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A H Neilson

Publications and source records attributed to A H Neilson.

18 recordsLinked to original sources
PubMed1994

Bioavailability of chlorocatechols in naturally contaminated sediment samples and of chloroguaiacols covalently bound to c(2)-guaiacyl residues.

Bacteria in anaerobic enrichment cultures that dechlorinated a range of chlorocatechols were used to examine the stability of endogenous chlorocatechols in a contaminated sediment sample and in interstitial water prepared from it. During incubation of the sediment sample for 450 days with or without added cells, there was a decrease in the concentration of solvent-extractable chlorocatechols but not in that of the total chlorocatechols, including sediment-associated components. In the presence of azide, the decrease in the concentrations of the former was eliminated or substantially decreased. Control experiments in which 3,4,5-trichlorocatechol was added to the sediment suspensions after 130 days showed that its dechlorination was accomplished not only by the added cells but also by the endemic microbial flora. It was concluded that the endogenous chlorocatechols in the sediment were not accessible to microorganisms with dechlorinating activity. On the other hand, microorganisms were apparently responsible for decreasing the solvent extractability of the chlorocatechols, and this effect decreased with increasing length of exposure time. Similar experiments carried out for 70 days with the sediment interstitial water showed that the chlorocatechols that were known to be associated with organic matter were also inaccessible to microbial dechlorination. Experiments with model compounds in which 4,5,6-trichloroguaiacol and tetrachloroguaiacol were covalently linked to C(2)-guaiacyl residues showed that these compounds were resistant to O demethylation or dechlorination during incubation with a culture having these activities. The only effect of microbial action was the quantitative reduction in 12 days of the C'1 keto group to an alcohol which was stable against further transformation for up to 65 days. The results of these experiments are consistent with the existence of chlorocatechols and chloroguaiacols in contaminated sediments and illustrate the cardinal significance of bioavailability in determining their recalcitrance to dechlorination and O demethylation, respectively. It is suggested that bioavailability is an important factor in determining the persistence of xenobiotics in natural ecosystems and that its omission represents a serious limitation in the interpretation of many laboratory experiments directed towards determining the persistence of xenobiotics in aquatic ecosystems.

Journal Article↗
PubMed1992

Role of sulfate concentration in dechlorination of 3,4,5-trichlorocatechol by stable enrichment cultures grown with coumarin and flavanone glycones and aglycones.

Metabolically stable anaerobic enrichment cultures have been obtained from sediment samples contaminated with chlorophenolic compounds. Enrichment was carried out with esculin, esculetin, naringin, naringenin, fraxin, quercetin, and acetate in media with two sulfate concentrations. These cultures were used to examine the O-demethylation of 4,5,6-trichloroguaiacol and the dechlorination of 3,4,5-trichlorocatechol. Whereas O-demethylation was observed in all cultures, the occurrence of dechlorination was significantly more restricted. The presence of the carbohydrate moiety in the cultures enriched with the glycones repressed development of populations which were able to carry out dechlorination. Although sulfate at a concentration of 2 g/liter in the primary enrichments blocked the development of populations able to bring about dechlorination, addition of sulfate at this concentration did not inhibit dechlorination in cultures possessing this capability. Different dichlorocatechol isomers were produced under the various conditions, so that in view of the established resistance of some of these to further dechlorination, the ultimate fate of 3,4,5-trichlorocatechol in the natural environment remains partly unresolved. No enrichment culture containing a low sulfate concentration was able to dechlorinate either 2,4,5-trichlorophenol or 2,4,6-trichlorobenzoate.

Anaerobiosis↗
PubMed1991

Chlorinated benzo-1,2-quinones: an example of chemical transformation of toxicants during tests with aquatic organisms.

An analytical procedure specific for chlorinated benzo-1,2-quinones has been developed to examine the stability of these compounds under conditions used for investigating their toxicity to aquatic organisms. Solutions of the compounds in a number of organic solvents were unstable in the light, and addition of acetone solutions to water brought about rapid decomposition of the chloroquinones which had half-lives less than 0.5 hr: the corresponding chlorocatechols were the principal products. The kinetics of decomposition of tetrachlorobenzo-1,2-quinone in aqueous solutions were studied in detail and showed the formation of tetrachlorocatechol, 2,5-dichloro-3,6-dihydroxybenzo-1,4-quinone, 1,2,3-trihydroxy-4,5,6-trichlorobenzene, 1,2,4-trihydroxy-3,5,6-trichlorobenzene, dichloromaleic acid, and a trichlorocyclopentendione. In organic solvents in the light, 3,4,5-trichlorobenzo-1,2-quinone underwent a dismutation reaction with formation of 3,4,5-trichloro- and tetrachlorocatechol; in a comparable reaction, 4,5-dichlorobenzo-1,2-quinone formed 4,5-dichlorocatechol and 3,4,5-trichlorocatechol. The toxicity of aqueous solutions prepared by dilution of freshly prepared acetone solutions of tetrachlorobenzo-1,2-quinone was examined in the zebra fish embryo/larvae test, and it was found that the threshold toxic concentration could be accounted for entirely by the analytically established concentration of tetrachlorocatechol produced as a chemical transformation product. It is concluded that in toxicological examination of reactive compounds, exposure to the toxicant should be assessed from concentrations analytically determined during the experiments and that attention be directed to both the nature and the toxicity of the transformation products.

Acetylation↗
PubMed1991

Dechlorination of chlorocatechols by stable enrichment cultures of anaerobic bacteria.

Metabolically stable anaerobic cultures obtained by enrichment with 5-bromovanillin, 5-chlorovanillin, catechin, and phloroglucinol were used to study dechlorination of chlorocatechols. A high degree of specificity in dechlorination was observed, and some chlorocatechols were appreciably more resistant to dechlorination than others: only 3,5-dichlorocatechol, 4,5-dichlorocatechol, 3,4,5-trichlorocatechol, and tetrachlorocatechol were dechlorinated, and not all of them were dechlorinated by the same consortium. 3,5-Dichlorocatechol produced 3-chlorocatechol, 4,5-dichlorocatechol produced 4-chlorocatechol, and 3,4,5-trichlorocatechol produced either 3,5-dichlorocatechol or 3,4-dichlorocatechol; tetrachlorocatechol produced only 3,4,6-trichlorocatechol. Incubation of uncontaminated sediments without additional carbon sources brought about dechlorination of 3,4,5-trichlorocatechol to 3,5-dichlorocatechol. O-demethylation of chloroguaiacols was generally accomplished by enrichment cultures, except that catechin enrichment was unable to O-demethylate tetrachloroguaiacol. None of the enrichments dechlorinated any of the polychlorinated phenols examined. The results suggested that dechlorination was not dependent on enrichment with or growth at the expense of chlorinated compounds and that it would be premature to formulate general rules for the structural dependence of the dechlorination reaction.

Journal Article↗
PubMed1990

Incorporation of a subacute test with zebra fish into a hierarchical system for evaluating the effect of toxicants in the aquatic environment.

Single-species laboratory tests were used to assess the acute toxicity of halogenated phenolic compounds. No single test system was most sensitive to all of the compounds examined, substantial variations in the sensitivity of the various organisms were noted, and there was no correlation between the toxicities assayed with different test systems. The zebra fish (Brachydanio rerio) embryo/larvae system was used to examine subacute effects using two of the compounds, and a protocol was developed with 6 weeks preexposure to the toxicant. Preexposure decreased the lowest observable effect concentration by a factor of about 4, and the effect was completely reversible during a 6-week postexposure period in the absence of the toxicant. An enclosed system for carrying out the zebra fish embryo/larvae test was developed and evaluated with three neutral volatile compounds: the median survival time and the frequency of occurrence of deformation were examined as end points. The effect of pH on toxicity was evaluated in buffered media for four of the test systems: toxicity increased markedly at the lower pH values, and it could be shown that the ionized forms of the phenols were not the only contributors to toxicity. It is proposed that the zebra fish system incorporating preexposure could be incorporated into a hierarchical system using a range of organisms for assessing acute toxicity in single species under laboratory conditions and multicomponent systems simulating natural ecosystems.

Animals↗
PubMed1988

Methylation of halogenated phenols and thiophenols by cell extracts of gram-positive and gram-negative bacteria.

O-methylation of 2,6-dibromophenol was studied in cell extracts prepared from Rhodococcus sp. strain 1395. O-methylation activity was enhanced by the addition of S-adenosyl-l-methionine but was not affected by the addition of 5-methyltetrahydrofolate nor by up to 10 mM MgCl(2) or EDTA. By using 2,6-dibromophenol, 4,5,6-trichloroguaiacol, and pentachlorothiophenol as the substrates, O-methylation activity was also demonstrated in extracts from two other Rhodococcus sp. strains, an Acinetobacter sp. strain, and a Pseudomonas sp. strain. A diverse range of chloro- and bromophenols, chlorothiophenols, chloro- and bromoguaiacols, and chloro- and bromocatechols were assayed as the substrates by using extracts prepared from strain 1395; all of the compounds were methylated to the corresponding anisoles, veratroles, or guaiacols, which have been identified previously from experiments using whole cells. The specific activity of the enzyme towards the thiophenols was significantly higher than it was towards all the other substrates-high activity was found with pentafluorothiophenol, although the activity with pentafluorophenol was undetectable with the incubation times used. For the chlorophenols, the position of the substituents was of cardinal importance. The enzyme had higher activity towards the halogenated catechols than towards the corresponding guaiacols, and selective O-methylation of the 3,4,5-trihalogenocatechols yielded predominantly the 3,4,5-trihalogenoguaiacols. As in experiments with whole cells, neither 2,4-dinitrophenol, hexachlorophene, nor 5-chloro- or 5-bromovanillin was O-methylated. The results showed conclusively that the methylation reactions were enzymatic and confirmed the conclusion from extensive studies using whole cells that methylation of halogenated phenols may be a significant alternative to biodegradation.

Journal Article↗
PubMed1988

Transformations of halogenated aromatic aldehydes by metabolically stable anaerobic enrichment cultures.

Metabolically stable enrichment cultures of anaerobic bacteria obtained by elective enrichment of sediment samples from the Baltic Sea and Gulf of Bothnia have been used to study the oxidation and reduction of the aldehyde group of various halogenated aromatic aldehydes. During the transformation of 5- and 6-chlorovanillin, 6-bromovanillin, 3-chloro-4-hydroxybenzaldehyde, 3,5-dichloro-4-hydroxybenzaldehyde, and 3,5-dibromo-4-hydroxybenzaldehyde, it was shown that synthesis of the corresponding carboxylic acids, which were the principal metabolites, was invariably accompanied by partial reduction of the aldehyde to a hydroxymethyl group in yields of between 3 and 30%. Complete reduction to a methyl group was observed with some of the halogenated vanillins, but to an extremely limited extent with the halogenated 4-hydroxybenzaldehydes. One consortium produced both the hydroxymethyl and methyl compounds from both 5- and 6-chlorovanillin: it was therefore assumed that the methyl compound was the ultimate reduction product. On the basis of the kinetics of formation of the metabolites, it was concluded that the oxidation and reduction reactions were mechanistically related. In addition to these oxidations and reductions, dehalogenation was observed with one of the consortia. In contrast to the transformations of 5- and 6-chlorovanillin, which produced chlorinated methylcatechols, the corresponding compounds were not observed with 5- and 6-bromovanillin: the former was debrominated, forming 4-methylcatechol, whereas the latter produced 6-bromovanillyl alcohol without demethylation. Similarly, although 3-chloro-4-hydroxybenzaldehyde formed the chlorinated carboxylic acid and the benzyl alcohol, the 3-bromo compound was debrominated with formation of 4-hydroxybenzoic acid and, ultimately, phenol. On prolonged incubation, the halogenated carboxylic acids were generally decarboxylated, so that the final products from these substrates were halogenated catechols or phenols. Reductive processes of the type revealed in this study might therefore plausibly occur in the environment during anaerobic transformation of halogenated aromatic aldehydes containing hydroxyl and/or methoxyl groups.

Journal Article↗
PubMed1987

Transformations of chloroguaiacols, chloroveratroles, and chlorocatechols by stable consortia of anaerobic bacteria.

Metabolically stable consortia of anaerobic bacteria obtained by enrichment of sediment samples with 3,4,5-trimethoxybenzoate (TMBA), 3,4,5-trihydroxybenzoate (gallate [GA]), or 5-chlorovanillin (CV) were used to study the anaerobic transformation of a series of chloroveratroles, chloroguaiacols, and chlorocatechols used as cosubstrates. Experiments were carried out with growing cultures, and the following pathways were demonstrated for metabolism of the growth substrates: (i) TMBA produced GA, which was further degraded without the formation of aromatic intermediates; (ii) GA formed pyrogallol, which was stable to further transformation; and (iii) CV was degraded by a series of steps involving de-O-methylation, oxidation of the aldehyde group, and decarboxylation to 3-chlorocatechol before ring cleavage. Mono-de-O-methylation of the cosubstrates occurred rapidly in the order 4,5,6-trichloroguaiacol greater than 3,4,5-trichloroguaiacol approximately 3,4,5-trichloroveratrole approximately tetrachloroveratrole greater than tetrachloroguaiacol and was concomitant with degradation of the growth substrates. For the polymethoxy compounds--chloroveratroles, 1,2,3-trichloro-4,5,6-trimethoxybenzene, and 4,5,6-trichlorosyringol--de-O-methylation took place sequentially. The resulting chlorocatechols were stable to further transformation until the cultures had exhausted the growth substrates; selective dechlorination then occurred with the formation of 3,5-dichlorocatechol from 3,4,5-trichlorocatechol and of 3,4,6-trichlorocatechol from tetrachlorocatechol. 2,4,5-, 2,4,6-, and 3,4,5-trichoroanisole and 2,3,4,5-tetrachloroanisole were de-O-methylated, but the resulting chlorophenols were resistant to dechlorination. These results extend those of a previous study with spiked sediment samples and their endogenous microflora and illustrate some of the transformations of chloroguaiacols and chlorocatechols which may be expected to occur in anaerobic sediments.

Anisoles↗
PubMed1987

Bacterial O-methylation of halogen-substituted phenols.

Two strains of bacteria capable of carrying out the O-methylation of phenolic compounds, one from the gram-positive genus Rhodococcus and one from the gram-negative genus Acinetobacter, were used to examine the O-methylation of phenols carrying fluoro-, chloro-, and bromo-substituents. Zero-order rates of O-methylation were calculated from data for the chloro- and bromophenols; there was no simple relationship between the rates of reaction and the structure of the substrates, and significant differences were observed in the responses of the two test organisms. For the gram-negative strain, the pattern of substitution was as important as the number of substituents. Hexachlorophene was resistant to O-methylation by both strains, and tetrabromobisphenol-A was O-methylated only by the gram-positive strain. It is suggested that in the natural environment, bacterial O-methylation of phenols carrying electron-attracting substituents might be a significant alternative to biodegradation.

Anisoles↗
PubMed1986

Biotransformations of chloroguaiacols, chlorocatechols, and chloroveratroles in sediments.

The occurrence of trichloro- and tetrachloroguaiacols, -catechols, and -veratroles and their transformation was studied in freshwater and brackish water sediments putatively exposed to bleachery discharge. The samples contained both chloroguaiacols and chlorocatechols, of which >90% could not be removed by simple extraction. The bound concentrations varied and ranged from 550 mug kg of organic C for 3,4,5-trichloroguaiacol to 8,250 mug kg of organic C for tetrachlorocatechol. Chlorinated substrates added to the aqueous phase were rapidly bound to the sediment with K(p) values between 1.3 and 2.8 ml kg of organic C for the chloroguaiacols and chloroveratroles and 22 to 36 ml kg of organic C for the chlorocatechols. Sediment samples incubated aerobically brought about O-methylation of 4,5,6-trichloroguaiacol to 3,4,5-trichloroveratrole in a yield of ca. 25%. Under anaerobic conditions, however, de-O-methylation of both the chloroguaiacols and chloroveratroles took place with synthesis of the corresponding chlorocatechols. In separate experiments, the chlorocatechols were not completely stable under anaerobic conditions, but their ultimate fate has not yet been resolved. Sediment which had been autoclaved twice at 121 degrees C for 20 min was unable to bring about any of these transformations; we therefore conclude that they were mediated by biological processes. These results emphasize that, in determining the fate of chloroguaiacols and related compounds discharged into the aquatic environment, the cardinal roles of sorption to the sediment phase and of the oxygen tension must be taken into account. We propose a hypothetical guaiacol cycle to accommodate our observations.

Journal Article↗
PubMed1985

Short-term test for predicting the potential of xenobiotics to impair reproductive success in fish.

Short-term screening tests with the zebra fish (Brachydanio rerio) have been developed for predicting the potential of xenobiotics to impair reproductive success in fish. The aim was to find simple and sensitive test parameters and to simulate exposure situations typical for anadromous fish species (salmonids), which generally cross heavily polluted coastal areas or estuaries before they reach uncontaminated upstream spawning areas. Therefore, particular attention was directed to tests designed to assess adverse effects induced during gametogenesis in adult fish. the test protocol involves exposure of adults prior to, but not during, spawning and the effects are measured in the offspring as alterations in hatching frequency and hatching rate of eggs, and survival and stress tolerance of embryos and larvae. Some representative examples of the application of these tests are given, and it is shown that impairment of reproductive success can be induced by exposure of parent fish prior to spawning at concentrations of xenobiotics at least five times lower than those yielding effects during direct exposure of embryos and larvae. It is suggested that, in hazard assessment programs, tests of the effect of xenobiotics on the offspring of preexposed adults be routinely incorporated.

Animals↗
PubMed1985

Bacterial o-methylation of chloroguaiacols: effect of substrate concentration, cell density, and growth conditions.

O-methylation of chloroguaiacols has been examined in a number of gram-positive and gram-negative bacteria to elucidate the effects of substrate concentration, growth conditions, and cell density. Substrate concentrations between 0.1 and 20.0 mg liter were used, and it was found that (i) yields of the O-methylated products were significantly higher at the lowest concentrations and (ii) rates of O-methylation were not linear functions of concentration. With 3,4,5-trichloroguaiacol, the nature of the metabolites also changed with concentration. During growth with a range of substrates, O-methylation of chloroguaiacols also took place. With vanillate, however, de-O-methylation occurred: the chlorocatechol formed from 4,5,6-trichloroguaiacol was successively O-methylated to 3,4,5-trichloroguaiacol and 3,4,5-trichloroveratrole, whereas that produced from 4,5-dichloroguaiacol was degraded without O-methylation. Effective O-methylation in nonproliferating suspensions occurred at cell densities as low as 10 cells ml, although both the yields and the rates were lower than in more dense cultures. By using disk assays, it was shown that, compared with their precursors, all of the O-methylated metabolites were virtually nontoxic to the strains examined. It is therefore proposed that O-methylation functions as a detoxification mechanism for cells exposed to chloroguaiacols and chlorophenols. In detail, significant differences were observed in the response of gram-positive and gram-negative cell strains to chloroguaiacols. It is concluded that bacterial O-methylation is to be expected in the natural environment subjected to discharge of chloroguaiacols.

Journal Article↗
PubMed1983

Bacterial methylation of chlorinated phenols and guaiacols: formation of veratroles from guaiacols and high-molecular-weight chlorinated lignin.

Two strains of bacteria, provisionally assigned to the genus Arthrobacter, were shown to metabolize mono-, di-, tri-, and tetrachloroguaiacols and pentachlorophenol to the corresponding O-methyl compounds. Hydroxylated intermediates were formed only transiently, except for the synthesis by one strain of 3,4,5-trichlorosyringol from 3,4,5-trichloroguaiacol. Two isomeric trichloroveratroles and tetrachloroveratrole were formed by three of the strains from a high-molecular-weight chlorinated lignin isolated from kraft pulp mill bleach plant. The concentrations of methylated metabolites varied widely and did not appear to be correlated with degradation. The possible environmental consequences resulting from synthesis of these highly lipophilic substances are discussed briefly.

Journal Article↗
PubMed1976

Acetylene reduction (nitrogen fixation) by enterobacteriaceae isolated from paper mill process waters.

Using selective media containing galactitol, over 130 Enterobacteriaceae have been isolated from paper mill process waters collected from different localities. These bacteria were extensively characterized and tested for acetylene-reducing (nitrogen-fixing) activity under anaerobic conditions. High activity was found in representatives of Klebsiella pneumoniae, Enterobacter aerogenes, Enterobacter cloacae, Erwinia herbicola, Citrobacter freundii, Citrobacter intermedius, and Escherichia coli. Under argon, nitrogenase synthesis was generally not repressed by 5 mM l-glutamate, l-aspartate, l-leucine or Casamino Acids (0.5 g/liter). In many strains, both the specific activities (nanomoles of C(2)H(4) per minute per milligram of protein) and the activities (nanomoles of C(2)H(4) per minute) had considerably declined after 24 h. In three selected strains, activity in intact cells grown under nitrogen was unaffected by the presence during assay of 10 mM l-amino acids or ammonium acetate. All of the strains examined were tolerant towards inactivation of nitrogen-fixing activity by 1.8% (vol/vol) oxygen during assay, and inactivation by up to 10% oxygen was partly reversible. Representatives of the six taxa synthesized nitrogenase in stirred aerobic cultures, though the protein concentrations attained were lower than under anaerobic conditions. It seems reasonable to suggest that under natural conditions, nitrogen fixation is able to contribute significantly to the nitrogen economy of the cells.

Journal Article↗
PubMed1975

Regulation of nitrogenase synthesis in intact cells of Rhodospirillum rubrum: inactivation of nitrogen fixation by ammonia, L-glutamine and L-asparagine.

The synthesis of nitrogenase by intact cells of Rhodospirillum rubrum was repressed in N-free media supplemented with L-glutamine or L-asparagine, but was unaffected by the presence of L-glutamate, L-aspartate or L-histidine. Specific activities attained by cultures in supplemented media maintained under Ar-CO2 were 2 to 3 times higher than those in N-free medium under N2-CO2. A loss in total activity occurred both in cultures growing with N2 after maximum activity had been reached, and in cultures maintained under Ar when the gas phase was changed to N2. There was a rapid loss in nitrogen-fixing activity when low concentrations of NH4+, L-glutamine or L-asparagine were added to cultures with high activities, but this could be recovered in the absence of demonstrable protein synthesis. During growth, the degree of inactivation brought about by 0-5 mM-inactivator increased to 80 to 90%, and NH4+ excreted into the medium reached a maximum concentration towards the end of exponential growth.

Acetylene↗