2021年新乡市一起山夫登堡沙门菌食物中毒分离株的病原特征分析

目的 分析2021年新乡市一起山夫登堡沙门菌食物中毒分离株的病原特征。方法 对食物中毒事件进行流行病学调查;对采集的11份样本进行致病菌分离鉴定;对分离出的10株沙门菌进行血清学分型、药敏试验、5种毒力岛（SPIs）特征基因片段（mogA、sseL、mgtC、bcfA、araB）检测及脉冲场凝胶电泳（PFGE）分型分析。结果 10株沙门菌血清抗原式均为1,3,19;g,s,t,即山夫登堡沙门菌。10株沙门菌对头孢唑啉、卡那霉素、庆大霉素、阿米卡星的耐药率为100%,其中从患者粪便中分离出的2株对氨苄西林、四环素、多西环素、氯霉素、复方新诺明的耐药率为100%。PFGE图谱聚类分析显示,10株菌（2株病例株,5株食品株,3株环境株）之间条带无差异,高度同源。10株菌中5种毒力岛特征基因片段均检出。结论 本起食物中毒事件致病因子为山夫登堡沙门菌,该菌携带5种毒力岛特征基因片段,2株病例株沙门菌具有多重耐药性,建议相关部门高度关注。     

Identification of the glycosylation pattern of meat proteins in tissues from bovine myelin

A variant form of Creutzfeld–Jacob disease in humans is associated with the consumption of food contaminated with the bovine central neural system. This has focused attention on the need for procedures to detect tissues of the neural system in meat and meat products. A method was developed for the identification of myelin glycoproteins in bovine neural tissue. The glycosylated structures of glycoproteins in different protein mixtures from central nervous system (CNS) and peripheral nervous system (PNS) isolated from bovine myelin were identified by the specific lectin–glycoprotein reactions. Digoxigenin-labeled lectins bonded to the terminal glycoconjugate sequences of nine CNS glycoproteins ranging from 15 to 200 kDa and four PNS glycoproteins ranging from 22 to 105 kDa. Datura stramonium (DSA) recognized the epitope Galβ1–4GlcNAc by two CNS and three PNS glycoproteins. Maackia amurensis (MAA) recognized the epitope NeuAcα2–3Gal by four CNS and two PNS glycoproteins. The peanut Arachis hypogaea (PNA) reacted with the high molecular CNS glycoprotein (200 kDa) with the sequence Galβ1–3GalNAc. Galanthus nivalis (GNA) bonded to mannose subunits linked α1–3 and α1–6 by six CNS glycoproteins with molecular weights between 17 and 200 kDa. Four of these glycoproteins were recognized from monoclonal antibodies against the “HNK-1 epitope”.     

Improvement in the remaining activity of freeze-dried xanthine oxidase with the addition of a disaccharide–polymer mixture

In order to improve the remaining activity of a practically important freeze-dried enzyme, xanthine oxidase (XOD), the effects of disaccharide (sucrose and trehalose), polymer (bovine serum albumin: BSA and dextran) and a mixture of them on the loss of XOD activity during freeze-drying and subsequent storage were investigated. All samples were amorphous solids and their glass transition temperatures (T_g) were evaluated by using differential scanning calorimetry. Although dextran showed no stabilizing effect on the freeze-dried XOD, the others protected XOD from the activity loss during freeze-drying to a certain extent. It was found that the mixture of disaccharide (sucrose or trehalose) and BSA improved the XOD activity synergistically. The XOD activity of the samples decreased gradually during storage at a temperature range of between 25 and 60 °C. Samples stored at temperatures below the T_g showed a lower loss of XOD activity than those stored at just the T_g.     

Crinumin,a chymotrypsin-like but glycosylated serine protease from Crinum asiaticum: Purification and physicochemical characterisation

Plant latex could be a potential source of novel proteases usable in the food and feed industries because of broad substrate specificity with high stability in extreme conditions. Crinumin, a glycosylated serine protease with chymotrypsin-like activity was purified from the latex of Crinumasiaticum using cation-exchange column chromatography. Crinumin shows activity over a wide range of pH (4.5–11.5 and optimum at 8.5), temperature (75 °C and optimum at 70 °C) and is also functional against chaotrophs, organic solvents, and detergents, even after prolonged exposure. The molecular mass (67.7 kDa), extinction coefficient (17.7), isoelectric point (6.9), and numbers of tryptophan (13), tyrosine (24) and cysteine (15 with 7 disulphide bridges) residues were estimated. K_m of the enzyme was 31.7 μM with casein and 5 × 10⁴ μM with N-succinyl-l-phenylalanine-p-nitroanilide. Easy availability of the aqueous latex, simple purification procedure, high yield (33%), stability and activity in adverse conditions makes it applicable for the pharmaceutical and food industries.     

Effects of sulfated fucan,ascophyllan, from the brown Alga Ascophyllum nodosum on various cell lines: A comparative study on ascophyllan and fucoidan

The effects of fucose-containing sulfated polysaccharides, ascophyllan and fucoidan, isolated from the brown alga Ascophyllum nodosum, on the growth of various cell lines (MDCK, Vero, PtK₁, CHO, HeLa, and XC) were investigated. In a colony formation assay, ascophyllan and fucoidan showed potent cytotoxic effects on Vero and XC cells, while other cell lines were relatively resistant to these polysaccharides. Almost no significant effects of these polysaccharides were observed in the cell lines tested using the Alamar blue cytotoxicity assay over 48 h with varying initial cell densities (2500–20,000 cells/well) in growth medium. Interestingly, a significant growth promoting effect of ascophyllan on MDCK cells was observed, whereas treatment with fucoidan showed growth suppressive effects on this cell line under the same experimental conditions. These results suggest that ascophyllan is distinguishable from fucoidan in terms of their bioactivities. This is the first report of the growth promoting effects of a sulfated fucan on a mammalian cell line under normal growth conditions.     

乳化剂替代部分油脂对黄鸡生产性能、屠宰性能及血清相关指标的影响

将288只黄鸡随机分为3个组,分别饲喂3种日粮:基础日粮、"基础日粮+乳化剂(0.5 kg/t)-0.5%油脂+0.45%玉米"和"基础日粮+乳化剂(1 kg/t)-1%油脂+0.9%玉米",研究添加不同水平乳化剂替代油脂对黄鸡生产性能、免疫器官指数、屠宰性能及血清相关指标的影响。结果表明:添加乳化剂同时减少油脂不影响黄鸡的生产性能,经济效益有所改善。对器官指数和屠宰性能无明显影响,改善了部分胴体指标;极显著降低了肝脏中甘油三酯的含量。     

人血清白蛋白对变色酸2R的退色光谱性质及其分析应用

在pH为2.60的B-R缓冲介质中,变色酸2R与人血清白蛋白形成复合物,导致变色酸2R退色,吸光度的降低与人血清白蛋白的浓度成正比.线性范围为0～60 μg/mL,灵敏度ε=5.53×105 L·mol-1·cm-1.用于人血清白蛋白的测定,方法稳定,手续简便,结果满意.     

人脑钠肽与人血清白蛋白融合蛋白在毕赤酵母中的分泌表达及其活性

目的在毕赤酵母中表达重组人脑钠肽(BNP)与人血清白蛋白(HSA)融合蛋白,并检测其活性。方法重叠PCR法拼接BNP二联体与HSA基因,插入表达载体pPIC9K,电穿孔法转化毕赤酵母GS115,甲醇诱导表达,并对表达产物进行SDS-PAGE分析、Western blot鉴定及活性检测。结果融合基因(BNP)2-HSA经测序正确,重组表达质粒经双酶切鉴定构建正确。诱导72h,融合蛋白表达量最高,可达200mg/ml,且具有良好的反应原性,其活性为rhBNP标准品的1%。结论已在毕赤酵母中成功表达了具有一定生物活性的(BNP)2-HSA融合蛋白,为开发BNP长效药物奠定了基础。     

H5N1亚型禽流感病毒免疫血清精制工艺的建立

目的建立马抗H5N1亚型禽流感病毒免疫血清的精制工艺。方法以硫酸铵盐析法提取免疫马血浆中的IgG抗体,以8～256μg/mgIgG胃蛋白酶(活性单位1:3000)进行消化,以阳离子交换层析柱纯化F(ab’)2抗体,并参照《中国药典》三部(2005版)要求,对试制的样品进行检定。结果经一步50%硫酸铵和多步33%硫酸铵盐析,获得了较为纯净的IgG抗体。8μg胃蛋白酶用量可完全消化1mgIgG抗体分子,阳离子交换方法分离F(ab’)2抗体纯度可达90%以上,高于常规工艺制备的抗体纯度。以此工艺试制的样品,各项质量指标均符合《中国药典》三部(2005版)质量标准。结论已初步建立了马抗H5N1亚型禽流感病毒免疫血清的精制工艺。