一株鸡新城疫病毒的分离鉴定及其油乳剂灭活苗免疫效果评价

从河南某种鸡场发病鸡群中分离出1株毒株,经HA、HI和动物回归试验,确定分离的病毒为NDV,命名为HNYD株。经过测定,HNYD株鸡胚半数致死量（ELD50）为10^90/0．1mL;鸡胚最小致死量的平均致死时间（MDT）为53．8h;对1日龄SPF雏鸡脑内致病指数（ICPI）为1．86,提示HNYD株为强毒力株。血清交叉反应试验显示HNYD株和La Sota株存在一定的抗原性差异。用HNYD株油乳剂灭活苗对HNYD株和La Sota株的保护率分别为90％和100％,明显优于La Sota株油乳剂灭活苗,提示HNYD株制备的疫苗对流行毒株和经典毒株的攻击均能很好的保护。     

Rapid detection of <Emphasis Type="Italic">Peste des Petits Ruminants</Emphasis> (PPR) virus antigen in Sudan by agar gel precipitation (AGPT) and haemagglutination (HA) Tests

AGPT and HA tests were employed for rapid diagnosis of PPRV infection in sheep and goats in Sudan. Forty lymph nodes and spleen samples from suspected cases of PPR in both sheep and goats were examined by AGPT and HA tests for detection of PPRV antigen. Viral antigen was detected from (77.5%) of the samples tested by AGPT and (92.5%) tested by HA test. The results of both tests revealed that HA test was more sensitive than AGPT for detection of PPRV antigen (Kappa statistics 0.4366). Another advantage of the HA test over AGPT was that it can differentiate PPRV from RPV. Thus the HA test represents a quick, easy, simple, cheap and reliable confirmatory test for the diagnosis of PPR and differential diagnosis of PPRV and RPV. The HA test was carried out using chicken, goat and pig RBCs. Chicken RBCs were found to be the most sensitive for detection of PPRV antigen, followed by goat then pig RBCs. The HA time when using chicken RBCs was 20–25 minutes, using goat RBCs was 25–30 minutes and using pig RBCs was 40–45 minutes. The distribution of PPR infection in four different regions of Sudan was investigated.     

毛皮动物源大肠杆菌毒力和耐药基因检测及药物敏感性分析

为了解秦皇岛昌黎地区毛皮动物大肠杆菌毒力基因和耐药基因的分布及药物敏感性情况,以60只临床患病毛皮动物为试验菌株的分离宿主,采用常规分离纯化、生化鉴定以及分子生物学的方法对病原菌进行鉴定,动物致病性试验检测其致病性,PCR方法检测毒力基因和耐药基因,K-B法对常用西药进行药物敏感性检测,琼脂平板打孔法对中草药药物敏感性进行检测,试管二倍稀释法检测中草药的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。结果显示:此次分离到的28株大肠杆菌均使小鼠发病死亡;其中,24株大肠杆菌检测到irp2、fyuA毒力基因,检出率为85.7%;5株大肠杆菌临床分离菌株检出Ler、eaeA毒力基因,检出率为17.9%;耐药基因blaTEM、aadA1、blaCTX-M、blaTEM1、blaSHV-1、strA-strB、blaPSE1和blaOXA1的检出率分别为85.7%,89.3%,75.0%,71.4%,25.0%,24.1%,14.3%,14.3%;西药药敏试验结果显示,28株菌对链霉素、头孢拉定、多西环素、复方新诺明、林可霉素、替米考星、青霉素同时耐药;中草药药敏试验结果显示,诃子、乌梅、五味子对这28株大肠杆菌均具有较好的体外抑菌效果,抑菌圈直径为18～31 mm,MIC和MBC为15.63～500.00 g/L。     

1株山羊源海藻糖比伯斯坦杆菌的分离鉴定及药敏试验

为确定引起山羊肺炎的病原,本研究从病死山羊肺脏中分离纯化获得1株细菌,通过生化试验和分子生物学方法鉴定为海藻糖比伯斯坦杆菌。利用生物信息学软件MEGA6.0对巴氏杆菌科5个属细菌的白细胞毒素基因(lktA)和RNA聚合酶β亚基基因(rpoB)分别构建遗传系统发育树,可知分离株与海藻糖比伯斯坦杆菌(AF314526.2、CP003745.1、CP006955.1等)的lktA基因相似性最高,位于同一簇内;分离株和海藻糖比伯斯坦杆菌(CP006956.1、JACI01000002.1、CP003745.1等)的rpoB基因聚类为一簇,相似性最高;而与和葡萄糖苷曼氏杆菌(KU986490.1)的rpoB基因相似性最低。药敏试验显示,该菌对头孢噻肟、头孢他啶和环丙沙星等抗菌药物最敏感,对多西环素、阿莫西林和林可霉素等耐药。本研究为该菌致病机理的研究与疫苗的研发奠定了基础,也为临床用药提供了参考依据。     

狐和貉源大肠埃希菌药敏试验及耐药基因检测

为了解秦皇岛昌黎县狐和貉源致病性大肠埃希菌耐药性及耐药基因的情况,以昌黎县分离的15株狐和貉源致病性大肠埃希菌为试验菌株,采用药敏纸片法对抗菌药物进行药敏试验,并对四环素类耐药基因tetA、tetB、tetC、tetD,氨基糖苷类耐药基因strA-strB、aadA1,磺胺类耐药基因sul1、sul2、sul3进行PCR检测。结果显示,15株大肠埃希菌分离菌株对土霉素、多西环素、头孢拉定、新霉素、青霉素、磺胺间甲氧嘧啶、磺胺二甲氧嘧啶的耐药率均为100%;对阿莫西林、庆大霉素、头孢曲松、阿米卡星、大观霉素、链霉素、复方新诺明、氨苄西林、环丙沙星耐药率分别为93.33%、53.33%、33.33%、73.33%、86.66%、86.66%、86.66%、66.67%和66.67%。9种耐药基因tetA、tetB、tetC、tetD、strA-strB、aadA1、sul1、sul2、sul3的检出率分别为0、46.67%、26.67%、13.33%、40%、100%、53.33%、46.67%和53.33%,其中四环素类耐药基因以tetB的检出率最高,氨基糖苷类耐药基因以aadA1的检出率最高,磺胺类耐药基因的检出率相差不大。说明15株狐和貉源大肠埃希菌耐药谱广,耐药性高,耐药基因流行普遍。     

Length of lags in responses of milk yield and somatic cell score on test day to heat stress in Holsteins

We used daily records from provincial Japanese weather stations and monthly test‐day records of milk production to investigate the length of the lags in the responses of cows’ milk yield and somatic cell score (SCS) to heat stress (HS). We also investigated the HS thresholds in milk yield and SCS. Data were a total of 17,245,709 test‐day records for milk and SCS in Holstein cows that had calved for the first time between 2000 and 2015, along with weather records from 60 weather stations. Temperature–humidity index (THI) values were estimated by using average daily temperature and average daily relative humidity. Adjusted THI values were calculated by using temperature, relative humidity, wind speed, and solar radiation. The model contained herd, calving year, month of test day, age group, days in milk, and THI as a fixed effect. THIs for each day from 14 days before the test day until the test day were used to represent the HS effects. The HS occurring 3 days, and between 8 and 10 days, before the test day had the greatest effect on the milk yield and SCS, respectively. The threshold THI values for the HS effect were about 60–65 for both traits.     

小反刍兽疫病毒N蛋白抗体与H蛋白抗体在羊体内代谢消长规律的研究

为研究小反刍兽疫病毒N蛋白抗体与H蛋白抗体在羊体内的代谢消长规律,试验通过建立小反刍兽疫N蛋白双抗原夹心ELISA抗体检测方法、H蛋白阻断ELISA抗体检测方法与血清中和抗体试验方法,分别检测了羊免疫疫苗后体内N蛋白抗体与H蛋白抗体在每个免疫阶段的代谢消长变化。结果表明:羊免疫小反刍兽疫疫苗后,血清内的N蛋白抗体与H蛋白抗体在6~8周时血清抗体效价较高,对应的羊体内中和抗体效价为1∶512,且效价至少可以持续到10周以上,2种抗体具有一定消长代谢规律的相关性。该研究结果也为以N抗原与H抗原为基础建立相应的抗原捕获ELISA方法、竞争ELISA检测方法、间接ELISA检测方法以及双抗原夹心ELISA检测方法奠定了一定的理论依据。     

穿心莲超声提取工艺研究

优选穿心莲药材的提取工艺。通过单因素和正交试验,测定穿心莲内酯、脱水穿心莲内酯、新穿心莲内酯三种二萜内酯类的总含量,确定最佳超声提取工艺。确定穿心莲药材的最佳超声提取工艺为乙醇浓度60%,超声时间1h,料液比为1∶15,此条件下穿心莲二萜内酯类的总含量为2.27%。该试验确定的超声提取工艺稳定可行,方法可靠,可为穿心莲有效成分的提取提供理论依据。     

一例鸡源致病型大肠杆菌的分离鉴定

为进行一例鸡源致病型大肠杆菌的分离鉴定,无菌采集疑似大肠杆菌患病蛋鸡肝脏、脾脏等组织,进行病原菌的分离培养。利用培养特性检查、菌落形态观察、革兰氏染色鉴定、生化特性考察、PCR鉴定、测序分析以及动物致病性试验对分离菌株进行鉴定,并同时进行药敏试验。结果显示,分离菌株为一株致病型大肠杆菌,对青霉素、氟苯尼考、阿莫西林等多种抗生素产生耐药性,对丁胺卡那敏感。建议使用丁胺卡那霉素对大肠杆菌进行防治。