巨大芽孢杆菌对土壤理化性质及植物富集镉锌的影响

不同生物之间存在复杂相互作用,构建多元生物协同修复有利于提高修复效果.微生物群落繁殖快,适应性强以及抗逆性等优势在土壤重金属污染治理中发挥着重要作用.以巨大芽孢杆菌（Bacillus megaterium）为供试菌株,以包心芥菜［Brassica juncea（L.） Czerniak.］为供试植物,探究二者联合作用下修复Cd和Zn污染土壤的可行性.首先开展60 d室内培养试验,明确巨大芽孢杆菌活化土壤Cd和Zn含量的潜力及改善土壤质量的可行性;进而通过盆栽试验探究巨大芽孢杆菌对包心芥菜富集重金属的影响.结果表明室内培养条件下,巨大芽孢杆菌能够显著降低土壤pH值,土壤有效态Cd和Zn含量显著增加,与对照相比,增幅分别为24%~47%和11%~13%;同时土壤磷酸酶（ALP）、蔗糖酶（SU）和脲酶（UR）活性明显改善.盆栽试验结果表明,与对照相比,接菌处理显著提高了包心芥菜生物量,增幅为10%~23%.同时植物富集Cd和Zn含量升高：植物地上和地下部分Cd富集浓度分别为对照组的1.61~1.70倍和1.05~1.15倍;不同部位对Zn的富集浓度分别为对照的1.38~1.61倍和1.47~1.53倍.相关性分析结果表明,土壤pH值是引起土壤重金属有效态活性和酶活性改变的关键因素.接菌处理下,植物过氧化氢酶（CAT）活性对增强植物抗逆性更为显著.试验结果初步证实了巨大芽孢杆菌-包心芥菜联合修复Cd和Zn污染土壤的可行性.     

微波无极紫外方法对二沉池出水的消毒研究

采用自制微波无极紫外连续式消毒反应器,对二沉池出水进行消毒.紫外光源为自制圆柱形无极灯,灯内充汞10mg、氩气压强666.61Pa紫外光强5.07mW/cm²,进水流量0.072L/s,出水细菌总数2CFU/mL,大肠菌群数0.微波无极紫外作用过程中,微波破坏细胞壁细胞膜结构使蛋白质、K⁺等胞内物质产生渗漏,这种不可逆的破坏过程有效抑制紫外光复活现象.     

Assessment of the in vivo genotoxicity of a new formulation of Bacillus thuringiensis var israelensis SH-14

Biological control agents have become a useful alternative for the reduction of the use of chemical insecticides. LABIOFAM (Cuba) is developing a new formulation of a biolarvicide that possesses as active biological agent Bacillus thuringiensis var israelensis serotype H14. In order to evaluate the genotoxicity of this new formulation, an in vivo battery test was used: micronucleus (MN), chromosome aberrations (CAs), and sperm morphology (SM) assays. A dose of 6.45?×?10⁸ spores was administered per animal via oral administration. Bone marrow cells were collected 24?h after a two day treatment for the MN assay, and 24?h after a unique treatment for the CA assay, using cyclophosphamide as the positive control. Sperm cells were collected at 5 weeks from the first of five administrations for the SM test, using acrylamide as positive control. Bacillus thuringiensis var israelensis serotype H14 failed to show either a significative increase of micronucleated polychromatic erythrocytes, chromosomal aberrations, or sperm abnormalities. Acute oral administration of a high dose of Bacillus thuringiensis var israelensis serotype H14 did not produce mutagenic effects in bone marrow or sperm cells.     

Protoplast preparation and regeneration conditions of Bacillus subtilis ZGL14 producing alkaline pectinase北大核心CSCD

The protoplast fusion technique is one of the important microbial breeding methods. The main factors that affect protoplast preparation and regeneration were investigated using Bacillus subtilis ZGL14 as the starter strain. The optimal conditions for protoplast preparation and regeneration were determined in this study by optimizing the cell age, lysozyme concentration, enzymolysis time, enzymolysis temperature, pH values of the osmotic pressure stabilizer, regeneration medium types, and culture type. The results indicated that the optimal condition for protoplast preparation of B. subtilis ZGL14 was as follows: 9 h of cell age, 0.2 mg/mL of lysozyme concentration, 10 min of enzymolysis time, 32 °C of enzymolysis temperature, and 7.0 of osmotic pressure stabilizer pH value. Under this condition, the protoplast formation and regeneration rates were > 98% and > 30%, respectively. A good protoplast regeneration effect can be achieved using the regeneration medium with sodium succinate as osmotic pressure stabilizer in combination with Ca2+. The single-layer culture was better than double-layer for regeneration. The optimal condition for protoplast preparation and regeneration of B. subtilis ZGL14 was determined in this study, which provides technical support for protoplast fusion breeding of B. subtilis. © 2018 Science Press. All rights reserved.     

Isolation,identification, and the degradation characteristics of a thiobencarb-degrading bacterium Bacillus sp. T2北大核心CSCD

Thiobencarb, a thiocarbamate herbicide, is widely used to control weeds in rice paddies. Screening for highly efficient thiobencarb-degrading bacteria is important for the bioremediation of thiobencarb-contaminated environments. The aim of this study was to isolate and identify highly efficient thiobencarb-degrading bacteria and to identify the degradation pathway and the degrading properties. The thiobencarb-degrading strain was isolated using methods of microbiological acclimation and enrichment and was then identified using a 16S rRNA phylogenetic analysis. The degrading properties of the isolated bacterium were determined by single-factor experiments, and the degradation products were identified using gas chromatography-mass spectrometry (GC-MS). A thiobencarb-degrading strain T2, which can utilize thiobencarb as the sole source of carbon for energy and growth, was isolated from paddy soil. Strain T2 degraded more than 98.3% of 0.4 mmol/L of thiobencarb within 36 h. It was preliminarily identified as Bacillus sp. T2 according to the 16S rRNA gene analysis and from its morphological, physiological, and biochemical characteristics. The metabolic products of the thiobencarb degradation for strain T2 were identified as 4-chlorobenzyl mercaptan, 4-chlorobenzaldehyde, and 4-chlorobenzoic acid by the GC-MS. Based on metabolite identification, it was speculated that thiobencarb degradation in strain T2 was initiated by the hydrolysis of the thioester bond to produce 4-chlorobenzyl mercaptan, which was further oxidized to 4-chlorobenzaldehyde and 4-chlorobenzoic acid. The thiobencarb degradation that was initiated by the hydrolysis of the thioester bond by strain T2 is a new metabolic pathway, which provides valuable research material and reliable experimental data for revealing the metabolic process and mechanism of thiobencarb microbial degradation in soil. The strain Bacillus sp. T2 has a very high degradation efficiency, suggesting it is a good prospect for microbial remediation in thiobencarb-polluted environments. © 2018 Science Press. All rights reserved.     

一株溶藻细菌对铜绿微囊藻生长的影响及其鉴定

从山东黄岛边某富营养化池塘中分离得到1株具有溶藻作用的菌株(J1),研究了其对铜绿微囊藻(Microcystis aeruginosa)的抑制效果、作用方式以及细菌培养基、菌株与铜绿微囊藻不同生长阶段等因素对溶藻效果的影响,并对该菌株进行了生理生化鉴定.结果表明,将牛肉膏蛋白胨培养基加入藻液,培养基与藻液的投放体积比为6%时,9d内对藻的生长无影响.将初始浓度为6×107 cfu/mL的菌液加入藻液,共培养第9d,铜绿微囊藻的去除率达87%以上.对数期的J1细菌具有较好的溶藻效果,作用于稳定期铜绿微囊藻的实验组,藻的去除率较低.J1通过分泌溶藻物质的间接作用方式抑制铜绿微囊藻的生长,且溶藻活性物质具有一定的热稳定性.根据生理生化及16S rDNA序列分析鉴定,菌株J1属于芽胞杆菌属(Bacillus).     

红树林沉积物Bt菌株的分离与cry基因型的鉴定（Isolation of Bacillus thuringiensis Strains from Mangrove Sediment Samples and Identification for cry Genes）

从收集自泉州洛江红树林保护区的400个沉积物土壤样品中,分离到18株苏云金芽孢杆菌(Bacillus thuringiensis,Bt).利用PCR-RFLP体系对Bt菌株杀虫晶体蛋白的基因型进行了鉴定,发现15株Bt含有cry1基因,4株含有cry2基因,有3株分别含有cry7、cry8、cry9基因.克隆了一个新型的cry2Ab基因,其编码蛋白与现有Cry2Ab型杀虫晶体蛋白的最高同源性为95%.     

苏云金芽孢杆菌生物杀虫剂发酵生产的影响因素及其工艺选择

化学杀虫剂的长期使用给生态环境造成了严重破坏,也使害虫种群的抗药性日益提高,生物杀虫剂以其＂绿色环保＂的特点引起人们的广泛关注。其中,苏云金芽孢杆菌（Bacillus thuringiensis）制剂是目前世界上产量最大、应用最广的生物杀虫剂。它对鳞翅目、双翅目、鞘翅目、螨类等许多有害昆虫有毒杀作用,而对人类、动物和农作物无害。长期以来,人们一直致力于苏云金芽孢杆菌发酵过程的研究,以期获得高毒效的生物杀虫剂产品。本文首先对苏云金芽孢杆菌发酵生产的各种影响因素进行了综合分析,将影响因素分为培养条件和培养基组分两类,得出最佳培养条件为温度：（30±1）℃,pH：7.0±0.1,搅拌速度：400～600r·min-1,通气量：1∶0.6～1.2（发酵培养基体积与每分钟通入空气的体积之比）,接种时间：对数期初;最佳培养基配比为碳氮比：8～10∶1,无机盐含量：KH2PO4或K2HPO4为0.075%～0.2%;MgSO4·7H2O为0.075%～0.3%;CaCO3为0.075%～0.15%;MnSO4·H2O、FeSO4·7H2O各为0.002%。其次,对当前研究与工业化生产中的各种发酵工艺进行了评述,总结了现有发酵工艺的优缺点。在现有研究基础上,降低培养基原料成本、改进发酵工艺和采用基因学手段构建高效工程菌株将成为未来研究热点。     

Bio-remediation of acephate–Pb(II) compound contaminants by Bacillus subtilis FZUL-33

Removal of Pb~(2+)and biodegradation of organophosphorus have been both widely investigated respectively. However, bio-remediation of both Pb~(2+)and organophosphorus still remains largely unexplored. Bacillus subtilis FZUL-33, which was isolated from the sediment of a lake, possesses the capability for both biomineralization of Pb~(2+)and biodegradation of acephate. In the present study, both Pb~(2+)and acephate were simultaneously removed via biodegradation and biomineralization in aqueous solutions.Batch experiments were conducted to study the influence of p H, interaction time and Pb~(2+)concentration on the process of removal of Pb2+. At the temperature of 25°C, the maximum removal of Pb~(2+)by B. subtilis FZUL-33 was 381.31 ± 11.46 mg/g under the conditions of p H 5.5, initial Pb~(2+)concentration of 1300 mg/L, and contact time of 10 min. Batch experiments were conducted to study the influence of acephate on removal of Pb~(2+)and the influence of Pb2+on biodegradation of acephate by B. subtilis FZUL-33. In the mixed system of acephate–Pb2+, the results show that biodegradation of acephate by B. subtilis FZUL-33 released PO43+, which promotes mineralization of Pb2+. The process of biodegradation of acephate was affected slightly when the concentration of Pb2+was below 100 mg/L. Based on the results, it can be inferred that the B. subtilis FZUL-33 plays a significant role in bio-remediation of organophosphorus-heavy metal compound contamination.     

碱性β-聚糖酶产生菌选育及产酶条件优化

从碱性土样中分离到数十株产碱性β聚糖酶类的细菌,经摇瓶反复筛选后,得到一株碱性β聚糖酶产量较高的耐碱性细菌．经初步鉴定,属短小芽孢杆菌（Ｂａｃｉｌｕｓｐｕｍｉｌｕｓ）．其纤维素酶作用的最适ｐＨ为７．６,最适θ为６０℃,木聚糖酶的作用最适ｐＨ为９．０,最适θ为５５℃．该菌的最适生长ｐＨ为８．０,最适产酶θ为２８～３２℃．木聚糖与山梨糖分别是木聚糖酶和纤维素酶的良好诱导物．以麸皮为碳源,产酶的最适浓度为５％．添加尿素和（ＮＨ４）２ＳＯ４为氮源可提高纤维素酶酶活２倍,木聚糖酶酶活１倍．发酵周期为６０ｈ,纤维素酶酶活最高可达１．２１ＩＵｍＬ－１,木聚糖酶酶活可达４３ＩＵｍＬ－１．