Novel metabolic pathways of organochlorine pesticides dieldrin and aldrin by the white rot fungi of the genus Phlebia

White rot fungi can degrade a wide spectrum of recalcitrant organic pollutants, including polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated biphenyls (PCBs). In this experiment, 20 white rot fungi, belonging to genus Phlebia, were investigated for their ability to degrade dieldrin. Based on the screening results, we further investigated Phlebia acanthocystis, Phlebia brevispora, and Phlebia aurea to determine their degradation capacity and metabolic products towards dieldrin and aldrin. The three fungi were able to remove over 50% of dieldrin in a low nitrogen medium, after 42 d of incubation. Three hydroxylated products were detected as metabolites of dieldrin, suggesting that in Phlebia strains, hydroxylation reactions might play an important role in the metabolism of dieldrin. In contrast to dieldrin, aldrin exhibited higher levels of degradation activity. Over 90% of aldrin was removed after 28 d of incubation, and several new metabolites of aldrin in microorganisms, including 9-hydroxyaldrin and two carboxylic acid products, were detected in fungal cultures. These results indicate that the methylene moiety of aldrin and dieldrin molecules might be prone to enzymatic attack by white rot fungi. In this study, we describe for the first time a new metabolic pathway of both compounds by fungi of genus Phlebia.     

Removal of aldrin, dieldrin, heptachlor, and heptachlor epoxide using activated carbon and/or Pseudomonas fluorescens free cell cultures

Degradation of aldrin (1,2,3,4,10,10-Hexachloro-1,4,4a,5,8,8a-hexahydro-1,4:5-8-dimethanonaphthalene), heptachlor (1H-1,4,5,6,7,8,8-heptachloro-3a,4,7,7a-tetrahydro-4,7-methano indene), dieldrin (1aalpha,2beta,2aalpha,3beta,6beta,6aalpha,7beta,7aalpha)-3,4,5,6,9,9-Hexachloro-1a,2,2a,3,6,6a,7,7a-octahydro-2,7:3,6-d-methanonaphtha[2,3-b]oxirene, and heptachlor epoxide (1aalpha, 1bbeta,2alpha,5alpha,5alphabeta,6beta,6aalpha-2,3,4,5,6,7,7-Heptachloro-1a,1b,5,5a,6,6a-hexahydro-2,5-methano-2H-inden[1,2-b]-oxirene) was tested using free cultures of Pseudomonas fluorescens under controlled conditions. Pesticide concentrations were monitored by gas chromatography during 120 h. Percentages of degradation and biodegradation rates (BDR) were calculated. Data showed a trend suggesting a relation between chemical structure and degradability. Degradation kinetics for each pesticide tested showed that the highest degradation rates were found in the first 24 h. Kinetics data were adjusted to an empirical equation in order to predict their behavior, and the correlation coefficients obtained were satisfactory. Gas chromatography/mass spectrometry (GC/MS) analysis of the final extracts allowed the identification of chlordene and monodechlorodieldrin, which have been reported as final metabolite produced in the biodegradation of this kind of compounds. Regarding adsorption of pesticides on activated vegetal carbon, we concluded that removal efficiencies between 95.45 and 97.18% can be reached, depending on the pesticide and the carbon dose applied. The values for K from the Freundlich equation were quite similar for the four pesticides (between 1.0001 and 1.04), whereas the n values were quite different for each pesticide in the following order of affinity: dieldrin > aldrin > heptachlor epoxide > heptachlor. Equilibrium times, very important for scaling up the process, were between 43 min and 1 h, for the heptachlor epoxide and the heptachlor, respectively.     

Removal of Aldrin,Dieldrin, Heptachlor,and Heptachlor Epoxide Using Activated Carbon and/or Pseudomonas fluorescens Free Cell Cultures

Degradation of aldrin (1,2,3,4,10,10-Hexachloro-1,4,4a,5,8,8a-hexahydro-1,4:5-8-dimethanonaphthalene), heptachlor (1H-1,4,5,6,7,8,8-heptachloro-3a,4,7,7a-tetrahydro-4,7-methano indene), dieldrin (1aα,2β,2aα,3β,6β,6aα,7β,7aα)-3,4,5,6,9,9-Hexachloro-1a,2,2a,3,6,6a,7,7a-octahydro-2,7:3,6-d-methanonaphtha[2,3-b]oxirene, and heptachlor epoxide (1aα, 1bβ,2α,5α,5αβ,6β,6aα-2,3,4,5,6,7,7-Heptachloro-1a,1b,5,5a,6,6a-hexahydro-2,5-methano-2H-inden[1,2-b]-oxirene) was tested using free cultures of Pseudomonas fluorescens under controlled conditions. Pesticide concentrations were monitored by gas chromatography during 120 h. Percentages of degradation and biodegradation rates (BDR) were calculated. Data showed a trend suggesting a relation between chemical structure and degradability. Degradation kinetics for each pesticide tested showed that the highest degradation rates were found in the first 24 h. Kinetics data were adjusted to an empirical equation in order to predict their behavior, and the correlation coefficients obtained were satisfactory. Gas chromatography/mass spectrometry (GC/MS) analysis of the final extracts allowed the identification of chlordene and monodechlorodieldrin, which have been reported as final metabolite produced in the biodegradation of this kind of compounds. Regarding adsorption of pesticides on activated vegetal carbon, we concluded that removal efficiencies between 95.45 and 97.18% can be reached, depending on the pesticide and the carbon dose applied. The values for K from the Freundlich equation were quite similar for the four pesticides (between 1.0001 and 1.04), whereas the n values were quite different for each pesticide in the following order of affinity: dieldrin > aldrin > heptachlor epoxide > heptachlor. Equilibrium times, very important for scaling up the process, were between 43 min and 1 h, for the heptachlor epoxide and the heptachlor, respectively.     

Determination of oxidative metabolism in Collembolan Proisotoma minuta (Tullberg)

An oxidative metabolism of Collembolan Proisotoma minuta was determined with a model compound of aldrin and dieldrin in this paper. The seven-day LD(50) values for aldrin, dieldrin, and piperonyl butoxide in salt solution were 0.496, 0.367, and 8.346 mg L(-1), respectively. When P. minuta were exposed to aldrin, dieldrin was the sole metabolite. The conversion of aldrin to dieldrin was known to be catalyzed by P450 monooxygenases system. It has been shown that the synergist piperonyl butoxide inhibited the metabolism of aldrin in P. minuta.     

Enrichment and isolation of a mixed bacterial culture for complete mineralization of endosulfan

In the present study, we isolated three novel bacterial species, namely, Staphylococcus sp., Bacillus circulans-I, and Bacillus circulans-II, from contaminated soil collected from the premises of a pesticide manufacturing industry. Batch experiments were conducted using both mixed and pure cultures to assess their potential for the degradation of aqueous endosulfan in aerobic and facultative anaerobic condition. The influence of supplementary carbon (dextrose) source on endosulfan degradation was also examined. After four weeks of incubation, mixed bacterial culture was able to degrade 71.82 +/- 0.2% and 76.04 +/- 0.2% of endosulfan in aerobic and facultative anaerobic conditions, respectively, with an initial endosulfan concentration of 50 mg l(-1). Addition of dextrose to the system amplified the endosulfan degradation efficiency by 13.36 +/- 0.6% in aerobic system and 12.33 +/- 0.6% in facultative anaerobic system. Pure culture studies were carried out to quantify the degradation potential of these individual species. Among the three species, Staphylococcus sp. utilized more beta endosulfan compared to alpha endosulfan in facultative anaerobic system, whereas Bacillus circulans-I and Bacillus circulans-II utilized more alpha endosulfan compared to beta endosulfan in aerobic system. In any of these degradation studies no known intermediate metabolites of endosulfan were observed.     

Degradation of aldrin and endosulfan in rotary drum and windrow composting

Removal efficiencies, kinetics and degradation pathways of aldrin, endosulfan α and endosulfan β in vegetable waste were evaluated during rotary drum and conventional windrow composting. The highest percentage removal of aldrin, endosulfan α and endosulfan β in rotary drum composting was 86.8, 83.3 and 85.3% respectively, whereas in windrow composting, it was 66.6%, 77.7% and 67.2% respectively. The rate constant of degradation of aldrin, endosulfan α and endosulfan β during rotary drum composting ranged from 0.410–0.778, 0.057–0.076 and 0.009–0.061 day^?1 respectively. The pathways of degradation of these pesticides in composting process were proposed. Metabolites dieldrin and 1 hydroxychlorodene formed during composting of aldrin in the vegetable waste indicated the occurrence of epoxidation reaction and oxidation of bridge carbon of aldrin containing the methylene group. Formation of chloroendic acid and chloroendic anhydride during composting of endosulfan containing vegetable waste support the occurrence of endosulfan sulfate and dehydration reaction respectively.     

Concentration of organochlorine pesticides in wine corks

Wine corks were extracted and analyzed for 23 organochlorine pesticides, including alpha-, beta-, and gamma-hexachlorocyclohexanes (HCH), hexachlorobenzene, DDT, DDE and DDD, chlordane, endosulfan, dieldrin, aldrin, and endrin. This was done to investigate the occurrence, concentrations, composition profiles, and possible sources of organochlorine contamination. All groups of compounds were detected in every sample investigated, with the exception of aldrin and endrin. The total concentrations of organochlorine compounds in all samples ranged from 75-120 ng/g lipid, and for most compounds, the concentrations of organochlorines in cork were consistent with published data for other plant tissues. Differences in the relative abundances of the various classes of organochlorine pesticides were substantial and were probably due to differences in the pesticide usage practices of the various regions in which the cork producing trees were grown.     

Using polymer mats to biodegrade atrazine in groundwater: laboratory column experiments

Large-scale column experiments were undertaken to evaluate the potential of in situ polymer mats to deliver oxygen into groundwater to induce biodegradation of the pesticides atrazine, terbutryn and fenamiphos contaminating groundwater in Perth, Western Australia. The polymer mats, composed of woven silicone (dimethylsiloxane) tubes and purged with air, were installed in 2-m-long flow-through soil columns. The polymer mats proved efficient in delivering dissolved oxygen to anaerobic groundwater. Dissolved oxygen concentrations increased from <0.2 mg l(-1) to approximately 4 mg l(-1). Degradation rates of atrazine in oxygenated groundwater were relatively high with a zero-order rate of 240-380 microg l(-1) or a first-order half-life of 0.35 days. Amendment with an additional carbon source showed no significant improvement in biodegradation rates, suggesting that organic carbon was not limiting biodegradation. Atrazine degradation rates estimated in the column experiments were similar to rates determined in laboratory culture experiments, using pure cultures of atrazine-mineralising bacteria. No significant degradation of terbutryn or fenamiphos was observed under the experimental conditions within the time frames of the study. Results from these experiments indicate that remediation of atrazine in a contaminated aquifer may be achievable by delivery of oxygen using an in situ polymer mat system.     

The dechlorination of cyclodiene pesticides by methanogenic granular sludge

Cyclodiene pesticides: aldrin, isodrin, dieldrin and endrin were dechlorinated by methanogenic granular sludge in spiked batch tests. Initial pesticides concentration was 7 or 9 mgl(-1). Two monodechlorinated analogues were formed during the conversions of aldrin and isodrin. Dieldrin was transformed into two monodechlorinated products as well as into aldrin and two monodechlorinated derivatives of aldrin. In respect of endrin three monodechlorinated and three didechlorinated products were found. The dechlorination of endrin was the most rapid, and was almost complete within 28 days. The dechlorinations of aldrin, isodrin and dieldrin were much slower: over a period of 3 months only 59%, 70% and 88% was transformed, respectively. The amounts of released chloride corresponded 0.54 +/- 0.23, 1.05 +/- 0.25, 1.10 +/- 0.12 of theoretical value for suggested reactions, for aldrin, isodrin and dieldrin respectively. For endrin it was much higher. These transformations did not occur in control samples containing autoclaved granules or in control blank samples without sludge. However, in aldrin spiked blanks, a conversion into a six-chlorinated analogue was found.     

The bacterial metabolism of 4,4′-dichlorobiphenyl,and its suppression by alternative carbon sources

The metabolism of 4,4′-dichlorobiphenyl by mixed cultures of bacteria, isolated from activated sludge, was studied in shake cultures and in soil, both in presence and absence of alternative carbon sources. When 4,4′-dichlorobiphenyl was used as sole carbon source, 4-chlorobenzoic acid and 4,4′-dichloro-2,3-biphenyldiol could be isolated from the culture medium. Polar metabolites however, could not be detected in soil and in media in which alternative carbon sources such as glucose, glycerol, peptone, yeast-extract, humic acid or activated sludge were present. No hydroxylated or carboxylic acid derivatives could be isolated when 2,4′, 5-tri-, 2,2′,5,5′,-textra-, 2,2′,3,4,5′-penta-, 2,2′,3,4,5,5′-hexa- and decachlorobiphenyl were used as the sole carbon sources for incubation with bacteria in shake culture.     

Microbial and kinetic characterization of pure and mixed cultures aerobically degrading 4-nitrophenol

The molecular and kinetic characterization of a microorganism able to aerobically degrade 4-nitrophenol (4NP) is presented. The microorganism was isolated from a mixed culture operating in a laboratory-scale sequencing batch reactor with an aerobic anoxic cycle. It was identified as a member of Ralstonia genus within Betaproteobacteria. It is a gram negative coccobacillum (cell length of 2-3 microm) able to aerobically store lipid inclusions when grown aerobically on nitrophenol as the sole carbon source in the range of tested concentrations (80-320 mg l(-1)). Batch kinetic tests were performed with the pure culture, while the kinetics of the mixed biomass was directly investigated in the reactor. For pure cultures exponential growth was observed, with growth rate values in the range of 2-6 d(-1); in experiments with the mixed cultures 4NP concentrations were correlated with growth using the Haldane equation (k(max) = 0.30 mg 4NP mg(-1) VSSh(-1); K(s) = 55 mg 4NPl(-1) and K(I) = 15 mg 4NPl(-1)). Observed pure culture growth rates were higher than those of mixed cultures. This result can be explained by considering that in mixed culture the biomass is evaluated as volatile suspended solids, including both specialized biomass for 4NP removal and denitrifying bacteria.     

Biodegradation of undecylbenzenesulphonate by mixed methane-oxidizing culture

Biodegradation of undecylbenzenesulphonate (C(11)LAS) was performed in shake flasks at 21 degrees C by using a mixed methanotrophic-heterotrophic culture containing type II methanotrophs. Concentrations of C(11)LAS and aromatic intermediates were determined by reversed-phase high-performance liquid chromatography (RP-HPLC). Methane and carbon dioxide concentrations were measured in headspace samples by using gas chromatography. RP-HPLC analyses of aqueous samples show that the culture MM1 expresses the capability of C(11)LAS transformation in the presence or absence of methane. Simultaneous methane oxidation and C(11)LAS degradation, and the inhibition of both transformation processes by acetylene were observed. This suggests the possibility that C(11)LAS transformation is catalyzed by methane monooxygenase (MMO). Comparable affinity of culture MM1 for both methane and C(11)LAS ( [Formula: see text], respectively), and more than four times higher maximum transformation rate for methane than for C(11)LAS ( [Formula: see text] (dry weight) cells day(-1), respectively), were determined. This further supports the involvement of MMO enzyme system in transformation and suggests a pronounced competitive inhibition of C(11)LAS degradation by methane.     

Fungal degradation of metsulfuron-methyl in pure cultures and soil

A fungal strain capable of utilizing metsulfuron-methyl as sole carbon and energy sources was isolated from a metsulfuron-methyl treated soil. The degradation characteristics of metsulfuron-methyl by this fungal strain were investigated in liquid culture and soil. More than 79% of metsulfuron-methyl at concentrations of 0.10 mgl(-1), 1.0 mgl(-1) and 10.0 mgl(-1) in pure culture was degraded by strain MD after incubation for 7 days, whereas only 5.6%, 8.6% and 13.1% of metsulfuron-methyl were degraded at levels of 0.10 mgl(-1), 1.0mgl(-1) and 10.0 mgl(-1) in the controls, respectively. The incorporation of strain MD into soil was found to substantially increase the degradation of metsulfuron-methyl. Degradation was 7.5 and 3.8 times faster in strain MD amended soils than in sterilized and fresh soils. The results show that addition of the isolated strain MD enhances degradation of metsulfuron-methyl in water and soil.     

Biodegradation of polycyclic aromatic hydrocarbons by a mixed culture

We investigated the potential biodegradation of polycyclic aromatic hydrocarbons (PAHs) by an aerobic mixed culture utilizing phenanthrene as its carbon source. Following a 3-5 h post-treatment lag phase, complete degradation of 5 mg/l phenanthrene occurred within 28 h (optimal conditions determined as 30 degrees C and pH 7.0). Phenanthrene degradation was enhanced by the individual addition of yeast extract, acetate, glucose or pyruvate. Results show that the higher the phenanthrene concentration, the slower the degradation rate. While the mixed culture was also capable of efficiently degrading pyrene and acenaphthene, it failed to degrade anthracene and fluorene. In samples containing a mixture of the five PAHs, treatment with the aerobic culture increased degradation rates for fluorene and anthracene and decreased degradation rates for acenaphthene, phenanthrene and pyrene. Finally, it was observed that when nonionic surfactants were present at levels above critical micelle concentrations (CMCs), phenanthrene degradation was completely inhibited by the addition of Brij 30 and Brij 35, and delayed by the addition of Triton X100 and Triton N101.     

Determination of Oxidative Metabolism in Collembolan Proisotoma minuta (Tullberg)

An oxidative metabolism of Collembolan Proisotoma minuta was determined with a model compound of aldrin and dieldrin in this paper. The seven-day LD₅₀ values for aldrin, dieldrin, and piperonyl butoxide in salt solution were 0.496, 0.367, and 8.346 mg L^?1, respectively. When P. minuta were exposed to aldrin, dieldrin was the sole metabolite. The conversion of aldrin to dieldrin was known to be catalyzed by P450 monooxygenases system. It has been shown that the synergist piperonyl butoxide inhibited the metabolism of aldrin in P. minuta.     

Vertical distribution of organochlorine pesticides in humus along Alpine altitudinal profiles in relation to ambiental parameters

In forest soils along vertical profiles located in different parts of the Alps, concentrations of persistent organic pollutants (POPs), namely organochlorine pesticides (OCPs) like dichlorodiphenyltrichloroethanes (DDTs), hexachlorobenzene (HCB), hexachlorocyclohexanes (HCH), heptachlor, aldrin, dieldrin and mirex, were measured. Though local characteristics of the sites are influenced by numerous factors like orographic and meteorological parameters, forest stand characteristics and humus parameters, we ascertained a marked vertical increase of concentrations of some organochlorine compounds in the soil. On the basis of climatological values of each site, we found that the contamination increase with altitude can be ascribed to a certain ‘cold condensation effect’. In addition, the perennial atmospheric deposition of POPs is controlled by precipitation. Other key parameters explaining the accumulation of POPs are the soil organic carbon stocks, the turnover times, the re-volatilisation and degradation processes, which vary with altitude.     

A column test for leaching of organochlorines from soil by amphiphilic nonionic nanopolymers

Amphiphilic nonionic cross-linked nanopolymers (NPs) were synthesized to examine removal of five organochlorines (OCs), namely lindane, heptachlor, aldrin, dieldrin, endrin, and DDT, from a range of Thai agricultural soils. The synthesized NP particles had polarity characteristics similar to those of nonionic surfactant micelles and were largely in the size range of 55-155 nm. This work aimed to determine the optimal conditions for leaching of OC contaminated soil with NPs and also to investigate the role and influence of soil properties on this leaching. An investigation of the concentrations of aqueous dispersions of these particles found that a concentration of 10 g L(-1). was found most effective in leaching the OCs from a column of spiked soil. The optimal contact time that allowed a NP dispersion and spiked soil to reach equilibrium was 48 h. The results indicated influencing factors for OC removal and soil remediation were properties both of the soil and the compounds themselves. Soil organic carbon (SOC) content and soil texture played an important role on the sorption as well as compound hydrophobicity expressed as log K(OW) values. The removal efficiency was found to be in the range of 85.2-92.8 % for all soil samples and in the order of DDT < aldrin < heptachlor < dieldrin < endrin < lindane regardless of soil type. This order is inversely related to the log K(OC) values of these compounds. For OC compounds with a similar molecular structure, removal efficiency was related to molecular weight (MW).     

Biodegradation of 4-nonylphenol in seawater and sediment

Biodegradation of 14C-labelled nonylphenol at the concentration 11 microg litre (-1) in seawater has been estimated by collection and quantification of the formed labelled carbon dioxide. Initially degradation was very slow but when the microorganisms had become adapted, after four weeks at 11 degrees C, the degradation rate increased rapidly and after 58 days about 50% of 14C from NP was found in the CO2 fraction. In the presence of sediment the initial degradation rate was high and did not increase after longer incubation. Lack of oxygen reduced the degradation rate by half in the presence of sediment.     

A ten-year study of chlorinated hydrocarbon insecticide residues in bovine milk in Illinois, 1972 - 1981

Milk supplies in Illinois were monitored for chlorinated hydrocarbon insecticide residues from 1972 through 1981. The percentage of milk samples that were contaminated with aldrin/dieldrin, DDT and its analogs, and BHC/lindane decreased during the 10-year sampling period. The percentage of samples contaminated with aldrin/dieldrin decreased from 98.7% in 1978 to 5.9% in 1981. The amount of aldrin/dieldrin residues found in contaminated samples also decreased from an average of 0.077 ppm (1972-79) to 0.001 ppm in 1980-81. The levels of chlordane and heptachlor/heptachlor epoxide residues in milk did not decrease, but residues of BHC and DDT decreased gradually over the 10-year period. The percentage of samples that had less than 0.1 ppm of all chlorinated hydrocarbon residues combined ranged from 10.6% to 25.7% during 1972 through 1978 and increased to 36.2% in 1979, 71.1% in 1980, and 61.4% in 1981.     

Organochlorine pesticides in the surface waters of Northern Greece

A survey undertaken in Northern Greece has shown that organochlorine pesticides are present in the surface waters. Surface water samples have been collected seasonally from four rivers and five lakes for a period of two years. Solid-phase extraction followed by gas chromatographic techniques with electron capture detection was used for the determination of the compounds. The most commonly encountered organochlorine pesticides in surface waters were the isomers of hexachlorocyclohexane, aldrin, dieldrin and endosulfan sulfate. In some cases the concentrations detected were higher than the qualitative target levels set by the European Union, especially for hexachlorocyclohexane and aldrin. The occurrence of these compounds in Greek surface waters can be attributed to intense agricultural activity as well as to transboundary pollution.