Characterization of large plasmids encoding resistance to toxic heavy metals in Salmonella abortus equi

Salmonella abortus equi vaccine strains were found to be resistant to high levels of toxic heavy metals--arsenic, chromium, cadmium, and mercury. The two strains 157 and 158 were resistant to ampicillin also. Curing of these strains resulted in loss of one or more resistance marker indicating plasmid borne resistance. Plasmid profile of strain 157 showed presence of three plasmids of 85, 54, and 0.1 Kb, whereas 158 strain showed presence of 85 Kb and 2 Kb plasmids. Plasmids were isolated from strain 157 and introduced into E. coli DH5alpha with a transformation efficiency of 2 x 10(3) transformants/microg DNA. Interestingly the transformants were resistant to antibiotics, heavy metals (As, Cr, Cd, Hg) and was also able to utilize citrate, a trait specific to Salmonella species. We report and establish for the first time the transferable large plasmids encoding resistance to various heavy metals, antibiotics and biochemical nature of S. abortus equi.     

Sensitivity of thermophilic methanogenic bacteria to heavy metals

The effects of Cd, Cu, and Ni on pure cultures of thermophilic methanogenic bacteria were studied. The bacteria used wereMethanobacterium thermoautotrophicum and TAM, a thermophilic, acetate-decarboxylating, methanogenic bacterium. Much lower concentrations of heavy metals were needed to cause initial inhibition of TAM (1 mg/liter Cu and Cd; 5 mg/liter Ni) compared withM. thermoautotrophicum (10 mg/liter Cu and Cd; and 100 mg/liter Ni). No growth of TAM occurred at 5 mg/liter Cu and 25 mg/liter Ni, while the corresponding values forM. thermoautotrophicum were 50 mg/liter Cu and 200 mg/liter Ni. Cd (50 mg/liter) was totally inhibitory toM. thermoautotrophicum but allowed minimal growth of TAM. Ni stimulated both organisms at an optimal concentration of 5 mg/liter forM. thermoautotrophicum and 1 mg/liter for TAM. The toxicity of Cd and Cu was found to depend upon the presence of Ni in the medium.     

R-Bodies in newly isolated free-living hydrogen-oxidizing bacteria

R-Bodies have been found in a recently isolated pseudomonas-like free-living hydrogen oxidizing bacterium. Their isolation, fine structure and chemical composition are described and compared with the R-bodies from the kappa particles (Caedobacter), obligate endosymbionts of Paramecium aurelia. The 2K 1 R-bodies exhibited essential characteristics of the kappa R-bodies; however, their size and some other structural aspects proved that they represent a new type of R-bodies. The presence of phage tail-like particles in cells induced with Mitomycin C is in favour of the hypothesis that the R-bodies might be coded by defective prophages, or by extrachromosomal elements.     

Patterns of tolerance to heavy metals among methane-utilizing bacteria

The susceptibility of obligate methane-utilizing bacteria, including 14 reference strains and 175 environmental isolates, to five readily available heavy metal pollutants was determined. The chloride salts of Hg(II), Cd(II), Cu(II), Cr(III) and Zn(II) were tested in a system free from organic matter. Methane-utilizers appeared to have relatively discrete metal tolerance patterns with resistances to all metals varying with isolation site. Methanotrophs proved to be quite sensitive to mercury and cadmium but relatively resistant to copper, chromium and zinc     

Elemental analysis of uncultured magnetotactic bacteria exposed to heavy metals

Natural enrichments of magnetotactic bacteria were used to study the sites where heavy metals accumulate in uncultured bacteria. Most bacteria obtained by magnetic concentration from these enrichments contained, in addition to the magnetosomes, large phosphorus-rich granules in the cytoplasm. Metal (Zn, Mn, Sr, Cd, Al, Cr, and Pb) chlorides were added independently to the enrichments, and after 24 h, the elemental composition of the phosphorus-rich granules, magnetosomes, and "soft parts" (cytoplasm plus cell envelope) of whole bacteria was analyzed by energy-dispersive X-ray analysis on a transmission electron microscope. All bacteria contained Mn and Sr in the phosphorus-rich granules; some of them presented Mn peaks also in the soft parts. Zinc accumulation was variable and was found mainly in the phosphorus-rich granules, but also in the soft part of some bacteria. Some analyzed bacteria presented Zn peaks only in the soft parts, and some of them did not present Zn in any structure. Cadmium and Al were found only in the granules of some bacteria. Chromium was found in the soft parts of some bacteria. Lead was not detected in any bacteria. We concluded that the phosphorus-rich granules are major sites for metal accumulation by these bacteria. No conclusive results for magnetosomes were obtained because of the limitations of the analytical techniques particularly when used for whole cell analysis.     

Factors affecting immobilization of heavy metals by purple nonsulfur bacteria isolated from contaminated shrimp ponds

In order to remove heavy metals (HMs) from contaminated shrimp pond at the highest concentrations found of; 0.75 mg/l Cd²⁺, 62.63 mg/l Pb²⁺, 34.60 mg/l Cu²⁺ and 58.50 mg/l Zn²⁺, two strains of purple nonsulfur bacteria isolated from shrimp ponds (NW16 and KMS24) were investigated for their ability to immobilize HMs in 3% NaCl in both microaerobic-light and aerobic-dark conditions. Based on metabolic inhibition and metabolic-dependent studies, it was concluded that both strains removed HMs using biosorption and also bioaccumulation. The efficiency of removal by both strains with both incubating conditions tested was in the order of lead (Pb) > copper (Cu) > zinc (Zn) > cadmium (Cd). Optimal conditions for removal of HMs by strain NW16 were; cells in the log phase at 4.5 mg DCW/ml, pH 6.0, and 30°C for 30 min. With microaerobic-light conditions, the relative percent removal of HMs was: Pb, 83; Cu, 59; Zn, 39; Cd, 23 and slightly more with the aerobic-dark conditions (Pb, 90; Cu, 69; Zn, 46; Cd, 28). Cells in the log phase at 5.0 mg DCW/ml, pH 5.5, and 35°C for 45 min were optimal conditions for strain KMS24 and there were no significant differences for the removal percentages of HMs with either incubating conditions (averages: Pb, 96; Cu, 75; Zn, 46; Cd, 30). The presence of Ca²⁺ and Mg²⁺ significantly decreased the removal capacity of HMs for both strains.     

Magnetotactic bacteria: promising biosorbents for heavy metals

Magnetotactic bacteria (MTB), which can orient and migrate along a magnetic line of force due to intracellular nanosized magnetosomes, have been a subject of research in the medical field, in dating environmental changes, and in environmental remediation. This paper reviews the recent development of MTB as biosorbents for heavy metals. Ultrastructures and taxis of MTB are investigated. Adsorptions in systems of unitary and binary ions are highlighted, as well as adsorption conditions (temperature, pH value, biomass concentration, and pretreatments). The separation and desorption of MTB in magnetic separators are also discussed. A green method to produce metal nanoparticles is provided, and an energy-efficient way to recover precious metals is put forward during biosorption.     

香根草对土壤中几种重金属离子富集能力的比较研究

用组织培养的香根草无菌苗作为试验材料,进行香根草对Cd、Cr、Pb和Ni四种重金属离子的富集能力比较试验,分别测定了香根草体内的重金属含量、香根草与重金属元素的相关性、香根草对重金属的富集系数和根系滞留率.结果表明：香根草对四种重金属离子都具有较高的富集能力.其中,香根草植物体中元素总含量表现为Ni＞Cr＞Pb＞Cd,在茎叶和根中的分含量,均表现为根＞茎叶;香根草与重金属元素的关系中,Cd的相关性最显著;富集系数与重金属浓度呈负相关,以Ni最为明显;从滞留率的角度来看,以Cr在香根草根系的滞留率最大,但对同一种重金属来说,随浓度的变化,香根草根系对它的滞留率却几乎是恒定的.     

Bacterial resistance and detoxification of heavy metals

Microbial cells have resistances to essentially all of the toxic heavy metals of the Periodic Table. In bacterial cells, the genetic determinants of these resistances are frequently found on small extrachromosomal plasmids and transposons. Sometimes the resistances are associated with detoxifying enzymes. This is true for the Hg²⁺ → Hg⁰ reductase, the As³⁺ → As⁵⁺ oxidase and the Cr⁶⁺ → Cr³⁺ reductase. In other cases, such as As⁵⁺, Ag⁺ and Cd²⁺, no change in redox state occurs but, rather, uptake and transport differences accompany resistance determinants. This article summarizes what is known of bacterial metal resistances for which enzymatic detoxification is known to be the mechanism of resistance. The characteristics and functions of the enzymes are described, as well as a summary of the newer DNA sequence analysis (basic science) and bench-scale efforts (applied science) for the mercuric resistance system.     

Gene transfer in bacteria from soils contaminated with heavy metals

Transfer of metal resistance plasmids into two pseudomonad recipients, Pseudomonas aureofaciens and Ps. putida , from soil bacteria donor populations, was investigated in agricultural soil contaminated predominantly with Zn and Cu. The putative donor and recipient numbers on selective agar were not affected by the concentration of metals in the soils, nor were the number of transconjugants. However, there were differences in transfer frequencies of Hg and Cu resistance from the different soil samples. This is the first time that transfer of Cu resistance has been observed from native bacteria present in agricultural soils.     

Inhibition of methanogenesis by several heavy metals using pure cultures

The effect of different concentrations of nickel, copper and zinc on methanogenesis using pure cultures of Methanobacterium formicicum, Methanobrevibacter arboriphilicus, Methanosarcina thermophila and Methanospirillum hungatei over time (1, 15 and 30 d) was evaluated. methanobacterium formicicum showed the highest resistance to all the metals tested, while Methanospirillum hungatei was the most sensitive strain. All strains were sensitive to copper and zinc (10–250 mg 1^-1, but were much more resistant to nickel (200–1200 mg 1^-1). An adaptation process of the methanogenic pure culture with the toxicants was observed over time, which indicates that the inhibitory effects of heavy metals may be reverted in optimal anaerobic conditions.     

Anaerobic oxidation of alkanes by newly isolated denitrifying bacteria

The capacity of denitrifying bacteria for anaerobic utilization of saturated hydrocarbons (alkanes) was investigated with n-alkanes of various chain lengths and with crude oil in enrichment cultures containing nitrate as electron acceptor. Three distinct types of denitrifying bacteria were isolated in pure culture. A strain (HxN1) with oval-shaped, nonmotile cells originated from a denitrifying enrichment culture with crude oil and was isolated with n-hexane (C₆H₁₄). Another strain (OcN1) with slender, rod-shaped, motile cells was isolated from an enrichment culture with n-octane (C₈H₁₈). A third strain (HdN1) with oval, somewhat pleomorphic, partly motile cells originated from an enrichment culture with aliphatic mineral oil and was isolated with n-hexadecane (C₁₆H₃₄). Cells of hexane-utilizing strain HxN1 grew homogeneously in the growth medium and did not adhere to the alkane phase, in contrast to the two other strains. Quantification of substrate consumption and cell growth revealed the capacity for complete oxidation of alkanes under strictly anoxic conditions, with nitrate being reduced to dinitrogen. Received: 3 August / Accepted: 6 October 1999     

L-Phenylalanine formation from acetamidocinnamic acid by newly isolated bacteria

Summary Bacteria with the ability to form L-phenylalanine from acetamidocinnamic acid were isolated from several soils. Among them, strain no. S-7 and strain no. N-7 were identified as Alcaligenus faecalis S-7 and Bacillus sphaericus N-7, respectively. The L-phenylalanine-forming enzyme systems in both bacteria were found to be inducible and intracellular. With intact cells of both bacteria and 40 mg/ml as wet base, 10 mg/ml acetamidocinnamic acid was utilized, and 7.7 mg/ml L-phenylalanine in a molar yield of 94% was produced after 72h incubation. The pathway of L-phenylalanine formation is considered to take the following course: acetamidocinnamic acid is deacetylated to -amino cinnamic acid; this is spontaneously changed to phenylpyruvic acid, and L-phenylalanine is formed by transamination.     

植物修复重金属污染及内生细菌效应

土壤和水体的重金属污染已严重危害人类生存环境与健康。由于受重金属污染的环境分布广泛,迫切需要开发经济的清除环境重金属的技术。植物修复是通过绿色植物降解或移除环境污染物,有望成为重金属污染环境的原位修复技术。植物内生菌是指定殖于健康植物的各种组织和器官内部的细菌,被感染的宿主植物不表现出外在病症,耐重金属的内生菌在多种超富集植物中存在。在植物修复过程中,野生型内生菌或基因工程内生菌的抗性系统能降低重金属植物毒性,促进其迁移金属。耐重金属内生菌还可以通过固氮、溶解矿物元素及产生类植物激素、铁载体和ACC脱氨酶等产物促进植物的生长。主要综述目前植物-内生菌相互作用及其潜在的促进植物修复重金属污染的研究进展。     

外生菌根与植物抗重金属胁迫机理

外生菌根是外生菌根真菌和植物营养根形成的共生体,能够增加植物对污染胁迫的抵抗能力。本文综述了20多年来国内外研究外生菌根增加植物抗重金属毒害的成果,指出了外生菌根在植物抗重金属毒害中的积极作用,并概括其抗性的主要机理为：外延菌丝的吸收作用;菌根分泌物的调节与螯合作用;菌根菌套或哈蒂氏网吸收过滤有毒金属;菌根菌套的疏水性作用。在研究外生菌根抗重金属毒害机理的基础上,提出了该领域今后的研究前景。     

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.