Removal of ammonium-N from ammonium-rich sewage using an immobilized Bacillus subtilis AYC bioreactor system

A self-design bioreactor system employing a fixed bed operation process with immobilized Bacillus subtilis AYC beads for NH4 +-N removal from slightly polluted water was proposed. Polyvinyl alcohol and Na-alginate were used as a gel matrix to entrap Bacillus subtilis AYC to form the immobilized beads. The NH4 +-N removal process was studied in a intermittent operation mode to examine the start-up and steady state behaviors of the immobilized AYC in the reactor. The results indicated that the reactor was in the start-up state during the first week. NH4 +-N began to be steadily removal since the second week, and the nitrogen removal rate was between 84.61% and 96.19% when the hydraulic retention time (HRT) was 30 min. To apply Bacillus subtilis AYC to develop a practical nitrogen removal system and further understand its nitrogen removal ability, the bioreactor was continuously operated under different experimental perameters. The results showed that under the optimum conditions of an HRT of 20 min and DO of 3.77–5.80 mg/L, the NH4 +-N removal rate reached 99.55%. The NH4 +-N removal rate increased as the C/N ratio increased. However, a high C/N may cause a high residual carbon level in the effluent, therefore, the most suitable C/N ratio was 10. In addition, the results showed that the bioreactor system could remove many types of nitrogen such as NH4 +-N, NO3??-N and organic-N, and had a good performance for inorganic nitrogen removal from sewage.     

Biological treatment of chicken feather waste for improved biogas production

A two-stage system was developed which combines the biological degradation of keratin-rich waste with the production of biogas. Chicken feather waste was treated biologically with a recombinant Bacillus megaterium strain showing keratinase activity prior to biogas production. Chopped, autoclaved chicken feathers (4%, W/V) were completely degraded, resulting in a yellowish fermentation broth with a level of 0.51 mg/mL soluble proteins after 8 days of cultivation of the recombinant strain. During the subsequent anaerobic batch digestion experiments, methane production of 0.35 Nm³/kg dry feathers (i.e., 0.4 Nm³/kg volatile solids of feathers), corresponding to 80% of the theoretical value on proteins, was achieved from the feather hydrolyzates, independently of the pre-hydrolysis time period of 1, 2 or 8 days. Cultivation with a native keratinase producing strain, Bacillus licheniformis resulted in only 0.25 mg/mL soluble proteins in the feather hydrolyzate, which then was digested achieving a maximum accumulated methane production of 0.31 Nm³/kg dry feathers. Feather hydrolyzates treated with the wild type B. megaterium produced 0.21 Nm³ CH₄/kg dry feathers as maximum yield.     

2株球形棕囊藻溶藻细菌的分离及鉴定

从珠海赤潮海水中分离出2株溶藻细菌Y01和Y04,对球形棕囊藻均有显著的溶藻效果.结果表明,溶藻细菌Y01和Y04均为革兰氏阳性菌,对球形棕囊藻的溶解作用时间为6 d.借助显微镜及扫描电镜等观察Y01和Y04溶藻过程,发现2株溶藻细菌的溶藻方式均为直接裂解藻细胞.溶藻细菌Y01和Y04的16S rDNA系列分析表明,溶...     

一株解淀粉芽胞杆菌的分离鉴定及水质净化效果的研究

采用富集培养及摇瓶实验,从污染河沟底质活性污泥中分离筛选到1株对亚硝态有较强转化能力的菌株HN.经形态特征、生理生化特性、16S rDNA基因序列分析、细胞脂肪酸组成及Biolog全自动微生物鉴定系统分析,鉴定菌株HN为解淀粉芽胞杆菌(Bacillus amyloliquefaciens),并在3种模拟水体环境中对菌株HN的水质净化效果进行了初步研究.结果表明,该菌株在试验第4d能将高浓度模拟污水(水样Ⅰ)、鲫鱼养殖废水(水样Ⅱ)、鲤鱼和对虾养殖废水(水样Ⅲ)中初始浓度分别为6.47、1.20和4.50mg·L-1的亚硝态氮去除100%、46%和100%,而对照的转化率分别为-10.4%、-800%和16.0%,同时对硝态氮也表现较强的去除能力且基本不累积氨氮.环境耐受性试验表明,该菌株在20～45℃、pH5.0～10.0、海盐浓度0.5%～3.0%的范围内都能正常生长.研究认为,解淀粉芽胞杆菌HN能显著降低水体中亚硝态氮和硝态氮的含量,且环境耐受力强,有望将其开发成高效的水产养殖水体净化微生态制剂.     

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).