Characterization of a salt-tolerant bacterium Bacillus sp. from a membrane bioreactor for saline wastewater treatment

High salt concentrations can cause plasmolysis and loss of activity of cells, but the salt-torlerant bacterium can endure the high salt concentrations in wastewater. In this research 7 salt-torlerant bacteria, which could survive in dry powder products and could degrade organic contaminants in saline wastewater, were isolated from a membrane bioreactor. The strain NY6 which showed the fastest growth rate, best property for organic matter degradation and could survive in dry powder more than 3 months was selected and characterized. It was classified as Bacillus aerius based on the analysis of the morphological and physiological properties as well as the 16 S rRNA sequence and Neigh borjoining tree. The strain NY6 could survive in the salinity up to 6% and the optimal growth salinity is2%; it belongs to a slightly halophilic bacterium. The capability of its dry powder products for COD removal was 800 mg COD/(g·day) in synthesized saline wastewater with salinity of 2%. According to salt-tolerant mechanism research, when the salinity was below 2%, the stain NY6 absorbed K+and Na+to maintain osmotic equilibrium, and when the salinity was above 2%, the NY6 kept its life by producing a large amount of spores.     

异养硝化细菌Bacillus sp. LY脱氮性能研究

研究了异养硝化细菌Bacillus sp. LY的脱氮性能.结果表明,Bacillus sp. LY是1株具有脱氮能力的异养硝化细菌.在NH⁺₄-N浓度分别为40、80和120 mg/L 3种情况下,120 h反应后,氨氮的去除率分别是100%、85.7%、73.7%,总氮的去除率分别是76.6%、53.4%、64.8%,在菌液初始浓度相同的情况下,随着NH⁺₄-N浓度的增加,细菌的硝化速率以及脱氮速率呈现下降的趋势.有机物浓度是影响Bacillus sp. LY脱氮性能的重要因素,低的有机物浓度会阻碍细菌脱氮性能的发挥,中的有机物浓度会促进细菌脱氮性能的发挥,使体系的脱氮效果达到最佳,高的有机物浓度并不能再次提升细菌的脱氮性能.在Bacillus sp. LY作用下,有机氮经过氨化作用生成氨氮,通过2条可能的途径转化为氮气.1条途径是氨氮先硝化生成亚硝酸盐与硝酸盐,然后反硝化生成氮气.另1条途径是氨氮被氧化生成羟胺,然后脱氢生成氧化亚氮并进一步转化为氮气.这些研究可为开发新型高效生物脱氮工艺提供参考.     

耐砷芽孢杆菌对As3+的吸附性能与机制研究

为了给砷污染水体的微生物修复提供理想材料和必要的理论依据,从湖南某矿区筛选分离得到一株高耐砷菌株,利用16S rDNA基因测序分析对其进行鉴定.同时,采用单因素试验研究了耐性菌吸附As³⁺的影响因素及规律,通过研究等温吸附属性、吸附动力学和热力学属性,分析亚细胞赋存特性,并结合扫描电镜（SEM）、傅里叶红外光谱（FTIR）等技术手段,初步探讨了吸附发生的可能机理.结果表明,通过形态学、生理生化及分子鉴定,初步鉴定为芽孢杆菌属（Bacillus sp.）,命名为Bacillus sp. strain AsT4.该菌株固体平板培养对砷的耐受阈值为40 mmol·L^-1;菌株最适生长条件为温度35℃、pH=7.0、NaCl浓度5 g·L^-1、转速180 r·min^-1;芽孢杆菌湿菌体吸附As³⁺的优化条件为：温度35℃,溶液pH=6.0,培养时间36 h,As³⁺初始浓度10 mg·L^-1,菌量1.5 mL,此条件下的去除率为69.3%.干菌体吸附As³⁺的优化条件为：温度35℃,pH=7.0,吸附时间90 min,As³⁺初始浓度10 mg·L^-1,投加量0.5 g·L^-1,此条件下的吸附率为72.8%.无论是湿菌体还是干菌体,Langmuir等温吸附模型和拟二级动力学方程能更好地描述耐砷芽孢杆菌AsT4对As³⁺的吸附过程,并且是自发、墒增的吸热过程.耐性菌对As³⁺的生物吸附和累积以胞内累积和细胞壁富集为主,菌粉表面的羧基、羟基、胺基等活性基团可能在吸附过程中起主要作用.     

畜禽粪便源芽孢杆菌多样性及对铜耐受性研究

芽孢杆菌制剂广泛应用于畜禽养殖和粪污处理等领域,铜作为饲料添加剂用于畜禽养殖,导致畜禽粪便中大量残留,在此压力下芽孢杆菌等会产生铜耐受性,由于芽孢杆菌的抗逆性,这种耐受性可由可移动遗传元件介导在环境中传播.本研究采集了多个规模化养殖场的牛粪、鸡粪和猪粪,从粪便中分离芽孢杆菌,通过PCR技术、最小抑菌浓度测定和全基因组测序方法,了解畜禽粪便中芽孢杆菌对铜耐受性.研究发现：(1)分离出芽孢杆菌属23种芽孢杆菌共235株.分离数量前三分别是枯草芽孢杆菌(Bacillus subtilis)77株,占总数的32.77%;蜡样芽孢杆菌(B.cereus)31株,占比13.19%;贝莱斯芽孢杆菌(B.velezensis)28株,占比11.91%.(2)芽孢杆菌对铜耐受表型和基因型研究发现,铜对芽孢杆菌的MIC集中在200～400 mg·L^-1,占总数235株的77.87%(n=183),铜耐受能力较高的菌株能够达到500～600 mg·L^-1,占总数的16.17%(n=38),5株MIC达到600 mg·L^-1的菌株全部来自于猪粪...     

2株芽孢杆菌利用不同碳源生产絮凝剂的研究

探讨了在纯培养和混合培养的条件下,2株芽孢杆菌F2和F6生产高效絮凝剂过程中对碳源的利用情况.研究表明,多种分子量在200以下的有机物能够作为碳源用于高效絮凝剂的生产,包括:单糖、双糖、醇、有机酸和酯类等,具有一定的普适性.发现利用废糖蜜、生物制氢废液和纤维素糖化液等廉价的生物质废料能够生产出性能良好的生物絮凝剂.     

Methylopila sp.SD-1与大豆联合修复苯磺隆污染土壤

分离得到1株苯磺隆降解菌株SD-1,根据表型、生理生化特征及16S rRNA基因序列分析,将其鉴定为Methylopila sp.SD-1,为首次报道能够降解苯磺隆的Methylopila属菌株,其在4d内完全降解50mg/L苯磺隆,最适降解温度、pH值分别为30℃和7.0,降解中间产物对大豆的毒性显著降低.大豆根系分泌物能促进菌株SD-1的生长,培养5d,菌株SD-1的数量由1.0×10⁷CFU/mL增至6.7×10⁷CFU/mL.分泌物中含有16种氨基酸,菌株SD-1对其中的Asp、Glu和Phe表现出明显的趋化性.接种菌悬液至苯磺隆污染土壤（3mg/kg）并种植大豆幼苗,培养4d,菌株SD-1依赖趋化性向大豆根系运动并定殖,存活率提高,根际土壤中苯磺隆的降解率相较于未种植大豆的处理提高36.0%.     

Methylopila sp.SD-1与大豆联合修复苯磺隆污染土壤

分离得到1株苯磺隆降解菌株SD-1,根据表型、生理生化特征及16S rRNA基因序列分析,将其鉴定为Methylopila sp.SD-1,为首次报道能够降解苯磺隆的Methylopila属菌株,其在4d内完全降解50mg/L苯磺隆,最适降解温度、pH值分别为30℃和7.0,降解中间产物对大豆的毒性显著降低.大豆根系分泌物能促进菌株SD-1的生长,培养5d,菌株SD-1的数量由1.0×10⁷CFU/mL增至6.7×10⁷CFU/mL.分泌物中含有16种氨基酸,菌株SD-1对其中的Asp、Glu和Phe表现出明显的趋化性.接种菌悬液至苯磺隆污染土壤（3mg/kg）并种植大豆幼苗,培养4d,菌株SD-1依赖趋化性向大豆根系运动并定殖,存活率提高,根际土壤中苯磺隆的降解率相较于未种植大豆的处理提高36.0%.     

城市尾水氮代谢过程中芽孢杆菌对微藻作用机制

为了对城市尾水深度脱氮,提高地表水体质量,分别利用微藻（Z）、芽孢杆菌（Y）和微藻芽孢杆菌（ZY）对城市尾水进行脱氮处理与氮代谢菌群特征研究,结果表明,Z和ZY对城市尾水中氨氮的去除效果较好,2组降解率都达到了95%以上.Z对亚硝态氮去除效果最好,芽孢杆菌与微藻的共同作用在氮循环反应中亚硝态氮转化为硝态氮过程发挥出较为强大的稳定效果.芽孢杆菌可以有效去除硝态氮,并可以提高微藻对硝态氮的去除效率.ZY菌对城市尾水中硝态氮的去除效果最好,其降解率高达99%以上,几乎完全去除城市尾水中的硝态氮.在Z中样本数占比较高的菌群分别为Chroococcidiopsis_PCC_7203 （24.38%）、uncultured_bacterium-g_norank_f_A4b （23.65%）、Exiguobacteriu （7.09%）、Leptolyngbya_PCC-6306 （9.41%）和Bacillus （1.99%）.在ZY中样本数占比较高的菌群分别为Brevibacillus （22.94%）、Clostridium（8.78%）和Bacillus （4.88%）,Chroococcidiopsis_PCC_7203 样本数占比仅为7.84%,远远低于Z样本数所占比例.微藻可以很好去除系统中氨氮,芽孢杆菌与微藻联用具有较好的氨氮和硝态氮去除效果,芽孢杆菌具有抑制微藻过度增长作用,防止水体富营养化和黑臭发生.研究可为城市尾水深度处理,防止地表水体富营养化提供数据支持.     

一株芽孢杆菌在低氮源浓度培养基中的生长与氨氮去除特性

对菌株YB3进行了16S rRNA基因序列进化分析,并分别以NH_4Cl、NaNO_2、NaNO_3、尿素和蛋白胨为单一氮源,配制了5种低氮源浓度培养基,研究YB3在与养殖水体相近营养水平条件下的生长与氨氮去除特性.结果显示,菌株YB3属于蜡样芽孢杆菌(Bacullis cereus),在5种培养基中均能够生长,菌悬液(吸光度OD600为1.0)接种量为1.0%(v/v)时,OD600由0.010增长到0.100~0.117.在NH_4Cl培养基中,YB3的氨氮去除速率为1.23 mg·L~(-1)·d~(-1),去除率为93.5%.在尿素、蛋白胨等有机氮源培养基中,YB3将首先导致氨氮的积累,累积倍数分别为51.69和3.38,之后开始去除,去除速率为1.56和0.29 mg·L~(-1)·d~(-1),去除率为93.7%和26.8%.结果也表明,提高YB3接种量至8.0%(v/v),可以使蛋白胨培养基氨氮累积倍数下降至2.02,去除速率提高至1.07 mg·L~(-1)·d~(-1),去除率最终达到98.4%.NaNO_2和NaNO_3培养基中均未检测到氨氮,而NH_4Cl、尿素和蛋白胨培养基中也未检测到NO_2~--N和NO_3~--N,表明YB3的硝化、亚硝化和反硝化作用均不强烈,去除氨氮的同时将不会造成NO_2~--N和NO_3~--N等的大量积累.本文为菌株YB3在养殖水体调控与净化中的应用研究提供了实验基础和理论支持.     

Evaluation of Bacillus sp. MZS10 for decolorizing Azure B dye and its decolorization mechanism

To evaluate decolorization and detoxification of Azure B dye by a newly isolated Bacillus sp. MZS10 strain, the cultivation medium and decolorization mechanism of the isolate were investigated. The decolorization was discovered to be dependent on cell density of the isolate and reached 93.55%(0.04 g/L) after 14 hr of cultivation in a 5 L stirred-tank fermenter at 2.0 g/L yeast extract and 6.0 g/L soluble starch and a small amount of mineral salts. The decolorization metabolites were identified with ultra performance liquid chromatography-tandem mass spectroscopy(UPLC-MS). A mechanism for decolorization of Azure B was proposed as follows: the C=N in Azure B was initially reduced to –NH by nicotinamide adenine dinucleotide phosphate(NADPH)-dependent quinone dehydrogenase, and then the –NH further combined with –OH derived from glucose to form a stable and colorless compound through a dehydration reaction. The phytotoxicity was evaluated for both Azure B and its related derivatives produced by Bacillus sp. MZS10 decolorization, indicating that the decolorization metabolites were less toxic than original dye. The decolorization efficiency and mechanism shown by Bacillus sp. MZS10 provided insight on its potential application for the bioremediation of the dye Azure B.     

降解芘的分枝杆菌M11的分离鉴定和降解特性

从多环芳烃污染的土壤中分离到1株能高效降解四环芳烃芘的放线菌M11,经形态观察、生理生化和16S rDNA鉴定,属于分枝杆菌属(Mycobacterium sp.).菌株M11能以菲、蒽、荧蒽和芘为唯一碳源生长,在含芘50、100和200 mg/L的无机盐液体培养基中培养16 d降解率分别达到76.9%、91.8%和79.23%.菌株M11对芘的降解具有较广泛的pH范围,在芘浓度100 mg/L,pH为5～9的液体条件下,均可生长.根据已报道的芘降解菌的双加氧酶同源序列设计引物,PCR扩增出编码双加氧酶大亚基和小亚基的基因片段,序列分析表明与已知降解芘的分枝杆菌的双加氧酶基因具有高度同源性.     

低营养水体中芽孢杆菌降解有机氮的研究

芽孢杆菌具有降解有机氮的功能,但在养殖水体等低营养水体中,其降解效果可能受到影响.为研究低营养水体中有机氮的降解情况,通过模拟凡纳滨对虾中间培育过程配制低营养水体,分别接种芽孢杆菌NT9和YB3(NT9水体和YB3水体),然后研究水体中微生物的生长与有机氮的降解情况,并构建数学模型进行分析.结果显示,起始接种量为10×10~5 cfu·mL~(-1)时,NT9水体总菌量呈下降趋势,平均为(3.46×10~5±2.39×10~5) cfu·mL~(-1),YB3水体总菌量则上升到(25.43×10~5±8.84×10~5) cfu·mL~(-1),但均高于未接种的对照水体.NT9水体和YB3水体的有机氮降解率显著高于对照水体(p0.05),分别提高50.28%和119.41%,降解速率也分别提高65.22%和121.74%.对照水体、NT9水体和YB3水体单位菌量的有机氮降解效率分别为1.238、1.649和1.904 mg·L~(-1),降解模型分别为y=-6.40+1.39x_1+1.45x_2、y=2.11+8.21x_3-0.64x_4-1.26x_1x_3-0.32x_2x_4和y=1.73+6.11x_2(x_1、x_2、x_3、x_4分别表示总菌量、总菌增量、有机氮含量和时间).研究表明,在低营养水体中接种芽孢杆菌有利于有机氮的降解,但不同的菌株具有不同的降解模式,菌株YB3为能够适应低营养水平、增殖能力较强的菌株,可以更有效地促进有机氮的降解,提高降解效率.     

高量秸秆还田配施芽孢杆菌对沙化土壤细菌群落及肥力的影响

探讨高量秸秆还田模式协同配施芽孢杆菌等功能菌群对沙化土壤的培肥潜能,解析土壤碳氮磷组分和菌群功能活性的变化特征,可为高效提升沙化土壤肥力提供依据.采用随机区组试验,设置秸秆不还田（CK）、高量还田梯度分别为6.00 kg ·m^-2（ST1）、12.00 kg ·m^-2（ST2）、24.00 kg ·m^-2（ST3）、6.00 kg ·m^-2+芽孢杆菌（SM1）、12.00 kg ·m^-2+芽孢杆菌（SM2）和24.00 kg ·m^-2+芽孢杆菌（SM3）这7个处理组.利用16S rRNA高通量测序技术分析细菌多样性和群落结构,土壤农化分析方法测定土壤化学性质.结果发现：①高量秸秆还田协同配施芽孢杆菌显著降低土壤细菌群落α多样性;②高量秸秆还田单一模式显著富集变形菌门,降低放线菌门的相对丰度,配施芽孢杆菌对细菌群落结构变异性的影响更显著,在属水平上表现为假单胞菌、罗河杆菌和芽孢杆菌等有益菌属的相对丰度显著增加;③基于FAPROTAX的功能预测发现：高量秸秆还田配施芽孢杆菌显著提高了土壤菌群对有机物质的分解潜力和氮组分转化潜力;④与对照相比较,高量秸秆还田配施芽孢杆菌显著提升了ω（SOM）、ω（TP）和ω（AP）,分别提高了31.20~32.75 g ·kg^-1、0.11~0.18 g ·kg^-1和29.69~35.09 mg ·kg^-1.因此,高量秸秆还田配施芽孢杆菌可显著提升沙化土壤有机质与磷组分含量、细菌功能活性与有益菌属丰度,对旱区沙化中低产田地力快速提升具有重要意义.     

嗜盐高效降酚菌株Halomonas sp.H17的筛选及降解苯酚特性

从巴丹吉林沙漠盐湖沉积物中分离获得1株在高盐环境下高效降解苯酚菌H17.分析了H17生理生化特性、16S rDNA基因序列、苯酚降解特性及动力学,结果表明,H17属于盐单胞菌属(Halomonas sp.),能在0～20%的盐度下有效降解苯酚,每升外加适量的碳源(葡萄糖浓度0.8 g)和复合氮源(KNO_3 1 g、NH_4Cl 5 g、酵母提取物0.2 g和胰蛋白胨0.2 g)能够促进H17的生长及降解苯酚能力.在温度为30℃、pH 7～8、盐度5～10%的条件下,H17均能高效降解苯酚,最高降解率可达到88.5%.该菌株降解苯酚动力学符合Halane模型,经拟合其生长参数为μ_(max)=0.31 h~(-1),K_S=191.63 mg·L~(-1),K_i=683.05 mg·L~(-1).研究显示H17具有在高盐环境下降解和耐受苯酚的能力,同时对环境有较强的适应性,体现出其在高盐含酚废水实际处理中具有良好的应用价值.     

Mn2+在黄杆菌FCN2菌株降解芘过程中的作用研究

利用紫外分光光度法和原子吸收分光光度法研究了Mn²⁺在菌株FCN2生长细胞、悬浮细胞、粗酶液降解芘时的影响作用.在菌体生长期加入0.1～5 mmol/L的Mn²⁺,对菌体的生长及生长量无影响,但对芘的降解均有促进作用,以加入0.1mmol/L的Mn²⁺效果最好,其对芘的平均降解率是对照的1.26倍.此时菌株富集的Mn²⁺为0.025mmol/L;在用无外加Mn²⁺培养的菌株FCN2悬浮细胞降解芘时,加入0.5 mmol/L的Mn²⁺,芘的平均降解率为对照的1.67倍.降解反应发生72h后菌株富集的Mn²⁺为0.18mmol/L;在酶促降解时加入0.1mmol/LMn²⁺,平均降解率为对照的1.30倍.结果表明,在不同时期加入的Mn²⁺对降解芘均有一定的促进作用.     

Degradation of polycyclic aromatic hydrocarbons by Pseudomonas sp. JM2 isolated from active sewage sludge of chemical plant

It is important to screen strains that can decompose polycyclic aromatic hydrocarbons (PAHs) completely and rapidly with good adaptability for bioremediation in a local area. A bacterial strain JM2, which uses phenanthrene as its sole carbon source, was isolated from the active sewage sludge from a chemical plant in Jilin, China and identified as Pseudomonas based on 16S rDNA gene sequence analysis. Although the optimal growth conditions were determined to be pH 6.0 and 37℃, JM2 showed a broad pH and temperature profile. At pH 4.5 and 9.3, JM2 could degrade more than 40% of fluorene and phenanthrene (50 mg/L each) within 4 days. In addition, when the temperature was as low as 4℃, JM2 could degrade up to 24% fluorene and 12% phenanthrene. This showed the potential for JM2 to be applied in bioremediation over winter or in cold regions. Moreover, a nutrient augmentation study showed that adding formate into media could promote PAH degradation, while the supplement of salicylate had an inhibitive effect. Furthermore, in a metabolic pathway study, salicylate, phthalic acid, and 9-fluorenone were detected during the degradation of fluorene or phenanthrene. In conclusion, Pseudomonas sp. JM2 is a high performance strain in the degradation of fluorene and phenanthrene under extreme pH and temperature conditions. It might be useful in the bioremediation of PAHs.     

鱼腥藻在满江红-鱼腥藻共生体系去除水中铀时的作用

通过分离有藻满江红中的鱼腥藻,获得无藻满江红和鱼腥藻.采用水培试验的方法分别研究了无藻满江红、有藻满江红和鱼腥藻对水中铀的去除行为,并采用红外光谱技术分析了它们富集铀前后化学基团的变化.结果表明,对于初始浓度分别为2.5和5.0 mg·L-1的铀溶液,在有藻满江红的作用下,分别经过27和36 d生物富集后浓度能降低至GB 23727-2009规定的0.05 mg·L-1排放标准以下,而在无藻满江红和鱼腥藻的作用下,铀浓度无法达到排放标准.相对于无藻满江红,鱼腥藻提高了有藻满江红对水中铀的富集量、富集系数及对铀的抗胁迫能力,从而提高了有藻满江红对水中铀的去除效率.红外分析表明,在富集铀后,与无藻满江红相比,有藻满江红体内出现了芳环和醇类或苷类化合物,而这些新的物质是鱼腥藻在富集铀的过程中产生的,其可能促进了有藻满江红对铀的富集.     

一株尼古丁高效降解菌株的分离鉴定和降解特性

从杭州利群环保纸业有限公司库存废烟叶中分离得到一株尼古丁高效降解菌株ZUTSKD.经形态、生理生化以及16S rDNA序列同源性分析,鉴定该菌株属于Pseudomonas sp.,命名为Pseudomonas sp. Strain ZUTSKD.该菌能以尼古丁为唯一碳氮源和能源生长,降解尼古丁最适温度为30℃,最适pH为7.5.添加氯化铵和葡萄糖有利于尼古丁的降解.在尼古丁的代谢产物中能够检测到3-(2,3,4-三氢-5-吡咯基)-吡啶,2,3′-二吡啶, 可天宁和3-羧基-吡啶.该研究表明,ZUTSKD菌株有很强的尼古丁降解能力,可用于处理烟草废弃物.     

Biodegradation of benzo[a]pyrene in soil by Mucor sp. SF06 and Bacillus sp. SB02 co-immobilized on vermiculite

Two indigenous microorganisms, Bacillus sp. SB02 and Mucor sp. SF06, capable of degrading polycyclic aromatic hydrocarbons （PAHs） were co-immobilized on vermiculite by physical adsorption and used to degrade benzo[a] pyrene （BaP）. The characteristics of BaP degradation by both free and co-immobilized microorganism were then investigated and compared. The removal rate using the immobilized bacterial-fungal mixed consortium was higher than that of the freely mobile mixed consortium. 95.3% of BaP was degraded using the co-immobilized system within 42 d, which was remarkably higher than the removal rate of that by the free strains. The optimal amount of inoculated co-immobilized system for BaP degradation was 2%. The immobilized bacterial-fungal mixed consortium also showed better water stability than the free strains. Kinetics of BaP biodegradation by co-immobilized SF06 and SB02 were also studied. The results demonstrated that BaP degradation could be well described by a zero-order reaction rate equation when the initial BaP concentration was in the range of 10--200 mg/kg. The scanning electronic microscope （SEM） analysis showed that the co-immobilized microstructure was suitable for the growth of SF06 and SB02. The mass transmission process of co-immobilized system in soil is discussed. The results demonstrate the potential for employing the bacterial-fungal mixed consortium, co-immobilized on vermiculite, for in situ bioremediation of BaP.     

异养脱氮菌株Bacillus sp. LY降解有毒有机污染物的研究

研究了异养脱氮菌株Bacillus sp. LY降解去除有毒有机污染物的性能.结果表明,菌株在同时实现有毒有机污染物去除与生物脱氮方面具有潜在优势.以典型有毒有机污染物苯酚、邻苯二酚、环境雌激素NP及其前驱物NPEOs为唯一碳源时,该菌株对4种物质的降解去除率分别达87%、77%、96%和80%,一级降解速率常数分别为0.191、0.112、0.435、0.147 d^-1.在分别以NP及NPEOs为唯一碳源的条件下,菌株对体系中的总氮亦有一定的去除,表现出异养脱氮能力.这些研究结果可为开发新型高效生物脱氮联合去除有毒有机污染物工艺打下理论基础.