Methodological aspects of moss sample preparation. Effects of freezing and duration of washing on the cellular distribution of elements in Fontinalis squamosa Hedw.

Aquatic bryophytes are widely used in biomonitoring studies, mainly because of their capacity to act as bioaccumulators. As different methods can be used to preserve and process moss samples, the results of elemental analyses may also vary, particularly if the contents of different cellular compartments are analyzed separately. In the present study, we compared the total concentrations of some elements and the concentrations of these in different cellular locations in frozen-thawed and fresh samples of the aquatic moss Fontinalis squamosa that were also washed for different lengths of time before analysis. We used the sequential elution technique (SET) to extract the different fractions, and we determined the concentrations of K, Mg, Ca, Cd, Co, Cu, Al and Zn in the extracts. The results varied depending on the element, cellular location and moss sampling site. In the moss samples processed after freezing, the greatest differences were in the intracellular concentrations of K, Mg and Cd, which were lower than in the fresh samples. Minor differences were found in the concentrations of elements in other cellular locations and in the total concentrations of elements. The increase in the duration of the initial washing step, carried out to remove soluble and particulate intercellular elements, may also cause the release of elements (e.g. K and Mg) bound to extracellular exchange sites. The concentrations in the other cellular fractions and the total concentrations were less affected by the washing duration. Neither freezing nor the use of long washing times are recommended for processing moss samples prior to extraction of elements by the SET. Further studies are needed to confirm and clarify the observations.     

Intracellular and Extracellular Uptake of Cadmium by the Moss Rhytidiadelphus squarrosus

Intra- and extracellular Cd uptake were investigated in themoss Rhytidiadelphus squarrosus. Intracellular Cd uptake displayedMichaelis–Menten kinetics and showed that the moss hada high uptake rate and high affinity for Cd. Extracellular Cdbinding capacity was also high. The anion used had little effecton Cd uptake to either location but both intra- and extracellularCd uptake were reduced by equimolar concentrations of Ca, Mgor Zn, although inhibition was not wholly competitive. IntracellularCd uptake was not significantly affected by the supply of energygenerating reserves but, like extracellular Cd uptake, showedaltered characteristics following growth under controlled laboratoryconditions. The results are compared with data on Cd uptakeby the lichen genus Peltigera. Rhytidiadelphus squarrosus, moss, cadmium, intra- and extracellular uptake characteristics, kinetics     

The impact of bacterial size on their survival in the presence of cationic particles of nano-silver

BackgroundMicrobial surface area is one of the battlegrounds for invading microbes and host defense. Hence, infectious diseases caused by drug resistant microbes with large surface area are more difficult to treat than small size microbes. Nanobiology offers opportunities to re-explore the biological properties of conventional drugs at molecular level to combat these microbes. The purpose of the present study was to examine size depended susceptibility of Gram-positive bacteria towards nano-silver particles.MethodsThis study investigated the growth, surface charge, and morphology of emerging B. megaterium MTCC 7192 and re-emerging S. aureus MTCC 3160 cells in order to observe the susceptibility of these bacteria towards cationic nano-silver particles. Nano-silver particles were applied into wells formed on the Nutrient agar plates containing 10⁸ CFU/mL of the bacteria. Surface potential of normal and treated cells was measured by Microtrac and the effects of nano-silver particles on bacterial cells were assessed by Scanning Electron Microscopy (SEM).ResultsIn this work, synthesized nano-silver particles were found to be more effective against B. megaterium MTCC 7192 than S. aureus MTCC 3160. For B. megaterium MTCC 7192, a 0.30 fold increase in inhibition zone was observed after the addition of nano-silver particles in the wells. From our studies, it is reasonable to state that alternation of zeta potential may affect the cell morphology, which was further confirmed by SEM.ConclusionThe present study concluded that nano-silver particles appears to interact with a larger surface area more effectively.     

Accumulation of Pb and Zn in Gametophytes and Sporophytes of the Moss Funaria hygrometrica(Funariales)

Accumulation of lead and zinc was studied in the moss Funariahygrometrica Hedw. collected from mine tailings. Heavy metalaccumulation in gametophytes and sporophytes was quantifiedby graphite furnace atomic absorption spectrometry (GFAAS) andinductively coupled plasma-atomic emission spectrometry (ICP-AES).Pb and Zn accumulation in the placental zone was analysed byx-ray scanning electron microscopy (SEM) and transmission electronmicroscopy (TEM) microanalysis. Spectrometry showed that whilemoss gametophytes accumulated considerable concentrations ofheavy metals, sporophytes accumulated only small concentrationsof metals. X-ray SEM and TEM showed that the two metals accumulatedin placental transfer cells on both the gametophytic and sporophyticsides. To investigate the uptake pattern for both metals undercontrolled conditions, F. hygrometrica plants collected froma non-polluted site were treated in the laboratory with separatesolutions of Pb and Zn at two concentrations (10^-2and 10^-4 M)for 24 or 168 h. Metal accumulation was analysed separatelyin gametophytes and sporophytes using GFAAS and ICP–AES.Each generation had a different accumulation quotient for bothmetals, and gametophytes accumulated significantly more metalthan sporophytes. Concentrations of Zn in sporophytes were alwayshigher than concentrations of Pb. The findings are discussedin relation to the role performed by the gametophyte and theplacenta in the accumulation and sequestration of Pb and Zn.Copyright 2001 Annals of Botany Company Atomic spectroscopy, Funaria hygrometrica, gametophyte, Pb and Zn accumulation, sporophyte, x-ray TEM and SEM microanalysis     

Biochemical responses of the aquatic moss Fontinalis antipyretica to Cd,Cu, Pb and Zn determined by chlorophyll fluorescence and protein levels

Biochemical reactions to Cu, Cd, Zn and Pb in the aquatic moss Fontinalis antipyretica were studied in order to characterize the physiological background of the metal response. Chlorophyll fluorescence and intracellular metal localization and stress protein levels were measured. Exposure to 25 or 100 μM Cu over a 7-day period resulted in a decline of chlorophyll fluorescence to about 70% and 52%, respectively. Up to 100 μM Cd caused a decrease in chlorophyll fluorescence to 75%. With all metals used at 25–100 μM concentrations, the intracellular uptake increased. For all metals investigated at 25–100 μM, the intracellular uptake increased. Maximum values were reached at 100 μM Cu, Pb, Zn or Cd exposure. As shown by analytical electron microscopy (EDX, EELS) moss material treated with 50 μM Cu exhibited reduced sulphur levels in the cytoplasm and an increase in phosphate in vacuolar dense particles. EEL-spectra indicated that Cu is chelated in the cytoplasm by SH-groups and coprecipitated with orthophosphate in vacuoles. To monitor the stress response at the protein level, heavy metal induced heat shock protein 70 (hsp70) was measured. An antibody was raised against conserved plant metallothionein p2 motifs derived from Brassica juncea. In all metal-treated samples the antibody bound to proteins of about 8 kDa. However, sequencing failed to reveal significant homologies to known proteins. These experiments provide for the first time results on protein level after heavy metal stress in the aquatic bioindicator moss.     

Chromosome studies on some British and Irish mosses. I

Abstract

An electron microscope study of experimental and environmental lead poisoning of leaf cells from the moss Rhytidiadelphus squarrosus (Hedw.) Warnst. is described. Plants treated with lead acetate and lead chloride solutions, or collected from roadsides exposed to lead pollution from motor exhaust gases have been used. The results are compared with other studies of experimental lead poisoning in both plants and animals, with particular regard to uptake mechanisms and to lead accumulating at different sites in the cell. We have shown that lead may be bound within the cell wall, and that it may also enter the cytoplasm. Pinocytosis was observed in leaf cells from moss plants treated with lead salt solutions. Via pinocytotic vesicles within the cytoplasm the particles containing lead could then be discharged into the vacuole. In the leaf cells of mosses exposed to environmental pollution the protoplasmic lead was found to be bound within the nucleus, and was easily identified as electron-dense, nuclear inclusions.     

Antibody coated gold particles containing radioactive gold in the demonstration of cell surface molecules

Summary Gold particles of varying size which contain either ¹⁹⁵Au or ¹⁹⁸Au were prepared using white phosphorus or sodium citrate as the reducting agent. After coating with specific antibody to blood group A antigen or human IgG these particles were used to determine the number of particles binding to the surface of A₁ RBC's or rat RBC's to which human IgG had been attached. The number of particles binding to the surface of cells correlated with the number of antibody coated gold particles in the fluid bathing the cells as well as the number of antigen molecules on the cell surface. That is, the number of particles binding increased as the particle density of the suspension increased and as the cell surface antigen density increased. Under the conditions of the experiments, both blood group A antigen and human IgG appeared to be randomly distributed over the surface of the cells in TEM and SEM preparations. This approach permitted the quantitation of the number of gold particles bound per cell and at the same time, the examination of the distribution of the particles over the surface of the same cell population by TEM and SEM.     

An adaptative pattern for water conduction in the ectohydric moss Zygodon viridissimus var. rupestris Hartm.

Abstract

A detailed LM, SEM and TEM study of the dorsal and ventral surfaces of the leaf of the ectohydric moss Zygodon viridissimus var. rupestris (Z. baumgartneri) is described. The presence of papillae on both surfaces is confirmed and a groove, running the length of the nerve, is demonstrated. The presence of waxy particles on both surfaces is also demonstrated. A pattern for water conduction which could represent another interesting example of eco-physiological adaptation in bryophytes, is suggested.     

Amorphous mineral incrustations in the moss Homalothecium sericeum

Abstract

Internal mineral deposits in stem and seta were found in living, healthy-looking specimens of the moss Homalothecium sericeum from a Spanish locality. The deposits consisted of an amorphous crust of smooth texture. After examination with EDS probes coupled to a transmission and a scanning electron microscope, and X-ray diffraction analysis, the minerals were identified as mainly opal, carbonates, and Fe and Al hydroxides, although it is irregular throughout a given mineralized region of the moss. These mineral crusts are considered to be the product of biomineralization of ions taken from the substratum. A comparison is established between these deposits and the mineral crystals found in a previous study of Hookeria lucens.     

The use of the three-ridge clam (Amblema perplicata) to monitor trace metal contamination

The three-ridge clamAmblema perplicata was used to monitor two streams for the presence of zinc (Zn) and cadmium (Cd) derived from an industrial source. Clams were collected from a relatively uncontaminated area in one river and transported to four study sites in the two contaminated streams. The clams were placed into polyethylene cages and left in these streams for one week. Control clams were treated in a similar manner and left in the uncontaminated river.The highest mean concentration of Zn (956 µg/g dry wt) was found in the gill tissue of clams from the most contaminated site. The highest mean Cd concentration (18.6 µg/ g dry wt) was found in digestive glands of clams, also from the most contaminated site. Mean concentrations at contaminated sites were significantly higher than background and control levels; the findings suggest that these particular organs of clams may be useful in monitoring levels of Zn and Cd in other freshwater systems.     

Sanionia uncinata (Hedw.) loeske as bioindicator of metal pollution in polar regions

The length of gametophytes in the moss Sanionia uncinata and concentrations of the elements Cd, Co, Cr, Cu, Fe, Mn, Na, Ni, Pb, V and Zn in this moss and in the parent rock material were measured in West Spitsbergen (Svalbard). Samples were collected at different distances from the seashore from pure colonies in a wet moss tundra, a moderately wet moss and herb tundra, and a dry rock and terrestrial tundra. Not any statistical relation (PCCA) between concentration of elements in mosses and type of tundra habitat could be found. The principal component and classification analysis (PCCA) ordination revealed that S. uncinata from sites the most close, the most remote and on an intermediary distance from the seashore differentiated by the value of factor 1, which relates negatively to concentrations of Cd, Co, Cr, Cu, Fe, Pb and V. S. uncinata from sites situated the most close to and the most distant from the seashore was differentiated by the value of factor 2, which was negatively related to concentrations of Na, Ni and Mn in this moss. The established model points that Na, Mn, Cu, Ni and Zn were accumulated by S. uncinata mostly from sea spray.     

Cadmium Tolerance in a Metal-contaminated Population of the Grassland Moss Rhytidiadelphus squarrosus

Two populations of Rhytidiadelphus squarrosus, from metal-contaminated and uncontaminated habitats, differed in their intra- and extracellular Cd contents but had similar cellular levels of Cu. Moss shoots were supplied with a pulse of toxic metal by incubating in Cu or Cd nitrate solutions and effects on respiration, photosynthesis and intracellular K loss were monitored with time after initial exposure. Increasing intracellular Cu levels correlated most closely with a concurrent decline in intracellular K. Photosynthesis also declined in proportion to intracellular Cu; significant Cu-induced stimulation of respiration was observed. The most significant effect of Cd treatment was a decline in photosynthesis in proportion to the intracellular concentration of Cd. Apical segments from both populations showed similar sensitivity to Cu, whereas the metal-contaminated population showed increased resistance to Cd. Sensitivity to Cd increased in the more basal portions of moss gametophores, indicating that apparent resistance of Cd might reflect shoot vitality and age effects. After laboratory growth to eliminate differences in the physiological status of apical segments, it was confirmed that the metal-contaminated population of the moss was photosynthetically more tolerant to Cd at intracellular Cd concentrations found to cause considerable photosynthetic inhibition in the uncontaminated population. The metal-contaminated population of the moss that was tolerant to Cd was not co-tolerant to Cu.     

Survey of atmospheric heavy metal deposition in Suqian using moss contamination

Abstract

Heavy metal pollution in the atmosphere is a major environmental problem, which has important impacts on ecosystems and human health. In this study, the atmospheric deposition of cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni), lead (Pb), vanadium (V), and zinc (Zn) in Suqian, China, was investigated by using the moss Haplocladium microphyllum as a bioindicator. The survey was carried out during the summer of 2017 at 40 homogenously distributed sampling sites. Cr, Cu, Ni, Pb, V, and Zn were analyzed by inductively coupled plasma atomic emission spectrometry, and Cd was analyzed by inductively coupled plasma mass spectrometry. Wide variations in heavy metals (Cd, Pb, and Zn) indicated that the concentrations of elements are influenced by local emission sources. Spatial distribution maps of the elements were constructed using geographic information system technology. The pollution load index showed that the study area was moderately polluted to unpolluted. Significant positive correlations (p?<?.01) and weak positive correlations (p?<?.05) were identified among some of the elements, indicating that they originated from common sources. A principal component analysis classified the heavy metals into natural and anthropogenic sources, and identified four primary sources: nature soil dust, industry activities, traffic emission, and agricultural activities.     

Major and trace elements in Sphagnum moss from four southern German bogs,and comparison with available moss monitoring data

In this paper, we present concentrations of an array of major and trace elements (Ag, Al, As, Ba, Bi, Cd, Co, Cr, Cu, Fe, Mn, Mo, Rb, Sb, Sc, Sr, Th, Tl, U, V, Zn) in living Sphagnum mosses from four southern German bogs and compare them with moss monitoring data of the respective regions. To do this, Sphagnum mosses were collected in Upper Bavaria (Oberbayern, OB) and the Northern Black Forest (Nordschwarzwald, NBF). Surfaces of Sphagnum carpets were marked with plastic mesh and, one year later, the annual moss production was harvested. Up to 12 samples (40 cm × 40 cm) were collected per site, and 6–10 sites investigated per bog. The concentrations of these elements were then determined in acid digests using sector field ICP-MS. Variations within a given sampling site were in the range of 2 to 3-fold for all major and trace element concentrations except for Mn (12-fold) and Tl (38-fold). For most of the elements, concentrations between bogs of a given region were significantly different and atmospheric deposition of particles seems to be considerably affected by local circumstances such as tree canopy interception and microtopography. Comparing trace element concentrations measured in Sphagnum mosses for 2007 with published moss monitoring data for 2005 resulted in a very good agreement for most elements. Clearly, Sphagnum mosses from bogs are useful biomonitors for estimating atmospheric contamination by metals. This supports the use of Sphagnum in atmospheric deposition monitoring especially in cases where Sphagnum is abundant (e.g., boreal forests). In regions with neither bogs nor forests, living Sphagnum moss bags could be used to the same effect.     

METAL UPTAKE AND ALLOCATION IN TREES GROWN ON CONTAMINATED LAND: IMPLICATIONS FOR BIOMASS PRODUCTION

Phytostabilization aims to reduce environmental and health risks arising from contaminated soil. To be economically attractive, plants used for phytostabilization should produce valuable biomass. This study investigated the biomass production and metal allocation to foliage and wood of willow (Salix viminalis L.), poplar (Populus monviso), birch (Betula pendula), and oak (Quercus robur) on five different soils contaminated with trace elements (TE), with varying high concentrations of Cu, Zn, Cd, and Pb as well as an uncontaminated control soil. In the treatment soils, the biomass was reduced in all species except oak. There was a significant negative correlation between biomass and foliar Cd and Zn concentrations, reaching up to 15 mg Cd kg^?1 and 2000 mg Zn kg ^?1 in willow leaves. Lead was the only TE with higher wood than foliage concentrations. The highest Pb accumulation occurred in birch with up to 135 mg kg ^?1 in wood and 78 mg kg ^?1 in foliage. Birch could be suitable for phytostabilization of soils with high Cd and Zn but low Pb concentrations, while poplars and willows could be used to stabilise soils with high Cu and Pb and low Zn and Cd concentrations.     

Physiological Effects of Heavy Metals on the Moss Rhytidiadelphus squarrosus

Experiments on the effect of Cd, Cu, Ni, Pb and Zn, suppliedas single salt solutions between 10 µM and 0·1M, on the moss Rhytidiadelphus squarrosus showed little alterationto respiratory rates but reduced photosynthetic rates, and somemembrane damage as assessed by K leakage. Two distinct morphologicalforms of the moss showed different responses to supplied heavymetal. Storage of material, after 30 mins exposure to heavymetals, resulted in a further decrease in the photosyntheticrate. Expressing the photosynthetic decline relative to thetotal heavy metal recovered from the moss showed a similar pattern.Transfer of metal from extracellular exchange sites into theprotoplast was also demonstrated with storage after exposure.An approximately linear relationship was demonstrated betweenphotosynthetic decline and intracellular heavy-metal concentration,irrespective of the duration of exposure or morphological natureof the material used. Photosynthetic decline on storage is concludedto be a response to additional metal stress rather than a progressivedeterioration of the physiological process. Cadmium, copper, lead, nickel, zinc, photosynthesis, respiration, membrane damage, intracellular metals, Rhytidiadelphus squarrosus (Hedw.) Warnst     

Spatial distribution and environmental risk assessment of heavy metals identified in soil of a decommissioned uranium mining area

Abstract

The spatial distribution of six heavy metals (Cd, As, Zn, Pb, Cr, and Cu) in the soil of a decommissioned uranium mining area was investigated and their potential environmental risk was assessed. Soil samples were collected along the main riversides enclosing the mining area. The heavy metal distribution was determined by geospatial interpolation. Pearson correlation coefficient and principal component analysis were used to locate the sources of pollution which are the mine ore as natural source, and dressing plants and tailing area from human activity. The results indicate that the average concentrations of As and Cd strongly exceed the recommended EQSS (Environmental Quality Standard for Soils of China) limits at all sampling sites, whereas Zn concentrations were found to be slightly over the limit only at sampling sites close to the mining area. The concentrations of Cr, Cu and Pb were all within the recommended limits. Environmental risk was assessed using the toxicity characteristic leaching procedure defining the degree to which extend the metal is released into the solution. High leaching rates were found only for Zn and Cd, suggesting that together with its high concentrations Cd is the most toxic metal around the mining area, followed by As.     

Cadmium contamination in various environmental materials in an industrial area,Hangzhou, China

Abstract

Due to the location of manufacturing in north Hangzhou, many residential areas are close to trunk roads and factory areas. This potentially exposes citizens to pollution sources such as industrial emissions and vehicle exhaust. Because pollution by cadmium (Cd) is of great concern, an extensive survey was conducted in north Hangzhou city to study the current condition of Cd contamination and any resulting hazard. Environmental materials including water, bottom mud, urban green-land soil, street dust and moss were collected. Cd concentration and chemical partitioning in the samples were determined. The results indicate that all the media in north Hangzhou have elevated Cd concentrations, the Cd conten trend being: factory area >traffic area > residential area. The chemical partitioning results show that Cc in topsoil and street dust presents are mobile (9–61% Cd are bound in acetic acid extractable phase) which suggests a high ecological hazard potential. Environmental bioindicator moss samples were found to be loaded with Cd revealing high bioavailability of Cd in various media in north Hangzhou This confirms the possibility of Cd being a potential biohazard in this location.     

Insight into the pattern of heavy-metal accumulation in lichen thalli

BackgroundHeavy metals that pass through the plasmalemma are expected to influence on lichen metabolic processes; however, lichens may tolerate high concentrations of metals by sequestrating them extracellularly. Heavy metal accumulation level fundamentally determine the success of lichens in the colonisation of polluted sites; however, the proportions between extra- and intracellular metal concentrations in lichen thalli are still poorly recognized. In this study metal accumulation patterns of selected toxic trace elements, i.e. Pb, Cd, and micronutrients, i.e. Zn, Cu and Ni, in Cladonia cariosa thalli were recognised in relation to extra- and intracellular fractions.MethodsThe intracellular and total concentrations of Zn, Pb, Cd, Cu and Ni in lichen thalli collected from eleven variously polluted sites were determined by means of atomic absorption spectrometry. Additionally, organic carbon and total nitrogen contents as well as pH of soil substrate were measured.ResultsThe accumulation patterns differed between studied metal elements; the major part of Zn, Pb and Cd loads was accumulated extracellularly, whereas Cu and Ni accumulation was mostly intracellular. Like toxic trace elements, Zn was accumulated mainly extracellularly at high polluted sites. The non-linear models most reliably reflect relationships between intracellular and extracellular metal contents in C. cariosa thalli. The intracellular contents of Zn, Pb, Cd and Cu increased slower at higher than at lower extracellular concentrations. Moreover, at higher total concentrations of elements in the thalli, their extracellular proportions were markedly increased.ConclusionThe results suggest that in the face of extreme Zn-enrichment, lichens demonstrate the ability to accumulate the excess of Zn outside the cells. Therefore, it can be concluded that metal accumulation depend not only on the element but also on its abundance in the environment and direct availability for lichens. The studied species showed a defence against excessive intracellular accumulation when a given element is in excess. Such capability may facilitate the colonization of extremely polluted sites by certain pioneer lichens.     

Bioconcentration of Cd and Zn in the soils of an uncontaminated forest in the Quebec Laurentians

Fine-scale trace element (TE) concentration patterns exist in soils that were never exposed to the direct deposition of TE in the past. We aimed to understand the role of vegetation, in particular trees, in defining these spatial patterns in the soil horizons of an uncontaminated forest ecosystem. Living tree leaves and surface soil samples (L, F, and B horizons) were collected from a ca. 1400 m² plot in the Hermine watershed where a strong gradient in the abundance of Yellow birches (Betula alleghaniensis—BA) exists. TE analysis showed that BA leaves had up to 13 times more Cd and Zn than the other tree species. Significantly similar spatial patterns were detected between the density of BA trunks, the abundance of BA leaves in the litter (L) layer, and Cd and Zn concentrations in the F horizon. The opposite trend was found in the upper B horizon, where Cd and Zn were depleted under a denser BA canopy. We submit that soil depletion could be associated with the preferential phytoextraction of Cd and Zn by BA over a period of several decades. Our study stresses the role of biological cycling by trees on the profile redistribution and the generation of patterns of soil TE in quasi-pristine forests.