多重集合的运算研究与应用

集合是现代数学中最基本的概念之一,并已深入到各种学科和技术领域中.而作为集合概念扩展的多重集合概念,却少有应用.通过对多重集合进行系统研究,提出了多重集合的包含、相等、多重幂集,以及多重集合补集等相关概念和运算,建立了多重集合完善的理论体系,并给出了多重集合在古典概率中的应用.为进一步研究多重集合在各种学科和技术领域中的应用打下了良好的基础.     

谈氯化钙注射液在兽医临床上的应用

<正>众所周知,氯化钙注射液的主要成分就是氯化钙,因为氯化钙具有促进骨骼和牙齿钙化形成、维持神经肌肉的正常兴奋性、促进血凝和消炎、抗过敏等多重功效[1],所以氯化钙注射液已成为兽医临床上治疗各种疾病的常用药。但是,目前仍有许多兽医对本品的应用存在诸多误区,致使氯化钙的功效得不到     

仔猪致病性大肠杆菌的PCR快速检测

在对大肠杆菌主要毒力因子基因序列分析的基础上,建立了用于快速检测毒素、菌毛和毒力岛(HPI)的PCR方法;用所建立的多重PCR或单重PCR方法,对江苏部分地区临床疑似大肠杆菌感染的新生仔猪腹泻样品110份和健康新生仔猪粪便35份进行了快速检测。结果表明:HPI已经成为致仔猪腹泻大肠杆菌最常见的毒力因子(58.18%),其次是肠毒素(LT1 10.91%、ST2 8.18%、ST1 2.73%),并揭示了至少46.36%的病例与HPI+大肠杆菌感染相关,11.82%的病例与HPI+大肠杆菌和产肠毒素大肠杆菌(ETEC)混合感染相关,仅有6.36%的病例与ETEC感染相关。菌毛仍然是ETEC的主要毒力因子,其中987P+菌株6.36%、K88+菌株3.64%、K99+菌株0.91%,未发现F41+菌株。此外,通过对健康新生仔猪样品的检测,发现48.57%的动物携带HPI+大肠杆菌,5.71%的动物携带ETEC(主要为ST2+),未发现所检测的其他毒力因子,这表明HPI+大肠杆菌很可能是条件性致病菌。     

Isolation,Identification and Multidrug-resistant Analysis of the New Sequence Type ST989 Strain of Enterococcus faecium

Gram-positive cocci were isolated from half a month diseased lambs with a fatal infection in a sheep farm in Minhe county, Qinghai province.The clinical symptoms observed were drooping, reduced feed intake, emaciated and fever.The infected lambs died several hours after the disease been observed.Necropsy of two dying lambs revealed that for one lamb, the liver was enlarged and capsule of the liver dropped, and other organs had no obvious changes;For the other one, the organs had no obvious changes.After gram staining, gram-positive cocci were observed from all the samples.Partial Tuf gene sequences of these isolates were amplified and sequenced for species identification, BLASTn conparison result revealed that they had the highest sequence similarity with that of Enterococcus faecium (E.faecium) AUS0085 (99.8%), we thus primarily confirmed that these strains belonged to E.faecium.Antimicrobial susceptibility test indicated that these strains could be divided into two classes.Class 1 was resistant to β-lactams, aminoglycosides, macrolides, tetracyclines, quinolones, rifamycins, lincomycin and clindamycin;Susceptible to chloromycetin and vancomycin.Class 2 was resistant to β-lactams, aminoglycosides, tetracyclines, quinolones, rifamycins;Intermediate resistance to erythromycin and roxithromycin;But susceptible to clarithromycin, azithromycin, clindamycin, lincomycin, chloromycetin and vancomycin.Multilocus sequence typing (MLST) based on seven housekeeping genes for E.faecium revealed that E.faecium lby1 strain in class 1 and E.faecium lbg3 strain in class 2 were defined as a new sequence type ST989.Phylogenetic tree analysis revealed the representative strain of these isolates, E.faecium lbg3, was clustered together with E.faecium ST468.The pathogenicity test revealed that E.faecium lby1 and lbg3 strains both had pathogenicity.     

Study on Drug Resistance of Escherichia coli Isolated from Chicken and Swine in Organic Farm

To determine the drug resistance of E.coli strains from swine and chicken in organic farm,collection of stool specimens was conducted in one organic farm in Conghua city,Guangdong province in 2014,in which 118 E.coli strains were separated,including 55 from chicken and 63 from swine.The sensitivity of the E.coli strains from swine and chicken to 18 kinds of drugs was detected through agar dilution method.Similar properties in drug resistance were observed both in E.coli strains from swine and chicken in the same organic farm.In consideration of the resistance rate,samples were relatively sensitive to ceftazidime,cefquinome,cefoxitin,amikacin,apramycin,gentamicin,neomycin,florfenicol,chloramphenicol,imipenem,ciprofloxacin and olaquindox (chicken 0 to 21.8%;swine 0 to 14.3%).These isolates showed moderate resistance to ampicillin and streptomycin (chicken 29.1% to 38.2%;swine 23.8% to 27.0%).About 56.4% chicken isolates and 47.6% swine isolates showed resistant to more than 3 kinds of drugs.However,severe resistance was observed in trimethoprim-sulphamethoxazole,tetracyclineand and doxycycline (chicken 45.5%,76.4% and 72.7%;swine 50.8%,81.0% and 68.3%).The results also indicated that under organic farming mode,the drug resistance rates of E.coli strains from swine and chicken were relatively low in general.Multi-drug resistance existed but was relatively slight.Although the drug resistance rate to several specific antimicrobials was high,it was still lower than that in conventional farms.Since most studies focused on the drug resistance of the bacteria under conventional farming model,this study filled the blank of the research of drug resistance under organic farming model.     

玉米品种德美亚多重SSR-PCR系统的建立及应用

玉米杂交种纯度的优劣直接决定其产量,常规检测方法采用若干单引物进行若干次电泳检测,鉴定周期长,效率低,不能及时反映杂交种纯度,难以及时指导大田生产.为了提高玉米种子纯度检测的准确性,建立稳定高效的多重PCR检测技术,本研究以'德美亚1号'、'德美亚2号'、'德美亚3号'及其父母本为试验材料,通过比较不同DNA提取方法,...     

2种桑树病原真菌多重PCR检测方法的建立

目的 褐斑病与轮纹病是桑树Morus alba 的2种常见真菌性病害,危害严重,给生产上带来极大损失。本研究旨在快速准确地诊断2种病害的主要病原菌新褐斑壳丰孢Neophloeospora maculans与膝节霉Gonatophragmium triuniae。方法 基于多重PCR原理,针对2种病原菌的核糖体内转录间隔区(Internal transcribed spacer, ITS)序列设计多重PCR的特异性引物,优化多重PCR反应的条件,并通过对45份不同地区桑树褐斑病与轮纹病样本进行检测以验证所建立的多重PCR的可行性。结果 建立的多重PCR检测体系具有良好的可操作性,特异性良好,2种病原菌的DNA检测灵敏度分别达0.1 和1.0 pg/μL,通过对不同地区的田间收集的多份桑树病样进行检测,可以明显区分出不同地区2种病害病原菌的种类。结论 所建立的多重PCR技术可用于桑树褐斑病与轮纹病病原菌的快速检测,可为桑树真菌性病害的防控建立基础。     

不同环境胁迫因子对猪源耐药大肠杆菌生长的影响

【目的】探究环境胁迫因子对猪源多重耐药大肠杆菌生存适应性的影响,为创造多重耐药菌株带来高适应代价的环境及加速降低种群耐药水平提供理论依据。【方法】以分离自贵州省规模化猪场的多重耐药大肠杆菌QL15为研究对象,以大肠杆菌质控(敏感)菌株ATCC25922为对照,经体外竞争试验测定不同环境(培养基浓度、碳源、氮源、pH和温度)胁迫下耐药菌株QL15相对于敏感菌株ATCC25922的适应性,并通过重测序变异位点和实时荧光定量PCR测定分析耐药基因表型及外排泵和外膜孔道蛋白基因表达与细菌适应性代价的关系。【结果】耐药菌株QL15在体外独立及混合培养时,除在稀释LB培养基(1/4、1/8和1/16)中的长势较敏感菌株ATCC25922低外,在低碳源(0.1%和0.5%)、低氮源(0.01%和0.05%)、酸碱性(p H=5.0、pH=6.0和pH=8.0)及低温(20℃)和高温(42℃)胁迫下的生长均优于敏感菌株ATCC25922,且混合培养结果优于独立培养。将耐药菌株QL15基因组序列与敏感菌株ATCC25922基因组序列进行比对,共检测到509个InDel突变位点,其中Delete突变位点233个、Insert突变位点276个;共注释到71个功能基因,以细胞壁与细胞膜相关基因突变最多(10个),占14.1%。与敏感菌株ATCC25922相比,耐药菌株QL15在不同环境胁迫下其外排泵与外膜孔道蛋白相关基因emrB、acrB、tolC和ompC呈下调表达,而ompF基因呈上调表达。【结论】多重耐药大肠杆菌耐药性的获得促使其具有更强的适应性,但对可直接利用营养物质的获取弱于敏感菌株,即处于竞争劣势。耐药大肠杆菌的多重耐药性主要是由多种外排泵和外膜孔道蛋白介导完成,其中,ompF基因在pH胁迫、全营养胁迫和高、低温胁迫下均处于上调表达状态,故推测ompF基因的高表达有利于大肠杆菌在上述胁迫条件下生长。     

3种天然产物联合抗生素对肺炎克雷伯氏菌的协同抗菌作用

为筛选出联合抗生素对肺炎克雷伯氏菌具有协同抗菌作用的天然产物。采用药敏纸片法测定肺炎克雷伯氏菌对8种抗生素的耐药性,采用微量肉汤稀释法测定抗生素和3种天然产物的最小抑菌浓度,应用棋盘法分别测定花青素和咖啡酸与抗生素联用对肺炎克雷伯氏菌的抗菌活性,并通过生长曲线评估咖啡酸与抗生素联用的协同抑菌作用。结果表明,肺炎克雷伯氏菌对8种抗生素均耐药,抗生素的MIC值为16->512 μg/mL。花青素和咖啡酸具有较好的抗菌活性,MIC值分别为25 μg/mL和1875 μg/mL,肉桂醛无抗菌作用,MIC>250 μg/mL。花青素与头孢他啶和克林霉素联用对肺炎克雷伯氏菌具有协同抗菌作用,2种抗生素的MIC值降低了4倍,咖啡酸与8种抗生素存在协同作用,抗生素的MIC值降低了8~>4096倍。抗生素与咖啡酸联用的协同抗菌效果优于其与花青素的组合,且咖啡酸与磺胺甲恶唑和氯霉素联合使用能持续抑菌至少24 h。本研究首次报道了咖啡酸与抗生素联用对多重耐药肺炎克雷伯氏菌具有显著协同抗菌作用,表明联合疗法是消减细菌耐药性的一种有效策略。