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PubMed1985

Comparative study of membrane filtration and enrichment media for the isolation and enumeration of Pseudomonas aeruginosa from sewage, surface water, and swimming pools.

The recovery of Pseudomonas aeruginosa on several selective culture media was tested using raw sewage and secondary sewage effluent samples as well as spiked chlorinated imitation swimming water and samples from whirlpools. mPA-medium B gave good recovery of both vital and chlorine-injured P. aeruginosa and selectivity was greater than 90% when analysing whirlpool samples. It is therefore the medium recommended for examination of chlorinated swimming pools. When analysing sewage polluted water with the mPA-B medium, reduced selectivity was noted from low verification rates and from overgrowth by competitive flora. A modified medium (mPA-D; addition of cetrimide, omission of sulphapyridine and actidione) was more selective and sufficiently recovered noninjured cells. Chlorine-injured cells were completely inhibited, however. C-390 (9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan) was confirmed to be highly selective for P. aeruginosa when used in spread plates at a concentration of 30 micrograms/mL; P. aeruginosa was slightly inhibited. However, the medium could not be used with conventional membrane filtration techniques, because cellulose ester filters interfered with the selective action of C-390. Selectivity could be improved by using Gelman Tuffryn (polysulphone) filters and increasing the C-390 concentration to 120 micrograms/mL. At this concentration, however, the medium was strongly inhibitory to P. aeruginosa; resuscitation only partially improved recovery. Two other membrane filtration media were tested. Both cetrimide - nalidixic acid agar and Drake's medium No. 19 were inhibitory to chlorine-injured cells. Several types of membrane filters were tested and there was little difference between them.(ABSTRACT TRUNCATED AT 250 WORDS)

Cetrimonium Compounds↗

PubMed1998

A selective historical review of congener-specific human tissue measurements as sensitive and specific biomarkers of exposure to dioxins and related compounds.

Estimating internal exposure or dose of dioxins and related chemicals such as dibenzofurans and dioxinlike polychlorinated biphenyls is relatively straightforward in laboratory animals because a known dose is given and the amount absorbed can be measured. In wildlife, direct tissue measurement and measurement of environmental samples have both recently been used to estimate exposure. Until recently, human studies used only indirect indicators such as skin lesions to qualitatively estimate exposure to these chlorinated organic compounds. Environmental measurements have also sometimes been used to estimate human exposure. Dioxins in human tissue were not measured until the 1970s, when 2,3,7,8-tetrachlorodibenzo-p-dioxin was measured in mothers' milk; congener-specific measurement of dioxins and dibenzofurans in tissues (blood, milk, and adipose tissue) of the general population and exposed workers was first performed in the United States in the 1980s. Measurement in a sensitive and specific fashion of the 17 toxic dioxin and dibenzofuran congeners currently found in human tissue from industrial countries began in the 1980s. The use of known chemical standards, capillary columns, high resolution gas chromatography and mass spectrometry (GC-MS) has now become relatively common. GC-MS analysis of blood is currently accepted as the gold standard for estimating human exposure to dioxins. However, analyses are still costly and time consuming, and worldwide there are few qualified laboratories. There is currently a lack of knowledge concerning kinetics at higher and lower exposure levels for most of the toxic dioxin congeners and of levels in target tissues of concern.

Benzofurans↗

PubMed1988

Reduction of mutagenicity and toxicity of aflatoxin B1 by chlorine gas treatment.

Chlorine gas was used to treat aflatoxin B1 (AFB1). The time-related exposure study showed that 4 ml (15 mg) pure chlorine gas caused about 90% destruction of 100 micrograms AFB1 within 10 min, at standard temperature and pressure. Four fluorescent reaction products were produced, two of which were identified as 8,9-dichloro-AFB1 and 8,9-dihydroxy-AFB1 (diol). The use of [14C]AFB1 confirmed the 90% destruction of the compound by chlorine gas. An increased destruction of AFB1 also occurred when an increased amount of chlorine gas was used. The mutagenic activity of the AFB1 sample treated for 10 min was reduced to about 5% of the untreated control using the Salmonella typhimurium strain TA98 in the presence of a rat-liver S-9 mix. A similar time-related reduction in AFB1 toxicity after chlorine treatment was also achieved using the chicken embryo toxicity assay.

Aflatoxin B1↗

PubMed2001

Development of hexachlorobenzene-dechlorinating mixed cultures using polysorbate surfactants as a carbon source.

The use of three nonionic polysorbate surfactants--Tween 60, 61 or 65--as the sole carbon source to sustain methanogenesis and dechlorination, as well as the effect of long-term exposure of enriched cultures to these surfactants, was investigated through the development of three sediment-derived cultures. Over a one-year period, the carbon source in these cultures was gradually switched from glucose and methanol to surfactant only, while the surfactant concentration was increased from an initial concentration of 100 mg/L to 400 mg/L. In each feeding cycle, the surfactants were partially degraded and converted to methane. Transition from glucose to Tween surfactants as the electron donor did not affect the rate, extent, and pathway of HCB transformation. These surfactants sustained the reductive dechlorination of HCB even after one year of continuous addition to the enriched cultures. This study demonstrated that reductive dechlorination of HCB sustained by the fermentation of Tween surfactants is feasible. The results support the use of anaerobically degradable Tween surfactants for the biotransformation of polychlorinated organic compounds. In principle, these surfactants could be used to simultaneously increase the bioavailability of subsurface contaminants while serving as the carbon and electron source for microbial reductive dechlorination.

Biological Availability↗

PubMed1986

Rodent whole-embryo culture as a teratogen screening method.

Postimplantation embryo techniques can sensitively detect compounds with adverse biological activity. The versatility of the in vitro embryo toxicity/teratogenicity screening assay has been augmented by the addition of adult hepatic metabolic activation systems, which influence the results for some compounds in the embryo culture system. In this assay, cyclophosphamide produces negative results without metabolic activation and positive results with activation; metabolic activation changes the results for cytochalasin D from positive to negative. Another improvement has been the evaluation of the embryo toxicity/teratogenicity of various water-immiscible solvents. As these studies revealed that sonicated corn oil/serum preparations were nontoxic at concentrations of up to 10% of the culture medium, these solvents can now be used to expose cultured embryos to water-insoluble compounds (e.g., chlorinated pesticides). This in vitro rodent whole-embryo culture makes it possible to investigate extremely rapid cell division and specific time-related morphogenic events. Because in vitro development so closely parallels that occurring in vivo, the in vitro embryo culture system appears to be particularly relevant for the detection of teratogenic and embryotoxic compounds. A variety of compounds have been assayed in vitro via a combination of metabolic activation system and/or water-insoluble chemical delivery systems. Correlation between the published results of the in vitro screening assay and the published results of in vivo embryo toxicology and teratology studies has been very good. However, most of the compounds used in in vitro testing have been strongly teratogenic, and little information is available on compounds that are not teratogenic in in vivo tests. There is also a good correlation between the concentrations of teratogenic agents that cause adverse effects in vitro and the in vivo teratogenic doses of these compounds.

Animals↗

PubMed1985

Dichloromethane dehalogenase of Hyphomicrobium sp. strain DM2.

Dichloromethane dehalogenase, a highly inducible glutathione-dependent enzyme catalyzing the conversion of dichloromethane into formaldehyde and inorganic chloride, was purified fivefold with 60% yield from Hyphomicrobium sp. strain DM2. The electrophoretically homogeneous purified enzyme exhibited a specific activity of 17.3 mkat/kg of protein. Its pH optimum was 8.5. The enzyme was stable at -20 degrees C for at least 6 months. A subunit molecular weight of 33,000 was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gel filtration of native dichloromethane dehalogenase yielded a molecular weight of 195,000. Subunit cross-linking with dimethyl suberimidate confirmed the hexameric tertiary structure of the enzyme. Dichloromethane dehalogenase was highly specific for dihalomethanes. Its apparent Km values were 30 microM for CH2Cl2, 15 microM for CH2BrCl, 13 microM for CH2Br2, 5 microM for CH2I2, and 320 microM for glutathione. Several chlorinated aliphatic compounds inhibited the dichloromethane dehalogenase activity of the pure enzyme. The Ki values of the competitive inhibitors 1,2-dichloroethane and 1-chloropropane were 3 and 56 microM, respectively.

Bacteria↗

PubMed1975

[Effect of prothixen and related compounds on serotonin absorption by human thrombocytes].

Distribution of preparations in the order of reduction of the inhibitory influence on serotonin (ST) absorption by platelets corresponded to the distribution of the preparations, by decrease in the thymoanaleptic action: chlorimipramine greater than imipramine greater than amitryptiline desimpramine-nortyptyline-chlorprotixen greater than chlorpromaxine greater than norprotixen greater than protixen-propazine. A change in the influence of the preparation on the ST absorption by platelets caused by the administration into the structure of the chlorine atom compound or by demethylation of the preparation was largely determined by the chemical structure of the initial compound.

Absorption↗

PubMed1986

[Substances with antibacterial and antifungal activity. I. Synthesis and microbiologic activity of imidazolylmethylanilines].

The synthesis and antifungal activities of many derivatives of 1-(2-imidazolylmethyl)aniline and of 1-(4-imidazolylmethyl)aniline are reported. Antimicrobial data in comparison with miconazole show that many compounds containing chlorine atoms and nitro group exhibit an interesting antimycotic activity. The results obtained are discussed on the basis of structure-activity relationships.

Aniline Compounds↗

PubMed2006

CprK crystal structures reveal mechanism for transcriptional control of halorespiration.

Halorespiration is a bacterial respiratory process in which haloorganic compounds act as terminal electron acceptors. This process is controlled at transcriptional level by CprK, a member of the ubiquitous CRP-FNR family. Here we present the crystal structures of oxidized CprK in presence of the ligand ortho-chlorophenolacetic acid and of reduced CprK in absence of this ligand. These structures reveal that highly specific binding of chlorinated, rather than the corresponding non-chlorinated, phenolic compounds in the NH(2)-terminal beta-barrels causes reorientation of these domains with respect to the central alpha-helix at the dimer interface. Unexpectedly, the COOH-terminal DNA-binding domains dimerize in the non-DNA binding state. We postulate the ligand-induced conformational change allows formation of interdomain contacts that disrupt the DNA domain dimer interface and leads to repositioning of the helix-turn-helix motifs. These structures provide a structural framework for further studies on transcriptional control by CRP-FNR homologs in general and of halorespiration regulation by CprK in particular.

Bacterial Proteins↗

PubMed2001

Chlorin-type photosensitizers photochemically derived from vinyl porphyrins.

The reaction of singlet oxygen with four vinyl-substituted dicarboxylic porphyrins, vinyldeuteroporphyrin (VD), ethylvinyldeuteroporphyrin (EVD), hydroxyethylvinyldeuteroporphyrin (HVD) and protoporphyrin (PP) in organic solutions is investigated. The main products, the "hydroxyaldehyde" chlorin-type derivatives, are formed with a concentration-dependent photochemical quantum yield that reaches a maximum of 7.4 +/- 1.6 x 10(-3). However, owing to the high turnover of singlet-oxygen production, these chlorin-type compounds are easily prepared photochemically with a chemical yield of 70% and little side product formation. In chemical ionization mass spectrometry, these compounds display an unusual fragmentation with a loss of 16 mass units. This is attributed to the loss of the oxygen bound to the saturated carbon of the modified pyrrole unit. All these compounds sensitize the formation of singlet oxygen with a yield around 0.8. They interact with singlet oxygen with rate constants of 5 x 10(6)-9 x 10(6) M-1 s-1, lower than those measured for vinyl porphyrins. These data are likely to help in the characterization of photoproducts of vinyl porphyrins relevant to photodynamic therapy (PP, HVD). As exemplified with VD and EVD, they also point out the reaction of singlet oxygen as an efficient route to chlorin-type photosensitizers.

Chromatography, High Pressure Liquid↗

PubMed2003

Behaviour of Cd, Cr, Mn, Ni, Pb, and Zn in sewage sludge incineration by fluidised bed furnace.

Cd, Cr, Mn, Ni, Pb, and Zn behaviour during sewage sludge incineration was investigated in seven pilot tests using a circulating fluidised bed furnace. Dewatered sludge at a solids concentration of 15-18% was fed to the furnace either alone (two tests) or spiked with chlorinated organic compounds (five tests). The behaviour of metals in the fluidised bed furnace was studied by comparing metal concentrations in the two main ash streams: ash separated from the cyclone immediately following the fluidised bed furnace, and fly ash recovered in the final bag filter. A metal enrichment factor was defined as the ratio of metal concentration between filter ash and cyclone ash. Only Cd and Pb showed any significant enrichment. Their enrichment factors were mainly affected by chlorine concentration in the feed sludge. To check whether simple equilibrium models may explain and predict metal behaviour, experimental data were compared with percentage of the metal vaporisation in the combustion chamber predicted using a thermodynamic model. Discrepancies between model predictions and experimental results are accounted for by considering that kinetics may be a limiting factor in the formation of metal chloride gaseous species. Due to the very short sludge residence time in the fluidised bed furnace, the gaseous compounds have little chance to evolve completely.

Forecasting↗

PubMed2004

Behaviour of bisphenol A (BPA), 4-nonylphenol (4-NP) and 4-nonylphenol ethoxylates (4-NP1EO, 4-NP2EO) in oxidative water treatment processes.

Endocrine disrupting chemicals (EDCs) such as natural and synthetic hormones or industrial chemicals can adversely affect the endocrine system at very low concentrations. As such substances can be present in raw water used for drinking-water production, they potentially pose a health risk to humans. In this study laboratory tests were performed to determine removal efficiencies of selected oxidative drinking water treatment processes, namely ozonation (1.4 mg/l O3) and chlorination, using sodium hypochlorite (0.5 mg/l NaClO) and chlorine dioxide (0.4-0.6 mg/l ClO2) under conditions applied in technical plants. 500-300,000 ng/l of bisphenol A (BPA), 4-nonylphenol (4-NP) and 4-nonylphenol-n-ethoxylates (NPnEO) were selected for investigations and measured by HPLC/FLD and HPLC/MS. To investigate possible oxidation by-products, adsorbable organic halogens (AOX) were determined and estrogenic activities were assessed with the help of an estrogen receptor binding assay (YES). Ozonation and chlorination with ClO2 removed both 4-NP and BPA below detection limits, corresponding with AOX and estrogenic activity. Concerning NPnEO ozonation removed NP1EO and NP2EO up to 28% and 30%, respectively, whereas ClO2 showed high removal efficiencies, eliminating >94% and 92%, respectively. NaCIO removed 4-NP and BPA below detection limits, but estrogenic activities increased and AOX could be measured. NP1EO and NP2EO were only marginally reduced corresponding to the slight decrease of estrogenic potential.

Benzhydryl Compounds↗

PubMed1994

Identification of the bphA and bphB genes of Pseudomonas sp. strains KKS102 involved in degradation of biphenyl and polychlorinated biphenyls.

The nucleotide sequence of the upstream region of the bphC gene from Pseudomonas sp. strain KKS102 was determined. Four genes were found in this region. Deduced amino acid sequences of the first, second, third and fourth genes showed significant homology with a large subunit of iron-sulfur protein, a small subunit of iron-sulfur protein, ferredoxin and dihydrodiol dehydrogenase, respectively, from other bacteria which degrade biphenyl/polychlorinated biphenyls, toluene and benzene. E. coli, in which the four genes, bphC and the gene for ferredoxin reductase from benzene degrading bacterium were expressed, was able to produce meta-cleavage compounds from chlorinated biphenyls. These results show that these gene products are functional in both biphenyl and polychlorinated biphenyls degradation.

Alcohol Oxidoreductases↗

PubMed1995

Substrate diversity and expression of the 2,4,5-trichlorophenoxyacetic acid oxygenase from Burkholderia cepacia AC1100.

Burkholderia cepacia AC1100 uses the chlorinated aromatic compound 2,4,5-trichlorophenoxyacetic acid as a sole source of carbon and energy. The genes encoding the proteins involved in the first step (tftA and tftB [previously designated tftA1 and tftA2, respectively]) have been cloned and sequenced. The oxygenase, TftAB, is capable of converting not only 2,4,5-trichlorophenoxyacetic acid to 2,4,5-trichlorophenol but also a wide range of chlorinated aromatic phenoxyacetates to their corresponding phenolic derivatives, as shown by whole-cell and cell-free assays. The rate of substrate utilization by TftAB depends upon the extent of chlorination of the substrate, the positions of the chlorines, and the phenoxy group. These results indicate a mechanistic similarity between TftAB and the 2,4-dichlorophenoxyacetic acid/alpha-ketoglutarate-dependent dioxygenase, TfdA, from Alcaligenes eutrophus JMP134. The promoter of the oxygenase genes was localized by promoter-probe analysis, and the transcriptional start site was identified by primer extension. The beta-galactosidase activity of the construct containing the promoter region cloned upstream of the beta-galactosidase gene in the promoter-probe vector pKRZ-1 showed that this construct is constitutively expressed in Escherichia coli and in AC1100. The -35 and -10 regions of the oxygenase genes show significant sequence identity to typical Escherichia coli sigma 70 promoters.

Base Sequence↗

PubMed2000

Toluene-degrading bacteria are chemotactic towards the environmental pollutants benzene, toluene, and trichloroethylene.

The bioremediation of polluted groundwater and toxic waste sites requires that bacteria come into close physical contact with pollutants. This can be accomplished by chemotaxis. Five motile strains of bacteria that use five different pathways to degrade toluene were tested for their ability to detect and swim towards this pollutant. Three of the five strains (Pseudomonas putida F1, Ralstonia pickettii PKO1, and Burkholderia cepacia G4) were attracted to toluene. In each case, the response was dependent on induction by growth with toluene. Pseudomonas mendocina KR1 and P. putida PaW15 did not show a convincing response. The chemotactic responses of P. putida F1 to a variety of toxic aromatic hydrocarbons and chlorinated aliphatic compounds were examined. Compounds that are growth substrates for P. putida F1, including benzene and ethylbenzene, were chemoattractants. P. putida F1 was also attracted to trichloroethylene (TCE), which is not a growth substrate but is dechlorinated and detoxified by P. putida F1. Mutant strains of P. putida F1 that do not oxidize toluene were attracted to toluene, indicating that toluene itself and not a metabolite was the compound detected. The two-component response regulator pair TodS and TodT, which control expression of the toluene degradation genes in P. putida F1, were required for the response. This demonstration that soil bacteria can sense and swim towards the toxic compounds toluene, benzene, TCE, and related chemicals suggests that the introduction of chemotactic bacteria into selected polluted sites may accelerate bioremediation processes.

Alkanes↗

PubMedDate not supplied

[The gas chromatographic analysis of volatile compounds on the compact MKhP chromatograph].

Methods of analysis of biological specimens, alcohol beverages, and technological liquids in columns with standard adsorbents carbopaque B and C with carbowax 20M, widely used abroad, are described and examples of analyses presented. A special portable chromatographer (MCP) with flame ionization detector has been designed. It is intended for analysis of volatile organic compounds (alcohols, carbohydrates, organochlorine compounds, glycols, esters, etc.) in columns of different polarity. The system of processing of chromatographic findings permits a quantitative analysis of complex chromatograms and automated identification of substances in biological samples by using the available database.

Alcohols↗

PubMed2006

Coping with polychlorinated biphenyl (PCB) toxicity: Physiological and genome-wide responses of Burkholderia xenovorans LB400 to PCB-mediated stress.

The biodegradation of polychlorinated biphenyls (PCBs) relies on the ability of aerobic microorganisms such as Burkholderia xenovorans sp. LB400 to tolerate two potential modes of toxicity presented by PCB degradation: passive toxicity, as hydrophobic PCBs potentially disrupt membrane and protein function, and degradation-dependent toxicity from intermediates of incomplete degradation. We monitored the physiological characteristics and genome-wide expression patterns of LB400 in response to the presence of Aroclor 1242 (500 ppm) under low expression of the structural biphenyl pathway (succinate and benzoate growth) and under induction by biphenyl. We found no inhibition of growth or change in fatty acid profile due to PCBs under nondegrading conditions. Moreover, we observed no differential gene expression due to PCBs themselves. However, PCBs did have a slight effect on the biosurface area of LB400 cells and caused slight membrane separation. Upon activation of the biphenyl pathway, we found growth inhibition from PCBs beginning after exponential-phase growth suggestive of the accumulation of toxic compounds. Genome-wide expression profiling revealed 47 differentially expressed genes (0.56% of all genes) under these conditions. The biphenyl and catechol pathways were induced as expected, but the quinoprotein methanol metabolic pathway and a putative chloroacetaldehyde dehydrogenase were also highly expressed. As the latter protein is essential to conversion of toxic metabolites in dichloroethane degradation, it may play a similar role in the degradation of chlorinated aliphatic compounds resulting from PCB degradation.

Aroclors↗

PubMed2006

Comparison of reductive dechlorination of p-chlorophenol using Fe0 and nanosized Fe0.

Chlorophenols, as a kind of important contaminants in groundwater, are toxic and difficult to biodegrade. Laboratory tests were conducted to examine zero-valent iron as an enhancing agent in the dechlorination of chlorinated organic compounds. Nanoscale iron particles were synthesized from common precursors KBH(4) and FeSO(4). Batch experiments were performed to investigate the reduction of p-chlorophenol (4-CP) by both common Fe(0) and nanoscale Fe(0). Comparison of 300 mesh/100 mesh/commercial reductive iron powders showed that size of iron particles played an important role in reduction process. Initial concentration and pretreatment of iron particles also influenced the chlorination rate. Nanoscale Fe(0) offered much more advantages for treatment of 4-CP compared with common iron particles, such as stability and durability. And they can be used to treat contaminants in groundwater over a long time. Among different parts of synthesized nanoscale iron particle solution, the very fine particles were the major agent for treatment of pollutants. As for preservation of nanoscale Fe(0), ethanol was recommended.

Chlorine↗