Isolation,Identification and Analysis of Serotype and Antibiotic Susceptibility of Pathogenic Salmonella from Chickens

To investigate the situations of predominant strain and antibiotic resistance of pathogenic Salmonella from chickens in Guangxi Zhuang Autonomous region, Salmonella was pre-enriched and isolated from tissues of clinical suspected chickens, and the Salmonella isolates were identified by biochemical test using ID 32E System for the identification of Enterobacteriaceae of VITEK System ATB Expression, serotypes were determined by slid agglutination test, antibiotic susceptibility was analyzed by ATB VET Susceptibility Test Strip of Enterobacteriaceae antibiotics.The results showed that 34 Salmonella strains were obtained from 310 clinical samples, of which 1 strain belonged to A serogroup, 1 to C2 serogroup, 15 to B serogroup, 14 to D serogroup and 3 to untyped serogroup, and S.typhimurium of B serogroup and S.pullorum of D serogroup were the predominant serotypes.All Salmonella isolates were sensitive to meropenem, imipenem, spectinomycine and apramycine, and the resistance rates to chloramphenicol, kanamycine and gentamicine were less than 10%, and the resistance rates to amoxicillin, streptomycine, flumequine, oxolinique, sulfamethizol, tetracycline and nitrofurantoine ranged between 50% and 90%, while the resistance rates to penicilline, oxacilline, fusidique, rifampicine and metronidazole reached to 100%.The results indicated that the serotype distribution of pathogenic Salmonella from chickens exhibited regional characteristics, and S.typhimurium and S.pullorum were the predominant serotypes in Guangxi Zhuang Autonomous region, and antibiotic resistance of Salmonella isolates was very serious which should be highly concerned.     

扭角羚肺炎克雷伯氏菌的分离鉴定

本试验旨在报告四川省野生扭角羚自然生存过程中致病性细菌的检测及生物学特征。试验对发病死亡扭角羚病变组织进行细菌学检查及分离,对分别从肝脏、脾脏和肺脏分离得到的4株优势菌进行形态特征、生化特性和16SrDNA分子鉴定,判定为肺炎克雷伯氏菌。这4株菌对供试小白鼠均有致病性。测其LD50,分别为2.9×107、2.9×107、4.5×107和1.1×108 CFU/只。4株分离菌对供试的先锋霉素等敏感,对阿米卡星等中度敏感,对红霉素等耐药。推测这4株肺炎克雷伯氏菌是导致扭角羚死亡的主要病原菌。     

Isolation and Identification of one Strain of H9 Subtype Avian Influenza Virus and Study on its Biological Characteristics

In 2013,one case of suspected H9 subtype avian influenza occurred in a chicken farm of Jilin povince.Clinical samples were collected from the diseased farm,inoculated into the allantoic cavity of 9-day-old SPF chicken embryo,and then one strain of virus was isolated.The results of HA test,HI test and molecular biology test all showed that the isolate belonged to H9 subtype avian influenza virus (AIV).The HA cleavage site of the isolate was RSSR↓GLF,which was consisted with the molecular characteristic of low pathogenic AIV.The HA peptide chain had 9 potential glycosylation sites which were same as other isolates of recent years.The isolate had 8 receptor binding sites,including 234 receptor binding site by glutamine (Q) mutating into threonine (T).The phylogenetic tree revealed the isolate belonged to Eurasian lineages and it had far genetic relationship with the earliest domestic isolate (A/Chicken/Beijing/1/94(H9N2)),but had close genetic relationship with the representative strain (A/Chicken/Guangxi/55/2005(H9N2)) of major epidemic branch since 2007.We prepared an inactivated oil-emulsion vaccine of the isolate,and then vaccinated SPF chicken.21 days after vaccination,the HI titer of chicken serum antibody reached up to 10log2.The result suggested the isolate had good immunogenicity.     

江西永修凤凰山桃实腐病病原鉴定

桃实腐病是江西永修凤凰山桃园发生的一种新的重要病害,其主要危害挂果期的果实,严重影响桃产量和品质。病果主要表现为脐部变褐腐烂,并在病部表面密生黑色小点（分生孢子器）,病果易脱落。本文对病果进行了组织分离,对获得的病菌纯培养进行了回接,同时对病菌的形态及大小观察和测量,结果表明引起凤凰山桃实腐病的病原菌为核果果腐拟茎点菌（Phomopsis amygdalina Canonaco）。     

雏番鸭病毒性肝炎病毒的分离与鉴定

从洛江区临床发病鸭分离到2株病毒,2株分离病毒能够致死鸡胚,能被Ⅰ型鸭病毒性肝炎标准强毒高免血清特异性中和。分离毒株在鸡胚上传代培养,并测定ELD50分别为10-6.32/0.2 m L、10-5.49/0.2 m L;经动物回归试验,对1日龄雏番鸭的致死率分别为80%和70%,雏番鸭出现明显的角弓反张姿态,剖检可见肝脏出血斑、出血点,为鸭病毒性肝炎的典型症状,表明分离到的病毒为Ⅰ型鸭病毒性肝炎病毒(DHV-I)。     

貉源犬瘟热病毒CDV-ZC株分离鉴定、全基因测序与分析

应用非洲绿猴肾细胞(Vero)从山东省诸城市疑似犬瘟热(canine distemper,CD)感染貉的肝脏、脾脏、肺脏等组织病料的研磨上清液中分离出1株病毒。分离株经Vero细胞传至第4代出现典型的细胞病变效应(CPE),经毒力测定、血清学、RT-PCR检测及测序、回归动物试验证明为犬瘟热病毒(Canine distemper virus,CDV),命名为CDV诸城分离株(CDV-ZC)。采用RT-PCR方法分段克隆分离株的全基因序列并测序,测序成功的各序列依次拼接得到全基因序列并进行序列比对。结果表明,CDV-ZC株全基因(KJ994343)与标准美洲型犬瘟热强毒株A75/17核苷酸同源性高达96.5%,而与疫苗株Onderstepoort、CDV3等亲缘关系较远,同源性为91.6%~92.0%。血凝素(H)基因序列分析表明,CDV-ZC株与国内野毒株LN(13)2、GP株等以及日本野毒株UENO、HAMA等共同归属于Asia-1型,在H蛋白信号淋巴细胞激活因子(SLAM)受体结合区即542~544位增加了1个潜在N-连接糖基化位点(N-X-S/T)。致病性强毒株CDV-ZC的成功分离及全基因序列测定加深了我们对当前中国CDV流行株遗传变异情况的了解,为CD的有效预防、诊断及控制提供理论依据。     

羊流产衣原体的分离鉴定及间接免疫荧光法检测

对收集的疑似衣原体感染的羊流产胎儿样本接种鸡胚卵黄囊进行分离培养,通过PCR-RFLP(限制性片段长度多态性分析)方法鉴定为流产衣原体。将收集到的流产衣原体阳性样本接种Hela229细胞,应用基于流产衣原体MOMP单克隆抗体的间接免疫荧光法,在Hela229细胞浆内可见呈绿色荧光的衣原体包涵体,进一步在病原学水平上确定发生在甘肃省肃南县羊流产的病原为流产衣原体。     

7株牛源A型巴氏杆菌的分离鉴定及耐药性分析

对2014年沈阳地区患有呼吸系统疾病的育肥牛场进行病原分离,得到7株分离菌,对其进行培养形态及染色观察、病理组织切片观察、动物致病性观察及16S序列分析,初步鉴定为牛多杀性巴氏杆菌(Pm)。利用Pm种特异性引物及荚膜血清特异性引物进行PCR扩增,均得到目的基因条带。对分离菌进行药敏试验的结果显示,5株分离菌株已对阿米卡星、新诺明产生较强耐药性;7株分离菌均对氟苯尼考、恩诺沙星、四环素及氨苄西林敏感。由此确定,分离菌株均为牛荚膜A型巴氏杆菌,且已具有一定耐药性。     

紫花苜蓿根腐病原菌--拟枝孢镰刀菌的鉴定及其生物学特性研究

本文对从内蒙古赤峰巴雅尔草业基地紫花苜蓿根部分离到的菌株 BYE27-2-5进行了致病性测定、形态学和rDNA-ITS 序列分析鉴定及生物学特性测定。结果表明,该菌菌丝生长、产孢和孢子萌发的最适温度分别为25,30和28℃;pH 值5．0～11．0均宜于菌丝生长,产孢最适 pH 8．0,孢子萌发最适 pH 7．0;光暗交替利于菌丝生长,光照利于产孢和孢子的萌发;孢子在相对湿度低于75％不萌发,高于95％萌发较快;葡萄糖蛋白胨培养基利于菌丝生长和孢子萌发,马铃薯蔗糖琼脂培养基利于产孢;碳源中葡萄糖利于菌丝生长和孢子萌发,蔗糖利于产孢;氮源中蛋白胨利于菌丝生长,酵母膏利于产孢和孢子萌发;菌丝致死温度为54℃（10 min）,分生孢子的致死温度为48℃（10 min）;根据其形态特征和 rDNA-ITS 序列分析结果,鉴定其为拟枝孢镰刀菌（Fusarium sporotrichioide ）。     

野生老芒麦种质资源鉴定与基部叶鞘绒毛变异分析

为更好地利用老芒麦(Elymus sibiricus L.)野生资源,本研究从我国青藏高原、西北、华北、东北地区以及国外部分地区共采集了1 723份野生披碱草属种质资源,通过表型特征观测和流式细胞仪检测,共鉴定出了990份野生老芒麦种质,246份垂穗披碱草种质,并测得老芒麦的DNA含量在5.86～7.30 Gb之间,平均DNA含量为6.66 Gb;垂穗披碱草材料的DNA含量在9.50～10.36 Gb之间,平均DNA含量为9.97 Gb。研究结果显示,在西北、华北以及东北地区的部分野生老芒麦材料在苗期基部叶鞘出现了绒毛,分析发现该表型出现的概率与经度和纬度呈极显著正相关关系(P<0.01),与海拔、年年均气温以及年平均降雨量呈极显著负相关关系(P<0.01)。表明老芒麦苗期基部叶鞘绒毛的有无与环境因子之间存在着必然的联系,可能是老芒麦种质应对不同环境类型的适应性机制之一。本研究为老芒麦的形态鉴定提供了新的依据,为老芒麦种质资源挖掘和育种应用提供了材料基础。