溶藻细菌的分离及溶藻特性研究

从爆发蓝藻水华的鱼池采集水样,以铜绿微囊藻为对象进行溶藻细菌筛选,得到R1、R2、R3、R5、R6、R8和R9共7株溶藻细菌,生理生化特征初步鉴定R5为海鱼弧菌,另6株为非发酵型细菌.R1、R3、R5、R8和R9细菌含量为105～108 cfu/ml时,对微囊藻的抑制率很高,效果优于R2和R6.对R1、R3、R5和R9...     

Identification of marine algicidal Flavobacterium sp. 5 N-3 using multiple probes and whole-cell hybridization

ABSTRACT: We designed five 16S rRNA-targeted oligonucleotide probes (Sp probes) specific for Flavobacterium sp. 5 N-3, which inhibits the growth of red tide phytoplankton Gymnodinium mikimotoi (Dinophyceae). These probes were evaluated by whole-cell hybridization against 5 N-3 cells incubated under laboratory conditions. The fluorescence signal from the cell detected with Sp probe mix5, a mixture of the five probes, was 8.4-fold higher than that obtained with only one Sp probe (Sp01RF). The signal obtained by this method was strong enough to recognize 5 N-3 cells and count them under the epifluorescence microscope, while the signal was often undetectable when a single probe was used. Fluorescence intensities of cells at stationary phases and of 'starved' cells in sterile seawater using Sp probe mix5 were low but still sufficient for enumeration. These Sp probes did not hybridize with 11 strains from the Cytophaga/Flavobacteria/Bacteroides phylum and did react with strain 5 N-3 following whole-cell hybridization. These results show that 5 N-3 cells cultivated under our laboratory conditions can be detected by whole-cell hybridization with the five designed probes. These data also suggest that this technique may be useful for detection of an algicidal bacterium 5 N-3 in the natural environment.     

秸秆饲料化全细胞纤维素酶的构建及应用

为提升秸秆饲料的品质,通过构建全细胞酿酒酵母纤维素酶,探究全细胞纤维素酶用于秸秆饲料化的应用前景。通过与其他青贮菌剂对比用于秸秆饲料化发酵,比较分析全细胞酿酒酵母纤维素酶在秸秆饲料化中的应用效果。试验发酵条件设置为：发酵温度28℃,发酵时间14 d,真空厌氧发酵。结果表明：与乳酸菌发酵相比,纤维素酶与乳酸菌混合发酵的玉米秸秆铵态氮/总氮比没有明显上升（P>0.05）,因此不会造成饲料中的蛋白过量分解;纤维素含量显著下降（P<0.01）,相比未发酵秸秆下降15.03个百分点,比乳酸菌发酵效果提高一倍以上,与商用青贮剂发酵效果相当;半纤维素含量相比未发酵秸秆下降2.70个百分点。研究表明酿酒酵母全细胞纤维素酶与乳酸菌混合处理秸秆,可提高纤维素酶酶解效率,有效减低秸秆中纤维素、半纤维素含量,酶解产物可加速乳酸菌发酵进程,全细胞酶可增加秸秆单细胞蛋白含量,实现玉米秸秆发酵效果、营养价值和利用效率的同步提升。     

高效溶藻菌对富营养化水体的治理效果

为研究自然条件下溶藻菌RZ14菌株治理水华的效果,在调查泉城公园景观水体富营养化状况的基础上,选取水华较严重的映日湖作为该溶藻菌剂原位生物控制的测试水域,对该湖区叶绿素a、有机质、氨氮及总磷变化进行跟踪测定。结果表明,映日湖水华现象严重,未达到一般景观用水V类标准,属重度营养化。经该溶藻菌剂生物治理后,水华消除,叶绿素a含量低于0.05 mg/L,CODCr清除率50.01%,BOD5清除率73.38%,氨氮清除率70.51%,总磷清除能力88.59%。通过原位试验,表明RZ14抑藻剂能快速消除水华现象,对富营养化水体治理效果显著,可开发为溶藻菌剂应用于蓝藻水华的治理。     

溶藻细菌及其作用物质研究进展

随着全球气候变化和水体富营养化程度加剧，蓝藻水华已经成为一个世界性的环境问题。它不但对水生生态系统有许多负面影响，还会严重威胁人类的公共卫生安全。目前，蓝藻水华的治理技术主要包括物理法、化学法和生物法等。近年来，溶藻细菌作为一种新型蓝藻水华生物防治方法，由于其环境友好、作用特异受到广泛关注。目前，已经发现了很多溶藻菌，并开展了溶藻物质鉴定和溶藻作用机制研究。本文根据近十年来发表的文献，对溶藻细菌种类、溶藻物质作用机理的研究现状进行了归纳、总结，并对未来进一步研究和应用溶藻菌控制蓝藻水华的进行了展望。     

来源于腌干鱼的乳酸菌中抗氧化酶及胞外多糖研究

为研究从腌干鱼中获取的乳酸菌在细胞内外起抗氧化作用的主要活性物质,通过测定菌株胞内的超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)活性以及提取胞外多糖(EPS)并测定其抗氧化性,利用超高效液相色谱法(UPLC)分析其结构。结果显示,9株菌的SOD、GSH-Px和CAT的最高活性分别为44.67、15.26和1.23 U/mg prot,对应菌株为L21、L4和L21;9株菌的EPS产量为(5.71±0.18)~(50.33±1.89)mg/L,L21的EPS最高纯度为56.16%±1.08%。L21中EPS的DPPH自由基清除能力和还原能力较高,综合抗氧化能力相对较强,结构分析发现其含有甘露糖(Man)、鼠李糖(Rha)、葡萄糖(Glu)和半乳糖(Gal)等4种单糖。研究表明,分离自腌干鱼的乳酸菌中,抗氧化能力越强的菌株细胞内表现出越高的抗氧化酶活性,同时细胞外EPS的产量和纯度也较高,并且EPS也具有较强的抗氧化能力。从中筛选出一株综合抗氧化能力最强的L21,可以进一步作为生物源性抗氧化剂。     

一株铜绿微囊藻溶藻菌的分离鉴定和溶藻特性

以铜绿微囊藻为溶解对象,从富营养化水体中分离到1株溶藻菌H1,通过形态特征及16S rDNA序列分析,该菌株属寡养单胞菌属(Stenotrophomonas sp.),GenBank登录号为KU359254。探讨了菌株H1对铜绿微囊藻(FACHB-1326)的溶藻方式,菌藻浓度、菌的生长时期及光照、pH、温度等环境因子对铜绿微囊藻溶藻效果的影响。结果表明,菌株H1溶藻方式主要是间接作用溶藻。处于对数期的菌株H1抑藻效果最高,溶藻率达77.9%;菌液浓度达10~8 CFU·mL~(-1)以上,溶藻率最高,为81.8%;在pH7、30℃条件下,活性较强;pH7时30℃条件下,溶藻率为72.5%,63.2%;菌株H1对微囊藻生长的抑制效果是全黑条件光循环条件全光条件,全黑条件下溶藻率最高为59.9%。     

Purification and characterization of the extracellular algae-lysing components from an algae-lysing bacteria

Purification, characterization and initial identification of extracellular algae-lysing constituents of fermentation products of the algicidal bacteria S7 are carried out by ultraviolet spectrum, infrared spectrum, fluorescence spectra and mass spectrum, and the results show that the eluates of methanol have the best algicidal effects. These active algicidal substances made of conjugation unsatuated bonds, hydroxyl, saturated alkyl and aromatic ethers. In the three-dimensional fluorescence spectroscopy, two strong fluorescence peaks appear at 264/436 nm and 378/369 nm respectively. Mass spectrum indicates that the active substances consist of six monomers, and their molecular weights are 163.1, 246.3, 274.3, 318.3, 404.5 and 475.4, respectively.     

一株水华鱼腥藻溶藻细菌的分离鉴定和溶藻特性

从不同来源的水体和土样中,筛选出对水华鱼腥藻具有溶藻作用的溶藻菌株SLW6。对菌株SLW6进行形态特征分析和生理生化鉴定,及16S rDNA序列分析,并研究了溶藻菌在不同培养时期、不同pH条件下对菌株SLW6溶藻效果的影响,探究了该菌株对水华鱼腥藻的溶藻方式。结果表明：菌株SLW6呈革兰氏阴性,V-P试验、明胶液化试验、甲基红试验为阴性,过氧化氢试验、硝酸盐还原试验都为阳性,菌株SLW6与产碱杆菌属的Alcaligenes faecalis(NR113606.1)的同源性达到99.65%,结合形态学特征,生理生化鉴定和16S rDNA序列分析,初步判断溶藻菌株SLW6为产碱菌属（GenBank登录号为OP363821）。超声辅助热乙醇提取叶绿素a,分光光度法测定叶绿素a含量,用叶绿素a含量变化计算溶藻率,结果表明SLW6处于对数期时的溶藻效果最好,溶藻率达89.45%;在pH=7条件下,溶藻效果较好,溶藻率为47.03%;溶藻菌SLW6对水华鱼腥藻的作用方式是直接溶藻为主间接溶藻作用为辅。