Trophic transfer of citrate, PVP coated silver nanomaterials, and silver ions in a paddy microcosm.

We used replicated paddy microcosm systems to estimate the tropic transfer of citrate-coated silver nanoparticles (AgNP citrate), polyvinylpyrrolidone (PVP)-coated AgNP (AgNP PVP), and silver ions (AgNO3) for 14 days under two exposure regimes (a single high-dose exposure; 60 µg L-1 and a sequential low-dose exposure at 1 h, 4 days and 9 days; 20 µg L-1 × 3 = 60 µg L-1). Most Ag ions from AgNO3 had dispersed in the water and precipitated partly on the sediment, whereas the two Ag NPs rapidly coagulated and precipitated on the sediment. The bioconcentration factors (BCFs) of Ag from AgNPs and AgNO3 in Chinese muddy loaches and biofilms were higher than those of river snails in both exposure conditions. These BCFs were more prominent for 14 days exposure (7.30 for Chinese muddy loach; 4.48 for biofilm) in the low-dose group than in the single high-dose group. Their retention of AgNPs and Ag ions differed between the two exposure conditions, and uptake and elimination kinetics of Ag significantly differed between AgNP citrate and AgNP PVP in the sequential low-dose exposure. Stable isotopes analyses indicated that the trophic levels between Chinese muddy loaches and biofilms and between river snails and biofilms were 2.37 and 2.27, respectively. The biomagnification factors (BMFs) of AgNPs and AgNO3 between Chinese muddy loaches and biofilms were significantly higher than those between river snails and biofilms under both exposure settings. The BMFs of AgNP citrate and AgNO3 between Chinese muddy loaches and biofilms were greater than those of AgNP PVP for 14 days in the single high-dose group, whereas the BMFs of AgNP PVP were greater than those of AgNP citrate and AgNO3 in the sequential low-dose group. These microcosm data suggest that AgNPs have the potential to impact on ecological receptors and food chains.

Mitochondrial genome of the black-throated loon, Gavia arctica (Gaviiformes: Gaviidae): phylogeny and evolutionary history.

We sequenced the complete mitochondrial (mt) genome of Gavia arctica. The circular mt genome is 17,065 bp long, consisting of 37 genes (13 proteins, 22 transfer RNAs, and two ribosomal RNAs) and a control region. Phylogenetic analysis based on the full mt genome sequences confirmed that the genus Gavia is a monophyletic group, containing the G. stellata, G. arctica, and G. pacifica. These data can provide insights into the phylogenetic relationships for inferring the pattern and degree of mt genome evolution among the loon species.

Complete mitochondrial genome of the Grey-capped Greenfinch subspecies, Chloris sinica ussuriensis (Passeriformes: Fringillidae).

We sequenced the complete mitochondrial (mt) genome of Chloris sinica ussuriensis. The circular mt genome is 16,813 bp long and encodes 13 proteins, 22 transfer RNAs, and 2 ribosomal RNAs. Phylogenetic analysis based on full mt genome sequences confirmed that the C. s. ussuriensis is monophyletic group of the Chloris sinica. The complete mitochondrial genome of C. s. ussuriensis can provide a valuable data for resolving geographical distribution of evolutionary subdivision within the C. sinica species in East Asia.

Intraspecific variation and phylogeographic patterns of the grey-capped greenfinch Chloris sinica ssp. (Passeriformes: Fringillidae).

To study the intraspecific variation of the grey-capped greenfinch Chloris sinica (Passeriformes: Fringillidae), we sequenced complete mitochondrial (mt) genome of the C. sinica ssp. prevalent in Ulleung Island, Republic of Korea. The full length of the genome is 16,812 bp, containing 37 genes (2 rRNAs, 13 proteins, and 22 tRNAs) with a putative control region (D-loop). A total of 98 single nucleotide polymorphisms (SNPs) in the full mt genome were retained for Ulleung Island population and these SNPs were greater than those of inland population compared to the reference China subspecies. The analysis of the SNPs revealed 18 SNPs for ND4 gene, with a dominant haplotype shared by Ulleung population compared to the reference China population. The phylogenetic analysis of the C. sinica subspecies showed that they are monophyletic, however, there is clear phylogenetic separation between China and Korea subspecies with strong support (100% bootstrap). These data will provide new insights into reconstruct the intraspecific phylogeographical patterns of C. sinica species.

Complete mitochondrial genome of a treefrog, Hyla sp. (Anura: Hylidae).

The complete mitochondrial genome of a treefrog (Hyla sp.) was determined. The circular mitochondrial genome is 18,288 bp long and encodes 13 proteins, 22 transfer RNAs, and 2 ribosomal RNAs. Phylogenetic analysis of its full genome sequences showed that H. sp. was closely related to H. ussuriensis and H. japonica rather than Dryophytes suweonensis, consistent with the results from each protein-coding gene and a cluster between 12S rRNA and 16S rRNA. The present study will provide essential genomic information for biogeographical distribution and evolutionary history of an endemic treefrog, H.sp.

Influence of different types of nanomaterials on their bioaccumulation in a paddy microcosm: a comparison of TiO2 nanoparticles and nanotubes.

We investigated the environmental fate and bioaccumulation of TiO2 nanomaterials in a simplified paddy microcosm over a period of 17 days. Two types of TiO2 nanomaterials, nanoparticles (TiO2-NP) and nanotubes (TiO2-NT), were synthesized to have a negative surface charge. Ti concentrations in the environmental media (water, soil), crops (quillworts, water dropworts), and some lower and higher trophic organisms (biofilms, algae, plant-parasitic nematodes, white butterfly larva, mud snail, ricefish) were quantified after exposure periods of 0, 7, and 17 days. The titanium levels of the two nanomaterials were the highest in biofilms during the exposure periods. Bioaccumulation factors indicated that TiO2-NP and TiO2-NT were largely transferred from a prey (e.g., biofilm, water dropwort) to its consumer (e.g., nematodes, mud snail). Considering the potential entries of such TiO2 nanomaterials in organisms, their bioaccumulation throughout the food chain should be regarded with great concern in terms of the overall health of the ecosystem.

Effects of methylmercury on epigenetic markers in three model species: mink, chicken and yellow perch.

We previously reported that methylmercury (MeHg) exposure is associated with DNA hypomethylation in the brain stem of male polar bears. Here, we conveniently use archived tissues obtained from controlled laboratory exposure studies to look for evidence that MeHg can disrupt DNA methylation across taxa. Brain (cerebrum) tissues from MeHg-exposed mink (Neovison vison), chicken (Gallus gallus) and yellow perch (Perca flavescens) were analyzed for total Hg levels and global DNA methylation. Tissues from chicken and mink, but not perch, were also analyzed for DNA methyltransferase (DNMT) activity. In mink we observed significant reductions in global DNA methylation in an environmentally-relevant dietary exposure group (1 ppm MeHg), but not in a higher group (2 ppm MeHg). DNMT activity was significantly reduced in all treatment groups. In chicken or yellow perch, no statistically significant effects of MeHg were observed. Dose-dependent trends were observed in the chicken data but the direction of the change was not consistent between the two endpoints. Our results suggest that MeHg can be epigenetically active in that it has the capacity to affect DNA methylation in mammals. The variability in results across species may suggest inter-taxa differences in epigenetic responses to MeHg, or may be related to differences among the exposure scenarios used as animals were exposed to MeHg through different routes (dietary, egg injection), for different periods of time (19-89 days) and at different life stages (embryonic, juvenile, adult).

Elevated mercury exposure and neurochemical alterations in little brown bats (Myotis lucifugus) from a site with historical mercury contamination.

Despite evidence of persistent methylmercury (MeHg) contamination in the South River (Virginia, USA) ecosystem, there is little information concerning MeHg-associated neurological impacts in resident wildlife. Here we determined mercury (Hg) concentrations in tissues of insectivorous little brown bats (Myotis lucifugus) collected from a reference site and a MeHg-contaminated site in the South River ecosystem. We also explored whether neurochemical biomarkers (monoamine oxidase, MAO; acetylcholinesterase, ChE; muscarinic acetylcholine receptor, mAChR; N-methyl-D-aspartate receptor, NMDAR) previously shown to be altered by MeHg in other wildlife were associated with brain Hg levels in these bats. Concentrations of Hg (total and MeHg) in tissues were significantly higher (10-40 fold difference) in South River bats when compared to reference sites. Mean tissue mercury levels (71.9 ppm dw in liver, 7.14 ppm dw in brain, 132 ppm fw in fur) in the South River bats exceed (sub)-clinical thresholds in mammals. When compared to the South River bats, animals from the reference site showed a greater ability to demethylate MeHg in brain (33.1% of total Hg was MeHg vs. 65.5%) and liver (8.9% of total Hg was MeHg vs. 50.8%) thus suggesting differences in their ability to detoxify and eliminate Hg. In terms of Hg-associated neurochemical biomarker responses, interesting biphasic responses were observed with an inflection point between 1 and 5 ppm dw in the brain. In the reference bats Hg-associated decreases in MAO (r = -0.61; p < 0.05) and ChE (r = -0.79; p < 0.01) were found in a manner expected but these were not found in the bats from the contaminated site. Owing to high Hg exposures, differences in Hg metabolism, and the importance of the aforementioned neurochemicals in multiple facets of animal health, altered or perhaps even a lack of expected neurochemical responses in Hg-contaminated bats raise questions about the ecological and physiological impacts of Hg on the bat population as well as the broader ecosystem in the South River.

High levels of uranium in groundwater of Ulaanbaatar, Mongolia.

Water samples collected from 129 wells in seven of the nine sub-divisions of Ulaanbaatar were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) using Clean Lab methods. The levels of many trace elements were found to be low with the average concentrations (ranges in brackets) being 0.9 (<0.1-7.9) µg/L for As; 7.7 (0.12-177) µg/L for Mn; 0.2 (<0.05-1.9)µg/L for Co; 16 (<0.1-686) µg/L for Zn; 0.7 (<0.1-1.8) µg/L for Se; <0.1 (<0.02-0.69) µg/L for Cd; and 1.3 (<0.02-32) µg/L for Pb. The levels of uranium were surprisingly elevated (mean, 4.6 µg/L; range <0.01-57 µg/L), with the values for many samples exceeding the World Health Organization's guideline of 15 µg/L for uranium in drinking water. Local rocks and soils appear to be the natural source of the uranium. The levels of uranium in Ulaanbaatar's groundwater are in the range that has been associated with nephrotoxicity, high blood pressure, bone dysfunction and likely reproductive impairment in human populations. We consider the risk associated with drinking the groundwater with elevated levels of uranium in Ulaanbaatar to be a matter for some public health concern and conclude that the paucity of data on chronic effects of low level exposure is a risk factor for continuing the injury to many people in this city.

Multiple metals exposure and neurotoxic risk in bald eagles (Haliaeetus leucocephalus) from two Great Lakes states.

In the present study, the authors determined concentrations of several elements (As, Cd, Co, Cu, Cr, Mn, Pb, Sb, Zn) in the brains and livers of 46 bald eagles (Haliaeetus leucocephalus) from two Great Lakes states, Michigan and Minnesota. To explore whether exposures are of neurological concern, the authors assessed their associations with neurochemical receptors (N-methyl-D-aspartate [NMDA] and γ-aminobutyric acid A [GABA(A)]) and enzymes (glutamine synthetase [GS] and glutamic acid decarboxylase [GAD]) that play critical roles in vertebrate neurobehavior and reproduction. For most elements, levels in the livers and brains did not differ between region and gender. Hepatic Pb levels averaged 33.1 ppm (dry wt), 30.4% of all carcasses exceeded proposed avian Pb thresholds (>26.4 ppm), and in 30.8% of the birds examined evidence of Pb pellets or fragments was found. Significant changes in the activities of GS and GAD were related to brain concentrations of several metals (Pb, Cd, Co, Cu, Zn). No relationships were found among any of the nine elements and NMDA or GABA(A) receptor levels. When combined with the authors' previous study on these same eagles that showed Hg-associated alterations in GS, GAD, and NMDA receptor levels, the present research suggests that bald eagles are exposed to various elements, especially Pb and Hg, that are capable of causing changes in GABAergic and glutamatergic neurotransmission. The functional significance of these neurochemical changes warrants attention.

Mercury exposure and neurochemical impacts in bald eagles across several Great Lakes states.

In this study, we assessed mercury (Hg) exposure in several tissues (brain, liver, and breast and primary feathers) in bald eagles (Haliaeetus leucocephalus) collected from across five Great Lakes states (Iowa, Michigan, Minnesota, Ohio, and Wisconsin) between 2002-2010, and assessed relationships between brain Hg and neurochemical receptors (NMDA and GABA(A)) and enzymes (glutamine synthetase (GS) and glutamic acid decarboxylase (GAD)). Brain total Hg (THg) levels (dry weight basis) averaged 2.80 µg/g (range: 0.2-34.01), and levels were highest in Michigan birds. THg levels in liver (r(p) = 0.805) and breast feathers (r(p) = 0.611) significantly correlated with those in brain. Brain Hg was not associated with binding to the GABA(A) receptor. Brain THg and inorganic Hg (IHg) were significantly positively correlated with GS activity (THg r(p) = 0.190; IHg r(p) = 0.188) and negatively correlated with NMDA receptor levels (THg r(p) = -0245; IHg r(p) = -0.282), and IHg was negatively correlated with GAD activity (r(s) = -0.196). We also report upon Hg demethylation and relationships between Hg and Se in brain and liver. These results suggest that bald eagles in the Great Lakes region are exposed to Hg at levels capable of causing subclinical neurological damage, and that when tissue burdens are related to proposed avian thresholds approximately 14-27% of eagles studied here may be at risk.

Multiple metals exposure in a small-scale artisanal gold mining community.

Urinary metals were characterized in 57 male residents of a small-scale gold mining community in Ghana. Chromium and arsenic exceeded health guideline values for 52% and 34%, respectively, of all participants. About 10-40% of the participants had urinary levels of aluminum, copper, manganese, nickel, selenium, and zinc that fell outside the U.S. reference range. Exposures appear ubiquitous across the community as none of the elements were associated with occupation, age, and diet.

Rapid methods to detect organic mercury and total selenium in biological samples.

BACKGROUND: Organic mercury (Hg) is a global pollutant of concern and selenium is believed to afford protection against mercury risk though few approaches exist to rapidly assess both chemicals in biological samples. Here, micro-scale and rapid methods to detect organic mercury (< 1.5 ml total sample volume, < 1.5 hour) and total selenium (Se; < 3.0 ml total volume, < 3 hour) from a range of biological samples (10-50 mg) are described. RESULTS: For organic Hg, samples are digested using Tris-HCl buffer (with sequential additions of protease, NaOH, cysteine, CuSO4, acidic NaBr) followed by extraction with toluene and Na2S2O3. The final product is analyzed via commercially available direct/total mercury analyzers. For Se, a fluorometric assay has been developed for microplate readers that involves digestion (HNO3-HClO4 and HCl), conjugation (2,3-diaminonaphthalene), and cyclohexane extraction. Recovery of organic Hg (86-107%) and Se (85-121%) were determined through use of Standard Reference Materials and lemon shark kidney tissues. CONCLUSIONS: The approaches outlined provide an easy, rapid, reproducible, and cost-effective platform for monitoring organic Hg and total Se in biological samples. Owing to the importance of organic Hg and Se in the pathophysiology of Hg, integration of such methods into established research monitoring efforts (that largely focus on screening total Hg only) will help increase understanding of Hg's true risks.

Mortality factors and lead contamination of wild birds from Korea.

Wild birds have frequently been found dead in their natural habitats, but little is known about what ecological stressors may impact health of wild populations. Here, we report the potentially harmful lead (Pb) levels in tissues along with necropsies on 69 individuals of cranes, raptors, and waterfowl found dead between 2000 and 2003 in Korea. In all samples diagnosed, trauma (n = 22), severe emaciation (n = 15), and infectious diseases (n = 11) were identified. In the survey, injury with Pb shot or bullet fragments was associated with three of the deaths; one of three showed lesions suggestive of Pb poisoning in the tissues. Of 69 birds, 12 had >25 ppm dry wt. (equivalent to 8 ppm wet wt.) in liver or kidney, which is known to be a potentially lethal level of Pb in wild birds. Three individuals had hepatic Pb levels of 101.3 ppm (Whooper swan), 120.4 ppm (Great white-fronted goose), and 1,059 ppm (Mandarin duck), with evidence of Pb pellets in their gizzard. This study suggests that many birds examined may be suffering from excessive Pb exposure that may be of health concern with respect to a potential cause of their mortality. The need for additional research is heightened when considering that some migrants are classified as a globally protected species by the International Union for the Conservation of Nature.

Mercury and selenium levels in lemon sharks (Negaprion brevirostris) in relation to a harmful red tide event.

Tissue levels of mercury (Hg; total, organic) and selenium (Se) were assessed in juvenile lemon sharks (Negaprion brevirostris) from Florida nearshore waters collected during a harmful algal bloom (HAB, brevetoxin) event and compared with sharks not exposed to HABs. In all sharks studied, total Hg levels in the muscle were generally present in a molar excess over Se (which may protect against Hg toxicity) and mean muscle Hg levels (0.34 microg/g) exceed safe human consumption guidelines. While there was generally no difference in tissue Hg and Se levels following exposure of sharks to HABs, hepatic Hg levels were significantly lower (56% reduction) in the HAB-exposed sharks compared to controls. As Hg and HABs are globally increasing in scope and magnitude, further work is warranted to assess their interactions and biotic impacts within aquatic ecosystems, especially for a species such as the lemon shark that is classified as a near-threatened species by the International Union for the Conservation of Nature.

A combined ecological and epidemiologic investigation of metal exposures amongst Indigenous peoples near the Marlin Mine in Western Guatemala.

In August 2009 a combined epidemiological and ecological pilot study was conducted to investigate allegations of human rights abuses in the form of exposures to toxic metals experienced by mine workers and Indigenous Mam Mayan near the Marlin Mine in Guatemala. In the human study there were no differences in blood and urine metals when comparing five mine workers with eighteen non-mine workers, and there were no discernible relationships between metal exposures and self-reported health measures in any study group. On the other hand, individuals residing closest to the mine had significantly higher levels of certain metals (urinary mercury, copper, arsenic, and zinc) when compared to those living further away. The levels of blood aluminum, manganese, and cobalt were elevated in comparison to established normal ranges in many individuals; however, there was no apparent relationship to proximity to the mine or occupation, and thus are of unclear significance. In the ecological study, several metals (aluminum, manganese, and cobalt) were found significantly elevated in the river water and sediment sites directly below the mine when compared to sites elsewhere. When the human and ecological results are combined, they suggest that exposures to certain metals may be elevated in sites near the mine but it is not clear if the current magnitude of these elevations poses a significant threat to health. The authors conclude that more robust studies are needed while parallel efforts to minimize the ecological and human impacts of mining proceed. This is critical particularly as the impact of the exposures found could be greatly magnified by expected increases in mining activity over time, synergistic toxicity between metals, and susceptibility for the young and those with pre-existing disease.

Mercury contamination in spotted seatrout, Cynoscion nebulosus: an assessment of liver, kidney, blood, and nervous system health.

Marine fishes in South Florida (Florida Keys-Florida Bay-Everglades region) accumulate higher concentrations of mercury (Hg) in their tissues than similar fishes from other areas of the southeastern U.S., though it is not known whether these elevated levels affect fish health. In this study, we used quantifiable pathological and biochemical indicators to explore Hg-associated differences in marine fish from South Florida, where Hg contamination is high, and from Indian River Lagoon, Florida, which served as a reference area. Hg concentrations in all tissues of mature spotted seatrout (Cynoscion nebulosus) from South Florida were significantly higher than those from Indian River Lagoon and were within the threshold range of those in studies where effects of Hg exposure have been observed. The distribution of Hg among tissues followed the same trend in both areas, with the greatest concentration in kidney tissue, followed by liver, muscle, brain, gonad, and red blood cells. Blood-plasma biochemistry showed that concentrations of iron, inorganic phosphate, lactate dehydrogenase, and aspartate aminotransferase were significantly less in South Florida. Also, fructosamine and alkaline phosphatase were significantly less in South Florida. Liver histology revealed that pyknosis/necrosis, interstitial inflammation, and bile duct hyperplasia were found only in seatrout from South Florida, and steatosis/glycogen was more frequently found in Indian River Lagoon specimens. In renal tissue, interstitial inflammation, glomerular dilatation and thickening, and tubular degeneration and necrosis were more frequently found in South Florida specimens. Changes in the liver cytoskeleton and morphology may explain some of the differences in blood parameters between study areas. Neurochemical analyses showed that brain N-methyl-d-aspartic acid (NMDA) receptors (but not those of muscarinic cholinergic receptors, monoamine oxidase, or acetylcholinesterase) were significantly less in fish from South Florida than from Indian River Lagoon. These findings provide compelling evidence that elevated Hg could cause quantifiable pathological and biochemical changes that might influence the health of spotted seatrout and could also affect other marine fish species.

Neurochemical alterations in lemon shark (Negaprion brevirostris) brains in association with brevetoxin exposure.

Brevetoxins are persistent, bioaccumulative, lipophilic polyether neurotoxins synthesized by Karenia brevis, a harmful algal bloom (HAB) dinoflagellate. Although some marine organisms accumulate potentially harmful levels of brevetoxins, little is known about neurotoxic effects in wild populations. Here, tissue (i.e., liver, kidney, muscle, intestine, gill, brain) brevetoxin levels (as ng PbTx-3 eq/g) and four neurochemical biomarkers (monoamine oxidase, MAO; cholinesterase, ChE; muscarinic cholinergic receptor, mAChR; N-methyl-d-aspartic acid receptor, NMDAR) were compared between eleven lemon sharks collected during a K. brevis bloom and eighteen lemon sharks not exposed to a bloom (controls) in a case-control manner. Brevetoxin levels in tissues were significantly higher in HAB-exposed sharks when compared to controls, and tissue levels (e.g., 277-3112 ng/g in livers, 429-2833 ng/g in gills) in HAB-exposed sharks were comparable to levels detected in a shark (e.g., 1223 ng/g in liver, 930 ng/g in gill) that died presumably of toxin exposure. Further, there were significant correlations between brain brevetoxin levels and ChE activity (r=-0.41; p<0.05), MAO activity (r=-0.37; p<0.05), mAChR levels (r=0.55; p<0.01), and NMDAR levels (r=-0.49; p<0.01). There were no relationships between neurochemical biomarkers and metals (total mercury, methylmercury, selenium). Overall, these results in tissues from free-ranging lemon sharks indicate that ecologically relevant exposures to brevetoxins may cause significant changes in brain neurochemistry. As disruptions to neurochemistry precede structural and functional damage to the nervous system, these results suggest that relevant exposures to HABs may be causing sub-clinical effects in lemon sharks and raise further questions about the ecological and physiological impacts of HABs on marine biota.

Mercury-associated DNA hypomethylation in polar bear brains via the LUminometric Methylation Assay: a sensitive method to study epigenetics in wildlife.

In this paper we describe a novel approach that may shed light on the genomic DNA methylation of organisms with non-resolved genomes. The LUminometric Methylation Assay (LUMA) is permissive for genomic DNA methylation studies of any genome as it relies on the use of methyl-sensitive and -insensitive restriction enzymes followed by polymerase extension via Pyrosequencing technology. Here, LUMA was used to characterize genomic DNA methylation in the lower brain stem region from 47 polar bears subsistence hunted in central East Greenland between 1999 and 2001. In these samples, average genomic DNA methylation was 57.9% +/- 6.69 (SD; range was 42.0 to 72.4%). When genomic DNA methylation was related to brain mercury (Hg) exposure levels, an inverse association was seen between these two variables for the entire study population (P for trend = 0.17). After dichotomizing animals by gender and controlling for age, a negative trend was seen amongst male animals (P for trend = 0.07) but no associations were found in female bears. Such sexually dimorphic responses have been found in other toxicological studies. Our results show that genomic DNA methylation can be quantitatively studied in a highly reproducible manner in tissue samples from a wild organism with a non-resolved genome. As such, LUMA holds great promise as a novel method to explore consequential questions across the ecological sciences that may require an epigenetic understanding.