Interaction of methylmercury and selenium on the bioaccumulation and histopathology in medaka (Oryzias latipes)

Interaction of methylmercury and selenium in medaka (Oryzias latipes) on bioaccumulation of pollutants and histopathological changes in liver and gill were studied. Juvenile medaka fish were submitted to a series of waterborne methylmercury chloride (MMC), sodium selenite (Na(2)SeO(3)) and their mixture for 210 days, respectively. The methylmercury (MeHg) and selenium contents in the whole body of medaka were determined. The dose- and time-dependent increase of MeHg and selenium contents in medaka were observed. Histopathological changes, such as edema, vacuoles, pyknotic nucleus, and telangiectasis, could clearly be observed in the slices from the exposed medaka's liver and gill. Concurrent exposure to MMC and Na(2)SeO(3) showed the increased selenium accumulation. When the exposure molar ratio of MeHg:Se was about 1, the interaction between MeHg and selenium offered a limited protection against the serious intoxication of both MMC and Na(2)SeO(3) to medaka.     

Effect of cystine,selenium, and fish protein on the toxicity and metabolism of methylmercury in rats

Investigations were made of the effect of 0.4% cystine, 0.6 ppm selenium, and fish protein on the toxicity and metabolism of mercury from methylmercuric chloride in male and female weanling rats. When diets with 10% protein and adequate amounts of other nutrients contained 25 ppm mercury, cystine reduced toxicity, selenium as sodium selenite had a greater effect in reducing toxicity, and the combination of cystine and selenium produced a considerably greater additive effect, as measured by increased growth and survival time. Toxicity signs were reduced when fish protein replaced casein in the basal diet, and a 20% protein level from either source reduced toxicity symptoms as compared to a 10% protein level. Cystine, selenium, and fish protein did not inhibit mercury toxicity by increasing its elimination from the body via the urine and feces and, in fact, slightly more mercury was retained when cystine and selenium were added to diets. Few differences were found in the deposition of mercury in tissues that could be attributed to cystine, selenium, and fish protein. The major difference found was in one experiment in which mercury concentrations in the kidney were considerably lower when diets contained cystine, selenium, and fish protein. Possible explanations for the effects observed in these studies included a reduction in mercury accumulation and, hence, damage in the kidney, complexing of mercury by the additives evaluated, and conversion of organic mercury to the less toxic inorganic form.     

Formation and possible role of bis(methylmercuric) selenide in rats treated with methylmercury and selenite

The metabolic fate of methylmercury after administration of [²⁰³Hg]-methylmercuric chloride in combination with sodium selenite was investigated in rats. Whole body autoradiography and radioassay showed that administration of selenite decreased the mercury concentration in the liver and kidney, and increased that in the brain. The rapid changes of methylmercury concentration in the tissues after selenite injection were accompanied by increases in mercury extractable with benzene at neutral pH. The maximum levels of benzene-extractable mercury in the blood, kidney and liver were attained 30 min after selenite injection and were 30, 23 and 8 percent, respectively, of the total mercury. Thin-layer chromatography showed that the benzene-extractable mercury was a complex of methylmercury with selenium, bis(methylmercuric) selenide. These findings indicate that selenite alters the distribution of methylmercury in the tissues by formation of a diffusible complex with methylmercury, bis(methylmercuric) selenide.     

Maternal selenium deficiency enhances the fetolethal toxicity of methyl mercury

The effect of maternal selenium deficiency on methyl mercury fetotoxicity was examined in the ICR strain of mice. Pregnant mice were fed either selenium-deficient diets based on torula yeast or selenium-supplemented diets which were identical to the former except that 0.1, 0.2, or 0.4 mg of selenium per kilogram of diet was added as sodium selenite. Fetolethality of methyl mercury was exacerbated by maternal selenium deficiency when mothers were administered sc 15, 25, or 35 mumol/kg/day of methylmercuric chloride (MMC) on the 13, 14, and 15th days of pregnancy. One-tenth part per million of selenium in the diet was sufficient to protect the fetuses against MMC fetolethality when dams were administered 25 mumol/kg/day of MMC. Mercury concentrations in maternal and fetal tissues were independent of the dietary selenium level. Selenium concentration and glutathione peroxidase (GSH-Px) activity in maternal tissues were unaffected by MMC administration. In fetal liver, on the other hand, selenium concentration was increased and GSH-Px activity was decreased concurrently by maternal MMC administration in the selenium-supplemented groups. Therefore, as far as GSH-Px activity was concerned, the bioavailability of selenium was markedly decreased in fetal liver by maternal injection of MMC. The increase in selenium content in fetal liver, which was observed only in the selenium-supplemented groups, may play an important role in protection against fetolethal toxicity of MMC.     

Effect of interaction between 65Zn,mercury and selenium in rats (retention,metallothionein, endogenous copper)

Interaction of zinc with mercuric chloride and sodium selenite was studied in the rat at the organ and subcellular levels (liver and kidneys). Zinc chloride was administered subcutaneously at dose of 5 mg Zn/kg, mercury chloride into the tail vein at a dose of 0.5 mg Hg/kg (both metals every other day during 2 weeks) and sodium selenite intragastrically, at doses of 0.1 mg Se/kg, every day. Zinc retention in the rat did not exceed 20% and was unchanged in the presence of mercury. An interaction effect was reflected by an increased whole-body retention of zinc by selenium, mercury, and selenium. In the presence of selenium no peak of metallothionein-like proteins stimulated by zinc or mercury was found in the soluble fraction of the kidneys. The metallothionein level did not differ from that typical for control group animals, too. A significant increase in the level of endogenous copper was found only in the kidneys of rats exposed to zinc in the presence of mercury and selenium.     

Acute toxicity of inorganic selenium to Daphnia magna (Straus) and the effect of sub-acute exposure upon growth and reproduction

The acute toxicities of sodium selenite (Na₂SeO₃) and sodium selenate (Na₂SeO₄) to Daphnia magna were determined in defined culture at 22°C. For adults, the 48-h LC₅₀ values were 0.68 ppm selenium as selenite and 0.75 ppm selenium as selenate. Juveniles were more sensitive, with a 48-h LC₅₀ of 0.55 ppm selenium as selenate. Eggs and embryos were found to be much less sensitive, with a 72-h LC₅₀ of 1.4 ppm selenium as selenate.

Sub-acute exposure of D. magna to sodium selenate caused suppression of growth over instars 1–5 and reduced egg production in instar 9 when adults were exposed to test solutions from instar 6 onwards. These sublethal effects were found at concentrations in the range proposed as suitable for the use of selenium in the amelioration of mercury contamination.     

Effects of dietary lipids on whole-body retention and organ distribution of organic and inorganic mercury in mice.

The effect has been investigated of dietary lipids on the whole-body retention and organ distribution of organic and inorganic mercury in mice. A single oral dose of methylmercury chloride or mercuric chloride labelled with 203Hg was given to female NMRI mice fed semi-synthetic diets containing varying amounts (5, 10, 20 or 50%) of energy derived from lipid (coconut oil, soya oil, or cod liver oil). The whole-body retention and relative organ distribution of mercury depended on diet composition. Thus, a significant reduction of the whole-body retention of mercury was seen in mice fed a diet containing 50% cod liver oil compared with mice fed a diet containing 50% coconut oil. After oral administration of mercuric chloride the relative deposition of mercury in the kidneys increased while that in the liver decreased with increasing concentrations of soya oil or coconut oil in the diet. The whole-body retention of mercury after treatment with methylmercury chloride was significantly decreased in mice fed cod liver oil compared with mice fed coconut oil; there was no difference between mice fed cod liver oil and those fed soya oil. The relative disposition of mercury was significantly higher in all organs of mice fed a diet containing 20% energy from cod liver oil compared with mice fed a diet containing 20% energy from soya oil. The present study demonstrates that diet composition is of major importance to the toxicokinetics of methylmercury and mercuric mercury.     

Simultaneous administration of sodium selenite and mercuric chloride decreases efficacy of DMSA and DMPS in mercury elimination in rats

Two chelating agents meso-2,3-dimercaptosuccinic acid (DMSA) and sodium 2,3-dimercapto-propane-1-sulphonate (DMPS) were tested for their efficiency in mercury removal from the body of rats in the presence and in the absence of selenium. Female Wistar rats were given a single intraperitoneal injection of mercuric chloride or an equimolar mixture of mercuric chloride and sodium selenite (1.5 micromol/kg body weight). The chelating agents were given orally, in excess (500 micromol DMSA/kg body weight; 300 micromol DMPS/kg body weight), 30 min after the administration of mercury and selenium. The animals were euthanized 24 h after the treatment and mercury in the kidney, liver, and 24 h urine was determined using cold vapour atomic absorption spectrometry (CV-AAS). The simultaneous administration of mercuric chloride and sodium selenite led to a redistribution of mercury in the organs, so that accumulation of mercury in the kidneys was decreased and in the liver increased. Selenite also caused decrease in the level of urinary mercury excretion. Both chelating agents were effective in mercury removal from the body, by increasing its urinary excretion. However, when animals were simultaneously treated with mercury and selenite, the rise of mercury excreted in the urine due to the treatment with chelating agents was lower when compared to animals receiving mercury without selenite. It is concluded that sodium selenite decreases the efficiency of DMSA and DMPS in mercury removal from the body of rats.     

硒对氯化汞免疫毒性的影响及其机理

采用免疫毒理学和生化毒理学方法，研究了亚硒酸钠对氯化汞免疫毒性的影响及作用机理。结果表明：氯化汞染毒的ＩＣＲ小鼠血中碳廓清率、ＳＲＢＣ致敏小鼠的ＤＴＨ反应、ＤＮＣＢ所致的ＤＣＨ反应、血清溶血素形成和免疫器官脏体比均明显低于对照组，预先投以亚硒酸钠后，再给同剂量的氯化汞的小鼠上述各指标都有不同程度提高。免疫器官脂质过氧化作用，汞组与对照组比较：脂质过氧化物含量明显升高，而谷胱甘肽过氧化物酶活性明显降低；硒汞组与汞组比较：脂质过氧化物含量明显降低，而谷胱甘肽过氧化物酶活性明显升高。上述结果提示：亚硒酸钠对氯化汞所致的免疫毒性和免疫器官的脂质过氧化损伤具有保护作用     

Whole-body retention of mercury and selenium and histopathological and morphological studies of kidneys and liver of rats exposed repeatedly to mercuric chloride and sodium selenite

Distribution and retention of mercury and selenium was studied in rats exposed repeatedly to HgCl₂ injections (0.5 mg Hg/kg to the tail vein every other day) and intragastrically to Na₂SeO₃ (0.5 mg Se/kg every day), applying combined and separate administration of these metals for 2 weeks. Whole-body retention of mercury in the presence of selenium was augmented by 20% and that of selenium in the presence of mercury by 4% with respect to the administered dose. Combined administration of mercuric chloride and sodium selenite brought about damage to the epithelial cells of renal proximal convolutions and formation of protein casts in their lumen. These changes had the same pattern as those induced by administration of mercuric chloride alone, but the intensity was lower. Submicroscopic studies revealed that repeated combined administration of sodium selenite and mercuric chloride did not completely abolish the mercury-induced mitochondrial swelling and contributed to chromatin destruction in the hepatocyte nuclei.This work was supported by the Section of Medical Sciences of the Polish Academy of Sciences (Agreement 537/VI)     

Hepatic glutathione metabolism and lipid peroxidation in response to excess dietary selenomethionine and selenite in mallard ducklings

Studies were conducted with mallard (Anas platyrhynchos) ducklings to determine the effects of excess dietary selenium (Se) on hepatic glutathione concentration and associated enzymes, and lipid peroxidation. Day-old ducklings were fed 0.1, 10, 20, or 40 ppm Se as seleno-DL-methionine or sodium selenite for 6 wk. Selenium from selenomethionine accumulated in a dose-dependent manner in the liver, resulting in a decrease in the concentration of hepatic-reduced glutathione (GSH) and total hepatic thiols (SH). These effects were accompanied by a dose-dependent increase in the ratio of oxidized glutathione (GSSG) to GSH, and an increase in malondialdehyde concentration as evidence of lipid peroxidation. Hepatic and plasma GSH peroxidase activity was initially elevated at 10 ppm Se as selenomethionine, whereas GSSG reductase activity was elevated at higher dietary concentrations of Se. Selenium from sodium selenite accumulated in the liver to an apparent maximum at 10 ppm in the diet, resulting in an increase in hepatic GSH and GSSG accompanied by a small decrease in total hepatic SH. Sodium selenite resulted in an increase in hepatic GSSG reductase activity at 10 ppm and in plasma GSSG reductase activity at 40 ppm. A small increase in lipid peroxidation occurred at 40 ppm. These findings indicate that excess dietary Se as selenomethionine has a more pronounced effect on hepatic glutathione metabolism and lipid peroxidation in ducklings than does selenite, which may be related to the pattern of accumulation. Effects of Se as selenite appear to be less pronounced in ducklings than reported in laboratory rodents. The effects of selenomethionine, which occurs in vegetation, are of particular interest with respect to the health of wild aquatic birds in seleniferous locations.     

Effects of methylmercury chloride on various cholinergic parameters in vitro

The effects of methylmercury chloride and other mercury compounds on cholinergic parameters were studied in vitro. Methylmercury chloride (MMC) and phenylmercury acetate inhibited choline acetyltransferase (ChA) with 20 microM of I50, and mercury nitrate (MN) with 100 microM of I50. All the three compounds had little effect on cholinesterase activity. MMC inhibited a high affinity choline uptake with 41 microM of Ki, as well as a low affinity choline uptake with 250 microM of Ki. MMC did not affect a spontaneous and potassium-stimulated ACh release from brain tissue slices incubated in eserinized Krebs-Ringer's solution up to the concentration of 100 microM. It was shown that the organic mercury compounds, such as methylmercury, were potent inhibitors of the choline uptake systems, as well as ChA activity.     

Involvement of tissue sulfhydryls in the formation of a complex of methylmercury with selenium

Albumin-bound methylmercury was converted to a benzene-extractable form by the soluble fraction of rat liver, kidney or brain in the presence of selenite, but not in its absence. The factors in the soluble fraction causing this conversion were investigated by column chromatography. Sephadex G-25 chromatography showed that effective factors were present in non-protein and protein fractions. It was concluded from ion exchange and Sephadex G-200 chromatography that these factors in the non-protein and protein fractions were reduced glutathione (GSH) and protein sulfhydryl groups respectively. Because GSH and the soluble protein could be replaced by sulfhydryl compounds, such as cysteine and 2-mercaptoethanol, as well as by a purified protein with sulfhydryl groups, reduced ribonuclease (RNase), respectively, it was concluded that sulfhydryl groups of GSH and/or proteins in the soluble fraction were needed for selenite-induced conversion of methylmercury to a benzene-soluble form. Among the various selenium compounds tested, only H₂Se (the reduced metabolite of selenite) was found to react directly with methylmercury to form a benzene extractable mercury compound in the absence of the soluble fraction. These findings suggest that the conversion of methylmercury to a benzene-soluble form occurs by reaction of methylmercury with selenium (possibly H₂Se) reduced by GSH and/or protein sulfhydryl groups in the soluble fraction. Thin-layer chromatography showed that benzene-extractable mercury consists mainly of bis(methylmercuric) selenide (BMS). A minor component, trismethylmercuric selenonium, was also detected by mass spectrography.     

Effect of selenium on the uptake of methyl mercury across perfused gills of rainbow trout Oncorhynchus mykiss

The uptake of methyl mercury was measured across the perfused gills of rainbow trout Oncorhynchus mykiss. The effect of selenium, either in the blood (perfusion medium), or in the water was investigated. Methyl mercury was effectively taken up from the water across the gills into the perfusate. The uptake rate reached a stable level after 30 min perfusion. When the gills were placed in mercury free water after exposure to mercury in the water for 1 h, they continued to liberate significant amounts of accumulated mercury into the perfusate. Exposure to selenite (SeIV) or selenate (VI) (0.075–0.75 μM) in the external medium did not affect the uptake of methyl mercury across the gills or the liberation of the metal from the gills. Internal selenite or selenate (7.5 μM) augmented the uptake of methyl mercury across the gills and internal selenite also increased the amounts of liberated methyl mercury from the gills in the unload period. Internal selenium, increased the mercury accumulation in the gills, whereas, external selenium did not alter the mercury accumulation in the gills. Uptake of selenium from the water across the gills occurred very slowly.     

Methylmercury stimulates the exhalation of volatile selenium and potentiates the toxicity of selenite

The aim of the present experiments was to investigate whether a single dose of 24 mumol/kg methylmercuric chloride (MeHgCl) in rats can influence the effect of an equimolar dose of sodium selenite (Na2SeO3) on body weight or the exhalation of dimethylselenide, a volatile metabolic product of selenium. Due to the difference in their single-dose toxicities, only selenite depressed body weight gain, when given alone. The experiments indicated that methylmercury, irrespective of whether it was given 1-2 h before, or at the same time as sodium selenite, potentiated the effect of the latter on body weight. Methylmercury also increased the exhalation of volatile selenium, but this effect decreased when the administration of selenite was delayed.     

Short-term (6-week) oral toxicity study of selenium in Syrian hamsters

The subacute toxicity of selenium was tested by feeding sodium selenite to Syrian hamsters at dietary levels of 0.1, 1, 5, 10 and 20 ppm selenium for 42 days. General health, survival and organ weights were not adversely affected at any of the dose levels. Weight gain and food intake were relatively low in males fed the highest dose level. The differences from the control values were not statistically significant and there was no similar effect in females. Hamsters fed 10 or 20 ppm retained considerably higher levels of selenium in the liver than did the controls. Microscopic examination of the liver revealed degenerative changes in males and females in the 20-ppm group. The no-toxic-effect level of selenium fed in the diet for 42 days to Syrian hamsters was found to be 10 ppm, equivalent to an intake of about 0.7 mg selenium/kg body weight/day.     

Effects of sodium selenite on methylmercury distribution in mice of late gestational period

Mercury distribution in pregnant mice, subcutaneously injected with 1.5 or 15.0 mol/kg of methylmercury chloride (MeHg) and 0, 1.5 or 15.0 mol/kg of sodium selenite, was investigated.Selenite increased the retention of mercury in maternal brain in every combination of doses. Selenite also increased mercury concentrations in maternal blood except one combination (MeHg 1.5; selenite 1.5 mol/kg), and the increased mercury was partitioned to red blood cells. The increased mercury retention by selenite was also found in fetal brain.     

Embryotoxic and teratogenic effects of selenium in the diet of mallards

Mallards (Anas platyrhynchos) were fed a control diet, diets containing 1, 5, 10, or 25 ppm Se as sodium selenite, or a diet containing 10 ppm Se as seleno-DL-methionine in the first of two experiments. Selenium at 10 ppm as selenomethionine or 25 ppm as sodium selenite caused a 40-44% decrease in the total number of eggs that hatched compared to controls. Selenium at 25 ppm (sodium selenite) resulted in a 19% decrease in mean embryonic weight at 18 d of incubation, accompanied by a 6% decrease in crown-rump length. Ten parts per million Se as selenomethionine was more teratogenic than sodium selenite at 25 ppm. Selenomethionine (10 ppm Se) resulted in an incidence of 13.1% malformations that were often multiple, whereas sodium selenite (10 and 25 ppm Se) resulted in 3.6 and 4.2% malformations. The teratogenicity of selenomethionine was confirmed in a second experiment in which mallards received 1, 2, 4, 8, or 16 ppm Se as selenomethionine, resulting in 0.9, 0.5, 1.4, 6.8, and 67.9% malformations, respectively. These malformations included hydrocephaly, microphthalmia, lower bill defects, and foot defects with ectrodactyly. Both forms of selenium increased the incidence of edema and stunted embryonic growth. Selenomethionine (10 ppm Se) resulted in a significant increase of approximately 40% in plasma glutathione peroxidase activity and a 70% increase in sorbitol dehydrogenase activity (indicative of hepatotoxicity) in hatchlings. Sodium selenite (25 ppm Se) resulted in fourfold elevation in plasma uric acid concentration, indicative of renal alteration. Selenomethionine accumulated much better in eggs than did sodium selenite. These findings indicate that selenomethionine is considerably more teratogenic and generally more embryotoxic than sodium selenite, probably due to higher uptake of selenomethionine.     

Prenatal methylmercury exposure increases responding under clocked and unclocked fixed interval schedules of reinforcement

Recent experiments have suggested that developmental methylmercury exposure produces perseverative behavior in adulthood. In the present experiment, interactions between developmental low-level methylmercury (MeHg) and nutritionally relevant dietary selenium (Se) on operant behavior and its persistence were examined in aged animals. Female rats were exposed, in utero, to 0, 0.5, or 5 ppm mercury as MeHg via drinking water, approximating mercury exposures of 0, 40, and 400 microg/kg/day. They also received both pre- and chronic post-natal exposure to a diet that was marginal (0.06 ppm) or rich (0.6 ppm) in Se, a nutrient believed to protect against MeHg's toxicity. This created a 2 (chronic Se)x3 (gestational MeHg) full factorial design, with 6-8 female rats per cell. At eleven months of age, a multiple schedule consisting of alternating fixed interval (FI) and clocked FI (CFI) components was arranged. The CFI component was divided into 5, 24-second bins, each associated with a different auditory stimulus, providing a "clock." Low and high response rates were evaluated using the initial 40% (bins 1 and 2) and last 20% (bin 5) of the FI and CFI components, respectively. Rats exposed to 5 ppm Hg made more responses than the other two groups during the last 20% of the intervals, regardless of selenium exposure or presence of the clock stimuli. They did not differ from the other groups during the initial 40% of the FI and CFI components. Following reinforcement omission for half of the intervals at 21 months of age, the 5 ppm Hg group continued to respond at higher rates than the other groups in both components.     

Effect of various amounts of selenium on the metabolism of mercuric chloride in mice

Male ddY mice were given one injection of (1) mercury (mercuric chloride) simultaneously with various doses of selenium (sodium selenite), (2) mercury alone, or (3) various doses of selenium alone. The interaction between mercury and selenium in the liver and kidneys at 1, 5, 24, 120, and 240 hr after administration was investigated. The concentrations of mercury in the liver of mice receiving mercury and selenium simultaneously were higher than those after administration of mercury alone, while the concentrations of mercury in the kidney decreased markedly over a 1-120 hr period after administration, depending on the dose of selenium administered simultaneously with mercury. Clearly, selenium had a different effect on the accumulation of mercury in the liver and kidneys. Subcellular distribution studies revealed that mercury and selenium which were administered simultaneously were incorporated into the crude nuclear and mitochondrial fractions as stable complexes. The transport of these complexes to the kidneys seems to be limited. In addition, gel filtration of supernatant fractions of liver and kidney through a Sephadex G-75 column indicated that the proportion of mercury bound to metallothionein fraction decreased depending on the dose of selenium administered simultaneously with the mercury. This reduction was attributed to the decreased synthesis of mercury-thionein due to a reduction in the activity of Hg2+ which results from binding between mercury and selenium in the cells.