Biodegradation of hydrogen sulphide by inoculated Rhodococcus sp.zw11 in a pilot-scale biotrickling filter

A strain of autotrophic micro-organism, Rhodococcus sp.zw11, was isolated from pharmaceutical wastewater containing hydrogen sulphide (H₂S). The shape, physiological and biochemical characteristics and oxidation capacity of Rhodococcus sp.zw11 were studied, and the effect of inlet concentration and volumetric loading of H₂S on the removal efficiency was evaluated by the biotrickling filter inoculated with Rhodococcus sp.zw11. The results suggested that the optimal temperature of Rhodococcus sp.zw11 (aerobic bacilli, short rod and gram-negative) was from 20°C to 28°C and the optimal pH was from 5.5 to 6.5. The criteria necessary for a scale-up design of the biotrickling filter were established, and pressure drops at the start and end of the experiment were investigated. The optimal inlet loading could be noted as 180 g/m³h, corresponding to H₂S removal efficiency close to 100%. Furthermore, the inoculated biotrickling filter had good ability to resist shock loading, which was a potential industrialisation method to control H₂S emissions.     

Biodegradation of s-triazine herbicide atrazine by Enterobacter cloacae and Burkholderia cepacia sp. from long-term treated sugarcane-cultivated soils in Kenya

In this study soils from sugarcane-cultivated fields were screened for bacterial species capable of atrazine (6-chloro-N²-ethyl-N⁴-isopropyl-1,3,5-triazine-2,4-diamine) degradation due to long exposure of the soils to this herbicide. To enrich for atrazine degraders, Minimal Salt Medium containing atrazine as the sole N source and glucose as the C source was inoculated with soils impacted with this herbicide and incubated. Bacterial growth was monitored by measuring optical density. The degradation of atrazine was followed by measuring residual atrazine in liquid cultures over a given time period by high performance liquid chromatography. Bacterial strains isolated from the enrichment cultures were characterized by biochemical tests and identified by 16S rRNA gene sequencing. Two bacterial strains coded ISL 8 and ISL 15 isolated from two different fields were shown to have 94 and 96% 16S rRNA gene sequence similarity to Burkholderia cepacia respectively. Another bacterial sp., ISL 14 was closely related to Enterobacter cloacae with a 96% 16S rRNA gene sequence similarity. There was not much difference between the extents of atrazine degradation by the enrichment cultures with communities (79–82% applied amount) from which pure strains were isolated and the pure strains themselves in liquid cultures that showed a degradation of 53–83% of applied amount. The study showed existence of bacterial strains in different sugarcane-cultivated fields which can use atrazine as a nitrogen source. The bacterial strains isolated can be used to enhance the degradation of atrazine in contaminated soils where atrazine is still considered to be recalcitrant.     

Studies on Biodegradation of Nicotine by Arthrobacter sp. Strain HF-2

A nicotine-degrading bacterium, strain HF-2, was isolated from tobacco waste-contaminated soil and identified as a member of Arthrobacter sp. based on morphology, physiological tests, 16S rDNA sequence and phylogenetic characteristics. At thermal denaturation test indicated that the G + C mol% of strain HF-1 was 63.5. The relationship between the growth of the isolate and the nicotine degradation suggested that strain HF-2 could utilize nicotine as sole sources of carbon, nitrogen and energy. Blue pigment was observed during the nicotine degradation by strain HF-2. The isolate grew well at 20 to 33°C, initial pH 6.5 to 8.0 and 0.5 to 2.0 g L^?1 of nicotine concentration in the nicotine inorganic salt media. The maximum growth and nicotine degradation occurred at 30°C, initial pH 7.0 and 0.7 g·L^?1 of nicotine concentration in media under natural incubation condition. Strain HF-2 could degrade 100% of nicotine under the optimized incubation conditions for 43 h. The concentrations of nicotine were monitored by high performance liquid chromatography. This study demonstrates Arthrobacter sp. strain HF-2 had a great ability to degrade nicotine, and it may be available for the application to the bioremediation of environments contaminated by tobacco waste.     

Biodegradation of s-triazine herbicide atrazine by Enterobacter cloacae and Burkholderia cepacia sp. from long-term treated sugarcane-cultivated soils in Kenya

In this study soils from sugarcane-cultivated fields were screened for bacterial species capable of atrazine (6-chloro-N2-ethyl-N?-isopropyl-1,3,5-triazine-2,4-diamine) degradation due to long exposure of the soils to this herbicide. To enrich for atrazine degraders, Minimal Salt Medium containing atrazine as the sole N source and glucose as the C source was inoculated with soils impacted with this herbicide and incubated. Bacterial growth was monitored by measuring optical density. The degradation of atrazine was followed by measuring residual atrazine in liquid cultures over a given time period by high performance liquid chromatography. Bacterial strains isolated from the enrichment cultures were characterized by biochemical tests and identified by 16S rRNA gene sequencing. Two bacterial strains coded ISL 8 and ISL 15 isolated from two different fields were shown to have 94 and 96% 16S rRNA gene sequence similarity to Burkholderia cepacia respectively. Another bacterial sp., ISL 14 was closely related to Enterobacter cloacae with a 96% 16S rRNA gene sequence similarity. There was not much difference between the extents of atrazine degradation by the enrichment cultures with communities (79-82% applied amount) from which pure strains were isolated and the pure strains themselves in liquid cultures that showed a degradation of 53-83% of applied amount. The study showed existence of bacterial strains in different sugarcane-cultivated fields which can use atrazine as a nitrogen source. The bacterial strains isolated can be used to enhance the degradation of atrazine in contaminated soils where atrazine is still considered to be recalcitrant.     

Biodegradation of methyl red by Bacillus sp. strain UN2: decolorization capacity,metabolites characterization,and enzyme analysis

Azo dyes are recalcitrant and refractory pollutants that constitute a significant menace to the environment. The present study is focused on exploring the capability of Bacillus sp. strain UN2 for application in methyl red (MR) degradation. Effects of physicochemical parameters (pH of medium, temperature, initial concentration of dye, and composition of the medium) were studied in detail. The suitable pH and temperature range for MR degradation by strain UN2 were respectively 7.0–9.0 and 30–40 °C, and the optimal pH value and temperature were respectively 8.0 and 35 °C. Mg²⁺ and Mn²⁺ (1 mM) were found to significantly accelerate the MR removal rate, while the enhancement by either Fe³⁺ or Fe²⁺ was slight. Under the optimal degradation conditions, strain UN2 exhibited greater than 98 % degradation of the toxic azo dye MR (100 ppm) within 30 min. Analysis of samples from decolorized culture flasks confirmed biodegradation of MR into two prime metabolites: N,N′dimethyl-p-phenyle-nediamine and 2-aminobenzoic acid. A study of the enzymes responsible for the biodegradation of MR, in the control and cells obtained during (10 min) and after (30 min) degradation, showed a significant increase in the activities of azoreductase, laccase, and NADH-DCIP reductase. Furthermore, a phytotoxicity analysis demonstrated that the germination inhibition was almost eliminated for both the plants Triticum aestivum and Sorghum bicolor by MR metabolites at 100 mg/L concentration, yet the germination inhibition of parent dye was significant. Consequently, the high efficiency of MR degradation enables this strain to be a potential candidate for bioremediation of wastewater containing MR.     

Biodegradation of persistent environmental pollutants by Arthrobacter sp.

Persistent environmental pollutants are a growing problem around the world. The effective control of the pollutants is of great significance for human health. Some microbes, especially Arthrobacter, can degrade pollutants into nontoxic substances in various ways. Here, we review the biological properties of Arthrobacter adapting to a variety of environmental stresses, including starvation, hypertonic and hypotonic condition, oxidative stress, heavy metal stress, and low-temperature stress. Furthermore, we categorized the Arthrobacter species that can degrade triazines, organophosphorus, alkaloids, benzene, and its derivatives. Metabolic pathways behind the various biodegradation processes are further discussed. This review will be a helpful reference for comprehensive utilization of Arthrobacter species to tackle environmental pollutants.

     

Alcaligenes sp.YF11菌对杀灭菊酯的降解机理

测定了降解菌Alcaligenessp.YF11对不同浓度杀灭菊酯的降解及其降解途径。在纯培养系统中,Alcaligenessp.YF11对100mg／L的杀灭菊酯的降解符合零级动力学特征,其降解速率为2．1mg／L·h;50mg／L的杀灭菊酯在24h的降解率为87.5％;10mg／L的杀灭菊酯10h的降解率为71．0％。Alcaligenesso．YF11对杀灭菊酯的降解为矿化作用。     

Biodegradation of insecticide carbofuran by Paracoccus sp. YM3

A bacterium (Paracoccus sp. YM3) capable of degrading carbofuran was isolated from carbofuran-contaminated sludge. The strain was shown to metabolize carbofuran (50 mg L(-1)) to carbofuran-7-phenol in minimal salt medium within 6 days in which the pesticide was the only source of carbon. Carbofuran and its main metabolite were analyzed by high performance liquid chromatography (HPLC). The addition of an other carbon source led to accelerated biodegradation. The relevant degrading-enzyme was intracellular and inducible. A tobacco hypersensitivity experiment showed that YM3 could eliminate carbofuran in soils effectively and safely. This is the first report of a Paracoccus sp. that could degrade carbofuran. The present study may provide a basis for biotreatment of wastewaters and bioremediation of carbofuran-contaminated soils.     

Biodegradation of insecticide carbofuran by Paracoccus sp. YM3

A bacterium (Paracoccus sp. YM3) capable of degrading carbofuran was isolated from carbofuran-contaminated sludge. The strain was shown to metabolize carbofuran (50 mg L^?1) to carbofuran-7-phenol in minimal salt medium within 6 days in which the pesticide was the only source of carbon. Carbofuran and its main metabolite were analyzed by high performance liquid chromatography (HPLC). The addition of an other carbon source led to accelerated biodegradation. The relevant degrading-enzyme was intracellular and inducible. A tobacco hypersensitivity experiment showed that YM3 could eliminate carbofuran in soils effectively and safely. This is the first report of a Paracoccus sp. that could degrade carbofuran. The present study may provide a basis for biotreatment of wastewaters and bioremediation of carbofuran-contaminated soils.     

A comparison of mixture toxicity assessment: examining the chronic toxicity of atrazine, permethrin and chlorothalonil in mixtures to Ceriodaphnia cf. dubia

Pesticides predominantly occur in aquatic ecosystems as mixtures of varying complexity, yet relatively few studies have examined the toxicity of pesticide mixtures. Atrazine, chlorothalonil and permethrin are widely used pesticides that have different modes of action. This study examined the chronic toxicities (7-d reproductive impairment) of these pesticides in binary and ternary mixtures to the freshwater cladoceran Ceriodaphnia cf. dubia. The toxicity of the mixtures was compared to that predicted by the independent action (IA) model for mixtures, as this is the most appropriate model for chemicals with different modes of action. Following this they were compared to the toxicity predicted by the concentration addition (CA) model for mixtures. According to the IA model, the toxicity of the chlorothalonil plus atrazine mixture conformed to antagonism, while that of chlorothalonil and permethrin conformed to synergism. The toxicity of the atrazine and permethrin mixture as well as the ternary mixture conformed to IA implying there was either no interaction between the components of these mixtures and/or in the case of the ternary mixture the interactions cancelled each other out to result in IA. The synergistic and antagonistic mixtures deviated from IA by factors greater than 3 and less than 2.5, respectively. When the toxicity of the mixtures was compared to the predictions of the CA model, the binary mixture of chlorothalonil plus atrazine, permethrin plus atrazine and the ternary mixture all conformed to antagonism, while the binary mixture of chlorothalonil plus permethrin conformed to CA. Using the CA model provided estimates of mixture toxicity that did not markedly underestimate the measured toxicity, unlike the IA model, and therefore the CA model is the most suitable to use in ecological risk assessments of these pesticides.     

Biodegradation of crystal violet by a Shewanella sp. NTOU1

A bacterial isolate, strain NTOU1, originally isolated from the cooling system in an oil refinery could decolorize and detoxify crystal violet under anaerobic conditions. The strain was characterized and identified as a member of Shewanella decolorationis based on Gram staining, morphology characters, biochemical tests, the 16S rRNA gene and the gyrase subunit beta gene (gyrB). The optimum pH value and temperature for decolorization of crystal violet by this strain under anaerobic conditions were pH 8-9 and 30-40 degrees C, respectively. Formate (20 mM) was the best electron donor. Addition of ferric citrate did not inhibit decolorization of crystal violet, the addition of thiosulfate, ferric oxide, or manganese oxide slightly decreased decolorization, while addition of nitrite (20 mM) inhibited the decolorization of crystal violet. By supplementing the medium with formate and ferric citrate and cultivating it under optimum pH and temperature, this strain could remove crystal violet, at a concentration of 1500 mg l(-1), at the rate of 298 mg l(-1) h(-1) (during decolorization the OD(600) of the cell culture increased from approximately 0.6 to approximately 1.2). GC/MS analysis of the degradation products of crystal violet detected the presence of N,N'-bis(dimethylamino) benzophenone (Michler's Ketone), [N,N-dimethylaminophenyl] [N-methylaminophenyl] benzophenone, N,N-dimethylaminobenzaldehyde, N,N-dimethylaminophenol, and 4-methylaminophenol. These results suggest that crystal violet was biotransformed into N,N-dimethylaminophenol and Michler's Ketone prior to further degradation of these intermediates. This paper proposes a probable pathway for the degradation of crystal violet by this Shewanella sp. Cytotoxicity and antimicrobial tests showed that the process of decolorization also detoxify crystal violet.     

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.     

Biodegradation of quinoline by gel immobilized Burkholderia sp

Burkholderia sp, a gram-negative, rod-shaped, aerobe, capable of degrading quinoline was immobilized in calcium alginate gel beads and used for degradation of quinoline in aqueous solution in the reactor. The optimal conditions for immobilization of the microorganism, such as alginate concentration, calcium ion concentration, initial cell loading, hardening time and bead size, were determined with a view to improving the quinoline degradation rate. The characteristics of quinoline degradation by immobilized microbial cells were investigated. The repeated use of immobilized cells for quinoline degradation was performed and the results revealed that the bioactivity of immobilized cells was stable over 100 h in the repeated batch cultivation for quinoline degradation.     

Characterization of the first step involved in enzymatic pathway for microcystin-RR biodegraded by Sphingopyxis sp. USTB-05

Enzymes encoded by genes biodegrading microcystins (MCs) can help reveal the function of genes and biodegradation pathway of MCs. Here the first and important gene (USTB-05-A, 1,008 bp) involved in biodegradation of microcystin-RR (MC-RR) was cloned from Sphingopyxis sp. USTB-05 and firstly expressed in Escherichia coli BL21 (DE3) with an expression vector of pGEX4T-1 successfully. The nucleotide sequences of cloned USTB-05-A possessed 92.5% homology to that of mlA reported in Sphingomonas sp. strain ACM-3962. The deduced amino acid sequences containing the cleavage sites of 26th (alanine) and 27th (leucine) showed 83% identical to that of MlrA. The cell-free extract (CE) of recombinant E. coli BL21 (DE3) containing USTB-05-A had high activity for biodegrading MC-RR. Initial MC-RR of 40 mg L⁻¹ was completely biodegraded under total protein of 350 mg L⁻¹ within 0.25 h. A product derived from MC-RR appeared distinctly with the decrease of MC-RR peak on the profile of HPLC. The product (m/z 1056.5) had molecular weight of 18 higher than that of MC-RR (m/z 1038.7). The findings provided the positive evidences that biodegradation of MC-RR began with the breakage of cyclic MC-RR and then it was converted to linear MC-RR as the first product catalyzed by first enzyme of Sphingopyxis sp. USTB-05.     

Biodegradation of acetochlor by a newly isolated Achromobacter sp. strain D-12

A highly effective acetochlor-degrading bacterial strain (D-12) was isolated from the soil of a pesticide factory. The strain was identified as Achromobacter sp. based on its 16S rRNA gene sequence. The strain D-12 optimally degrades acetochlor at a pH of 7.0 and a temperature of 30°C in a mineral salts medium (MSM). Approximately 95% of acetochlor was degraded by the stain treated at a concentration of 10 mg L^?1 after 5 days of incubation. A chiral high performance liquid chromatography (HPLC) system was used to study the enantioselectivity during the process. However, no obvious enantioselective biodegradation was observed. The primary biodegradation acetochlor products were identified by high-performance liquid chromatography-mass spectroscopy (HPLC-MS) and gas chromatography-mass spectrometry (GC-MS). The results indicated that the strain D-12 could be applied in the bioremediation of an acetochlor-polluted environment.     

Biodegradation of triazine herbicide metribuzin by the strain Bacillus sp. N1

By enrichment culturing of soil contaminated with metribuzin, a highly efficient metribuzin degrading bacterium, Bacillus sp. N1, was isolated. This strain grows using metribuzin at 5.0% (v/v) as the sole nitrogen source in a liquid medium. Optimal metribuzin degradation occurred at a temperature of 30ºC and at pH 7.0. With an initial concentration of 20 mg L^?1, the degradation rate was 73.5% in 120 h. If the initial concentrations were higher than 50 mg L^?1, the biodegradation rates decreased as the metribuzin concentrations increased. When the concentration was 100 mg L^?1, the degradation rate was only 45%. Degradation followed the pesticide degradation kinetic equation at initial concentrations between 5 mg L^?1 and 50 mg L^?1. When the metribuzin contaminated soil was mixed with strain N1 (with the concentration of metribuzin being 20 mg L^?1 and the inoculation rate of 10¹¹ g^?1 dry soil), the degradation rate of the metribuzin was 66.4% in 30 days, while the degradation rate of metribuzin was only 19.4% in the control soil without the strain N1. These results indicate that the strain N1 can significantly increase the degradation rate of metribuzin in contaminated soil.     

Biodegradation of 4-aminobenzenesulfonate by Ralstonia sp. PBA and Hydrogenophaga sp. PBC isolated from textile wastewater treatment plant

A co-culture consisting of Hydrogenophaga sp. PBC and Ralstonia sp. PBA, isolated from textile wastewater treatment plant could tolerate up to 100 mM 4-aminobenzenesulfonate (4-ABS) and utilize it as sole carbon, nitrogen and sulfur source under aerobic condition. The biodegradation of 4-ABS resulted in the release of nitrogen and sulfur in the form of ammonium and sulfate respectively. Ninety-eight percent removal of chemical oxygen demand attributed to 20 mM of 4-ABS in cell-free supernatant could be achieved after 118 h. Effective biodegradation of 4-ABS occurred at pH ranging from 6 to 8. During batch culture with 4-ABS as sole carbon and nitrogen source, the ratio of strain PBA to PBC was dynamic and a critical concentration of strain PBA has to be reached in order to enable effective biodegradation of 4-ABS. Haldane inhibition model was used to fit the degradation rate at different initial concentrations and the parameters μ_max, K_s and K_i were determined to be 0.13 h⁻¹, 1.3 mM and 42 mM respectively. HPLC analyses revealed traced accumulation of 4-sulfocatechol and at least four unidentified metabolites during biodegradation. This is the first study to report on the characterization of 4-ABS-degrading bacterial consortium that was isolated from textile wastewater treatment plant.     

Chemotaxis to atrazine and detection of a xenobiotic catabolic plasmid in Arthrobacter sp. DNS10

Introduction

A plasmid named pDNS10 was detected from an atrazine-degrading strain Arthrobacter sp. DNS10 which has been isolated previously in our laboratory.

Materials and methods

In this paper, a special plasmid-detecting method and drop assays experiments were mainly used to achieve research goals.

Results and discussion

pDNS10 exhibited an excellent stability because it also could be detected even when the strain DNS10 has been subcultured under nonselective conditions for eight times. Over a 48-h incubation period, the OD₆₀₀ of samples inoculated with strain DNS10 and strain DNS10-ST (both of them contained pDNS10) were 0.31 ± 0.042 and 0.305 ± 0.034, respectively ,whereas the OD₆₀₀ of samples inoculated strain without pDNS10 (strain DNS10-PE) was only 0.138 ± 0.018. No atrazine was detected in the inoculated strain DNS10 and strain DNS10-ST samples at this period. Contrarily, the atrazine-degrading rate of strain DNS10-PE was only 5.23 ± 0.71%. Furthermore, both the two types of strains containing pDNS10 confirmed the presence of known degrading genes such as trzN, atzB, and atzC. It suggests that pDNS10 is an atrazine catabolic plasmid. In drop assays experiments, the wild-type strain DNS10 cells were chemotactically attracted to atrazine, whereas strain DNS10-PE showed no chemotaxis to atrazine and hydroxyatrazine. There was some relationship between atrazine degradation and the chemotactic response towards atrazine in strain DNS10.

Conclusions

The biochemical characteristics of pDNS10 and the chemotaxis characteristics of strain DNS10 could help us in better understanding of the mechanism of atrazine degradation by strain DNS10.     

Biodegradation and detoxification of organophosphate insecticide, malathion by Fusarium oxysporum f. sp. pisi cutinase

Efficiencies of two lypolytic enzymes (fungal cutinase and yeast esterase) in malathion degradation were investigated. Surprisingly, degradation rate of malathion by fungal cutinase was very high, i.e. almost 60% of initial malathion (500 mg l(-1)) was decomposed within 0.5 h, and nearly 50% of the degraded malathion disappeared within initial 15 min. With the yeast esterase, despite the same concentration, more than 65% of malathion remained even after 2-day treatment. During enzymatic degradation of malathion, two malathion-derived compounds were detected, and time-course changes in composition were also monitored. In the degradation by both fungal cutinase and yeast esterase, two additional organic chemicals were produced from malathion: malathion monoacid (MMA) and malathion diacid (MDA) by ester hydrolysis. Final chemical composition after 2 d was significantly dependent on the enzyme used. Fungal cutinase produced MDA as a major degradation compound. However in the malathion degradation by yeast esterase, an isomer of MMA was produced in abundance in addition to MDA. Toxic effects of malathion and its final degradation products were investigated using various recombinant bioluminescent bacteria. As a result, the degradation products (including MMA) by esterase severely caused membrane damage and inhibition of protein synthesis in bacterial cells, while in the fungal cutinase processes, malathion was significantly degraded to non-toxic MDA after the extended period (2 days).