Immunoproteomic analyses of outer membrane antigens of Actinobacillus pleuropneumoniae grown under iron-restricted conditions

Actinobacillus pleuropneumoniae, a bacterial pathogen of swine and agent of porcine pneumonia, causes a highly infectious disease of economic importance in the pig industry. Commercial vaccines for A. pleuropneumoniae include whole-cell bacterins and second generation subunit vaccines but they only confer partial protective immunity. Our search for new vaccine candidates identified antigens that are expressed during conditions that mimic infection; the outer membrane (OM) proteome of A. pleuropneumoniae serotype 5b was examined under iron restriction. Quantitative profiling by 2D-DiGE technology revealed that iron restriction induced expression of previously described transferrin binding proteins (TbpA, TbpB) plus four lipoproteins including spermidine/putrescine binding periplasmic protein 1 precursor (PotD2). Immunoproteomic analyses with antisera from na?ve animals and from infected pigs were able to differentiate antigens within the OM proteome that were specifically recognized during A. pleuropneumoniae infection. Immunoblots of iron-restricted profiles detected PotD2, heme-binding protein A (HbpA), and capsule polysaccharide export protein (CpxD) as well as surface antigens TbpA, TbpB, and OmlA. These data identify OM proteins that demonstrate immunogenicity and upregulation under conditions mimicking infection, providing emphasis on lipoproteins as an important class of antigens to exploit for vaccine development for A. pleuropneumoniae.     

盐胁迫对红豆草幼苗生长和离子积累及分配的影响

以3周龄红豆草(Onobrychis viciaefolia)幼苗为材料,研究了不同浓度(0、5、25、50、100和200mmol·L~(-1))NaCl及50mmol·L~(-1)处理不同时间(0、1、3、5、7和9d)对其生长和离子积累及分配的影响,以期解析红豆草响应盐胁迫的生理机制。结果表明,与对照(0mmol·L~(-1))相比,5、25和50 mmol·L~(-1) NaCl对红豆草幼苗生长影响不大;而100和200mmol·L~(-1)明显抑制其生长。随着盐浓度增加,红豆草地上部和根Na+浓度呈显著增加趋势(P0.05),其中50~200mmol·L~(-1)下,地上部Na+浓度较对照增加14~39倍。特别是200mmol·L~(-1)使根Na+净吸收速率较对照增加了35倍,使整株总Na+量和地上部Na+相对分配比例分别增加了93%和45%;而显著降低了根的K+、Na+选择性运输能力。在50mmol·L~(-1) NaCl下,随着处理时间延长,地上部和根Na+浓度也呈逐渐增加趋势,但根中的Na+到7d时达到最大,然后呈下降趋势;相反,K+浓度均呈缓慢下降趋势,且根中的浓度始终高于地上部。由此可见,在5~50mmol·L~(-1) NaCl下,红豆草通过维持其体内K+、Na+稳态平衡抵御盐胁迫;而在100和200 mmol·L~(-1)下,红豆草地上部积累大量Na+,但其叶片Na+区域化及K+选择性转运能力较弱,导致植株体内Na+、K+稳态紊乱,产生离子毒害,从而抑制其生长。     

猫泛白细胞减少症弱毒冻干疫苗研制与探索最佳冻干曲线的研究

生物制品进行冷冻干燥过程中，或多或少会损失一定的效价，可直接影响产品效力和免疫原性。本文为研制出高效价和保持期长的猪泛白细胞减少症弱毒冻干疫苗，为降低效价损失，选择出最佳冷冻干燥曲线，使冻干后的产品效价损失减少到最低水平。试验结果选用全程为17h的冻干曲线是科学的、可行的。     

颗粒人工饲料育对小蚕生长发育的影响

本文探讨了颗粒人工饲料育的饲养方法和饲育环境对小蚕生长发育的影响，结果表明：1－2龄最适给饵量为50mg/头，适宜饲养密度为1.5头／cm^2，增加给饵次数有利于小蚕的生长发育，温度和光照对小蚕的生长发育有显著的影响，以29－30℃黑暗饲育成绩最好。     

Canine renal failure syndrome in three dogs

Three dead dogs were brought to the College of Veterinary Medicine, Kyungpook National University for study. Clinically, all the dogs showed emaciation, anorexia, depression, hemorrhagic vomiting and diarrhea for 7~10 days before death. All the clinical signs were first noted for about one month after feeding the dogs with commercial diets. At necropsy, all 3 dogs had severe renal damage with the same green-yellowish colored nephroliths in the renal pelvis. They also showed systemic hemorrhage and calcification of several organs, which might have been induced by uremia. Microscopically, necrosis, calcification and calculi were detected in the renal tubules, and especially in the proximal convoluted tubules and collecting ducts of the kidney. These findings were supportive of a mycotoxic effect, and especially on their kidneys. However, the precise cause of the toxic effect in these cases of canine renal failure could not be determined.     

不同氮效率紫花苜蓿叶特征及其产量效应研究

为探讨不同氮效率紫花苜蓿(Medicago sativa)叶特征差异及其对产量的影响,本研究采用田间小区试验,设4个不同氮效率紫花苜蓿品种('LW6010'、'甘农3号'、'甘农7号'和'陇东苜蓿')和2个施氮处理(0,103.5 kg·hm-2),对比研究不同氮效率紫花苜蓿的叶特征及其生产性能,并采用通径分析法探讨叶...     

丝素作为固定化酶载体的研究进展与应用

本文详细描述了制备丝素固定化酶的各种方法，如包埋法、戊二醛共价交联法以及膜状、粉状、线状三种形状的固定化酶的制作；并就固定化葡萄糖氧化酶、过氧化物酶、脲酶、脂肪酶、果胶酶、过氧化氢酶、糖化酶、超氧化物歧化酶、青霉素酰化酶、木瓜蛋白酶、苯丙氨酸裂解酶、β-葡萄糖苷酶等的研究进展与应用进行了介绍和评价。     

Effect of dietary energy source on in vitro substrate utilization and insulin sensitivity of muscle and adipose tissues of Angus and Wagyu steers

Angus (n = 8; 210 kg of BW) and 7/8 Wagyu (n = 8; 174 kg of BW) steers were used to evaluate the effects of dietary energy source on muscle and adipose tissue metabolism and insulin sensitivity. Steers were assigned to either a grain-based (corn) or hay-based (hay) diet and fed to similar final BW. At slaughter, LM and s.c. and i.m. adipose tissue samples were collected. Portions of the LM and adipose tissues were placed immediately in liquid N for later measurement of glycolytic intermediates. Fresh LM and s.c. and i.m. adipose tissues were incubated with [U-(14)C]glucose to assess glucose metabolism in vitro. All in vitro measures were in the presence of 0 or 500 ng/mL of insulin. Also, s.c. and i.m. adipose tissues were incubated with [1-(14)C]acetate to quantify lipid synthesis in vitro. Glucose-6-phosphate and fructose-6-phosphate concentrations were 12.6- and 2.4-fold greater in muscle than in s.c. and i.m. adipose tissues, respectively. Diet did not affect acetate incorporation into fatty acids (P = 0.86). Insulin did not increase conversion of glucose to CO(2), lactate, or total lipid in steers fed hay but caused an increase (per cell) of 97 to 110% in glucose conversion to CO(2), 46 to 54% in glucose conversion to lactate, and 65 to 160% in glucose conversion to total lipid content in adipose tissue from steers fed corn. On a per-cell basis, s.c. adipose tissue had 37% greater glucose oxidation than i.m. adipose (P = 0.04) and 290% greater acetate incorporation into fatty acids than i.m. adipose (P = 0.04). Insulin addition to s.c. adipose tissue from corn-fed steers failed to stimulate glucose incorporation into fatty acids, but exposing i.m. adipose tissue from corn-fed steers to insulin resulted in a 165% increase in glucose incorporation into fatty acids. These results suggest that feeding hay limited both glucose supply and tissue capacity to increase glucose utilization in response to insulin without altering acetate conversion to fatty acids. Because s.c. adipose tissue consistently utilized more acetate and oxidized more glucose than did i.m. adipose, these results suggest that hay-based diets may alter i.m. adipose tissue metabolism with less effect on s.c. adipose tissue.     

Experimental model of swine pneumonic pasteurellosis using crude Actinobacillus pleuropneumoniae cytotoxin and Pasteurella multocida given endobronchially.

This study was designed to develop and characterize a swine pneumonic pasteurellosis model by concurrent introduction of Pasteurella multocida type A and Actinobacillus pleuropneumoniae crude cytotoxin. After a series of preliminary experiments, a combination of 4 x 10(9) P. multocida and 4,000 toxic units of A. pleuropneumoniae crude cytotoxin was determined to produce optimal results. A total of 48 pigs were divided into four groups of 12 pigs each. The control group received buffered saline only. Four pigs from each group were randomly selected for necropsy 3, 7 and 14 days postinoculation (PI). Inoculation of pigs with P. multocida and A. pleuropneumoniae cytotoxin (group 1) resulted in moderate to severe pneumonia. Pasteurella multocida was isolated from pneumonic lesions, grossly normal lung, and bronchial lymph nodes of all group 1 pigs throughout the 14 day experimental period. Pathological changes typical of field cases of swine pneumonic pasteurellosis were produced. Pigs inoculated with P. multocida alone (group 2) had pneumonic lesions and P. multocida was reisolated from lungs at three days PI. Pasteurella multocida was not isolated from these pigs at 7 and 14 days PI, except for one pig in which an abscess developed in the thorax. Pulmonary lesions induced by A. pleuropneumoniae crude cytotoxin alone (group 3) were transient and resolved by seven days PI. Group 1 pigs had significantly greater lung lesion volumes than group 2 and 3 pigs at 3, 7 and 14 days PI. Statistical analysis indicated a significant interactive effect of P. multocida and A. pleuropneumoniae cytotoxin on the development of lung lesion volumes at 7 and 14 days PI (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

奶牛酮病红细胞膜上ATP酶活性变化的研究

为了深入研究酮病的发生机理,选择荷斯坦奶牛作为实验动物,应用酮粉法和改良式水杨醛比色法随机检测分组,Ⅰ组为10头对照牛、Ⅱ组为10 头阳性牛。应用试剂盒比色法测定红细胞膜上ATPase 的活性变化。结果表明,阳性组和对照组相比,红细胞膜上ATPase的活性显著下降(P <0.05)。说明奶牛酮病影响红细胞膜上ATPase的活性变化。