Effect of Salinity on the Tolerance to Toxic Metals and Oxyanions in Native Moderately Halophilic Spore-forming Bacilli

Summary Ten moderately halophilic spore-forming bacilli were isolated from saline soils in Iran and their intrinsic high-level resistance to chromate, arsenate, tellurite, selenite, selenate and biselenite was identified by an agar dilution method. Minimum inhibitory concentration (MIC) for each oxyanion was determined. All isolates were resistant to higher concentrations of arsenate. The resistance level of the isolates to selenooxyanions was between 10 and 40 mM. Maximum and minimum tolerance against oxyanions was seen in selenite and biselenite, respectively. Although toxic metal resistance in the isolates was not different from non-halophilic bacteria that has been reported, unusual resistance to arsenate (250 mM), sodium chromate (75 mM) and potassium chromate (70 mM) was observed. The results obtained in this study revealed that all isolates were obviously susceptible to silver, nickel, zinc and cobalt, while seven isolates were resistant to lead. Susceptibility to copper and cadmium varied among the isolates. Silver had the maximum toxicity, whereas lead and copper showed minimum toxicity. The impact of salinity on the toxicity of oxyanions was also studied. Our results showed that in general an increase in salinity from 5% (w/v) to 15% (w/v) enhanced tolerance to toxic oxyanions.     

Detection of heavy metal ion resistance genes in Gram-positive and Gram-negative bacteria isolated from a lead-contaminated site

Resistance to a range of heavy metal ions wasdetermined for lead-resistant and other bacteria whichhad been isolated from a battery-manufacturing sitecontaminated with high concentrations of lead. Several Gram-positive (belonging to the genera Arthrobacter and Corynebacterium) andGram-negative (Alcaligenes species) isolateswere resistant to lead, mercury, cadmium, cobalt,zinc and copper, although the levels of resistance tothe different metal ions were specific for eachisolate. Polymerase chain reaction, DNA-DNAhybridization and DNA sequencing were used to explorethe nature of genetic systems responsible for themetal resistance in eight of the isolates. SpecificDNA sequences could be amplified from the genomic DNAof all the isolates using primers for sections of themer (mercury resistance determinant on thetransposon Tn501) and pco (copperresistance determinant on the plasmid pRJ1004) geneticsystems. Positive hybridizations with mer andpco probes indicated that the amplified segmentswere highly homologous to these genes. Some of thePCR products were cloned and partially sequenced, andthe regions sequenced were highly homologous to theappropriate regions of the mer and pcodeterminants. These results demonstrate the widedistribution of mercury and copper resistance genes inboth Gram-positive and Gram-negative isolates obtainedfrom this lead-contaminated soil. In contrast, theczc (cobalt, zinc and cadmium resistance) andchr (chromate resistance) genes could not beamplified from DNAs of some isolates, indicating thelimited contribution, if any, of these genetic systemsto the metal ion resistance of these isolates.     

Survey of metal tolerance in moderately halophilic eubacteria.

The tolerance patterns, expressed as MICs, for 250 moderately halophilic eubacteria to 10 heavy metals were surveyed by using an agar dilution method. The moderate halophiles tested included 12 culture collection strains and fresh isolates representative of Deleya halophila (37 strains), Acinetobacter sp. (24 strains), Flavobacterium sp. (28 strains), and 149 moderately halophilic gram-positive cocci included in the genera Marinococcus, Sporosarcina, Micrococcus, and Staphylococcus. On the basis of the MICs, the collection strains showed, overall, similar responses to silver, cobalt, mercury, nickel, lead, and zinc. All were sensitive to silver, mercury, and zinc and tolerant of lead. The response to arsenate, cadmium, chromium, and copper was very heterogeneous. The metal susceptibility levels of the 238 freshly isolated strains were, in general, very heterogeneous among the four taxonomic groups as well as within the strains included in each group. The highest toxicities were found with mercury, silver, and zinc, while arsenate showed the lowest activity. All these strains were tolerant of nickel, lead, and chromium and sensitive to silver and mercury. Acinetobacter sp. strains were the most heavy-metal tolerant, with the majority of them showing tolerance of eight different metal ions. In contrast, Flavobacterium sp. strains were the most metal sensitive. The influence of salinity and yeast extract concentrations of the culture medium on the toxicity of the heavy metals tested for some representative strains was also studied. Lowering the salinity, in general, led to enhanced sensitivity to cadmium and, in some cases, to cobalt and copper. However, increasing the salinity resulted in only a slight decrease in the cadmium, copper, and nickel toxicities.(ABSTRACT TRUNCATED AT 250 WORDS)     

Survey of metal tolerance in moderately halophilic eubacteria

The tolerance patterns, expressed as MICs, for 250 moderately halophilic eubacteria to 10 heavy metals were surveyed by using an agar dilution method. The moderate halophiles tested included 12 culture collection strains and fresh isolates representative of Deleya halophila (37 strains), Acinetobacter sp. (24 strains), Flavobacterium sp. (28 strains), and 149 moderately halophilic gram-positive cocci included in the genera Marinococcus, Sporosarcina, Micrococcus, and Staphylococcus. On the basis of the MICs, the collection strains showed, overall, similar responses to silver, cobalt, mercury, nickel, lead, and zinc. All were sensitive to silver, mercury, and zinc and tolerant of lead. The response to arsenate, cadmium, chromium, and copper was very heterogeneous. The metal susceptibility levels of the 238 freshly isolated strains were, in general, very heterogeneous among the four taxonomic groups as well as within the strains included in each group. The highest toxicities were found with mercury, silver, and zinc, while arsenate showed the lowest activity. All these strains were tolerant of nickel, lead, and chromium and sensitive to silver and mercury. Acinetobacter sp. strains were the most heavy-metal tolerant, with the majority of them showing tolerance of eight different metal ions. In contrast, Flavobacterium sp. strains were the most metal sensitive. The influence of salinity and yeast extract concentrations of the culture medium on the toxicity of the heavy metals tested for some representative strains was also studied. Lowering the salinity, in general, led to enhanced sensitivity to cadmium and, in some cases, to cobalt and copper. However, increasing the salinity resulted in only a slight decrease in the cadmium, copper, and nickel toxicities.(ABSTRACT TRUNCATED AT 250 WORDS)     

Reduction of marine phytoplankton reproduction rates by copper and cadmium

The reproduction rates of 38 clones of marine phytoplankton were measured in media in which free cupric ion activity was controlled at different levels using a NTA-cupric ion buffer system. The major trend among species in their resistance to copper toxicity was a phylogenetic one, with cyanobacteria being the most sensitive, diatoms the least sensitive, and coccolithophores and dinoflagellates intermediate in sensitivity. The reproduction rates of most of the cyanobacteria were reduced at cupric ion activities above 10⁻¹² M, while most eukaryotic algae still had maximum reproduction rates at 10⁻¹¹ M. Four species, Emiliana huxleyi (Lohm.) Hay & Mohler, Skeletonema costatum (Grev.) Cleve, Thalassiosira pseudonana (Hustedt) Hasle & Heimdal and Thalassiosira oceanica (Hustedt) Hasle were particularly resistant to copper, being able to reproduce well at the highest cupric ion activities tested, 10^−9.5 M and 10^−9.2 M. There was no major difference, however, between neritic and oceanic species in their sensitivity to copper.The sensitivity of 20 species of marine phytoplankton to free cadmium ion activity was measured in a similar manner using an NTA-cadmium ion buffer system. As observed with copper, the prokaryotic cyanobacteria were the most sensitive to cadmium toxicity, diatoms were the least sensitive, and coccolithophores and dinoflagellates were intermediate. All cyanobacteria tested were dead at a cadmium ion activity of 10^−9.3 M whereas the reproduction rates of most of the eukaryotic algae were not reduced significantly until 10^−8.3 M.Comparison of these data with natural concentrations in sea water implies that cadmium is not an important ecological factor in unpolluted waters but natural copper concentrations may inhibit the reproduction of some phytoplankton species, especially cyanobacteria, in upwelled sea water. Copper may influence the seasonal succession of species.     

Levels of zinc, cadmium and lead in some marine algae from Aqaba-Red Sea

Jordan has witnessed a rapid industrial development in the last twenty years. This has lead to the release of waste materials or pollutants into the marine environment, particularly nearby Aqaba Port. The present study investigates the levels of zinc, cadmium and lead in four brown algae, three red algae and four green algal species collected from Aqaba. Three different levels of lead and zinc concentrations were found: the highest level of both metals is exhibited among brown algae; intermediate level is exhibited among red algae and the lowest level is seen among the green algae. Very low concentrations of cadmium were found in all examined algal species. The results indicate that the brown algal species Cystosira myrica, Sargassum asperifolium, Sargassum neglectum, and Sargassum subrepandum always contain the highest concentrations of lead and zinc, but these algae are less contaminated than brown algae from industrial European seas.     

Species associations and metal contents of algae from rivers polluted by heavy metals

SUMMARY. The algal flora of the Rivers Hayle and Gannel. whieh drain copper and lead mining regions of Cornwall, are described and compared. Although fluctuations in the rarer members of the algal communities were observed, the dominant filamentous algae at most sites did not change over the year of study. Both the total algal abundance and the number of species were depressed at high metal sites. Associations of species that were evident in field samples were confirmed and correlated with water metal levels by principal component analysis. The close similarity between the flora of similar sites on the copper-polluted River Hayle and the lead-polluted River Gannei implies that the degree of metal pollution, rather than the polluting metal per se, determines the species present. All mine sites were characterized by a Microspora -community whereas a Zygnemales community of Spirogyra and Mougeotia species was typical of low metal pollution. Moderately polluted sites downsteam of the mines had an intermediate flora of Zygnemaies, Microsporales. Ulotrichales and gelatinous Volvocales and Tetrasporales species. No species could be said to invariably indicate metal pollution; the most abundant species at highly contaminated sites were also those with the widest distributions. Field samples of filamentous algae (mainly Spirogyra, Zygogonium, Mougeotia and Microspora species) contained metal concentrations several orders of magnitude greater than ambient levels. For copper and iron, both algal metal contents and concentration ratios (μg g-1 algae/μg ml-1 water) were positive functions of water metal levels, although algal iron appeared to plateau at water concentrations of about 1 mg ml-1. Algal lead concentrations, but not the concentration ratios, also were positively correlated with water lead levels. Thus, for these three metals, the algal metal contents were indicative of ambient conditions. In contrast, algal zinc concentrations were nearly constant and consequently the concentration ratios for this metal were inversely related to water levels. This result suggests that unlike the uptake of other metals, uptake of zinc by these aigae may be strictly regulated.     

Application of in situ cage cultures of phytoplankton for monitoring heavy metal pollution in two Norwegian fjords

A simple apparatus for in situ use of the cage culture technique for growing planktonic algae has been developed and used to follow growth and metal uptake of three diatoms in two Norwegian fjords polluted by heavy metals. Continuous pumping of sea water through a chelating resin and continuous water sampling was used to obtain average values for dissolved and particulate heavy metal content at the various test sites. The metals investigated (zinc, copper, lead, cadmium, and mercury) showed differences in the proportion of dissolved to particulate fractions.The three species of diatoms tested gave systematic growth responses to heavy metal pollution in the moderately polluted fjord; one alga died, one showed reduced growth rate at the more contaminated site compared with those at less polluted sites, while the most tolerant alga was apparently not affected. In the heavily polluted fjord only the most tolerant alga survived, showing decreasing growth rate with increasing pollution.Uptake of heavy metals increased generally with increasing heavy metal content in the sea water. The contents in the algae grown in the most polluted fjord were much higher than those obtained in the less polluted fjord, which were higher than the contents reported for algae from non-contaminated areas.     

The effect of industrial pollution on copper,lead, zinc and cadmium concentration in xylem rings of resistant (Populus marilandica) and sensitive (P. balsamifera) species of poplar

Summary The patterns of radial distribution of copper, lead, zinc and cadmium in two species of poplar, growing under polluted conditions near a copper smelter, were investigated. Populus marilandica is very resistant to such pollution, but P. balsamifera is sensitive. Soils were examined for concentrations of these metals, and only a surface layer of soil was polluted with copper and lead. P. balsamifera absorbed much more cadmium than P. marilandica under the same conditions. P. marilandica shows a pattern of copper radial distribution with significantly increased concentration of the meral in the layers in the middle of the trunk compared to P. balsamifera. Zinc and cadmium have a pattern with increased metal concentration from the bark towards the pith of the tree. In the case of copper and lead the pattern indicates increased concentrations of these metals in the rings adjacent to the pith while the pattern in other parts of the tree remains unconfirmed due to poor precision. Poplar xylem seems to accumulate zinc and cadmium while the content of lead in xylem is much lower than in the adjacent soil.This paper is dedicated to the memory of Professor Stefan Bialobok, Institute of Dendrology, Polish Academy of Sciences in Kornik     

Lead and copper effects on lipid metabolism in cultured lichen photobionts with different phosphorus status

We evaluated the ability of heavy metals (copper, lead) to alter lipid metabolism in four algal lichen photobionts following short term exposure. Metal concentrations (10 microM) were equivalent to environmentally relevant levels that have been reported to have effects on intact algae. The algae were grown under normal or deficient phosphate conditions to assess any interactions with the heavy metal stress. Given the frequent sensitivity of lichens to copper and lead, there were surprisingly small changes on lipid metabolism, as assessed by radiolabelling from [1-14C]acetate. The main effects, which were seen in a number of cases, were an overall inhibition of total lipid labelling and a relative increase in the labelling of triacylglycerols in the non-polar fraction. Both of these changes can be viewed as reflecting general toxicity of heavy metals. The Coccomyxa photobiont species were more sensitive than Trebouxia species, which fits with the general distribution of the latter in lichens inhabiting harsh environments.     

A morphometric analysis of algal response to low dose,short-term heavy metal exposure

Summary Three algae,Melosira granulata, Fragilaria capucina, andAnacystis cyanea, collected as part of a natural phytoplankton assemblage were found to differ in their cytological responses to low dose short-term exposure to copper and lead. In general, all were more sensitive to copper than to lead.Fragilaria was more sensitive to both metals than the other species examined. Most immediate changes in relative volume categories can be ascribed to changes in vacuole volume that are most likely the result of changes in membrane permeability. There was some degree of accommodation in all three species at 24 hours. These results are discussed in view of the natural environment of the algae, as well as in relationship to previous studies.     

Certain metals in three coastal algae from Ras Beirut waters

In July and August of 1977, samples of Halimeda tuna (Ellis & Solander) Lamouroux, Calpomenia sinuosa (Mertens & Roth) Derbes & Solier, and Pterocladia pinnata (Hudson) Papenfuss were collected along the coast of Ras Beirut, Lebanon and analyzed for lead, cadmium, copper, nickel and iron. The metal concentrations were quite similar in all three algae, except for iron, which was markedly higher in C. sinuosa, and lead, which was considerably lower in P. pinnata than in the other two species. These metal levels, particularly of lead, copper, and nickel seem to be rather high and may, in part, be due to untreated domestic sewage, industrial effluent, automobile exhaust, and outdoor incineration.Because little is known about the uptake, retention, use and tolerance of heavy metals by marine algae, more work in this field of study is needed.     

Trace metals with seasonal considerations in coastal algae and molluscs from Beirut,Lebanon

Twelve species of intertidal algae and molluscs from Ras Beirut, Lebanon have been investigated for their heavy metal content. Atomic absorption spectrophotometric analysis showed that the algae and molluscs concentrated similar levels of most metals. However, of the twelve organisms, Brachydontes variabilis had the highest copper, Patella coerulea and Colpomenia sinuosa had the highest iron, and Pinctada radiata had the highest zinc values with rather elevated cadmium. Cystoseira spinosa concentrated the lowest levels of zinc, and nickel was variable in all organisms.Seasonal comparisons of metal levels were also considered in three algae and three molluscs studied previously. The only signs of seasonal variation in the algae were with nickel and iron in Halimeda tuna and lead in Pterocladia pinnata. Both cadmium and lead were generally lowest in the spring samples of the algae. Lead and nickel concentrations in the molluscs were generally highest in the summer and lowest in the spring. Slight seasonal trends were seen with lead and iron in Brachydontes variabilis and lead in Monodonta turbinata.The lack of comparison data from the Mediterranean, particularly the eastern basin, makes it essential that studies of this nature be continued.     

Molasses melanoidin-like products enhance phytoextraction of lead through three Brassica species

Previously, it has been suggested that melanoidin-like products (MLP) from sugarcane molasses may accelerate copper phytoextraction. In this study, we evaluated the facilitatory effect of MLP on phytoextraction in a medium including cadmium or lead, the concentrations of which were adjusted around the regulation values of the Soil Contamination Countermeasures Act in Japan. Three Brassica species were tested based on their fast growth, high biomass productivity, and high heavy metal absorption. Both biomass and lead uptake in the nutrient medium with 1 mM lead nitrate were significantly increased by the addition of MLP, and almost all of the lead was accumulated in the root tissue. Therefore, MLP were able both to detoxify lead ions and to improve their bioavailability in Brassica species. In contrast, only these species with MLP or citric acid survived in the nutrient medium with 1 mM cadmium sulfate. The phytoextraction of cadmium using these species was therefore impractical under the Act.     

Changes in Endogenous Fatty Acids and Lipid Synthesis Associated with Copper Pollution in Fucus spp

Fatty acid synthesis from [¹⁴C]acetate was studied in the brownalgae, Fucus serratus and Fucus vesiculosus, which had beencollected from sites polluted by run-off from old copper mines.Algae collected from areas exposed to copper in situ showedchanges in the pattern of fatty acids labelled in vitro withan increase in oleate labelling and a decrease in palmitatelabelling in both species. The endogenous fatty acid patternsfor algae from polluted sites also differed from those fromunpolluted sites. Algae from sites polluted by copper containedmore oleate than samples from non-polluted sites. In addition,there were decreases in the polyunsaturated fatty acids -linolenate,octadecatetraenoate and docosapentaenoate for copper-exposedalgae. These differences are discussed with regard to the possiblemetabolic site of copper's action and to the relative toleranceof Fucus spp. to high environmental dissolved copper levels. Key words: Fucus spp., Copper pollution, Fatty acid synthesis     

The effect of industrial pollution on Zinc,Cadmium and Copper concentration in the xylem rings of Scot's pine (Pinus sylvestris L.) and in the soil

Summary The concentrations of heavy metals, including zinc, cadmium, and copper were studied in the xylem rings of Pinus sylvestris trees and in the soil of three heavily polluted sites and one non-polluted site. There was a high correlation between the concentration of heavy metals in the soil and recently produced xylem rings. However, the initiation of industrial pollution did not abruptly increase heavy metal concentrations in the xylem rings. There was a high correlation between the zinc content of trees growing near a zinc smelter and the values obtained by other investigators for pine trees growing on the same site.     

乐安河—鄱阳湖湿地植物群落特征及其优势植物对重金属Cu、Pb、Cd的富集

选择乐安河—鄱阳湖湿地典型植物群落,采用重要值方法评价各样点植物群落特征并筛选出典型优势植物,通过室内理化测试分析不同生境中优势植物植株及其根区土壤中重金属Cu、Pb、Cd的含量;采用生物富集系数(BCF)方法评价不同优势植物对重金属Cu、Pb、Cd的富集特性。结果表明:研究区湿地植物以草本为主,在各样点共发现124种物种,包括蕨类植物2科2属2种,种子植物40科97属122种,并从中筛选出羊蹄、红蓼、鼠曲草、紫云英、苎麻等5种富集能力较强的优势植物;植物根区土壤中的Cu、Cd含量均超过土壤环境质量三级标准,而且Cu、Cd的最高含量分别为824.03、5.03 mg·kg-1;不同优势植物对Cu、Pb、Cd等3种重金属元素中的1种或2种表现出较强的富集能力,其中优势物种红蓼对Cu具有较强的富集能力,含Cu量最高为148.80 mg·kg-1,另一种优势物种鼠曲草对三种元素的生物富集系数均较高,且对Cd的最高富集含量为15.17 mg·kg-1,对Cd的生物富集系数最高值为19.14,高于其他植物10倍以上,鼠曲草对重金属Cd具有富集植物的基本特征,且对Cu和Cd具有共富集特征并具有较高的耐性,紫云英、羊蹄等对Cd的富集能力也较强。上述5种优势植物种群对鄱阳湖湿地Cu、Pb、Cd等重金属污染物的生态修复具有一定参考价值,可作为鄱阳湖湿地重金属污染修复植物的选择对象。     

Copper accumulation, synthesis of ascorbate and activation of ascorbate peroxidase in Enteromorpha compressa (L.) Grev. (Chlorophyta) from heavy metal-enriched environments in northern Chile

Enteromorpha compressa is the dominant species in coastal areas of northern Chile receiving copper mine wastes. Copper remains as the main heavy metal in these coastal waters and it is accumulated in E. compressa growing at the impacted sites. Algae from these sites showed higher levels of lipoperoxides than from non‐impacted sites, which suggests the occurrence of cellular damage resulting from oxidative stress. The strong activation of ascorbate peroxidase detected in this study probably occurs in order to buffer this oxidative stress. Unexpectedly, the activity of glutathione reductase, normally coupled to ascorbate peroxidase activity, was not affected by the chronic exposure to the mine wastes. Moreover, catalase, dehydroascorbate reductase and glutathione peroxidase, commonly reported to buffer oxidative stress in plants and algae, were not detected in E. compressa from any of the studied sites. Levels of total glutathione and phenolic compounds decreased in algae from mine‐impacted sites. In contrast, high levels of dehydroascorbate were found in algae from impacted sites, whereas ascorbate remained unchanged. Therefore, it is suggested that E. compressa tolerates a copper‐enriched environment, and the accompanying oxidative stress, through the accumulation of copper, activation of ascorbate peroxidase, synthesis of ascorbate (accumulated as dehydroascorbate) and consumption of glutathione and water‐soluble phenolic compounds.     

High levels of multiple metal resistance and its correlation to antibiotic resistance in environmental isolates of Acinetobacter

Forty strains of Acinetobacter were isolated from different environmental sources. All the strains were classified into four genospecies, i.e. A. baumannii (33 isolates), A. calcoaceticus (three isolates), A. junii (three isolates) and A. genospecies3 (one isolate). Susceptibility of these 40 strains to salts of 20 heavy metals and 18 antibiotics was tested by the agar dilution method. All environmental isolates of Acinetobacter were resistant to multiple metal ions (minimum 13 metal ions) while all but one of the strains were resistant to multiple antibiotics (minimum four antibiotics). The maximum number of strains were found to be sensitive to mercury (60% strains) while all strains were resistant to copper, lead, boron and tungsten even at 10 mm concentration. Salts of these four metal ions may be added to the growth medium to facilitate selective isolation of Acinetobacter. Rifampicin and nalidixic acid were the most toxic antibiotics, inhibiting 94.5 and 89.5% of the acinetobacters, respectively. A. genospecies3 was found to be the most resistant species, tolerating high concentrations of all the 20 metal ions and also to a greater number of antibiotics than any other species of Acinetobacter tested. An inhibitory concentration (10 mm) of Ni²⁺ and Zn²⁺ was observed to inhibit the growth of all of the clinical isolates but allowed the growth of the environmental isolates, facilitating the differentiation between pathogenic and non-pathogenic acinetobacters.