Evaluation of trace element contamination using Armadillo officinalis Duméril, 1816 (Crustacea,Isopoda) as a tool: An ultrastructural study

To estimate trace element bioaccumulation in Armadillo officinalis, specimens were collected from Ghar El Melh lagoon then exposed for 3 weeks in contaminated sediments with copper, zinc, and cadmium. From the first week until the end of the experiment, a decrease in A. officinalis growth related to the increase of Cd concentration in the sediment was recorded. However, a mass gain was highlighted under Cu and Zn exposures. At the end of experiment, body metal concentrations were measured using flame atomic emission spectrometry. Results of the bioaccumulation factor showed that the species could be considered as a macroconcentrator of copper (BAF > 2) and a deconcentrator of zinc (BAF < 2). Microscopy observations of hepatopancreas cells showed morphological and histological changes even at the lowest concentration. They consisted in the microvillus border destruction, lipid droplets modifications, trace element accumulation, and the condensation of the majority of cellular organelles. The degree of these alterations was found to be dose‐dependent. Through these results, the isopod A. officinalis could be used as relevant monitor organisms for soil metal contamination.     

Physiological and histopathological responses of Porcellio laevis (Isopoda,Crustacea) as indicators of metal trace element contamination

This study was designed to assess the impact of the mixture of cadmium (Cd) and zinc (Zn) on the bioaccumulation and the ultrastructural changes in the hepatopancreas of Porcellio laevis (Latreille, 1804) after 4 weeks of exposure to contaminated Quercus leaves under laboratory conditions. For each metal, four concentrations were used with four replicates for each concentration. Metal concentrations in the hepatopancreas and the rest of the body were determined using atomic absorption spectrometry. From the first week until the end of the experiment, a weight gain in P. laevis was observed particularly between the first and the end of exposure from 93.3 ± 18.22 mg fw to 105.22 ± 16.16 mg fw and from 106.4 ± 22.67 mg fw to 125.9 ± 23.9 mg fw for Mix1 and Mix4, respectively. Additionally, the determined metal trace elements (MTE) concentrations in the hepatopancreas were considerably higher compared to those in the rest of the body and seem to be dose‐dependent. Using transmission electron microscopy (TEM), some alterations were highlighted in the hepatopancreas. The main observed alterations were (a) the destruction of the microvilli border in a considerable portion of cells, (b) the increase of the lipid droplets with different shapes and sizes, (c) the increase in the number of the mitochondria, and (d) the appearance of TE in the form of B‐type granules. The obtained results confirmed the ability of P. laevis to deal with high amounts of MTE, suggesting its possible use in future soil's biomonitoring programs.     

Trace elements bioaccumulation in Porcellionides pruinosus (Brandt, 1833) and related biomarkers of exposure

In the last years, the increase of anthropogenic activities has led to a progressive pollution of ecosystems. Oniscidean isopods are a group of terrestrial animals known as excellent bio‐indicators and bio‐accumulators, because of their capacity to accumulate contaminants and to implement appropriate strategies of detoxification. For this study, we collected Porcellionides pruinosus from two different sites in the Corinth region (Greece), a polluted site and a control site, to assess the heavy metal pollution status. We conducted an inductively coupled plasma mass spectrometry (ICP‐MS) analysis for the detection of trace elements in animals' hepatopancreas and soil; we performed also immunohistochemistry for detection of metallothioneins 1 (MT1) and heat shock protein 70 (HSP70s). The results suggest a great capacity of the species in trace elements bioaccumulation and the high degree of pollution of the area near Agioi Theodoroi where the metal pollution index (MPI) is threefolds higher than the control site, due to the presence of significantly higher concentration of arsenic, cadmium, cobalt, chromium, manganese, molybdenum, and lead found in hepatopancreas and soil. The environmental stress is confirmed by the expression of the relative biomarkers of exposure MT1 and HSP70. Our study, with a multimarker approach, consents getting a complete analysis of environmental quality and the effect of pollution on organisms and shows for the first time the heavy metals contamination status in certain Greece area. Furthermore, P. pruinosus proved to be an appropriate indicator of pollution in terrestrial ecosystems.     

The induced damage in the hepatopancreas of Orchestia species after exposure to a mixture of Cu/Zn—An ultrastructural study

The hepatopancreas of crustaceans species has been recognized as an essential target organ to assess trace elements' effects. Due to its dynamic and capability of detoxifying trace metal, this organ often indicates distinct pathological disturbances. In the present work, we intend to evaluate the bioaccumulation of trace metal in three Orchestia species (Orchestia montagui, Orchestia gammarellus, and Orchestia mediterranea) living in symmetry in the banks of Bizerte lagoon (37°13′8″N 09°55′1″E) after their exposure during 14 days to a mixture of copper and zinc, and to highlight the effect of these metals on their hepatopancreas ultrastructure using transmission electron microscopy. At the end of the experiment, results showed that the mortality and the body mass varied according to the used nominal concentrations. Significant alterations were noted in all the treatment groups. The degree of these alterations depends on the used concentration, and they are represented especially by the cells remoteness and the border lyses, the reduction of the nuclear volume, the increase in the cytoplasm density with the presence of trace metal in the nucleus as well as in the vacuole, the disorganization and the destruction of microvilli, the condensation of the majority of cellular organelles and mitochondria swelling. Through this study, Orchestia genus could be an attractive candidate for the biochemical study of trace metal toxicity in Tunisian wetlands.     

Morphostructural and immunohistochemical study for evaluation of nano‐TiO2 toxicity in Armadillo officinalis Duméril, 1816 (Crustacea,Isopoda, Oniscidea)

Invertebrates are precious organisms in order to study environmental pollution. In particular, they appear to be suitable as a bioindicator species for pioneer ecotoxicity studies on new xenobiotics such as nanoparticles. In fact, they are able to absorb nanomaterials scattered in the environment in different ways and it's known the compartmentalization of nano‐sized contaminants in selected tissues and intracellular organelles. Titanium dioxide represents the most used nanoparticulate, destined to become probably ubiquitous in the environment. Recently, some research has been published on the toxic potential of nano‐TiO₂ in several animal species. Among all invertebrates, Oniscidean Isopods are the only taxon of Crustaceans that has become completely terrestrial, known as excellent bioindicators and bioaccumulators. They have a digestive gland, the hepatopancreas, which is the location of election for the accumulation of pollutants. For this reason, they are considered efficient animal models to ecological studies. For this study, we collected Armadillo officinalis from Natural Oriented Reserve of “Vendicari” (Sicily, Italy), to evaluate the toxicity of titanium dioxide (TiO₂) on their hepatopancreas, after a short period of exposure. We conducted morphostructural and immunohistochemistry assays. The results suggested a great capacity of the species of bioaccumulation of nanoparticles in the hepatopancreas, where a strong positivity to the metallothioneins was highlighted. Our study confirms that Oniscidean Isopods, in particular Armadillo officinalis, proved to be an appropriate indicator of pollution in terrestrial ecosystems from nanoparticles.     

Chemi-emission of electrons from metal surfaces in the cutting process due to metal/gas interactions

The exo-electron emission behaviour from Ti, W, Mo, Fe, Al, Ni, Cd, Zn and Pb metals was investigated during and after being cut in O₂ and N₂ gases at a gas pressure of 3 × 10⁻² Pa and in a vacuum of 4 × 10⁻⁴ Pa. The results showed that the emission behaviour of electrons during and after cutting depends on the combination of metal and the surrounding gas species. The emission intensity of electrons while cutting various metals in O₂ gas increased sharply with an increase in the negative value of the heat cf formation, ΔH_f of the oxide. The emission intensity in N₂ was also higher at higher negative values of ΔH_f of the metal nitride. Electron emission intensity from the cut metal surface is concluded to be a function of the heat of formation of reaction products of the metal surface with the surrounding gas.     

Bioaccessibility of heavy metals in the seaweed Caulerpa racemosa var. corynephora: Human health risk from consumption

This study reports the total and bioaccessible concentrations of heavy metals (Mn, Fe, Cu, Zn, Cd and Pb) in the seaweed Caulerpa racemosa var. corynephora collected from local markets along the Andaman coast of Krabi Province, Thailand. Microwave-assisted acid digestion (EPA Method 3052) was used for sample preparation prior to total metal analysis. The in vitro Unified Bioaccessibility Method (UBM) was applied to assess the bioaccessibility of the metals in the seaweed samples. The total, gastric phase, and residual fraction concentration were determined by inductively coupled plasma-optical emission spectrometry (ICP-OES). The total amounts found in the seaweed samples, in ascending order were Cd?in vitro bioaccessibility method.     

The influence of water vapour in air on the friction behaviour of pure metals during fretting

An investigation was conducted to determine the effect of water vapour content in air on the frictional behaviour during fretting of pure metals: iron, aluminium, copper, silver, chromium, titanium and nickel. The fretting experiments were carried out under various humidity levels, ranging from dry air to 50% relative humidity at 23°C. During the experiment the frictional force between fretting surfaces was measured. Pure metals, except iron, were found to have a maximum value of the coefficient of friction during the steady-fretting stage (μ_s) at a specific humidity (RH_max). Iron showed a rapid decrease in μ_s with increasing humidity at RH_max. Each pure metal also exhibited maximum fretting wear at RH_max. The value of μ_s at RH_max for each metal was strongly related to the heat of formation of the lower metal oxide, indicating that the adhesive contact area was larger at RH_max for the fretting of metals with less chemical activity. At high humidity levels water vapour generally reduced the coefficient of friction, μ_s.     

Electron and ion imaging of gland cells using the FIB/SEM system

The FIB/SEM system was satisfactorily used for scanning ion (SIM) and scanning electron microscopy (SEM) of gland epithelial cells of a terrestrial isopod Porcellio scaber (Isopoda, Crustacea). The interior of cells was exposed by site-specific in situ focused ion beam (FIB) milling. Scanning ion (SI) imaging was an adequate substitution for scanning electron (SE) imaging when charging rendered SE imaging impossible. No significant differences in resolution between the SI and SE images were observed. The contrast on both the SI and SE images is a topographic. The consequences of SI imaging are, among others, introduction of Ga⁺ ions on/into the samples and destruction of the imaged surface. These two characteristics of SI imaging can be used advantageously. Introduction of Ga⁺ ions onto the specimen neutralizes the charge effect in the subsequent SE imaging. In addition, the destructive nature of SI imaging can be used as a tool for the gradual removal of the exposed layer of the imaged surface, uncovering the structures lying beneath. Alternative SEM and SIM in combination with site-specific in situ FIB sample sectioning made it possible to image the submicrometre structures of gland epithelium cells with reproducibility, repeatability and in the same range of magnifications as in transmission electron microscopy (TEM). At the present state of technology, ultrastructural elements imaged by the FIB/SEM system cannot be directly identified by comparison with TEM images.     

Physiological and behavioral responses of Orchestia gammarellus (Amphipoda,Talitridae) towards trace elements contamination soil

Effects of trace elements on the hepatopancreas ultrastructure and on the locomotor activity rhythm were investigated in the amphipod talitrid Orchestia gammarellus collected from Bizerte lagoon banks situated in the north of Tunisia. Animals were exposed to a series of contaminated soil with different concentrations of cadmium and zinc. The locomotor activity rhythm was studied under constant darkness. Histopathological analysis showed that the trace elements, especially the Cd, induced significant changes in the morphology and in the ultrastructural organization of hepatopancreatic cells. The significant alterations obtained were dose dependent. Concerning the behavioral response, results revealed the presence of two components whatever the experimental conditions. In addition, a great inter-individual variability of the locomotor rhythm was observed. Patterns were in majority bimodal for the control individuals and became unimodal and multimodal when exposed under Cd and Zn, respectively. Furthermore, ultradian and circadian periods were determined. The circadian period lengthened after Zn exposure. In addition, the locomotor activity rhythm was more stable for control individuals. However, those exposed to Cd were less active.     

Insight of EDX analysis and EFTEM: Are spherocrystals located in strombidae digestive gland implied in detoxification of trace metals?

Digestive tubules of Strombidae are composed by three cell types: digestive cells, vacuolated cells, and crypt cells. The last one is characterized by the presence of intracellular granules identified as spherocrystals. Such structures are known to occur in basophilic cells of gastropod digestive gland, where they are supposed to be involved in the regulation of some minerals and in detoxification. In this study, energy‐dispersive X‐ray analysis (EDX) and energy filtered transmission electron microscopy (EFTEM) were used to determine the elemental content of spherocrystals in two Strombidae, Strombus gigas and Strombus pugilis. In freshly collected individuals of both species, the following elements were detected: Ca, Fe, Mg, P, and Zn. Aluminum and Mn were also detected in S. gigas. Their presence in spherocrystals indicates that, in Strombidae, spherocrystals are involved in the regulation of minerals and essential trace metals. In order to answer the question “are spherocrystals involved in nonessential trace metals scavenging?,” artificial cadmium and lead exposure by both waterborne and dietary pathways was applied to S. pugilis. No evidence of cadmium (Cd(NO₃)₂) or lead (Pb(NO₃)₂) provided by food was found in spherocrystals. Cadmium provided in water (Cd(NO₃)₂ and CdCl₂) causes structural modifications of the digestive gland; however, this element was not trapped in spherocrystals. These results suggest that spherocrystals are not involved in detoxification of such nonessential trace metals. Microsc. Res. Tech., 2011. © 2011 Wiley‐Liss, Inc.     

Elemental changes in the brain, muscle, and gut cells of the housefly, Musca domestica, exposed to heavy metals

The toxic effects of heavy metals on organisms are well established. However, their specific action at the cellular level in different tissues is mostly unknown. We have used the housefly, Musca domestica, as a model organism to study the toxicity of four heavy metals: copper (Cu), zinc (Zn), cadmium (Cd), and lead (Pb). These have been fed to larvae at low and high, semi-lethal concentrations, and their accumulation in the head, thorax, and abdomen was subsequently measured in adult flies. In addition, their impact on the cellular concentration of several elements important for cell metabolism-sodium (Na+), magnesium (Mg++), phosphorous (P), sulphur (S), chloride (Cl-) and potassium (K+)-were measured in neural cells, muscle fibers, and midgut epithelial cells. Our study showed that the heavy metals accumulate mainly in the abdomen, in which the concentrations of two of the xenobiotic metals, Cd and Pb, were 213 and 23 times more concentrated, respectively, than in controls. All the heavy metals affected the cellular concentration of light elements in all cell types, but the changes observed were dependent on tissue type and were specific for each heavy metal, and its concentration.     

Numerical simulation of the localization behavior of hydrostatic-stress-sensitive metals

The present paper deals with the numerical simulation of the elastic–plastic deformation and localization behavior of solids which are plastically dilatant and sensitive to hydrostatic stresses. The model is based on a generalized macroscopic theory taking into account macroscopic as well as microscopic experimental data obtained from tests with iron-based metals. It shows that hydrostatic components may have a significant effect on the onset of localization and the associated deformation modes, and that they generally lead to a notable decrease in ductility. The continuum formulation relies on a generalized I₁–J₂–J₃ yield criterion to describe the effect of the hydrostatic stress on the plastic flow properties of metals. In contrast to classical theories of metal plasticity, the evolution of the plastic part of the strain rate tensor is determined by a non-associated flow rule based on a plastic potential function which is expressed in terms of stress invariants and kinematic parameters. Numerical simulations of the elastic–plastic deformation behavior of hydrostatic-stress-sensitive metals show the physical effects of the model parameters and also demonstrate the efficiency of the formulation. Their results are in excellent agreement with available experimental data. A variety of large-strain elastic–plastic problems involving pronounced localizations is presented, and the influence of various model parameters on the deformation and localization behavior of hydrostatic-stress-sensitive metals is discussed.     

MoS2‐based alloys and nanocomposites for solid lubrication

The development of MoS₂ coatings has involved the modification of substrate surfaces, the addition of metals or compounds to the MoS₂, and variation in the deposition process parameters affecting the properties of deposited films. More recently, multilayer and periodic nanolayer coating structures have also been investigated. At present, work is concentrated on alloys of MoS₂, mainly with various metals, and targeted at terrestrial (ambient air) applications. The addition of metals or compounds to physical‐vapour‐deposited MoS₂ has led to improvements in coating performance, for example, greater stability of friction coefficient, greater film endurance, and increased temperature/oxidation resistance. The metal or compound can be either in the form of nanoscale multilayers or mixed with the MoS₂, sometimes leading to nanoclusters within a MoS₂ matrix. Microstructural analysis seems to show that the primary function of these additives is to suppress the formation of low‐density, columnar structures. At certain concentrations an added metal can also enhance the formation of the tribologically favourable (002) orientation of the MoS₂ crystallites. Other changes in the properties of MoS₂—metal composites may be due to their oxidation resistance, as indicated by the stability of these films against storage in air and their increased endurance when in sliding contacts at elevated temperatures.     

双金属核壳纳米结构中荧光单分子的表面能量转移及金属操控自发辐射效应

贵金属表面附近的荧光分子因其表面等离子体共振的影响,荧光发射特性发生显著变化,被广泛应用在荧光探针等纳米器件的设计、开发中。荧光分子与金属之间的能量转移机制是设计此类荧光探针的基础。使用时域有限差分(FDTD)方法,对Au/SiO₂/Ag核壳纳米复合结构的等离子体杂化场中荧光单分子的表面能量转移(SET)效应和金属操控自发辐射效应进行了理论仿真研究。研究了金核和银壳共同作用时,荧光分子的SET和金属调控自发辐射过程随荧光分子位置及分子偶极矩取向的变化规律。计算结果表明,由于金核和银壳之间的局域表面等离子体共振杂化耦合,荧光分子与金属间的能量转移效率与距离d呈现出10次方的关系,这一结果明显区别于常规的荧光共振能量转移(FRET)效应,较之单金属结构的SET效应更加剧烈。这一结果有希望在生物光子学领域的纳米级局域光源的创建和生物分子的检测中得到应用。     

“Microscopic evidences of heavy metals distribution and anatomic alterations in breaching‐leaves of Cupressus lindleyi growing around mining wastes”

In this article a study of the distribution of heavy metals in Cupressus lindleyi breaching‐leaves was done in Taxco, Guerrero. At the same, heavy metals micro‐localization was conducted in the breaching‐leaves to understand the structural changes provoked by mining waste on plants. The most abundant contaminants in soils, tailings and different plant organs (roots, stems, and leaves) were Zn, Mn, and Pb. Nevertheless, As was more accumulated in the stem and breaching‐leaves. The translocation factor and the bio‐concentration factor were less than 1. The structural changes observed were the great accumulation of starch grains and phenolic compounds in the palisade parenchyma, changes in the hypodermis cell wall and necrotic zones in the palisade parenchyma. The distribution of heavy metals in breaching‐leaves tissues was homogeneous in most of the elements. These results showed that C. lindleyi is a species that can be employed in phytostabilization of contaminated zones with mining waste because it is a native plant that does not require a lot of conditions for its development. Microsc. Res. Tech. 77:714–726, 2014. © 2014 Wiley Periodicals, Inc.     

Study of plowing and friction at the surfaces of plastic deformed metals

Experimental and analytical investigations of plowing and friction were conducted at the surfaces of well-polished lead, aluminum, copper, nickel, molybdenum, and tungsten to study the mechanism of the load/penetration dependency. The experimental tests were performed with a Nano-Indenter XP of MTS and a Scanning Probe Microscope (SPM), Nanoscope IIIa of Digital Instruments. In addition to make indentation and measure the hardness and Young’s modulus, the indenter was used to make scratches at the surface of metals under different normal load while the penetration depth and frictional force encountered during the scratching were recorded. The SPM, operated mostly in the contact mode, was used to examine the scratch profile. Under the test conditions, plastic deformation dominated at the surfaces of the metals. An analytical model was established to express plastically deformed contacts, based on plowing of a conical-shaped indenter with a hemispherical tip at a plastic deformed surface. Penetration depth and scratched volume were calculated, which is in good agreement with experimental observation. The frictional coefficient μ was also calculated with the model, which accounted for plowing as well as the adhesion force between the indenter and surface. Beside fair agreement of experimental data and calculated values on μ under the loads applied, the model indicated a dramatic rise in friction coefficient under very low loads, which was not observed in the tests. The discrepancy was discussed, and it was believed that the dramatic increase in μ is for the calculated μ and may be due to the assumed dominant contribution of adhesion force in actual contact load with decreasing external load, and it appears only the adhesion energy Δγ is significant. The actual adhesion energy Δγ between our diamond indenter and metal surface in our test condition might be smaller than the value used in calculation.     

Response of <i>MaHMA2</i> gene expression and stress tolerance to zinc stress in mulberry (<i>Morus alba</i> L.)

HMA2 (heavy metal ATPase 2) plays a crucial role in extracellular and intracellular Zn²⁺ transport across biomembranes, maintaining ion homeostasis, and playing an important role in the normal physiological metabolism, growth, and development of plants. In our study, a novel HMA2 gene, named MaHMA2, was isolated and cloned from white mulberry (Morus alba L.). The gene sequence obtained was 1,342 bp long, with an open reading frame of 1,194 bp, encoding a protein of 397 amino acids, with a predicted molecular mass of 42.852 kD and an isoelectric point of 7.53. This protein belonged to the PIB-type ATPase transport protein family. We analyzed the expression of the MaHMA2 gene by quantitative real-time PCR. The results showed that the level of MaHMA2 gene expression decreased to a Zn concentration of 800 mg/kg. Malondialdehyde and proline levels increased and responded to increasing Zn when the MaHMA2 gene was silenced, whereas the activities of peroxidase and superoxide dismutase tended to increase in response to increasing Zn²⁺ ion stress concentrations but were lower in the gene-silenced plants. These findings suggested that the MaHMA2 gene played an active role in the tolerance response of mulberry to Zn stress.     

The surface chemistry of vapor phase lubricants: tricresylphosphate on Ni(1 0 0)

Tricresylphosphate (TCP) is known to serve as an excellent high temperature vapor phase lubricant with some metals such as Fe but not with others such as Ni. The surface chemistry of m-TCP has been studied on clean and phosphorous covered Ni(1 0 0) surfaces in order to understand the differences between its reactivity on Fe and Ni. Our results show that upon heating to 800 K m-TCP decomposes on the clean Ni(1 0 0) surface to deposit carbon and phosphorous with the evolution of H₂, CO, benzene, and toluene into the gas phase. During further heating to 1000 K, all the carbon on the surface dissolves into the Ni bulk leaving only phosphorous. The adsorption and heating of m-TCP on the phosphorous-modified Ni surface does not result in significant decomposition. The clean Ni substrate is able to activate TCP decomposition in much the same way that it decomposes on the clean Fe surface. The lack of significant differences in the chemistry of TCP on the clean metal surfaces cannot explain the fact that TCP vapor will form thick lubricating films on Fe but not on Ni.     

The effect of different pollutants exposure on the pigment content of pigmented macrophage aggregates in the spleen of Vardar chub (Squalius vardarensis Karaman, 1928)

Pigmented macrophage aggregates (MAs) are known to change under influence of various factors, such as aging, season, starvation, and/or pollution. In this study, changes in the pigment content of the MAs in the spleen of Vardar chub (Squalius vardarensis, Karaman) (n = 129) collected in spring and autumn, from three rivers with different pollution impact was examined: Zletovska River (metals), Kriva River (metals and municipal wastewater), and Bregalnica River (municipal wastewater). Collected data revealed increased relative volume and number of MAs containing hemosiderin under the influence of metals, significant in autumn (p < .05). In chub exposed to metals combined with municipal wastewater, significant increase of lipochrome accumulation in MAs in autumn, melanin in MAs in fish captured in the spring season, and number of splenic MAs containing combination of melanin and lipochrome was noted. Volumes and number of MAs containing combination of hemosiderin and lipochrome increased in spleen of fish captured in autumn from both Zletovska River and Kriva River, most likely due to the contribution of hemosiderin and lipochrome, respectively. Values measured for the various pigments in splenic MAs in fish captured from Bregalnica River, were overall closer to the values measured for fish captured from Kriva River. Notably, melanin and lipochrome are more likely to be found in fish from waters influenced by municipal wastewater (organic pollution) and hemosiderin in fish spleen from water influenced by mining activity (heavy metals pollution).