Isolation, Characterization and Phylogenetic Analysis of an Aerobic Bacterium Capable of Degrading Bensulfuronmethyl

Summary An aerobic bacterium named strain BH was isolated from soil samples based on its bensulfuronmethyl-degrading characteristics using continuous enrichment cultures. The cells of the strain were non-motile, gram-positive short rods. Colonies formed on agar medium were round, smooth, sticky, white-yellow in colour and of butyrous consistency. Analyses of nutritional utilization in Biolog microplates, conventional phenotypic characteristics and 16S rRNA gene sequencing were consistent with assigning strain BH to the genus Brevibacterium. Growth of the cells and their ability to degrade bensulfuronmethyl were simultaneously monitored under different liquid medium conditions during 7 days of incubation. They degraded bensulfuronmethyl from 100 to 70.6 mg l⁻¹ in mineral M9 medium and exhibited more effective degradation in the presence of yeast extract, completely removing an initial concentration of 100 mg l⁻¹ and at best 80% of an initial concentration of 200 mg l⁻¹. Further studies are required to determine the potential use of the isolate in the disposal of bensulfuronmethyl residues in agriculture and industry.     

Isolation of a Bacterium Capable of Degrading Peanut Hull Lignin

Thirty-seven bacterial strains capable of degrading peanut hull lignin were isolated by using four types of lignin preparations and hot-water-extracted peanut hulls. One of the isolates, tentatively identified as Arthrobacter sp., was capable of utilizing all four lignin preparations as well as extracted peanut hulls as a sole source of carbon. The bacterium was also capable of degrading specifically labeled [¹⁴C]lignin-labeled lignocellulose and [¹⁴C]cellulose-labeled lignocellulose from the cordgrass Spartina alterniflora and could also degrade [¹⁴C]Kraft lignin from slash pine. After 10 days of incubation with [¹⁴C]cellulose-labeled lignocellulose or [¹⁴C]lignin-labeled lignocellulose from S. alterniflora, the bacterium mineralized 6.5% of the polysaccharide component and 2.9% of the lignin component.     

Isolation and Characterization of Bacteria Capable of Degrading Phenol and Reducing Nitrate Under Low-Oxygen Conditions

Nitrate-reducing bacteria capable of degrading phenol were isolated from natural and contaminated environments under low-oxygen conditions with nitrate-containing media, using phenol as a sole carbon and energy source. A total of 27 bacteria able to degrade phenol and reduce nitrate under low-oxygen conditions were isolated from all of the inoculum samples, regardless of previous phenol contamination. For all of these bacteria, oxygen was an essential requirement for phenol degradation. Nitrate reduction by 19 of the strains was insensitive to 10 mM sodium azide, and these strains were placed into the - and -subclasses of Proteobacteria and two were Gram-positive bacteria. To date, the order of Rhizobiales has hardly been reported to have an ability to degrade aromatic compounds. Interestingly, our study showed that all isolates that were placed into the -subclass of Proteobacteria are in the order of Rhizobiales. Furthermore, the genus Agrobacterium was isolated from most inoculum samples and one genus of Gram-positive bacteria, Staphylococcus, was also isolated. In the case of the remaining eight strains, nitrate reduction was inhibited by 10 mM sodium azide. Of these strains, seven were placed into the -subclass of Proteobacteria.     

敌敌畏降解菌的分离鉴定及降解特性研究

目的:从受有机磷农药污染的土壤中分离能降解DDVP的菌株,对其进行鉴定和降解特性研究.方法:采用DDVP为惟一碳源和能源的无机盐培养基,通过富集培养、平板划线分离得到一株优势菌,编号为DDW-1,采用形态学、生理生化和16S-rDNA序列分析对其进行鉴定,采用气相色谱测定菌株DDW-1对DDVP的降解能力,并进行底物广谱性测试和降解酶定位实验.结果:该菌株鉴定为甲基杆菌属(Methylobacterium sp.).降解特性试验结果表明,其最佳生长条件为温度28℃,初始pH为7.0,在该条件下,500mg·L-> DDVP经过DDW-1菌株代谢3d后,降解率达63.7%.结论:菌株DDW-1能降解DDVP,该菌株产胞内酶.     

Isolation,Characterization of a Strain Capable of Degrading Imazethapyr and Its Use in Degradation of the Herbicide in Soil

One strain of bacterium IM-4, capable of degrading imazethapyr (IMZT), was isolated from the IMZT-contaminated soil. The isolate was identified as Pseudomonas sp. according to its physiological characteristics, biochemical tests, and 16S rRNA gene phylogenetic analysis. This strain could utilize IMZT as the sole carbon and energy source. About 73.4% of the 50 mg l⁻¹ initially added IMZT was degraded after 7 days of inoculation with strain IM-4. This strain also showed the capability to degrade other imidazolinone herbicides such as imazapyr, imazapic, and imazamox. The inoculation strain IM-4 to soil treated with IMZT resulted in a higher degradation rate than in noninoculated soil regardless if the soil was sterilized or nonsterilized. Inoculation of strain IM-4 could also mitigate the phytotoxic effects of IMZT on the growth of maize.     

Comparison of Photocatalytic Oxidation and Ozonation in Degrading of Polycyclic Aromatic Hydrocarbons

Polycyclic aromatic hydrocarbons (PAHs) are hazardous pollutants. Photocatalytic oxidation (PCO) was used to degrade individual PAHs dissolved in aqueous solution and results were compared to those used for ozonation treatment of PAHs in aqueous phase and adsorbed on soil reported previously. Both PCO and ozonation showed high effectiveness and efficiency in degrading PAHs within only minutes of time. Ozonation showed higher performance in the degradation of PAHs in aqueous solution than in soil, whereas the treatment efficiency of PCO was higher than that of ozonation in degrading PAHs in aqueous solution. Both methods have advantages over each other but they have their own limitation(s) in practical application.     

Biodegradation of Aliphatic vs. Aromatic Hydrocarbons in Fertilized Arctic Soils

The objectives of this study were to (1) test a simple bioremediation treatment strategy in the Arctic and (2) examine the effect of fertilization on the degradation of aliphatic and aromatic hydrocarbons. The site is a coarse sand pad that once supported fuel storage tanks. Concentrations of diesel-range organics at the beginning of the study (July 1996) ranged from 250 to 860 mg/kg soil. Replicate field plots treated with fertilizer yielded final concentrations of 0, 50, 100, or 200 mg N/kg soil. Soil samples were collected three times during the thaw season and analyzed for physical and chemical properties, microbial populations and activities, and concentrations of semivolatile hydrocarbons. Soil pH and soil-water potentials declined as a result of fertilizer application. Addition of fertilizer significantly increased soil respiration potentials, but not the populations of microorganisms measured. Fertilizer addition also resulted in ∼50% loss of measured aliphatic and aromatic hydrocarbons in surface and subsurface soils. For fertilized plots, hydrocarbon loss was not related to the amount of fertilizer added. Losses of aliphatic hydrocarbons were attributed to biotic processes, whereas losses of aromatic hydrocarbons likely were a result of both biotic and abiotic processes.     

Isolation and Characterization of a Subsurface Bacterium Capable of Growth on Toluene, Naphthalene, and Other Aromatic Compounds

A bacterium, designated F199, utilized toluene, naphthalene, dibenzothiophene, salicylate, benzoate, p-cresol, and all isomers of xylene as a sole carbon and energy source. This bacterium was isolated from Middendorf sediments, a Cretaceous age formation that underlies the Southeast Coastal Plain in South Carolina, at a depth of approximately 410 m. F199 is a gram-positive, irregular-shaped bacterium that has a varied cell morphology that is dependent on culture medium type and growth stage. F199 required microaerobic conditions (40 to 80 μM O₂) for growth on hydrocarbons, glucose, acetate, and lactate in mineral salts medium but not for growth on rich media. [¹⁴C]naphthalene mineralization by F199 was induced by either naphthalene or toulene; however, [¹⁴C]toluene mineralization by this strain was induced by toluene but not naphthalene. F199 was also found to harbor two plasmids larger than 100 kb. Restricted F199 plasmid and genomic DNA did not hybridize with toluene (pWW0) or naphthalene (NAH7) catabolic plasmid DNA probes. The presence in the Middendorf formation of bacteria with the capacity for degrading a variety of aromatic compounds suggests that indigenous microorganisms may have potential for in situ degradation of organic contaminants.     

Bacterial Diversity of a Consortium Degrading High-Molecular-Weight Polycyclic Aromatic Hydrocarbons in a Two-Liquid Phase Biosystem

High-molecular-weight (HMW) polycyclic aromatic hydrocarbons (PAHs) are pollutants that persist in the environment due to their low solubility in water and their sequestration by soil and sediments. Although several PAH-degrading bacterial species have been isolated, it is not expected that a single isolate would exhibit the ability to degrade completely all PAHs. A consortium composed of different microorganisms can better achieve this. Two-liquid phase (TLP) culture systems have been developed to increase the bioavailability of poorly soluble substrates for uptake and biodegradation by microorganisms. By combining a silicone oil–water TLP system with a microbial consortium capable of degrading HMW PAHs, we previously developed a highly efficient PAH-degrading system. In this report, we characterized the bacterial diversity of the consortium with a combination of culture-dependent and culture-independent methods. Polymerase chain reaction (PCR) of part of the 16S ribosomal RNA gene (rDNA) sequences combined with denaturing gradient gel electrophoresis was used to monitor the bacterial population changes during PAH degradation of the consortium when pyrene, chrysene, and benzo[a]pyrene were provided together or separately in the TLP cultures. No substantial changes in bacterial profiles occurred during biodegradation of pyrene and chrysene in these cultures. However, the addition of the low-molecular-weight PAHs phenanthrene or naphthalene in the system favored one bacterial species related to Sphingobium yanoikuyae. Eleven bacterial strains were isolated from the consortium but, interestingly, only one—IAFILS9 affiliated to Novosphingobium pentaromativorans—was capable of growing on pyrene and chrysene as sole source of carbon. A 16S rDNA library was derived from the consortium to identify noncultured bacteria. Among 86 clones screened, 20 were affiliated to different bacterial species–genera. Only three strains were represented in the screened clones. Eighty-five percent of clones and strains were affiliated to Alphaproteobacteria and Betaproteobacteria; among them, several were affiliated to bacterial species known for their PAH degradation activities such as those belonging to the Sphingomonadaceae. Finally, three genes involved in the degradation of aromatic molecules were detected in the consortium and two in IAFILS9. This study provides information on the bacterial composition of a HWM PAH-degrading consortium and its dynamics in a TLP biosystem during PAH degradation.     

氯苯降解菌的筛选鉴定及降解特性研究

本文采集化工厂排污口的污泥样品, 在含有氯苯为唯一碳源的基本培养基中, 先后分离筛选出7株能够降解氯苯的微生物菌株。通过对分离菌株的16S rRNA基因序列进行分析, 发现其中5株细菌分别属于放线菌目的考克氏菌属(KD139)、红球菌属(KD140和KD142)和节杆菌属(KD230和KD232), 1株细菌属于杆菌目的芽胞杆菌d属(KD178), 另外1株细菌属于黄色单孢菌目的寡食单胞菌属(KD237); 同时我们构建了系统进化树, 确定分离菌株的相对进化地位。本文还利用气相色谱方法, 对分离菌株降解氯苯的能力进行了初步分析, 其中寡食单胞菌KD237降解氯苯能力最高, 24 h内氯苯分解率达60.78%。     

氯嘧磺隆降解菌株LW-3的分离及生物学特性研究

从长期受氯嘧磺隆污染的土壤中分离到一株氯嘧磺隆高效降解菌株,命名为LW-3,菌株LW-3可以氯嘧磺隆为唯一氮源生长,接种量为2%时,50 mg/L氯嘧磺隆经过7 d,降解率达70%～80%.生理生化实验和16S rDNA序列同源性分析,将菌株LW-3归属于假单胞茵属(Pseudomonas sp.);菌株的适宜降解条件为;温度30℃～35℃,pH 6.5～7.2,pH对菌株LW-3降解氯嘧磺隆有明显地影响.     

Biodegradation of Aliphatic and Aromatic Hydrocarbons by Nocardia sp. H17-1

We investigated the biodegradation of hydrocarbon components by Nocardia sp. H17-1 and the catabolic genes involved in the degradation pathways of both aliphatic and aromatic hydrocarbons. After 6 days of incubation, the aliphatic and aromatic fractions separated from Arabian light oil were degraded 99.0 ± 0.1% and 23.8 ± 0.8%, respectively. Detection of the catabolic genes involved in the hydrocarbon degradation indicated that H17-1 possessed the alkB genes for n-alkane biodegradation and catA gene for catechol 1,2-dioxygenase. However, H17-1 had neither the C23O gene for the degradation of aromatic hydrocarbons nor the catechol 2,3-dioxygenase activity. The investigation of the genes involved in the biodegradation of hydrocarbons supported the low degradation activity of H17-1 on the aromatic fractions.     

一株高效纤维素降解菌株的分离鉴定及其酶学性质

从黄浦江淀山湖的水底沉积物中筛选分离得到一株产纤维素酶的菌株。经细菌形态观察,生理生化实验并结合16SrRNA序列分析,鉴定该菌为蜡状芽孢杆菌(Bacillus cereus),同时发现其在系统发育树中处于一个独立的分支,推测该菌为Bacillus属中一个新的亚种。对该菌株产酶及酶活特性进行了初步研究,发现纤维素酶的产生与细菌的生长密切相关。该菌株在37°C,pH7.0的条件下,发酵66h后纤维素酶活达到最高值4.58U/mL。     

Isolation and Characterization of a Pseudomonas Oleovorans Degrading the Chloroacetamide Herbicide Acetochlor

To date, no pure bacterial cultures that could degrade acetochlor have been described. In this study, one strain of microorganism capable of degrading acetochlor, designated as LCa2, was isolated from acetochlor-contaminated soil. The strain LCa2 is Pseudomonas oleovorans according to the criteria of Bergey’s manual of determinative bacteriology and sequence analysis of the partial 16S rRNA gene. Optimum growth temperature and pH were 35 °C and 8.0, respectively. The strain could degrade 98.03% of acetochlor treated at a concentration of 7.6 mg l⁻¹ after 7 days of incubation and could tolerate 200 mg l⁻¹ of acetochlor. When the acetochlor concentration became higher, the degradation cycle became longer. The acetochlor biodegradation products were identified by GC–MS based on mass spectral data and fragmentation patterns. The main plausible degradative pathways involved dechlorination, hydroxylation, N-dealkylation, C-dealkylation and dehydrogenation.     

A Comparison of Isolation Methods for Black Fungi Degrading Aromatic Toxins

Mycopathologia - The prevalence of black fungi in the order Chaetothyriales has often been underestimated due to the difficulty of their isolation. In this study, three methods which are often used...     

三唑磷降解菌株GS-1的分离鉴定及其降解特性的研究

从有机磷农药污水处理池污泥中分离到一株能高效降解三唑磷的菌株GS-1, 通过生理生化实验和16S rDNA序列同源性分析, 将该菌株鉴定为Diaphorobacter sp.。菌株GS-1能以三唑磷为唯一碳源生长, 能在12 h内降解100 mg/L的三唑磷至检测不出的水平。菌株GS-1降解三唑磷的过程中会产生中间代谢产物苯唑醇(1-苯基-3-羟基-1,2,4-三唑), 36 h后苯唑醇被完全转化。菌株GS-1降解三唑磷的最适pH值为8.0, 最适温度为30°C, 且对杀螟硫磷、辛硫磷、毒死蜱和甲基对硫磷     

Isolation and Screening of Black Fungi as Degraders of Volatile Aromatic Hydrocarbons

Black fungi reported as degraders of volatile aromatic compounds were isolated from hydrocarbon-polluted sites and indoor environments. Several of the species encountered are known opportunistic pathogens or are closely related to pathogenic species causing severe mycoses, among which are neurological infections in immunocompetent individuals. Given the scale of the problem of environmental pollution and the phylogenetic relation of aromate-degrading black fungi with pathogenic siblings, it is of great interest to select strains able to mineralize these substrates efficiently without any risk for public health. Fifty-six black strains were obtained from human-made environments rich in hydrocarbons (gasoline car tanks, washing machine soap dispensers) after enrichment with some phenolic intermediates of toluene and styrene fungal metabolism. Based on ITS sequencing identification, the majority of the obtained isolates were members of the genus Exophiala. Exophiala xenobiotica was found to be the dominant black yeast present in the car gasoline tanks. A higher biodiversity, with three Exophiala species, was found in soap dispensers of washing machines. Strains obtained were screened using a 2,6-dichlorophenol-indophenol (DCPIP) assay, optimized for black fungi, to assess their potential ability to degrade toluene. Seven out of twenty strains tested were able to use toluene as carbon source.     

一株高分子量多环芳烃降解菌的筛选、鉴定及降解特性研究

采用富集培养方法从多环芳烃污染土壤中筛选分离得到1株能以苯并[a]芘(B[a]P)为唯一碳源和能源生长的菌株.形态特征观察和16S rDNA序列分析结果表明,该菌株为副球菌属(Paracoccus sp.),编号为HPD-2.HPD-2在3.0 mg/L的B[a]P液体培养基中生长较慢,培养5 d后B[a]P的降解率为89.7%.同时,该菌株对四环的芘和荧蒽也具有较好的降解能力,培养7 d后芘和荧蒽的降解率分别达到47.2%和84.5%.可见,该菌株对高分子量PAHs具有很好的降解潜力.     

Isolation and Characterization of Antagonistic Bacillus Strains Capable to Degrade Ethylenethiourea

In this study, more than 150 bacteria showing antagonistic properties against bacterial and fungal pathogens of the tomato plant were isolated and characterized. The most efficient agents against these phytopathogenic microorganisms belong to the genus Bacillus: the best biocontrol isolates were representatives of Bacillus subtilis, B. mojavensis and B. amyloliquefaciens species. They intensively produced fengycin or/and surfactin depsipeptide antibiotics and also proved to be excellent protease secretors. It was proved, that the selected strains were able to use ethylenethiourea (ETU) as sole nitrogen source. These antagonistic and ETU-degrading Bacillus strains can be applied as biocontrol and also as bioremediation agents.