Dynamics of Trace Element Composition of Bratsk Reservoir Water in Different Periods of Anthropogenic Impact (Baikal Region, Russia).

Bratsk water reservoir is a part of the unified Baikal-Angara water system, the world's largest reservoir of surface drinking water. This study intends to assess the water quality of Bratsk reservoir by examining the spatial and temporal variability of trace element concentrations over periods of changing by the anthropogenic impact. The water samples were collected along the length of the reservoir in 2003, 2008, 2012, and 2018 and analyzed for 12 trace elements (Al, Cr, Mn, Fe, Cu, Zn, As, Cd, Cs, Pb, U, and Hg). The results show that in the zone of maximum anthropogenic impact the concentrations of these elements increased as follows: Al 781.0 µg L-1; Cr 1.43 µg L-1; Fe 1052.4 µg L-1; Cu 3.34 µg L-1; Zn 15.52 µg L-1; Cd 0.691 µg L-1; Cs 0.070 µg L-1; U 1.74 µg L-1; Hg 0.0505 µg L-1. The trace element concentrations in Bratsk reservoir water did not exceed levels set by the international drinking water quality standards. At the same time, the uniqueness of the reservoir requires the use of stricter criteria for water quality control. The trace element concentrations in the water from the source of the Angara river (the only run-off from Lake Baikal) used as reference levels indicate higher levels of metal contamination and pollution indices in Bratsk reservoir water. Over different periods of observation, the contamination factor (CF) showed very high contamination for Al, Mn, Cd, and Cs. PLI indices for 2003 and 2018 classify the majority of water samples as polluted. The NPI values were indicative of severe water contamination by Mn and Pb (all years), Cu (2009), Fe and Cd (2003), Al, Cr, Zn, Cs, and Hg (2003, 2009, 2018). The obtained results showed the importance of using water quality criteria with a view to strengthening the requirements of environmental protection measures in the Baikal region.

Long-Term and Ontogenetic Patterns of Heavy Metal Contamination in Lake Baikal Seals (Pusa sibirica).

Little is known about the history of heavy metal pollution of Russia's Lake Baikal, one of the world's largest lakes and a home to numerous endemic species, including the Baikal Seal, Pusa sibirica. We investigated the history of heavy metal (V, Cu, Zn, Cd, Hg, Tl, Pb, U) pollution in Lake Baikal seals over the past 8 decades. C and N stable isotope analysis (SIA) and laser-ablation ICP-MS of seal teeth were used to examine changes in feeding ecology, heavy metal levels associated with life history events and long-term variation in metal exposure. SIA did not suggest large changes in the feeding ecology of Baikal seals over the past 80 years. LA-ICP-MS analyses revealed element-specific ontogenetic variability in metal concentrations, likely related to maternal transfer, changes in food sources and starvation. Hg and Cd levels in seals varied significantly across the time series, with concentrations peaking in the 1960s - 1970s but then declining to contemporary levels similar to those observed in the 1930s and 1940s. Trends in atmospheric emissions of Hg suggest that local sources as well as emissions from eastern Russia and Europe may be important contributors of Hg to Lake Baikal and that, despite the size of Lake Baikal, its food web appears to respond rapidly to changing inputs of contaminants.

Occurrence and tissue distribution of 33 legacy and novel per- and polyfluoroalkyl substances (PFASs) in Baikal seals (Phoca sibirica).

Per- and polyfluoroalkyl substances (PFASs) are bioaccumulative and associated with adverse effects in both wildlife and humans. The occurrence of 33 PFASs was assessed in the plasma, liver, blubber, and brain of 18 Baikal seals (Phoca sibirica) (16 pups and 2 adult females) from Lake Baikal, Russia (in 2011). Of the 33 congeners analysed for: perfluorooctanosulfonic acid (PFOS), 7 long chain perfluoroalkyl carboxylic acids (C8-C14 PFCAs) and 1 branched PFCA (perfluoro-3,7-dimethyloctanoic acid; P37DMOA) were most frequently detected. The PFASs in plasma and liver with the highest median concentrations were legacy congeners: perfluoroundecanoic acid (PFUnA; plasma: 11.2 ng/g w.w.; liver: 7.36 ng/g w.w.), PFOS (plasma: 8.67 ng/g w.w.; liver: 9.86 ng/g w.w.), perfluorodecanoic acid (PFDA; plasma: 5.13 ng/g w.w.; liver: 6.69 ng/g w.w.), perfluorononanoic acid (PFNA; plasma: 4.65 ng/g w.w.; liver: 5.83 ng/g w.w.) and perfluorotridecanoic acid (PFTriDA; plasma: 4.29 ng/g w.w.; liver: 2.55 ng/g w.w.). PFASs were detected in the brain of Baikal seals, indicating that PFASs cross through the blood-brain barrier. In blubber, the majority of PFASs were detected in low abundance and concentrations. In contrast to legacy PFASs, novel congeners (e.g., Gen X) were either detected infrequently or not found in Baikal seals. The worldwide occurrence of PFASs in pinnipeds was compared and lower median concentrations of PFOS were observed in Baikal seals relative to other pinnipeds. Conversely, similar concentrations of long chain PFCAs were found in Baikal seals compared to other pinnipeds. Furthermore, human exposure was assessed by estimating weekly intakes (EWI) of PFASs through Baikal seal consumption. Although PFASs concentrations were comparatively low relative to other pinnipeds, consumption of Baikal seal could exceed current regulatory guidelines.

Higher mass-independent isotope fractionation of methylmercury in the pelagic food web of Lake Baikal (Russia).

Mercury undergoes several transformations that influence its stable isotope composition during a number of environmental and biological processes. Measurements of Hg isotopic mass-dependent (MDF) and mass-independent fractionation (MIF) in food webs may therefore help to identify major sources and processes leading to significant bioaccumulation of methylmercury (MeHg). In this work, δ(13)C, δ(15)N, concentration of Hg species (MeHg, inorganic Hg), and stable isotopic composition of Hg were determined at different trophic levels of the remote and pristine Lake Baikal ecosystem. Muscle of seals and different fish as well as amphipods, zooplankton, and phytoplankton were specifically investigated. MDF during trophic transfer of MeHg leading to enrichment of heavier isotopes in the predators was clearly established by δ(202)Hg measurements in the pelagic prey-predator system (carnivorous sculpins and top-predator seals). Despite the low concentrations of Hg in the ecosystem, the pelagic food web reveals very high MIF Δ(199)Hg (3.15-6.65‰) in comparison to coastal fish (0.26-1.65‰) and most previous studies in aquatic organisms. Trophic transfer does not influence MIF signature since similar Δ(199)Hg was observed in sculpins (4.59 ± 0.55‰) and seal muscles (4.62 ± 0.60‰). The MIF is suggested to be mainly controlled by specific physical and biogeochemical characteristics of the water column. The higher level of MIF in pelagic fish of Lake Baikal is mainly due to the bioaccumulation of residual MeHg that is efficiently turned over and photodemethylated in deep oligotrophic and stationary (i.e., long residence time) freshwater columns.

Rapid determination of thyroid hormones in blood plasma from Glaucous gulls and Baikal seals by HybridSPE®-LC-MS/MS.

A rapid hybrid solid phase extraction (HybridSPE®) protocol tailored to liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) analysis, was developed for the determination of four thyroid hormones, L-Thyroxine (T4), 3,3',5-triiodo-L-thyronine (T3), 3,3',5'-triiodo-L-thyronine (rT3) and 3,3'-diiodo-L-thyronine (T2) in blood plasma from Glaucous gulls (Larus hyperboreus) and Baikal seals (Phoca sibirica). The use of target analyte specific 13C internal standards allowed quantification to be performed through the standard solvent calibration curves and alleviated the need to perform quantification with matrix match curves. The relative recoveries were 100.0-110.1 % for T4, 99.1-102.2 % for T3, 100.5-108.0 % for rT3, and 100.5-104.6 % for T2. The matrix effects ranged from -1.52 to -6.10 %, demonstrating minor signal suppression during analysis. The method intra-day precision (method repeatability, RSD %, N = 5, k = 1 day) and inter-day precision (method reproducibility, RSD %, N = 10, k = 2 days) at the 1 ng/mL concentration of fortification were 8.54-15.4 % and 15.4-24.8 %, respectively, indicating acceptable chromatographic peak stabilities for all target THs even at trace level concentrations. The method limit of detection (LOD) for T4, T3, rT3 and T2 was 0.17, 0.16, 0.30 and 0.17 ng/mL, respectively. The HybridSPE® protocol was simple and rapid (<1 min) upon application, while the HybridSPE® cartridge did not require (as in classical SPE cartridges) any additional equilibration nor conditioning step prior sample loading. A total of 46 blood plasma samples, 30 samples collected from Glaucous gulls and 16 samples collected from Baikal seals, were analyzed for thyroid hormones to demonstrate the applicability of the developed method in these wildlife species. The concentrations of T4 and T3 in blood plasma from the Glaucous gulls were 5.95-44.2 and 0.37-5.61 ng/mL, respectively, whereas those from Baikal seals were 3.57-46.5 and 0.45-2.07 ng/mL, respectively. In both species, rT3 demonstrated low detection rate, while T2 was not detected. Furthermore, cross-array comparison between the HybridSPE®-LC-MS/MS protocol and an established routine radioimmunoassay (RIA) kit-based method was performed for T4 and T3 concentrations from selected Baikal seal plasma samples.

Tracing sources and bioaccumulation of mercury in fish of Lake Baikal--Angara River using Hg isotopic composition.

This study presents the determination and comparison of isotopic compositions of Hg in sediments, plankton, roach, and perch of two freshwater systems in the Lake Baikal-Angara River aquatic ecosystem: the man-made Bratsk Water Reservoir contaminated by Hg from a chlor-alkali factory and the noncontaminated Lake Baikal. Isotopic ratios of biota exhibit both significant mass-independent fractionation (MIF) (Δ(199)Hg from 0.20 to 1.87‰) and mass-dependent fractionation (MDF) (δ(202)Hg from -0.97 to -0.16‰), whereas sediments exhibit high MDF (δ(202)Hg from -1.99 to -0.83‰) but no MIF. δ(15)N and δ(13)C are correlated with methylmercury in organisms from both sites, indicating bioaccumulation and biomagnification through food webs of both regions. Combining this with isotopic composition of samples shows that δ(202)Hg increases with the trophic level of organisms and also with methylmercury in fish from Lake Baikal. This study demonstrates that MIF in fish samples from Bratsk Water Reservoir allow to trace anthropogenic Hg, since fish with the highest levels of Hg in muscle have the same isotopic composition as the sediment in which anthropogenic Hg was deposited. Less contaminated fish do not exhibit this anthropogenic signature accumulating relatively lower Hg amount from the contaminated sediments. This work reveals that Hg isotopic composition can be used to track the contribution of anthropogenic sources in fish from a contaminated lake.

The Phenomenon of Compensatory Cell Proliferation in Olfactory Epithelium in Fish Caused by Prolonged Exposure to Natural Odorants.

It was previously shown that activation of the processes of neurogenesis in the olfactory epithelium (OE) can be caused after intranasal administration of toxic or neurotrophic factors, after axon transection, or as a result of bulbectomy. Our study showed for the first time that a significant increase in olfactory cell renewal can also occur in animals due to periodic chemostimulation with natural odorants (amino acids and peptides) for 15 days. Using electron and laser confocal microscopy in fish (Paracottus knerii (Cottidae), Dybowski, 1874) from Lake Baikal, we showed that periodic stimulation of aquatic organisms with a water-soluble mixture of amino acids and peptides causes stress in OE, which leads to programmed death cells and compensatory intensification of their renewal. We estimated the level of reactive oxygen species, number of functionally active mitochondria, intensity of apoptosis processes, and mitosis activity of cells in the OE of fish in the control group and after periodic natural odorants exposure. This study showed that new stem cells are activated during enhanced odor stimulation and subsequent degenerative changes in the cells of the sensory apparatus. Those new activated stem cells are located in previously proliferatively inactive regions of OE that become involved in compensatory processes for the formation of new cells.

Rearrangement of Actin Microfilaments in the Development of Olfactory Receptor Cells in Fish.

At present, it remains poorly understood how the olfactory neuron migrates through the thick neuroepithelium during its maturation from a stem cell and how it develops a specific sensitivity to environmental odorants after maturation. We investigated the cytochemical features associated with the development of olfactory cells before and after the incorporation of dendrites into the surface of the olfactory epithelium. Using cytochemical staining for the actin cytoskeleton and other cell components, we found that immature neurons acquire a streamlined shape that resembles a «hot-dog¼ during their migration: a dense layer of actin microfilaments forms beneath the surface membrane of the growing dendrite, and the bulk of the nuclear material moves inside this layer. We have found that when the cell makes contact with its environment, the dendritic terminal develops a wide actin layer, inside which a pore is formed. It is assumed that the functional receptors of odorants generate across this pore the first intracellular signal from environmental water-soluble odorants. These data illustrate the important role of the cytoskeleton in the differentiation of olfactory cells.

Past and present mercury accumulation in the Lake Baikal seal: Temporal trends, effects of life history, and toxicological implications.

Despite global efforts to reduce anthropogenic mercury (Hg) emissions, the timescale and degree to which Hg concentrations in the environment and biota respond to decreased emissions remain challenging to assess or predict. In the present study we characterize long-term trends and life-history patterns in Hg accumulation and toxicological implications of Hg contamination for a freshwater seal from one of the world's largest lakes (Lake Baikal, Siberia, Russia) using contemporary tissues and archival teeth. Stable isotope analysis and Hg analyses of soft tissues (muscle, liver, kidney, blood, brain, heart) and teeth from 22 contemporary seals revealed rapid changes in diet and Hg accumulation in the first year of life with a stable diet and increase in tissue Hg throughout the rest of life. Although maternal transfer of Hg was an important source of Hg to seal pups, reproduction and lactation by female seals did not appear to result in sex-related differences in Hg concentrations or age-related accumulation in adult seals. Based on Hg analysis of archival teeth (n = 114) and reconstructed values for soft tissues, we also assessed temporal trends in seal Hg between the years 1960 and 2013. Seal Hg concentrations in hard (teeth) and soft (e.g., muscle, liver) tissues were highest in the 1960s and 1970s, followed by a decrease. The decline in seal Hg concentrations in recent decades was most likely driven by a reduction in Hg inputs to the lake, suggesting that global and regional efforts to reduce Hg emissions have been successful at reducing ecosystem and human health risks posed by Hg in Lake Baikal. Environ Toxicol Chem 2018;37:1476-1486. © 2018 SETAC.

Relationships and bioaccumulation of chemical elements in the Baikal seal (Phoca sibirica).

Concentrations of Al, Ba, Cd, Cu, Fe, Mn, Mo, Si, Sr, Zn, Ca, K, Mg, Na and P in the livers of Baikal seal, plankton, zoobenthos, and fish, constituting the food sources for the seals, were determined by ICP-MS and ICP-AES. The accumulation of elements in the liver of seals, affected by internal and external (environmental) factors, was assessed by multidimensional (ANOVA, FA) and correlation analyses. FA has enabled identification of abiotic and biotic factors responsible for the accumulation of elements in the livers of Baikal seals. Significant influence of sex and development stage of the seals analysed on hepatic concentrations of some elements was found. The observed differences in element concentrations between pups, males and females could be attributable to the reproductive cycle of this species. ANOVA showed differences in concentrations of Fe, Zn, Cu and Cd in seals from the three separate basins of the lake. BMFs suggest biomagnification of Fe and Zn in the fish-seal trophic link.

Natural Hg isotopic composition of different Hg compounds in mammal tissues as a proxy for in vivo breakdown of toxic methylmercury.

In the last decade, specific attention has been paid to total mercury (HgT) stable isotopic composition, especially in natural samples such as aquatic organisms, due to its potential to track the cycle of this toxic element in the environment. Here, we investigated Hg Compound Specific stable Isotopic Composition (CSIC) of natural inorganic Hg (iHg) and methylmercury (MMHg) in various tissues of aquatic mammals (Beluga whale from the Arctic marine environment and seals from the freshwater lake Baikal, Russia). In seals' organs the variation in mass dependent fractionation (MDF, δ(202)Hg) for total Hg was significantly correlated to the respective fraction of iHg and MMHg compounds, with MMHg being enriched by ∼ 3‰ in heavier isotopes relative to iHg. On the other hand, we observe insignificant variation in Hg mass independent isotope fractionation (MIF, Δ(199)Hg) among iHg and MMHg in all organs for the same mammal species and MMHg in prey items. MIF signatures suggest that both MMHg and iHg in aquatic mammals have the same origin (i.e., MMHg from food), and are representative of Hg photochemistry in the water column of the mammal ecosystem. MDF signatures of Hg compounds indicate that MMHg is demethylated in vivo before being stored in the muscle, and the iHg formed is stored in the liver, and to a lesser extent in the kidney, before excretion. Thus, Hg CSIC analysis in mammals can be a powerful tool for tracing the metabolic response to Hg exposure.

Differential bioaccumulation of potentially toxic elements in benthic and pelagic food chains in Lake Baikal.

Lake Baikal is located in eastern Siberia in the center of a vast mountain region. Even though the lake is regarded as a unique and pristine ecosystem, there are existing sources of anthropogenic pollution to the lake. In this study, the concentrations of the potentially toxic trace elements As, Cd, Pb, Hg, and Se were analyzed in water, plankton, invertebrates, and fish from riverine and pelagic influenced sites in Lake Baikal. Concentrations of Cd, Hg, Pb and Se in Lake Baikal water and biota were low, while concentrations of As were similar or slightly higher compared to in other freshwater ecosystems. The bioaccumulation potential of the trace elements in both the pelagic and the benthic ecosystems differed between the Selenga Shallows (riverine influence) and the Listvenichnyi Bay (pelagic influence). Despite the one order of magnitude higher water concentrations of Pb in the Selenga Shallows, Pb concentrations were significantly higher in both pelagic and benthic fish from the Listvenichnyi Bay. A similar trend was observed for Cd, Hg, and Se. The identified enhanced bioavailability of contaminants in the pelagic influenced Listvenichnyi Bay may be attributed to a lower abundance of natural ligands for contaminant complexation. Hg was found to biomagnify in both benthic and pelagic Baikal food chains, while As, Cd, and Pb were biodiluted. At both locations, Hg concentrations were around seven times higher in benthic than in pelagic fish, while pelagic fish had two times higher As concentrations compared to benthic fish. The calculated Se/Hg molar ratios revealed that, even though Lake Baikal is located in a Se-deficient region, Se is still present in excess over Hg and therefore the probability of Hg induced toxicity in the endemic fish species of Lake Baikal is assumed to be low.

Lake Baikal Endemic Sculpins (Cottoidei): A Promising Model to Study Adaptive Plasticity of Blood Cholesterol Metabolism

We analyzed the blood lipid spectra in four closely related sculpin (Cottoidei) species endemic to Lake Baikal. These data characterize the Baikal sculpins as a set of model organisms for studying the adaptive plasticity of cholesterol metabolism and also mechanisms of resistance to the development of dyslipidemia and atherosclerosis.