细胞壁缺陷结核分支杆菌对实验动物质量控制影响的研究

为了解细胞壁缺陷结核分支杆菌对豚鼠的致病作用 ,探讨细胞壁缺陷结核分支杆菌对实验动物微生物质量控制的影响 ,将结核分支杆菌稳定 L型 ( TBL )纯培养物注射 PPD试验阴性豚鼠腹股沟皮下 ,检测结核分支杆菌及其 L型和观察动物的发病情况、PPD反应、组织病理学改变。结果表明 ,L型感染的动物中 5× 10 8个/ m L 剂量感染组 14～ 18d后可见局部皮肤红肿及腹股沟淋巴结肿大 ,但各组动物至第 4周PPD试验仍为阴性。细菌学分离培养未发现结核分支杆菌 ,组织的病理学检查可见结核的典型和非典型增生现象。采用非高渗培养法或结核特异性 PCR反应 ,能够从结核分支杆菌稳定L 型感染动物的组织标本内获得结核分支杆菌稳定 L型感染的阳性结果     

Use of a Mycobacterial Cell Wall Extract (MCWE) in Susceptible Mares to Clear Experimentally Induced Endometritis With Streptococcus zooepidemicus

The ability of an immunomodulator, mycobacterial cell wall extract (MCWE), to clear uterine infection in susceptible mares after an experimental challenge withStreptococcus zooepidemicus was evaluated. Thirty mares susceptible to endometritis, based on the presence of uterine fluid during both diestrus and estrus, were selected from a herd of 896 and inoculated with a live culture of 5 × 10⁶ CFU of S. zooepidemicus on day 1 of estrus. Twenty-four hours later, mares were evaluated by ultrasonography, bacteriology, exfoliative cytology, and uterine biopsy to confirm infection. Forty-eight hours after inoculation, and on confirmation of uterine infection, mares were randomly assigned to one of four unbalanced experimental treatments to receive 1500 μg MCWE IU (n = 10) or IV (n = 10), or placebo IU (n = 5) or IV (n = 5). Mares were examined at ovulation and 7 days post-ovulation for uterine fluid via transrectal ultrasonography and for bacteriology, exfoliative cytology, and uterine biopsy. Efficacy was based on the ability of the mare to clear endometritis as determined by negative bacteriology and reduced numbers of polymorphonuclear cells (PMNs) on uterine biopsy. Because no statistical difference was detected between routes of administration on day 7 post-ovulation, the data sets were combined and re-analyzed to evaluate overall efficacy. Endometritis was observed in all placebo-treated mares 7 days post-ovulation, whereas treatment with MCWE resulted in the elimination of endometritis in 35% of the mares by the time of ovulation, and 70% of the mares by 7 days post-ovulation. Treatment with MCWE, compared with the placebo group, resulted in a significant decrease in the number of mares positive for endometritis at ovulation based on exfoliative cytology and bacteriology (P < .01) and at 7 days post-ovulation based on biopsy, exfoliative cytology, and bacteriology (P < .001). Results indicate that MCWE was an effective treatment for the elimination of endometritis caused by S. zooepidemicus in mares.     

Evaluation of Needle-free Injection Devices for Intramuscular Vaccination in Horses

Needle-free injection devices have been approved for the delivery of biologics with inherently low immunogenicity, such as plasmid DNA vaccines; however, no studies have described their use in equine patients. This article compares the use of two such devices (VitaJet-3 and Biojector2000) at typical vaccination sites in a cohort of six horses. After identifying the optimal device and vaccination site, a second cohort of five horses was used to document the biologic activity of a DNA plasmid vector delivered with the selected injector. Injector characteristics, including the amount of intramuscular drug deposition, residual skin dose, and pain responses, were evaluated following vaccination, with colored saline in the pectoral muscles and cervical region in six horses. The optimal device was then selected and used for intramuscular vaccination with the pING/tyrosinase plasmid vector in a group of five horses. Biological activity was measured through antibody response to the protein encoded by the plasmid on days 0, 14, 28, 42, and 56 postvaccination. Optimal intramuscular dose delivery was obtained in the pectoral muscle site using the VitaJet-3. No significant pain responses were noted. Dependent edema was seen at vaccination sites 24 hours after therapy. Antibody responses to the protein encoded by the DNA plasmid vector significantly increased after vaccinations in all horses. The VitaJet-3 is easy to use and is effective for delivering intramuscular vaccinations with DNA plasmid vectors in horses. This device allows for vaccination with vectors that exhibit low immunogenicity and/or that require targeted delivery to specific tissue planes.     

The Study of Cell Wall Breaking Methods to Obtain Maximum Extraction Rate of Yeast Polysaccharides

The aim of this study was to determine the optimal method of the yeast cell wall breaking which could obtain the maximum extraction rate of yeast polysaccharide. The extraction rate of yeast polysaccharide and the cell wall broken rate were used as the evaluating indexes. The conditions of ultrasonic disruption, repeated freezing and thawing, ultrasonic cleaning and protease treatment were optimized to select the appropriate parameters in cell wall breaking. The effects of these 4 methods and the synergistic action of ultrasonic disruption and protease treatment were compared. The results showed that when the ultrasonic cleaning time was 40 min and ultrasonic disruption time was 50 min, the water content of slurry was 15% and the times of freezing and thawing was 3, and the protease treatment was carried out at 40℃ for 15 h with enzyme concentration 300 IU/g and pH 5.0, respectively, the extraction rates of yeast polysaccharides were the highest. The effect of protease treatment was the best among these 4 cell wall breaking methods, its extraction rate of polysaccharide was 149.81 mg/g and the cell wall broken rate was 53.49%. The effect of combined way was better than that of single treatment, but the synergistic action would destroy the integrity of polysaccharide's structure. So the protease treatment was thought an ideal cell wall breaking method of yeast.     

获得最大酵母多糖提取率的破壁方法研究

试验旨在选择获得最大酵母多糖提取率的酵母菌破壁方法。以酵母多糖提取率与细胞破壁率为综合评定指标,首先对超声波破碎、超声波清洗、反复冻融、蛋白酶处理4种方法进行条件优化,选择最适破壁条件;然后将此4种单一的破壁方法与超声波破碎和蛋白酶处理协同方法相比较,确定获得最大多糖提取率的破壁方法。结果表明,当超声波清洗时间达到40 min、超声波破碎时间达到50 min时,酵母多糖提取率达到最大值;当菌泥加水量和冻融次数分别为15%与3次时,多糖提取率最大;当蛋白酶用量300 IU/g、酶解时间15 h、pH 5.0、处理温度40℃时,蛋白酶处理的多糖提取率最高。4种破壁方法中蛋白酶处理的效果最佳,对应的多糖提取率为149.81 mg/g,破壁率为53.49%。协同破壁处理的提取效果要优于单一的破壁方法,但考虑到协同作用对多糖结构的完整性构成威胁,将蛋白酶处理确定为最佳的酵母菌破壁方法。     

The Effect of Mycobacterium Cell Wall Fraction on Histologic,Immunologic, and Clinical Parameters of Postpartum Involution in the Mare

Maintaining yearly foal production is important for the economic success of the broodmare, and this requires breeding to occur as quickly postpartum as possible. The initial postpartum estrus occurs within 5–20 days postpartum, whereas the uterus is still undergoing repair from tissue alterations during pregnancy and parturition, a process known as involution. Attempts have been made to hasten this process, but with minimal success. Mycobacterium cell wall fraction (MCWF) is an immunomodulator that has been shown to reduce bacterial growth and alter aspects of the immune response to breeding, but it is unknown if MCWF hastens the process of involution. Therefore, the objectives of this study were to (1) investigate the effect of MCWF on tissue remodeling, (2) assess the effect of MCWF on the local immune system of the uterus, and (3) determine the optimal treatment interval needed for these processes to occur. We hypothesize that repeated treatments of MCWF postpartum will hasten the process of involution. To study this, 16 pregnant mares of mixed breeds were evaluated postpartum. Control mares (n = 4) received 1.5 mL lactated Ringer’s solution intravenously on Day 1 (Day 0 = day of parturition) postpartum and again on Day 7, whereas treated mares either received 1.5 mL Settle intravenously on Day 1 and Day 7 (TX1; n = 6) or 1.5 mL Settle intravenously on Day 1 and then every 3 days until ovulation was detected (TX2; n = 6) and then evaluated until 15 days postpartum. Mares were assessed every 3 days for clinical, immunologic, and histologic parameters. Clinical parameters were assessed with transrectal ultrasonography and included ovarian activity, uterine fluid retention, and measurement of the uterine diameter, in addition to endometrial culture. Immunologic parameters included endometrial biopsies for quantitative polymerase chain reaction for expression of various cytokines (interleukin [IL]-1β, IL-1RN, IL-4, IL-6, IL-8, IL-10, tumor necrosis factor [TNF], interferon [IFN]-γ, and granulocyte-macrophage colony-stimulating factor) in addition to endometrial cytology. Formalin-fixed endometrial biopsies were histologically assessed for the retention of microcaruncles, dilation of endometrial glands, and inflammation of the mucosa, stratum compactum, and spongiosum. Statistics were performed using SAS 9.4, using a mixed model for repeated measures with mare and treatment as a random effect. All post-hoc analysis was done using a Tukey’s honestly significant difference test. Involution was considered complete by Day 15 postpartum in all mares, and the day postpartum had a significant effect on almost all parameters investigated, indicating the immunologic process of involution. Treatment with MCWF decreased the magnitude of bacterial growth in addition to time to negative culture. In addition, MCWF increased the expression of IL-1β, IFNγ, and TNF. Although minimal treatment effect was noted histologically, a decrease in mucosal inflammation was seen in MCWF-treated mares. In conclusion, involution appears to be influenced by the immune system. In addition, MCWF appears to have a bactericidal effect on the postpartum mare, and this may be because of an increase in proinflammatory cytokines. It is unknown if this bactericidal property will improve fertility on the first estrous cycle postpartum, and future studies are needed to determine this.     

金黄色葡萄球菌细胞壁变化与β-内酰胺类抗生素耐药的关系

旨在揭示金黄色葡萄球菌针对β-内酰胺类抗生素可能存在细胞壁增厚的耐药机制。2016-2018年间，采集宁夏地区部分奶牛养殖场临床及亚临床型乳腺炎的乳样，通过显色培养基鉴别、镜检及PCR方法，分离鉴定牛乳源金黄色葡萄球菌；利用微量肉汤稀释法测定细菌对14种抗菌药物的耐药性，了解本地区金黄色葡萄球菌的耐药率及多重耐药情况；通过qRT-PCR方法检测细胞壁增厚相关的pbpB、murG、glmU、atlR基因转录丰度，并结合透射电镜进行形态观察，以确定增厚及发生原因。结果显示，分离鉴定出261株牛乳源金黄色葡萄球菌，其中包括9株耐甲氧西林金黄色葡萄球菌（methicillin-resistant Staphylococcus aureus，MRSA）。药敏试验结果显示，金黄色葡萄球菌对β-内酰胺类抗生素具有较高的耐药率，其中氨苄西林为79.69%，青霉素为78.54%。多重耐药情况是以3、7和8重耐药的菌株居多；其中1株耐药种数达14种之多。qRT-PCR结果表明，4种相关基因的转录丰度均极显著上调（P<0.001或P<0.01）。透射电镜观察发现，甲氧西林敏感的金黄色葡萄球菌（methicillin sensitive Staphylococcus aureus，MSSA）JY21菌株的细胞壁在64和128 μg·mL^-1的青霉素浓度下，较对照组均极显著增厚（P<0.001），并可见细胞壁表面粗糙，有结节状凸起；但药物浓度从64 μg·mL^-1升高至128 μg·mL^-1细胞壁不再显著增厚（P>0.05）。MRSA WLD10菌株细胞壁未出现明显增厚（P>0.05）。综上所述，本地区牛乳源金黄色葡萄球菌针对β-内酰胺类抗生素，存在细胞壁增厚的耐药机制；增厚的原因主要是肽聚糖的过度合成及细胞自溶的减少。与MSSA JY21菌株相比，细胞壁增厚并非MRSA WLD10重要的耐药机制。     

酵母壁多糖对断奶仔猪外周血免疫和肠道免疫的影响

本试验旨在探讨饲粮中添加酵母壁多糖对断奶仔猪外周血免疫和肠道免疫的影响。采用单因素试验设计方法,选取21日龄遗传胎次、体重接近的断奶仔猪180头,随机分为4个组,每组5个重复,每个重复9头猪。4组分别饲喂饲粮中添加0(对照组)、0.15%、0.30%和0.45%酵母壁多糖的试验饲粮。试验期21 d。结果表明:1)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著提高了断奶仔猪血清免疫球蛋白A(IgA)含量(P0.05),饲粮中添加0.30%和0.45%酵母壁多糖显著提高了断奶仔猪血清免疫球蛋白G(IgG)含量(P0.05)。2)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著降低了断奶仔猪血清干扰素-γ(IFN-γ)含量(P0.05),饲粮中添加0.30%酵母壁多糖显著提高了断奶仔猪血清白细胞介素-10(IL-10)的含量(P0.05)。饲粮中添加酵母壁多糖对断奶仔猪血清白细胞介素-2(IL-2)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量均无显著影响(P0.05)。3)与对照组相比,饲粮中添加0.30%和0.45%酵母壁多糖显著提高了断奶仔猪回肠CD4~+淋巴细胞含量(P0.05),饲粮中添加酵母壁多糖能一定程度提高断奶仔猪回肠CD8~+和CD20~+淋巴细胞含量,但差异不显著(P0.05)。由此可知,酵母壁多糖能一定程度提高断奶仔猪外周血免疫和肠道免疫,缓解断奶应激。     

调节性T细胞及在结核分支杆菌感染中的作用研究进展

结核病是由结核分支杆菌引起的慢性消耗性传染病。结核分支杆菌属于胞内寄生菌,机体的抗结核免疫主要依赖特异性的细胞免疫。调节性 T 细胞（Treg）是一类具有免疫抑制功能的 T 细胞亚群,在结核病的发生发展过程中起重要作用,Treg 在抗结核免疫中的作用成为研究热点。论文就调节性 T 细胞在结核分支杆菌感染中作用的研究进展做一综述。     

白色瘤胃球菌对秸秆细胞壁和微晶纤维素的附着比较

本文采用比浊法测定了白色瘤胃球菌 (R .albus 7)对球磨、未球磨玉米秸秆细胞壁和微晶纤维素的附着率。结果表明 ,白色瘤胃球菌对球磨玉米秸细胞壁的附着能力与微晶纤维素相当 ,且二者显著高于未球磨玉米秸细胞壁的附着率 (P <0 .0 1) ,说明增加细胞壁颗粒表面积可以提高纤维分解菌的附着程度。白色瘤胃球菌经酶 (胰蛋白酶、蛋白酶 )和高碘酸钠修饰后 ,可显著降低其对玉米秸细胞壁和微晶纤维素的附着能力 (P <0 .0 1) ,而用甲醛或戊二醛固定细菌蛋白质对细菌的附着能力没有显著影响 ,推测细胞表面的蛋白质和碳水化合物同时参与了对植物细胞壁的附着。细菌经氯化锂修饰后附着能力下降最为明显 ,说明S层蛋白质 (单一蛋白质或糖蛋白 )与白色瘤胃球菌的附着能力有很大关系     

Bilateral Pinnal Squamous Cell Carcinoma in a Dog with Chronic Otitis Externa

Abstract— An aged dog with a chronic, bilateral, bacterial otitis externa developed bilateral squamous cell carcinomas of the pinna. The tumours were located where the pinna would cover the external auditory meatus and probably were induced by the chronic infection.
Résumé— Un chien agé souftrant d'otite externe bactérienne chronique bilatérale présenta un épithelioma spinocellulaire bilatéral des pavilions auriculaires. Les tumeurs étaient situées là où les pavilions auraient recourvert le méat auriculaire et furent sans doute provoqués par l'infection chronique.
Zusammenfassung— Ein alter Hund mit einer chronischen, bilateralen, bakteriell bedingten Otitis externa entwickelte Plattenepithelkarzinome en beiden Ohrmuscheln. Diese Tumore traten an der Pinna exakt an der Mündung des äußeren Gehörgangs auf, und sind möglicherweise durch die chronische Infektion induziert worden.
Resumen  Un perro de edad avanzada que presentaba una otitis externa bilateral bacteriana crónica desarrolló unos carcinomas de células escamosas bilaterales del pabellón auditivo. Los tumores estaban localizados en la entrada del meato auditivo y probablemente fueron inducidos por la infección crónica.     

酵母壁多糖对断奶仔猪肠道挥发性脂肪酸和微生物菌群的影响

本研究旨在探讨饲粮中添加酵母壁多糖对断奶仔猪肠道挥发性脂肪酸和微生物菌群的影响。试验采用单因素试验设计方法,选取180头遗传背景一致、健康状况良好、胎次和体重接近的21日龄断奶仔猪,随机分为4个组,每组5个重复,每个重复9头猪。4个组试验猪分别饲喂对照饲粮(未添加酵母壁多糖)、0.15%酵母壁多糖饲粮、0.30%酵母壁多糖饲粮和0.45%酵母壁多糖饲粮。试验期21 d。结果表明:1)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著提高了仔猪结肠乙酸的含量(P0.05);其中,0.30%和0.45%酵母壁多糖还显著提高了结肠丙酸、丁酸以及总挥发性脂肪酸的含量(P0.05),且二者之间差异不显著(P0.05)。2)与对照组相比,饲粮中添加0.15%、0.30%和0.45%酵母壁多糖显著降低了盲肠沙门氏菌和大肠杆菌数量(P0.05),且0.30%和0.45%酵母壁多糖组之间差异不显著(P0.05)。由此可知,酵母壁多糖可提高仔猪肠道挥发性脂肪酸含量,并改善肠道微生物菌群结构,根据回归方程预测,酵母壁多糖在仔猪饲粮中的适宜添加水平为0.31%~0.40%。     

酵母壁多糖对断奶仔猪生长性能和小肠黏膜形态结构的影响

本研究旨在探讨饲粮中添加酵母壁多糖对断奶仔猪生长性能和小肠黏膜形态结构的影响。试验采用单因素试验设计方法,选取180头遗传背景一致、健康状况良好、胎次和体重接近的21日龄断奶仔猪,随机分为4个组,每组5个重复,每个重复9头猪。4个组分别饲喂对照饲粮、0.15%酵母壁多糖饲粮、0.30%酵母壁多糖饲粮和0.45%酵母壁多糖饲粮。试验期21 d。结果表明:1)与对照组相比,饲粮中添加酵母壁多糖能够显著提高断奶仔猪平均日增重和平均日采食量(P0.05);酵母壁多糖有降低断奶仔猪腹泻率和料重比的趋势,但差异不显著(P0.05)。2)与对照组相比,饲粮中添加酵母壁多糖能够显著提高断奶仔猪空肠绒毛高度/隐窝深度值(P0.05),添加0.45%酵母壁多糖能够显著提高断奶仔猪空肠绒毛高度和十二指肠绒毛高度/隐窝深度值(P0.05)。由此可知,酵母壁多糖可提高断奶仔猪的生长性能,并改善小肠黏膜形态结构;综合生长性能、小肠黏膜形态及经济成本等指标,其在仔猪饲粮中的适宜添加量为0.30%。     

酵母细胞壁多糖提取纯化鉴定技术及其应用研究进展

酵母细胞壁多糖的主要活性成分是β-葡聚糖和甘露聚糖,在动物机体内与肠道受体竞争吸附病原菌以及在肠道降解提供能量,改善动物肠道环境;也会利用其结构与Fe²⁺离子螯合抑制羟自由基的产生以及防止脂质过氧化连锁反应等,提高抗氧化能力;其特有的化学位点会通过多种分子间作用力吸附霉菌毒素;β-葡聚糖和甘露聚糖通过作用于巨噬细胞调节多种细胞因子,通过多个途径增强机体的特异性和非特异性免疫力。目前在养殖业中酵母细胞壁多糖可用于替代金霉素、硫酸黏杆菌素等抗生素控制鸡的坏死性肠炎、减少水产动物细菌感染、降低猪、牛等幼畜腹泻率、预防各种老化性疾病以及促进动物生长发育。对酵母细胞壁多糖现有的提取方法进行比较,提取酵母细胞壁β-葡聚糖,选用超声辅助酶解产率最高、效果最好;用低浓度的氢氧化钠（NaOH）溶液提取甘露聚糖以及酶碱法提取酵母细胞壁多糖效果最好。分离纯化技术包括有脱蛋白、脱色、单一多糖的分离,将其分离纯化效果分别比较,除去酵母细胞壁多糖中蛋白质的最佳方法是Sevage法;脱除色素的最佳方法是颗粒活性炭吸附法;单一多糖分离纯化采用DEAE-纤维素A52阴离子交换柱和Sephadex G-100柱联用的方法最佳。酵母细胞壁多糖结构鉴定一般采用高效液相色谱法（HPLC）测定多糖的单糖组成,高效凝胶渗透色谱测定多糖的均匀性和分子质量,傅立叶变换红外光谱（FT-IR）、气相色谱-质谱（GC-MS）和核磁共振（NMR）测定多糖的精细结构。文章通过对酵母细胞壁多糖的作用及应用、提取、分离纯化和鉴定技术进行阐述,为酵母细胞壁多糖作为抗生素替代品在畜牧业生产中的推广应用提供理论依据,为抗生素替代品的研发提供新的思路。     

绵羊放牧方式及强度对围封恢复草场牧草细胞壁成分及营养价值的影响

选取内蒙古自治区典型草原区封育恢复中天然放牧草场,进行毛肉兼用细毛羊的暖季放牧试验.采用2×5完全随机试验设计,设定不同放牧方式（不放牧,连续放牧,四区、五区、六区轮牧）及2种放牧强度（1.0,1.4 hm2/un.sh）,运用酶分析方法分析比较了围封过程中放牧绵羊对封育草场牧草细胞壁成分月动态变化及营养价值的影响.结果表明,整个暖季放牧期内,不同放牧区牧草营养物质含量主要受到放牧方式的显著影响（P＜0.05）.具体表现为,不放牧处理牧草细胞壁有机物（OCW）含量始终显著低于各放牧处理,不同放牧处理间没有明显差异;划区轮牧处理牧草始终保持着较高的消化性部分且显著高于连续放牧处理;不放牧处理牧草始终保持着较高的Oa和较低的Ob,而连续放牧处理与其相反,各轮牧处理间没有显著差异;轮牧处理总可消化养分（TDN）和代谢能（ME）含量较高,尤其在牧草生长旺盛期（八月份）,六区轮牧放牧场牧草可利用营养物质含量最高,营养价值最高.     

Development of Immunologic Assays to Measure Response in Horses Vaccinated with Xenogeneic Plasmid DNA Encoding Human Tyrosinase

Xenogeneic plasmid DNA constructs have been developed and optimized for immunotherapies targeting cancer in both humans and dogs. Specifically, plasmid vectors containing the tumor antigen tyrosinase have demonstrated immunoreactivity and clinical benefit in the treatment of melanocytic tumors in these species. Overexpression of tyrosinase has also been noted in equine melanocytic tumors, supporting its role as a valid tumor antigen in the horse. Vaccination with plasmid constructs containing tyrosinase may thus have translational immunoreactivity in the treatment of equine melanomas. Here, we describe a methodology that is highly sensitive and specific for the detection of both humoral and cell-mediated immunoreactivity against tyrosinase in equine patients. These antigen-specific immunoassays are used to measure the humoral and cell-mediated responses in a cohort of horses vaccinated with xenogeneic plasmid DNA encoding human tyrosinase. Serum humoral responses were measured using standard enzyme-linked immunosorbent assay technique against the full-length recombinant human tyrosinase protein. Peripheral blood mononuclear cells were collected from vaccinated horses and stimulated with tyrosinase-specific peptides. Cell-mediated responses were then measured using a novel quantitative real-time-polymerase chain reaction technique to determine resultant interferon-γ expression. All horses developed significantly positive humoral and cell-mediated immune responses compared with their individual prevaccination values. No adverse reactions or signs of autoimmunity were detected. Vaccination with xenogeneic plasmid DNA expressing tyrosinase appears to elicit tumor antigen-specific reactivity and should be evaluated in a larger cohort of horses with melanocytic tumors.     

Evaluation of the Abbott LCx Mycobacterium Tuberculosis Assay for Direct Detection of Mycobacterium Bovis in Bovine Tissue Samples

The commercial LCx amplification assay, usually employed to detect the Myocobacterium tuberculosis complex in respiratory specimens, was evaluated by comparing the results it gave with those obtained using Löwenstein-Jensen solid medium and pathological findings on 55 lymph nodes from cattle with positive and 10 lymph nodes from cattle with negative skin tests for tuberculosis. Fifty-three cultures (51 and 2, respectively) were positive for M. bovis, while the results for the LCx assay and the histological method were positive in 48 (45, 3) and 24 (20, 4) samples, respectively. None of the samples from cattle from certified tuberculosis-free herds were positive by any of the procedures. The results obtained with the LCx assay, compared with the culture procedure, regarded as the gold standard among the diagnostic techniques, gave a specificity of 91.6% and sensitivity of 90.5%. Although the sensitivity of LCx was suboptimal, DNA of M. bovis was detected in 81.8% of the skin test-positive animals. Amplification techniques could provide a rapid and reasonably reliable tool for detecting bovine tuberculosis.     

11种牛分枝杆菌抗原在牛结核病诊断中的初步评价

为筛选及评价用于牛结核病诊断的抗原,本试验将CFP-10、ESAT-6、TB10.4、TB27.4、MPT51、MPT63、MPT64、MPB70、MPB83、Rv3872和Ag85B共11种牛分枝杆菌抗原分别作为包被抗原建立间接ELISA方法,比较其对牛结核病的检出率;同时利用豚鼠和牛的皮试试验评价重组蛋白作为皮试试验刺激原的潜力。此外,将重组蛋白分别刺激结核病阳性牛和阴性牛的抗凝血24 h,检测血浆中的IFN-γ水平,评价各重组蛋白作为IFN-γ释放试验刺激原的潜力。结果显示,不同重组蛋白对结核病阳性血清的反应活性不一,MPB70总检出率最高,为59.7%;其次是Ag85B、ESAT-6和MPB83,检出率均在45%以上;MPT51的检出率最低,仅为2.2%。豚鼠和牛皮试试验均显示,单个重组蛋白作为刺激原难以产生令人满意的迟发型过敏反应（delayed type hypersensitivity,DTH）,而TB10.4、TB27.4、MPT64、MPT63或Rv3872作为补充抗原,分别与CFP-10或ESAT-6混合,均可特异性地刺激结核病阳性牛产生较强的DTH反应,且与PPD-B无显著差异（P>0.05）。重组蛋白CFP-10、ESAT-6、TB10.4和MPT51均能刺激结核病牛全血释放一定的IFN-γ,其中CFP-10、CFP-10-ESAT-6串联蛋白和MPT51刺激结核病阳性牛全血释放的IFN-γ显著高于阴性牛（P<0.05）。因此,这11种牛分枝杆菌抗原并不适合单独用于牛结核病的血清学诊断、皮试试验或IFN-γ释放试验,但以CFP-10和ESAT-6为核心,TB10.4、TB27.4、MPT64、MPT63、Rv3872或MPT51作为其补充抗原,均能提高检测敏感性,有作为皮试试验和IFN-γ释放试验特异性刺激原用于牛结核病诊断的潜力。     

Comparison of Skin Prick Tests with In Vitro Allergy Tests in the Characterization of Horses with Recurrent Airway Obstruction

We intended to identify relevant immunoallergic factors and to compare skin prick tests (SPTs) and in vitro allergy tests in the characterization of horses with recurrent airway obstruction (RAO), so as to ascertain that SPTs perform better. Forty Lusitano/cross-Lusitano horses (30 RAO cases and 10 healthy control horses)—a very valuable autochthonous breed—were studied. Clinical history, thoracic radiography, respiratory tract endoscopy, and bronchoalveolar lavage were used for diagnosis. Serum samples of all 40 horses and undiluted bronchoalveolar lavage fluid samples of 21 RAO horses and 6 control horses were submitted for evaluation by an allergen-specific immunoglobulin E (IgE) enzyme-linked immunosorbent assay. SPTs were performed on the 40 horses. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated for all diagnostic methods. Agreement between diagnostic methods was assessed by kappa statistic (Κ). Chi-square test with Yates correction showed SPT results from the RAO and control groups to be statistically different (P < .05). SPTs showed higher sensitivity, specificity, positive predictive value, and negative predictive value than both enzyme-linked immunosorbent assay tests. In human medicine, SPTs are considered to be the gold standard of allergy tests. Neither serum IgE nor bronchoalveolar lavage fluid IgE reliably detected allergen hypersensitivity, compared with SPT. SPTs performed significantly better overall than both in vitro tests. Low sensitivity of the in vitro assays indicates the need for continued study to elucidate a more sensitive specific IgE test.     

象草细胞壁成分与干物质体外消化率关系分析

以提高干物质体外消化率(IVDMD)为目标,在保持一定干物质产量的同时,通过研究象草细胞壁成分与消化率之间的关系,确定影响干物质体外消化率的细胞壁成分,为筛选高消化率象草育种材料和选育品种提供依据.结果表明:矮生多蘖、较高中蘖、较高少蘖以及高大少蘖型于物质体外消化率依次增加,细胞壁成分NDF、ADF、ADL、HEM、CEL含量则依次降低;第一次刈割时的叶片、茎鞘干物质体外消化率均明显高于第二次刈割,同一刈割时期的茎鞘干物质体外消化率显著高于叶片.叶片干物质体外消化率与NDF、CEL含量表现为显著或极显著负相关,茎鞘干物质体外消化率与NDF、ADF、CEL、ADL含量以及ADL/HEM、ADL/CEL、ADL/(HEM+CEL)比值之间表现为显著或极显著负相关.对IVDMD与细胞壁成分进行主成分分析可得两个较为稳定的主成分,一为ADL含量及ADL/CEL、ADL/(HEM+CEL)比值,二为HEM含量及HEM/CEL比值,因此认为ADL、HEM含量为象草细胞壁成分中影响干物质体外消化率的的主要因素.