Field validation of a battery of biomarkers to assess sediment quality in Spanish ports

Two marine invertebrates, the crab Carcinus maenas and the clam Ruditapes philippinarum, were used as bioindicator species to assess contamination when exposed in situ to sediment from different sites from four Spanish ports Cadiz (SW Spain), Huelva (SW Spain), Bilbao (NE Spain) and Pasajes (NE Spain). In an attempt to determine sediments toxicity, a combination of exposure biomarkers was analyzed in both species: metallothionein-like-proteins (MTLPs), ethoxyresorufin O-deethylase (EROD), glutathione S-transferase activity (GST), glutathione peroxidase (GPX) and glutathione reductase (GR). In parallel, physical and chemical characterization of the different sediments was performed and biological responses related to the contaminants. Significant induction of MTLPs was observed when organisms were exposed to metal contaminated sediments (port of Huelva), and EROD and GPX activities after exposure to sediments containing organic compounds (port of Bilbao and Pasajes). No significant interspecies differences were observed in biomarker responses except for the GST and GR.     

Heavy metal bioavailability and effects: II. Histopathology-bioaccumulation relationships caused by mining activities in the Gulf of Cádiz (SW, Spain)

The relationship between bioaccumulation of heavy metals (Zn, Cd, Pb and Cu) and histological lesions in different tissues of organisms is assessed in three different areas located in the southwest of Spain in the Gulf of Cádiz (Ría of Huelva, Guadalquivir estuary and Bay of Cádiz) affected and non-affected by mining activities. Data included in these relationships were obtained along the years 2000 and 2001 to address the impact of the Aznalcóllar mining spill on the Guadalquivir estuary. The bioaccumulation and the histological lesions measured in this seasonal study in the Gudalquivir estuary were linked to derive tissue quality guidelines (TQGs) by means of a multivariate analysis approach (MAA). Sediments collected in the same areas of study were used to expose organisms during the survey carried out in autumn 2001 and to address the relationship between bioaccumulation and histological lesions under laboratory conditions and related to chemicals bound to sediments. Lesions show that the organisms collected in the ría of Huelva and exposed to their sediments were severe, intermediate in the Guadalquivir estuary and absent in the Bay of Cádiz. Results show that the Guadalquivir estuary trends to recover its initial status quo previous to the mining spill. The link between chemical concentration and the lesions measured in the same tissues using MAA permits to derive tissue quality guidelines for two organisms, oysters (Crassostrea angulata) and clams (Scrobicularia plana) collected in the Guadalquivir estuary and associated with the heavy metals from the mining spill (Zn and Cd). The TQG values expressed as concentrations (mgkg-1--dry weight) not associated with biological effects are for oysters, Zn, 8603, Cd, 3.42; and for clams Zn, 800, Cd, 2.6.     

Heavy metal bioavailability and effects: I. Bioaccumulation caused by mining activities in the Gulf of Cádiz (SW, Spain)

The bioaccumulation of six metals (Fe, Mn, Zn, Cd, Pb and Cu) was studied as part of the monitoring of the Aznalcóllar mining spill (April 1998) on the Guadalquivir estuary and in other estuaries located in the Gulf of Cádiz. Fish, clams and oysters were collected during different seasonal periods along the years 2000 and 2001 in the Guadalquivir estuary to determine the bioaccumulation of the metals originated by the mining spill. Results were compared to the bioaccumulation of the same metals in fish and clams exposed in the laboratory to sediments collected in the same areas during autumn 2001. The bioaccumulation of these metals was compared to the concentration of metals measured in tissues of same taxas collected in the areas of the ria of Huelva and the Bay of Cádiz. Results show that the bioaccumulation of Zn and Cd in the organisms sampled in the Guadalquivir estuary was associated with the enrichment of these metals in the estuary from the mining spill and decreased along the time reaching the lowest values in autumn 2001. The metal Cu show different trends that are associated with other sources of contamination than the spill and related to the transport of this metal from Huelva to Guadalquivir estuary and/or to the use of this metal as plaguicide in the rice fields located in the area. The comparison between bioaccumulation results under field and laboratory conditions obtained in the different areas of study shows that these data can be used to discriminate between acute and chronic impacts associated with mining activities.     

Heavy metal concentrations in molluscs from the Atlantic coast of southern Spain

Trace metals were determined in the two most abundant species of bivalve molluscs along the Atlantic coast of southern Spain (Donax trunculus and Chamelea gallina) and in the sediments where they live. The results show that the area near the mouth of the Huelva estuary is where the highest metal concentrations are found in sediments and in the two bivalve species. This is not surprising, considering that the Huelva estuary is the mouth of the Tinto and Odiel rivers, which have one of the highest levels of metal pollution of all the rivers of Europe. The two species of bivalves have different amounts of metals in their tissues. The concentrations of Cr, Cu, Pb, Zn, As and Hg in D. trunculus were significantly higher (p < 0.05) than in C. gallina; however, C. gallina contained more Ni and Cd. In both species the most abundant elements were Cu and Zn, while Hg showed the lowest values. There is a significant correlation (p < 0.05) for concentrations of Cu, Pb, Zn and Hg in D. trunculus and C. gallina relative to their concentrations in surface sediments.     

Trace element contamination of Norwegian Lake sediments

Concentrations of Sb, Hg, Bi, Cd, Mo, As, Co, Ni, Cr, Cu, V, Pb and Zn in surface and preindustrial sediments from 210 lakes in Norway were used for studying modern atmospheric depositions of these elements. Surface sediments had considerably higher concentrations of Sb, Hg, Bi, Cd, As, Pb than preindustrial sediments. The differences decreased with latitude and altitude. A multivariate analysis including the trace elements and the major constituents (organic matter, Si, Al, Fe and Mn) of surface sediments suggested the following relationships: Sb, Hg, Bi, As, and Pb formed a group with strong associations to organic matter. Ni, Cr and Cu formed a second group, weakly associated to the inorganic sediment fraction (Si and Al). Zn and Cd formed a third group with weak associations to organic matter. Co was associated to Mn, whereas Mo and V showed no important covariations with any other trace elements or major components.     

Assessment of trace element pollution and ecological risks in a river basin impacted by mining in Colombia

Trace element pollution in rivers by anthropogenic activities is an increasing problem worldwide. In this study, the contamination and ecological risk by several trace elements were evaluated along a 100-km stretch of the San Jorge River in Colombia, impacted by different mining activities. The increase of average concentration levels and range of trace elements in sediments (in μg/g) was as follows: Cu 6656 (454–69,702) > Cd 1159 (0.061–16,227) > Zn 1064 (102–13,483) > Ni 105 (31–686) > Pb 7.2 (5.1–11.7) > As 1.8 (1.0–3.2) > Hg 0.31 (0.12–1.37). Results showed that surface sediments could be classified as very high ecological risk index (RI > 600), associated with high contamination of Hg, Cd, and Cu, in stations close mining activities. Values for pollution load index indicate an environmental deterioration (PLI > 1), and sediment quality guidelines (SQGs) suggested that Cu, Ni, Zn, and Hg caused adverse biological effects. We further used pollution indices such as contamination factor (CF), enrichment factor (EF), and geoaccumulation index (Igeo) to assess the extent of contamination. According to these indices, discharges of hazardous chemicals over many years have resulted in a high degree of pollution for Cu, Pb, and Cd, with critical values in stations receiving wastes from mining activities. Multivariate statistical analysis suggested that Hg, Cd, Cu, and Zn derived from gold and coal mining, Ni and As were related from the mining of ferronickel and coal, respectively, whereas the high Pb load was attributed to diffuse source of pollution. In sum, our study provided the first detailed database on metal concentration and ecological risks to organisms in sediments of the San Jorge River Basin, and the current results also suggested future research for public health action.

     

Inconsistency and comprehensiveness of risk assessments for heavy metals in urban surface sediments

Numerous indices have been developed to assess environmental risk of heavy metals in surface sediments, including the total content based geoaccumulation index (I_geo), exchangeable fraction based risk assessment code (RAC), and biological toxicity test based sediment quality guidelines (SQGs). In this study, the three indices were applied to freshwater surface sediments from 10 sections along an urbanization gradient of the Grand Canal, China to assess the environmental risks of heavy metals (Cu, Pb, Zn, Cd, and Cr) and to understand discrepancies of risk assessment indices and urbanization effects regarding heavy metal contamination. Results showed that Cd, Zn, and Pb were the most enriched metals in urban sections assessed by I_geo and over 95% of the samples exceeded the Zn and Pb thresholds of the effect range low (ERL) of SQGs. According to RAC, Cu, Zn, Cd, and Cr had high risks of adversely affecting the water quality of the Grand Canal due to their remarkable portions of exchangeable fraction in surface sediment. However, Pb showed a relative low risk, and was largely bounded to Fe/Mn oxides in the urban surface sediments. Obviously, the three assessment indices were not consistent with each other in terms of predicting environmental risks attributed to heavy metals in the freshwater surface sediments of this study. It is recommended that risk assessment by SQGs should be revised according to availability and site specificity. However, the combination of the three indices gave us a comprehensive understanding of heavy metal risks in the urban surface sediments of the Grand Canal.     

Metal accumulation in the greentail prawn, Metapenaeus bennettae, in Sydney and Port Hacking estuaries, Australia

Metal concentrations of the inshore greentail prawn, Metapenaeus bennettae, and surface sediments from locations within Sydney estuary and Port Hacking (Australia) were assessed for bioaccumulation and contamination. The current study aimed to assess metal concentrations in prawn tissue (tail muscle, exoskeleton, hepatopancreas and gills), relate whole body prawn tissue metal concentrations to sediment metal concentrations and animal size, as well as assess prawn consumption as a risk to human health. Metal concentrations were highest in sediment and prawns from contaminated locations (Iron Cove, Hen and Chicken Bay and Lane Cove) in Sydney estuary compared with the reference estuary (Port Hacking). Concentrations in sediments varied considerably between sites and between metals (As, Cd, Cr, Cu, Ni, Pb and Zn), and although concentrations exceeded Interim Sediment Quality Guideline-Low values, metals (As, Cd, Cr, Cu, Ni, Pb and Zn) were below Australian National Health and Medical Research Council human consumption guidelines in prawn tail muscle tissue. Metal concentrations in prawn tail muscle tissue were significantly different (p?≤?0.05) amongst locations for Pb, Zn and Cd, and metal concentrations were generally highest in gills tissue, followed by the hepatopancreas, exoskeleton and tail muscle. The exoskeleton contained the highest Sr concentration; the hepatopancreas contained the highest As, Cu and Mo concentrations; and the gills contained the highest Al, Cr, Fe and Pb concentrations. Concentrations of Pb, As and Sr were significantly different (p?≤?0.05) between size groups amongst locations.     

西湖沉积物中元素分布特征研究

分析测定了西湖沉积物（２０ｍ钻探样品）不同层次中Ｆｅ、Ａｌ、Ｃａ、Ｍｇ、Ｔｉ、Ｍｎ、Ｃｕ、Ｚｎ、Ｃｄ、Ｎｉ、Ｈｇ、Ｃｒ、Ａｓ、Ｃｏ、Ｖ等１５种元素的含量，并根据元素的分布特征探讨了西湖沉积物的形成发展及重金属污染状况。     

Selected trace metals in oysters (Crassostrea iridescens) and sediments from the discharge zone of the submarine sewage outfall in Mazatlán Bay (southeast Gulf of California): chemical fractions and bioaccumulation factors

Concentrations of Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn in the soft tissue of Crassostrea iridescens and the associated surface sediments (bulk and bioavailable metal concentrations) from an area influenced by a sewage outfall in Mazatlán Bay (southeast Gulf of California), were determined by atomic absorption spectrophotometry. Significant spatial differences in metal concentrations in both the bulk and bioavailable forms in the sediments were identified. An enrichment of Cu, Ni, Pb and Zn in sites located on a south-north transect was detected indicating a dominant influence of the sewage outfall toward the north. C. iridescens accumulated more Zn, Cu, Ni, Fe, Cd; and less Mn, Cr and Pb than were bioavailable in the sediments, as measured using conventional extraction analysis. The degree of enrichment and the bioavailable metal concentrations in the sediments of the south portion of Mazatlán Bay is discussed. The potential ability of C. iridescens as a biomonitor of metallic pollutants is postulated.     

Comprehensive assessment of toxic metals in urban and suburban street deposited sediments (SDSs) in the biggest metropolitan area of China

A set of toxic metals, i.e. As, Hg, Pb, Cd, Cu, Zn, Ni and Cr, in urban and suburban SDSs were investigated comparatively in the biggest metropolitan area of China, Shanghai. Results showed that all of the metals except As were accumulated greatly, much higher than background values. Geo-accumulation index indicated that metal contamination in urban SDSs was generally heavier than that in suburban SDSs. Potential ecological risk index demonstrated that overall risks caused by metals were considerable. Cd contributed 52% to the overall risk. Multivariate statistical analysis revealed that in urban SDSs, Zn, Ni, Cd, Pb, Cu and Cr were related to traffic and industry; coal combustion led to elevated levels of Hg; soil parent materials controlled As contents. In suburban SDSs, Pb, Cu, As and Cd largely originated from traffic pollution; Zn, Ni and Cr were associated with industrial contaminants; Hg was mainly from domestic solid waste.     

Evaluation of the anthropogenic influx of metal and metalloid contaminants into the Moulay Bousselham lagoon, Morocco, using chemometric methods coupled to geographical information systems

Superficial and cored sediment samples from the Moulay Bousselham lagoon and sub-watershed were analyzed for Al, Fe, Cu, Zn, Pb, Mn, Ni, Cr, As, Hg, and Cd. The temporal and spatial distributions of the main contamination sources of heavy metals were identified and described using chemometric and geographic information system (GIS) methods. Sediments from coastal lagoons near urban and agricultural areas are commonly contaminated with heavy metals, and the concentrations found in surface sediments are significantly higher than those from 50–100 years ago. The concentrations of these elements decrease sharply with depth in the sediment column, and the elements are preferentially enriched in the <2-μm-sized fraction of the sediment. The zones of enhanced risk of heavy metals were detected by means of GIS-based geostatistical modeling. According to sediment pollution indices and statistical analysis, heavy metals (Pb, Cu, Ni, Zn, Cr, and Hg) that pose a risk have become largely enriched in the lagoon sediments during the recent period of agricultural intensification.     

Processes controlling metal transport and retention as metal-contaminated groundwaters efflux through estuarine sediments

Factors affecting the transport and retention of Cd, Cr, Cu, Ni, Pb and Zn in acidic groundwaters as they pass through estuarine sediments were investigated using column experiments. Acidic groundwaters caused the rapid dissolution of iron sulfide (AVS) and other iron and manganese phases from sediments that are important for metal binding and buffering. Metal breakthrough to overlying water occurred in the order of Ni>Zn>Cd>Cu>Cr/Pb. Metal transport increased as the sediment permeability increased, reflecting the low resistance to flow caused by larger sand-sized particles and the decreased abundance of metal adsorption sites on these materials. Metal mobility increased as the groundwater pH decreased, as flow rate or metal concentrations increased, and as the exposure duration increased. Groundwater Cr and Pb were promptly attenuated by the sediments, the mobility of Cu was low and decreased rapidly as sediment pH increased above 4.5, while Cd, Ni and Zn were the most easily transported to the surface sediments and released to the overlying waters. For groundwaters of pH 3, metal migration velocities through sandy sediments were generally 0.5-2% (Cr, Pb), 1-6% (Cu) and 4-13% (Cd, Ni, Zn) of the total groundwater velocity (9-700 m/yr). The oxidative precipitation of Fe(II) and Mn(II) in the groundwaters did not affect metal mobility through the sediments. The results indicated that the efflux of acidic and metal-contaminated groundwater through estuarine sediments would affect organisms resident in sandy sediments more greatly than organisms resident in fine-grained, silty, sediments.     

Mobility of heavy metals from polluted sediments of a semi-enclosed basin: in situ benthic chamber experiments in Taranto’s Mar Piccolo (Ionian Sea,Southern Italy)

In situ benthic flux experiments were conducted at two stations in the Mar Piccolo of Taranto (Italy), one of the most industrialised and contaminated coastal areas of the Mediterranean. Sediments of the two stations are notably different in their trace metal content, with a station closer to a Navy harbour showing higher mean concentrations of almost all investigated metals (Al, As, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, V and Zn). Conversely, both stations are characterised by significant Hg contamination, compared to the local baseline. Results of a sequential extraction scheme on surface sediments suggest a relatively scarce mobility of the examined metals (Zn > Ni > Cr > As > Cu > Pb). A Hg-specific extraction procedure showed that most of the element (93.1 %) occurs in a fraction comprising Hg bound to Fe/Mn oxi-hydroxides. Reduction of these oxides may affect Hg remobilisation and redistribution. Porewater profiles of dissolved trace metals were quite similar in the two sites, although significant differences could be observed for Al, Cu, Fe and Hg. The highest diffusive fluxes were observed for As, Fe and Mn. Mobility rates of several trace elements (Al, As, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, V and Zn) were directly measured at the sediment–water interface. Results from benthic in situ incubation experiments showed increasing dissolved metal concentrations with time, resulting in higher fluxes for Cu, Fe, Hg, V and Zn in the most contaminated site. Conversely, fluxes of Mn, Ni and Pb were comparable between the two stations. The estimated flux of Hg (97 μg m^?2 day^?1) was the highest observed among similar experiments conducted in other highly contaminated Mediterranean coastal environments. Benthic fluxes could be partially explained by considering rates of organic matter remineralisation, dissolution of Fe/Mn oxy-hydroxides and metal speciation in sediments. Seasonal and spatial variation of biogeochemical parameters can influence metal remobilisation in the Mar Piccolo area. In particular, metals could be promptly remobilised as a consequence of oxygen depletion, posing a serious concern for the widespread fishing and mussel farming activities in the area.     

Spatial distribution and ecological risk assessment of sediment metals in a highly industrialized coastal zone southwestern Taiwan

Spatial variations of Cr, Cu, Hg, Ni, Pb, and Zn in the surface sediments from 34 stations of the Kaohsiung coastal zone southwestern Taiwan were studied to address the current pollution status, sediment quality, and potential ecological risk. The study revealed that the concentrations of sediment metals in Kaohsiung Harbor were alarmingly high compared to the other region of Kaohsiung coast. The concentrations of Cr, Cu, Hg, Ni, Pb, and Zn in the harbor sediments were as high as 351, 247, 1.93, 61.8, 60.9, and 940 mg kg⁻¹, respectively. The current situation of metal pollution was assessed by different pollution indices and results showed moderate to severe enrichment of Cu, Hg, and Zn in the harbor sediments. According to the degree of contamination, pollution load index, and contamination severity index, the sediments from the inner Kaohsiung Harbor show high degree and high severity of metal contamination, while the rest of Kaohsiung coastal areas show uncontaminated or low-level pollution. Results of mean ERM quotient and potential ecological risk index also indicated that the harbor sediments posed a 49% probability of biological toxicity and very high ecological risk. The toxic units indicated that the negative biological effects of the six metals in the harbor sediments were Zn > Cu > Cr > Ni > Hg > Pb. In contrast to Kaohsiung Harbor as a trap where considerable amount of anthropogenic metal loadings accumulated in sediments, low metal concentrations were observed in most Kaohsiung coastal sediments. It probably resulted from the limited fine-grained sediment deposition. In the wave-dominated Kaohsiung coastal zone, fine-grained sediments associated with polluted metals tend to be easily resuspended and transported offshore via waves and wave-induced currents. The results of this study can provide valuable information for river and coastal zone management.

     

Heavy metal pollution in sediments of the Pasvik River drainage

The purpose of this paper is to study the regional impacts of heavy metals (Ni, Cu, Co, Zn, Cd, Pb, Hg) on the watershed of the Pasvik River. On the basis of sediment investigations at 27 stations of the watershed, background concentrations of the heavy metals, vertical distribution of heavy metals in sediments, heavy metal concentrations in surface sediments, contamination degree, and risk index were determined. The atmospheric emissions of Ni, Cu, Co, Zn, Cd and Hg from the smelters and waste waters from tailing dams and mines of the Pechenganickel Company are likely to be the main sources of increasing concentrations observed in recent sediments of the lower river reaches. Lead showed a different pattern from the other heavy metals--increasing Pb concentrations in the upper sediment layers towards the Norwegian side.     

Survey of heavy metal pollution and assessment of agricultural soil in Yangzhong district, Jiangsu Province, China

We investigated concentrations of Hg, Cd, Pb, Zn, Cu, As, Ni, and Cr in samples of soil, cereal, and vegetables from Yangzhong district, China. Compared to subsoils, the sampled topsoils are enriched in Hg, Cd, Cu, Pb, Zn, and As. High levels of Cd and Hg are observed in most agricultural soils. Concentrations of Cr and Ni show little spatial variation, and high Cu, Pb, and Zn contents correspond well to areas of urban development. High As contents are primarily recorded at the two ends of the sampled alluvion. The contents of Cd, Hg, and total organic carbon (TOC) increase gradually to maximum values in the upper parts of soil profiles, while Cr and Ni occur in low concentrations within sampled profiles. As, Pb, Cu, and Zn show patterns of slight enrichment within the surface layer. Compared to data obtained in 1990, Cd and Hg show increased concentrations in 2005; this is attributed to the long-term use of agrochemicals. Cr and Ni contents remained steady over this interval because they are derived from the weathering of parent material and subsequent pedogenesis. The measured As, Cu, Pb, and Zn contents show slight increases over time due to atmospheric deposition of material sourced from urban anthropogenic activity. Low concentrations of heavy metals are recorded in vegetables and cereals because the subalkaline environment of the soil limits their mobility. Although the heavy metal concentrations measured in this study do not pose a serious health risk, they do affect the quality of agricultural products.     

Dating of marine sediments and time evolution of heavy metal concentrations in the Bay of Cádiz, Spain

In this paper the time evolution of heavy metal concentration of Pb, Zn, Cd and Hg, in the sediments of the Bay of Cádiz (southwest of Spain) is studied during the past century, as a result of the industrial influence in the zone. The study has been performed using sedimentary profiles that have been extracted from the seabed. The measurement of 210Pb and 137Cs radionuclides has provided the dating of the sediment layers, up to a depth corresponding to the age of 115 years. The relative sedimentation rates obtained are around 0.2 cm/year. The 137Cs activity profile reflects the concentration of this radionuclide in the atmosphere and into aquatic systems during the second half of the twentieth century. This profile has been used to ratify the results provided by the 210Pb dating method.     

Distribution,Sources, and Possible Adverse Biological Effects of Trace Metals in Surface Sediments of a Tropical Estuary

Metal (Cu, Zn, Pb, Cd, Ni, Co, and Fe) contamination in sediments from a tropical estuary (Ébrié Lagoon, Ivory Coast) was assessed using pollution indices, multivariate analyses and sediment quality guidelines (SQGs). The results demonstrate that increased input of the studied metals occurred over the past 6 years compared to that from 20 years ago, due to rapid population growth, along with the increase of industrial and agricultural activities in the vicinity of the estuary. Ébrié Lagoon was also found to be one of the most contaminated tropical coastal estuaries. Very high average total organic carbon (TOC) content was found (1.9–3.70%) with significant spatial variation as a result of the influence of anthropogenic activities. This study also found that TOC plays an important role in the distribution of Cu, Zn, Co, and Cd in the Ébrié Lagoon sediments. Moderate to high sediment contamination was observed for Cd and Cu, moderate contamination was observed for Zn and Pb, while low contamination was observed for Ni, Co, and Fe. Cluster analysis (CA) and principal component analysis (PCA) investigation revealed that Cu, Zn, Cd, and Co result mainly from anthropogenic sources while Pb, Ni, and Fe may be of natural origin. The pollution-loading index (PLI) indicated that all of the sites close to wastewater discharges were highly polluted. The sediments are likely to be an occasional threat to aquatic organisms due to Cu, Zn, Pb, Cd, and Ni contents, based on the SQGs approach.     

Using Various Guidelines and Approaches for the Assessment of Marine Sediment Quality

The aim of the present study is to explore the implications of using different quality guidelines and approaches for marine sediment characterization. As a case study, the sediments of Souda Port, Crete, Greece, were examined in order to assess heavy metal contamination and to estimate anthropogenic contribution and potential adverse effects to aquatic environment, as well as to indicate feasible future management options. The concentration of six heavy metals (Co, Cr, Cu, Ni, Pb, and Zn) was determined and compared with different sediment quality guidelines (SQGs) and regional/national criteria established for dredged material characterization and management. In addition, the geo-accumulation index (I_geo) was calculated and the sediments toxicity and heavy metals bioavailability were determined. Based on the experimental data, it is deduced that Souda sediments are characterized as moderately polluted only in terms of Co, Ni, and Pb.