胡杨根际细菌提高木本植物对重金属胁迫的耐受性

胡杨是我国西北荒漠地区特有的、对多种非生物逆境具有高抗逆性的树种,但其相关微生物的生态和生理功能研究还比较缺乏.本文从新疆沙雅地区原始胡杨林根际土壤中分离出重金属抗性细菌共72株.其中具有单一重金属(Cu²⁺、Ni²⁺、Pb²⁺或Zn²⁺)抗性的细菌菌株50株,有三重以上重金属抗性的菌株9株.将其中5株多重重金属抗性细菌接种至生根的竹柳插条,进行重金属胁迫下的盆栽培养.结果表明: 在铜或锌胁迫下,5株多重重金属抗性细菌对竹柳的生长抑制有不同程度的缓解,其中假单胞菌Z30和贪铜菌N8菌对铜和锌两种胁迫下竹柳生物量的增长与不接菌对照相比均达到显著差异水平.说明在非重金属污染区生长的胡杨根际存在多样性的重金属抗性细菌,其中一些多重重金属抗性菌对改善重金属胁迫下植物的生长有显著作用,具有应用于木本植物-微生物联合修复环境重金属污染的价值.     

Pollution,sources, and risks of heavy metals in coastal waters of China

Abstract

Comprehensive information on heavy metals in coastal waters at national scale of China is limited. Therefore, this study investigated the distribution, pollution, and ecological-health risks of heavy metals in coastal waters along 18,000?km coastline of China. Total 13 target heavy metals in coastal waters along coastline of China showed drastic spatial variations with average concentrations ranging from .14 (Cd) to 136.26 (Cu) μg/L. Cu was the dominant heavy metal with the maximal concentration of 1485.92?μg/L. Three methods including heavy metal pollution index (HPI), Nemerow index (NI), and contamination degree (CD) were adopted to explore heavy metal pollution. HPI obtained the worst-case evaluation results to illustrate that heavy pollution occurred at over 50% of sampling sites. Anthropogenic sources were the main sources of heavy metals in the coastal waters. Approximately 28.13% and 9.38% of sampling sites illustrated considerable and very high ecological risks, respectively. Metals including Cu, As, and Hg were the main pollution and risk contributors. Heavy metals in coastal waters posed high cancer risks and unacceptable non-cancer risks to both adults and children. Therefore, effective control of heavy metals is necessary for regional sustainability and well-beings of residents in coastal regions of China.     

Isolation of a naturally-occurring nickel resistance plasmid from a rare hypersaline estuary (Laguna Madre,Texas, USA) for potential use as a bio-indicator of metal contamination

Metal-resistant bacteria were isolated from sediments of the Laguna Madre, a rare hypersaline estuary impacted by many anthropogenic compounds, including various metals and metalloids. Bacteria were initially isolated on nutrient agar supplemented with NaCl; random isolates (n = 100) were tested for metal resistance toward zinc, nickel, chromium, and cadmium using a pour plate disc assay. Metal-resistant cultures were assayed for plasmids that contained naturally-occurring heavy metal resistance genes. Putative metal-resistance plasmids were tested for metal-resistance efficacy by transforming a metal-sensitive strain of Escherichia coli. Polymerase Chain Reaction (PCR) primers were designed to detect cnrA, part of a nickel–cobalt resistance gene cluster, and restriction endonuclease digests were performed to detect restriction sites within the plasmid. Results showed that many bacterial isolates tested were resistant toward most of the metals used in this study. Among tested bacteria cultures, 34 were resistant to zinc, 64 were resistant to chromium, and 51 resistant to cadmium. Only 8 cultures were resistant to nickel; however, most bacteria were found to be resistant to more than one metal. Several plasmids were found from the bacteria isolates. One plasmid, designated pDZ5, was isolated from a bacterium identified as Bacillus pumilus by 16S rRNA sequencing. Plasmid pDZ5 conferred nickel resistance to the metal-sensitive E. coli strain and was found to contain cnrA as confirmed by PCR amplification. Plasmid pDZ5 was successfully cut with restriction enzymes for potential ligation with reporter genes. The presence, abundance and expression of pDZ5 may prove to be a useful bio-indicator of metal contamination, specifically nickel pollution, in the Laguna Madre due to the fewer number of bacteria that were nickel-resistant compared to other metals.     

Potential of Siderophore Production by Bacteria Isolated from Heavy Metal: Polluted and Rhizosphere Soils

Recently, heavy metals have been shown to have a stimulating effect on siderophore biosynthesis in various bacteria. In addition, several studies have found that siderophore production is greater in bacteria isolated from soil near plant roots. The aim of this study was to compare the production of siderophores by bacterial strains isolated from heavy metal-contaminated and uncontaminated soils. Chrome azurol sulphonate was used to detect siderophore secretion by several bacterial strains isolated from heavy metal-contaminated and rhizosphere-uncontaminated soils with both a qualitative disc diffusion method and a quantitative ultraviolet spectrophotometric method. Siderophore production by rhizosphere bacteria was significantly greater than by bacteria isolated from contaminated soil. The Pearson’s correlation test indicated a positive correlation between the amount of siderophore produced by bacteria isolated from the rhizosphere using the quantitative and qualitative detection methods and the amount of heavy metal in the soil. However, a significant negative correlation was observed between the amount of siderophore produced by bacteria isolated from heavy metal-contaminated soil and the amount of heavy metal (r value of ?0.775, P < 0.001).     

Resistance to antibiotics and heavy metals of Pseudomonas aeruginosa isolated from natural waters

The resistance patterns of Pseudomonas aeruginosa strains isolated from freshwater and seawater to different antimicrobial agents and heavy metals were studied. It was found that resistance in these environments is linked to the level of faecal pollution. All strains were resistant to ampicillin and tetracycline but sensitive to carbenicillin, amikacin, tobramycin, and colistin, and to the assayed concentrations of silver, cadmium, lead, and molybdenum. Resistance to gentamicin, sulphadiazine, mercury, arsenic, and chromium was variable, so their use as epidemiological markers was considered. There was a close relationship between the degree of pollution and the frequency of heavy metal resistant strains of Ps. aeruginosa. The highest frequencies of resistance to mercury and arsenic were obtained from marine environments with little faecal pollution, where the highest incidence of multi-resistant strains was also observed.     

Resistance to antibiotics and heavy metals of Pseudontonas aeruginosa isolated from natural waters

The resistance patterns of Pseudomonas aeruginosa strains isolated from freshwater and seawater to different antimicrobial agents and heavy metals were studied. It was found that resistance in these environments is linked to the level of faecal pollution. All strains were resistant to ampicillin and tetracycline but sensitive to carbenicillin, amikacin, tobramycin, and colistin, and to the assayed concentrations of silver, cadmium, lead, and molybdenum. Resistance to gentamicin, sulphadiazine, mercury, arsenic, and chromium was variable, so their use as epidemiological markers was considered. There was a close relationship between the degree of pollution and the frequency of heavy metal resistant strains of Ps. aeruginosa. The highest frequencies of resistance to mercury and arsenic were obtained from marine environments with little faecal pollution, where the highest incidence of multi-resistant strains was also observed.     

Strain Polymorphism of the Plasmid Profiles in Acidithiobacillus ferrooxidans

Plasmid profiles were studied in 27 Acidithiobacillus ferrooxidans strains isolated from different geographic zones and substrates differing in composition of the main sulfide minerals, and also in experimentally obtained strains with acquired enhanced resistance to the ions of heavy metals (Fe, Ni, Cu, Zn, As). In 16 out of 20 strains isolated from different substrates, one to four 2- to 20-kb and larger plasmids were revealed. Plasmids were found in all five strains isolated from gold-containing pyrite–arsenopyrite ores and concentrates, in nine of 11 strains isolated from the ores and concentrates containing nonferrous metals, and in two of four strains isolated from the oxidation substrates of simple composition (mine waters, pyritized coals, active sludge). Changes in the plasmid profiles in some A. ferrooxidans strains (TFZ, TFI-Fe, TFV-1-Cu) with experimentally enhanced resistance to Zn²⁺, Fe³⁺, and Cu²⁺, respectively, were noted as compared with the initial strains. After 30 passages on a S⁰-containing medium, strain TFBk showed changes in the copy number of plasmids. The role of plasmids in the processes of oxidation of energy substrates and in the acquired enhanced resistance to heavy metal ions is discussed.     

From industrial sites to environmental applications with <Emphasis Type="Italic">Cupriavidus metallidurans</Emphasis>

Cupriavidus metallidurans CH34 and related strains are adapted to metal contaminated environments. A strong resistance to environmental stressors and adaptation make it ideal strains for survival in decreasing biodiversity conditions and for bioaugmentation purposes in environmental applications. The soil bacterium C. metallidurans is able to grow chemolithoautotrophically on hydrogen and carbon dioxide allowing a strong resilience under conditions lacking organic matter. The biofilm growth on soil particles allows coping with starvation or bad conditions of pH, temperature and pollutants. Its genomic capacity of two megaplasmids encoding several heavy metal resistance operons allowed growth in heavy metal contaminated habitats. In addition its specific siderophores seem to play a role in heavy metal sequestration besides their role in the management of bioavailable iron. Efflux ATPases and RND systems pump the metal cations to the membrane surface where polysaccharides serve as heavy metal binding and nucleation sites for crystallisation of metal carbonates. These polysaccharides contribute also to flotation under specific conditions in a soil-heavy metals–bacteria suspension mixture. An inoculated moving bed sand filter was constructed to treat heavy metal contaminated water and to remove the metals in the form of biomass mixed with metal carbonates. A membrane based contactor allowed to use the bacteria as well in a versatile wastewater treatment system and to grow homogeneously formed heavy metal carbonates. Its behaviour toward heavy metal binding and flotation was combined in a biometal sludge reactor to extract and separate heavy metals from metal contaminated soils. Finally its metal-induced heavy metal resistance allowed constructing whole cell heavy metal biosensors which, after contact with contaminated soil, waste, solids, minerals and ashes, were induced in function of the bioavailable concentration (Cd, Zn, Cu, Cr, Co, Ni, Tl, Pb and Hg) in the solids and allowed to investigate the speciation of immobilization of those metals. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Isolation and characterization of heavy metals resistant bacteria from Lagos Lagoon

A total of 228 bacteria with an ability to resist toxic heavy metals were isolated from 8 selected sites of the Lagos Lagoon. The bacteria isolated wereStaphylocaccus sp.,Bacillus sp.,Pseudomonas sp.,Streptococcus sp.,Moraxella sp.,Escherichia coli, Proteus sp.,Klebsiella sp. andSalmonella sp. The heavy metals to which resistance was recorded were mercury, lead, zinc, cobalt, copper and chromium. The lagoon sites from which the highest number of resistant bacteria were isolated were Marina and Ebute-Ero. The heavy metal to which most bacteria were resistant was cobalt, while the least was chromium. The significance of the result is discussed in relation to the Nigerian environment and human health.     

Complex interactions between diverse mobile genetic elements drive the evolution of metal-resistant bacterial genomes

In this study, we compared the genomes of three metal-resistant bacteria isolated from mercury-contaminated soil. We identified diverse and novel MGEs with evidence of multiple LGT events shaping their genomic structure and heavy metal resistance. Among the three metal-resistant strains, Sphingobium sp SA2 and Sphingopyxis sp SE2 were resistant to multiple metals including mercury, cadmium, copper, zinc and lead. Pseudoxanthomonas sp SE1 showed resistance to mercury only. Whole genome sequencing by Illumina and Oxford Nanopore technologies was undertaken to obtain comprehensive genomic data. The Sphingobium and Sphingopyxis strains contained multiple chromosomes and plasmids, whereas the Pseudoxanthomonas strain contained one circular chromosome. Consistent with their metal resistance profiles, the strains of Sphingobium and Sphingopyxis contained a higher quantity of diverse metal resistance genes across their chromosomes and plasmids compared to the single-metal resistant Pseudoxanthomonas SE1. In all three strains, metal resistance genes were principally associated with various novel MGEs including genomic islands (GIs), integrative conjugative elements (ICEs), transposons, insertion sequences (IS), recombinase in trio (RIT) elements and group II introns, indicating their importance in facilitating metal resistance adaptation in a contaminated environment. In the Pseudoxanthomonas strain, metal resistance regions were largely situated on a GI. The chromosomes of the strains of Sphingobium and Sphingopyxis contained multiple metal resistance regions, which were likely acquired by several GIs, ICEs, numerous IS elements, several Tn3 family transposons and RIT elements. Two of the plasmids of Sphingobium were impacted by Tn3 family transposons and ISs likely integrating metal resistance genes. The two plasmids of Sphingopyxis harboured transposons, IS elements, an RIT element and a group II intron. This study provides a comprehensive annotation of complex genomic regions of metal resistance associated with novel MGEs. It highlights the critical importance of LGT in the evolution of metal resistance of bacteria in contaminated environments.     

Characterization of aerobic polycyclic aromatic hydrocarbon-degrading bacteria from Bizerte lagoon sediments, Tunisia

Aims: To characterize polycyclic aromatic hydrocarbon (PAH)‐degrading bacteria from sediments of the Bizerte lagoon, and to determine their ability to resist other pollutants such as antibiotics and heavy metals. Methods and Results: More than 100 strains were isolated for their ability to use fluoranthene as the sole carbon and energy source. Most of them showed antibiotic and heavy metal resistance; 20 representative strains were selected for further analysis. 16S rRNA coding sequences analysis showed that the majority of the selected bacteria (75%) were affiliated to the Gammaproteobacteria. The selected strains also utilized high molecular weight PAHs containing up to four benzene rings and showed different profiles of PAH substrate usage suggesting different PAH degradation pathways. These results are consistent with the fact that nah‐like genes and idoA‐like genes, involved in PAH degradation, were detected in 6 and 1 strains respectively. Conclusions: The Bizerte lagoon, polluted by many human activities, leads to the co‐selection of strains able to cope with multiple contaminants. Significance and Impact of the Study: Polluted areas are often characterized by the concomitant presence of organic pollutants, heavy metals and antibiotics. This study is one of the first showing bacterial strains adapted to multiple contaminants, a promising potential for the development of bioremediation processes.     

New mobile genetic elements in <Emphasis Type="Italic">Cupriavidus metallidurans</Emphasis> CH34, their possible roles and occurrence in other bacteria

Cupriavidus metallidurans strain CH34 is a β-Proteobacterium that thrives in low concentrations of heavy metals. The genetic determinants of resistance to heavy metals are located on its two chromosomes, and are particularly abundant in the two megaplasmids, pMOL28 and pMOL30. We explored the involvement of mobile genetic elements in acquiring these and others traits that might be advantageous in this strain using genome comparison of Cupriavidus/Ralstonia strains and related β-Proteobacteria. At least eleven genomic islands were identified on the main replicon, three on pMOL28 and two on pMOL30. Multiple islands contained genes for heavy metal resistance or other genetic determinants putatively responding to harsh environmental conditions. However, cryptic elements also were noted. New mobile genetic elements (or variations of known ones) were identified through synteny analysis, allowing the detection of mobile genetic elements outside the bias of a selectable marker. Tn4371-like conjugative transposons involved in chemolithotrophy and degradation of aromatic compounds were identified in strain CH34, while similar elements involved in heavy metal resistance were found in Delftia acidovorans SPH-1 and Bordetella petrii DSM12804. We defined new transposons, viz., Tn6048 putatively involved in the response to heavy metals and Tn6050 carrying accessory genes not classically associated with transposons. Syntenic analysis also revealed new transposons carrying metal response genes in Burkholderia xenovorans LB400, and other bacteria. Finally, other putative mobile elements, which were previously unnoticed but apparently common in several bacteria, were also revealed. This was the case for triads of tyrosine-based site-specific recombinases and for an int gene paired with a putative repressor and associated with chromate resistance.     

Low occurrence of multi-antimicrobial and heavy metal resistance in Salmonella enterica from wild birds in the United States

Wild birds are common reservoirs of Salmonella enterica. Wild birds carrying resistant S. enterica may pose a risk to public health as they can spread the resistant bacteria across large spatial scales within a short time. Here, we whole-genome sequenced 375 S. enterica strains from wild birds collected in 41 U.S. states during 1978–2019 to examine bacterial resistance to antibiotics and heavy metals. We found that Typhimurium was the dominant S. enterica serovar, accounting for 68.3% (256/375) of the bird isolates. Furthermore, the proportions of the isolates identified as multi-antimicrobial resistant (multi-AMR: resistant to at least three antimicrobial classes) or multi-heavy metal resistant (multi-HMR: resistant to at least three heavy metals) were both 1.87% (7/375). Interestingly, all the multi-resistant S. enterica (n = 12) were isolated from water birds or raptors; none of them was isolated from songbirds. Plasmid profiling demonstrated that 75% (9/12) of the multi-resistant strains carried resistance plasmids. Our study indicates that wild birds do not serve as important reservoirs of multi-resistant S. enterica strains. Nonetheless, continuous surveillance for bacterial resistance in wild birds is necessary because the multi-resistant isolates identified in this study also showed close genetic relatedness with those from humans and domestic animals.     

Antibiotic and heavy metal resistance in Gram-negative bacteria isolated from the Seyhan Dam Lake and Seyhan River in Turkey

This study aimed to determine the pattern of antibiotic and heavy metal resistance in Gram-negative bacteria isolated from five different sites in the Seyhan Dam Lake and Seyhan River in Adana, Turkey. The susceptibility of 268 isolates to 16 different antibiotics and five heavy metals was investigated by agar diffusion and dilution methods, respectively. The most common species isolated from the samples were Aeromonas hydrophila (17.5 %), Aeromonas caviae (8.9 %) and Citrobacter freundii (8.9 %). There was a high incidence of resistance to ampicillin (80.2 %), streptomycin (71.6 %) and cefazolin (60.4 %). Multiple antibiotic resistance indices ranged from 0.2 to 0.81, suggesting exposure to antibiotic contamination. The isolates showed tolerance to different concentrations of heavy metals. These results indicate that antibiotic and heavy metal resistance among the Seyhan Dam Lake and Seyhan River bacteria may pose a risk to the fish population and public health. At the same time, the finding in the aquatic environments of different combinations of resistance genes suggests their involvement in the spread of multidrug-resistant strains.     

Metal and antibiotic resistance traits in Bradyrhizobium sp. (cajanus) isolated from soil receiving oil refinery wastewater

Forty isolates of Bradyrhizobium sp. (cajanus) were isolated from the nodules of pigeon pea plants grown in fields receiving petrochemical industrial wastewater for the past 12 years and characterized using standard methods. The heavy metal analysis of field soil and treated wastewater showed their presence in varying concentrations. All isolates showed resistance to one or more metals at concentrations >200 g/ml. Multiple metal resistance was a common phenomenon in these isolates. There was no correlation between extractable soil metal concentration and the ability of the isolates to tolerate metal salts in their growth medium as evidenced from their minimum inhibitory concentration (MIC). However, high incidence of metal resistance and the multiple nature of resistance might have been the result of continuous exposure of these strains to heavy metals in the treated wastewater of Mathura Oil Refinery. These strains were also found to be resistant to one or more of the 13 antibacterial drugs tested.     

龙须菜几株附生细菌对铜离子吸附性能的比较

重金属污染和富营养化是沿岸海域的主要污染形式, 造成沿海生态功能的退化。在重金属污染区域往往伴随着富营养化现象。本文对分离来自电镀废水中铜污染海域内的龙须菜体表的附生微生物对Cu2+的生物吸附进行试验, 分析和比较这些微生物对Cu2+的生物吸收性能。共分离到6株优势附生细菌：人苍白杆菌GF-S1、杀鲑气单胞菌GF-S2、莱拉微球菌GF-S3、纹带棒杆菌GF-S4、假交替单胞菌GF-S5和弗氏弧菌GF-S6。经对Cu2+的生物吸附试验发现, GF-S2效果最好, GF-S6最差, 其余间于这两者之间。对GF-S2、GF-S4和GF-S6进行了最佳吸附时间和pH筛选, 以及使用不同化学试剂对细菌Cu2+吸附的影响试验。结果显示：最佳吸附时间在40 min~60 min, pH介于4~5对GF-S2、GF-S4吸附Cu2+的效果较好; HCl处理导致GF-S2丧失吸附能力, NaOH和乙醇能提高其吸附性能; 3种试剂都能较大地提高GF-S4和GF-S6的吸附性能, 尤其对GF-S4经处理后, 对环境中的Cu2+的去除率得以大幅度提高, 去除较彻底。     

Effect of the Heavy Metals on Developmental Stages of Ovule,Pollen, and Root Proteins in <Emphasis Type="Italic">Reseda lutea</Emphasis> L. (Resedaceae)

Heavy metals are some of the most important environmental pollutants. Excessive amounts of heavy metals adversely affect plant growth and development. Also, the presence of elevated levels of heavy metal ions triggers a wide range of cellular responses including changes in gene expression and synthesis of metal-detoxifying peptides. The overall objective of this research was to elucidate some microscopic effects of heavy metals on the formation, development, and structure of pollen, ovule, and embryo and also root proteins in Reseda lutea L. For this purpose, the vicinity of Ahangaran lead–zinc mine (Hamedan, Iran) was chosen as a polluted area where amount of some heavy metals was several times higher than the natural soils. Flowers and young buds were collected from non-polluted and polluted plants, fixed in FAA₇₀, and studied during developmental stages by light microscopy. The results showed that heavy metals can cause some abnormalities during the pollen and ovule developmental process. The number of pollen grains was decreased, and their shape was changed. Increasing in thickness of the callosic wall and stabilizing of tapetum layer were observed in polluted plants. Asymmetrical formation of ovular integuments, degradation of egg apparatus, irregular formation of embryo sac, considerable vacuolation of embryonic cells, and degeneration of embryo in the late stage of heart-shaped embryo are the results of heavy metal pollution. For protein studies, young roots were harvested from plants exposed to pollution and non-exposed to pollution at the same time. Root proteins were extracted and studied by electrophoresis. The results revealed that some new proteins were synthesized in polluted samples that probably elevate plant tolerance to heavy metals.     

Strain polymorphism of the plasmid profiles in Acidithiobacillus ferrooxidans

Plasmid profiles were studied in 27 Acidithiobacillus ferrooxidans strains isolated from different geographic zones and substrates differing in the composition of the main sulfide minerals, and also in experimentally obtained strains with acquired enhanced resistance to the ions of heavy metals (Fe, Ni, Cu, Zn, As). In 16 out of 20 strains isolated from different substrates, one to four 2- to 20-kb and larger plasmids were revealed. Plasmids were found in all five strains isolated from gold-containing pyrite-arsenopyrite ores and concentrates, in nine of 11 strains isolated from the ores and concentrates containing nonferrous metals, and in two of four strains isolated from the oxidation substrates of simple composition (mine waters, pyritized coals, active sludge). Changes in the plasmid profiles in some A. ferrooxidans strains (TFZ, TFI-Fe, TFV-1-Cu) with experimentally enhanced resistance to Zn2+, Fe3+, and Cu2+, respectively, were noted as compared with the initial strains. After 30 passages on S0-containing medium, strain TFBk showed changes in the copy number of plasmids. The role of plasmids in the processes of oxidation of energy substrates and in the acquired enhanced resistance to the heavy metal ions is discussed.