提高猪粪和城市垃圾堆肥质量的菌种选择

室内发酵菌剂筛选试验的结果表明：细黄链霉菌(Streptomyces microflavus)、彩色云芝(Palystictus versicolor )和假单胞杆菌属(Pseudomonas sp.)3种菌剂对促进猪粪、城市垃圾腐熟最有利。在有机肥发酵过程中，除臭作用员明显的分别是绿色木霉(Trichoderma viride)、青雷属真菌(Penicillium sp.-1)和细黄链霉菌(Streptomyces mi-croflavus)菌剂。     

生防放线菌剂对魔芋根域微生物区系的影响

为从微生态角度探索接种生防放线菌剂对魔芋根域微生物区系的影响,以生防放线菌娄彻氏链霉菌(Streptomyces rochei)、濑里予链霉菌(S.senoensis)和M(肉质链霉菌S.carnosus和密旋链霉菌S.pctum按质量比1:1的固态发酵混合制剂)为接种剂,采用基质拌菌法接种生防菌剂进行盆栽试验,通过稀释平板法测定魔芋根区、根表土壤和根内放线菌、细菌及真菌数量并采用分子生物学技术对优势真菌和细菌进行分类鉴定.结果显示:(1)3种菌剂接种164 d,魔芋根域检出的接入放线菌活菌数量高达106 CFU g-1以上,根表土壤中放线菌数量较对照增加24.6%-263.1%;(2)供试3种菌剂接种164 d后,魔芋根域真菌数量减少18.9%-100.0%,且接种处理魔芋根域有害优势真菌腐皮镰孢菌(Fusarium solani)和红球丛赤壳菌(Nectria haematococca)较未接种对照大幅度降低;(3)不同菌剂接种处理,魔芋根区土壤和根内细菌数量较对照减少21.5%-73.3%,但根内芽孢杆菌(Bacillus sp.)数量增加414.0%-1015.0%.本研究表明供试生防菌具有较稳定的定殖能力,亦能改善魔芋根域土壤微生物区系.     

基于木质素生物降解的堆肥化接种剂开发

选取由农林废物堆肥中筛选出的木质素降解优势土著微生物枯草芽孢杆菌(Bacillus subtilis)、铜绿假单孢菌(Pseudomonas aeruginosa)、黑曲霉(Aspergillus niger)、简青霉(Penicillium simplicissim)、栗褐链霉菌(Streptomyces badius),依据PLFA-PLS定量分析所得堆肥化2次发酵期有效的木质素降解微生物群落组成比例混合接种至稻草基质发酵瓶中,做1组L9(34)正交试验以优化混合比例,期望开发1种基于木质素降解的高效堆肥化接种剂.试验结果表明:混合菌剂具有较强的木质素降解能力,其对木质素的降解是木质素过氧化物酶、锰过氧化物酶、漆酶、纤维素酶和半纤维素酶共同作用的结果;当按照个数比细菌∶放线菌∶真菌为85∶5∶ 10,枯草芽孢杆菌∶铜绿假单孢菌为55∶25,黑曲霉∶简青霉为2∶1配比时,木质素、纤维素、半纤维素降解率最高,分别达到22.13%,48.97%和55.93%;在不灭菌前提下,按此配比接入菌剂,其木质素、纤维素、半纤维素降解率分别比不接菌剂发酵稻草提高19.16,38.25和46.30百分点.     

一株抗青枯假单胞菌放线菌的筛选、鉴定及发酵条件优化

从河南中牟土壤中筛选出一株对青枯假单胞菌(Pseudomonas solanacearum)有较强拮抗作用的放线菌ZM-16,它对几种常见病原细菌和真菌都有较强的抑制效果,其发酵液抗菌谱较广.通过菌株形态特征、培养特性、生理生化特性、化学特征研究以及16SrRNA序列分析,初步确定菌株ZM-16为链霉菌属的抗生链霉菌(Streptomyces antibioticus).通过发酵优化实验,确定该菌株的最佳发酵配方为:蔗糖8%、大豆粉5%、NaNO31%、NaCl0.2%、CaCO30.4%、K2HPO40.08%;最佳发酵条件为:发酵培养基初始pH值7.0,30℃,恒温摇床200r/min,培养6d,抑菌效果最好.     

拮抗放线菌T111菌株鉴定、发酵液理化性质测定及发酵条件优化

为明确放线菌T111在植物病害生物防治中的应用潜力,通过形态观察、生理生化测定结合16S rDNA序列分析确定了其分类地位,采用平板打孔法测定了发酵液的理化性质,并采用单因素试验与正交设计方法筛选出该菌株的优化发酵配方和发酵条件.结果表明,T111与黄色链霉菌Streptomyces flaveus strain NBRC3359(AB184749)序列的同源性最高,初步鉴定为黄色链霉菌(Streptomyces flaveus).该发酵液对黄瓜菌核、黄瓜蔓枯、棉花红腐病菌等有较高的抑制活性;贮存稳定性和热稳定性良好,4℃贮存60 d,抑菌活性为对照的88.3%,80℃处理60 min抑菌活性为对照的90.9%;对pH值较敏感,在pH 5～7时活性较高;对紫外线稳定,紫外光照射12 h,其活性基本不受影响.培养基优化组合为:可溶性淀粉20 g,大豆蛋白胨5 g,NaNO3 20 g,FeSO4 0.5 g,去离子水1 000 mL;优化发酵条件为:发酵温度27℃,发酵时间5 d,初始pH 6.0,接种量5%,装液量75 mL/250 mL,摇床转速150 r min-1.图6表2参18     

秸秆干发酵沼气增产研究

研究了复合菌剂预处理秸秆和添加沼气发酵促进剂对秸秆干发酵的影响.结果表明,秸秆经菌剂预处理后的产气量比不加菌剂预处理(对照)提高29.54%;在菌剂预处理过的秸秆中加入促进剂,其产气量比对照提高35.28%,比单纯菌剂预处理提高4.43%.另外,菌剂预处理过的秸秆的TOC降解率和纤维素降解率分别比对照高136.32%和47.68%;秸秆经菌剂预处理后再添加促进剂的TOC降解率、纤维素降解率分别比对照高169.58%和49.62%,比单纯菌剂预处理高14.07%和3.36%.图2表1参8     

化感链霉菌6803菌株液体发酵工艺优化的研究

土壤微生物链霉菌6803菌株被证明对高等植物具有化感作用。采用单因子实验方法,研究液体发酵碳源、氮源、无机盐、发酵温度、发酵液初始pH值、摇床转速、发酵时间等对链霉菌6803菌株菌丝体产量及发酵液化感作用的影响,用均匀试验设计法优化了发酵工艺条件。结果表明：链霉菌6803菌株生长和产生化感作用的最佳发酵条件为：p（淀粉）=26．67g·L-1,p（蔗糖）=25．15g·L-1,p（NH4C1）=0．30g．L-1,黄豆饼粉浸液8．88％,所（NI-h）H2P04]=O．18g·L-1,p（FeS04．7H20）=O．002g·L-1,p（NaCl）=0．75g·L-1,pH值7．6,装量系数O．16,接种量3％,温度36℃,转速200r·min-1,发酵144h。本研究为利用微生物次生代谢产物作为天然除草剂奠定了基础。     

3种菌剂对水稻秸秆降解性能的影响

为寻求秸秆的高效快速处理途径,采用EM、VT1000、强兴3种菌剂作用于秸秆与果皮蔬菜混合垃圾按不同配比混合的物料,无需发酵,通过分析不同菌剂处理后秸秆混合物料的理化性质、降解性能和微生物种群结构变化,探讨不同菌剂对秸秆降解性能的影响。结果表明,3种菌剂均对物料的全碳、全氮、C/N具有降低作用,可提高速效磷和速效钾的含量,加快半纤维素的降解。EM对全碳、速效磷和C/N的作用效果较好,VT1000对全氮、速效钾和半纤维素的作用效果较好,且各菌剂于物料配比3?2时处理效果最佳,EM菌剂处理后物料全碳质量分数和C/N比值分别为8.19%和14.44,速效磷上升幅度为51.49%;VT1000菌剂处理后物料全氮下降幅度为55.24%,速效钾上升幅度为83.60%,半纤维素降解率为60.09%。采用高通量测序技术分析微生物群落结构对物料性能的影响,结果表明,不同菌剂处理后物料的优势细菌菌群为Proteobacteria(变形菌门)、Actinobacteria(放线菌门)、Bacteroidetes(拟杆菌门)、Acidobacteria(酸杆菌门)和Chlorobacteria(绿弯菌门),优势真菌菌群为Ascomycota(子囊菌门)、Zygomycota(接合菌门)、Basidiomycota(担子菌门)和Unclassified-k-Fungi(未经分类的真菌)。研究发现,优势菌群的相对丰度是影响秸秆性质变化的主要原因,EM菌剂处理的物料,优势菌群Unclassified-k-Fungi的相对丰度明显高于其他菌剂的处理,这直接影响物料的C/N变化;而VT1000菌剂处理的物料,优势菌群Acidobacteria相对丰度明显高于其他菌剂,其与物料中全氮和速效钾存在一定的相关性。该研究可为探索菌剂复配或研发新型菌剂用于秸秆的微生物降解奠定理论基础。     

反硝化聚磷菌B8干粉菌剂的制备及应用

为获得反硝化聚磷菌(DNPAOs)干粉菌剂,以反硝化聚磷菌B8液态菌剂为菌种来源,以麦麸、玉米干粉混合物作为载体,以牛肉膏蛋白胨培养基为发酵液,进行增菌培养.在麦麸、玉米干粉载体配比(W/W)为85%∶15%,投菌液量为4 m L·g~(-1)干粉,发酵液用量为4 m L·g~(-1)干粉的条件下制得干粉菌剂.其使用效果为:(1)当主要污水处理因子为硝酸盐氮时,干粉菌剂成品的最佳发酵液p H值为5.5,该菌剂对化学需氧量、硝酸盐氮和总磷的去除率可达43.26%、92.76%和65.77%;(2)当主要处理因子为总磷时,则同等条件下所制得的干粉菌剂成品的最佳发酵液p H值为6.5,该菌剂对化学需氧量、硝酸盐氮和总磷的去除率可达44.41%、56.21%和87.30%.扫描电镜显示B8菌株附着于菌剂载体上,吸收培养液的营养用于繁殖生长及发酵,干燥后B8可固定于载体上,再将其投放于待处理废水中即可发挥其菌效.     

Streptomyces sp. WJS-825产谷氨酰胺转胺酶发酵条件优化及中试

应用五因子二次正交旋转回归试验设计,建立了微生物谷氨酰胺转胺酶 (TG)发酵生产过程中以酶活力和菌体细胞生长量作为目标函数的数学模型,并以此模型对链霉菌 (Streptomycessp. )WJS-825菌株发酵生产TG的培养条件进行优化,确定了影响TG生产的主要因子及其最适取值为多价胨 2. 1%、可溶性淀粉 1. 5%、初始pH 7. 0及培养温度 30℃.以该优化工艺条件进行了 5L发酵罐小试和 200L发酵规模的中试生产.结果表明,在中试发酵生产中使用以豆饼粉部分替代多价胨的经济性发酵优化培养基,以及发酵过程中在线监控pH、溶氧系数等多项发酵调控参数,并分段控制pH、温度、通气量和搅拌转速以及进行适时的流加补充碳源,该菌株生长繁殖能力强、产酶效果好,TG活性达 3. 2u/mL,而且连续重复的中试发酵生产试验的TG产量均稳定在 3. 2u/mL以上. 图 2表 3参 15     

链霉菌的抗砷特性及其对蜈蚣草富集砷的作用

本文研究了链霉菌Streptomyces sp.的耐砷特性及其对蜈蚣草富集砷的影响。结果表明,Streptomyces sp.可在100mmo·lL-1的砷酸盐溶液中生长,具有较强的抗砷毒害能力,且在48h内对As(Ⅴ)的还原率达96.5%。施用Streptomyces sp.能促进植物对砷的吸收,蜈蚣草地上部砷浓度为930mg·kg-1,地上部砷累积量达到对照组的2.09倍。加入Streptomyces sp.后,能促进根际土壤中As(Ⅴ)还原成As(Ⅲ),大幅度降低根际土壤残渣态砷含量,从48.15mg·kg-1下降至28.75mg·kg-1。Streptomyces sp.通过影响蜈蚣草根际环境,提高根际土壤pH,增加DOC含量,促使砷形态变化,从而增加砷生物可利用性。该菌可作为强化蜈蚣草修复砷污染土壤的材料。     

Screening of the marine actinomycetes with anti-complement activity and isolation of the active fractions北大核心CSCD

Studies on anti-complement agents are essential for the development of drugs to treat various diseases caused by excessive or abnormal activation of the complement system. However, studies on microbial-derived anti-complement agents are still very limited. The anti-complement activities of 42 marine-derived actinomycete strains isolated from the sediment samples collected from Xinghai Bay in Dalian were studied using the hemolysis method. In addition, the active fractions of the strain Streptomyces sp. DUT11 were separated by C18 and Sephadex LH-20 column chromatography, and the active compounds with anti-complement activity were identified. Streptomyces strains S187, M5, S088, M8, S063, DUT11 and MD16 showed good anti-complement activities upon being cultured in TSB medium, with the strain DUT11 showing the best anti-complement activity among these strains. When different fermentation media were tested, the highest anti-complement activity of DUT11 was observed in M33 fermentation medium. The anti-complement activity was 56.5% for the extract of supernatant and 60.8% for the extract of mycelia, respectively. Furthermore, three compounds with anti-complement activity were obtained from the active components, which were identified as 3-indolecarboxylic acid (1), p-hydroxy benzoic acid (2) and 2-pyrrolecarboxylic acid (3). These results demonstrated that marine-derived streptomycetes can be employed to produce active compounds with anti-complement activity. This study presents a new alternative for the utilization of marine actinomycetes and provides a basis for the exploration of new anti-complement agents. © 2018 Science Press. All rights reserved.     

高产黑色素菌株的分离及鉴定

对从土壤中分离出的321株菌株进行了筛选,得到1株高产胞外黑色素的菌株,比较了其在不同培养基上产黑色素的能力.初步确定该菌株为链霉菌属.该菌黑色素产量高,约为0.70g/L,在产黑色素的微生物中,链霉菌可以作为一类新的菌种资源.图2表1参15     

中国红豆杉内生放线菌En-1的分离、鉴定及其次生代谢物的研究

采用选择性分离方法对药用植物红豆杉Taxus chinensis嫩枝和叶进行内生菌分离,得到一株放线菌En-1,经鉴定为链霉菌Streptomyces sp.;并对该菌进行液体培养条件优化、次生代谢物初筛及其抗菌活性的研究。通过添加宿主植物浸出物至En-1液体培养中以考察宿主对后者生长的影响,结果表明红豆杉叶浸出物能促进En-1的体外生长;而且En-1次生代谢物对5株供试菌中的黑曲霉具有抑制活性。通过En-1菌培养基优化并联合波谱法检测其发酵产物,显示8#配方（淀粉25 g.L-1,KNO3 1 g.L-1,K2HPO4.3H2O 0.5 g.L-1,NaCl 0.5 g.L-1,FeSO4.7H2O 0.01 g.L-1,MgSO4.7H2O 0.5 g.L-1）培养液中,En-1所产的次生代谢物最具结构多样性;同时8#配方也有利于该菌产生抗菌活性产物。本研究提示红豆杉内生放线菌可作为寻求药物先导化合物的资源菌。     

Isolation and characterisation of antibiotic producing microorganisms in composted Nigerian soil

Compost samples obtained from different locations within the premises of the university of Lagos were analysed to determine the presence and types of antibiotic-producing bacteria, fungi and actinomycetes using nutrient agar, potato Dextrose agar and starch casein nitrate agar respectively as culture media. A variety of bacteria were isolated and these included Staphylococcus aureus, B. subtilis, B. pumilis, B. lactesporus, B. megaterium, B. pulvifaciens, B. licheniformis, Streptococus spp., Corynebacterium spp. and E. coli. The fungal isolates encountered were Aspergillus niger, A. flarus, T. viridae, P. chrysogenum, P. pinofylum and Absida spp., while the following actinomycetes were identified: Norcadia spp., Micromonospora spp., Streptomyces scabies, S. reticuli and S. hygroscopicus. When these organisms were screened for antibiosis, the following species were found to be antibiotic producers: B. licheniformis, B. subtilis, Penicillium chrysogenum, Streptomyces reticuli, S. hygroscopicus and Micromonospora spp. The fungus Penicillium chrysogenum had the highest rate of antibiotic production with an inhibitory zone width of 17mm while Trichoderma viridae produced toxins lytic to other fungal hyphae.     

Biochemical composition of cephalopods with different life strategies,with special reference to a giant squid,<Emphasis Type="Italic"> Architeuthis</Emphasis> sp.

The present study is a comprehensive comparison of the biochemical composition (protein, lipid, glycogen and cholesterol contents, and amino acid and fatty acid profiles) of 14 species of cephalopods with different life strategies (benthic, nektobenthic, benthopelagic and pelagic) in distinct habitats (neritic, demersal, oceanic and deep sea), with special emphasis placed on a male Architeuthis sp. The giant squid showed a significantly lower protein and total amino acid content in the gonad and digestive gland. The major essential amino acids were leucine, lysine and arginine. The major non-essential amino acids were glutamic acid, aspartic acid and proline. The benthopelagic cirrate octopus Opisthoteuthis sp. showed a lower content of these nitrogen compounds in the muscle. In respect to lipid and fatty acid contents, the giant squid showed the highest values in the gonad and digestive gland and the lowest in the muscle. Most of saturated fatty acid content was presented as 16:0; monounsaturated fatty acid content, as 18:1 and 20:1; and polyunsaturated fatty acid content, as 20:4n-6, 20:5n-3 and 22:6n-3. The highest cholesterol content in the gonad was detected in Opisthoteuthis sp. and Architeuthis sp.; in the digestive gland. in Todarodes sagittatus; and in the muscle, in Sepia elegans. The highest glycogen value in the gonad was detected in Octopus vulgaris; in the digestive gland and muscle, the highest values were attained in Opisthoteuthis sp. The relationships between life strategies and biochemical composition were investigated and principal component analysis (PCA) was performed. The different cephalopod life strategies could be well separated on the basis of the first two principal components. The nektobenthic and pelagic strategies were clearly separated from the benthic, suggesting that these groups of species are characterized by lower lipid content and higher protein content in the gonad. A rather similar life-strategy distinction was obtained for the digestive gland. The benthopelagic strategy was also well separated from benthic and pelagic strategies and from Architeuthis sp. In the muscle, the results indicated lower nitrogen and carbon compound contents in Architeuthis sp. and Opisthoteuthis sp. The environmental conditions that possibly explain the Architeuthis sp. stranding, the relationships between biochemical compositions and the life strategies of the different cephalopod species studied are discussed.Communicated by S.A. Poulet, Roscoff     

中国5种土壤跳虫对重金属镍的毒性响应

为了研究镍(Ni~(2+))对不同种类跳虫的毒性效应,将曲毛裸长(Sinella curviseta)、四刺泡角(Ceratophysella duplicispinosa)、小原等节(Proisotoma minuta)、茉莉花长角(Entomobrya sp.)、符氏直棘(Orthonychiurus folsomi)5种中国优势跳虫物种,在琼脂培养基环境下分别暴露于Ni~(2+)的8个浓度组中进行24 h、72 h-LC_(50)急性毒性试验和28 d生存试验。利用Bliss法测得其5种跳虫的24 h-LC_(50)分别为52.99、35.91、33.46、23.72、13.50 g·L~(-1);72 h-LC_(50)为46.25、4.54、13.37、14.79、4.35 g·L~(-1)。在28d生存试验中得到成虫存活数量和繁殖情况。结果表明,5种跳虫中曲毛裸长对镍有较强的耐受性,茉莉花长角和小原等节次之,四刺泡角和符氏直棘较弱。符氏直棘与四刺泡角相比,随时间增加对镍的耐受性有相对增强的趋势。此外,Ni~(2+)对跳虫的毒性影响表现出阶段性的阈值效应。Ni~(2+)对5种跳虫的繁殖均有抑制作用。     

Comparison of growth and lipid accumulation properties of two oleaginous microalgae under different nutrient conditions

This study compared the growth and lipid accumulation properties of two oleaginous microalgae, namely, Scenedesmus sp. LX1 and Chlorella sp. HQ, under different nutrient conditions. Both algal species obtained the highest biomass, lipid content and lipid yield under low-nutrient conditions （mBGll medium）. The biomass, lipid content and lipid yield of Scenedesmus sp. LX1 were 0.42g·L^-1, 22.5% and 93.8mg·L^-1, respectively. These values were relatively higher than those of Chlorella sp. HQ （0.30g·L^-1, 17.1% and 51.3mg·L^-1, respectively）. These algae were then cultivated in an SE medium that contained more nutrients; as a result, the biomass and lipid yield of Scenedesmus sp. LX1 reduced more significantly than those of Chlorella sp. HQ. Opposite results were observed in lipid and triacylglycerols （TAGs） contents. The cell sizes of both algal species under low-nutrient conditions were larger than those under high-nutrient conditions. Chlorella sp. HQ cells did not aggregate, but Scenedesmus sp. LX1 cells flocculated easily, particularly under low-nutrient conditions. In summary, low-nutrient conditions favour the growth and lipid production of both algae, but Scenedesmus sp. LX1 outperforms Chlorella sp. HQ.     

Synergistic degradation of pyrene and volatilization of arsenic by cocultures of bacteria and a fungus

The combination of two bacteria (Bacillus sp. PY1 and Sphingomonas sp. PY2) and a fungus (Fusarium sp. PY3), isolated from contaminated soils near a coking plant, were investigated with respect to their capability to degrade pyrene and volatilize arsenic. The results showed that all strains could use pyrene and arsenic as carbon and energy sources in a basal salts medium (BSM), with the combined potential to degrade pyrene and volatilize arsenic. Bacillus sp. PY1, Sphingomonas sp. PY2 and Fusarium sp. PY3 were isolated from the consortium and were shown to degrade pyrene and volatilize arsenic independently and in combination. Fungal-bacterial coculture has shown that the most effective removal of pyrene was 96.0% and volatilized arsenic was 84.1% after incubation in liquid medium after 9 days culture, while bioremediation ability was 87.2% in contaminated soil with 100 mg·kg^-1 pyrene. The highest level of arsenic volatilization amounted to 13.9% of the initial As concentration in contaminated soil after 63 days. Therefore, a synergistic degradation system is the most effective approach to degrade pyrene and remove arsenic in contaminated soil. These findings highlight the role of these strains in the bioremediation of environments contaminated with pyrene and arsenic.