Microbial degradation of 17β -estradiol and 17α -ethinylestradiol followed by a validated HPLC-DAD method

This work aimed at studying the biodegradation of two estrogens, 17α -estradiol (E2) and 17β -ethinylestradiol (EE2), and their potential metabolism to estrone (E1) by microbial consortia. The biodegradation studies were followed by High Performance Liquid Chromatography–Diode Array Detector (HPLC–DAD) using a specifically developed and validated method. Biodegradation studies of the estrogens (E2 and EE2) were carried out with activated sludge (consortium A, CA) obtained from a Wastewater Treatment Plant (WWTP) and with a microbial consortium able to degrade recalcitrant compounds, namely fluorobenzene (consortium B, CB). E2 was more extensively degraded than EE2 by CA whereas CB was only able to degrade E2. The addition of acetate as a supplementary carbon source led to a faster biodegradation of E2 and EE2. E1 was detected as a metabolite only during the degradation of E2. The 16S rRNA gene sequence analyses of strains recovered from the degrading cultures revealed the presence of the genera Pseudomonas, Chryseobacterium and Alcaligenes. The genera Pseudomonas and Chryseobacterium were retrieved from cultures supplied with E2 and EE2, while the genus Alcaligenes was found in the presence of E2, suggesting that they might be involved in the degradation of these compounds.     

Microbial degradation of two carbamate insecticides and their main metabolites in soil

Degradation studies in soil of the insecticides aldicarb and carbofuran and their metabolites (aldicarb sulfoxide, aldicarb sulfone; 3-ketocarbofuran and 3-hydroxycarbofuran) were carried out using laboratory systems under controlled conditions (temperature, water content, light). The insecticides were added to soil samples and subsamples of the soil were analyzed at different times to assess both the bacterial abundance and the concentration of the different chemicals. The epifluorescence direct count method was applied to the subsamples to estimate microorganism numbers (N/g soil). Untreated samples of soil were used as controls for evaluating the effects of the application of the insecticides on microbial abundance. Subsamples treated with the pesticides were analyzed using HPLC and the DT 50 s of the different compounds studied were calculated. The DT 50 values show that neither the parent compounds nor the transformation products have a high persistence in soil and there is a general increase in the concentration of microorganisms as the pesticides diminish.     

Biodegradation and flushing of MBT wastes

Mechanical–biological treatment (MBT) processes are increasingly being adopted as a means of diverting biodegradable municipal waste (BMW) from landfill, for example to comply with the EU Landfill Directive. However, there is considerable uncertainty concerning the residual pollution potential of such wastes. This paper presents the results of laboratory experiments on two different MBT waste residues, carried out to investigate the remaining potential for the generation of greenhouse gases and the flushing of contaminants from these materials when landfilled. The potential for gas generation was found to be between 8% and 20% of that for raw MSW. Pretreatment of the waste reduced the potential for the release of organic carbon, ammoniacal nitrogen, and heavy metal contents into the leachate; and reduced the residual carbon remaining in the waste after final degradation from ～320 g/kg dry matter for raw MSW to between 183 and 195 g/kg dry matter for the MBT wastes.     

Innovative combined technique for high concentration of azo dye AR18 wastewater treatment using modified SBR and enhanced Fenton process as post treatment

The purpose of this research was to evaluate the performance of combined biological/advanced oxidation process (AOP) system for treatment of wastewater containing high concentration (500 mg/L) of azo dye Acid Red 18 (AR18). Two alternating anaerobic–aerobic sequencing batch reactors (SBR1 and SBR2 without and with external feeding at the beginning of aeration cycle, respectively) were operated. The effluent of the SBRs was then post treated through enhanced Fenton process (using zero-valent iron combined with ultrasonic irradiation). More than 90% and 97% of COD was removed in the combined SBR/AOP system without external carbon source (CTS1) and with external feeding (CTS2), respectively. The analysis of dye and its metabolites using UV–vis and HPLC analysis also proved that 99% of the original dye was decolorized and more than 89% of its metabolites were degraded through CST2 which is significantly higher than the reported values in the literature. Besides, more than 87% of phosphorus removal efficiency was obtained in CST2 compared to only 54.5% removal efficiency in CST1. Regarding the findings of this study, the proposed combined treatment system (CTS2) can be suggested as an effective technique for treatment of high azo dye AR18 concentration wastewater.     

Biodegradation of indole by a newly isolated Cupriavidus sp. SHE

Indole, a typical nitrogen heterocyclic aromatic pollutant, is extensively spread in industrial wastewater. Microbial degradation has been proven to be a feasible approach to remove indole, whereas the microbial resources are fairly limited. A bacterial strain designated as SHE was isolated and found to be an efficient indole degrader. It was identified as Cupriavidus sp. according to 16S rRNA gene analysis. Strain SHE could utilize indole as the sole carbon source and almost completely degrade 100 mg/L of indole within 24 hr. It still harbored relatively high indole degradation capacity within pH 4–9 and temperature 25°C–35°C. Experiments also showed that some heavy metals such as Mn^2 +, Pb^2 + and Co^2 + did not pose severe inhibition on indole degradation. Based on high performance liquid chromatography–mass spectrum analysis, isatin was identified as a minor intermediate during the process of indole biodegradation. A major yellow product with m/z 265.0605 (C₁₅H₈N₂O₃) was generated and accumulated, suggesting a novel indole conversion pathway existed. Genome analysis of strain SHE indicated that there existed a rich set of oxidoreductases, which might be the key reason for the efficient degradation of indole. The robust degradation ability of strain SHE makes it a promising candidate for the treatment of indole containing wastewater.     

Biodegradation of pentyl amine and aniline from petrochemical wastewater

The objectives of the project were to isolate a bacterial strain capable of degrading pentyl amine and aniline and to define the optimal pentyl amine and aniline degradation conditions for this bacterial strain. The bacterial strain was isolated from activated sludge obtained from a Northeastern China treatment facility for petrochemical wastewater rich in pentyl amine and aniline. Once the strain was isolated, five triplicate (5) batch tests were used to establish the conditions for maximum pentyl amine and aniline degradation, by varying one at a time the following five factors: temperature, pH, reaction time, pollutant concentrations and aeration rate. In a final test, oil refinery sludge was inoculated with the strain and tested for the degradation of pentyl amine and aniline under optimal conditions, while observing the degradation pathway of pentyl amine and aniline. The isolated strain, PN1001, is a member of the Pseudomonas species and it was capable of degrading pentyl amine and aniline. The optimal reactor conditions for the degradation of a mixture of pentyl amine and aniline, at a concentration ranging between 150 and 200mg/L, were found to be 30 degrees C at a pH of 7.0, under a reaction time of 24h and a maximum solution dissolved oxygen level of 6 mgO(2)/L. Under such conditions, the strain PN1001 degraded 93% and 89% of the pentyl amine and aniline, respectively, aniline being more toxic and demonstrating a more complex degradation pathway. The strain PN1001 degraded more contaminants when both were present because of the pi and sigma electron cloud coordination functions of aniline and pentyl amine, respectively, presumed to reduce the toxic effect of aniline. Once inoculated with the strain, oil refinery sludge degraded 93% and 88% of the pentyl amine and aniline, compared to the strain alone which degraded 72% and 82%, likely because of the sludge's buffering effect against the toxic environment.     

Determination of biodegradation potential by two culture-independent methods in PAH-contaminated soils

Biodegradation potentials of polycyclic aromatic hydrocarbons (PAHs) were determined with soil samples collected from various depths of a PAH-contaminated site and of a site nearby where PAHs were not found. Putative dioxygenase genes were amplified by a primer set specific for initial dioxygenases and identified by web-based database homology search. They were further categorized into several groups of which four dioxygenases were selected as probes for DNA hybridization. The hybridization signals according to the presence of putative dioxygenases were positively related to the extent of PAH contamination. However, the signal intensities varied depending on the probes hybridized and moreover were not consistent with PAH biodegradation activities determined by CO2 evolution. Despite widely accepted advantages of molecular biodegradation assessment, our data clearly present the variations of assessment results depending on the genetic information used and suggest that the methodology may tend to underestimate the real biodegradation capacity of a site probably due to the limited dioxygenase database available at the moment. Therefore, the molecular assessment of biodegradation potential should involve a very careful primer and probe design and an extensive microbiological examination of a site of interest to accurately delineate the biodegradation potential of the site.     

Biodegradation of Methoxychlor and Its Metabolites by the White Rot Fungus Stereum hirsutum Related to the Inactivation of Estrogenic Activity

The white rot fungus Stereum hirsutum was used to degrade methoxychlor [2,2,2-trichloro-1,1-bis(4-methoxyphenyl)ethane] in culture and the degraded products were extensively determined. The estrogenic activity of the degraded products of methoxychlor was examined using cell proliferation and pS2 gene expression assays in MCF-7 cells. S. hirsutum showed high resistance to methoxychlor 100 ppm, and the mycelial growth was fully completed within 8 days of incubation at 30°C. Methoxychlor in liquid culture medium was gradually converted into 2,2-dichloro-1,1-bis(4-methoxyphenyl)ethane, 2,2-dichloro-1,1-bis(4-methoxyphenyl)ethylene, 2-chloro-1,1-bis(4-methoxyphenyl) ethane, 2-chloro-1,1-bis(4-methoxyphenyl) ethylene, and 1,1-bis(4-methoxyphenyl)ethylene, indicating that methoxychlor is dominantly degraded by dechlorination and dehydrogenation. MCF-7 cells were demonstrated to proliferate actively at the 10^?5 M concentration of methoxychlor. However, cell proliferation was significantly inhibited by the incubation with methoxychlor culture media containing S. hirsutum. In addition, the expression level of pS2 mRNA was increased at the concentration (10^?5 M) of methoxychlor. The reductive effect of S. hirsutum for methoxychlor was clear but not significant as in the proliferation assay.     

Determination of the carbon content of biomass—A prerequisite to estimate the complete biodegradation of polymers

This article presents a method to determine the carbon content of biomass, which is formed when degrading biodegradable polymers in an aerobic aqueous test system. Existing methods for determining the carbon content of biomass (e.g., fumigazation, protein assays, dry solids) have several disadvantages when applied for polymer degradation tests. In this work a protein assay based on the Lowry method was used. It was shown that the ratio between protein and carbon content is not constant but depends on the composition of the microbial population, the growth phase, and the substrate supply. This effect was used for the method presented in this article. For determining the carbon content of biomass the absorbance obtained by the Lowry test is correlated directly with the carbon content of biomass in dependence on the duration of the degradation test. The calibration curves are obtained by a mixed population of microorganisms during the course of a degradation test.     

Biodegradation of an Epoxy-Based Thermoplastic Polyester, Poly(Hydroxy Ester Ether) in a Laboratory-Scale Compost System

An epoxy-based thermoplastic polyester, poly(hydroxy ester ether), was incubated under aerobic conditions in a laboratory-scale compost system for 168 days to evaluate its potential for biodegradation. Radiolabeled test polymer [uniformly ¹⁴C ring-labeled, poly(hydroxy ester ether)] was incorporated into a mature compost and a sludge-amended compost at a loading of 3 mg test polymer/g compost. ¹⁴C-Cellulose was used as the positive control and a biologically inhibited control reactor was used to assess abiotic degradation of the test polymer. Degradation of the test polymer was assessed by measuring the amount of ¹⁴C-CO₂ from each of the test reactors. In addition, at selected time intervals subsamples of the compost were collected and serially extracted with water, methanol, and dimethylformamide to monitor degradation of the ¹⁴C-test polymer and provide a partial characterization of the degradation intermediates. Extensive degradation of ¹⁴C-poly(hydroxy ester ether) was observed in the test reactors with degradation half-life of the parent polymer (t _1/2) of approximately 32 days. By the end of the study, only 2% of the total ¹⁴C activity in the test reactors was attributed to intact polymer, with most of the measurable ¹⁴C activity converted to either ¹⁴C-CO₂ (26% of total ¹⁴C activity) or nonextractable products (accounting for 60% of the total activity). In contrast to the test reactors, only 3% of the ¹⁴C-poly(hydroxy ester ether) added to the biologically inhibited control reactor was mineralized to ¹⁴C-CO₂. The results obtained from the microbially active and biologically inhibited compost systems indicate that the poly(hydroxy ester ether) polymer was degraded, at least in part, by a biologically mediated process.