Phylogenetic relationships among Trichodinidae (Ciliophora : Peritricha) derived from the characteristic values of denticles

The phylogenetic relationships among trichodinids remain obscure. As an important diagnostic marker, the morphology of the denticles in the adhesive disc as well as the adoral spiral has been widely used in generic discrimination and species identification of trichodinids. We studied the characters of denticles of the ten genera of Trichodinidae and the sole genus Urceolaria of Urceolariidae by using a quantitative method. The characteristic values were used to generate Manhattan distance, on which the dendrogram was based to construct with the Unweighted Paired Group Method using the Arithmetic mean (UPGMA). The investigations show that all the genera of the family Trichodinidae were clearly separate from the outgroup Urceolaria, and within the Trichodinidae: (i) Dipartiella grouped with Trichodinella and Tripartiella and lay in the closest position to the outgroup with a low dissimilarity, suggesting Dipartiella might be the most primitive genus in the family; (ii) Hemitrichodina clustered in a single clad and lay in the farthest position to the outgroup with the highest dissimilarity, indicating that it might be the most advanced genus; and (iii) the other 6 genera, Trichodina, Paratrichodina, Semitrichodina, Vauchomia, Pallitrichodina and Trichodoxa clustered in a big clad with very low dissimilarity, showing that they are closely related to each other. We discuss the evolutionary trend of the denticle and conclude that the denticles of the adhesive disc should be an apomorphic feature of the trichodinids and their changes could reflect the evolutionary tendencies of these ciliates.

Effect of replacement of dietary fish meal by meat and bone meal and poultry by-product meal on growth and feed utilization of gibel carp, Carassius auratus gibelio

Triplicate groups of gibel carp Carassius auratus gibelio (initial body weight: 5.25 +/- 0.02 g) were fed for 8 weeks at 20-25 degreesC on five isonitrogenous (crude protein: 400 g kg(-1)) and isoenergetic diets (gross energy: 17 kJ g(-1)). Meat and bone meal (MBM) or poultry by-product meal (PBM) were used to replace fish meal at different levels of protein. The control diet contained fish meal as the sole protein source. In the other four diets, 150 or 500 g kg(-1) of fish meal protein was substituted by MBM (MBM15, MBM50) or PBM (PBM15, PBM50). The results showed that feeding rate for the MBM50 group was significantly higher than for other groups except the PBM50 group (P < 0.05). Growth rate in the MBM15 group was significantly higher than that in the control (P < 0.05), while there was no significant difference in growth between the control and other groups (P > 0.05). Feed efficiency and protein efficiency ratio in MBM50 was significantly lower while that in MBM15 was significantly higher (P < 0.05). Replacement of fish meal by MBM at 500 g kg(-1) protein significantly decreased apparent dry matter digestibility (ADC(D)) and gross energy (ADC(E)) while apparent protein digestibility (ADC(P)) was significantly decreased by the replacement of MBM or PBM (P < 0.05). The results suggest that MBM and PBM could replace up to 500 g kg(-1) of fish meal protein in diets for gibel carp without negative effects on growth while 150 g kg(-1) replacement by MBM protein improved feed utilization.

Effect of replacement of fish meal by meat and bone meal and poultry by-product meal in diets on the growth and immune response of Macrobrachium nipponense

The potential use of poultry by-product meal (PBM) and meat and bone meal (MBM) as alternative dietary protein sources for juvenile Macrobrachium nipponense was studied by a 70-day growth trial. Triplicate groups of M. nipponense (initial body weight: 0.37 g) were fed at 20.7-22.4 degreesC on each of the five isoenergetic and isonitrogenous diets (protein content about 38%) with different replacement of fish meal by MBM or PBM. The control diet used white fish meal as the sole protein source, the other four diets were prepared with 15% or 50% fish meal protein substituted by either MBM (MBM15, MBM50) or PBM (PBM15, PBM50). The results showed that replacement of fish meal by MBM in diets did not affect growth performance of M. nipponense (P > 0.05), while specific growth rate in PBM15 was significantly higher than that in other groups (P < 0.05). Survival rates of shrimp fed with MBM15 diet were significantly higher than that in other groups (P < 0.05). No significant differences in immunological parameters, including total haemocyte count (THC), phenoloxidase activity (PO) and respiratory burst (O-2(-)), were observed between the shrimps that were fed five experimental diets, and all determined immunological parameters in control groups were slightly higher than those in replacement groups. In conclusion, either MBM or PBM investigated could replace up to 50% fish meal protein in diets for M. nipponense. (C) 2003 Elsevier Ltd. All rights reserved.

Determination of phthalate acid esters plasticizers in plastic by ultrasonic solvent extraction combined with solid-phase microextraction using calix[4]arene fiber

Ultrasonic solvent extraction combined with solid-phase microextraction (SPME) with calix[4]arene/hydroxy-terminated silicone (C[4]/OHTSO) oil coated fiber was used to extract phthalate acid esters (PAEs) plasticizers in plastic, such as blood bags, transfusion tubing, food packaging bag, and mineral water bottle for analysis by gas chromatography (GC). Both extraction parameters (i.e. extraction time, extraction temperature, ionic strength) and conditions of the thermal desorption in a GC injector were optimized by analysis of eight phthalates. The fiber shows wonderful sensitivity and selectivity to the tested compounds. Owing to its high thermal stability (380 degreesC), the carryover effect that often encountered when using conventional fibers can be reduced by appropriately enhancing the injector temperature. The method showed linear response over two to four orders of magnitude with correlation coefficients (r) better than 0.996, and limits of detection (LOD) ranged between 0.006 and 0.084 mug l(-1). The relative standard deviation values obtained were less than or equal to 10%. bis-2-Ethylhexyl phthalate (DEHP) was the sole analyte detected in these plastics and recoveries were in the ranges 95.5-101.4% in all the samples. (C) 2004 Elsevier B.V. All rights reserved.

Isolation and Characterization of a Methomyl-Degrading Paracoccus sp mdw-1

Methomyl, an extremely toxic pesticide, is widely used in agriculture. A strain named mdw-1 capable of degrading methomyl rapidly was successfully isolated from activated sludge in this study. It could utilize methomyl as the sole carbon or nitrogen source. The optimal temperature and medium pH for its growth and methomyl biodegradation were 30 degrees C and 7.0, respectively. It was identified as a Paracoccus sp. according to its morphological features, physiological and biochemical characteristics, and phylogenetic analysis based on the sequence of 16S rDNA. Gas chromatography-mass spectrometry (GC-MS) analysis showed that methomyl could be completely transformed to S-methyl-N-hydroxythioacetamidate in 10 h of incubation with the isolate mdw-1.

青稞B组醇溶蛋白基因与上游调控区的克隆及基因表达

高等植物种子胚乳贮藏蛋白是种子发芽时的主要氮源，也是人类和动物食用植物蛋白的主要来源。大麦种子胚乳贮藏蛋白主要是醇溶蛋白（hordeins），占大麦胚乳总蛋白的50–60%。根据大麦醇溶蛋白的大小和组成特点，大麦醇溶蛋白被划分为三种类型：富硫蛋白亚类(B，γ-hordeins)、贫硫蛋白亚类（C-hordeins)以及高分子量蛋白亚类(D-hordeins)。B组和C组醇溶蛋白是大麦胚乳的两类主要贮藏蛋白，它们分别占大麦总醇溶蛋白成分的70–80%和10–12%。遗传分析表明，大麦B、C、D和γ-组醇溶蛋白分别是由位于大麦第五染色体1H（5）上的Hor2、Hor1、Hor3和Hor5位点编码。Hor2位点编码大量分子量相同但组成不同的B组醇溶蛋白（B-hordein）。B-hordein的种类、数量和分布是影响大麦酿造、食用及饲养品质的重要因素之一。为深入了解B-hordein基因家族的结构和染色体组织，探明Hor2位点基因表达的发育调控机制，最终达到改良禾谷类作物籽粒品质的目的，本研究以青藏高原青稞为材料，采用同源克隆法，分别克隆B-hordein基因和启动子，通过原核生物表达验证B-hordein基因功能，并利用实时定量PCR探索B-hordein基因表达时空关系，取得如下研究结果： 1. 以具有特殊B组醇溶蛋白亚基组成的9份青藏高原青稞为材料，根据GenBank中三个B-hordein基因序列（GenBank No. X03103, X53690和X53691）设计一对引物，通过PCR扩增，获得23个B-hordein基因克隆并对其进行了序列分析。核苷酸序列分析表明，所有克隆均包含完整的开放阅读框。有11个克隆都存在一个框内终止密码子，推测这11个克隆可能是假基因。推测的氨基酸序列分析表明，所有大麦B-hordein具有相似的蛋白质基本结构，均包括一个高度保守的信号肽、中间重复区以及C-端结构域。不同大麦种重复区内重复基元的数目有较大差异。青稞材料Z07–2和Z26的B-hordeins仅具有12个重复基元结构，更接近于野生大麦。这些重复基元数目的差异导致了重复区序列长度和结构的变异。这种现象极可能是由于醇溶谷蛋白基因在进化过程中染色体的不平衡交换或复制滑动所造成的。对所克隆基因和禾本科代表性醇溶谷蛋白基因进行聚类分析，结果表明所有来自栽培大麦的B-hordeins聚类成一个亚家族，来自野生大麦的B-hordeins以及普通小麦的LMW-GS聚类成另外一个亚家族，表明这两个亚家族的成员存在显著差异。此外，我们发现B-hordein基因推测的C-末端序列具有一些有规律的特征：即具有相同C-末端序列的B-hordein基因在系统发生树中聚类为同一个亚组（除BXQ053，BZ09-1，BZ26-5分别单独聚为一类外）。这个特征将有助于我们对所有B组醇溶蛋白基因家族成员进行分类，避免了在SDS-PAGE电泳图谱上仅依靠大小分类的局限性。 2. 根据上述克隆的青稞B-hordein基因的5’端序列设计三条基因特异的反向引物，以青稞Z09和Z26的基因组DNA为模板，采用SON-PCR和TAIL-PCR技术分离克隆出8个B-hordein基因的上游调控序列（命名为Z09P和Z26P）。序列分析表明，推测的TATA box位于–80 bp，CAAT–like box位于–140 bp处。此外，Z09P和Z26P中有六个序列在–300 bp处均存在一个由高度保守的EM基序和类GCN4基序构成的胚乳盒(Endosperm Box，EB)，在约–560 bp处存在一个胚乳盒类似结构。而Z09P-2和Z26P-3不存在保守的胚乳盒或其类似结构，预示着这两个启动子所调控的基因表达可能受不同类型反式作用因子的调节，推测该启动子对基因的表达调控具有多样性。 3. 将B-hordein基因的开放阅读框定向克隆到表达载体pET-30a中，将其导入大肠杆菌表达菌株BL21中进行外源基因的诱导表达以验证所克隆基因的功能。结果表明仅含重组子pET-BZ07-2和pET-BZ26-5的BL21细菌有目的表达蛋白产生。在诱导3 h时的蛋白表达量最高；3 mM IPTG诱导的蛋白表达量要高于1 mM IPTG诱导的表达量。这为分离纯化B-hordein蛋白以及进一步研究其对大麦籽粒品质的影响奠定基础。 4. 根据从青稞Z09和Z26中分离克隆的B-hordein基因序列设计一对基因特异的引物，同时，选择大麦α-微管蛋白基因（GenBank no. U40042）为看家基因并设计特异引物，利用实时荧光定量PCR检测了青稞籽粒4个胚乳发育时间段的B-hordein基因表达，荧光定量结果显示：两份材料中B-hordein基因的表达量均随发育过程的进行而逐渐升高。Z09中B-hordein基因在开花后7天开始转录，而Z26开花4天后就有低水平B-hordein的表达，这表明Z26中B-hordein基因可能比Z09表达的较早或者Z09中B-hordein基因表达水平较低以致于不能被检测到。此外，在4个不同的胚乳发育时期中，Z26中B-hordein基因的表达量均高于Z09材料。在开花12天到18天的过程中，Z09和Z26中B-hordein基因的表达水平有一个急剧性的升高。这说明在不同胚乳发育时期，Hor2位点的B-hordein等位基因变异体存在mRNA的差异表达。 Seed endosperm storage proteins in higher plants are the main resources of nitrogen for germinating and plant proteins for human and animals. Barley prolamins (also called hordeins) are the major storage proteins in the endosperm and account for 50–60% of total proteins. Hordeins are classically divided into three groups: sulphur-rich (B, γ-hordeins), sulphur-poor (C-hordeins) and high molecular weight (HMW, D-hordeins) hordeins based on the size and composition. B-hordeins and C-hordeins are two major groups and each respectively account for about 70-80% and 10-12% of the total hordein fraction in barley endosperm. Genetic analysis showed that B-, C-, C-, γ-hordeins are encoded by Hor2, Hor1, Hor3 and Hor5 locus on the chromosome 1H (5). Hor2 locus is rich in alleles that encode numerous heterogeneous B-hordein polypeptides. It is reported that B-hordein species, quantity and distribution are significant factors affecting malting, food and feed quality of barley. To understand comprehensively the structure and organization of B-hordein gene family in hull-less barley and explore the developmental control mechanisms of Hor2 locus gene expression and eventually to better exploitation in crop grain quality improvement, we isolated and cloned B-hordein genes and promotors of hull-less barley from Qinghai-Tibet Plateau by PCR, and testified their expression founction in bacteria expression system and explore their spatial and temporal expression pattern by quantitative real time PCR. Our results are as followed, 1. Twenty-three copies of B-hordein gene were cloned from nine hull-less barley cultivars of Qinghai-Tibet Plateau with special B-hordein subunits and molecularly characterized by PCR, based on three B-hordein genes published previously (GenBank No. X03103, X53690 and X53691). DNA sequences analyses confirmed that the six clones all contained a full-length coding region of the barley B-hordein genes. Eleven clones all contain an in-frame stop codon and they are probably pseudogenes. The analysis of deduced amino acid sequences of the genes shows that they have similar structures including signal peptide domain, central repetitive domain, and C-terminal domain. The number of the repeats was largerly variable and resulted in polypeptides in different sizes or structures among the genes. Twelve such repeated motifs were found in Z07–2 and Z26, and they are close to those of the wild barleys, and it is most probably caused by unequal crossing-over and/or slippage during replication as suggested for the evolution of other prolamins. The relatedness of prolamin genes of barley and wheat was assessed in the phylogenetic tree based on their polypeptides comparison. Our phylogenetic analysis suggested that the predicted B-hordeins of cultivated barley formed a subfamily, while the B-hordeins of wild barleys and the two most similar sequences of LMW-GS of T. aestivum formed another subfamily. This result indicated that the members of the two subfamilys have a distinctive difference. In addition, we found the B-hordeins with identical C-terminal end sequences were clustered into a same subgroup (except BXQ053，BZ09-1 and BZ26-5 as a sole group, respectively), so we believe that B-hordein gene subfamilies possibly can be classified on the basis of the conserved C-terminal end sequences of predicted polypeptide and without the limit of SDS-PAGE protein banding patterns. 2. The specific primers were designed according to the published sequences of barley B-hordein genes from Z09 and Z26. Using total DNA isolated from them as the templates, eight clones (designated Z09Pand Z26P) of upstream sequences of the known B-hordein genes was obtained by TAIL-PCR and SON-PCR. Sequences analysis shows that the putative TATA box was present at position –80 bp and CAAT-like box at position –140 bp. Besides, a putative Endosperm Box including an Endosperm Motif (EM) and a GCN4-Like Motif was found at position –300 bp in six clones, and another Endosperm-like box was found at positon –560 bp. While the Endosperm Box or Endosperm-like box was not found in Z09P-2 and Z26P-3. This may indicate that gene expression drived by the two promtors was probably controlled by different trans-acting factors and the genetic control mechanism of corresponding gene expression may be diverse. 3. The B-hordein genic region coding for the mature peptide was cloned into expression vector pET-30a and transformed into bacterial strain BL21 for identifying gene expression fountion. Protein SDS–PAGE analysis showed that only the transformed lysate with the pET-BZ07-2 and pET-BZ26-5 constructs produced proteins related to B-group hordeins of barley, and the mounts of proteins induced by 3 mM IPTG and 3 h were higher than other conditions. This established a base for isolating and putifying B-hordein and further exploring their effects on barley grain quality. 4. The gene-specific primers of B-hordein genes from Z09 and Z26 were used for the quantification of B-hordein gene expression. The α-tubulin gene from Hordeum vulgare subsp. vulgare (GenBank accession number U40042) was used as a control gene. The result shows the transcription of the B-hordein genes in Z09 was found 7 days after flowering, while the transcription of the B-hordein genes in Z26 was found 4 days after flowering, but at a very low level, and it suggested that the B-hordein genes in Z26 probably expressed earlier than those in Z09, or the B-hordein genes in Z09 expressed at so a lower level than Z26 that it can not detected. In addition, B-hordein genes in Z26 accession showed higher expression levels than those in Z09 in four developing stages. Furthermore, a progressive increase in the expression levels of the B-hordein genes between 12 and 18 days after anthesis was observed in both Z09 and Z26. It implies that the B-hordein allelic variants encoded by Hor2 locus exist the differential expression in mRNA levels of during barley endosperm development.

生物催化的Baeyer-Villiger氧化反应研究

Baeyer-Villiger氧化反应是一种很重要的化学反应，产生的许多中间体或产物可以被用来生产多种化学产品和药物。此反应具有多功能性，可以氧化多种羰基化合物，但是化学方法中的必需反应物——氧化剂在生产、储存、运输、反应的过程中都存在很多的不安全因素，反应的立体选择性也不强，而生物转化则具有底物选择性、立构选择性、化学选择性、对映选择性等一般化学反应中不具备的优点，在精细化工中占有很大的优势。在工业生物催化中有很好的应用前景。 为了研究生物催化的Baeyer-Villiger反应，我们从本实验室保藏菌种中分离筛选出一株能够以环己酮作为唯一碳源的菌株，进行初步研究并对其产物进行GC/MS定性，探讨了pH，装液量，底物浓度，培养时间，温度以及转速等条件对细菌生长的影响，并进一步研究了细菌的底物广谱性。 此菌株经鉴定属于邻单胞菌属Plesiomonas sp.)， 根据正交试验，确定了菌的最佳生长条件：底物浓度为1mL/L，底物浓度过高对菌株生长有抑制作用，转速为150 rpm ，温度为30℃ ，pH为7.0； 此菌株转化环己酮的产物通过GC/MS检测含有内酯，表明此菌株能够催化Baeyer-Villiger氧化反应；此菌株还能够以与环己酮有相似结构的环己烷，环戊酮等作为唯一碳源生长，说明此菌株底物利用范围比较广，用途比较广泛。 Baeyer-Villiger oxidation is an important chemical conversion, its products and intermediates can be used to produce a lot of medicine and fine chemicals. Its success is largely due to its versatility: a variety of carbonyl compounds can be oxidized, a large number of functional groups are tolerated, the regiochemistry is highly predictable and so on, but the oxidants that the traditional chemistry way needs have a number of problem in their production, storage, transportation and reaction, Chemistry way has not a high stereochemistry yet. However, biotransformations have many attractive characters, such as substrate-, stereo-, chemo- and enantioselectivity, so it has a great advantage in the fine chemical industry and has a bright prospect in the industrial biological catalysis. In order to study Baeyer-Villiger oxidation, we isolated a strain which can utilize cyclohexanone as sole carbon source and had a primary research on it. Its product was identified by GC/MS. Effects of pH, volume, concentration of cyclohexanone, cultivating time, temperature and rotate speed on the growth of bacteria were discussed, and the other organic substrates were also studied. The strain was identified as Plesiomonas sp.. The result of orthogonal test made it sure that the best growth condition of the strain is: rotate speed 150 rpm, temperature 30℃, pH7.0, concentration of cyclohexanone1ml/L. There is caprolactone in the product of the fermentation with cyclohexanone as substrate by GC/MS,which indicated that the strain can catalyse Baeyer-Villiger oxidation.In addition,the strain can utilize other organic substrates having the similar structure with cyclohexanone such as cyclohexane, cyclopentanone, Swertiamarin as sole carbon source.So the strain can be applied extentively.

超声与磁粉强化含单宁废水生物处理研究

单宁是一种典型的有毒难降解污染物，在制革、造纸、制药、印染等行业废水中广泛存在，对水环境造成污染并且影响废水生物处理效果。本研究针对含单宁废水生物处理效率低、较高浓度时微生物受抑制且污泥容易膨胀等问题，采用超声和磁粉来强化含单宁废水生物处理，研究超声和磁粉对微生物活性、污染物去除及污泥沉降性能的影响，并对其作用机理进行了分析和探讨。 研究结果表明，活性污泥系统中单宁酸容积负荷可以达到1.8kgCOD/(m3·d)，单宁酸和COD去除率分别达到85.2%和79.6%，但如果负荷进一步增大则微生物活性迅速降低。系统在pH 5~8、温度20~35℃、DO>1 mg/L的条件下具有较好的单宁酸降解效果和处理稳定性。单宁降解动力学参数为：μmax =0.208h-1；Ks=226mg/L；Ki=522mg/L；kd=0.0092h-1；Y =0.594。 磁粉对系统处理效果和污泥沉降性能有一定的促进作用，且效果要优于外磁场。适宜的磁粉粒径和投加量分别为0.05~0.15mm和1.0g/L，COD去除率比对照系统提高6.4%，SVI降低28.6%，污泥絮体结构紧密。磁粉强化主要是通过其对污泥菌胶团的凝聚、吸附作用以及对微生物活性的强化作用实现。 在适当强度（0.4W/cm2）和辐照时间（20min）的超声作用下污泥絮体和细胞膜通透性增大，酶分泌也增多，系统的COD去除率比对照提高了8.8%，单宁酶酶活提高了11%。但超声也使污泥絮体结构松散，沉降性能下降，SVI比对照系统升高9.3%。 由于污泥流失加剧导致污泥浓度相对较低，声磁联合强化系统相对于磁粉强化系统其处理效果并没有提高。但相对于单纯活性污泥系统，声磁联合作用下系统处理效果、污泥沉降性能以及系统运行稳定性都得到明显改善。本研究为难降解废水的生物处理提供了一个新的思路。 Tannins are typical refractory and toxic pollutants that commonly exist in wastewater from dye, medicine, paper and leather industries and cause many problems associated with environmental pollution and biological treatment of wastewater. Biological treatment efficiency of tannin-containing wastewater is usually low owing to its biological toxicity and low biodegradability, microbes are usually inhibited under high tannin concentration and sludge bulking frequently occurs. In this study, ultrasound and magnetic powder were used to improve the biological treatment performance of simulated tannic acid-containing wastewater. The effects of ultrasonic irradiation and magnetic powder on microbial activity, tannic acid degradation rate and sludge sedimentation were investigated. The augmentation mechanisms were analyzed and discussed. The experimental results showed that the microbes were prominently inhibited under high tannic acid concentration, but moderate degradation efficiency can be maintained under a tannic acid load of up to 1.8kgCOD/(m3·d), with the tannic acid degradation and COD removal percentage of 85.2% and 79.6% respectively. The highest degradation rates and treatment stability were achieved at pH range of 5~8, temperature range of 20~35℃ and DO concentration of above 1mg/L. The kinetic parameters were estimated, including: μmax =0.208h-1；Ks=226mg/L；Ki=522mg/L；kd=0.0092h-1；Y =0.594. The microbial activity, tannic acid degradation rate and sludge sedimentation were improved by adding Fe3O4 magnetic powder, and the augmentation performance was better than external magnetic field. The appropriate particle size and dosage of magnetic powder were found to be 0.05~0.15mm and 1.0g/L, respectively, under which the COD removal percentage was improved by 6.4% and SVI value decreased by 28.6%, and compact floc structure was observed. This was mainly caused by the flocculation and adsorption effects of magnetic powder against sludge floc and the stimulation of microbial activity under appropriate magnetic field. Under appropriate ultrasonic irradiation (ultrasonic intensity 0.4W/cm2, ultrasonic irradiation time 20min), the permeability of floc and cell membrane are improved, transfer of substrate and oxygen were reinforced; meanwhile, more enzyme were produced by microbes under the slight damage caused by ultrasound. However, the floc structure became loose under ultrasonic irradiation, leading to relatively poor sedimentation, with the SVI value 9.3% higher than the control system. Although the magnetic powder-ultrasonic irradiation combined augmentation system showed no improvement in treatment performance compared with sole magnetic augmentation system owing to its relatively low sludge concentration, it guaranteed the stable operation of system, meanwhile the tannic acid degradation and sludge sedimentation were significantly improved compared with sole activated sludge system. This study gives a new idea for biological treatment of refractory wastewater.

耐酸产甲烷菌的分离及其复合菌剂的研究

本文从成都龙泉垃圾填埋场和宜宾造纸厂分离到耐酸性能优良的高温产甲烷菌RY3和中温产甲烷菌SH4，并将其与实验室现有的利用不同底物的产甲烷菌配伍组合成了复合菌剂。采用活性污泥作为固体附着物，研制出了固体产甲烷菌复合菌剂。 菌株RY3的pH耐受范围为5.5～10.5，最适生长pH 6.0～8.0。菌株RY3为革兰氏阳性，长杆状，多数单生，不运动；菌落浅黄色，形状近圆形；利用H2+CO2或甲酸盐作为唯一碳源生长，不利用乙酸盐，对氯霉素非常敏感。该菌最适生长温度为55℃～65℃，最适NaCl浓度为0～2%。根据形态和生理生化特性及16S rDNA序列分析将其初步定为热自养甲烷热杆菌（Methanothermobacter thermautotrophicus）。添加RY3菌液与仅添加厌氧污泥作为接种物相比一周内可使达到最大产甲烷速率所需时间缩短三分之二，甲烷总产量提高约1.8倍。菌株SH4的生长pH范围5.5～9.5，其对酸碱具有良好的适应性，培养3天后，在初始pH值为6.0～8.0的培养基中甲烷产量相差不大，且基本达到最大产量。SH4革兰氏染色阳性，短杆状，多数单生，不运动；菌落近圆形，微黄；利用H2+CO2或甲酸盐作为唯一碳源生长，不利用乙酸盐，对氯霉素非常敏感。SH4最适生长pH 为7.0，最适生长温度为35℃，最适NaCl浓度为0～1.5%。实验表明，添加SH4菌液与仅添加厌氧污泥作为接种物相比可使产甲烷启动时间缩短三分之一，甲烷总产量亦有大幅提高。从形态和生理生化特征以及16S rDNA序列分析表明SH4为嗜树木甲烷短杆菌（Methanobrevibacter arboriphilus）。 以活性污泥为附着物，与培养基和菌种经搅拌后厌氧发酵可得产甲烷菌固体复合菌剂。固体复合菌剂的pH耐受范围为5.5～9.5，温度耐受范围为15℃～65℃，表明其对环境的适应性较强。以猪粪为底物进行厌氧发酵，接种复合菌剂进行试验，以接种实验室长期富集的产甲烷厌氧污泥作为对照，在20℃时，发酵甲烷浓度与对照基本一致，但每日产气量优于对照，第15天时接种复合菌剂的发酵瓶每日产气量是对照的1.59倍；50℃时达到最大甲烷含量所需时间比对照缩短三分之二，三周内总产气量约为对照的2.7倍，甲烷总产量约为2.8倍。以不加接种物为对照，接种复合菌剂20℃时发酵甲烷含量达到50%约需2周，对照2周内甲烷含量最高仅为4.3%；50℃时接种复合菌剂发酵仅需约1周甲烷含量便可达50%，对照则至少需要2周。 In this paper, high-temperature Methanogen RY3 and middle-temperature SH4 were isolated from Chengdu Longquan refuse landfill and Yibin paper mill. They could be used to make compound inoculum that producing methane with the existing Methanogens utilized different substrate. With using anaerobic activated sludge be solid fixture, the process had been designed to produce solid compound inoculum. Strain RY3 possessed excellent capacity of acid and alkali-tolerant. The pH-tolerant scale of RY3 was 5.5～10.5 and its optimum pH value for growth was 6.0～8.0. RY3 was G+, long-rod shape, monothetic and nonmotile, the colony was pale yellow with suborbicular-shape. Formate or H2+CO2 but not acetate was utilized by RY3 as sole C-source, and it was very sensitive to chloramphenicol. Besides, strain RY3 grew fastest at 55℃～65 and 0℃～2% NaCl. Characteristics of modality and physiology with sequence analysis of the 16s rDNA gene of strain RY3 preliminarily showed that it was Methanothermobacter thermautotrophicus. The experiments indicated that the time which began to produce methane with the highest velocity could be shortened two third by adding RY3 in one week, and the total methane production also was 1.8 times than before. Strain SH4 possessed wide scale of growing pH（5.5～9.5）and excellent ability of acclimatizing itself to acid-alkali. The methane production had no apparent difference among those cultivated in different initial pH（6.0～8.0）after three days and equaled to the maximum production basically. Cells of SH4 were G+, short-rod sharp, monothetic and nonmotile. The colony was pale yellow with suborbicular-shape. Formate or H2+CO2 but not acetate was utilized by SH4 as sole C-source, and it was very sensitive to chloramphenicol. Besides, it grew fastest at pH 7.0，55 ℃～65 and 0℃～2% NaCl concentration. The experiment indicated the time that began to produce methane could be shortening one third by adding SH4. And the total methane production also rose apparently. Characteristic of modality and physiology with sequence analysis of the 16S rDNA gene of strain SH4 demonstrated it was Methanobrevibacter arboriphilus. The activated sludge was utilized as fixture, mixed with culture medium and inocolum, that the solid compound inoculum could be produced by anaerobic fermentation. The compound inoculum could grow between pH 5.5～9.5, 15℃～65. It demonstrated the compound inoculum ha℃ve great ability of adapting to circumstance. In the experiment that making pig manure be substrate and taking the anaerobic sludge producing methane that cultured in long term in laboratory to be comparison, the concentration of methane in fermentation added compound inoculum almost equal to the comparison at 20℃, but the volume of gas production could be a little higher. The gas production everyday inoculated compound inoculum was 1.59 times to comparison. The time that the concentration of methane to maximum could be shortening by two third by adding compound inoculum, and the total gas production was 2.7 times to comprison while the total methane production was 2.8 times. If take the no inoculum be the comprasion, anaerobic fermentation added compound inoculum made the concentration of methane to 50% in 2 weeks but the comparison only to 4.3% at 20℃. The time that the concentration of methane to 50% by adding compound inoculum only need 1 week, but the comparison need 2 weeks at 50℃.

Kinetic modeling of growth and biodegradation of phenol and m-cresol using Alcaligenes faecalis

A phenol-degrading. microorganism, Alcaligenes faecalis, was used to study the substrate interactions during cell growth on phenol and m-cresol dual substrates. Both phenol and m-cresol could be utilized by the bacteria as,the sole carbon and energy sources. When cells grew on the mixture of phenol and m-cresol, strong substrate interactions were observed. m-Cresol inhibited the degradation of phenol, on the other hand, phenol also inhibited the utilization of m-cresol, the overall cell growth rate was the co-action of phenol and m-cresol. In addition, the cell growth and substrate degradation kinetics of phenol, m-cresol as single and mixed substrates for A. faecalis in batch cultures were also investigated over a wide range of initial phenol concentrations (10-1400 mg L-1) and initial m-cresol concentrations (5-200 mg L-1). The single-substrate kinetics was described well using the Haldane-type kinetic models, with model constants of it mu(m1) = 0.15 h(-1), K-S1 = 2.22 mg L-1 and K-i1 = 245.37 mg L-1 for cell growth on phenol and mu(m2) = 0.0782 h(-1), K-S2 = 1.30 mg L-1 and K-i2 = 71.77 mgL(-1), K-i2' = 5480 (mg L-1)(2) for cell growth on m-cresol. Proposed cell growth kinetic model was used to characterize the substrates interactions in the dual substrates system, the obtained parameters representing interactions between phenol and m-cresol were, K = 1.8 x 10(-6), M = 5.5 x 10(-5), Q = 6.7 x 10(-4). The results received in the experiments demonstrated that these models adequately described the dynamic behaviors of phenol and m-cresol as single and mixed substrates by the strain of A. faecalis.

Phosphine-Free Synthesis of High-Quality CdSe Nanocrystals in Noncoordination Solvents: "Activating Agent" and "Nucleating Agent" Controlled Nucleation and Growth

CdSe nanocrystals (NCs) are prepared in noncoordination solvents (1-octadecene (ODE) and paraffin liquid) with Ion g-chain primary alkylamine as the sole ligand, ODE-Se, and cadmium fatty acid salt as precursors. The obtained NCs meet the four fundamental parameters for high-quality NCs: high crystallinity, narrow size distribution, moderate photoluminescence quantum yield, and broad range size tunableness. Further, by simply regulating the relative molar ratio of alkylamine to cadmium precursor, the regular sized "nuclei" and final obtained NCs can be produced predictably within a certain size range.

Efficient inverted top-emitting organic light-emitting diodes using ultrathin MoO3/C-60 bilayer structure to enhance hole injection

Efficient inverted top-emitting organic light-emitting diodes with aluminum (Al) as both the cathode and semitransparent anode are investigated. It is found that introduction of the ultrathin molybdenum trioxide (MoO3)/fullerene (C-60) bilayer structure between the low work function Al top anode and the hole-transporting layer dramatically enhances the device performance as compared to the devices with sole MoO3 or C-60 buffer layer. The ultraviolet photoemission spectroscopy and x-ray photoelectron spectroscopy indicate that the hole injection barrier between Al anode and hole-transporting layer is effectively reduced via strong dipole effect at Al/MoO3/C-60 interfaces with its direction pointing from Al to C-60.