Mercury in European Blushers,Amanita rubescens,mushrooms and topsoils: Bioconcentration potential and intake assessment

Total mercury content has been determined in fruiting bodies of European Blushers and topsoils collected from 11 sites across Poland in 2006-2008. Mercury analysis was carried out using a validated analytical method and cold-vapour atomic absorption (CV-AAS). The European Blusher effectively accumulated mercury in fruiting bodies. The mean values of total mercury in caps of European Blushers from background (uncontaminated) areas were from 0.22 to 1.0 (0.067-3.2) and in stipes from 0.16 to 0.65 (0.071-2.7) μg/g dry weight. In topsoil beneath to fruiting bodies, the median Hg concentration at 10 sites in Northern Poland varied between 0.030 and 0.072 (0.0096–0.19) μg/g dw, and in one site in Southern Poland was 0.20 (0.079–0.34) μg/g dw. Data on Hg in European Blushers from different countries were reviewed. The mean concentrations of total Hg in caps of European Blushers from two “pristine” sites in northern part of Poland were ～1.0 μg/g dw. A meal made with 300–500 g of fresh caps of European Blushers collected at such sites (assuming 90% water content in caps) can result in Hg intake of 0.0003–0.0005 mg Hg/kg bm (assuming a 60 kg bm), which is a dose equipotent to a new provisional tolerable weekly intake (PTWI) value set for inorganic Hg.     

Mercury in certain boletus mushrooms from Poland and Belarus

This paper reports the results of the study of Hg contents of four species of Boletus mushroom (Boletus reticulatus Schaeff. 1763, B. pinophilus Pilát & Dermek 1973, B. impolitus Fr. 1838 and B. luridus Schaeff. 1774) and the surface soils (0–10 cm layer, ～100 g) samples beneath the mushrooms from ten forested areas in Poland and Belarus by cold-vapour atomic absorption spectroscopy. The ability of the species to bioconcentrate Hg was calculated (as the BCF) while Hg intakes from consumption of these mushroom species were also estimated. The median Hg content of the caps of the species varied between 0.38 and 4.7 mg kg^?1 dm; in stipes between 0.13 and 2.5 mg kg^?1 dm and in the mean Hg contents of soils varied from 0.020 ± 0.01 mg kg^?1 dm to 0.17 ± 0.10 mg kg^?1 dm which is considered as “background” Hg level. The median Hg content of caps of B. reticulatus and B. pinophilus were up to 4.7 and 3.6 mg kg^?1 dm, respectively, and they very efficiently bioaccumulate Hg with median BCF values of up to 130 for caps and 58 for stipes. The caps and stipes of these mushrooms if eaten will expose consumer to elevated dose of total Hg estimated at 1.4 mg for caps of Boletus reticulatus from the Kacze ??gi site, which is a nature reserve area. Nevertheless, the occasional consumption of the valued B. reticulatus and B. pinophilus mushrooms maybe safe.     

Bioconcentration and depuration of endosulfan sulfate in mosquito fish (Gambusia affinis)

Endosulfan is an insecticide which has been widely used in agriculture. The technical grade material consists of two isomers (alpha and beta). Under natural environmental conditions, endosulfan is metabolized through oxidation and the main metabolite in the environment is endosulfan sulfate. Most ecotoxicology research has been conducted with technical grade endosulfan to determine effects on non-target aquatic organisms. Little data on the effects of endosulfan sulfate on aquatic organisms are available in the literature. This study characterizes endosulfan sulfate bioconcentration and depuration in mosquito fish (Gambusia affinis). During the study, G. affinis was exposed to an environmentally relevant endosulfan sulfate concentration of 0.25 μg L⁻¹ for 5 weeks (uptake phase) followed by a 3-week period (depuration phase) in clean water. This study found that G. affinis bioconcentrated endosulfan sulfate. During the exposure phase, fish tissue concentrations of endosulfan sulfate increased with time up to 730 μg kg⁻¹ dw or 215 μg kg⁻¹ ww. The bioconcentration data followed Michaelis-Menten kinetics better than the one-compartment first order kinetics (1-CFOK). Using these models, the bioconcentration factors for endosulfan sulfate-exposed G. affinis were from 687 to 888  L kg⁻¹ in wet weight or 2263 to 2936 L kg⁻¹ in dry weight. During the depuration phase, endosulfan sulfate concentrations in tissue significantly decreased and the data followed first order kinetics. The half-life of endosulfan sulfate in G. affinis was about 9 d. There was no significant difference in standard length or weight between control and exposed fish. The growth data followed the von Bertalanffy growth model. However, the condition factor of exposed fish increased with time during the exposure phase.     

Inorganic elemental concentrations in birch bolete mushroom (Leccinum scabrum) and top soil: contamination profiles,bioconcentation and annual variations

Abstract

Analysis of inorganic and organic contaminants in foodstuffs aids in understanding the human exposure to these compounds via consumption. In this study, an edible mushroom species (Leccinum scabrum) and top soil samples were analysed for essential and toxic substances including phosphorus and inorganic elements over a period of three fruiting seasons. Analysis of silver (Ag), aluminium (Al), barium (Ba), calcium (Ca), cadmium (Cd), cobalt (Co), copper (Cu), iron (Fe), mercury (Hg), potassium (K), magnesium (Mg), manganese (Mn), sodium (Na), nickel (Ni), phosphorus (P), lead (Pb), rubidium (Rb), strontium (Sr) and zinc (Zn) in mushrooms and topsoil were performed using inductively coupled plasma optical emission spectroscopy (ICP-OES) with ultrasonic cross flow nebulizer. Total mercury was determined by cold-vapour atomic absorption spectroscopy (CV-AAS). The results exhibited wide variation in concentrations of metals between soil and mushroom (cap and stipes) during three fruiting seasons. Positive bioconcentration factors (BFCs) indicate on bioaccumulation of several metals including, Cd, Cu, Hg, K, Mg, Na, P, Rb and Zn in caps and stipes of fruitbodies of this mushroom, while other metals such as Al, Ba, Ca, Co, Fe, Mn, Ni, Pb and Sr were not exhibiting significant positive BFCs. Over a period studied, the caps were characterised by different (p?<?0.05) concentrations of Al, Co, Cu, Hg, Mn, Ni, P, Pb and Sr. Contamination profiles, temporal fluctuations, BCFs should be taken into consideration when assessing the nutritional value of this mushroom.     

Content and bioconcentration factors of mercury by Parasol Mushroom Macrolepiota procera

The carpophores of Parasol Mushroom and underlying soil substrate collected from several unpolluted and spatially distant sites across Poland were examined to know content and bioconcentration potential of mercury by this species. The total mercury content of the caps of Parasol Mushroom for the particular sites ranged from 1.1 ± 1.0 to 8.4 ± 7.4 μ g/g dry matter (total range from 0.05 to 22 μ g/g dm), while in the stalks were from 0.53 ± 0.27 to 6.8 ± 7.1 μ g/g dm (total range from 0.078 to 20 μ g/g dm). A top soil layer (0–10 cm) showed baseline mercury concentration from 0.022 ± 0.011 to 0.36 ± 0.16 μ g/g dm (total range from 0.010 to 0.54 μg/g dm). Parasol Mushroom is an effective mercury accumulator in the carpophores and bioconcentration factor (BCF) values of this element in the caps and depending on the sampling site ranged from 16 ± 6 to 220 ± 110 (total range from 0.52 to 470), while for the stalks were from 7.6 ± 2.6 to 130 ± 96 (total range from 0.52 to 340). It seems reasonable to state that tolerance (maximum allowable concentration) of the total mercury in a single cap of Parasol Mushroom at unpolluted areas should not exceed 25 μ g/g dm. A value greater then 25 μ g/g dm will imply an elevated content due to site pollution problems. Nevertheless, knowledge on highly toxic methylmercury content and its fraction in the total mercury content of Parasol Mushroom is lacking.     

Bioaccumulation Kinetics and Bioconcentration Factor of Chlorinated Pesticides in Tissues of Puntius ticto (Ham.)

Abstract

Bioaccumulation kinetics and bioconcentration factor (BCF) of chlorinated pesticides like Aldrin, Dieldrin, Benzene hexachloride (BHC), and Dichloro-diphenyl-dichloro-ethane (DDT) in fish tissues of Puntius ticto was studied in detail in a continuous fed system. The bioconcentration process is summarized by using a first order uptake model and the steady-state BCF is calculated based on the 30 days exposure. Rate of bioaccumulation of DDT was maximum of 4.6432 µg g^?1 wet weight per day in liver tissue whereas it was minimum of 0.0002 µg g^?1 wet weight per day in case of Dieldrin in the muscle tissue among the pesticides. It was observed that DDT showed maximum BCF of 89,010 in case of liver tissue of the fish exposed to 30 days. The regression coefficient (r ²) between pesticide concentration and exposure time varied between 0.6212 and 0.9817 indicating high correlation. Based on actual calculated BCF values, the octanol–water partition coefficient (K _ow) values were predicted. In order to prove the hydrophobic property of chlorinated compounds and its affinity towards lipid, the K _ow is predicted. Results showed that pesticide burden differ from tissue to tissue and can be correlated to the lipid content, size, exposure time, and species.     

Bioconcentration of triclosan, methyl-triclosan, and triclocarban in the plants and sediments of a constructed wetland

Constructed wetlands are a potential method for the removal of two pharmaceutical and personal care products from wastewater effluent. Triclosan (TCS; 5-chloro-2-[2,4-dichlorophenoxy]phenol) and triclocarban (TCC; 3,4,4′-trichlorocarbanillide) are antimicrobial agents added to a variety of consumer products whose accumulation patterns in constructed wetlands are poorly understood. Here, we report the accumulation of TCS, its metabolite methyl-triclosan (MTCS; 5-chloro-2-[2,4-dichlorophenoxy]), and TCC in wetland plant tissues and sediments. Three wetland macrophytes: Typha latifolia, Pontederia cordata, and Sagittaria graminea were sampled from a constructed wetland in Denton, Texas, USA. MTCS concentrations were below the method detection limit (MDL) for all species. TCS root tissue concentrations in T. latifolia were significantly greater than root concentrations in P. cordata (mean ± SE in ng g⁻¹: 40.3 ± 11.3 vs. 15.0 ± 1.9, respectively), while for TCC, shoot tissue concentrations in S. graminea were significantly greater than in T. latifolia (22.8 ± 9.3 vs. 9.0 (MDL), respectively). For both TCS and TCC, T. latifolia root tissue concentrations were significantly greater than shoot concentrations (TCS: 40.3 ± 11.3 vs. 17.2 ± 0.2, TCC: 26.0 ± 3.6 vs. 9.0, (MDL)). TCC concentrations in P. cordata roots were significantly greater than in shoots (34.4 ± 5.3 vs. 15.4 ± 2.8, respectively). TCS concentrations in T. latifolia roots and sediments and TCC concentrations in sediments generally decreased from wetland inflow to outflow. To our knowledge, this is the first study documenting species and tissue specific differences in the accumulation of TCS and TCC in plants from an operational constructed wetland. The species specific differences in bioaccumulation suggest TCS and TCC removal from constructed wetlands could be enhanced through targeted plantings.     

Bioconcentration of ibuprofen in fathead minnow (Pimephales promelas) and channel catfish (Ictalurus punctatus)

Pharmaceutical products and their metabolites are being widely detected in aquatic environments and there is a growing interest in assessing potential risks of these substances to fish and other non-target species. Ibuprofen is one of the most commonly used analgesic drugs and no peer-reviewed laboratory studies have evaluated the tissue specific bioconcentration of ibuprofen in fish. In the current study, fathead minnow (Pimephales promelas) were exposed to 250 μg L⁻¹ ibuprofen for 28 d followed by a 14 d depuration phase. In a minimized bioconcentration test design, channel catfish (Ictalurus punctatus) were exposed to 250 μg L⁻¹ for a week and allowed to depurate for 7 d. Tissues were collected during uptake and depuration phases of each test and the corresponding proportional and kinetic bioconcentration factors (BCFs) were estimated. The results indicated that the BCF levels were very low (0.08-1.4) implying the lack of bioconcentration potential for ibuprofen in the two species. The highest accumulation of ibuprofen was observed in the catfish plasma as opposed to individual tissues. The minimized test design yielded similar bioconcentration results as those of the standard test and has potential for its use in screening approaches for pharmaceuticals and other classes of chemicals.     

Accumulation of metallic elements by Amanita muscaria from rural lowland and industrial upland regions

ABSTRACT

This study was carried out on the accumulation and occurrence of Ag, Al, Ba, Ca, Cd, Co, Cu, Fe, Hg, K, Mg, Mn, Na, Rb, Sr and Zn in the mushroom Amanita muscaria and forest topsoil from two lowland sites in the Tuchola Pinewoods in the north-central region and an upland site in the ?wietokrzyskie Mountains in the south-central region of Poland. Topsoil from the upland location showed Ag, Al, Ba, Ca, Cd, Co, Cu, Fe, Hg, Na and Zn at significantly higher concentration levels (pseudo-total fraction and often also the labile or extractable fraction) than at both lowland locations, where topsoil was richer in Mg, and similar in Rb. Amanita muscaria from the upland region differed from individuals collected in the lowland sites by higher concentration levels of Cd, Cu, Hg and Mn in caps. This could be related to higher concentration levels of the metallic elements in topsoil in the upland region. On other side, A. muscaria from the upland site was poorer in Co and Fe in caps, and in Ca, Co, Fe and Sr in stipes. In spite of the differences in content of the geogenic metallic elements in topsoil between the lowland and upland locations, A. muscaria from both regions was able to regulate uptake and accumulation of Ag, Al, Ba, Ca, K, Mg, Na, Rb and Zn, which were at similar concentration levels in caps but not necessarily in stipes.     

Molecular diversity of arbuscular mycorrhizal fungi in relation to soil chemical properties and heavy metal contamination

Abundance and diversity of arbuscular mycorrhizal fungi (AMF) associated with dominant plant species were studied along a transect from highly lead (Pb) and zinc (Zn) polluted to non-polluted soil at the Anguran open pit mine in Iran. Using an established primer set for AMF in the internal transcribed spacer (ITS) region of rDNA, nine different AMF sequence types were distinguished after phylogenetic analyses, showing remarkable differences in their distribution patterns along the transect. With decreasing Pb and Zn concentration, the number of AMF sequence types increased, however one sequence type was only found in the highly contaminated area. Multivariate statistical analysis revealed that further factors than HM soil concentration affect the AMF community at contaminated sites. Specifically, the soils’ calcium carbonate equivalent and available P proved to be of importance, which illustrates that field studies on AMF distribution should also consider important environmental factors and their possible interactions.     

Evaluation of vulnerability of Suillus variegatus and Suillus granulatus mushrooms to sequester mercury in fruiting bodies

This work determined the mercury (Hg) contents and bioconcentration potential of two Suillus mushrooms, and the probable dietary intake of this element from a mushroom meal. The determination of total Hg content of fungal and soil samples was performed using cold-vapour atomic absorption spectroscopy by a direct sample thermal decomposition coupled with gold wool trap of Hg and its further desorption and quantitative measurement at a wavelength of 253.7 nm. The median values of Hg contents (mg kg^?1 dry biomass) in 213 specimens of S. variegatus from 12 background areas varied widely from 0.087 to 0.51 for caps and from 0.041 to 0.24 for stipes. In 52 specimens of S. granulatus, the Hg contents ranged from 0.30 to 0.41 for caps and from 0.058 to 0.14 for stipes. Both species could be classified as moderate accumulators of Hg and the median bioconcentration factor values ranged from 7.0 to 14 (caps) and 2.1 to 13 (stipes) for S. variegatus and 9.5 (caps) and 1.3 (stipes) for S. granulatus. The estimated intake rates of Hg with the consumption of 300-g caps were from 0.0026 to 0.015 per capita or from 0.000037 to 0.00022 mg kg^?1 body mass and this do not indicate any cause for concern associated with eating a meal once or more in a week during the mushrooming season.     

Model evaluation of the phytoextraction potential of heavy metal hyperaccumulators and non-hyperaccumulators

Evaluation of the remediation ability of zinc/cadmium in hyper- and non-hyperaccumulator plant species through greenhouse studies is limited. To bridge the gap between greenhouse studies and field applications for phytoextraction, we used published data to examine the partitioning of heavy metals between plants and soil (defined as the bioconcentration factor). We compared the remediation ability of the Zn/Cd hyperaccumulators Thlaspi caerulescens and Arabidopsis halleri and the non-hyperaccumulators Nicotiana tabacum and Brassica juncea using a hierarchical linear model (HLM). A recursive algorithm was then used to evaluate how many harvest cycles were required to clean a contaminated site to meet Taiwan Environmental Protection Agency regulations. Despite the high bioconcentration factor of both hyperaccumulators, metal removal was still limited because of the plants' small biomass. Simulation with N. tabacum and the Cadmium model suggests further study and development of plants with high biomass and improved phytoextraction potential for use in environmental cleanup.     

Correlation between contamination of the rural environment with mercury and occurrence of leukaemia in men and cattle

A statistically significant positive rectilinear correlation was found between the use of fungicides which contain mercury and the occurrence of leukaemia among the inhabitants of rural areas in the Cracow region (Southern Poland). Higher mercury content was detected in cattle food used by leukaemic farmers, when compared with the healthy ones. A positive correlation was also observed between the contamination of environment with seed dressings containing mercury and a frequency of lymphocytic leukaemia in cattle. In the course of leukaemia, there was a tendency towards a slower excretion of mercury and its accumulation in the organism, e.g.in hair.     

混汞法采金地区的汞污染研究进展

混汞法是一种设备简单、操作简便且应用历史悠久的提金方法 ,在世界范围内得到了普遍使用 ,同时也导致了严重的汞污染。本文介绍了混汞法的流程以及汞的释放过程和释放因子 ;综述了应用混汞法提金的不同国家和地区 ,对由此引发的大气、水体、生物 (包括人体 )和土壤汞污染的研究现状 ,并讨论了采金地区的环境管理政策     

Temporal variability in 20 chemical elements content of Parasol Mushroom (Macrolepiota procera) collected from two sites over a few years

Mature specimens of Parasol Mushroom were collected annually in the outskirts of the Siemiany (2000–2003) and Rafa (2001–2003) sites in the northern part of Poland to examine temporal variations and similarities in the composition of 20 chemical elements. Analysis was done under the same condition and using well-validated analytical methods. Elements were determined by inductively coupled plasma–atomic emission spectroscopy and cold vapour–atomic absorption spectroscopy (Hg). The ranges of Ag, Al, Ba, Ca, Cd, Co, Cu, Cr, Fe, Hg, K, Mg, Mn, Na, Ni, P, Pb, Rb, Sr and Zn concentrations in the caps of fruiting bodies were similar (p > 0.05; Mann-Whitney U test) for both geographically distant sites, and these specimens from Rafa were more contaminated with Pb (p < 0.05; Mann-Whitney U test). The annual collections of caps in the Siemiany site varied in Ag, Al, Ba, Ca, Cd, Co, Cu, Fe, Hg, Na, Rb and Sr and contents (0.05 < p < 0.001), while they were similar in Cr, K, Mg, Mn, Ni, P, Pb and Zn (p > 0.05; Mann-Whitney U test). The annual collections of specimens from the Rafa site varied in contents of Ag, Al, Ba, Ca, Fe, Hg, K, Mg, Mn, P, Rb and Zn (p > 0.05), while they were similar in Cd, Co, Cr, Cu, Na, Ni, Pb and Sr (p < 0.05). The results of this study imply that metallic elements content of Parasol Mushroom collected at the same undisrupted sites, and hence keeping the same geochemical condition for mushroom development and fructification (the same stands and probably the same mycelia), can fluctuate over the years or the life-span of mycelium. Hence, when assessing the nutritional value of essential metallic elements and status of non-essential or toxic metallic elements in Parasol's Mushroom caps (and probably also of other mushrooms species) to man, the possible fluctuation in contents over time have to be taken into account.     

Dynamics of mercury fluxes and their controlling factors in large Hg-polluted floodplain areas

Environmental pollution by mercury (Hg) is a considerable environmental problem world-wide. Due to the occurrence of Hg volatilization from their soils, floodplains can function as an important source of volatile Hg. Soil temperature and soil water content related to flood dynamics are considered as important factors affecting seasonal dynamics of total gaseous mercury (TGM) fluxes. We quantified seasonal variations of TGM fluxes and conducted a laboratory microcosm experiment to assess the effect of temperature and moisture on TGM fluxes in heavily polluted floodplain soils. Observed TGM emissions ranged from 10 to 850 ng m⁻² h⁻¹ and extremely exceeded the emissions of non-polluted sites. TGM emissions increased exponentially with raised air and soil temperatures in both field (R²: 0.49-0.70) and laboratory (R²: 0.99) experiments. Wet soil material showed higher TGM fluxes, whereas the role of soil water content was affected by sampling time during the microcosm experiments.     

Biodegradation and detoxification of olive mill wastewater by selected strains of the mushroom genera Ganoderma and Pleurotus

Thirty-nine white-rot fungi belonging to nine species of Agaricomycotina (Basidiomycota) were initially screened for their ability to decrease olive-mill wastewater (OMW) phenolics. Four strains of Ganoderma australe, Ganoderma carnosum, Pleurotus eryngii and Pleurotus ostreatus, were selected and further examined for key-aspects of the OMW biodegradation process. Fungal growth in OMW-containing batch cultures resulted in significant decolorization (by 40-46% and 60-65% for Ganoderma and Pleurotus spp. respectively) and reduction of phenolics (by 64-67% and 74-81% for Ganoderma and Pleurotus spp. respectively). COD decrease was less pronounced (12-29%). Cress-seeds germination increased by 30-40% when OMW was treated by Pleurotus strains. Toxicity expressed as inhibition of Aliivibrio fischeri luminescence was reduced in fungal-treated OMW samples by approximately 5-15 times compared to the control. As regards the pertinent enzyme activities, laccase and Mn-independent peroxidase were detected for Ganoderma spp. during the entire incubation period. In contrast, Pleurotus spp. did not exhibit any enzyme activities at early growth stages; instead, high laccase (five times greater than those of Ganoderma spp.) and Mn peroxidases activities were determined at the end of treatment. OMW decolorization by Ganoderma strains was strongly correlated to the reduction of phenolics, whereas P. eryngii laccase activity was correlated with the effluent’s decolorization.     

Impact of gold mining associated with mercury contamination in soil,biota sediments and tailings in Kenya

This work considered the environmental impact of artisanal mining gold activity in the Migori–Transmara area (Kenya). From artisanal gold mining, mercury is released to the environment, thus contributing to degradation of soil and water bodies. High mercury contents have been quantified in soil (140 μg kg^?1), sediment (430 μg kg^?1) and tailings (8,900 μg kg^?1), as expected. The results reveal that the mechanism for transporting mercury to the terrestrial ecosystem is associated with wet and dry depositions. Lichens and mosses, used as bioindicators of pollution, are related to the proximity to mining areas. The further the distance from mining areas, the lower the mercury levels. This study also provides risk maps to evaluate potential negative repercussions. We conclude that the Migori–Transmara region can be considered a strongly polluted area with high mercury contents. The technology used to extract gold throughout amalgamation processes causes a high degree of mercury pollution around this gold mining area. Thus, alternative gold extraction methods should be considered to reduce mercury levels that can be released to the environment.     

Historical mercury contamination in sediments and catchment soils of Diss Mere, UK

A 5.3 m sediment core and soil samples were taken from Diss Mere and its catchment. The sediment core was dated and Hg analysed on the sediment and soil samples. The Hg record of the sediment core shows that Diss Mere has been contaminated for the past thousand years and the historical trends in sediment contamination are in good agreement with the development of the weaving industry in Diss and hemp cultivation in the region. Mercury contamination in Diss Mere has been significant and reached a peak in the mid-19th century with sediment Hg concentrations over 50 μg g⁻¹. Elevated Hg concentrations were also found in contemporary soils in residential areas with former industrial land use. Although local hemp cultivation and the traditional weaving industry were abandoned a hundred years ago, Hg contamination caused by these activities still exists in the catchment, and affects the lake.     

History of mercury use and environmental contamination at the Oak Ridge Y-12 Plant

Between 1950 and 1963 approximately 11 million kilograms of mercury (Hg) were used at the Oak Ridge Y-12 National Security Complex (Y-12 NSC) for lithium isotope separation processes. About 3% of the Hg was lost to the air, soil and rock under facilities, and East Fork Poplar Creek (EFPC) which originates in the plant site. Smaller amounts of Hg were used at other Oak Ridge facilities with similar results. Although the primary Hg discharges from Y-12 NSC stopped in 1963, small amounts of Hg continue to be released into the creek from point sources and diffuse contaminated soil and groundwater sources within Y-12 NSC. Mercury concentration in EFPC has decreased 85% from ∼2000 ng/L in the 1980s. In general, methylmercury concentrations in water and in fish have not declined in response to improvements in water quality and exhibit trends of increasing concentration in some cases.