柞蚕蛹血淋巴多酚氧化酶活性影响因素的研究

柞蚕血淋巴中多酚氧化酶活性的大小对柞蚕丝产量有一定影响,并对柞蚕蛹色起着非常重要的作用。本试验采用分光光度计法对柞蚕蛹血淋巴多酚氧化酶活性大小的影响因素进行了研究。结果表明:柞蚕蛹血淋巴多酚氧化酶的最适pH值为5.0;最适温度为25℃;少量的苯基硫脲对其的抑制作用较为明显;不同化合物对多酚氧化酶的活性影响不同,在相同浓度下,氯化钙和氯化镁对柞蚕蛹血淋巴多酚氧化酶的活性有促进作用,其他化合物(EDTA,维生素C和亚硫酸钠等)则表现为抑制作用。     

微粒蒙脱石对猪组织铅水平、红细胞生成和肝脏ALA-D酶活性及铅诱导的脂质过氧化的影响

选60头体质量约33kg的“杜长大”杂交猪，随机分成2组，每组3个重复，一组饲以基础日粮＋10mg/kg铅＋0.5％微粒蒙脱石，另一组饲以基础日粮＋10mg／kg铅作为对照。研究结果表明：添加微粒蒙脱石组与对照组相比，明显降低了猪全血、脑、肝、肾、骨和毛等组织中铅含量：通过血细胞计数、血红素和血球容积测定表明，添加微粒蒙脱石组的红细胞生成显著增高，且肝脏中ALA—D酶活性显著提高。铅对肝脏的损伤作用明显。表现为丙二醛（MDA）含量显著升高（17．08％），过氧化氢酶（CAT）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPx）含量分别降低85．73％、52．17％、47．56％，而在日粮中添加微粒蒙脱石可以显著改善铅诱导的上述损伤。结果提示，微粒蒙脱石对铅中毒有潜在的治疗作用。     

Effect of short‐term supplementation with rumen‐protected fat during the late luteal phase on reproduction and metabolism of ewes

This study was designed to study the effect of short‐term supplementation with rumen‐protected fat during the late luteal phase on reproduction and metabolism of sheep during breeding season. Seventy‐six ewes (Rahmani, Barki and Awassi × Barki) were allocated to two groups considering genotype: the control ewes (C‐group) received a maintenance diet, and the fat‐supplemented ewes (F‐group) received the maintenance diet plus 50 g/head/day of rumen‐ protected fat (Megalac) for 9 days during which oestrus was synchronized. The latter had been accomplished using double intramuscular injection of prostaglandin F_2α (PGF_2α) 11 days apart. Ovarian activity, serum concentration of cholesterol, glucose, insulin and reproductive performance variables were recorded. Data were analysed considering treatment (group) and genotype. Supplementation had positive effects on the overall mean serum concentrations of cholesterol (p < 0.05), glucose (p < 0.05) on day 6 of nutritional treatment and insulin (p = 0.07) on day 8. Fat supplementation did not affect the total number of follicles, follicle populations and ovulation rate. However, fat‐supplemented Rahmani ewes tended to have higher ovulation rate compared with other breeds (treatment × breed interaction, p = 0.06). Treatment also did not affect the mean concentration of serum estradiol or progesterone. Supplemented ewes had higher conception (p = 0.06) and lambing rates (p < 0.05) compared with control. In conclusion, short‐term supplementation with rumen‐protected fat as a source of energy around breeding time improved metabolism, conception and lambing rates of ewes without effects on steroidogenic capacity and ovarian activity being apparent.     

耐酸性α-淀粉酶产生菌发酵条件的优化

试验对从淀粉厂附近的土壤中筛选的产耐酸性α-淀粉酶的菌株发酵条件进行了考察,确定最佳发酵条件为pH 4.5、碳源为可溶性淀粉、氮源为柠檬酸二氢铵、培养温度为35 ℃、接种量为2 mL。      

Trans-10, cis-12 conjugated linoleic acid modulates phagocytic responses of canine peripheral blood polymorphonuclear neutrophilic leukocytes exposed to Clostridium difficile toxin B

Trans-10, cis-12 conjugated linoleic acid (t10c12-CLA) has been reported to enhance phagocyte function. Clostridium difficile toxin B (TcdB) has been known to inhibit Ras-homologous (Rho) guanosine triphosphatases (GTPases) which play essential roles in neutrophil immune functions. Here, we examined whether in vitro treatment with t10c12-CLA modulates the filamentous actin (F-actin) polymerization, phagocytic capacity, and oxidative burst activity (OBA) of canine peripheral blood polymorphonuclear neutrophilic leukocytes (PMNs) exposed to TcdB. Treatment with t10c12-CLA, but not linoleic acid, enhanced PMN F-actin polymerization, phagocytic capacity, and OBA, while TcdB suppressed these functions. t10c12-CLA reversed the suppressive effects of TcdB on these PMN functions. t10c12-CLA stimulated F-actin polymerization regardless of whether phagocytosis was stimulated by microspheres but only elevated OBA when microspheres were added. We asked whether the effects of t10c12-CLA were associated with changes in the activation of the Rho GTPase Cdc42. Treatment with t10c12-CLA augmented Cdc42 activity in both TcdB-treated and TcdB-naive PMNs during phagocytosis. Thus, t10c12-CLA up-regulates PMN phagocytic responses attenuated by TcdB. This effect is associated with an increase in actin polymerization and may involve the activation of Cdc42.     

提取液及固-液分离方法对固态非淀粉多糖酶类活性的影响

本试验旨在探讨固态酶制剂评估中酶的适宜提取液及固-液分离方法。采用4×3双因素完全随机设计,其中提取液分别为去离子水、乙酸-乙酸钠缓冲液(0.1 mol/L,p H 5.50)、磷酸盐缓冲液(0.05 mol/L,p H 6.00)和0.9%Na Cl溶液;溶液提取后的固-液分离方法分别为不分离、3 000 r/min离心3 min和中速滤纸过滤。每个处理5个重复,每个重复设2个平行,测定各个处理下酶的活性,并考察提取液的类型对酶制剂产品(α-半乳糖苷酶除外)溶解离心后溶液中溶质及蛋白质含量的影响。结果表明,磷酸盐缓冲液溶解木聚糖酶后活性最高(P0.05);乙酸-乙酸钠缓冲液、磷酸盐缓冲液及0.9%Na Cl溶液溶解β-葡聚糖酶后活性相当(P0.05),且均显著地高于去离子水(P0.05);去离子水溶解β-甘露聚糖酶后活性最高,其次为乙酸-乙酸钠缓冲液,两者都显著地高于磷酸盐缓冲液和0.9%Na Cl溶液(P0.05)。提取液类型对α-半乳糖苷酶活性无显著影响(P0.05)。酶制剂溶解后的固-液分离方法对木聚糖酶的测定活性无显著性影响(P0.05);提取液离心或过滤后β-葡聚糖酶活性最高(P0.05);提取液离心后β-甘露聚糖酶活性最高(P0.05);而提取液不分离时α-半乳糖苷酶的活性最高(P0.05)。提取液的种类和酶制剂溶解后的固-液分离方法对4种非淀粉多糖酶的测定活性有极显著的交互作用(P0.01)。乙酸-乙酸钠缓冲液对木聚糖酶制剂的溶解度最大(P0.05),去离子水和0.9%Na Cl溶液均对β-葡聚糖酶及β-甘露聚糖酶制剂的溶解度最大(P0.05)。然而,乙酸-乙酸钠缓冲液溶解木聚糖酶、β-葡聚糖酶、β-甘露聚糖酶后提取液中蛋白质的含量均最低(P0.05)。上述结果表明,乙酸-乙酸钠缓冲液溶解4种固态酶制剂可以最有效地将酶蛋白提取出来,α-半乳糖苷酶提取后不宜固液分离,而其他3种酶的提取液适宜进行离心分离。     

猪链球菌2型分泌性核酸酶A(SsnA)活性检测及其基因序列的克隆与测序分析

对8株猪链球菌2型重庆分离株的核酸酶A全基因进行克隆测序,结果表明该基因长度为3 126 bp,与Gen-Bank发表的唯一的该基因的序列相比,核苷酸同源性高于98%,推导的氨基酸同源性高于94%。根据核酸酶A基因的测序结果建立扩增片段长度为301 bp的PCR检测方法,并用甲苯胺蓝DNA酶琼脂对猪链球菌分泌性核酸酶A的活性进行检测。检测了从病猪内脏分离的猪链球菌致病株35株,33株核酸酶A基因PCR检测阳性(阳性比例为94.3%),其中有24株分泌活性检测为阳性(阳性比例68.6%);正常猪扁桃体分离株14株,PCR检测为阳性的10株(阳性比例71.4%),其中有核酸酶A分泌活性为8株(阳性比例57.1%),检测菌株中有2型猪链球菌44株,38株扩增出核酸酶A基因的片段PCR检测阳性并且分泌活性为阳性的有32株,猪链球菌1型、7型、9型、13型、1/2型各1株,均能扩增出核酸酶A基因的片段,但只有13型猪链球菌有核酸酶A分泌活性。对猪链球菌在不同生长时期核酸酶活性的检测显示,猪链球菌在培养4、8、16、32、48 h均有核酸酶A的分泌,并且C a2 和M g2 为分泌性核酸酶A的活性必需因子。