Isolation and characterization of mesotrione-degrading Bacillus sp. from soil

Dissipation kinetics of mesotrione, a new triketone herbicide, sprayed on soil from Limagne (Puy-de-Dôme, France) showed that the soil microflora were able to biotransform it.Bacteria from this soil were cultured in mineral salt solution supplemented with mesotrione as sole source of carbon for the isolation of mesotrione-degrading bacteria. The bacterial community structure of the enrichment cultures was analyzed by temporal temperature gradient gel electrophoresis (TTGE). The TTGE fingerprints revealed that mesotrione had an impact on bacterial community structure only at its highest concentrations and showed mesotrione-sensitive and mesotrione-adapted strains. Two adapted strains, identified as Bacillus sp. and Arthrobacter sp., were isolated by colony hybridization methods.Biodegradation assays showed that only the Bacillus sp. strain was able to completely and rapidly biotransform mesotrione. Among several metabolites formed, 2-amino-4-methylsulfonylbenzoic acid (AMBA) accumulated in the medium. Although sulcotrione has a chemical structure closely resembling that of mesotrione, the isolates were unable to degrade it.     

Isolation and identification of beta-nitrostyrene from smoked chicken

Extraction and identification of beta-nitrostyrene from smoked chicken is described. Commercially obtained smoked chicken was homogenised and extracted with chloroform-methanol (2:1). The extracts were fractioned by silicic acid column chromatography. The purified extract using thin-layer chromatography was identified by a colour reaction (this qualitative method for some nitrated compounds is developed this time), high pressure liquid chromatography and gas chromatography/mass spectrometry. The concentration of beta-nitrostyrene in the smoked chicken was 94 ppb. The average recovery of added beta-nitrostyrene was 107%. However, the toxicity of this material was not tested in the present paper.     

膜生物反应器好氧反硝化菌的筛选及鉴定

从某膜生物反应器(MBR)污泥中筛选出反硝化活性较高的好氧反硝化细菌,进行了反硝化活性检测、菌种鉴定和作用机制探索。结果表明,共分离出6株好氧反硝化细菌,在较低的菌浓度(1×105个/mL)、DO为4.2~5.5mg/L的条件下启动脱氮反应,菌株F2、F4、F5在24、48h的总氮去除率分别超过40.29%、67.19%,硝酸盐氮去除率分别在64.21%、83.31%以上;经16SrDNA序列测序和比对,菌株F2、F4、F5分别与苍白杆菌属(Ochrobactrumsp.)、蜡状芽孢杆菌(Bacillus cereus)、布鲁菌属(Brucellasp.)的同源性最高;PCR扩增结果表明,菌株F2、F4、F5中均具有周质硝酸盐还原酶,这几种细菌很可能通过这种酶来实现好氧反硝化。     

Isolation and characterization of lead-tolerant Ochrobactrum intermedium and its role in enhancing lead accumulation by Eucalyptus camaldulensis

In this study, the potential of rhizospheric bacteria in promoting the growth and Pb accumulation by the woody plant Eucalyptus camaldulensis under hydroponic conditions was investigated for the first time. Three Pb-tolerant bacteria were isolated from the rhizosphere of E. camaldulensis grown in Pb-contaminated soils in the Bo Ngam Pb mine, Thailand. Based on analysis of partial 16S rRNA gene sequence, the three isolates were identified as Microbacterium paraoxydans BN-2, Ochrobactrum intermedium BN-3, and Bacillus fusiformis BN-4. Among these strains, O. intermedium BN-3 showed the highest tolerance to not only Pb but also Cd and Zn. After growth in the presence of Pb, the membranes of O. intermedium BN-3 cells exhibited an increase in unsaturated fatty acid levels but a decrease in fluidity. In hydroponic studies, inoculation of O. intermedium BN-3 significantly increased the biomass and Pb accumulation by E. camaldulensis compared to the uninoculated control. The results suggested the role of the natural rhizospheric bacteria localized to the root surface of E. camaldulensis in promoting Pb accumulation and plant growth. Our results indicate that O. intermedium BN-3 and other indigenous rhizospheric bacteria have the potential to improve the efficiency of phytoremediation of Pb-contaminated sites.     

Food-chain transfer of zinc from contaminated Urtica dioica and Acer pseudoplatanus L. to the aphids Microlophium carnosum and Drepanosiphum platanoidis Schrank

This study examines the food-chain transfer of Zn from two plant species, Urtica dioica (stinging nettle) and Acer pseudoplatanus (sycamore maple), into their corresponding aphid species, Microlophium carnosum and Drepanosiphum platanoidis. The plants were grown in a hydroponic system using solutions with increasing concentrations of Zn from 0.02 to 41.9 mg Zn/l. Above-ground tissue concentrations in U. dioica and M. carnosum increased with increasing Zn exposure (p < 0.001). Zn concentrations in A. pseudoplatanus also increased with solution concentration from the control to the 9.8 mg Zn/l solution, above which concentrations remained constant. Zn concentrations in both D. platanoidis and the phloem tissue of A. pseudoplatanus were not affected by the Zn concentration in the watering solution. It appears that A. pseudoplatanus was able to limit Zn transport in the phloem, resulting in constant Zn exposure to the aphids. Zn concentrations in D. platanoidis were around three times those in M. carnosum.     

The formation of trimethylarsine oxide from arsenobetaine by biodegradation with marine microorganisms

Bottom sediments collected from coastal waters were aerobically incubated in inorganic medium containing arsenobetaine as a carbon source. An arsenic compound converted from arsenobetaine during incubation was purified from the medium by ion exchange chromatography. The purified compound was identified to be trimethylarsine oxide by TLC, HPLC, FAB-MS and NMR.     

Evidence for the presence of arsenobetaine and another organoarsenical in shrimps

Hot base digestion of fish meal and shrimps in combination with sodium borohydride reduction produces only trimethylarsine (TMA). This treatment produces TMA in different amounts relative to the total arsenic content which is mainly of organic structure. These materials of marine origin yielded 94% and 69% resp. of the calculated amount of TMA. Arsenobetaine and arsenocholine were degraded under identical conditions and were found to produce only TMA (95% and 8% resp.). Furthermore, studies using thin-layer chromatography, electrophoresis and ion-exchange chromatography have confirmed the existence of one organoarsenic compound in fish meal and two organoarsenicals in shrimps. The major organoarsenic compound in fish meal was identified as arsenobetaine by masspectroscopy. In shrimps about $\text{2}\text{3}$ of the organoarsenic compounds was shown to be arsenobetaine and there is a strong indication for the presence of arsenocholine.     

Isolation and identification of 3-bromocarbazole-degrading bacteria

In this study, a bacterial strain, CH-1, capable of degrading 3-bromocarbazole (3-BCZ) was isolated from a polluted soil. Based on its physio-biochemical characteristics and 16S rRNA genes, strain CH-1 was identified as a Stenotrophomonas sp. Strain CH-1 was able to degrade 70% of 50 mg/L 3-BCZ within 8 d at pH 7.0 and 30°C in mineral salt medium (MSM). During the process, the main intermediate metabolite was identified as (2E, 4Z)-6-(2-amino-5-bromophenyl)-2-hydroxy-6-oxhexa-2, 4-dienoic by gas (2E, 4Z)-6-(2-amino-5-bromophenyl)-2-hydroxy-6-oxhexa-2,4-dienoic via gas chromatograph-mass spectrometry (GC-MS) analysis. The metabolite disappeared after 14 d, suggesting that the metabolite can also be degraded by strain CH-1. 3-BCZ is a new persistent organic pollutant. This is the first report of the biodegradation of 3-BCZ. The results indicated that strain CH-1 may be a promising bacterial candidate for the bioremediation of environments polluted with polyhalogenated carbazoles (PHCs).     

3株反硝化聚磷菌的分离与鉴定

通过烛缸法培养富集、分离,结合除磷试验、硝酸盐还原产气试验及异染颗粒和PHB颗粒染色辅助检验相结合的方法筛选,得到3株具有较高脱氮除磷效率的反硝化聚磷菌DNPA8, DNPA9和DNPA10。在富氮富磷培养基中培养48 h,各菌株的脱氮率均大于75%,除磷率均大于78%。采用多相分类的方法确定了3株反硝化聚磷菌的分类地位,DNPA8为嗜麦芽寡养单胞菌,DNPA9为水生丛毛单胞菌属首次发现的反硝化聚磷菌;DNPA10为约翰逊氏不动杆菌。该研究结果为富营养化水体的治理提供了有效的菌种资源。     

Measurement of arsenic compounds in littoral zone algae from the Western Mediterranean Sea. Occurrence of arsenobetaine

The determination of arsenic compounds in algae collected on the Catalan coast (Western Mediterranean) is reported. Ten algae species and the seagrass Posidonia oceanica were analyzed. Total arsenic in the samples was determined by microwave digestion and inductively coupled plasma mass spectrometry (ICPMS). Arsenic speciation in water extracts of samples was analyzed by liquid chromatography with both anionic and cationic exchange with ICPMS detection (LC-ICPMS). The total arsenic content of the algae samples ranged from 2.96 to 39.0mg As kg(-1). The following compounds were detected: arsenite (As(III)), arsenate (As(V)), methylarsonate (MA), dimethylarsinate (DMA), sulfonate sugar (SO3-sug), sulfate sugar (SO4-sug), phosphate sugar (PO4-sug), arsenobetaine (AB), arsenocholine (AC), trimethylarsine oxide (TMAO) and glycerol sugar (Gly-sug). The main arsenic species found were arsenosugars. Significant percentages of arsenobetaine (0.54 mg As kg(-1), 28% of the extractable arsenic and 0.39 mg As kg(-1), 18% of the extractable arsenic) were found in Ulva rigida and Enteromorpha compressa. These results are discussed in relation to the presence of epiphytes.     

Isolation and identification of intermediates from biodegradation of low chlorinated biphenyls (Delor-103)

Microorganism Pseudomonas species P2 metabolizes polychlorinated biphenyls (PCBs) and biphenyl, producing the whole spectrum of intermediates, among them coloured intermediates, which are suitable for the monitoring of PCBs degradation by optical sensors. Knowledge of chemical structures and conditions of development of colour metabolites is necessary for application of optical analytical methods. The main goal of this work was the isolation and identification of intermediates from the biodegradation of the mixture of low chlorinated biphenyls (Delor-103), which is based on the solid phase extraction (SPE) of the whole mixture using LiChrolut EN cartridges, then silylation of the extract as one way to the identification of one part of intermediates by GC-MS, and acetylation of the extract as a way for the further concentration and analysis of more polar chlorobiphenylols and chlorobiphenyldiols. The combination of SPE and following acetylation allows to obtain chlorobiphenylols and chlorobiphenyldiols as an almost pure fraction. The acetylation method could be also used instead SPE procedure with the same final concentration effect. Using the simulation mass spectrometry program, two new compounds, dihydrodihydroxytrichloro- and tetrahydrodihydroxytrichlorobiphenyl, as silylderivatives, were identified.     

一株鱼粉加工硫化氢恶臭气体脱除菌株的分离与鉴定

鱼粉是水产动物、及畜禽动物养殖饲料中无法替代的优质动物蛋白源。然而因加工原料新鲜度不断下降、腐败等原因,鱼粉加工过程中会释放出大量的硫化氢恶臭气体,导致周边环境的严重污染。取鱼粉厂污水沉淀池中的污泥,对脱硫微生物进行富集、分离、纯化、检测脱硫效率,得到一株高效脱硫菌株XJ-2,其脱硫效率达87.1%。通过菌株形态观察、生理生化实验、API 50CHB试剂条检测及16S rDNA测序鉴定,结果确定该菌为蜡状芽孢杆菌(Bacillus cereus)。研究菌株XJ-2生长特性,发现菌株对环境中的pH值耐受力较强,不耐高温,正好避开鱼粉厂休业期,且经生长曲线研究发现该菌的稳定期较长,具有良好的应用前景。此研究将为生物法脱除鱼粉加工恶臭气体的应用提供一定的理论基础。     

固废拆解场污染土壤中多环芳烃降解菌的筛选鉴定与降解特性

以腐植酸(HA) 溶液为吸附剂、从受多环芳烃污染的土壤中分离出来的降解菌制成为生物修复剂,以多环芳烃(PAHs)萘、菲、芘、荧蒽、苯并蒽、苯并芘为土壤污染物,对PAHs污染土壤进行修复实验。目的是筛选与分离吸附于HA的PAHs降解菌,研究HA与降解菌的协同效应对PAHs的降解效率的影响。用经过HA吸附的PAHs富集分离培养出1株高效降解菌株, 命名为Tzyx3,鉴定其为解脂耶氏酵母菌(Yarrowia lipolytica)。15 d后,土壤中萘、菲、芘、荧蒽、苯并蒽、苯并芘的降解率分别为90.7%、91.0%、74.7%、86.9%、84.7%和74.7%,表明Tzyx3和HA在PAHs污染土壤中存在协作关系,Tzyx3能够直接利用HA对土壤中的多环芳烃进行降解。     

一株微囊藻毒素降解菌的分离与鉴定

以水华蓝藻细胞中提取的微囊藻毒素为筛选物质,从太湖水华腐烂蓝藻中富集筛选出1株微囊藻毒素降解菌。该菌株革兰氏染色呈阴性,细胞细长杆状,菌落黄色,圆形,不透明,接触酶、氧化酶实验均呈阳性。16S rRNA基因序列的长度为1 416 bp(GenBank登录号为FJ976656)。系统发育树显示,该菌株与微嗜酸寡养单胞菌(Stenotrophomonas acidam-iniphila)的亲缘关系最近。通过菌株形态特征、生理生化特征和16S rRNA基因序列分析,将此菌株鉴定为微嗜酸寡养单胞菌,不同于已报道的微囊藻毒素降解菌属种。微囊藻毒素降解实验表明,该菌株5 d内将15.4 mg/L的微囊藻毒素完全降解,降解能力高于假单胞菌。     

AFB的活性污泥中发酵细菌的分离及其初步鉴定

从处理啤酒废水的厌氧流化床的活性污泥中分离纯化到了发酵细菌,并对其培养条件进行了研究,实验表明,该发酵细菌最佳培养温度为37～42℃,最佳培养时间为28 d,同时对其进行了生理生化和形态鉴定。将该发酵细菌反加到AFB反应器中,能有效降低反应器的启动时间,能提高反应器处理啤酒废水的处理效率约8%。     

Comparative embryotoxicity of a pentabrominated diphenyl ether mixture to common terns (Sterna hirundo) and American kestrels (Falco sparverius)

Concentrations of polybrominated diphenyl ethers (PBDEs) in Forster’s tern (Sterna forsteri) eggs from San Francisco Bay have been reported to range up to 63 μg g⁻¹ lipid weight. This value exceeds the lowest-observed-adverse-effect level (1.8 μg g⁻¹ egg wet weight; ∼32 μg⁻¹ lipid weight) reported in an embryotoxicity study with American kestrels (Falco sparverius). As a surrogate for Forster’s terns, common tern (Sterna hirundo) eggs were treated by air cell injection with corn oil vehicle (control) or a commercial penta-BDE formulation (DE-71) at nominal concentrations of 0.2, 2, and 20 μg g⁻¹ egg. As a positive control, kestrel eggs received vehicle or 20 μg DE-71 g⁻¹ egg. In terns, there were no effects of DE-71 on embryonic survival, and pipping or hatching success; however, treated eggs hatched later (0.44 d) than controls. Organ weights, organ-to-body weight ratios, and bone lengths did not differ, and histopathological observations were unremarkable. Several measures of hepatic oxidative stress in hatchling terns were not affected by DE-71, although there was some evidence of oxidative DNA damage (8-hydroxy-deoxyguanosine; 8-OH-dG). Although DE-71 did not impair pipping and hatching of kestrels, it did result in a delay in hatch, shorter humerus length, and reduced total thyroid weight. Concentrations of oxidized glutathione, reduced glutathione, thiobarbituric acid reactive substances, and 8-OH-dG in liver were greater in DE-71-treated kestrels compared to controls. Our findings suggest common tern embryos, and perhaps other tern species, are less sensitive to PBDEs than kestrel embryos.     

Isolation and identification of an anti-algal compound from Artemisia annua and mechanisms of inhibitory effect on algae

The goals of this work were to isolate and identify an anti-algal compound from extracts of Artemisia annua and study its mode of action on Microcystis aeruginosa. The anti-algal compound was isolated from the extracts using column chromatography and activity-guided fractionation methods. Artemisinin with strong anti-algal activity was identified by gas chromatography-mass spectrometry and ¹H Nuclear Magnetic Resonance. The EC₅₀ of artemisinin on M. aeruginosa was 3.2 mg L⁻¹. Artemisinin decreased the soluble protein content and increased the superoxide dismutase activity and ascorbic acid content of M. aeruginosa, but exerted no effect on soluble sugar content. The results suggested the mode of action of artemisinin on algae may primarily be the increasing level of reactive oxygen species in algae cells. The results of our research could aid in the development of new anti-algal substances and lead to further study of mechanisms of inhibitory effect on algae.     

2株乙酰甲胺磷降解菌的分离鉴定及降解特性研究

利用富集及驯化培养方法,从长期生产农药企业的废水处理系统、厂房周边污染的土壤和池水中,分离筛选出2株能够高效降解乙酰甲胺磷的菌株Y3、Y6。在形态特征和生理生化鉴定的基础上,对其16S rDNA序列进行了分析,并重点研究了它们对乙酰甲胺磷及其他农药的降解特性和抗性。结果表明,Y3、Y6分别为寡养单胞菌(Stenotrophomonas)和假单胞菌(Pseudomonas)。Y3、Y6在乙酰甲胺磷浓度分别为500 mg/L和1 000 mg/L,培养温度30℃,初始pH8,接种量为2.5%条件下,一周内可以将80%左右的乙酰甲胺磷矿化为磷酸根。外加葡萄糖及酵母膏对降解效率的研究表明,当酵母膏含量为1 g/L时,降解效果最理想;而外加葡萄糖的量,能相对抑制其对农药的降解。抗性实验显示,Y3、Y6均可在较高浓度的其他有机磷类及氨基甲酸酯类农药的普通培养基中生长,对其他农药的抗性也比较广泛。植物侵染实验显示,Y3、Y6对实验中的豆科、禾本科、十字花科及葫芦科植物不具备致病性,说明Y3、Y6环境安全性较强。     

甲醛降解细菌的分离鉴定和降解效果测定

从某化工厂污水处理系统的活性污泥中分离纯化得到一株能降解甲醛的革兰氏阳性菌,对其进行形态及生理生化特性测定,提取并纯化其基因组DNA,通过PCR扩增菌株的16S rDNA,测序结果表明,该菌株和苏云金芽孢杆菌的同源性为100%。在含1.2 μL/mL的甲醛的培养基上培养该菌36 h后,甲醛降解去除率达到72％以上。     

一株具有藻毒素降解和溶藻功能细菌的分离鉴定

从太湖水华腐烂蓝藻中富集筛选出一株编号A-8的细菌。该菌株菌落圆形、灰白色,革兰氏染色阴性,氧化酶阴性,接触酶阳性;分子系统进化树显示为溶血不动杆菌。根据以上结果,将此菌株鉴定为溶血不动杆菌。此外,对这株菌降解微囊藻毒素、不同生长时期的细菌的溶藻效果及溶藻专一性进行了研究。结果显示:该菌在3 d时间里将藻毒素降解了30.4%;细菌通过分泌胞外物质溶藻,细菌在衰退期时胞外分泌物的溶藻效果最好;该菌还对瘤状念珠藻、颤藻等9种蓝藻有溶藻作用。