Piscinibacter caeni sp. nov., isolated from activated sludge.

A yellowish-pigmented bacterial strain, designated as MQ-18T, was isolated from a sample of activated sludge collected from a pharmaceutical factory in Zhejiang, China. The strain was characterized through a polyphasic taxonomy approach. 16S rRNA gene sequence analysis demonstrated that strain MQ-18T showed high similarities to Piscinibacter defluvii SH-1T (99.7 %) and Piscinibacter aquaticus IMCC1728T (98.4 %), thereby suggesting that it belongs to the genus Piscinibacter. The DNA-DNA relatedness values of this strain to strains SH-1T and IMCC1728T were only 35.4 and 33.3 %, respectively. Cells of MQ-18T were Gram-negative, aerobic, motile, rod-shaped and non-spore forming. This strain exhibited growth at 25-37 °C (optimum: 30 °C) in the presence of 0-3.0 % (w/v) NaCl (optimum, 0 % NaCl) and at pH 5.0-8.0 (pH 7.0). The predominant fatty acids were C12 : 0 (5.5 %), C16 : 0 (33.7 %), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c; 38.5 %), and summed feature 4 (anteiso-C17 : 1 B and/or iso C17 : 1 I; 11.6 %). The main quinone type was ubiquinone-8, and the major polyamines were cadaverine and putrescine. The major polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content was 70.1 mol%. On the basis of its phylogenetic, phenotypic and physiological characteristics, strain MQ-18T is considered to represent a novel species of the genus Piscinibacter, for which the name Piscinibacter caeni sp. nov. is proposed. The type strain is MQ-18T (CCTCC AB 2017223T=JCM 32138T).

Comparative investigation on a hexane-degrading strain with different cell surface hydrophobicities mediated by starch and chitosan.

Bioremediation usually exhibits low removal efficiency toward hexane because of poor water solubility, which limits the mass transfer rate between the substrate and microorganism. This work aimed to enhance the hexane degradation rate by increasing cell surface hydrophobicity (CSH) of the degrader, Pseudomonas mendocina NX-1. The CSH of P. mendocina NX-1 was manipulated by treatment with starch and chitosan solution of varied concentrations, reaching a maximum hydrophobicity of 52%. The biodegradation of hexane conformed to the Haldane inhibition model, and the maximum degradation rate (ν max) of the cells with 52% CSH was 0.72 mg (mg cell)-1·h-1 in comparison with 0.47 mg (mg cell)-1·h-1 for cells with 15% CSH. The production of CO2 by high CSH cells was threefold higher than that by cells at 15% CSH within 30 h, and the cumulative rates of O2 consumption were 0.16 and 0.05 mL/h, respectively. High CSH was related to low negative charge carried by the cell surface and probably reduced the repulsive electrostatic interactions between hexane and microorganisms. The FT-IR spectra of cell envelopes demonstrated that the methyl chain was inversely proportional to increasing CSH values, but proteins exhibited a positive effect to CSH enhancement. The ratio of extracellular proteins and polysaccharides increased from 0.87 to 3.78 when the cells were treated with starch and chitosan, indicating their possible roles in increased CSH.

Interaction of tetrahydrofuran and methyl tert-butyl ether in waste gas treatment by a biotrickling filter bioaugmented with Piscinibacter caeni MQ-18 and Pseudomonas oleovorans DT4.

Bioaugmented biotrickling filter (BTF) seeded with Piscinibacter caeni MQ-18, Pseudomonas oleovorans DT4, and activated sludge was established to investigate the treatment performance and biodegradation kinetics of the gaseous mixtures of tetrahydrofuran (THF) and methyl tert-butyl ether (MTBE). Experimental results showed an enhanced startup performance with a startup period of 9 d in bioaugmented BTF (25 d in control BTF seeded with activated sludge). The interaction parameter I2,1 of control (7.462) and bioaugmented BTF (3.267) obtained by the elimination capacity-sum kinetics with interaction parameter (EC-SKIP) model indicated that THF has a stronger inhibition of MTBE biodegradation in the control BTF than in the bioaugmented BTF. Similarly, the self-inhibition EC-SKIP model quantified the positive effects of MTBE on THF biodegradation, as well as the negative effects of THF on MTBE biodegradation and the self-inhibition of MTBE and THF. Metabolic intermediate analysis, real-time quantitative polymerase chain reaction, biofilm-biomass determination, and high-throughput sequencing revealed the possible mechanism of the enhanced treatment performance and biodegradation interactions of MTBE and THF.

Enhanced biodegradation of n-hexane in a two-phase partitioning bioreactor inoculated with Pseudomonas mendocina NX-1 under chitosan stimulation.

Two-phase partitioning bioreactors (TPPBs) have been extensively used for volatile organic compounds (VOCs) removal. To date, most studies have focused on improving the mass transfer of gas phases/non-aqueous phases (NAPs)/aqueous phases, whereas the NAP/biological phases and gas/biological phases transfer has been neglected. Herein, chitosan was introduced into a TPPB to increase cell surface hydrophobicity (CSH) and improve the n-hexane mass transfer. The performance and stability of the TPPB with chitosan for n-hexane biodegradation were investigated, and it was found out that the TPPB with chitosan achieved maximum removal efficiency and elimination capacity of 80.6% and 26.5 g m-3 h-1, thereby reaching much higher values than those obtained without chitosan (61.3% and 15.2 g m-3 h-1). Chitosan not only obvio usly increased cell surface hydrophobicity and cell dry biomass on the surface of silicone oil, but might also allow hydrophobic cells in aqueous phases to directly capture and biodegrade n-hexane, resulting in an obvious improvement of mass transfer from the gas phase to biomass. Stability enhancement was another attractive advantage from chitosan addition. This study might provide a new strategy for the development of TPPB in the hydrophobic VOCs treatment.

Removal of gaseous tetrahydrofuran via a three-phase airlift bioreactor loaded with immobilized cells of GFP-tagged Pseudomonas oleovorans GDT4.

Tetrahydrofuran (THF) is a common highly toxic cyclic aliphatic ether that frequently exists in waste gases. Removal of gaseous THF is a serious issue with important environmental ramifications. A novel three-phase airlift bioreactor (TPAB) loaded with immobilized cells was developed for efficient THF removal from gas streams. An effective THF-degrading transformant, Pseudomonas oleovorans GDT4, which contains the pTn-Mod-OTc-gfp plasmid and was tagged with a green fluorescent protein (GFP), was constructed. Continuous treatment of THF-containing waste gases was succeeded by the GFP-labelled cells immobilized with calcium alginate and activated carbon fiber in the TPAB for 60 days with >90% removal efficiency. The number of fluorescent cells in the beads reached 1.7 × 1011 cells·g-1 of bead on day 10, accounting for 83.3% of the total number of cells. The amount further increased to 3.0 × 1011 cells·g-1 of bead on day 40. However, it decreased to 2.5 × 1011 cells·g-1 of bead with a substantial increase in biomass in the liquid because of cell leakage and hydraulic shock. PCR-DGGE revealed that P. oleovorans was the dominant microorganism throughout the entire operation. The maximum elimination capacity was affected by empty bed residence time (EBRT). The capacity was only 25.9 g m-3·h-1 at EBRT of 80 s, whereas it reached 37.8 g m-3·h-1 at EBRT of 140 s. This work provides an alternative method for full-scale removal of gaseous THF and presents a useful tool for determining the biomass of a specific degrader in immobilized beads.

[Removal of Nitrate Nitrogen by Microbial Photoelectrochemical Cell: PANI/TiO2-NTs as a Photoanode].

The use of microbial photoelectrochemical cells (MPECs) for the removal of contaminants is a cost-effective and environment-friendly method. Based on the preparation of polyaniline/titanium dioxide nanotube array (PANI/TiO2-NTs) composite photoelectrodes, an MPEC system comprising PANI/TiO2-NTs photoanode and biocathode was constructed and the removal performance of nitrate nitrogen (NO3--N) was studied. The experimental results showed that the PANI/TiO2-NT electrode exhibited the best photoelectric performance when the PANI loading time was 80 s. Compared with the TiO2-NTs electrode, the photocurrent density doubled. The light-driven MPEC system could realize autotrophic denitrification without an external voltage. The biodegradation of NO3--N conformed to the pseudo first-order kinetics. The higher the photoresponse current density, the better the denitrification performance of the system. When the initial concentration of NO3--N was 25 mg·L-1 and the photoresponse current density increased from 0.17 mA·cm-2 to 0.67 mA·cm-2, the average denitrification rate increased from 0.83 mg·(L·h)-1 to 2.83 mg·(L·h)-1. High-throughput sequencing of the biocathode microbial membranes revealed that Pseudomonas (27.37%) was the dominant bacteria. It was considered that the photogenerated electrons generated by the PANI/TiO2-NTs photoanode were transmitted to the cathode through an external circuit. Pseudomonas and other microorganisms with autotrophic denitrification and electrochemical activity directly used the electrons on the electrode as the sole electron donors for autotrophic denitrification reaction.

Enhancing Chlorobenzene Biodegradation by Delftia tsuruhatensis Using a Water-Silicone Oil Biphasic System.

In this study, a water-silicone oil biphasic system was developed to enhance the biodegradation of monochlorobenzene (CB) by Delftia tsuruhatensis LW26. Compared to the single phase, the biphasic system with a suitable silicone oil fraction (v/v) of 20% allowed a 2.5-fold increase in the maximum tolerated CB concentration. The CB inhibition on D. tsuruhatensis LW26 was reduced in the presence of silicone oil, and the electron transport system activity was maintained at high levels even under high CB stress. Adhesion of cells to the water-oil interface at the water side was observed using confocal laser scanning microscopy. Nearly 75% of cells accumulated on the interface, implying that another interfacial substrate uptake pathway prevailed besides that initiated by cells in the aqueous phase. The 8-fold increase in cell surface hydrophobicity upon the addition of 20% (v/v) silicone oil showed that silicone oil modified the surface characteristics of D. tsuruhatensis LW26. The protein/polysaccharide ratio of extracellular polymeric substances (EPS) from D. tsuruhatensis LW26 presented a 3-fold enhancement. These results suggested that silicone oil induced the increase in the protein content of EPS and rendered cells hydrophobic. The resulting hydrophobic cells could adhere on the water-oil interface, improving the mass transfer by direct CB uptake from silicone oil.

A solid composite microbial inoculant for the simultaneous removal of volatile organic sulfide compounds: Preparation, characterization, and its bioaugmentation of a biotrickling filter.

Volatile organic sulfide compounds (VOSCs) are usually resistant to biodegradation, thereby limiting the performance of traditional biotechnology dealing with waste gas containing such pollutants especially in mixture. In this study, a solid composite microbial inoculant (SCMI) was prepared to remove dimethyl sulfide (DMS) and propanethiol (PT). Given that the DMS degradation activity of Alcaligenes sp. SY1 is inducible and the PT-degradation activity of Pseudomonas putida S-1 is constitutive, different strategies are designed for cell cultivation to obtain high VOSC removal rates of SCMI. Compared with the microbial suspension, the prepared SCMI exhibited better storage stability at 4 and 25°C. Inoculation of the SCMI in biotrickling filters (BTFs) could effectively shorten the start-up period and enhance the removal performance. Microbial analysis by Illumina MiSeq indicated that Alcaligenes sp. SY1 and P. putida S-1 might be dominant and persistent among the microbial communities of the BTF during the operation.

[Treatment of the Waste Gas Containing Methyl tert-Butyl Ether via a Biotrickling Filter].

The performance and microbial communities of methyl tert-butyl ether (MTBE) treatment using a biotrickling filter (BTF) that was inoculated with activated sewage sludge were investigated. The BTF successfully started up within 23 days when the inlet concentration of MTBE was 100 mg·m-3 and empty bed retention time was 60 s, with 70% removal efficiency (RE). Under steady-state conditions, an elimination capacity (EC) and a mineralization ratio of 13.47 g·(m3·h)-1 and 68% were achieved, respectively. The ECmax was 21.03 g·(m3·h)-1 according to the Haldane model, and a KS of 0.16 g·m-3 and KI of 0.99 g·m-3 were obtained. High-throughput sequencing was used to identify the community structure of the mixed microbial consortium in the BTF. The results indicated that Methylibium sp. (11.33%) and Blastocatella sp. (9.95%) were the dominant bacteria.

[Isolation and Identification of a Chlorobenzene-degrading Bacterium and Its Degradation Characteristics].

A bacterium strain LW26 which could utilize chlorobenzene (CB) as sole carbon and energy source was isolated from a biotrickling filter reactor treating CB-contaminated off-gas. Based on its morphological and physiological characteristics, as well as the analysis of 16S rRNA gene sequence and Biolog test, the strain LW26 was identified as Delftia tsuruhatensis. To our best knowledge, it is the first time that the strain Delftia tsuruhatensis was applied for CB purification. In this study, the effects of temperature, pH, initial CB concentration and Cl- concentration on the biodegradation were investigated. The results showed that the optimal temperature and pH for CB biodegradation were 25℃ and 7.0,respectively; the maximum CB tolerated concentration for LW26 was as high as 500 mg·L-1; when the concentration of Cl- was above 0.14 mol·L-1, the CB degradation was significantly restrained. The degrading process of the strain LW26 followed the Haldane kinetic model and the maximum specific growth rate and the maximum specific degradation rate were 0.42 h-1 and 2.53 h-1, respectively.GC-MS analysis of the metabolites revealed that CB was firstly converted to o-chlorophenol by strain LW26. Combined with the activity of catechol dioxygenase, it can be speculated that CB was finally mineralized to CO2, or converted to cell biomass after processes of ortho cleavage,dechlorination and oxidation.

[Removal of Volatile Sulfur Odor by the Biotrickling Filter].

The biodegradation of gas-phase mixtrue of dimethyl sulfide (DMS) and 1-propanethiol (PT) was examined in a biotrickling filter (BTF), inoculated with a microbial consortium composed of activated sewage sludge, and pure strains of Alcaligenes sp. SY1 and Pseudomonas putida. S-1. BTF could be successfully started up within only 11 days when the inlet concentrations of DMS and PT were both 50 mg·m-3 and EBRT was 30 s, with 90% removal efficiency (RE) of DMS and 100% RE of PT. In the steady state, the maximum elimination capacities of DMS and PT were 8.7 g·(m3·h)-1 and 12.4 g·(m3·h)-1, respectively. The presence of PT with a concentration up to 51 mg·m-3 showed an antagonistic removal pattern for DMS, but the opposite did not occur. Meanwhile, the BTF showed high efficiency in the biodegradation of H2S. When the concentration of H2S was as high as 230 mg·m-3, the RE of H2S could reach 98%. However, H2S showed a declining effect on the removal of DMS when the concentration exceeded 115 mg·m-3.

A newly isolated Pseudomonas putida S-1 strain for batch-mode-propanethiol degradation and continuous treatment of propanethiol-containing waste gas.

Pseudomonas putida S-1 was isolated from activated sludge. This novel strain was capable of degrading malodorous 1-propanethiol (PT). PT degradation commenced with no lag phase by cells pre-grown in nutrition-rich media, such as Luria-Bertani (LB), and PT-contained mineral medium at specific growth rates of 0.10-0.19 h(-1); this phenomenon indicated the operability of a large-scale cell culture. A possible PT degradation pathway was proposed on the basis of the detected metabolites, including dipropyl disulfide, 3-hexanone, 2-hexanone, 3-hexanol, 2-hexanol, S(0), SO4(2-), and CO2. P. putida S-1 could degrade mixed pollutants containing PT, diethyl disulfide, isopropyl alcohol, and acetaldehyde, and LB-pre-cultured cells underwent diauxic growth. Waste gas contaminated with 200-400 mg/m(3) PT was continuously treated by P. putida S-1 pre-cultured in LB medium in a completely stirred tank reactor. The removal efficiencies exceeded 88% when PT stream was mixed with 200 mg/m(3) isopropanol; by contrast, the removal efficiencies decreased to 60% as the empty bed residence time was shortened from 40 s to 20 s.

[Removal of Waste Gas Containing Mixed Chlorinated Hydrocarbons by the Biotrickling Filter].

An experimental investigation on purification of waste gas contaminated with a mixture of dichloromethane (DCM) and dichloroethane(1,2-DCA) was conducted in a biotrickling filter (BTF) inoculated with activated sludge of pharmaceuticals industry. Stable removal efficiency(RE) above 80% for DCM and above 75% for 1,2-DCA were achieved after 35 days, indicating that biofilm was developed. The best elimination capacity (EC) of DCM and 1,2-DCA were 13 g.(m3.h)-1 and 10 g.(m3.h)-1 respectively. And there was a linear relationship between the production of CO2 and mixed gas EC, the maximum mineralization rate of mixed gas stabled at 61. 2%. The interaction test indicated that DCM and 1,2-DCA would inhibit with each other. The changing of biomass of BTF during the operation process was also been studied.

Biodegradation kinetics of tetrahydrofuran, benzene, toluene, and ethylbenzene as multi-substrate by Pseudomonas oleovorans DT4.

The biodegradation kinetics of tetrahydrofuran, benzene (B), toluene (T), and ethylbenzene (E) were systematically investigated individually and as mixtures by a series of aerobic batch degradation experiments initiated by Pseudomonas oleovorans DT4. The Andrews model parameters, e.g., maximum specific growth rates (µmax), half saturation, and substrate inhibition constant, were obtained from single-substrate experiments. The interaction parameters in the sum kinetics model (SKIP) were obtained from the dual substrates. The µmax value of 1.01 for tetrahydrofuran indicated that cell growth using tetrahydrofuran as carbon source was faster than the growth on B (µmax, B = 0.39) or T (µmax, T = 0.39). The interactions in the dual-substrate experiments, including genhancement, inhibition, and co-metabolism, in the mixtures of tetrahydrofuran with B or T or E were identified. The degradation of the four compounds existing simultaneously could be predicted by the combination of SKIP and co-metabolism models. This study is the first to quantify the interactions between tetrahydrofuran and BTE.

[Biodegradation of tetrahydrofuran by combined immobilized of Pseudomonas oleovorans DT4].

A new composite matrix, calcium alginate (CA) coupled with activated carbon fiber (ACF) was designed to immobilize the cells of Pseudomonas oleovorans DT4 for tetrahydrofuran (THF) degradation. The average removal rate of the CA-ACF immobilized cells reached 24.0 mg x (L x h)(-1) with an initial THF concentration of 360 mg x L(-1) when the concentration of CA and ACF was 3% and 1.5% respectively. The mechanical strength of the mobilized cells was also significantly improved with the addition of ACF. Compared to the free suspended cells, higher stable removal efficiency (more than 80%) of CA-ACF cells was detected under different conditions of temperature and pH. The feasibility of the newly designed matrix was also reflected by the repeated batch degradation which showed that the removal activity decreased insignificantly after 80 cycles with the modified reaction system (PNS).

Biodegradation of tetrahydrofuran by Pseudomonas oleovorans DT4 immobilized in calcium alginate beads impregnated with activated carbon fiber: mass transfer effect and continuous treatment.

A novel entrapment matrix, calcium alginate (CA) coupled with activated carbon fiber (ACF), was prepared to immobilize Pseudomonas oleovorans DT4 for degrading tetrahydrofuran (THF). The addition of 1.5% ACF increased the adsorption capacity of the immobilized bead, thus resulting in an enhanced average removal rate of 30.3mg/(Lh). The synergism between adsorption and biodegradation was observed in the hybrid CA-ACF beads instead of in the system comprising CA beads and freely suspended ACF. The effective diffusion coefficient of the CA-ACF bead was not significantly affected by bead size, but the bead's value of 1.14×10(-6)cm(2)/s (for the bead diameter of 0.4 cm) was larger than that of the CA bead by almost one order of magnitude based on the intraparticle diffusion-reaction kinetics analysis. Continuous treatment of the THF-containing wastewater was succeeded by CA-ACF immobilized cells in a packed-bed reactor for 54 d with a >90% removal efficiency.

[Removal of mixed waste gases by the biotrickling filter].

A biotrickling filter (BTF) was designed for treating mixed waste gases, which contained hydrogen sulfide (H2S), tetrahydrofuran (THF) and dichloromethane (DCM) at the start-up and steady states. The removal efficiency of H2S and DCM could maintain about 99% and 60%, respectively, and the removal efficiency of DCM was reduced from 90% to 37% with the shortening empty bed retention time (EBRT) form 50 to 20 seconds when the inlet concentrations were 200, 100, 100 mg x m(-3) of H2S, THF, DCM, respectively. In the theoretical study, the biodegradation efficiency of contaminants was H2S > THF > DCM by analyzing the Michaelis-Menten Dynamic model.

[Influence of nitrate on the simultaneous methanogenesis and denitrification reaction of anaerobic biofilm and granular sludge].

The aims of this study are to further investigate the impact mechanism of nitrate on the simultaneous methanogenesis and denitrification (SMD) process of anaerobic biofilm, and to extend the application of the biofilm process in the treatment of high nitrogen and COD concentration organic wastewater. The SMD reactions were successfully carried out in a hybrid anaerobic biofilm and sludge reactor (HABSR) and an up-flow anaerobic sludge blanket (UASB), and the influence of nitrate on the performance of simultaneous carbon and nitrogen removal in biofilm and granular sludge were investigated using batch tests. The results showed that the nitrate concentration could obviously affect the carbon and nitrogen removal in both biofilm and granular sludge, and the increase of nitrate concentration had more serious impact on the granular sludge, and the biofilm presented higher COD and nitrogen removal efficiency and stronger resistance to toxic materials than the granular sludge. As the nitrate concentration was increased from 75 to 600 mg x L(-1), the COD removal rates were reduced from 273.26 to 0.1 mg x (h x g)(-1) in granular sludge and reduced from 95 to 1.7 mg x (h x g)(-1) in biofilm. At the same time, the denitrification rate of biofilm and granular sludge were increased form 21.43 and 22.31 mg x (h x g)(-1) to 83.72 and 61.06 mg x (h x g)(-1), respectively. The biofilm recovered the COD degradation rate more quickly and easily than the granular sludge, and the maximum COD removal rate reached 712.44 mg x (h x g)(-1). The nitrite accumulation was observed to be the major cause that affected the simultaneous carbon and nitrogen removal of biofilm and granular sludge. It's found that the maximum nitrite accumulation in biofilm was only one tenth of that of the granular sludge at the same nitrate concentration. The HABSR was proved to be an important alternative for SMD reaction employed in the treatment of high nitrogen and COD concentration organic wastewater.