Decolorization of textile dyestuffs by a mixed bacterial consortium

A mixed anaerobic bacterial culture decolorized Drimaren Orange K-GL, Everzol Red RBN and Everdirect Supra Yellow PG dyestuffs at 200 mg dyestuff l^–1 over 24 h. Improved performance with complete decolorization within 24 h was achieved by incubation with 5 g yeast extract l^–1 compared to glucose, lactose and sucrose though 50 mg yeast extract l^–1 supplemented with 5 g lactose l^–1 or 5 g sucrose l^–1 also resulted in complete decolorization within 24 h.     

Biodegradation of fuel oil hydrocarbons by a mixed bacterial consortium in sandy and loamy soils

The aerobic degradation of light fuel oil in sandy and loamy soils by an environmental bacterial consortium was investigated. Soils were spiked with 1 or 0.1% of oil per dry weight of soil. Acetone extracts of dried soils were analyzed by GC and the overall degradation was calculated by comparison with hydrocarbon recovery from uninoculated soils. In sandy soils, the sum of alkanes n-C(12) to n-C(23) was degraded to about 45% within 6 days at 20 degrees C and to 27-31% within 28 days, provided that moisture and nutrients were replenished. Degradation in loamy soil was about 12% lower. The distribution of recovered alkanes suggested a preferential degradation of shorter chain molecules (n-C(12) to n-C(16)) by the bacterial consortium. Partial 16S rDNA sequences indicated the presence of strains of Pseudomonas aeruginosa, Pseudomonas citronellolis, and Stenotrophomonas maltophilia. Toxicity tests using commercial standard procedures showed a moderate inhibition of bacterial activity. The study showed the applicability of a natural microbial community for the degradation of oil spills into soils at ambient temperatures.     

Degradation of crude oil by an arctic microbial consortium

The ability of a psychrotolerant microbial consortium to degrade crude oil at low temperatures was investigated. The enriched arctic microbial community was also tested for its ability to utilize various hydrocarbons, such as long-chain alkanes (n-C₂₄ to n-C₃₄), pristane, (methyl-)naphthalenes, and xylenes, as sole carbon and energy sources. Except for o-xylene and methylnaphthalenes, all tested compounds were metabolized under conditions that are typical for contaminated marine liquid sites, namely at pH 6–9 and at 4–27°C. By applying molecular biological techniques (16S rDNA sequencing, DGGE) nine strains could be identified in the consortium. Five of these strains could be isolated in pure cultures. The involved strains were closely related to the following genera: Pseudoalteromonas (two species), Pseudomonas (two species), Shewanella (two species), Marinobacter (one species), Psychrobacter (one species), and Agreia (one species). Interestingly, the five isolated strains in different combinations were unable to degrade crude oil or its components significantly, indicating the importance of the four unculturable microorganisms in the degradation of single or of complex mixtures of hydrocarbons. The obtained mixed culture showed obvious advantages including stability of the consortium, wide range adaptability for crude oil degradation, and strong degradation ability of crude oil.     

Cellulose degradation by a mixed bacterial culture

Summary An integrated mixed bacterial culture consisting of four strains has been isolated by a batch enrichment technique. The cellulolytic member (strain D) is aCellulomonas sp. and the others are non-cellulolytic. The interaction between strains D and C is pronounced and appears to involve an exchange of reducing sugars and growth factors. The symbiotic relationship of this naturally occurring mixed culture is therefore one of mutualism. The filter paper cellulase and carboxymethyl cellulase activities in extracellular fluid are high, while -glucosidase activity is low. The mixed culture digests a variety of lignocellulosics efficiently and is of fundamental interest in the study of microbial interrelationships.     

Phenol degradation by a stable aerobic consortium and its bacterial isolates

Summary An aerobic mixed culture removing phenol was developed and maintained with a biomass of 2900mg/l–3400 mg/l in a fed-batch reactor by feeding phenol 500 mg/l/day. The mixed culture (AS) consisted of two non-phenol bacteria and eight phenol degraders, of which 4 gram-negative rods and 4 gram-positive rods, that could remove about 96% of the fed phenol in 12 hours at 30°C±3°C. This paper also reports the stability of the consortium with respect to its constitution and phenol degradation.     

Influence of chemical surfactants on the biodegradation of crude oil by a mixed bacterial culture.

The effects of surfactant physicochemical properties, such as the hydrophile-lipophile balance (HLB) and molecular structure, on the biodegradation of 2% w/v Bow River crude oil by a mixed-bacterial culture were examined. Viable counts increased 4.6-fold and total petroleum hydrocarbon (TPH) biodegradation increased 57% in the presence of Igepal CO-630, a nonylphenol ethoxylate (HLB 13, 0.625 g/L). Only the nonylphenol ethoxylate with an HLB value of 13 substantially enhanced biodegradation. The surfactants from other chemical classes with HLB values of 13 (0.625 g/L) had no effect or were inhibitory. TPH biodegradation enhancement by Igepal CO-630 occurred at concentrations above the critical micelle concentration. When the effect of surfactant on individual oil fractions was examined, the biodegradation enhancement for the saturate and aromatic fractions was the same. In all cases, biodegradation resulted in increased resin and asphaltene concentrations. Optimal surfactant concentrations for TPH biodegradation reduced resin and asphaltene formation. Chemical surfactants have the potential to improve crude oil biodegradation in complex microbial systems, and surfactant selection should consider factors such as molecular structure, HLB, and surfactant concentration.     

Toxicity and degradation of metal-complexed cyanide by a bacterial consortium under sulfate-reducing conditions

Free cyanide at 1 mm decreased the initial sulfate reduction rate of a batch culture of granular sludge from 0.3 to 0.14 mmol d(-1) g(-1) SS (suspended solid), whereas 0.5 mm cyanide had a minimal effect (0.25 mmol d(-1) g(-1) SS). The order of toxicity of metal-complexed cyanides to the sludge was as follows: zinc-complexed cyanide (most toxic) > free cyanide = nickel-complexed cyanide > copper-complexed cyanide (least toxic), which also corresponds well with the order of the stability (dissociation) constants of the metal-cyanide complexes. A consortium degrading cyanide was enriched using nickel cyanide as the sole nitrogen source. This consortium completely removed 0.5 mm of nickel-complexed cyanide under sulfate-reducing conditions in 11 d. Analysis of clone library of 16S rRNA genes shows that the consortium was composed of three major phylotypes including Desulfovibrio.     

Biodegradation of crude oil across a wide range of salinities by an extremely halotolerant bacterial consortium MPD-M,immobilized onto polypropylene fibers

The bacterial consortium MPD-M, isolated from sediment associated with Colombian mangrove roots, was effective in the treatment of hydrocarbons in water with salinities varying from 0 to 180 g L(-1). Where the salinity of the culture medium surpassed 20 g L(-1), its effectiveness increased when the cells were immobilized on polypropylene fibers. Over the range of salinity evaluated, the immobilized cells significantly enhanced the biodegradation rate of crude oil compared with free-living cells, especially with increasing salinity in the culture medium. Contrary to that observed in free cell systems, the bacterial consortium MPD-M was highly stable in immobilized systems and it was not greatly affected by increments in salinity. Biodegradation was evident even at the highest salinity evaluated (180 g L(-1)), where biodegradation was between 4 and 7 times higher with immobilized cells compared to free cells. The biodegradation of pristane (PR) and phytane (PH) and of the aromatic fraction was also increased using cells immobilized on polypropylene fibers.     

Biodegradation of Tapis blended crude oil in marine sediment by a consortium of symbiotic bacteria

Biodegradation rate and the high molecular weight hydrocarbons are among the important concerns for bioremediation of crude oil. Inoculation of a non-oil-degrading bacterium as supplementary bacteria increased oil biodegradation from 57.1% to 63.0% after 10 days of incubation. Both the oil-degrading bacteria and the non-oil-degrading bacteria were isolated from Malaysian marine environment. Based on the 16S rDNA sequences, the oil-degrading bacteria was identified as Pseudomonas pseudoalcaligenes (99% similarity) while the non-oil-degrading bacterium was Erythrobacter citreus (99% similarity). E. citreus does not grow on crude oil enriched medium under present experimental condition but it withstands 5000 mg kg^?1 Tapis blended crude oil in sediment. Under optimal condition, the oil-degrading bacterium; P. pseudoalcaligenes, alone utilized 583.3 ± 3.8 mg kg^?1 (57.1%) at the rate of 3.97 × 10^?10 mg kg^?1 cell^?1 day^?1 Tapis blended crude oil from 1000 mg kg^?1 oil-contaminated sediment. Inoculation of E. citreus as the supplementary bacteria to P. pseudoalcaligenes enhanced biodegradation. The bacterial consortium degraded 675.8 ± 18.5 mg kg^?1 (63.0%) Tapis blended crude oil from the 1000 mg kg^?1 oil-contaminated sediment. Biodegradation rate of the bacterial consortium increased significantly to 4.59 × 10^?10 mg kg^?1 cell^?1 day^?1 (p = 0.02). Improvement of the oil degradation by the bacterial consortium was due to the synergetic reaction among the bacterial inoculants. There are two implications: (1) E. citreus may have a role in removing self-growth-inhibiting compounds of P. pseudoalcaligens. (2) P. pseudoalcaligenes degraded Tapis blended crude oil while E. citreus competes for the partially degraded hydrocarbons by P. pseudoalcaligenes. P. pseudoalcaligenes forced to breakdown more hydrocarbons to sustain its metabolic requirement. The bacterial consortium degraded 78.7% of (C₁₂–C₃₄) total aliphatic hydrocarbons (TAHs) and 74.1% of the 16 USEPA prioritized polycyclic aromatic hydrocarbons.     

Anaerobic degradation of phenol in batch and continuous cultures by a denitrifying bacterial consortium

Summary The anaerobic degradation of phenol under denitrifying conditions by a bacterial consortium was studied both in batch and continuous cultures. Anaerobic degradation was dependent on NO_f3 ^p– and concentrations up to 4 mm phenol were degraded within 2–5 days. During continuous growth in a fermenter, steady states could be maintained at eight dilution rates (D) corresponding to residence times between 12.5 and 50 h. Culture wash-out occurred at D=0.084 h^–1. The kinetic parameters obtained for anaerobic degradation of phenol under denitrifying conditions by the consortium were: maximam specific growth rate = 0.091 h^–1; saturation constant = 4.91 mg phenol/l; true growth yield = 0.57 mg dry wt/mg phenol; maintenance coefficient = 0.013 mg phenol/mg dry wt per hour. The Haldane model inhibition constant was estimated from batch culture data giving a value of 101 mg/l. The requirement of CO₂ for the anaerobic degradation of phenol with NO_f3 ^p– indicates that phenol carboxylation to 4-hydroxybenzoate was the first step of phenol degradation by this culture. 4-Hydroxybenzoate, proposed as an intermediate of phenol carboxylation under these conditions, was detected only in continuous cultures at very low growth rates (D=0.02 h^–1), but was never detected as a free intermediary metabolite either in batch or in continuous cultures. Correspondence to: N. Khoury     

Anaerobic degradation of p-cresol in batch and continuous cultures by a denitrifying bacterial consortium

Summary The anaerobic degradation of p-cresol under denitrifying conditions by a bacterial consortium was studied in batch and continuous cultures. Concentrations up to 3 mm were degraded within 5–6 days with 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde and 4-hydroxybenzoate as intermediates. Steady states could be maintained at only one dilution rate, D=0.04 h^–1. A further increase in the dilution rate to 0.0 8 h^–1 resulted in culture wash-out. An estimation of the Saturation constant was made (<1 mg/l), taking the maximum specific growth rate as 0.045 h^–1, thus yielding a value of 0.125 mg p-cresol/l. Correspondence to: N. Khoury     

降解吡啶复合菌系MD1的筛选、降解特性及代谢途径

【目的】为筛选吡啶高效降解复合菌系,促进高浓度吡啶废水的降解。本研究围绕吡啶降解复合菌系的筛选、降解特性及代谢途径,旨在获得吡啶高效降解复合菌系,为高浓度吡啶废水微生物降解及完全矿化提供理论依据和技术支撑。【方法】以吡啶为唯一碳氮源从某农药废水处理系统好氧活性污泥中筛选得到一个吡啶高效降解复合菌系MD1。采用16S rRNA高通量测序技术探究了MD1的群落结构及多样性,通过单因素实验考察了MD1的降解特性,利用气相色谱-质谱联用仪对MD1降解吡啶的代谢产物进行了初步检测与鉴定,推测吡啶可能的降解途径。【结果】结果显示,在温度30 ℃、pH 8.0、NaCl浓度0.1%的最佳条件下培养72 h,MD1对初始浓度1 400 mg/L的吡啶降解率为98.44%±0.27%。在属水平上,MD1主要由副球菌属(Paracoccus sp.)、布鲁氏菌属(unclassified_Brucellaceae)、无色杆菌属(Achromobacter sp.)等组成。由代谢产物检测结果初步推测MD1对吡啶的代谢途径为吡啶→烟酸→6-羟基烟酸→2,5-二羟基吡啶→N-甲酰基马来酰胺酸→马来酰胺酸→马来酸→CO₂+H₂O。【结论】研究筛选得到一个可高效降解吡啶、降解性能稳定的复合菌系MD1。解析了MD1的微生物组成多样性和群落结构,推测了MD1可能的代谢途径,研究结果丰富了吡啶降解微生物资源。     

Changes in mutagenicity during crude oil degradation by fungi

Two fungal strains, Cunninghamella elegans and Penicillium zonatum, that grow with crude oil as a sole carbon source were exposed to three crude oils that exhibit a range of mutagenic activity. At regular time intervals following fungal incubation with the various crude oils, extracts were tested for the presence of mutagenic activity using the spiral Salmonella assay. When the most mutagenic of the oils, Pennsylvania crude oil, was degraded by C. elegans or by P. zonatum, its mutagenicity was significantly reduced; corresponding uninoculated (weathered) controls of Pennsylvania crude remained mutagenic. West Texas Sour crude oil, a moderately mutagenic oil, exhibited little change in mutagenicity when incubated with either C. elegans or P. zonatum. Swanson River Field crude oil from Cook Inlet, Alaska is a slightly mutagenic oil that became more mutagenic when incubated with C. elegans; weathered controls of this oil showed little change in mutagenicity. Mycelial mat weights measured during growth on crude oils increased corresponding to the biodegradation of about 25% of the crude oil.     

Anaerobic degradation of sulphite evaporator condensate in a fixed-bed loop reactor by a defined bacterial consortium

Summary After elucidating the composition of an anaerobic bacterial enrichment culture treating sulphite evaporator condensate (SEC), an effluent in the pulp and paper industry, we built up stepwise a defined mixed culture to convert the organic constituents of SEC (acetate, methanol, furfural) to methane and CO₂. In batch cultures Desulfovibrio furfuralis and Methanobacterium bryantii degraded furfural in the absence of sulphate via inter-species H₂ transfer yielding 0.42 mol methane and 1.87 mol acetate/mol furfural degraded. When Methanosarcina barkeri was added to this diculture, acetate was also transformed to methane yielding 0.93 mol methane/mol acetate converted. This consortium (D. furfuralis, Methanobacterium bryantii and Methanosarcina barkeri) degraded furfural in continuous culture (fixed-bed loop reactor) to 92%, but the conversion of acetate was only 67%. The conversion of acetate could be further improved to 86% by adding 10 mm sulphate to the medium. This resulted in a space time yield of 10.9 g chemical oxygen demand (COD)/1 per day for the overall conversion. With a consortium consisting of M. barkeri, Methanobrevibacter arboriphilus, Methanosaeta concilii and D. furfuralis, a synthetic SEC could be degraded at a space time yield of 13.35 g COD/1 per day. This defined culture degraded all the constituents of SEC at an efficiency of almost 90% compared to an enrichment culture under identical conditions.Offprint requests to: U. Ney     

Synergistic degradation of linuron by a bacterial consortium and isolation of a single linuron-degrading variovorax strain

The bacterial community composition of a linuron-degrading enrichment culture and the role of the individual strains in linuron degradation have been determined by a combination of methods, such as denaturing gradient gel electrophoresis of the total 16S rRNA gene pool, isolation and identification of strains, and biodegradation assays. Three strains, Variovorax sp. strain WDL1, Delftia acidovorans WDL34, and Pseudomonas sp. strain WDL5, were isolated directly from the linuron-degrading culture. In addition, subculture of this enrichment culture on potential intermediates in the degradation pathway of linuron (i.e., N,O-dimethylhydroxylamine and 3-chloroaniline) resulted in the isolation of, respectively, Hyphomicrobium sulfonivorans WDL6 and Comamonas testosteroni WDL7. Of these five strains, only Variovorax sp. strain WDL1 was able to use linuron as the sole source of C, N, and energy. WDL1 first converted linuron to 3,4-dichloroaniline (3,4-DCA), which transiently accumulated in the medium but was subsequently degraded. To the best of our knowledge, this is the first report of a strain that degrades linuron further than the aromatic intermediates. Interestingly, the rate of linuron degradation by strain WDL1 was lower than that for the consortium, but was clearly increased when WDL1 was coinoculated with each of the other four strains. D. acidovorans WDL34 and C. testosteroni WDL7 were found to be responsible for degradation of the intermediate 3,4-DCA, and H. sulfonivorans WDL6 was the only strain able to degrade N,O-dimethylhydroxylamine. The role of Pseudomonas sp. strain WDL5 needs to be further elucidated. The degradation of linuron can thus be performed by a single isolate, Variovorax sp. strain WDL1, but is stimulated by a synergistic interaction with the other strains isolated from the same linuron-degrading culture.     

4-Chlorophenol degradation by a bacterial consortium: development of a granular activated carbon biofilm reactor

A bacterial consortium that can degrade chloro- and nitrophenols has been isolated from the rhizosphere of Phragmitis communis. Degradation of 4-chlorophenol (4-CP) by a consortium attached to granular activated carbon (GAC) in a biofilm reactor was evaluated during both open and closed modes of operation. During the operation of the biofilm reactor, 4-CP was not detected in the column effluent, being either adsorbed to the GAC or biodegraded by the consortium. When 4-CP at 100 mg l⁻¹ was fed to the column in open mode operation (20 mg g⁻¹ GAC total supply), up to 27% was immediately available for biodegradation, the rest being adsorbed to the GAC. Biodegradation continued after the system was returned to closed mode operation, indicating that GAC bound 4-CP became available to the consortium. Biofilm batch cultures supplied with 10–216 mg 4-CP g⁻¹ GAC suggested that a residual fraction of GAC-bound 4-CP was biologically unavailable. The consortium was able to metabolise 4-CP after perturbations by the addition of chromium (Cr VI) at 1–5 mg l⁻¹ and nitrate at concentrations up to 400 mg l⁻¹. The development of the biofilm structure was analysed by scanning electron microscopy and confocal laser scanning microscopy (CLSM) techniques. CLSM revealed a heterogeneous structure with a network of channels throughout the biofilm, partially occupied by microbial exopolymer structures. Received: 17 March 1999 / Received revision: 27 May 1999 / Accepted: 28 May 1999     

复合菌系降解纤维素过程中微生物群落结构的变化

为明确高效纤维素降解复合菌系降解过程中微生物群落结构的变化规律及关键的降解功能菌,利用该复合菌系对滤纸和稻秆进行生物处理,通过底物降解、微生物生长量、发酵液pH的变化情况,选择不同降解时期复合菌系提取的总DNA进行细菌16S rRNA基因扩增子高通量测序。通过分解特性试验确定在接种后培养第12、72、168 h分别作为降解初期、高峰期、末期。该复合菌系分别主要由1个门、2个纲、2个目、7个科、11个属组成。随着降解的进行,短芽胞杆菌属Brevibacillus、喜热菌属Caloramator的相对丰度逐渐降低;梭菌属Clostridium、芽胞杆菌属Bacillus、地芽胞杆菌属Geobacillus、柯恩氏菌属Cohnella的相对丰度逐渐升高;解脲芽胞杆菌属Ureibacillus、泰氏菌属Tissierella、刺尾鱼菌属Epulopiscium在降解高峰期时相对丰度最高;各时期类芽胞杆菌属Paenibacillus、瘤胃球菌属Ruminococcus的相对丰度无明显变化。上述11个主要菌属均属于厚壁菌门,具有嗜热、耐热、适应广泛pH、降解纤维素或半纤维素的特性。好氧型细菌是降解初期的主要优势功能菌,到中后期厌氧型细菌逐渐增多,并逐步取代好氧型细菌成为降解纤维素的主要细菌。     

Use of an internal standard in monitoring the bacterial degradation of crude oil.

Hexachloroethane is nonvolatile, insoluble in water, and apparently not toxic to or metabolized by bacteria. Its addition to cultures growing at the expense of crude oil thus provides an internal standard against which the rate of degradation of individual crude oil components can be conveniently and reproducibly measured.