通过紫外线诱变植物乳杆菌选育具有良好发酵性能的液体发酵饲料专用乳酸菌.试验中采用紫外线的最佳诱变时间为120 s,并用溶钙圈法对诱变后的菌株进行初步筛选,在MRS-CaCO3培养基上得到溶钙圈直径较大的菌株79株.通过对这79株菌株的液体发酵饲料进行乳酸含量的测定,最终得到一株乳酸高产菌株UR-3,显著提高了产乳酸量(p<0.05),在液体发酵饲料中,该菌株产乳酸能力较出发菌株提高了44.53％.

制备抗沙门氏菌O8因子单克隆抗体,建立C2、C3亚群沙门氏菌酶联免疫吸附分析检测方法.用加热灭活的纽波特沙门氏菌免疫BALB/c小鼠,通过细胞融合建立分泌抗体的杂交瘤细胞株.用ELISA法检测及鉴定单克隆抗体,酶联免疫吸附分析法初步判断抗体应用于检测试剂的可能性.得到4株与沙门氏菌C2、C3亚群菌株发生反应的单克隆抗体细胞株,分别命名为6F2、7A2、8D8和8D9;4株抗体与肠道正常细菌和常见的致病细菌无交叉反应;单克隆抗体免疫球蛋白类型分别为小鼠IgG1、IgG3、IgG2b、IgM,轻链均为kappa;用单克隆抗体7A2和8D8研制的酶联免疫吸附分析试剂盒,对沙门氏菌C2亚群的纽波特沙门氏菌、波那雷恩沙门氏菌和C3亚群的肯塔基沙门氏菌的最低检出值均可达到105 cfu/mL.获得的单克隆抗体具有较高的特异性,有可能应用于生产检测C2和C3亚群沙门氏菌的酶联免疫吸附分析试剂盒.

制备抗沙门氏菌O4抗原单克隆抗体并鉴定其特性.用加热灭活的乙型副伤寒沙门氏菌免疫BALB/c小鼠,通过细胞融合方法建立分泌抗体的杂交瘤细胞株.用包被沙门氏菌全菌和纯化的乙型副伤寒沙门氏菌LPS的间接ELISA法筛选杂交瘤细胞和鉴定抗体.其中3株杂交瘤细胞分泌的抗体初步判断为针对O4抗原的抗体,分别命名为1E12、2D5和4C2,这3株抗体与肠道正常细菌及常见的致病性细菌无交叉反应;单克隆抗体的免疫球蛋白类型分别为IgG1κ和IgMκ.获得的单克隆抗体具有较高的特异性,有可能用于B群沙门氏菌免疫学检测方法的建立.

The objective developed a rapid and simple colloidal gold immunochromatographic assay(GICA)for the detection of cTnI.A colloidal gold immunochromatography test based on the double-antibody sandwich assay for detecting cTnI was developed. Its sensitivity,specificity and stability were evaluated. The results showed the typical detecting time was not more than 10 min per sample. The positive sample produced a positive signal on immunochromatographic strip,whereas the negative one did not.The sensitivity of the test was 1.0 ng/mL.The positive coincidence rate was 97.0% and the negative coincidence rate was 100.0%.The overall coincidence rate between two assays was 98%.The performance of gold-immunochromatography test strip was stable.

A rapid test on procalcitonin,a new inflammatory marker,was researched by colloidal gold immunochromatography assay during priliminary experimental period.Sandwich method was adopted in the assay.Its test line was coated with lab homemade PCT-F12 McAb,and its control line was lab homemade rabbit anti-mice IgG.Its colloidal gold pad was coated with lab homemade colloidal gold of labeled PCT-D7 McAb complex.The results were judged by the color reaction in the strips.And 52 clinical serum samples were tested by immunochromatography assay and immunochemiluminometric assay.The overall coincidence rate between two assays was 95％.The reference sensibility of the procalcitonin strip which established tentatively was 0.05 ng/mL.Both of the precision and the stability were good.

BALB/c mice were immunized with artificial immunogen SAL-BSA. Three hybridoma lines secreting antibody against salbutamol were established by the cell fusion technology. The biological characteristics of momoclonal antibodies were identified. The rusults showed that the 50% inhibitiong concentrition of free SAL for these monoclonal antibodies were 1.41,5.53 and 9.62 ng/mL respectively. The cross reaction rate of 2F7 with clenbuterol, mabuterol,terbutaline were 217%,128%and 70.5%respectively. It was not found that the cross-reaction of 2F7 with ractopamine,epinephrine,norepinephrine,and cimaterol. The antibodies from the three hybridoma lines were mouse IgG1. The sensibility of colloidal gold immune chromatography strip were prepared by mAb 2F7 was 3 ng/mL for SAL standrand. Therefore,the mAb 2F7 might be used to establish the immunology detection mothed for SAL residual.

目的 建立人尿液中铅含量的目视比色快速测定方法.方法 在表面活性剂增溶作用下,尿液中的铅离子在酸性介质中与硫氰酸盐和甲基紫形成三元离子缔合物,该胶束增溶体系随三元缔合物胶束的浓度不同而显示不同的颜色,以目视比色法进行半定量分析.结果 硫氰酸钠、硫酸、甲基紫、OP乳化剂的浓度分别为1.50～2.50 mol/L、0.04～0.45 mol/L、0.5～30.0 mmol/L、12.4～18.5 mmol/L时显色效果最理想.以胶束增溶法测定120份尿液样本,同时与双硫腙比色法(SP法)测定结果比较,两种结果的符合率为99.17％.结论 该方法操作简便快速,结果准确直观,可用于人尿液铅含量的半定量快速测定,适用于铅污染人群的普查.

目的 筛选并鉴定软腐白菜根系土壤中分离出的细菌.方法 从河南省大白菜田成熟白菜病株根系土壤采集样品,利用模拟原环境的培养基成分和条件,对样品中的细菌进行分离和培养.对菌株进行形态学观察、生理生化特征鉴定以及16S rRNA基因序列测定、比对分析和系统发育树构建,对菌株进行初步分类鉴定.结果 分离到的一株编号为Q2S2-18A2的菌株16SrDNA与代夫特菌属内的Delftia lacustris 332T(EU888303)具有最高的相似性,相似值为98.60％.结论 模拟原环境培养基,结合经典分类学方法和16S rRNA基因序列比对分析、构建系统发育树,可分离并鉴定软腐白菜根系土壤中的机会致病菌湖生代夫特菌.

目的 对采自河南省临颍县麦田的土壤样品进行苯磺隆耐受菌的筛选和初步鉴定.方法 利用选择性培养基,经富集培养,从长期施用苯磺隆的麦田土壤中筛选苯磺隆耐受菌.采用基于16S rRNA基因序列的系统发育分析和生物学特性研究,对菌株进行初步分类鉴定.结果 共分离到耐受苯磺隆的纯菌12株,其中一株LYBRD3-3对苯磺隆耐受能力最强,苯磺隆耐受浓度为150 μg/L.经过对该菌形态观察、生理生化特征及基于16S rRNA基因序列的系统发育分析表明,LYBRD3-3与无色菌属的Achromobacter insolitus LMG 6003T (AY170847)序列具有最高的相似性,同源性为99.58％,初步鉴定该菌为无色菌属菌.结论 分离自麦田土壤的苯磺隆耐受菌为进一步筛选苯磺隆降解功能菌株提供菌种资源,进而为污染土壤的原位生物修复提供基础研究.

Objective To establish a rapid method for semiquantitative determination of zinc in human urine by visual colorimetry.Methods In the presence of surfactant,zinc reacts with thiocyanate and crystal violet to form a ternary ion-association complex,which can show different colors with the concentration change.Results This method was compared with routine pyridylazo naphthol method(PAN) by testing 36 urine samples and the coincidence rate was 97.22%.Conclusion The method is simple,rapid,accurate and intuitive,which can be used for semiquantitative determination of zinc content in urine.

To isolate and identify Empedobacter brevis strains in rhizospheric soil of Chinese cabbage soft rot,15 aroma bacterial strains were isolated with selected medium,from the rhizospheric soil where the Chinese cabbages were planted.The isolated strains were identified according to their 16S rRNA gene sequences analysis as well as their morphological features,physiological and biochemical analysis.Their taxonomic statuses were confirmed by their 16S rRNA gene sequences homologous analyses and phylogenetic trees.The result showed that 12 aroma strains exhibited the highest levels(99.86%) of similarity to Empedobacter brevis LMG 4011T(AM177497).

The industrial production of β-glucanase was studied in the solid-state fermentation,by Aspergillus niger An08-752 strain with three kinds of equipments.The result showed:the β-glucanase activity reached 1.15× 105 μ/g dry koji at 30～32 h by thick layer ventilation koji tank in the solid-state fermentation;the β-glucanase activity reached 1.28×105 μ/g dry koji at 35～37 h by fermentation tank in the solid-state fermentation;the βglucanase activity reached 1.36×105 μ/g dry koji at 32～34 h by koji tray in the solid-state fermentation.

A strain of genus Pseudomonas, LYBRD3-7T was isolated from long-term sulfonylurea herbicides applied wheat-field soil in Linying located in Henan province of China. This strain is a strictly aerobic and Gram-negative short rod-shaped bacterium with single flagellum. Phylogenetic evaluation based on 16S rRNA gene sequence analysis placed this isolate as a member of Pseudomonas, and most closely to Pseudomonas tuomuerensis CGMCC 1.1365T (97.1 %) and P. alcaligenes IAM12411T (97.1 %). Morphological characters and chemotaxonomic data confirmed the affiliation of strain LYBRD3-7T to the genus Pseudomonas. The results of phylogenetic analysis, physiological and biochemical studies, and DNA–DNA hybridization allowed the differentiation of genotype and phenotype between strain LYBRD3-7T and the phylogenetic closest species with valid names. The name proposed for the new species is Pseudomonas linyingensis sp. nov. The type strain is LYBRD3-7T (=CGMCC 1.10701T  =LMG 25967T).

A high temperature-resistant bacteria strain P98-18A1 capable of growing in petroleum hydrocarbons at 55℃was isolated from hot produced fluid from Zhongyuan oilfield.This strain was identified as Bacillus sp.based on phenotypio,physiological,biochemical and 16S rDNA sequence analysis.With crude oil and several alkanes degradation experiments,results showed that this strain can effectively utilize C_6- C_(16) n-alkanes,and selectively utilize of the chain longer than C_(16) alkanes to some extent under the optimum conditions.

A Gram-negative, non-mobile, polar single flagellum, rod-shaped bacterium WZBFD3-5A2 T was isolated from a wheat soil subjected to herbicides for several years. Cells of strain WZBFD3-5A2 T grow optimally on Luria-Bertani agar medium at 30 °C in the presence of 0–4.0 % (w/v) NaCl and pH 8.0. 16S rRNA gene sequence analysis revealed that strain WZBFD3-5A2 T belongs to the genus Pseudomonas . Physiological and biochemical tests supported the phylogenetic affiliation. Strain WZBFD3-5A2 T is closely related to Pseudomonas nitroreducens IAM1439 T , sharing 99.7 % sequence similarity. DNA–DNA hybridization experiments between the two strains showed only moderate reassociation similarity (33.92 ± 1.0 %). The DNA G+C content is 62.0 mol%. The predominant respiratory quinine is Q-9. The major cellular fatty acids present are C 16:0 (28.55 %), C 16:1ω6c or C 16:1ω7c (20.94 %), C 18:1ω7c (17.21 %) and C 18:0 (13.73 %). The isolate is distinguishable from other related members of the genus Pseudomonas on the basis of phenotypic and biochemical characteristics. From the genotypic, chemotaxonomic and phenotypic data, it is evident that strain WZBFD3-5A2 T represents a novel species of the genus Pseudomonas , for which the name Pseudomonas nitritereducens sp. nov. is proposed. The type strain is WZBFD3-5A2 T (=CGMCC 1.10702 T = LMG 25966 T ).

Objective To isolate and identify the conditioned pathogenic bacteria strain Myroides odoratus.Methods With selected medium,a conditioned pathogenic bacteria strain was isolated from the rhizospheric soil where the Chinese cabbages were planted.The isolated strain was identified according to its 16S rRNA gene sequence analysis as well as its morphological features,physiological and biochemical analysis.Its taxonomic status was confirmed by 16S rRNA gene sequence homologous analysis and phylogenetic tree.Results The standard strain exhibited the highest levels(99.99%) of similarity to the Myroides odoratus M58777,the results of several methods and morphological observation,this strain was finally identified as Myroides odoratus.Conclusion There is a conditioned pathogenic microorganism,Myroides odoratus,in the rhizospheric soil of Chinese cabbages.

OBJECTIVE To investigate the diversity of bacteria in soft rot Chinese cabbage and analyze their correlation with rhizosphere bacteria, we analyzed the bacterial population structures of soft rot Chinese cabbage and the rhizosphere in different habitat. METHODS Based on the initial medium and artificial Chinese cabbage medium, we isolated the bacteria from soft rot tissues and rhizospheric soils from two typical habitats. According to the analysis of 16S rRNA gene sequence homology, we identified the isolated strains and analyzed the strains population structure. RESULTS The total bacteria in soft rot tissues were 4.0 x 10(8) cell g(-1) and 1.2 x 10(11) cell g(-1), the number of pure strains were 56 and 85, the dominant strains were Curtobacterium flaccumfaciens pv. flaccumfaciens and Pseudomonas spp. (P. hibiscicola, P. taiwanensis, P. tuomuerensis, P. mosselii). The total bacteria in rhizospheric soils were 2.7 x 10(5) cell g(-1) and 6.2 x 10(7) cell g(-1), the number of pure strains were 36 and 70, the dominant strains were Bacillus megatherium and Pseudomonas spp. (P. plecoglossicida, P. hibiscicola, P. parafulva, P. monteilii, P. geniculata). CONCLUSION The methods used in this study were effective in analyzing bacterial diversity in soft rot Chinese cabbage and the results correlated well with the soil bacteria analysis, suggesting that soft rot Chinese cabbage may be induced by various environmental bacteria. Our results infer that soft rot of Chinese cabbage might be pathogen-complex, and provide the clues for the mechanism study and protection.

From 17 strains of Aspergillus niger,we obtained five strains of A.niger by using the plate transparent circle method with oxford cup.A.niger niger-2 was selected according solid fermention experiments for enzyme production,the results showed it produced high β-glucosidase activity,higher cellulase activity(CMC) and the filter paper activity(FBA) of the strain during the same fermentation.β-glucosidase activity reached to 78.75 U /g,cellulase activity reached to 312.15 U /g,and the filter paper activity reached to 782.6 U/g.

A method is developed for the quantitative analysis of tribenuron-methyl by reversed-phase high performance liquid chromatography,in MS medium,on C18 Column with uv-absobance detector.The samples of ahead processing design adopt dichloromethane extraction and revolving evaporator and methanol dissolve.The linear relation was very good in the range of 25~200 μg/mL(R2=0.999).The samples standard deviation was 0.082,the coefficients of variation was 0.16 %,and the average recovery was 100.86 %.This method can accurately determinate tribenuron-methyl content in the medium.

Two novel actinomycetes, designated DQS3-9A1(T) and DQS3-9A2, were isolated from a saline soil contaminated with crude oil in the Shengli Oilfield in China. On the basis of 16S rRNA gene sequence analysis, the two strains were most closely related to Mycobacterium species (92.7-94.9 % similarities), and formed a distinct lineage in the suborder Corynebacterineae . In addition, the major sugars in the cell wall, arabinose and galactose, supported the affiliation of strain DQS3-9A1(T) with members of the family Mycobacteriaceae. However, strain DQS3-9A1(T) did not contain mycolic acids and MK-8 (85.5 %) was the major menaquinone for both isolates. The major cellular fatty acids for strain DQS3-9A1(T) were C(16 : 0) (20.5 %), 10-methyl C(17 : 0) (19.3 %), 10-methyl C(18 : 0) (16.1 %), summed feature 3 (11.4 %), C(15 : 0) (11.3 %), C(17 : 0) (5.0 %) and C(17 : 1)omega8c (5.0 %). The polar lipids of strain DQS3-9A1(T) consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, phosphatidylinositol and an unknown glucosamine-containing phospholipid. These chemotaxonomic data indicated that strain DQS3-9A1(T) differs from the present members of the suborder Corynebacterineae. Therefore, the creation of Amycolicicoccus subflavus gen. nov., sp. nov. is proposed, with DQS3-9A1(T) (=DSM 45089(T)=CGMCC 4.3532(T)) as the type strain.