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.

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.

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.

试验确定了由实验室保藏的沼泽红假单胞菌R1菌株适宜的发酵培养条件,为其作为饲料添加剂生产应用打下基础.采用正交试验L16(45)的设计对发酵培养基配方进行优化,采用单因素随机试验对培养条件进行优化.试验确定的发酵培养基配方为:甘油1.0 g、氯化铵1.0 g、磷酸氢二钾0.3 g、微量元素溶液0.05 ml、维生素溶液0.15 ml、蒸馏水1000 ml;最佳培养条件为:初始pH值7.0、培养温度30℃、接种量6％、装液量为液体培养基50 ml/250 ml三角瓶,40 W白炽灯下距离20 cm处照射静置培养.在此条件下,发酵培养7d,沼泽红假单胞菌R1菌数OD660值可达2.066,沼泽红假单胞菌R1菌数可达5.6×108个/ml.

介绍了无棒代料栽培天麻新技术，这是一种低成本、高收益的栽培模式，在天麻栽培中具有很重要的现实意义。

The nattokinase crudes were prepared and drying with different methods,and the influence of drying temperature and treating times on the nattokinase crudes were tested.It was found that the activity of nattokinase and electrophoretic spectrometry of different strains were different from each other.Both of freeze-drying and static microwave vacuum drying methods were suitable for nattokinase drying.The during time and the temperature(50 ℃ or above)have a serious influence on nattokinase activity of the crudes,and have 80% activity for 24 monthes preservation under the room conditions.

The fermentation technology of β-glucosidase by Absidia corymbifera in lab was studied. In order to improve the enzyme activity of the strain,the dynamic changes of the enzyme production fermentation process was researched. Compositions of medium and fermentation conditions of the D8 strain were optimized by single factor test. The optimum medium compositions:corn starch 5%;the urea 0.5%;added inorganic salt CaCl2,MgSO4·7H2O,KH2PO4,FeSO4·7H2O;medium volume 120 mL/250 mL,initial pH 3.0,the best temperature 25 ℃,the time course8 d. In this way,it could produce β-glucosidase activity 168.7 U/mL in 7 d.

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.

To establish the TC rapid detection strip with colloidal gold labeling for detecting on tetracycline(TC) residue in food,coating material is made of nitrocellulose filters,colloidal gold labeling monoclonal antibody against tetracycline(TC mAb)is made from the lab by ourselves.Colloidal gold labeling TC mAb collosol is 40% of its original.Test line(T line)is coated with TC-BSA of 1.5 mg/mL,while Control line(C line)is coated with rabbit anti-rat IgG of 2.5 mg/mL.This test strip sensitivity is 50 ng/mL.The novel method coincidence rate is 100% compared with ELISA in 100 milk samples.This strip is suitable for the TC rapid detection in food animal farm,food processing plant,food entry-exit inspection and quarantine system.

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.

To establish a rapid,simple colloidal gold immunochromatographic assay(GICA) for the detection of chloramphenicol(CAP)residues.Nanocolloidal gold particles were prepared and labeled to an anti-CAP monoc-lonal antibody(MAb).CAP was conjugated to bovine serum albumin(BSA) and dispersed on a nitrocellulose(NC) membrane to be the test line(T).The more analyte present in the sample,the more effectively it would compete with the CAP-BSA for binding to the limited amount of gold labeled anti-CAP MAb.The presence or absence of a colored band on the test line indicates a negative or positive result.A total of 60 milk samples were detected for CAP by GICA and ELISA.The corresponding rate of both tests was 100 %.GICA is a sensitive,specific,simple and rapid assay for detecting CAP in the sample.

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.

根据辣椒辣味成分和色素成分理化性质的不同,经过多次不同浓度、不同酸碱度溶剂的提取分离试验研究,确定了简便易行、安全有效地分离两类成分的技术工艺.

胭脂豆红色素来源于蔬菜,可溶于水,染色力强,呈色艳丽.但是,天然色素普遍存在稳定性差的问题.为了更好地使用该色素,文中在不同条件下进行了稳定性试验,基本摸清了该色素的稳定特性,并就具体情况提出了不同的注意事项.