Biofiltration of trimethylamine, dimethylamine, and methylamine by immobilized Paracoccus sp. CP2 and Arthrobacter sp. CP1

A biofilter using granular activated carbon with immobilized Paracoccus sp. CP2 was applied to the elimination of 10–250 ppm of trimethylamine (TMA), dimethylamine (DMA), and methylamine (MA). The results indicated that the system effectively treated MA (>93%), DMA (>90%), and TMA (>85%) under high loading conditions, and the maximum degradation rates were 1.4, 1.2, and 0.9 g-N kg⁻¹ GAC d⁻¹. Among the three different amines treated, TMA was the most difficult to degrade and resulted in ammonia accumulation. Further study on TMA removal showed that the optimal pH was near neutral (6.0–8.0). The supply of high glucose (>0.1%) inhibited TMA removal, maybe due to substrate competition. However, complete TMA degradation was achieved under the co-immobilization of Paracoccus sp. CP2 and Arthrobacter sp. CP1 (96%). Metabolite analysis results demonstrated that the metabolite concentrations decreased by a relatively small 27% while the metabolite apparently increased by heterotrophic nitrification of Arthrobacter sp. CP1 in the co-immobilization biofilter.     

Biodegradation rates of 2-methylisoborneol (MIB) and geosmin through sand filters and in bioreactors

Taste and odour (T&O) causing compounds, in particular, 2-methylisoborneol (MIB) and geosmin, are a problem for water authorities as they are recalcitrant to conventional water treatment. In this study, biological sand filtration was shown to be an effective process for the complete removal of MIB and geosmin, with removal shown to be predominantly through biodegradation. In addition, MIB and geosmin were also effectively degraded in batch bioreactor experiments using biofilm sourced from one of the sand filters as the microbial inoculum. The biodegradation of MIB and geosmin was determined to be a pseudo-first-order reaction with rate constants ranging between 0.10 and 0.58 d⁻¹ in the bioreactor experiments. Rate constants were shown to be dependent upon the initial concentration of the microbial inoculum but not the initial concentration of MIB and geosmin when target concentrations of 200 and 50 ng l⁻¹ were used. Furthermore, rate constants were shown to increase upon re-exposure of the biofilm to both T&O compounds. Enrichment cultures with subsequent community profile analysis using 16S rRNA-directed PCR-DGGE identified four bacteria most likely involved in the biodegradation of geosmin within the sand filters and bioreactors. These included a Pseudomonas sp., Alphaproteobacterium, Sphingomonas sp. and an Acidobacteriaceae member.     

Biodegradation of benzene, toluene; and other aromatic compounds by Pseudmonas sp. D8

Pseudmonas sp. D8 strain, which has the potential to utilize toluene as a sole carbon source, was isolated. At a concentration of 100 mg/l, this strain was found to efficiently degrade toluene and benzene (both individually and in mixture) in culture medium at 30°C and pH7. Following a two-hour lag phase, complete biodegradation of 100 mg/l toluene or benzene occurred within 6 to 8 hours. The addition of nitrate, phosphate, or sulfate at various concentrations were found to have significant influence on both toluene and benzene degradation. In addition, results show that the D8 strain has the ability to degrade monochlorophenols, nitrophenols, and phenol, but not aliphatic compounds. Inoculation of groundwater samples containing 100 mg/1 toluene or benzene with Pseudmonas sp. D8 resulted in rapid degradation within 24 33 hours.     

Combined UV-biological degradation of PAHs

The UV-photolysis of PAHs was tested in silicone oil and tetradecane. In most cases, the degradation of a pollutant provided within a mixture was lower than when provided alone due to competitive effects. With the exception of anthracene, the larger pollutants (4- and 5-rings) were always degraded first, proving that UV-treatment preferentially acts on large PAHs and thereby provides a good complement to microbial degradation. UV-photolysis was also found to be suitable for treatment of soil extract from contaminated soils. The feasibility of UV-biological treatment was demonstrated for the removal of a mixture of phenanthrene and pyrene in silicone oil. UV-irradiation of the silicone oil led to 83% pyrene removal but no phenanthrene photodegradation. Subsequent treatment of the oil in a two-phases partitioning bioreactor (TPPB) system inoculated with Pseudomonas sp. was followed by complete phenanthrene biodegradation but no further pyrene removal. Totally, the combined process allowed 92% removal of the PAH mixture. Further work should focus on characterizing the photoproducts formed and studying the influence of the solvent on the photodegradation process.     

Enhanced biodegradation of endosulfan and its major metabolite endosulfate by a biosurfactant producing bacterium

The present study was carried out to isolate bacteria capable of producing biosurfactant that solublize endosulfan (6,7,8,9,10,10-Hexachloro-1,5,5a,6,9,9a-hexahydro- 6,9-methano-2,4,3-benzodioxathiepine-3-oxide) and for enhanced degradation of endosulfan and its major metabolite endosulfate. The significance of the study is to enhance the bioavailability of soil-bound endosulfan residues as its degradation is limited due to its low solubility. A mixed bacterial culture capable of degrading endosulfan was enriched from pesticide-contaminated soil and was able to degrade about 80% of α-endosulfan and 75% of β-endosulfan in five days. Bacterial isolates were screened for biosurfactant production and endosulfan degradation. Among the isolates screened, four strains produced biosurfactant on endosulfan. ES-47 showed better emulsification of endosulfan and degraded 99% of endosulfan and 94% of endosulfate formed during endosulfan degradation. The strain reduced the surface tension up to 37 dynes/cm. The study reveals that the strain was capable of degrading endosulfan and endosulfate with simultaneous biosurfactant production.     

Extensive biodegradation of polychlorinated biphenyls in Aroclor 1242 and electrical transformer fluid (Askarel) by natural strains of microorganisms indigenous to contaminated African systems

Evidence for substantial aerobic degradation of Aroclor 1242 and Askarel fluid by newly characterized bacterial strains belonging to the Enterobacter, Ralstonia and Pseudomonas genera is presented. The organisms exhibited degradative activity in terms of total PCB/Askarel degradation, degradation of individual congeners and diversity of congeners attacked. Maximal degradation by the various isolates of Askarel ranged from 69% to 86% whereas, Aroclor 1242, with the exception of Ralstonia sp. SA-4 (9.7%), was degraded by 37% to 91%. PCB analysis showed that at least 45 of the representative congeners in Aroclor 1242 were extensively transformed by benzoate-grown cells without the need for biphenyl as an inducer of the upper degradation pathway. In incubations with Aroclor 1242, no clear correlation was observed between percentage of congener transformed and the degree of chlorination, regardless of the presence or absence of biphenyl. Recovery of significant but nonstoichiometric amounts of chloride from the culture media showed partial dechlorination of congeners and suggested production of partial degradation products. Addition of biphenyl evidently enhanced dechlorination of the mixture by some isolates. With the exception of Ralstonia sp. SA-5, chloride released ranged from 24% to 60% in the presence of biphenyl versus 0.35% to 15% without biphenyl.     

Crude oil and hydrocarbon-degrading strains of Rhodococcus rhodochrous isolated from soil and marine environments in Kuwait

Soil and marine samples collected from different localities in Kuwait were screened for microorganisms capable of oil degradation. Both fungi and bacteria were isolated. The fungal flora consisted of Aspergillus terreus, A. sulphureus, Mucor globosus, Fusarium sp. and Penicillum citrinum. Mucor globosus was the most active oil degrading fungus isolated. Bacterial isolates included Bacillus spp. Enterobacteriaceae, Pseudomonas spp., Nocardia spp., Streptomyces spp.,and Rhodococcus spp. Among these Rhodococcus strains were the most efficient in oil degradation and, relatively speaking, the most abundant. Bacterial and fungal isolates differed in their ability to degrade crude oil, with Rhodococcus isolates being more active that fungin in n-alkane biodegradation, particularly in the case of R. rhodochrous. In addition to medium chain n-alkanes, fungi utilized one or more of the aromatic hydrocarbons studied, while bacteria failed to do so. R. rhodochorous KUCC 8801 was shown by GLC and post-growth studies to be more efficient in oil degradation than isolates known to be active oil degraders.     

Biodegradation of nonylphenol in sewage sludge

We investigated the effects of various factors on the aerobic degradation of nonylphenol (NP) in sewage sludge. NP (5 mg/kg) degradation rate constants (k₁) calculated were 0.148 and 0.224 day⁻¹ for the batch experiment and the bioreactor experiment, respectively, and half-lives (t_1/2) were 4.7 and 3.1 days, respectively. The optimal pH value for NP degradation in sludge was 7.0 and the degradation rate was enhanced when the temperature was increased and when yeast extract (5 mg/l) and surfactants such as brij 30 or brij 35 (55 or 91 μM) were added. The addition of aluminum sulfate (200 mg/l) and hydrogen peroxide (1 mg/l) inhibited NP degradation within 28 days of incubation. Of the microorganism strains isolated from the sludge samples, we found that strain CT7 (identified as Bacillus sphaericus) manifested the best degrading ability.     

Characterization of cyclohexane and hexane degradation by Rhodococcus sp. EC1

Cyclohexane is a recalcitrant compound that is more difficult to degrade than even n-alkanes or monoaromatic hydrocarbons. In this study, a cyclohexane-degrading consortium was obtained from oil-contaminated soil by an enrichment culture method. Based on a 16S rDNA polymerase chain reaction-denaturing gradient gel electrophoresis method, this consortium was identified as comprising Alpha-proteobacteria, Actinobacteria, and Gamma-proteobacteria. One of these organisms, Rhodococcus sp. EC1, was isolated and shown to have excellent cyclohexane-degrading ability. The maximum specific cyclohexane degradation rate (Vmax) for EC1 was 246 micromol g-DCW(-1) (dry cell weight)h(-1). The optimum conditions of cyclohexane degradation were 25-35 degrees C and pH 6-8. In addition to its cyclohexane degradation abilities, EC1 was also able to strongly degrade hexane, with a maximum specific hexane degradation rate of 361 micromol g-DCW(-1)h(-1). Experiments using 14C-hexane revealed that EC1 mineralized 40% of hexane into CO2 and converted 53% into biomass. Moreover, EC1 could use other hydrocarbons, including methanol, ethanol, acetone, methyl tert-butyl ether, pyrene, diesel, lubricant oil, benzene, toluene, ethylbenzene, m-xylene, p-xylene and o-xylene. These findings collectively suggest that EC1 may be a useful biological resource for removal of cyclohexane, hexane, and other recalcitrant hydrocarbons.     

Degradation of atrazine in mineral salts medium and soil using enrichment culture

Atrazine degrading enrichment culture was prepared by its repeated addition to an alluvial soil and its ability to degrade atrazine in mineral salts medium and soil was studied. Enrichment culture utilized atrazine as a sole source of carbon and nitrogen in mineral salts medium and degradation slowed down when sucrose and/or ammonium hydrogen phosphate were supplemented as additional source of carbon and nitrogen, respectively. Biuret was detected as the only metabolite of atrazine while deethylatrazine, deisopropyatrazine, hydroxyatrazine and cyanuric acid were never detected at any stage of degradation. Enrichment culture degraded atrazine in an alkaline alluvial soil while no degradation was observed in the acidic laterite soil. Enrichment culture was able to withstand high concentrations of atrazine (110 μg/g) in the alluvial soil as atrazine was completely degraded. Developed mixed culture has the ability to degrade atrazine and has potential application in decontamination of contaminated water and soil.     

Isolation and characterization of an Alcaligenes sp. strain DG-5 capable of degrading DDTs under aerobic conditions

In this study, an Alcaligenes sp. strain DG-5 that can effectively degrade dichlorodiphenyltrichloro-ethanes (DDTs) under aerobic conditions was isolated from DDTs-contaminated sediment. Various factors that affect the biodegradation of DDTs by DG-5 were investigated. About 88 %, 65 % and 45 % of the total DDTs were consumed within 120 h when their initial concentrations were 0.5, 5 and 15 mg L?1, respectively. However, almost no degradation was observed when their concentration was increased to 30 mg L?1, but the addition of nutrients significantly improved the degradation, and 66 % and 90 % of the total DDTs were degraded at 336 h in the presence of 5 g L?1 peptone and yeast extract, respectively. Moreover, the addition of 20 mM formate also enhanced the ability of DG-5 to transform DDTs, and its DDT transformation capacity (T(c)) value was increased by 1.8 - 2.7 fold for the pure (p,p'-DDT or o,p'-DDT only) and mixed systems (p,p'-DDT, o,p'-DDT, p,p'-DDD and p,p'-DDE). Furthermore, it was found that competitive inhibition in the biodegradation by DDT compounds occurred in the mixed system.     

Characterization of hydrocarbon-degrading and biosurfactant-producing Pseudomonas sp. P-1 strain as a potential tool for bioremediation of petroleum-contaminated soil

The Pseudomonas sp. P-1 strain, isolated from heavily petroleum hydrocarbon-contaminated soil, was investigated for its capability to degrade hydrocarbons and produce a biosurfactant. The strain degraded crude oil, fractions A5 and P3 of crude oil, and hexadecane (27, 39, 27 and 13 % of hydrocarbons added to culture medium were degraded, respectively) but had no ability to degrade phenanthrene. Additionally, the presence of gene-encoding enzymes responsible for the degradation of alkanes and naphthalene in the genome of the P-1 strain was reported. Positive results of blood agar and methylene blue agar tests, as well as the presence of gene rhl, involved in the biosynthesis of rhamnolipid, confirmed the ability of P-1 for synthesis of glycolipid biosurfactant. ¹H and ¹³C nuclear magnetic resonance, Fourier transform infrared spectrum and mass spectrum analyses indicated that the extracted biosurfactant was affiliated with rhamnolipid. The results of this study indicate that the P-1 and/or biosurfactant produced by this strain have the potential to be used in bioremediation of hydrocarbon-contaminated soils.     

Biodegradation of 2,4,6-trichlorophenol in the presence of primary substrate by immobilized pure culture bacteria

In this study, pure strains that are capable of utilizing 2,4,6-trichlorophenol have been isolated from the mixed culture grown on substrates containing chlorophenolic compounds. Studies have been carried out on the capability of these isolated pure strains in suspended and immobilized forms to decompose 2,4,6-trichlorophenol. Additionally, the influence of primary substrates (e.g., phenol, 2-chlorophenol, 3-chlorophenol, 4-chlorophenol, 2,4-dichlorophenol) on the decomposition of 2,4,6-trichlorophenol by the isolated pure strains grown in immobilized form is also investigated. The results are: Through bacterial isolation and identification, three pure strains have been obtained: Pseudomonas spp. strain 01, Pseudomonas spp. strain 02 and Agrobacterium spp. Whether in suspended or immobilized forms, all strains have poor removal efficiencies of 2,4,6-trichlorophenol. However, addition of 200 mg/l phenol will enable the immobilized Pseudomonas spp. strain 01, and Pseudomonas spp. strain 02 to achieve 65% and 48% removal of 2,4,6-trichlorophenol, respectively. Addition of phenol will assist the immobilized Pseudomonas spp. strain 02 in achieving removal of 2,4,6-trichlorophenol but the removal efficiency is not good if the phenol concentration is too low. The optimum phenol concentration should be between 200 and 400 mg/l.     

Optimization of methyl parathion biodegradation and detoxification by cells in suspension or immobilized on tezontle expressing the opd gene

The goal of this study was to optimize methyl parathion (O,O-dimethyl-O-4-p-nitrophenyl phosphorothioate) degradation using a strain of Escherichia coli DH5α expressing the opd gene. Our results indicate that this strain had lower enzymatic activity compared to the Flavobacterium sp. ATCC 27551 strain from which the opd gene was derived. Both strains were assessed for their ability to degrade methyl parathion (MP) in a mineral salt medium with or without the addition of glucose either as suspended cells or immobilized on tezontle, a volcanic rock. MP was degraded by both strains with similar efficiencies, but immobilized cells degraded MP more efficiently than cells in suspension. However, the viability of E. coli cells was much higher than that of the Flavobacterium sp. We confirmed the decrease in toxicity from the treated effluents through acetylcholinesterase activity tests, indicating the potential of this method for the treatment of solutions containing MP.     

Low-density polyethylene degradation by Pseudomonas sp. AKS2 biofilm

Polyethylene materials are a serious environmental concern as their nondegradable nature allows them to persist in the environment. Recent studies have shown that polyethylene can be degraded by microbes at a very slow rate, whereby detectable changes are evident after several years. In the present study, we report the degradation of low-density polyethylene by Pseudomonas sp. AKS2. Unlike the previous reports, degradation by Pseudomonas sp. AKS2 is relatively fast as it can degrade 5?±?1 % of the starting material in 45 days without prior oxidation. This degradation can be altered by agents that modulate hydrophobic interaction between polythene and the microbe. As mineral oil promotes hydrophobic interactions, it enhances bacterial attachment to the polymer surface. This enhanced attachment results in increased biofilm formation and enhanced polymer degradation. In contrast, Tween 80 reduces bacterial attachment to the polymer surface by lowering hydrophobic interactions and thereby reduces polymer degradation. Thus, this study establishes a correlation between hydrophobic interaction and polymer degradation and also relates the biofilm formation ability of bacteria to polymer degrading potential.     

Isolation and characterization of an Alcaligenes sp. strain DG-5 capable of degrading DDTs under aerobic conditions

In this study, an Alcaligenes sp. strain DG-5 that can effectively degrade dichlorodiphenyltrichloro-ethanes (DDTs) under aerobic conditions was isolated from DDTs-contaminated sediment. Various factors that affect the biodegradation of DDTs by DG-5 were investigated. About 88 %, 65 % and 45 % of the total DDTs were consumed within 120 h when their initial concentrations were 0.5, 5 and 15 mg L^?1, respectively. However, almost no degradation was observed when their concentration was increased to 30 mg L^?1, but the addition of nutrients significantly improved the degradation, and 66 % and 90 % of the total DDTs were degraded at 336 h in the presence of 5 g L^?1 peptone and yeast extract, respectively. Moreover, the addition of 20 mM formate also enhanced the ability of DG-5 to transform DDTs, and its DDT transformation capacity (T_c) value was increased by 1.8 - 2.7 fold for the pure (p,p’-DDT or o,p’-DDT only) and mixed systems (p,p’-DDT, o,p’-DDT, p,p’-DDD and p,p’-DDE). Furthermore, it was found that competitive inhibition in the biodegradation by DDT compounds occurred in the mixed system.     

Aerobic degradation of diuron by aquatic microorganisms

Abstract

Degradation of diuron [3‐(3,4‐dichlorophenyl)‐l,1‐dimethyl‐urea] by microorganisms obtained from pond water and sediment was determined under aerobic conditions. Enrichment procedures were used to isolate cultures capable of degrading the herbicide. Several mixed fungal/bacterial and mixed bacterial cultures were isolated that could degrade diuron. The mixed cultures degraded 67–99% of the added diuron forming from six to seven products which were separated via TLC. The major degradation product detected in most culture extracts was 3,4‐dichloroaniline. Other identified products formed were 3‐(3,4‐dichlorophenyl)‐1‐methylurea and 3‐(3,4‐dichlorophenyl)urea.     

Influence of microbial biomass on the biodegradability of organic compounds

The influence of different types of inocula as well as the amount of inoculum (microbial biomass) on the biodegradation pattern of acetate, 4-nitrophenol, and the three methoxyaniline isomers was investigated in the Modified OECD test and a NPR guidline. Using sediment of the river Rhine as inoculum 4-nitrophenol could not be degraded, while an inoculum from garden soil gave only 60% degradation in the OECD test. Effluent of an activated sludge plant however was able to degrade 4-nitrophenol at a concentration of 19 mg/1 in the OECD test completely. At a concentration of 94 mg/1 no degradation was observed. Testing the methoxyanilines for biodegradability it was found that at a low inoculum level (OECD protocol) no degradation of the three compounds occurred. Using activated sludge (1.5 ml/1) as inoculum 3- and 4-methoxy aniline could be degraded for 60% respectively completely while 2-methoxyaniline was still refactory to degradation. Measuring the microbial biomass by means of ATP during biodegradation strongly suggested that the microbial flora which rapidly metabolize acetate is quite another microflora than the microflora responsible for the degradation of 4-nitrophenol.     

Removal of a cationic dye from aqueous solutions by adsorption onto bentonite clay

The ability of bentonite to remove malachite green from aqueous solutions has been studied for different adsorbate concentrations by varying the amount of adsorbent, temperature, pH and shaking time. Maximum adsorption of the dye, i.e. >90% has been achieved in aqueous solutions using 0.05 g of bentonite at a pH of 9. Thermodynamic parameters such as ΔH°, ΔS° and ΔG° were calculated from the slope and intercept of the linear plots of ln K_D against 1/T. Analysis of adsorption results obtained at 298, 308, 318 and 328 K showed that the adsorption pattern on bentonite seems to follow the Langmuir, Freundlih and D–R isotherms. The temperature increase reduces adsorption capacity by bentonite, due to the enhancement of the desorption step in the mechanism. The numerical values of sorption free energy (E_a) of 1.00–1.12 kJ mol⁻¹ indicated physical adsorption. The kinetic data indicated an intraparticle diffusion process with sorption being first order. The rate constant k was 0.526 min⁻¹. The concentration of malachite green oxalate was measured before and after adsorption by using UV–Vis spectrophotometer.