, riverside residents rely on fish consumption for subsistence. In July of 1993, hair samples were collected for mercury analysis from a group of mothers and their infants and one pregnant woman. By assuming a constant rate of hair growth (1.1 cm per month), a temporal profile of the methylmercury exposure was determined for the previous 2 to 3 years. The length of hair segments corresponded to hair growth during pregnancy and the subsequent breastfeeding periods. During all periods, hair mercury concentrations in six mothers were in the range of 4.5-26.8 ppm, slightly lower than those of their infants (8.2-28.4 ppm). Further segmental analyses of hair mercury from another six mothers showed concentrations in the range of 12.2-41.0 ppm during the three trimesters of pregnancy and 4.0-33.5 ppm during breast feeding-slightly lower than their infants (11.6-50.4 ppm). Another four mothers showed hair mercury concentrations in the range of 21. 3-84.4 ppm.

Effects of prenatal methylmercury exposure from a high fish diet on developmental milestones in the Seychelles Child Development Study.

Mercury is widespread in the environment and exists in several physical and chemical forms. Prenatal exposure to methylmercury disrupts brain development. The most common mode of prenatal methylmercury exposure is maternal fish consumption. Studies of human prenatal exposure in Iraq following maternal ingestion of methylmercury treated grain suggested that maternal hair mercury concentrations above 10 ppm may be related to delayed developmental milestones and neurological abnormalities. This level of exposure can be achieved by frequent consumption of fish. The Seychelles Child Development Study analyzed developmental milestones similar to those determined in Iraq in a large controlled, prospective study of children exposed prenatally to methylmercury when their mothers ate fish. As part of this ongoing study, cohort children were evaluated at 6.5, 19, 29, and 66 months of age. At 19 months care-givers were asked at what age the child walked (n=720 out of 738) and talked (n=680). Prenatal mercury exposure was determined by atomic absorption analysis of maternal hair segments corresponding to hair growth during the pregnancy. The median mercury level in maternal hair was 5.8 ppm with a range of 0.5-26.7 ppm. The mean age (in months) at walking was 10.7 (SD = 1.9) for females and 10.6 (SD = 2.0) for males. The mean age at talking (in months) was 10.5 (SD = 2.6) for females, and 11.0 (SD = 2.9) for males. After adjusting for covariates and statistical outliers, no association was found between the age at which Seychellois children walked or talked and prenatal exposure to mercury. Normal ages at achievement of the developmental milestones walking and talking were found in Seychellois toddlers following prenatal exposure to methylmercury from a maternal fish diet. These results do not support the lowest effect levels in young children following prenatal methylmercury exposure predicted by the dose response analysis of the Iraq data. More detailed studies in older children are needed to determine if there are adverse effects in fish eating populations.

Simple solvent extraction technique for elimination of matrix interferences in the determination of methylmercury in environmental and biological samples by ethylation-gas chromatography-cold vapor atomic fluorescence spectrometry.

A solvent extraction technique involving no critical clean-up steps was developed for the determination of methylmercury (MeHg) in environmental and biological samples by aqueous phase ethylation, room temperature precollection, gas chromatographic separation and cold vapor atomic fluorescence spectrometric detection. Samples were first digested with KOH-methanol. then acidified prior to extraction with methylene chloride. MeHg was back-extracted from the solvent phase into water prior to aqueous phase ethylation. Recoveries close to 100% were obtained with RSDs less than 5% for all samples analyzed, making direct standardization possible. The detection limits were about 0.08 ng g(-1) when analyzing 0.1 g of dry sea plant homogenate and 0.02 ng g when analyzing 0.5 g of wet sediment samples. Various certified reference materials and intercomparison samples, including sediments, sea plants and tissues, were analyzed, and the results were in good agreement with the certified values. The technique was applied to the determination of MeHg in both sea plants from the Atlantic and the red blood protein of dolphins from the Mediteranean Sea. in sediments from the Mediterranean Sea and Minnesota rivers and in soils from different origins. Concentrations of MeHg in dolphin red blood protein samples were as high as 300 ng g(-1).

Biological monitoring of iodine, a water disinfectant for long-term space missions.

In order to establish guidelines for exposure of astronauts to iodine, used as a water disinfectant in space, we studied the usefulness of hair, saliva, and urine for biological monitoring in humans and in the human hair/nude mouse model. The monitoring of iodine in patients that received 150 mCi of Na131I (carrier-free) showed similar patterns of elimination for blood, saliva, and urine. The mean correlation coefficient (r) between iodine elimination for blood/saliva was 0.99, for blood/urine, 0.95, and for saliva/urine, 0.97. The absolute value of iodine concentrations in urine revealed marked variability, which was corrected by adjusting for creatinine levels. The autoradiographic studies of human hair demonstrated that iodine is rapidly incorporated into external layers of the hair root and can be removed easily during washing. These data were confirmed after iodine exposure using the human hair/nude mouse model. Hair does not provide satisfactory information about exposure due to unstable incorporation of iodine. The most useful medium for biological monitoring of astronauts exposed to high doses of iodine in drinking water is urine, when adjusted for creatinine, and saliva, if quantitative evaluation of flow rate is provided.