This study was conducted to investigate the effect of Alfalfa polysaccharide (APS) on the growth performance and immune responses of broilers. A total of 400 1-day-old Avian broiler chicks were randomly divided into four treatment groups and administered intranasal with different levels of APS (0, 2.5, 5.0 or 10.0 mg/dose) on day 1 and day 14. Growth performance, phagocytic activity, T lymphocyte transformation, and serum antibody titers against the Newcastle disease virus (NDV) vaccine were examined. During the study period of 42 days, no significant differences were observed in food intake among treatment groups (P>0.05). The APS administered groups displayed a low FCR, especially the 2.5 mg groups (P<0.05). Broilers with 2.5 mg APS displayed a much higher phagocytic activity than the control group (P<0.05). At the same time, APS enhanced the T lymphocyte transformation in response to phytohaemagglutinin (PHA). The antibody titer to NDV was not influenced by APS on day 14, while it improved on day 28 and day 42, especially in the 2.5 mg group (P<0.05). And the above data suggest that APS could improve innate and cellular immune responses in broilers.

Porcine Parvovirus (PPV) and porcine circovirus type 2 (PCV2) often have mixed infection in the process of clinical breeding, and PCV2 infection will cause immunosuppression in pigs, which is easy to stimulate or complicated with other infectious pathogens. Haemophilus parasuis (HPS) is a typical `opportunistic" pathogen, which often leads to mixed infection with PCV2 as a secondary pathogen. In order to establish a rapid and simultaneous detection of three pathogens of PPV, PCV2 and HPS, referring to the relevant genome sequence of GenBank, specific primers were designed according to the conserved region of VP2 gene of PPV, Cap gene of PCV2 and infB gene of HPS, and the amplified fragments were cloned into the vector to construct plasmid standard. Using standard samples with different dilutions as templates, adjusting primer concentration, annealing temperature and other conditions, a real-time fluorescence quantitative PCR method for PPV, PCV2 and HPS triple SYBR Green I was established. Three specific Tm peaks could be generated on the same melting curve without cross-reaction with other pathogens. The minimum detection limits of this method were 153 copies/mu L, 128 copies/mu L and 91 copies/mu L, with good specificity and repeatability, which provided technical support for rapid diagnosis of these three diseases and could be used for clinical tissue material detection.

Actinobacillus pleuropneumoniae, Haemophilus parasuis, and Pasteurelle multocida are common pathogens of respiratory diseases in the pig industry, and they may cause secondary infections and serious economic losses to the pig industry. The clinical symptoms caused by these three pathogens are difficult to distinguish with the naked eye, and mix infections bring difficulties to the diagnosis of diseases. In this study, specific primers were designed on the basis of A. pleuropneumoniae Apx IV, H. parasuis Omp P2 and P. multocida PlpE gene. The expected amplified products ofA. pleuropneumoniae, H. parasuis, and P. multocida were 157, 120 and 305 bp, respectively. After the amplified fragment was cloned into a vector, a standard plasmid was constructed. By using the standard plasmid as template, a fluorescence quantitative PCR method for simultaneous detection ofA. pleuropneumoniae, H. parasuis, and P. multocida multiple SYBR Green I was established. Combined with melting curve analysis, the sensitivity, specificity, and repeatability were also evaluated. The results showed that the sensitivity of the method for detecting the three pathogens were 147, 145, and 61 copies/mu L. On the same melting curve that produced three specific Tm peaks, no cross reaction with other bacteria was observed, and the method demonstrated good specificity and repeatability. This method could be used for the simultaneous detection of the three pathogens, thus providing an effective detection tool for disease prevention and treatment.

Introduction:Porcine circovirus type 2 (PCV2) causes immune repression and intercurrent infections in pigs, resulting in a huge economic loss to the pig breeding industry. Additionally, the spread of PCV2 in pig farms can pollute the living environment of the residents in the farm's vicinity, which increases the rate of infections. Therefore, rapid and sensitive detection methods are needed for disease prevention and timely environmental cleaning.Methods:This research describes a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) that utilizes gold nanoparticles (AuNPs) in a functional, specific antibody labeled probe for the detection of PCV2. Due to their high specific surface area and histocompatibility, AuNPs were used as carriers of HRP labeled anti-PCV2 antibodies to amplify the detection signal.Results:Compared to conventional sandwich ELISA procedures, this method resulted in higher sensitivity (51-fold) and a shorter assay time with a limit of detection of 195 TCID50/mL. The cross-reactivity assay demonstrated that this assay was PCV2 specific.Conclusion:The amplified Ab (HRP) labeled AuNPs probe provides a sensitive analytical approach for the determination of the traces of the PCV2 antigen in early diagnosis.

In the present study, a duplex SYBR Green real-time PCR assay was developed in order to indentify Actinobacillus pleuropneumoniae and Haemophilus parasuis infection in one reaction, through a melting curve analysis. This method utilized two pairs of specific primers that allowed the amplification of highly conserved regions of A. pleuropneumoniae Apx IV and H. parasuis omp P2 gene. Reconstitution experiments were conducted by using PMD 19T plasmid in order to determine the sensitivity of the assay. The results showed that the Tm values of the melting curves of A. pleuropneumoniae and H. parasuis were 83.36±0.09oC and 76.48±0.17oC, respectively that could accurately distinguish these two pathogens. And no cross reaction were observed between other respiratory pathogens, which suggested a high specificity of two primers. The detection sensitivity of the assay was 127 and 96 copies/μL which was higher than that of the ordinary PCR detection methods. This rapid technique may present a simple, useful option for simultaneous detection of A. pleuropneumoniae and H. parasuis, which would be feasible and attractive for clinical samples diagnosis and epidemiological investigations.

为了研究副猪嗜血杆菌(Haemophilus parasuis)河南分离野毒株对昆明(KM)小鼠的致病性以及在感染KM小鼠体内组织器官的分布,将Hps血清5型标准菌株和河南分离野毒株采用灌胃和腹腔感染KM小鼠,观察2个Hps菌株对小鼠的致病性并测定Hps在感染小鼠体内组织器官的分布.结果显示,Hps河南分离野毒株和血清5型标准菌株腹腔、灌胃感染KM小鼠,在不同时间均出现发病,以腹腔感染发病症状严重、死亡率高,其中河南分离野毒株腹腔感染死亡率为2/7,血清5型标准菌株腹腔感染死亡率为1/7,死亡集中在攻毒后43～50 h;剖检死亡小鼠肉眼可见肺脏出血点、胸腔积液等病理变化.表明河南分离野毒株和标准菌株5型对KM小鼠均有致病性,为深入研究副猪嗜血杆菌的致病机制、免疫及防控该病提供科学依据.

高校教学中对应用技术的教育越来越重视,技能型人才在社会经济的发展中起到了越来越重要的作用,用人单位也对高校培养高素质、强能力、广适性人才的要求与呼声越来越高,这给高校人才培养提出了新的要求;适时代需要,高校教师不断探索、实践教育教学改革,在实践中启发和诱导大学生创新意识,培养其实践能力和创新能力是21世纪培养人才的必由之路,也是新形势下农业院校培养学生的重点内容.

A Staphylococcus aureus (S.aureus) was isolated from pigs suffered in pneumonia that can't be cured by antibiotic such as methicillin and vancomycin. It was demonstrated that baicalin, an active natural compound extracted from the traditional Chinese medicinal, possess antimicrobial activity. In the present study, we evaluate it efficacy in vitro and vivo against this isolated methicillin-resistant S.aureus (MRSA). Our findings demonstrated that baicalin can inhibit S. aureus growth in a dose-dependent manner and attenuate the biofilm formation. Scanning electron microscopies showed that cell membrane was damaged and accompany with contents leaks after treated with high concentration of baicalin. In addition, baicalin exerted inhibitory effects on the expression of S.aureus virulence-related factors. Moreover, baicalin treated mice had enhanced survival after a lethal dose of S.aureus infection compared with untreated mice. Simultaneously, the pathological tissue damage and bacterium burden were decrease in baicalin treated mice. These data demonstrated that baicalin displayed a high effectiveness in vitro and vivo against MRSA infection, suggesting that baicalin may potentially be used to treat MRSA infection.

何为人格魅力,《现代汉语词典》解释“魅力”为“很能吸引人的力量.”魅力是自身素质所引起的,散发出强烈的吸引力和感染力,这种力量会给周围的人很大的影响,会让人效仿、比较、赞叹,甚至迷恋,非权利影响力.《现代汉语辞典》解释“人格魅力”是个人的性格、气质、能力等特征的总和,个人的道德品质,人的能作为权利、义务主体的资格.高校教师人格魅力是指从事高等教育教学的道德品质,性格气质,渊博的知识,自身专业知识的厚集,这些对学生学习专业知识具有强悍的吸引力、号召力;教师教育的对象是新时代的大学生,是一个个充满鲜活力的生命,有着对生活的激情、对知识的渴望和对美好社会的向往,在教学活动中去引导、培养有责任有担当的接班人是每一位高校教师职责所在.

为了研究最佳的H1N1亚型流感病毒鸡胚增殖参数,本试验进行了孵化前种蛋的选择与保存、鸡胚孵化中各参数设定等因素对H1N1亚型流感病毒产毒量影响的研究.其中种蛋的选择与保存,主要考察了蛋重、蛋形指数、保存期、消毒时间及方法等因素,结果显示,蛋重为55～65 g,蛋形指数为1.30～1.35,种蛋保存期为1～4 d,保存温度为16～18℃,保存湿度为70％～80％,保存期种蛋的甲醛熏蒸消毒时间为30 min时,可以为H1N1亚型流感疫苗生产提供最佳的种蛋.孵化过程中孵化参数对H1N1亚型流感病毒产毒量的影响,本试验主要将孵化温度、湿度、翻蛋、通风等参数作为研究对象,结果显示在生产H1N1亚型流感疫苗时,最佳孵化参数设定为:温度1～7d为38.2℃、8～9 d为38.0℃、10 d为37.8℃,湿度1～10 d为65％～70％,翻蛋频率为1次/2 h,前后倾角各为45°,通风风门设定为1～5 d为4、6～10 d为5.本试验结果为H1N1亚型流感疫苗生产提供优质的鸡胚孵化技术,确保鸡胚尿囊液的质量和收获量.

由发病蛋鸡体内分离得到一株禽流感病毒,经PCR、血凝试验和免疫荧光鉴定为H9N2亚型,命名为A/Chicken/henanxy/2010(H9N2),病毒对SPF鸡无致病性,对其HA基因进行克隆测序,HA基因氨基酸裂解位点为PSRSSRGLF,符合低致病性禽流感的分类标准;并将HA基因和自GenBank读取的H9N2病毒的HA基因序列比较,表明河南分离株属于欧亚分支中的Y280-like分支,与A/Chicken/shandongHL/2010(H9N2)氨基酸同源性94.8％,可能是Y280-1ike个别核苷酸突变的一株.

Seven strains of bacteria were isolated from clinical cases of swine disease in lungs and trachea. The isolated bacteria was identified as Actinobacillus pleuropneumoniae (APP) serotypes 1 through bacterial cultivated, culture characteristic observation, morphology, biochemical, growth of satellite phenomenon, hemolysis and serotype identification. The antimicrobial susceptibility showed that the isolated A. pleuropneumoniae resistance to penicillins, semisynthetic antibiotics, p-lactam antibiotics, polypeptide antibiotic, polyene antibiotic, coumarin antibiotic, amphenicols antibiotics, beta-lactam/beta-lactam inhibitor compounds, cephalosporins, aminoglycosides, tetracyclines, fluoroquinolones, macrolides, sulfa drugs, sugar peptide drugs, furan drugs, ethlhydrocupreine; medium sensitive to cephalothin (CF), cefizime (CEX), ciprofloxacin (CIP), norfloxacin (NOR), ofloxacin (OFL); high sensitive to polymyxin B (PB), fosfomycin (FOS), ceftizoxime (CTZ), cefmetazole (CMZ), cefaclor (CEC), cefoperazone/sulbactam (CFP/SU), cefetamet (CTM), cefotaxime (CTX), ceftazidime (CAZ), cefodizine (CDZ), cefurosimc sodium (CXM), cefotaxime/clayulanic acid (CTX/CA), ceftazidime/clavulanic acid (CAZ/ CA), cefoxitin (FOX), cefepime (FEP), ceftriazone (CRO), aztreonam (AZT), minocycline. This result showed that A. pleuropneumoniae clinical strains had multi-drug resistance and resistance characteristics, which means that these isolated bacteria was multiple drug resistant strains.

采用超声裂解副猪嗜血杆菌分离株5型菌体上清作为包被抗原,初步建立副猪嗜血杆菌抗体的间接ELISA检测方法.通过棋盘滴定法筛选确定最佳反应条件:抗原包被浓度50 mg/L,37℃作用2h后4℃过夜包被,血清的稀释度为1∶320,抗原抗体反应时间为50 min,酶标二抗稀释度为1∶12 000,作用时间为30min,底物显色时间为15 min.经特异性、敏感性和符合性试验检测结果表明,建立的间接ELISA方法特异性和重复性良好,敏感性高,可用于副猪嗜血杆菌的检疫和流行病学调查.

Licorice is the most important herb used in the traditional Chinese (TCM) medicine. It was used to treat individuals with gastric or duodenal ulcers, bronchitis, cough, arthritis, adrenal insufficiency and allergies. The active substances and secondary metabolites from Licorice roots may be associated with a specific microbial agent. It is believed that many endophytic fungi from medically important plants can produce the same or similar secondary metabolites as the plant. Endophytic fungi are also thought to play an important role in plant communities by increasing fitness of the host plant. To clarify whether endophytic fungi are found in the licorice, fungi were isolated from Licorice root and characterized morphologically and using molecular identification. The morphological examination showed that the endophytic fungi from Licorice were Aspergillus and Chaetomium species. Further identification was achieved by sequence similarity comparison and phylogenetic analysis of the ITS regions. Results showed that strains L10L2 and L4 belong. to Aspergillus species, L3 identified to Chaetomium. A neighbor-joining tree showed the relationships between the isolate's sequence data and the closest identified relatives from GenBank. These fungi could have significance as a source of pharmaceutical natural products.

The first-line teaching experience is summarized for many years,which changes the traditional teaching style.The new teaching style is embedded in a variety of teaching methods.Such as Problem Navigation Act, multimedia seminar, case study teaching etc.The study results showed the methods can mobilize students'initiative,so that students become the subject in the study,which enhances students'abilities in discovering problems and solving them.At the same time,it effectively improved the teachers'comprehensive ability and teaching level,and promoted the teachers'comprehensive quality,to achieve the purpose of learning while teaching.

副猪嗜血杆菌病是由副猪嗜血杆菌（ Hps）引起的一种细菌性传染病。从疑似发生副猪嗜血杆菌病的河南新乡某猪场采集病猪呼吸道分泌物和心包积液，并对样品进行细菌分离培养、形态观察、培养特性、生化试验鉴定和动物试验。结果表明，该试验成功分离到了副猪嗜血杆菌，药敏试验显示，分离菌株对庆大霉素、环丙沙星、氧氟沙星、诺氟沙星和头孢哌酮高敏，对复方新诺明中敏，对青霉素、链霉素、林可霉素耐药；对金银花及其组成的复方敏感，且复方作用效果优于单方。

为了研究H9N2流感病毒在鸡胚中的繁殖规律及其血凝活性,试验用H9N2河南分离毒株接种鸡胚,对收获的尿囊液进行10倍系列稀释,选取不同浓度尿囊液分别接种不同日龄鸡胚,测定不同日龄鸡胚接种病毒后尿囊液产量及其血凝效价,探讨影响毒株血凝活性的因素.结果表明:10日龄鸡胚接种1×10-5病毒时收获的尿囊液产量最多且血凝效价最高;筛选分析影响H9N2病毒血凝活性因素时发现,缓冲液的pH值为3.0 ～5.5时发生溶血,水浴温度为70 ～ 80℃时几乎完全丧失活性.

在293细胞中瞬时表达A/Chicken/Henan/1001/2010(H9N2) NS1基因蛋白,并对表达蛋白活性进行测定.试验利用PCR技术从NS1-T载体质粒上扩增NS1基因,将其克隆至pCAGGS载体,构建重组质粒NS1-pCAGGS;经酶切和测序鉴定正确后,重组质粒NS1-pCAGGS与脂质体按照一定比例混合后转染293细胞,用间接免疫荧光方法对瞬时表达细胞进行荧光信号反应.结果显示,禽流感NS1基因可以很好地在293细胞中瞬时表达,具有良好的反应原性.

In order to study therapeutic efficacy that different drugs on pathogenic Escherichia coli, the experiment selected 40 pigs 7 days old from a pig farm in natural infection in Jiyuan city of Henan and were randomly divided into 4 groups, each group of 10 head. Through to the diagnosis of pathogenic Escherichia coli were isolated and cultured, biochemical identification, drug sensitive test, selected from three kinds of drugs in the same environment. Same feeding and management conditions, respectively, for the three group piglets were treated, the management conditions, respectively, for the three group little pigs were treated. The experimental results show that, the cure rate of ceftiofur is 90%, the cure rate of polymyxin is 60%, the cure rate of enrofloxacin is 50%.