四环素降解菌的选育、鉴定及其降解特性

生产四环素过程中产生大量发酵底物(俗称"药渣"),采用生物二次发酵法可消除药渣中四环素,使其能作为饲料原料使用.本研究从长期堆放四环素药渣的土壤中筛选四环素降解菌株,经驯化富集后筛选得到TD2和TD3两株能高效降解四环素的菌株.根据表型特征、生理生化特性及16S rDNA序列同源性分析,鉴定TD2为缺陷短波单胞菌(Brevundimonas diminuta),TD3为人苍白杆菌(Ochrobactrum anthropi).TD2和TD3均可利用四环素作为碳源生长.TD2在碳源、氮源、矿物质分别为无碳源、蛋白胨0.5%、CuSO4 0.015%时,降解性能最优;而TD3在葡萄糖0.5%、牛肉膏1.5%、CuSO4 0.015%的培养基中降解效率最大.两株菌的最适培养条件相同:培养时间5 d、温度30℃、接种量1%,且四环素降解率与通气量成正相关.在最适条件下,TD2和TD3的四环素降解率均达90%以上.本实验筛选所得的两株四环素降解菌株可作为四环素药渣进行二次发酵的候选菌株.     

乙草胺降解菌Y-4的分离鉴定及降解特性研究

从长期经乙草胺污染的污泥中分离到一株能以乙草胺为唯一碳源和能源生长的菌株Y-4,通过生理生化实验和16S rDNA同源性序列分析,鉴定为申氏杆菌属（Shinella sp.）。采用室内培养方法,研究了Y-4对乙草胺的降解特性。结果表明,Y-4能有效地降解浓度为5～200 mg.L-1的乙草胺,在48 h内对50 mg.L-1乙草胺的降解率达到83.3%。菌株Y-4降解乙草胺的最适pH值为8.0,最适温度为30℃,其对丙草胺和丁草胺等农药也有良好的降解效果。     

四环素类抗生素降解菌的分离与鉴定

为了获得四环素类抗生素的高效降解菌,从长期受四环素类抗生素污染的土壤中分离、重复驯化筛选得到6株对四环素类抗生素具有降解能力的菌株;采用高效液相色谱法研究了6株降解菌的降解效果,并通过形态、生理生化特征及16S rDNA序列分析对其进行鉴定。结果表明,菌株TJ-2#对土霉素、四环素和金霉素3种四环素类抗生素的降解效果最好,降解率分别为58.3%、63.9%和65.5%,TJ-6#其次;6株降解菌均能以四环素类抗生素作为唯一碳源生长。经形态、生理生化特征及16S rDNA鉴定,菌株TJ-2#为木糖氧化无色杆菌(Achromobacter sp.),TJ-6#为枯草芽胞杆菌(Bacillus sp.)。本研究结果对四环素类抗生素的生物降解具有一定的现实意义。     

一株三唑磷降解菌mp-4的分离鉴定及降解特性的研究

从长期经有机磷农药污染的土壤中分离到一株能高效降解三唑磷的菌株mp 4,通过生理生化实验和16 S rDNA同源性序列分析将该菌鉴定为苍白杆菌属(Ochrobactrum sp.)。mp 4菌能以三唑磷为唯一碳源生长,对三唑磷的降解率为98. 3%。在25~37℃、pH值为6.6时生长较好,27~32℃、pH7 5~8 8时有较好的降解性能。在水稻大田试验中,米壳的三唑磷残留去除率为91 9%,糙米的三唑磷残留去除率为100%。     

多菌灵降解菌的筛选及降解特性初步研究

实验采用富集培养及划线纯化法,从多年生产多菌灵的原药厂下水道污泥中分离筛选到一株可有效降解多菌灵的细菌,经形态观察、生理生化实验及16S rDNA序列分析鉴定为红平红球菌属(Rhodococcus erythropolis)。该菌在温度30℃,pH6.0,补加葡萄糖、酵母浸粉做碳氮源的条件下,150 r/min摇床培养72 h,对多菌灵的降解率高达90.07%。用菠菜作盆栽试验,结果表明,L-1菌悬液可在6 d内有效降解多菌灵使其残留量达到国家相应标准,因而可以将其应用于多菌灵污染的生物修复。     

土霉素降解菌筛选及降解特性研究

【目的】 近年来,抗生素在畜禽及水产养殖中的使用量增加,导致固体废弃物和污水中存在大量的抗生素和耐药菌。土霉素作为用于养殖业主要的抗生素之一,在畜禽粪便和污水中的残留含量较高,因此,筛选并鉴定了能降解残留土霉素的微生物。 【方法】 采用富集驯化法,以菌肥、药渣和畜禽粪便为原料,采用摇床震荡的方法进行微生物培养,采用高效液相色谱法 (HPLC) 进行土霉素含量测定,筛选出能够高效降解土霉素的微生物。本研究还对降解菌在不同温度、pH、转速和接种量条件下的土霉素降解效果进行优化,并最终利用16S rDNA的方法鉴定菌种。 【结果】 筛选出一株能够高效降解土霉素的菌株T4菌,经16S rDNA测序鉴定该菌株为假单胞菌 (Pseudomonas sp.) ,该菌株在30℃时对土霉素的降解率最高,达到了26.75%;不同pH梯度下,T4菌在pH为7时对土霉素的降解率达到最高,为27.03%;转速为150 rpm和170 rpm时,T4菌对土霉素的降解率分别为26.18%和25.59%,考虑到摇床高转速耗能高的因素,因此选择150 rpm为优化的转速;接种量对T4菌降解土霉素的影响较小,而且二者之间呈负相关,接种量1%时降解率最高,为26.88%。优化条件下,T4菌对100 mg/L土霉素的降解率为26.29%;堆肥试验表明,添加了T4菌之后,土霉素去除率更高,为93.21%。 【结论】 本研究筛选出的菌株T4对土霉素有较好的降解能力。通过16S rDNA基因序列分析,T4菌属于假单胞菌 (Pseudomonas sp.),其降解土霉素的优化条件为温度30℃、pH 7.0、转速150 rpm、接种量1%。在堆肥中接种T4菌后,提高了对土霉素的去除作用,表明T4菌作为土霉素降解菌具有污染治理的潜力。      

功夫菊酯降解菌 GF-1 的分离鉴定及其降解特性研究

从农药厂废水处理池的活性污泥中分离到一株功夫菊酯高效降解菌 GF-1,根据表型特征、生理生化特性和 16S rDNA 序列同源性分析,将 GF-1 鉴定为芽孢杆菌属(Bacillus sp.).GF-1 能以功夫菊酯为唯一 C 源进行生长,72 h 对100 mg/L的功夫菊酯的降解率达 85%.GF-1 降解功夫菊酯的最适温度为 30℃,最适 pH 为 7.0,该菌降解功夫菊酯的速率与初始接种量呈正相关.GF-1 投加土壤,可以提高土壤中功夫菊酯的降解速率.通过吖啶橙与升温法对质粒消除,结果表明 GF-1 的降解功能与质粒相关.     

一株新的多菌灵高效降解茵的筛选与降解特性分析

从长期施用多菌灵农药的土壤中,通过富集筛选,获得1株新的多菌灵高效降解菌株。通过生理生化实验和16SrDNA序列同源性分析鉴定该菌株,应用高效液相色谱法对纯培养条件下菌株的降解特性和粗酶提取液的降解性能进行了分析。结果表明,筛选所获得的菌株与Raoultella菌属的亲缘关系最近,将其命名为Raoultellasp．MBC,该菌株能在以多菌灵为唯-碳源的无机盐培养基中生长;25℃、pH7．0、200r·min。的最适生长条件下避光振荡培养72h,多菌灵的降解率达到100％;在最适培养条件下外加氮源和碳源在培养后期均可以提高多菌灵的降解率,外加氮源对多菌灵的降解效果优于外加碳源;该菌体的粗酶提取液具有降解多菌灵活性,且多菌灵降解酶为诱导酶。研究结果为多菌灵污染土壤的生物修复和酶修复提供了材料和理论依据。     

污染土壤中原位阿特拉津降解菌的分离和鉴定

为克服传统富集培养分离降解菌的局限性,直接将长期受阿特拉津污染的土壤稀释后,涂布于加有土壤浸出液和阿特拉津农药的平板,分别从两个采自不同地区的污染土壤中各分离了一株高效广谱降解菌AG1和ADG1：它们能以阿特拉津为唯一碳源、氮源和能源生长,能分别在44h和48 h内降解1 000mg L^-1的阿特拉津,降解率100%;它们还能以扑草净、西玛津等三嗪类除草剂为唯一氮源生长.16S rDNA核苷酸序列分析结果表明菌株AG1与ADG1都与节杆菌属（Arthrobacter）的细菌有高度同源性,结合两株菌的形态特征及生理生化特征,将它们鉴定为Arthrobacter spp..PCR扩增两株菌的降解基因,结果表明它们的降解基因都是trzN和atzBC的组合,这是国内首次报道具有该基因类型的阿特拉津降解菌.     

联苯菊酯降解菌的筛选、鉴定及降解特性研究

联苯菊酯是一种广谱高效杀虫剂,大规模的应用使其广泛残留在环境中,因此筛选联苯菊酯的高效降解菌具有重要意义。从扬州农药厂附近的地表土壤取样,利用富集驯化培养分离得到一株编号为S8的降解细菌,经表形特征、生理生化特性和16S rDNA序列分析其为醋酸钙不动杆菌（Acinetobacter calcoaceticus）,该菌株在pH7.0和30 ℃的条件下,对100 mg·L-1联苯菊酯的3 d降解率达56.4%,半衰期为60.7 h。其最适生长条件为：pH6.0~8.0,温度30~35 ℃,接种量5%。研究结果可为今后治理联苯菊酯残留污染提供理论参考。     

一株敌敌畏降解菌DDV-1(Ochrobactrium sp.)的分离鉴定及其降解特性的研究

Soil organic carbon （C） and total nitrogen （N） pools of a Chinese fir （Cunninghamia lanceolata （Lamb.） Hook.） （CF） forest, and an evergreen broadleaf （EB） forest located in mid-subtropical, southeastern China, were compared before clearcutting, with the effect of slash burning on organic C and total N in the top 10 cm of soil before and after burning also being evaluated. Prior to clearcutting CF forest had significantly lower （P 〈0.05） organic C and total N in the soil （0-100 cm） compared to EB forest with approximately 60% of the C and N at the two forest sites stored at the 0 to 40 cm soil. In post-burn samples of the 0-10 cm depth at 5 days, 1 year, and 5 years for CF and EB forests, significantly lower levels （P 〈0.05） of organic C and total N than those in the pre-burn samples were observed. Compared to the pre-burn levels, at post-burn year 5, surface soil organic C storage was only 85% in CF forest and 72% in EB forest, while total N storage was 77% for CF forest and 73% for EB forest. Slash burning caused marked long-term changes in surface soil C and N in the two forest types.     

采用Biolog, DGGE 和PLFA三种方法研究利用方式对土壤微生物群落结构的影响

Biolog, 16S rRNA gene denaturing gradient gel electrophoresis （DGGE）, and phospholipid fatty acid （PLFA） analyses were used to assess soil microbial community characteristics in a chronosequence of tea garden systems （8-, 50-, and 90- year-old tea gardens）, an adjacent wasteland, and a 90-year-old forest. Biolog analysis showed that the average well color development （AWCD） of all carbon sources and the functional diversity based on the Shannon index decreased （P 〈 0.05） in the following order： wasteland 〉 forest 〉 tea garden. For the DCCE analysis, the genetic diversity based on the Shannon index was significantly lower in the tea garden soils than in the wasteland. However, compared to the 90-year-old forest, the tea garden soils showed significantly higher genetic diversity. PLFA analysis showed that the ratio of Gram positive bacteria to Cram negative bacteria was significantly higher in the tea garden soils than in the wasteland, and the highest value was found in the 90-year-old forest. Both the fungal PLFA and the ratio of fungi to bacteria were significantly higher in the three tea garden soils than in the wasteland and forest, indicating that fungal PLFA was significantly affected by land-use change. Based on cluster analysis of the soil microbial community structure, all three analytical methods showed that land-use change had a greater effect on soil microbial community structure than tea garden age.     

Biodegradation of carbofuran by <Emphasis Type="Italic">Pichia anomala</Emphasis> strain HQ-C-01 and its application for bioremediation of contaminated soils

A novel yeast named HQ-C-01 was isolated from activated sludge and identified as Pichia anomala based on the morphology and 18S rDNA sequence analysis. The HQ-C-01 strain degraded 95.2% of carbofuran when the insecticide was used as the only C source and added at 50 mg/L in a mineral salts medium within 48 h. The optimal concentration, temperature, and pH of medium for degradation of carbofuran were 50 mg/L, 30°C, and pH 7.5, respectively. Strain HQ-C-01 could also effectively degrade other carbamate insecticides including carbaryl, indoxacarb, and fenobucarb, and the degradation rates were 99%, 85%, and 67%, respectively. Gas chromatography–mass spectrometry analysis showed that the strain metabolized carbofuran to produce benzofuranol as the intermediate metabolite, which was further degraded. Degradation of carbofuran added at 50 mg/kg of soil was higher in yeast-inoculated soil than in the control. These results indicated that strain HQ-C-01 may potentially be used in bioremediation of carbofuran-contaminated soil.     

猪毛角蛋白降解菌的分离筛选及其降解特性研究

为了筛选能高效降解猪毛角蛋白的微生物,该研究利用猪毛角蛋白为唯一碳氮源的培养基从土壤中分离角蛋白降解菌,并对其菌种分类及降解角蛋白的特性进行了研究。通过驯化共筛选到6株能够降解猪毛角蛋白的菌株,经过7 d发酵培养,菌株X-3对猪毛角蛋白的降解率达77.3%,降解效果最佳。结合菌株形态特征、生理生化特征及16S rDNA发育进化树分析,初步鉴定该菌株为凝结芽孢杆菌(Bacillus coagulans)。菌株X-3是一株广谱的角蛋白降解菌,对猪毛、羊毛、鸡毛、鹅毛、鸭毛的降解率均大于60%。该菌株在角蛋白底物作用下可产生角蛋白酶和二硫键还原酶,且酶活与猪毛角蛋白降解率变化存在显著的正相关性,表明菌株X-3产生的角蛋白酶和二硫键还原酶在角蛋白降解中起到非常关键的作用。在降解过程中,产生了大量的可溶性蛋白,二硫键中的硫也随之转化为硫酸盐、亚硫酸盐和巯基化合物,其中硫酸盐是主要的转化形式。该菌株的分离筛选丰富了猪毛角蛋白降解菌的微生物资源库,在氨基酸饲料生产中具有潜在的应用价值。     

一株高浓度烟碱降解菌的筛选、分离和初步鉴定

采用以烟碱为唯一碳源氮源的选择培养基,从烟草和植烟土壤中分离筛选出15株高浓度烟碱降解的菌株,其中菌株D9烟碱降解能力最强,其烟碱降解率为82％,耐受烟碱的最高浓度为8g／L。经常规形态特征、16SrDNA同源序列以及系统进化树分析,初步鉴定该菌株为类似氧化微杆菌,命名为Microbacteriumsp．GYC29。本研究为微生物降解烟碱的研究及应用提供了菌种资源。     

一株芽孢杆菌属的苄嘧磺隆降解菌的分离鉴定和降解研究

The objectives of this study were to isolate a bensulfuron-methyl (BSM)-degrading strain of Bacillus spp. and to evaluate its effectiveness in remediation of a BSM-contaminated soil. A BSM-degrading bacterium, strain L1, was successfully isolated in this study. Strain L1 was identified as Bacillus megaterium based on its morphological, physiological, and biochemical properties, G+C content, phylogenetic similarity of 16S rDNA, and fatty acid compositions. Two experiments were used to examine BSM degradation by strain L1. When BSM was used as a sole carbon source in a mineral salt medium, the average degradation rate of BSM by strain L1 was 12.8%, which suggested that the strain was able to utilize BSM as a sole carbon and energy source. In addition, supplement of yeast extract (200 mg L-1) significantly (P ≤ 0.01) accelerated the degradation of BSM by strain L1. Almost complete degradation (97.7%) of BSM could be achieved in 84 h with addition of yeast extract. In addition, when a sterile soil was supplemented with BSM (50 mg L-l), BSM degradation rate was 94.3% in 42 d, indicating the potential of using microbes for the remediation of BSM-contaminated soils in fields.     

氡温泉中耐辐照嗜热菌的分离及其特性研究

氡温泉水样经8kGy60Co γ射线预处理后,从中分离获得1株耐60Co γ射线和UV辐照的嗜热菌株R4-33.经形态观察、生理生化试验、脂肪酸分析、(G+C)mol%含量和16S rDNA序列测定,结果表明,菌株R4-33为杆状,革兰氏阴性菌,无鞭毛,形成末端芽孢;最适生长温度60℃,最适生长pH 7.5;能以葡萄糖...