猪胸膜肺炎放线杆菌的分离与鉴定

吉林省某规模化养猪场送检疑似猪传染性胸膜肺炎的死猪病料,采用不同培养基进行病原菌分离,结果分离出3株疑似胸膜肺炎放线杆菌。通过对分离菌进行镜检、培养特性试验、生化特性试验、PCR鉴定,确定为3株均为胸膜肺炎放线杆菌,致病性试验结果表明,分离的胸膜肺炎放线杆菌具有强致病性。     

洛阳某猪场猪传染性胸膜肺炎放线杆菌的分离鉴定

为确定洛阳某猪场疑似猪传染性胸膜肺炎的病原种类及其血清型,为其临床科学防疫和合理用药提供参考方案,本试验无菌采集了病猪肺脏等病料,采用病菌培养后形态观察、革兰氏染色、生理生化特性试验、PCR诊断等技术对病原菌进行实验室分离鉴定,通过基因测序和同源性分析方法对其进行确诊,通过动物攻毒试验和药敏试验分析分离菌株的致病性和耐药性,采用5对分型引物对分离菌进行基因分型以确定病原菌血清型。结果显示,分离菌2007菌落形态为光滑、透明、边缘整齐、圆形、隆起的小菌落,革兰氏染色特征为阴性杆菌且呈多型性,具有猪传染性胸膜肺炎放线杆菌（APP）的典型培养特征,且生化结果符合猪传染性胸膜肺炎放线杆菌的生化特性。同源性分析结果显示,分离菌株2007与猪传染性胸膜肺炎放线杆菌菌株FJ848573.1同源性为99.4%,说明分离株2007是猪传染性胸膜肺炎放线杆菌。动物攻毒试验结果显示,分离菌株2007对小鼠有强致病性。药敏试验结果显示,分离株2007对泰妙菌素、氨苄西林和头孢噻呋钠等高敏,对替米考星、红霉素、氟苯尼考等20种抗菌药完全耐药,对34种抗生素的耐药率高达76.5%,具有较强的多重耐药性。分离菌2007基因分型扩增到目的基因片段与血清型5型结果相符。本试验结果为发病猪场提出了解决该病的具体防治措施,并最终获得了良好的效果,为猪传染性胸膜肺炎放线杆菌临床耐药折点的制定提供了一些参考。     

洛阳某猪场猪传染性胸膜肺炎放线杆菌的分离鉴定

为确定洛阳某猪场疑似猪传染性胸膜肺炎的病原种类及其血清型,为其临床科学防疫和合理用药提供参考方案,本试验无菌采集了病猪肺脏等病料,采用病菌培养后形态观察、革兰氏染色、生理生化特性试验、PCR诊断等技术对病原菌进行实验室分离鉴定,通过基因测序和同源性分析方法对其进行确诊,通过动物攻毒试验和药敏试验分析分离菌株的致病性和耐药性,采用5对分型引物对分离菌进行基因分型以确定病原菌血清型。结果显示,分离菌2007菌落形态为光滑、透明、边缘整齐、圆形、隆起的小菌落,革兰氏染色特征为阴性杆菌且呈多型性,具有猪传染性胸膜肺炎放线杆菌(APP)的典型培养特征,且生化结果符合猪传染性胸膜肺炎放线杆菌的生化特性。同源性分析结果显示,分离菌株2007与猪传染性胸膜肺炎放线杆菌菌株FJ848573.1同源性为99.4%,说明分离株2007是猪传染性胸膜肺炎放线杆菌。动物攻毒试验结果显示,分离菌株2007对小鼠有强致病性。药敏试验结果显示,分离株2007对泰妙菌素、氨苄西林和头孢噻呋钠等高敏,对替米考星、红霉素、氟苯尼考等20种抗菌药完全耐药,对34种抗生素的耐药率高达76.5%,具有较强的多重耐药性。分离菌2007基因分型扩增到目的基因片段与血清型5型结果相符。本试验结果为发病猪场提出了解决该病的具体防治措施,并最终获得了良好的效果,为猪传染性胸膜肺炎放线杆菌临床耐药折点的制定提供了一些参考。     

血清6型猪传染性胸膜肺炎放线杆菌的分离与鉴定

从广东某猪场出现呼吸困难、肺脏有出血性纤维渗出性病变的病死猪中分离出1株有多形性趋向的革兰阴性球杆菌,卫星现象观察、CAMP试验及V因子依赖性试验均为阳性,用PCR方法能扩增出各血清型猪传染性胸膜肺炎放线杆菌均有的apxⅣA基因长约422 bp的特异片段,小白鼠攻毒试验显示其有致病性,血清学检测表明该分离菌为血清6型猪传染性胸膜肺炎放线杆菌。     

猪传染性胸膜肺炎的病例分析

猪传染性胸膜肺炎是由胸膜肺炎放线杆菌引起猪的一种高度传染性呼吸道疾病,又称为猪接触性传染性胸膜肺炎。分离自山东某养殖场的疑似病例,并对典型的疑似猪传染性胸膜肺炎病例进行放线杆菌的分离鉴定。分离到5株细菌为多形态、革兰氏阴性杆菌;生化试验鉴定结果表明所分离的上述5株菌株为猪传染性胸膜肺炎放线杆菌。文内介绍了猪传染性胸膜肺炎的发病情况以及药敏实验结果,为生产优质猪肉提供有效的理论支持。     

猪传染性胸膜肺炎放线杆菌血清2型PCR程序优化及分子鉴定

猪接触性传染性胸膜肺炎（porcine contagious pleuropneumonia,PCP）又称坏死性胸膜肺炎,是由胸膜肺炎放线杆菌引起的一种高度接触传染性、致死性呼吸道传染病。以急性出血性纤维素性肺炎和慢性纤维素性坏死性胸膜炎为主要特征。各种年龄、性别的猪对本病均易感,急性者死亡率高,慢性者常能耐过。猪传染性胸膜肺炎放线杆菌有15个血清型,不同血清型之间没有或仅有较弱的交叉保护作用,这给猪传染性胸膜肺炎放线杆菌的诊断防治和免疫预防带来了很大的困难。因此建立一种快速血清型分子鉴定方法尤为重要。为了快速、简便的建立猪传染性胸膜肺炎放线杆菌血清型分子鉴定的方法,本研究所从临床发病疑似病例猪肺脏和气管中分离到的放线杆菌,首先经过血清型鉴定,确定部分血清型为2型,为了确定血清型鉴定的准确性,在此基础上,采用分子鉴定的方法,对这些菌株进行鉴定,确定分离到的11株菌为猪胸膜肺炎放线杆菌血清2型。这为猪传染性胸膜肺炎放线杆菌血清型分子鉴定及防治提供了理论依据,为今后临床血清型定型提供了一种简便方法。     

血清6型猪传染性胸膜肺炎放线扦菌的分离与鉴定

从广东某猪场出现呼吸困难、肺脏有出血性纤维渗出性病变的病死猪中分离出1株有多形性趋向的革兰阴性球杆菌，卫星现象观察、CAMP试验及V因子依赖性试验均为阳性，用PCR方法能扩增出各血清型猪传染性胸膜肺炎放线杆菌均有的apxIVA基因长约422bp的特异片段，小白鼠攻毒试验显示其有致病性，血清学检测表明该分离茵为血清6型猪传染性胸膜肺炎放线杆菌。     

一株猪传染性胸膜肺炎放线杆菌的分离与药敏试验

从新乡地区某猪场疑似猪传染性胸膜肺炎病猪体内分离到一株致病菌。通过对分离菌株的形态学检查、生化实验、卫星生长现象、溶血试验及动物实验,证明该分离菌为胸膜肺炎放线杆菌,药敏试验结果表明,所分离菌株对氟苯尼考和头孢噻呋最敏感,可作为新乡地区猪传染性胸膜肺炎病的首选药物。     

猪胸膜肺炎放线杆菌PCR诊断方法的建立与应用

根据胸膜肺炎放线杆菌ApxⅣ基因设计1对引物，扩增特异的650bp棱酸片段，建立了应用PCR检测猪胸膜肺炎放线杆菌的方法。特异性试验结果表明，12个血清型的放线杆菌参考菌株均能扩增出650bp特异性的核酸片段，而大肠杆菌、多杀性巴氏杆菌、猪肺炎支原体、伤寒沙门氏菌和支气管败血性波氏杆菌的扩增结果均为阴性。敏感性试验结果表明，PCR的最低检出限量为500个放线杆菌。利用建立的PCR检测方法对22株从山东省不同地区分离的疑似胸膜肺炎放线杆菌菌株进行检测．结果14株为阳性。对感染猪病变组织的检测结果表明，病变部位不同，胸膜肺炎放线杆菌的检出率不同，其中以扁桃体的检出率最高。     

猪胸膜肺炎放线杆菌和副猪嗜血杆菌的鉴别诊断及血清型鉴定

利用PCR技术、琼脂扩散试验、糖发酵试验和间接血凝试验(IHA)对野外分离到的可疑猪胸膜肺炎放线杆菌和副猪嗜血杆菌进行了诊断和血清型鉴定.PCR鉴定分离物HS1580为副猪嗜血杆菌,分离物HS1582为猪胸膜肺炎放线杆菌;生化试验表明分离物HS1581和HS1582为猪胸膜肺炎放线杆菌;血清型鉴定分离物HS1580不属于被检的14个血清型之列,HS1581为App血清15型,HS1582为App血清7型.试验结果表明,综合使用PCR等技术可快速、准确地对这两种传染性细菌进行鉴别诊断和血清型鉴定.     

电感耦合等离子-质谱法测定饲料中钠、镁、铬、锰、铁、铜、锌、砷、硒、镉和铅

应用电感耦合等离子-质谱技术(ICP-MS),建立饲料中钠、镁、铬、锰、铁、铜、锌、砷、硒、镉和铅等元素的测定方法。对饲料样品的前处理方法、仪器工作参数和11种元素标准曲线进行优化;并以加标回收、分析方法比对和重复测试说明方法的准确性和精密性。方法在0～1 000 ng/ml范围内线性良好,仪器检出限为0.557 7～5.072 ng/ml,具有良好的精密度,其回收率在88.1%～104.4%之间,相对标准偏差小于5.0%。同时与原子吸收和原子荧光方法进行比对,测定结果相近。所建立的方法简单、快速,可替代原子吸收和原子荧光方法测定饲料中的11种金属元素,为饲料的质量控制提供理想的元素分析方法。     

Supplementation of broodmares with copper, zinc, iron, manganese, cobalt, iodine, and selenium

In experiment 1, 6 pregnant mares received a concentrate that contained a trace mineral premix that provided 14.3 mg Cu, 40 mg Zn, 28 mg Fe, 28 mg Mn, 0.08 mg Co, 0.16 mg I, and 0.16 mg Se/kg concentrate (group A). Seven mares received the same concentrate plus 502 mg Zn and 127 mg Cu once daily (group B). No differences (P > .05) in foal growth data, or Cu, Zn, and Fe concentrations of mare milk, mare serum, or foal serum were observed. In experiment 2, 6 pregnant mares received the same concentrate as group A (group C), and 8 mares received the same concentrate fortified with 4× the trace mineral premix (group D). Group C mares had higher serum Zn concentration at 1 day (P < 0.01) and 56 days (P < 0.04). Group C mares had higher milk Fe concentration at 28 days (P < .01), and group D mares had higher milk Cu concentration at 56 days (P < .01). Group C foals had higher serum Cu concentration at 14 days (P < .03). The results from this study provide no evidence to indicate that supplementing late gestating and lactating mares with higher dietary trace mineral levels than those recommended currently by NRC has any influence on foal growth and development, or on the Cu, Zn, and Fe concentrations of the mare milk, mare serum, or foal serum.     

Comparisons of Angus, Charolais, Galloway, Hereford, Longhorn, Nellore, Piedmontese, Salers, and Shorthorn breeds for weight, weight adjusted for condition score, height, and condition score of cows

Breed differences for weight (CW), height (CH), and condition score (CS) were estimated from records (n = 12,188) of 2- to 6-yr-old cows (n = 744) from Cycle IV of the U.S. Meat Animal Research Center's Germplasm Evaluation (GPE) Program. Cows were produced from mating Angus and Hereford dams to Angus, Hereford, Charolais, Shorthorn, Galloway, Longhorn, Nellore, Piedmontese, and Salers sires. Samples of Angus and Hereford sires were 1) reference sires born from 1962 through 1970 and 2) 1980s sires born in 1980 through 1987. The mixed model included cow age, season of measurement and their interactions, year of birth, pregnancy-lactation code (PL), and breedgroup as fixed effects for CW and CS. Analyses of weight adjusted for condition score included CS as a linear covariate. The model for CH excluded PL. Random effects were additive genetic and permanent environmental effects associated with the cow. Differences among breed groups were significant (P < 0.05) for all traits and were maintained through maturity with few interchanges in ranking. The order of F1 cows for weight was as follows: Charolais (506 to 635 kg for different ages), Shorthorn and Salers, reciprocal Hereford-Angus (HA) with 1980s sires, Nellore, HA with reference sires, Galloway, Piedmontese, and Longhorn (412 to 525 kg for different ages). Order for height was as follows: Nellore (136 to 140 cm), Charolais, Shorthorn, Salers, HA with 1980s sires, Piedmontese, Longhorn, Galloway and HA with reference sires (126 to 128 cm). Hereford and Angus cows with reference sires were generally lighter than those with 1980s sires. In general, breed differences for height followed those for weight except that F1 Nellore cows were tallest, which may in part be due to Bos taurus-Bos indicus heterosis for size.     

液相色谱-串联质谱法测定动物尿液中11种β-受体激动剂残留量的不确定度评估

采用液相色谱-串联质谱法测定动物尿液中11种β-受体激动剂残留量,对标准溶液、体积、质谱峰面积、浓缩过程及回收率等测定不确定度因素进行了分析,通过评定各不确定度分量及标准不确定度,得出11种β-受体激动剂的扩展不确定度在0.7 ～ 1.1 ng/mL范围内.由各因素对合成不确定度的贡献比分析可知,影响较大的因素为试验回收率及标准溶液浓度.     

Granulocyte isolation from whole blood of goat, sheep, cattle, horse, dog, pig, and man

A simple two step procedure for the isolation of caprine, ovine, bovine, equine, canine, porcine and human peripheral blood granulocytes is described. After enrichment of granulocytes by centrifugation, contaminating erythrocytes are lysed hypotonically. Recovery, purity, and viability of the granulocyte suspensions are determined. FACScan analysis of the cell suspensions measuring cellular size by forward and sideward light scatter is compared with the corresponding analysis of whole blood leukocytes. Constituencies of the isolated cell suspensions and loss of granulocyte subpopulations through isolation procedure is discussed with regard to granulocyte function assays.