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PubMed2003

Detection and quantitative estimation of Dehalococcoides spp. in a dechlorinating bioreactor by a combination of fluorescent in situ hybridisation (FISH) and kinetic analysis.

The unique capacity of Dehalococcoides ethenogenes of completely dechlorinating the common groundwater pollutant tetrachloroethene (PCE) to the harmless ethene makes this microorganism very attractive for application in natural or engineered bioremediation systems. In this study, the qualitative and quantitative determination of Dehalococcoides spp. in a lab-scale bioreactor was performed based on the combination of fluorescent in situ hybridisation (FISH) for specific detection, and kinetic batch tests at non-limiting hydrogen and PCE concentration for quantitative determination. The dechlorinating bioreactor was operated at a high and constant PCE loading rate of 255 micromol PCE [g volatile suspended solids (VSS)](-1) day(-1). Pale coccoid cells resembling the distinctive morphotype of D. ethenogenes were present in the microbial culture. These cocci hybridised with both eubacterial probes and the Dhe1259t probe recently designed for detecting Dehalococcoides spp. Positive hybridisation was also observed when the DHC1377 reverse primer was used as a specific probe and applied to the dechlorinating microbial consortium. The maximum dechlorination rate obtained under non-limiting hydrogen and PCE concentrations was 3.22 +/- 0.08 mmol Cl(-) l(-1 )day(-1). From the specific activity of D. ethenogenes [i.e. 0.055 +/- 0.008 mmol Cl(-) (mg VSS)(-1) day(-1)], as reported from pure culture study, this observed maximum rate corresponded to a concentration of this bacterium in the mixed liquor of the bioreactor of 59.0+/-10.4 mg VSS.l(-1) (41.5+/-11.2% of overall VSS). This calculated relative abundance of D. ethenogenes was in agreement with the percentage of methanol (in terms of reducing equivalents) channeled to reductive dechlorination (approximately 30%) supporting the assumption that most reductive dechlorination was actually due to this microorganism.

Biodegradation, Environmental↗
PubMed2004

The generation of high biomass from chlororespiring bacteria using a continuous fed-batch bioreactor.

A continuous fed-batch reactor system was developed to rapidly obtain dense chlororespiring cultures of Anaeromyxobacter dehalogenans strain 2CP-C. A syringe pump continuously delivered concentrated 2,6-dichlorophenol (50-150 mM) to an anaerobic reactor vessel at a rate that sustained linear growth but prevented the substrate toxicity of chlorophenol. Dechlorination was not significantly inhibited by end product phenol up to 8 mM. A cell density of 76.8 mg protein l(-1) was obtained in 24 h. Specific growth rates averaged 0.033 h(-1)at 50% substrate limitation, which was in agreement with the maximum specific growth rate of 0.068 h(-1). This reactor system provides an efficient, cost-effective, and convenient method to rapidly obtain dense dechlorinating biomass and is promising to accelerate investigations of enzymes involved in chlororespiration.

Anaerobiosis↗
PubMed2006

Measurement of chlorine dioxide penetration in dairy process pipe biofilms during disinfection.

Biofilms are considered a significant health risk in the food and dairy industries because they can harbor pathogens, and direct contact with them can lead to food contamination. Biofilm control is often performed using strong oxidizing agents like chlorine and peracetic acid. Although chlorine dioxide (ClO2) is being used increasingly to control microbiological growth in a number of different industries, not much is known about disinfection in biofilms using chlorine dioxide. In this study, a microelectrode originally made for chlorine detection was modified to measure the profiles of chlorine dioxide in biofilm as a function of depth into the biofilm. In addition, discarded microelectrodes proved useful for in situ direct measurement of biofilm thicknesses. The chlorine dioxide microelectrode had a linear response when calibrated up to a ClO2 concentration of 0.4 mM. ClO2 profiles showed depletion of disinfectant at 100 microm in the biofilm depth, indicating that ClO2 may not reach bacteria in a biofilm thicker than this using a 25 mg/l solution.

Biofilms↗
PubMed2001

8-Cl-adenosine is an active metabolite of 8-Cl-cAMP responsible for its in vitro antiproliferative effects on CHO mutants hypersensitive to cytostatic drugs.

8-Cl-cAMP has been undergoing clinical trials as a potential chemotherapy agent, but there is much discussion in the literature as to whether the active agent is 8-Cl-cAMP itself, or its major metabolite, 8-Cl-adenosine. 8-Cl-cAMP is susceptible to the action of serum enzymes such as phosphodiesterases, and its metabolism when administered to cancer patients raises questions as to the mechanism of action of 8-Cl-cAMP. The stability of 8-Cl-cAMP when incubated with serum, and the effects of both 8-Cl-cAMP and 8-Cl-adenosine on the proliferation of variant lines of CHO cells hypersensitive to 8-Cl-cAMP were investigated. A solid-phase extraction (SPE) purification protocol and the HPLC method previously developed were used to determine 8-Cl-cAMP and 8-Cl-adenosine. Heat treatment of serum inactivated the enzymes in the culture medium responsible for activating 8-Cl-cAMP. Under these conditions 8-Cl-cAMP remained stable and there were no traces of its metabolite, 8-Cl-adenosine. Cell culture experiments showed that 8-Cl-cAMP only affected cell growth in medium that contained untreated serum. In contrast, 8-Cl-adenosine was shown to be growth inhibitory in medium containing either heat-treated or untreated serum. HPLC analysis of the culture medium from the cell culture experiments supported the hypothesis that 8-Cl-cAMP was only effective in inhibiting cell growth after metabolism to 8-Cl-adenosine. Thus further studies of this drug and its mechanism of action should focus on 8-Cl-adenosine.

2-Chloroadenosine↗
PubMed1997

Transformation of chlororesorcinol by the hydrocarbonoclastic yeasts Candida maltosa, Candida tropicalis, and Trichosporon oivide.

The inhibitory effects of chlorinated monoaromatic compounds on three hydrocarbonoclastic yeasts grown on glucose or resorcinol were examined. At concentrations of 1.0 M, all of the monoaromatic compounds were inhibitory. When the concentration of chlororesorcinol was significantly reduced (0.0005 M), the inhibition to each yeast was minimized. Extracts of the cultures of yeasts growing on resorcinol plus chlororesorcinol were analyzed for residual resorcinol and chlororesorcinol with high pressure liquid chromatography. Neither compound was detected in the culture broth of Candida maltosa, but several unidentified compounds were present. Only chlororesorcinol was detected in the culture broth of Trichosporon oivide, and both resorcinol and chlororesorcinol were present in extract from cultures of C. tropicalis. Cultures of C. maltosa grown on resorcinol-yeast nitrogen base, washed and suspended in phosphate buffer and subsequently incubated with chlororesorcinol, turned the culture broth a distinct pink color. The data indicate that C. maltosa has the potential to co-metabolize chlororesorcinol.

Biotransformation↗
PubMed2006

Biodegradation of 2,4,5-trichlorophenol by aerobic microbial communities: biorecalcitrance, inhibition, and adaptation.

Chlorinated aromatic compounds challenge our environment and wastewater treatment processes due to their biorecalcitrance and inhibition. In particular, 2,4,5-trichlorophenol (TCP) seems to demonstrate greater resistance to biodegradation than other trichlorophenols and is a known uncoupler of the electron transport chain, although little work addresses this compound specifically. Here, we investigate the biorecalcitrance, inhibition, and adaptation to 2,4,5-trichlorophenol by aerobic mixed microbial communities. We show that 2,4,5-trichlorophenol is strongly resistant to biodegradation at concentrations greater than 40 microM, demonstrates inhibition to respiration in direct proportion to 2,4,5-trichlorophenol concentration (with 50% inhibition projected near 85 microM 2,4,5-trichlorophenol), and does not sustain biomass in continuous reactors, even when all input 2,4,5-trichlorophenol is degraded. Communities showed consistent adaptation patterns to 2,4,5-trichlorophenol at concentrations of 10 microM and 20 microM, but these patterns diverged at concentrations greater than 40 microM. Finally, thermodynamic approximations were used to estimate the yield of 2,4,5-trichlorophenol as 0.165 gVSS/gCOD, a low value that partially explains why biodegradation of 2,4,5-trichlorophenol did not sustain the biomass. In particular, we estimated that the minimum concentration to support steady-state biomass (S (min)) is approximately 180 microM, a value much larger than the 40-microM concentration that is strongly resistant to biodegradation. Thus, readily biodegradable concentrations of 2,4,5-trichlorophenol are too low to sustain the biomass that biodegrades it.

Adaptation, Physiological↗
PubMed2006

Chemical and biological evaluation of sediments from the Wadden Sea, The Netherlands.

We describe the results of an evaluation of marine sediments using chemical measurements and bioassays. Four groups of chemicals, i.e., heavy metals, PAHs, chlorinated aromatic compounds, and tin compounds, were measured at 16 locations in the Wadden Sea, The Netherlands. Extractions of sediments from each location also were assessed using five bioassays. Our objective was to identify chemicals likely to pose biological risks, characterize the relation between bioassay results and particular classes of chemical(s) and determine "clean" reference sites on the basis of the chemical and biological evaluations. A multivariate technique (Principal Component Analysis; PCA) was used to meet these objectives. Results of the PCA indicated that the response of the Microtox Solid Phase bioassay had a positive, significant relationship with the levels of PAHs and organotin compounds. The responses of the other bioassays did not consistently relate to the concentrations of the other measured chemicals. Our findings indicate that the organotin compounds may still be a stressor for aquatic invertebrates in the Dutch Wadden Sea. On the basis of the chemical and biological evaluations, four sites (Dantziggat, Malzwin, Richel and Lauwers) can be considered to be "clean" reference sites.

Biological Assay↗
PubMed1988

Drug-protein conjugates--XIV. Mechanisms of formation of protein-arylating intermediates from amodiaquine, a myelotoxin and hepatotoxin in man.

The enzymic and non-enzymic formation of protein-arylating intermediates from amodiaquine (AQ,7-chloro-4-(3'-diethylamino-4'-hydroxyanilino) quinoline), an anti-malarial associated with agranulocytosis and liver damage in man, was studied in vitro. [14C]AQ in phosphate buffer, pH 7.4, under air was autoxidized to a reactive derivative(s) which possessed characteristics indicative of a semiquinone/quinone imine: reduction by NADPH and ascorbic acid, conjugation with thiols and irreversible binding to microsomal and soluble proteins. Cysteinyl SH groups were major sites of arylation. Radiolabelled material irreversibly bound to HSA after 24 hr and to human liver microsomes after 4 hr represented 26.5 +/- 1.8% and 31.4 +/- 0.6% (means +/- SD, N = 3) of incubated [14C]AQ (10 microM), respectively. The quinone imine of AQ(AQQI) was synthesized, and displayed the same oxidative and electrophilic reactions as the product(s) of AQ's autoxidation. A water-soluble product formed in buffered solutions of AQ and N-acetylcysteine was identified as an AQ mercapturate by comparison with an adduct prepared from synthetic AQQI. Irreversible binding of [14C]AQ was inhibited by a radical scavenger; this indicated that the semiquinone imine contributed to the binding. Although AQ was extensively de-ethylated by human liver microsomes, oxidation by cytochrome P-450 did not appear to be principally responsible for its activation and irreversible binding in microsomal incubations. AQ was oxidized to protein-arylating intermediates by horseradish peroxidase. It also formed reactive derivatives, possibly N-chloro compounds, in chlorine solutions. These findings indicated that AQ can give rise to chemically reactive species by at least three distinct mechanisms, viz. autoxidation in neutral solution under air, peroxidase-catalyzed oxidation and N-chlorination. Formation of such species in liver and myeloid cells might be responsible for the adverse reactions associated with AQ.

Adolescent↗
PubMed1993

Organic micropollutants in Swiss sewage sludge.

Sludges from sixteen municipal and eleven industrial Swiss sewage treatment plants were analysed for adsorbable organic compounds (AOX), chlorinated pesticides, polychlorinated biphenyls and polycyclic aromatic hydrocarbons. Except for the AOX values, which were significantly higher (p < 0.05) in industrial sludges, there was no significant difference between industrial and municipal sludges. The AOX values did not correlate with any of the measured pollutants. The methods employed (capillary GC-MS and GC-electron-capture detection) proved to be suitable for monitoring organic micropollutants in industrial and municipal sewage sludges.

Environmental Monitoring↗
PubMed1989

The effectiveness of two disinfectants on denture base acrylic resin with an organic load.

This study compared the biocidal effectiveness of chlorine dioxide and 5.25% sodium hypochlorite (diluted 1:10) on acrylic resin strips inoculated with Staphylococcus aureus, Candida albicans, or Escherichia coli in the presence of an organic load. Sterile acrylic resin strips were immersed in a solution containing 10% horse serum and 10(5) to 10(7) organisms/ml for each type of organism, then disinfected in chlorine dioxide, sodium hypochlorite, or 0.9 sterile saline for 30 seconds or 1, 2, or 4 minutes. After disinfection, the strips were neutralized and incubated for 72 hours. The results showed a difference between the ability of chlorine dioxide and sodium hypochlorite to kill the test organisms on acrylic resin strips when organic matter is present. Chlorine dioxide achieved complete disinfection of all three organisms within 2 minutes. Sodium hypochlorite achieved complete disinfection of all three organisms within 4 minutes.

Bacteria↗
PubMed1993

Practical denture disinfection.

Dental laboratory personel are at risk of contracting infections from dental prostheses that have not been properly disinfected. A 4% chlorhexidine scrub for 15 seconds followed by a 3-minute contact time with a chlorine dioxide solution was effective in disinfecting contaminated dentures. Chair-side disinfection of dental prostheses before laboratory procedures is the key to keeping microbial contamination out of the dental laboratory.

Chlorhexidine↗
PubMed1985

Evaluation of different treatment processes with respect to mutagenic activity in drinking water.

Treatment processes which are applied in The Netherlands during the preparation of drinking water have been evaluated with regard to introduction and removal of organic mutagens as well as halogenated organics. It appeared that the most efficient processes in reducing mutagenic activity were activated carbon filtration and artificial dune recharge. In general these processes were also the most efficient in removing halogenated organics. Using low doses of chlorine dioxide (less than 1 mg C1O2/l) for safety disinfection of drinking water, no change or substantial less mutagenic activity than by chlorination (1 mg Cl/l) was found. This counts too for the formation of halogenated organics. Transport chlorination of stored river Meuse water was able to introduce or activate mutagenic nitro organics which have not been found previously. Ozone treatment under field conditions showed mostly a tendency to decrease the activity of organic mutagens. It was also shown that dependent on the water quality and treatment conditions a slight increase of mutagenic activity occurred, but this activity would be reduced by increasing the ozone dose. It seems possible to optimize the ozone treatment conditions regarding the level of ozone dose and the contact time to avoid an increase of mutagenic activity. Furthermore it was shown that when a mutagenic raw water source was used a proper combination of treatment processes is able to produce drinking water in which no mutagenic activity could be detected under the test conditions. Finally it is stated that before far-reaching decisions with respect to use mutagenicity data for a selection of water sources or treatment processes will be made, more information on the relation mutagenic activity from drinking water and effects on human health should become available.

Animals↗
PubMed1985

Alternative methods for chlorination.

Existing disinfectants are oxidative agents which all present negative effects on subsequent treatment processes. None of them has decisive advantages over chlorine, although chlorine-dioxide and chloramines might at times be preferable. Optimum treatment practices will improve the removal of organic precursors before final disinfection which could then consist in a light chlorine addition. A philosophy of radical change in water treatment technology encompassing physical treatment without chemicals such as membrane filtration, solid disinfectants is presented.

Animals↗
PubMed1986

Tracers of sewage sludge in the marine environment: a review.

A review of the available and potential tracers of the movement and distribution of sewage sludge ('sludge') in and on marine sediments is presented. A brief review of 'active' sludge tracers is followed by a more extensive review of 'passive' sludge tracers. These can be broken down into five categories: Natural organic components of sludge--organic carbon, faecal steroids, carbohydrate/TOC ratio, tomato seeds. Synthetic organic compounds--silicones, chlorinated hydrocarbons, non-ionic surfactants and their degradation products, linear alkylbenzenes. Stable isotopes of carbon, nitrogen, hydrogen and sulphur. Inorganic substances and their properties--minerals, paint pigments, trace elements, magnetic properties. Biological/Microbiological--faecal bacteria, Clostridium welchii spores, amoebae, viruses, nitrogen-fixing enterobacteriaceae. A wide variety of sludge tracers are available with the choice of suitable tracers in an individual situation governed by the available equipment and expertise, cost considerations, ease of sampling requirements and determination, and the nature of other pollutant inputs in the vicinity of the input of interest. Ideally, the sludge tracers chosen should be unique to the input of interest, but this will rarely be achievable in practice. In areas of gross sludge contamination, a marine geophysical survey technique can be used to delineate the area affected and this provides a rapid cost-effective method for large areas.

Amoeba↗
PubMed1991

Chlorine dioxide as a post-disinfectant for Dutch drinking water.

Chlorine dioxide has some important advantages over chlorine with respect to water quality (no formation of trihalomethanes, no impairment of taste and no odor) and stability when used for oxidation/disinfection of drinking water. In this paper, results are presented of experiments into the consumption and reaction kinetics of chlorine dioxide in a number of (drinking) waters in The Netherlands. It was found that chlorine dioxide consumption is related to the dissolved oxygen content (DOC) of the water and the reaction time. Water samples from a plant that applied ozonation and activated carbon filtration had a very low chlorine dioxide consumption. Other water quality parameters, including pH and CO3(2-), did not have any influence on consumption. The temporary advised Dutch guidelines of 0.2 mg l-1 (dosage) is sufficient for activated carbon treated water. For other Dutch drinking waters, however, none of the 0.2 mg l-1 chlorine dioxide remained after a reaction time of 10 min, as was also found for the water of Dutch pumping stations where chlorine dioxide is at present used for disinfection.

Chlorine↗
PubMed1977

Halogenated hydrocarbons in the atmosphere.

The sources, sinks, and tropospheric abundancies of the gaseous halocarbons, so far as they are known, are listed and discussed. The relative importance of natural and of man-made halocarbons is discussed within the context of contemporary concern about the depletion of stratospheric ozone by chlorine-bearing compounds.

Atmosphere↗