Biodegradation of oxadiazon by a soil isolated Pseudomonas fluorescens strain CG5: Implementation in an herbicide removal reactor and modelling

An oxadiazon-degrading bacterial, Pseudomonas strain CG5, was isolated from an agricultural contaminated soil. This strain CG5 was able to grow on 10mg of oxadiazon per l, yielding 5.18+/-0.2 mg of protein biomass mol(-1). GC-MS analyses of the metabolites from oxadiazon catabolism revealed its dehalogenation and degradation to form non-toxic end-products, cells were then immobilized by adsorption on a ceramic support to be used as biocatalysts in herbicide removal biofilm-reactor processes. Seventy-two per cent of the oxadiazon was removed, and the maximum specific substrate uptake rate was 10.63+/-0.5 microg h(-1) mg(-1) prot. A new mathematical model was developed to interpret and predict the behaviour of the bacteria and pollutants in a biofilm-reactor system, to consider biofilm structural and morphological properties.     

Biodegradation of low density polyethylene (LDPE) by certain indigenous bacteria and fungi

Low-density polyethylene (LDPE) is an environmental problem because it is discarded randomly, and because in dumpsites, it does not readily degrade. This study reports evidence of successful biodegradation by two fungal species (Aspergillus flavus MCP₅ and Aspergillus flavus MMP₁₀) and eight bacterial species (Acinetobacter sp. MGP₁, Bacillus sp. MGP₁, Pseudomonas sp. MMP₁, Bacillus sp. MMP₅, Staphylococcus sp. MMP₁₀, Bacillus sp. MGP₁, Micrococcus sp. MMP₅ and Bacillus sp. MMP₁₀). These were demonstrated to have used LDPE as both nitrogen and carbon source. LDPE-users were then characterised and identified using standard microbiological procedures. Biodegradation study was done using selected bacterial and fungal isolates, singly and in consortia, to degrade heat-sterilised ground LDPE in media devoid of carbon source, and carbon and nitrogen source. Biodegradation was monitored using gravimetric methods and Fourier transform infrared spectroscopy. This study revealed some microorganisms can use LDPE as both nitrogen and carbon source in the absence of additives.     

Biodegradation and chiral stability of fipronil in aerobic and flooded paddy soils

Microbiological degradation of the racemic mixture, the enantiopure R- and S-fipronil was examined under both aerobic and flooded conditions in three Chinese paddy soils. The degradation kinectics and enantiomer fraction (EF) were determined by means of high-performance liquid chromatography (HPLC) with chiral Chiralcel OD-H column, while desulfinyl, sulfone and sulfide derivatives were monitored by reversed phase HPLC with diode array detection (DAD). The degradation/transformation of enantiomers of fipronil in the three live soils under aerobic and flooded conditions generally complied with the first-order kinetics (R2 > or = 0.94). The calculated t1/2 values of the enantiomers of fipronil ranged between 21 and 34 days for aerobic incubation experiments and between 8 and 19 days under flooded conditions incubation, respectively. The calculated EF values of fipronil during the incubation time were all close to 0.5, indicating that the degradation/transformation of fipronil was almost nonenantioselective. The main metabolites of fipronil formed in the incubation experiments were sulfone and sulfide derivatives by oxidative and reductive processes, respectively. The oxidative pathway seemed more active. Under flooded conditions, S-fipronil was preferentially degraded in the three soil samples used. The main metabolite was determined as fipronil sulfide. In control experiments, almost no removal of enantiomers of fipronil was observed indicating that the degradation of fipronil in the paddy soil used was attributed to microbial mediated processes under both aerobic and flooded conditions. In addition, no enantiomerization of fipronil was observed in the soil samples examined under both aerobic and flooded conditions. These results for major differences in the degradation of the enantiomers as well as the formation of toxic metabolites may have some implications for better environmental and ecological risks assessment for chiral pesticides.     

A short-term study on the interaction of bacteria,fungi and endosulfan in soil microcosm

Endosulfan is one of the few organic chlorine insecticides still in use today in many developing countries. It has medium toxicity for fish and aquatic invertebrates. In this study, we added different concentrations of endosulfan to a series of soil samples collected from Baihua Park in Jinan, Shandong Province, China. Interactions of exogenous endosulfan, bacteria and fungi were analyzed by monitoring the changes in microbe-specific phospholipid fatty acids (PLFA), residual endosulfan and its metabolites which include; endosulfan sulfate, endosulfan lactone and endosulfan diol during a 9 days incubation period. Our results showed that endosulfan reduced fungi biomass by 47% on average after 9 days, while bacteria biomass increased 76% on average. In addition, we found that endosulfan degraded 8.62% in natural soil (NE), 5.51% in strepolin soil (SSE) and 2.47% in sterile soil (SE). Further analysis of the endosulfan metabolites in NE and SSE, revealed that the amount of endosulfan sulfate (ES) significantly increased and that of endosulfan lactone (EL) slightly decreased in both samples after 9 days. However, that of endosulfan diol (ED) increased in NE and decreased in SSE. After collective analysis our data demonstrated that fungi and bacteria responded differently to exogeous endosulfan, in a way that could promote the formation of endosulfan diol during endosulfan degradation.     

苯酚降解菌CN-6的分离及其生长特性

从台湾某纺织厂污泥中分离得到一株能够将苯酚作为唯一碳源和能源生长的细菌。根据其形态和生理生化反应结合16S rDNA测定分析,该菌被鉴定为假单胞杆菌(Pseudomonas sp.)。降解特性研究表明,该菌降解苯酚的最适pH和温度分别为8.5和30℃,在此条件下,浓度高达500mg/L的苯酚能被该菌在17 h内将完全降解。在动力学研究中,由于高浓度苯酚对CN-6菌株的降解过程存在底物抑制现象,故采用Haldane非竞争性底物抑制模型,计算确定了模型参数μmax、KS和KI分别为0.452 h-12、8.617 mg/L7、82.4 mg/L,该动力学方程对实验数据能很好拟合。     

Biodegradation of thiomersal containing effluents by a mercury resistant Pseudomonas putida strain

Thiomersal, a toxic organomercurial with a strong bactericidal effect, is the most widely used preservative in vaccine production. As a result, vaccine production wastewaters are frequently polluted with thiomersal concentrations above the European limit for mercury effluent discharges for which there is, presently, no remediation technology available. This work proposes a biotechnological process for the remediation of vaccine production wastewaters based on the biological degradation of thiomersal to metallic mercury, under aerobic conditions, by a mercury resistant bacterial strain. The kinetics of thiomersal degradation by a pure culture of Pseudomonas putida spi3 was firstly investigated in batch reactors using a thiomersal amended mineral medium. Subsequently, a continuous stirred tank reactor fed with the same medium was operated at a dilution rate of 0.05 h(-1), and the bioreactor performance and robustness was evaluated when exposed to thiomersal shock loads. In a second stage, the bioreactor was fed directly with a real vaccine wastewater contaminated with thiomersal and the culture ability to grow in the wastewater and remediate it was evaluated for dilution rates ranging from 0.022 to 0.1 h(-1).     

Biodegradation of polycyclic aromatic hydrocarbons by Sphingomonas sp. enhanced by water-extractable organic matter from manure compost

The effectiveness of in-situ bioremediation of polycyclic aromatic hydrocarbons (PAHs) may be inhibited by their low aqueous solubility and strong absorption to soil constituents. The aim of this research was to evaluate the effect of water-extractable organic matter (WEOM) from manure compost on the biodegradation of various PAHs. The aqueous solubilities of PAHs including phenanthrene, pyrene and benzo[a]pyrene under different concentrations of WEOM from cow manure compost were initially evaluated. The contribution of WEOM on the degradation of PAHs by Sphingomonas sp. was then investigated. Dissolution results confirmed the ability of WEOM to increase the apparent solubility of the 3PAHs. Time course of biodegradation also revealed its positive contribution to their removal. For example, the degradation of pyrene was 118% higher in the presence of 1000 mg-C L^− 1 WEOM as compared to the mineral salt medium (MSM) alone after 48 h incubation. In addition, degradation was 12% higher with WEOM than with Glucose-Ammonium nitrate despite the more than 6 times higher cell concentration in the latter. WEOM from other manure composts such as chicken and pig were found to have the same effect. Finally, additional tests confirmed that high molecular weight WEOM (> 1000 Da) contributed mainly to solubility and biodegradation enhancements. On the basis of these results, the increase in apparent solubility of PAHs in WEOM solutions may have a significant impact on their biodegradation. It is postulated that the application of WEOM-rich manure composts may be extended in the in-situ bioremediation of PAHs-polluted soil.     

Biodegradation of pharmaceuticals by Rhodococcus rhodochrous and Aspergillus niger by co-metabolism

This work investigated the possible fate of pharmaceuticals in the environment that are known to be resistant to biodegradation. A co-metabolism approach, adding a readily degradable carbon source, was used to study the biodegradation of some pharmaceuticals. The pharmaceuticals selected were all known to be micro pollutants and frequently used by humans. The microorganisms used primarily were Rhodococcus rhodochrous, known to co-metabolize difficult to degrade hydrocarbons and Aspergillus niger. Because of the long periods of time required for the degradation experiments after growth had reached the stationary phase, it was found to be necessary to correct for water loss from the media. Co-metabolism of carbamazepine, sulfamethizole and sulfamethoxazole was observed and as much as 20% of these compounds could be removed. Small amounts of stable metabolites were observed during the degradation of some of these drugs and these were different from the metabolites obtained from abiotic degradation. A metabolite arising from the biodegradation of sulfamethoxazole by R. rhodochrous was identified.     

Isolation and identification of low-density polyethylene (LDPE) biodegrading bacteria from waste landfill in Yazd

Microorganisms used as a biodegradation technique can provide economic benefits and can be compatible with the environment. The aim of this study was to isolate low-density polyethylene (LDPE) degrading bacteria. The waste samples were collected from Yazd landfills. Biochemical and molecular tests based on 16S rDNA sequencing were done to identify the superior isolates. Biodegradation rate was measured using weight loss measurement, spectroscopic analysis with Fourier-transform infrared (FTIR) spectroscopy and an investigation of surface morphology changes using scanning electron microscopy (SEM). Two biodegrading isolates were identified as Pseudomonas aeruginosa strain SKN1 and strain SKN2. The weight loss of LDPE strips was measured at 10.32%. SEM micrographs showed the surface degradation and colony formation on LDPE strips. The FTIR spectrum revealed the structural changes on LDPE strip surfaces. Bio-decomposition was observed to have taken place and the bacterial strains had a special ability to biodegrade LDPE wastes.     

Biodegradation of endocrine disrupting compounds in harbour seawater and sediments

The biodegradation of three endocrine disrupting compounds was examined using samples of seawater and sediment collected from Halifax Harbour, Nova Scotia, Canada, an urbanized harbour impacted by over two centuries of anthropogenic contamination. Flask experiments, where the samples were mixed to form a slurry were used to monitor the aerobic biodegradation of the synthetic plasticizer bisphenol A (BPA), the natural hormone 17β-estradiol (E2), and the pharmaceutical and contraceptive ethinylestradiol (EE2). Degradation rates followed the order E2 > EE2 > BPA with half-lives of up to 1, 5 and 14 days in seawater, respectively. A rapid initial degradation rate for all three compounds with no apparent lag phase indicated the ability of the microbial community to readily catabolise the chemicals. The formation of unidentified non-persistent intermediate metabolites was observed during the E2 degradation experiments. These degradation rates are more rapid and complete than reported in previous studies, indicating the adaptation of native microbial communities to these contaminants.     

Biodegradation of nitrilotriacetic acid (NTA) and ferric-NTA complex by aerobic microbial granules

Development of mixed-culture microbial granules under aerobic conditions in a sequencing batch reactor (SBR), capable of completely degrading a recalcitrant metal chelating agent is reported. In laboratory-scale reactor studies, the microbial granules degraded 2mM of free nitrilotriacetic acid (NTA) and Fe(III)-NTA completely in 14 and 40 h, respectively. Free NTA was degraded at a specific rate of 0.7 mM (gMLSS)(-1)h(-1), while Fe(III)-NTA was degraded at a specific rate of 0.37 mM (gMLSS)(-1)h(-1). Achievement of significant degradation rates of NTA and ferric-NTA in double-distilled water suggests that the microbial metabolism is not constrained by lack of essential elements. Efficient degradation of recalcitrant synthetic chelating agents by aerobic microbial granules suggests their potential application in a variety of situations where heavy metals or radionuclides are to be co-disposed with metal chelating agents.     

Effects of nutrients on biofilm formation and detachment of a Pseudomonas putida strain isolated from a paper machine

The aim of this study was to determine the effect of varying nutrient conditions on biofilm formation of a Pseudomonas putida strain isolated from a paper machine under controlled conditions. Biofilm accumulation, was investigated using a laminar flow cell reactor in a defined mineral medium. Our results indicate that increasing nutrient concentration (from 0.1 to 0.5gl(-1) glucose, C/N=40, C/P=100) or phosphate concentration (from C/P=200 to C/P=100) increased the rate and extent of biofilm accumulation, however, higher nutrient (1gl(-1) glucose, C/N=40, C/P=100) or phosphate (C/P=50) concentration reduced biofilm accumulation rate because of a higher detachment. The rate and extent of biofilm accumulation increased with nitrogen concentration (from C/N=90 to C/N=20). Detachment is a key parameter that influences biofilm accumulation since the early stage (2h) of colonisation and strongly depends on nutrient conditions. In practice, controlling nutrient levels may be interesting to reduce biofilm formation in the paper industry.     

Biodegradation of nodularin and effects of the toxin on bacterial isolates from the Gulf of Gdańsk

Nodularin (NOD), a cyclic pentapeptide produced by the cyanobacterium Nodularia spumigena, is one of the most abundant natural metabolites occurring in the Baltic Sea. The present study investigated the role of this compound in the interactions between cyanobacteria and other bacteria. The toxin inhibited the growth of 15 out of 32 bacterial strains isolated from water and sediments of the Gulf of Gdańsk, southern Baltic Sea. Most of the bacteria sensitive to NOD belonged to the Proteobacteria phylum. Incubation of nodularin in the presence of the bacterial isolates did not reveal any NOD-degrading activity. However, natural microbial communities from sediment removed the toxin within 5-7 days. Analysis by liquid chromatography/hybrid quadrupole-time-of-flight mass spectrometry with turboion spray (QTOF-LC/MS/MS) revealed seven biodegradation products, including five novel ones. The results showed that not only freshwater microorganisms, but also those living in brackish waters, play an important role in cyanotoxin removal.     

Enhanced biodegradation of mixed phenol and sodium salicylate by Pseudomonas putida in membrane contactors

A polypropylene (PP) hollow fiber membrane contactor was used as a reactor to enhance the biodegradation of equimolar phenol and sodium salicylate (SA) by Pseudomonas putida CCRC 14365 at 30 degrees C and pH 7. Experiments were performed at a fixed initial cell density of 0.025 g/L and in the total substrate level range 5.32-63.8 mM. The degradation experiments by free cells were also studied for comparison. With pristine hydrophobic fibers, the degradation of SA was started only after phenol was completely consumed. Substrate inhibitory effect was avoided due to sufficiently low substrate levels in the cell medium; however, the biodegradation was time consuming. With ethanol-wetted fibers, both substrates were completely degraded much faster than the use of pristine fibers. Although the wetted fibers were unable to prevent movement of substrates through the pores, biofilm formed on the outer surfaces of the fibers could enhance the tolerance limit of substrate toxicity. This greatly extended the treatment range to high-level substrate mixtures, as long as the water was nearly neutral and free of concentrated inorganic salts.     

Biodegradation potential of pure and mixed bacterial cultures for removal of 4-nitroaniline from textile dye wastewater

Environmentally toxic aromatic amines including nitroanilines are commonly generated in dye contaminated wastewater in which azo dyes undergo degradation under anaerobic conditions. The aim of this study was to develop a process for biological treatment of 4-nitroaniline. Three bacteria identified as Acinetobacter sp., Citrobacter freundii and Klebsiella oxytoca were isolated from enrichment cultures of activated sludge on 4-nitroaniline, after which the isolates and the mixed culture were studied to determine optimal conditions for biodegradation. HPLC analyses showed the mixed culture was capable of complete removal of 100 μmol/L of 4-nitroaniline within 72 h under aerobic conditions. There was an inverse linear relationship (R² = 0.96) between the rate of degradation (V) and 4-nitraoaniline concentrations [S] over 100-1000 μmol/L. The bacterial culture was also capable of decolorizing structurally different azo dyes (Acid Red-88, Reactive Black-5, Direct Red-81, and Disperse Orange-3) and also degraded nitrobenzene. Our findings show that enrichment cultures from activated sludge can be effective for the removal of dyes and their toxic intermediates, and that treatment may best be accomplished using an anaerobic-aerobic process.     

Behavior and characteristics of dissolved organic matter during column studies of soil aquifer treatment

Soil column experiments were performed to investigate the behavior and characteristics of dissolved organic matter (DOM) during soil aquifer treatment (SAT), and to differentiate among the mechanisms responsible for the changes in the structural and functional properties of DOM during SAT. To determine the biological transformation of DOM, biodegradability tests using a biodegradation-column system were conducted. DOM was fractionated using XAD resins into 5 fractions: hydrophobic acid (HPO-A), hydrophobic neutral (HPO-N), transphilic acid (TPI-A), transphilic neutral (TPI-N) and hydrophilic fraction (HPI). Dissolved organic carbon (DOC) was removed by 70% during SAT, and the sorption and anaerobic biodegradation in SAT led to a DOC reduction of 27.4%. The significant changes in fluorescence properties of DOM were observed during SAT. However, the sorption and anaerobic biodegradation in SAT seemed to have no significant effect on the chemical structure of fluorescing constituents in DOM. The DOM fractions exhibited different changes in Fourier-transform infrared (FT-IR) spectra characteristics during SAT. Biodegradation resulted in the enrichment of aromatic structures and the decreased content of the oxygen-containing functional groups, such as CO and C-O, in DOM. On the other hand, the production of C-O and amide-2 functional groups occurred as a result of the sorption combined with anaerobic biodegradation in SAT.     

Biodegradation of natural and synthetic estrogens by nitrifying activated sludge and ammonia-oxidizing bacterium Nitrosomonas europaea

This report describes the uses of nitrifying activated sludge (NAS) and ammonia-oxidizing bacterium Nitrosomonas europaea to significantly degrade estrone (E1), 17beta-estradiol (E2), estriol (E3), and 17alpha-ethynylestradiol (EE2). Using NAS, the degradation of estrogens obeyed first-order reaction kinetics with degradation rate constants of 0.056 h(-1) for E1, 1.3 h(-1) for E2, 0.030 h(-1) for E3, and 0.035 h(-1) for EE2, indicating that E2 was most easily degraded. Then, we confirmed that E2 was degraded via E1 by NAS. With/without the ammonia oxidation inhibitor, it was observed that ammonia-oxidizing bacteria in conjunction with other microorganisms in NAS degraded estrogens. Using N. europaea, the degradation of estrogens reasonably obeyed zero-order reaction kinetics, and no remarkable difference is present among the four estrogens degradation rates and it was found that E1 was not detected during E2 degradation period. We suggested that E2 was degraded to E1 in NAS could be caused by other heterotrophic bacteria, not by ammonia-oxidizing bacteria.     

Degradation of acetaminophen by Delftia tsuruhatensis and Pseudomonas aeruginosa in a membrane bioreactor

The incidence and fate of pharmaceuticals in the water cycle impose a growing concern for the future reuse of treated water. Because of the recurrent global use of drugs such as Acetaminophen (APAP), an analgesic and antipyretic drug, they are often detected in wastewater treatment plant (WWTP) effluents, receiving surface waters and drinking water resources. In this study, the removal of APAP has been demonstrated in a membrane bioreactor (MBR) fed with APAP as the sole carbon source. After 16 days of operation, at a hydraulic retention time (HRT) of 5 days, more than 99.9% removal was obtained when supplying a synthetic WWTP effluent with 100 μg APAP L⁻¹. Batch experiments indicated no sorption of APAP to the biomass, no influence of the WWTP effluent matrix, and the capability of the microbial consortium to remove APAP at environmentally relevant concentrations (8.3 μg APAP L⁻¹). Incubation with allylthiourea, an ammonia monooxygenase inhibitor, demonstrated that the APAP removal was mainly associated with heterotrophic bacteria and not with the ammonia-oxidizing bacteria. Two APAP degrading strains were isolated from the MBR biomass and identified as Delftia tsuruhatensis and Pseudomonas aeruginosa. During incubation of the isolates, hydroquinone - a potentially toxic transformation product - was temporarily formed but further degraded and/or metabolized. These results suggest that the specific enrichment of a microbial consortium in an MBR operated at a high sludge age might be a promising strategy for post-treatment of WWTP effluents containing pharmaceuticals.     

Detoxification of selenite and mercury by reduction and mutual protection in the assimilation of both elements by Pseudomonas fluorescens

A study on the assimilation and detoxification of selenium and mercury and on the interaction between these two elements was conducted on Pseudomonas fluorescens. P. fluorescens was able to convert separately both elements to their elemental forms, which are less toxic and biologically less available. To study the converting mechanism of selenite to elemental Se, cells were grown in the presence of various selenite concentrations and several parameters such as extracellular protein concentrations, pH, carbohydrate concentrations, isocitrate dehydrogenase (ICDH) and malic enzyme were monitored. Transmission electron microscopy (TEM) and various analytical methods were applied to confirm the interaction between selenium and cell. The former appeared as a red precipitate localized predominantly in the consumed culture medium. P. fluorescens also resisted to the toxic effect of mercury by converting Hg2+ to the volatile and less toxic form Hg0. Mercury reductase was likely responsible for the conversion of Hg2+ to Hg0. More importantly, the interaction between mercury and selenium was also studied. The presence of selenite significantly reduced the accumulation of mercury in P. fluorescens. It was also interesting to note that mercury appeared to behave as a protecting agent against selenium intoxication as the bioaccumulation of Se was also inhibited by this metal. The formation of Se-Hg complexes could explain this mutual protective effect. No precipitate of elemental Se could be detected when Hg was present in the cultures.     

Behaviour of pharmaceutical products and biodegradation intermediates in horizontal subsurface flow constructed wetland. A microcosm experiment

Horizontal subsurface flow constructed wetlands (SSFCWs) are a cost-effective and sustainable alternative to conventional wastewater treatment plants (WWTPs) for sanitation in small communities. SSFCWs are designed to remove suspended solids and organic matter from wastewater but there is little information on the effect of the characteristics of organic matter on the removal efficiency of specific contaminants. In this paper, carbamazepine, ibuprofen and clofibric acid were continuously injected into two SSFCW microcosms fed with synthetic wastewater containing different organic matter sources: dissolved (glucose) and particulate (starch). The response curves of carbamazepine and ibuprofen were compared with that of clofibric acid, which was used as a conservative tracer. The removal efficiencies were found to be independent of the organic matter type (i.e. dissolved or particulate). Carbamazepine was removed inefficiently (5%) by bed sorption, whereas ibuprofen was removed by degradation (51%). In addition, the behaviour of the two main ibuprofen biodegradation intermediates (carboxy and hydroxy derivatives) supported that the main ibuprofen elimination pathway occurs in aerobic conditions.