酸性含硒含镉废水低温生物强化处理技术研究

本研究将枯草芽孢杆菌接种于活性污泥中,系统考察了酸性含硒含镉废水的低温生物强化处理效果.研究发现,活性污泥联合枯草芽孢杆菌对硒、镉的去除效率(Se:94.9%,Cd:99.1%)高于单一枯草芽孢杆菌(B菌)、混合枯草芽孢杆菌及活性污泥.影响因素试验结果表明,在20℃、pH 4.0、甲醇为碳源时生物强化硒、镉去除效率较高.利用SBR反应器处理酸性(pH 4.0)含硒(3.7～10.0 mg·L~(-1))含镉(5.0～11.2 mg·L~(-1))废水,在低温(8.0～10.8℃)条件下运行80个周期,硒、镉和COD的平均去除率分别为97.4%、90.7%和95.0%.红外光谱分析表明,酰胺基、芳香族C—H伸缩健在硒、镉去除中起到主要作用.透射电镜与能谱分析证实了生物强化污泥对硒、镉去除之后形成纳米颗粒物.通过高通量测序分析微生物群落结构,发现生物强化污泥中富集了Pseudomonas(假单胞菌属)、Dechloromonas(脱氯单胞菌)等典型硒还原菌.     

磺胺二甲基嘧啶(SM2)高效降解菌J2的分离筛选及降解特性研究

从城市污水处理厂活性污泥中分离得到一株能以磺胺二甲基嘧啶(SM2)为唯一碳源的菌株,经生理生化鉴定和16S rRNA基因序列同源性分析,将此菌鉴定为蜡样芽孢杆菌(Bacillus cereus),命名为J2.不同条件下的降解特性研究结果表明,J2菌株具有极高的SM2耐受性(100 mg·L~(-1))、较广的温度(20～30℃)和pH(6～8)适用范围;在温度30℃、pH=8.0、初始OD_(600)=0.1、SM2起始浓度为50 mg·L~(-1)的条件下,J2菌株在36 h内对SM2的降解率可达100%,降解效率远超目前已报道的其他SM2降解菌株,展现出了良好的应用潜力.J2菌株降解SM2过程中产生了5种主要中间代谢产物,分析推断其降解SM2的途径分为两条:①磺胺二甲基嘧啶分子首先在酶促反应作用下脱除SO_2,生成嘧啶环和苯胺环自由基,这两种自由基再经过环间耦合生成N-(4,6-二甲基嘧啶-2基)-1,4-二苯胺,该分子中的C—N键在活性氧物种的作用下断开生成苯胺和2-氨基-4,6-二甲基嘧啶;②在漆酶的作用下N~4键断裂,产生N-(3,5-二甲基嘧啶)-苯磺酰胺,之后N-(3,5-二甲基嘧啶)-苯磺酰胺的N—S键断裂,进一步形成2-氨基-4,6-二甲基嘧啶和苯亚砜.     

高效苯酚降解菌Bacillus sp.L5-1的分离及其降解特性

从污水处理厂活性污泥中分离筛选出一株高效苯酚降解菌L5-1,经菌落形态观察和16S rDNA基因测序,结果表明菌株L5-1为蜡样芽胞杆菌（Bacillus cereus）,美国国家生物信息中心（NCBI）的注册号为MN784421.将苯酚设置为唯一碳源,对其生长和苯酚降解特性展开研究.结果表明：菌株L5-1在10%接种量、温度30~35℃、pH值7~8的条件下,均能高效降解培养基中苯酚（培养基体积为100mL,初始苯酚浓度为500mg/L,14h时降解率>93%）.而在最优降解条件下（10%接种量,培养温度为35℃,pH值7.0,NaCl浓度为1%）,初始苯酚浓度为500mg/L,菌株在14h内的苯酚降解率可达97.1%;而当初始苯酚浓度为1000mg/L,菌株也可在46h内达到97.71%的降解率.运用Haldance方程动力学模拟菌株在不同浓度苯酚下的生长过程,其最大比生长速率为0.355h^-1,半饱合常数104.27mg/L,抑制常数为322.83mg/L,R²=0.997.菌株L5-1为目前已报道的Bacillus菌属中降解苯酚能力较强的菌株,为实际处理含酚废水中提供理论参考.     

Biosorption of Cu (II) ions by indigenous copper-resistant bacteria isolated from polluted coastal environment

The concentrations of copper in water and sediment samples of Cuddalore coastal area, Tamil Nadu, India were examined. The isolation, characterization, screening, and metal sorption capabilities of copper-resistant bacteria (CURB) also were investigated. Biosorption capabilities and growth time varied among the resistant isolates. All isolates reduced more than 80% of copper from copper-treated broth. However, Bacillus cereus CURB-4 strain absorbed more copper ions (99.9%) than other resistant isolates. Changes in structural group position and synthesis of extracellular polysaccharides (EPS) of CURB-4 strain under copper stress and controlled conditions also were examined. Under copper stress condition, CURB-4 produced more EPS (71 ± 0.6%) compared to control conditions (66 ± 0.4%). Fourier transform infrared spectroscopy revealed that the changes in functional group position of EPS were observed in stressed condition. Overall, synthesis of EPS in bacteria is a protective barrier against environmental stress.     

城市污泥中重金属镉对苏云金杆菌发酵的影响机制

为揭示苏云金杆菌（Bacillus thuringiensis,简称Bt）富集重金属的可行性,考察重金属对Bt发酵的影响,以生物毒性较大的重金属镉（Cd）为研究对象,以城市污泥为培养基,研究了添加Cd（II）对Bt生长和晶体蛋白合成的影响,分析了Bt发酵过程中Cd有效态质量浓度的变化,利用傅立叶红外光谱（FTIR）和X线光电子能谱（XPS）初步探讨了Cd（II）在Bt表面吸附的作用机制。结果表明：培养基中外加Cd（II）对Bt生长的影响主要取决于Cd有效态质量浓度,污泥培养基在支持Bt生长和降低重金属有效态质量浓度上均明显优于常规培养基;Bt对Cd（II）有较好的耐受性,当Cd有效态质量浓度在20 mg.L-1以下时,Bt能较好地完成生长代谢过程;光谱学分析表明Bt对Cd（II）有少量吸附,吸附位点主要为羟基,少量C=O基团也可与吸附反应,可有效降低重金属的生物有效性。     

蜡状芽孢杆菌不同菌株降解有机磷农药的特性分析

筛选分离降解微生物解决有机磷农药残留给水体和土壤环境带来的污染问题是一项可行的生物修复技术。采用富集培养和定时取样分析有机磷农药残留的方法,分离驯化出三株能够降解有机磷农药的细菌,研究了其形态特征和生理生化特性并对其16SrDNA序列进行了分析,同时比较了三菌株对甲基对硫磷（Methyl－parathion）、毒死蜱（Chlorpyrifos）和_二唑磷（Triazophos）的降解特性。结果表明：通过富集培养得到10菌株具有降解甲基对硫磷和毒死蜱的能力,比较确定HY-1、HY-2和HY-4三菌株作为研究对象,经鉴定为蜡状芽孢杆菌（Bacilluscereus）的不同菌株,三菌株在Genbank上的登录号分别为：eu915687、eu915686和eu915688。在甲基对硫磷质量浓度为50mg·L-1时,三菌株72h的降解率分别为91．7％、87．7％与92．4％．降解率无显著性差异（P〉0．05）,当甲基对硫磷质量浓度增加到100mg·L-1时,三菌株对甲基对硫磷的降解率有所下降,其中HY-2对甲基对硫磷的降解率下降最大达23％,且和其他两菌株有显著性差异（P〈0．05o三菌株72h对100mg·L-1毒死蜱的降解率分别达到64．8％、53．7％和56．5％,在不同的毒死蜱初始质量浓度下,HY-1和HY-4两菌株对毒死蜱的降解率无显著性差异（P〉0．05）,HY-2与HY-1、HY-4两菌株有显著性差异（P〈0．05o三菌株对三唑磷的降解率均较低,其中HY-2对初始质量浓度为100mg·L-1三唑磷的降解率最高仅为20．7％,其余两菌株对三唑磷的降解率比HY-2低且无显著性差异（P〉0．05o可以得出本研究分离得到的蜡状芽孢杆菌不同菌株对有机磷农药的降解存在多态性。     

一株产生物絮凝剂的Bacillus sp.的分离与鉴定

从重庆城南污水处理厂的活性污泥中分离筛选到一株絮凝剂产生菌,革兰氏染色呈阳性,好氧,有芽孢,可运动,短杆;分离菌的基因组DNA(G +C)mol%含量测定为5 0 .6 % ,16SrRNA序列分析表明,分离筛选出的絮凝剂产生菌的16SrRNA基因序列与Bacillus菌有高度同源性,从而确定絮凝剂产生菌RL - 2归于Bacillusbrevis菌属。絮凝实验表明:在有CaCl2 存在时用Bacillussp .RL - 2生物絮凝剂处理土壤悬液时用量少(10 0mL样品只需2mL絮凝剂) ,絮体形成快,矾花大,泥少;体系温度对活性影响很小,且在酸性与碱性条件下,絮凝活性均很高     

苏云金芽胞杆菌基因工程菌WG-001和1125BG在盆栽土壤中的安全性评估

自从遗传改造微生物诞生以来,其安全性问题受到了持续关注和研究.本文报道了苏云金芽胞杆菌基因工程菌WG-001和1125BG在盆栽大白菜的土壤环境中的安全性评估结果.实验结果表明:当用工程菌WG-001和1125BG分别以1013 cfu/cm2和1010 cfu/cm2的高土壤密度喷洒盆栽大白菜后,工程菌菌数迅速上升而后逐渐下降,最后分别以相对较低的菌数水平(105 cfu/g干土样和106 cfu/g干土样)在实验土壤中稳定存在;对土壤中土著细菌群落在有限时间(15d)内产生了一定影响,而对土著真菌与放线菌无显著影响;未发现工程菌WG-001中特异基因发生丢失;未检测到工程菌WG-001和1125BG中特异基因(WG-001为cry1Aa和cry1Ac,1125BG为gfp)向土著细菌、真菌、放线菌和盆栽大白菜的水平转移.本文实验结果显示,高密度喷洒的两种工程菌未显著影响实验土壤环境,具有很高的安全性.同时发现以不同的喷药量喷洒两种工程菌后,在盆栽土壤中其存活水平、菌数降低的速率以及对土著细菌的影响能力都表现出不同的特点.     

Identification of Bacillus methylotrophicus and its application in preventing fly maggot production in the environment北大核心CSCD

This research aimed to screen Bacillus spp. to prevent the production of fly maggot on kitchen wastes from soil in the experimental bases of the South China Agricultural University. A nutrient-rich medium was used to isolate the Bacillus spp. with high temperature treatment. The seventeen Bacillus strains were obtained and assigned to three groups by using Insertion Sequence based PCR (IS-PCR) DNA fingerprinting patterns. The homology of the 16S rDNA gene was 100% between Group I and Bacillus methylotrophicus CBMB 205T, 99.61% between Group II and Bacillus aerophilus 28KT, and 99.87% between Group III and Bacillus cereus ATCC 14579T. The results of kitchen waste tests proved that the representative bacteria of Group I RF2 had the ability to prevent fly maggot production on kitchen wastes. Furthermore, the results of physiological and biochemical tests, carbon utilization tests, and antagonists against plant pathogen tests showed that the bacteria from Group I RF2 had the ability to decompose glucose into pyruvate and then decarboxylate pyruvate to diacetyl under alkaline conditions, convert ammonia to intracellular amino acids or other kinds of nitrogenous compounds, use many kinds of carbon source for self-growth, and be antagonistic against plant pathogenic rice sheath blight disease, banana fusarium wilt, and Fusarium oxysporum. Besides, the bacteria from RF2 could secret organic acids to dissolve insoluble phosphate [Ca3(PO4)2] for its own growth by decreasing the pH. Group I RF2 was a strong plant-promotion bacterium, and had good prospects of application for preventing fly maggot production on kitchen wastes. © 2018 Science Press. All rights reserved.     

Degradation of chitin and antagonism in Phomopsis asparagi by Bacillus amyloliquefaciens MY001北大核心CSCD

About 10 billion tons of chitin, a natural polymer, is generated annually. However, the utilization of chitin was hindered because of the lack of a clean and effective degradation process. In this study, bacterial strains with chitosanase-producing ability were isolated from soil and identified using 16S rDNA, gyrA, and gyrB gene sequence analysis. In addition, the degradation of insoluble chitin and antifungal capability of the bacterial strains were investigated. One selected isolate was identified as Bacillus amyloliquefaciens strain MY001. It showed antagonistic effect on Phomopsis asparagi (Sacc.) Bubak and could degrade insoluble chitin. The combination of chitin/chitosan and MY001 strain could enhance the inhibition on the growth of P. asparagi. In conclusion, strain B. amyloliquefaciens MY001 can directly degrade chitin. The combination of MY001 strain with chitin shows more effective inhibition on P. asparagi growth. These results suggest the direct utilization of chitin against fungal diseases of crops. © 2018 Science Press. All rights reserved.