• Journal of Applied Biological Chemistry
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• v.63 no.2
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• pp.125-129
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• 2020

Environmental pollution caused by various heavy metals is a serious global problem. To solve this problem, microbial bioremediation of contaminated metals has developed rapidly as an effective strategy when physical and chemical techniques are not suitable. In this study, cadmium (Cd)-tolerant soil bacteria were isolated via artificial induction in laboratory conditions instead of screening bacteria naturally adapted to metal-contaminated soils. Wild-type (WT) bacteria grown in uncontaminated soils were artificially and sequentially adapted to gradually increasing Cd concentrations of up to 15 mM. The resultant cells, named Soil-CdR15, survived at a Cd concentration of 10 mM, whereas WT cells failed to survive with 4 mM Cd on solid media for 2 d. In liquid media containing Cd, the SoilCdR15 cells grew with 15 mM Cd for 7 d, whereas the WT cells could not grow with 5 mM Cd. Both Soil-CdR15 and WT cells removed approximately 35% of Cd at the same capacity from liquid media containing either 0.5 or 1.0 mM Cd over 2 d. In addition to Cd, the Soil-CdR15 cells showed increased resistance to nickel, zinc, and arsenic compared to WT cells. The Soil-CdR cells were identified as Burkholderia sp. by partial sequencing of 16S rRNA. The data presented in this study demonstrate that isolation of heavy metal-tolerant microorganisms via artificial induction in laboratory conditions is possible and may be useful for the application of the microorganisms for the bioremediation of heavy metals.

• Journal of Microbiology and Biotechnology
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• v.30 no.7
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• pp.1005-1012
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• 2020

Acid mine drainage (AMD) has been a serious environmental issue that threatens soil and aquatic ecosystems. In this study, an acid-tolerant sulfate-reducing bacterium, strain S4, was isolated from the mud of an AMD storage pond in Vietnam via enrichment in anoxic mineral medium at pH 5. Comparative analyses of sequences of the 16S rRNA gene and dsrB gene involved in sulfate reduction revealed that the isolate belonged to the genus Desulfovibrio, and is most closely related to Desulfovibrio oxamicus (with 99% homology in 16S rDNA sequence and 98% homology in dsrB gene sequence). Denaturing gradient gel electrophoresis (DGGE) analyses of dsrB gene showed that strain S4 represented one of the two most abundant groups developed in the enrichment culture. Notably, strain S4 was capable of reducing sulfate in low pH environments (from 2 and above), and resistance to extremely high concentration of heavy metals (Fe 3,000 mg/l, Zn 100 mg/l, Cu 100 mg/l). In a batch incubation experiment in synthetic AMD with pH 3.5, strain S4 showed strong effects in facilitating growth of a neutrophilic, metal sensitive Desulfovibrio sp. strain SR4H, which was not capable of growing alone in such an environment. Thus, it is postulated that under extreme conditions such as an AMD environment, acid- and metal-tolerant sulfate-reducing bacteria (SRB)-like strain S4 would facilitate the growth of other widely distributed SRB by starting to reduce sulfate at low pH, thus increasing pH and lowering the metal concentration in the environment. Owing to such unique physiological characteristics, strain S4 shows great potential for application in sustainable remediation of AMD.

• Korean Journal of Environmental Agriculture
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• v.16 no.4
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• pp.311-319
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• 1997

Heavy metal-tolerant microorganisms, such as Pseudomonas putida, P. aeruginosa, P. chlororaphis and P. stutzeri which possessed the ability to accumulate cadmium, lead, zinc and copper, respectively, were isolated from industrial wastewaters and mine wasewaters polluted with various heavy metals. Metal binding sites in the cells were investigated by extracting the components of the cells through pretreatments with hot water, acid, alkli, chloroform-methanol or chloroform-methanol/concentrated alkali. The heavy metal accumulation was drastically decreased by pretreatment with alkali or chloroform-methanol/concentrated alkali, but the heavy metal accumulation was not changed by pretreatment with chloroform-methanol. The amount of heavy metal accumulation was remarkably decreased by decreasing crude protein remaining in the cell. These results suggested that proteins of cell components played an important role on the heavy metal accumulation.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1156-1167
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• 2018

The aim of this study was to isolate and characterize lead (Pb)-solubilizing bacteria from heavy metal-contaminated mine soils and to evaluate their inoculation effects on the growth and Pb absorption of Brassica juncea. The isolates were also evaluated for their plant growth-promoting characteristics as well as heavy metal and salt tolerance. A total of 171 Pb-tolerant isolates were identified, of which only 15 bacterial strains were able to produce clear haloes in solid medium containing PbO or $PbCO_3$, indicating Pb solubilization. All of these 15 strains were also able to dissolve the Pb minerals in a liquid medium, which was accompanied by significant decreases in pH values of the medium. Based on 16S rRNA gene sequence analysis, the Pb-solubilizing strains belonged to genera Bacillus, Paenibacillus, Brevibacterium, and Staphylococcus. A majority of the Pb-solubilizing strains were able to produce indole acetic acid and siderophores to different extents. Two of the Pb-solubilizing isolates were able to solubilize inorganic phosphate as well. Some of the strains displayed tolerance to different heavy metals and to salt stress and were able to grow in a wide pH range. Inoculation with two selected Pb-solubilizing and plant growth-promoting strains, (i.e., Brevibacterium frigoritolerans YSP40 and Bacillus paralicheniformis YSP151) and their consortium enhanced the growth and Pb uptake of B. juncea plants grown in a metal-contaminated soil. The bacterial strains isolated in this study are promising candidates to develop novel microbe-assisted phytoremediation strategies for metal-contaminated soils.

• Journal of Soil and Groundwater Environment
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• v.20 no.4
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• pp.113-121
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• 2015

The contaminated soil at abandoned smelter areas present challenge for remediation, as the degraded materials are typically deficient in nutrients, and rich in toxic heavy metals and metalloids. Bioremediation technique is to isolate new strains of microorganisms and develop successful protocols for reducing metal toxicity with heavy metal tolerant species. The present study collected metal contaminated soil and characterized for pH and EC values, and heavy metal contents. The pH value was 5.80, representing slightly acidic soil, and EC value was 13.47 mS/m. ICP-AES analytical results showed that the collected soil samples were highly contaminated with various heavy metals and metalloids such as lead (183.0 mg/kg), copper (98.6 mg/kg), zinc (91.6 mg/kg), and arsenic (48.1 mg/kg), respectively. In this study, a bacterial strain, Bacillus cereus KM-15, capable of adsorbing the heavy metals was isolated from the contaminated soils by selective enrichment and characterized to apply for the bioremediation. The effects of heavy metal on the growth of the Bacillus cereus KM-15 was determined in liquid cultures. The results showed that 100 mg/L arsenic, lead, and zinc did not affect the growth of KM-15, while the bacterial growth was strongly inhibited by copper at the same concentration. Further, the ability of the bacteria to adsorb heavy metals was evaluated.

• Korean Journal of Soil Science and Fertilizer
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• v.47 no.3
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• pp.213-216
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• 2014

Arsenic is a known hazardous metalloid not only to the animals but also to plants. With high concentrations, it can impede normal plant growth and cause even death of plants at extremely high levels. A known plant response to stress conditions such as toxic levels of metal (loids) is the production of stress ethylene, causing inhibitory effect on root growth in plants. When the effect of various arsenic concentrations was tested to maize plant, the stress ethylene emission proportionately increased with increasing concentration of As(V). The inoculation of two arsenic tolerant bacteria; Pseudomonas grimonti JS126 and Pseudomonas taiwanensis JS238 having respective high and low 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity reduced stress ethylene emission by 59% and 30% in maize grown in arsenic polluted soils. The result suggested the possible use of Pseudomonas grimonti JS126 for phytoremediation of arsenic polluted soils.

• Journal of Life Science
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• v.8 no.3
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• pp.339-347
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• 1998

As a part of a study on the development of microbiological methods for petroleum exploration, the distribution of the avarafe survival rate for heterotrophs to various heavy metal ions and hydrocarbons were surveyed and compared by the use of plate count method. In consequence of the avarage survival rate to heavy metal ions(2 hours treatment) and hydrocarbons(1 hour treatment) for heterotrophs isolatinf from soil samples(50cm depth) which located in Doum mountain(A, B and D site) and Aedowon(C site) at Pohang area, the survival rate of heterotrophs for nickel(600ppm), cobalt(500ppm), cadmiun(100ppm), mercury(20ppm), zinc(400 ppm) and lead(500ppm) were 73.7%, 82.6%, 76.8%, 9.5%, 77.8% and 73.6% at A site and 67.9%, 82.5%, 86.0%, 5.8%, 82.5% and 91.7% at B site, 87.8%, 79.8%, 87.5%, 7.0%, 84.2% AND 47.7% AT c SITE, AND 71.8%, 76%, 85.9%, 1,2%, 79.6% AND 88.3% AT D site, respectively. Also the survival rate of heterotrophs from A,B,C and D site to pentane and hexane(each concentration is 20%) were 26.7% and 42.5%, 11.8% and 8.1%, 44.3% and 36.2%, and 12% and 3.5%, respectively. therefore, heterotrophs from B and D site that alternated gravelstone, muddy sandstone and sandstone were higher survival rate to the heavy metal ions than heterotrophs from A site which mainly composed gravelstone. Also, heterotrophs from C site which mainly composed muddy sandstone and once produced natural gas were showed relatively higher survival rate to the heavy metal ions and hydrocarbons than the other sites. Consequently, we confirmed that the distributions of tolerant heterotrophs to heavy metal ions and hydrocarbons were differ from the lithological compositon.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.5
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• pp.817-821
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• 2012

Soil samples collected from abounded mines of Boryeong area in South Korea were used in isolating bacterial strains and their capacity to solubilize inorganic phosphates and heavy metal tolerance were assessed in vitro. Three different inorganic phosphate sources (Ca phosphate, Fe phosphate, and Al phosphate) and four different heavy metals (Co, Cd, Pb and Zn) each with three concentrations ($100{\mu}g\;mL^{-1}$, $200{\mu}g\;mL^{-1}$, and $400{\mu}g\;mL^{-1}$) were used. The bacterial isolates PSB-1, PSB-2, PSB-3, and PSB-4 solubilized significantly higher amount of Ca phosphate during the first five days of incubation though subsequent drop in soluble phosphorus level in the medium was observed at the later stage (after 5 days) of the incubation. Solubilization of Ca phosphate and Fe phosphate was concomitant with the acidification of the culture medium compared to the control where it remained constant. Isolated strains could solubilize Fe phosphate to certain extent ($25-45{\mu}g\;mL^{-1}$) though solubilization of Al phosphate was found negligible. All the isolates were tolerant to heavy metals (Cd, Pb, and Zn) up to the concentration of $400{\mu}g\;mL^{-1}$ except PSB-1 and PSB-8, which were shown to be vulnerable to Co even at $100{\mu}g\;mL^{-1}$. Heavy metal tolerant strains should be further evaluated for plant growth promoting activities also under field conditions in order to assess their agricultural and environmental significance.

• Ocean and Polar Research
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• v.25 no.1
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• pp.21-30
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• 2003

Seasonal fluctuations of physico-chemical and biological aspects of the environment were studied in Vladivostok harbour (Golden Horn Bay, the East Sea/Sea of Japan). The benthic community structure was described with a focus on size-spectra (bacteria, meio- and macrofauna) related with the chemical environment and chemical fluxes in sediment and to reveal their possible ecological role in the process of bioremediation of the environment. Samples from two sites with different concentrations of heavy metals (Fe, Zn, Cu, Pb, Mn, Cr, Ni Cd, Co) and petroleum hydrocarbon were assessed by a number of methods. These included plate counts of culturable bacteria, observation through a scanning electron (SEM) and transmission electron microscope (TEM). These approaches were complemented with microscopic assessments of the diversity of the benthic community. The specific communities had a limited number of species, tolerant to abnormally high levels of toxic compounds. The dominant species were presented by several sho.1-lived small polychaetes (Capitella capitata) and nematodes (Oncholaimium ramosum). The highest population density was recorded in microbenthos, in various diatoms, various physiological groups of bacteria which participate in biomineralization: marine heterotrophic bacteria, which oxidized oil, black oil in addition to groups resistant to heavy metals. They have the entire set of mechanisms for neutralizing the negative effect of those compounds, forming the detrital food web and biogeochemical circulation of material in sediments, which results in the biological self-recycling of sea basins. Macro- and meiobenthic organisms were more sensitive to a greater extent of $H_2S$ and petroleum hydrocarbons than to metal content, but the within-site rankings were the same as those achieved for microbiological analyses.

• Journal of Korean Society of Environmental Engineers
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• v.28 no.11
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• pp.1222-1227
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• 2006

In this study, nine strains were isolated from heavy metal-contaminated soil in a mine. The high efficiency bacteria, JH1, to be able removal cadmium and copper, was selected by the screen test. JH1 was identified as Ralstonia eutropha by 16S rDNA analysis, fatty acid analysis, and its morphological and biochemical characteristics. After the cadmium-contaminated soil was washed with citric acid solution(pH 6, 10 mM), Ralstonia eutropha JH1 was inoculated in the soil washing water. In order to determine the optimal cell concentration for inoculation, cell concentrations were considered in 0.5, 1.0, 2.0, 4.0 g/L, respectively. The removal efficiencies for cadmium in each cell concentration of Ralstonia eutropha JH1 were 49.9, 84.4, 89.7% and 89.9% of 110 mg/L(Cd), after 5 days culture in soil washing water. When Ralstonia eutropha JH1 was inoculated in soil washing water containing each cadmium(110 mg/L) and copper(100 mg/L), each of them was removed completely during 6 days culture. The completely removing time for cadmium and copper in each low concentration, 10, 30 and 60 mg/L were 12, 18 and 48 hrs, respectively.