心脏体外循环系统检测植物外啡肽活性的研究

本文利用猪胃蛋白酶在37℃条例下降解中盘粉，采集不同反应时间的酶解液样品，低温离心处理后，分别加入蛙心体外循环系统中，测定心肌收缩力变化，并随即加入阿片肽阻断剂-纳洛酮，检测植物外啡肽活性。结果表明，酶解液能够引起心肌收缩力加强，并可被纳洛酮逆转，证明酶解液具有植物外啡肽活性。     

蛋白质过瘤胃保护增强植物外啡肽对山羊生长代谢的调节作用

采用过瘤胃保护技术使饲料中的小麦蛋白避开山羊瘤胃微生物破坏作用,进入皱胃和小肠降解,产生活性肽,增强对山羊生长代谢的调节作用。建立了蛙心体外循环系统,利用外啡肽特异阻断剂—纳洛酮,生物检测不同消化液中植物外啡肽的活性。结果表明:对照期和试验期相比24h采食青草量分别为4 65±0 231,6 413±0 039kg,差异极显著(P≤0 001);血清类胰岛素生长因子 1(IGF 1)分别为153 647±18 670,223 993±19 450ng/ml,差异极显著(P<0 01)。血清生长激素(GH)23 170±0 891,26 792±1 003ng/ml,差异极显著(P<0 01)。血清胰岛素18 103±1 035,23 983±1 600ng/ml,差异极显著(P<0 01)。血清氨基氮0 102±0 003,0 140±0 004mM/ml,差异极显著(P<0 001)。血清氨氮0 206±0 012,0 086±0 006mM/ml,差异极显著(P<0 001)。蛋白过瘤胃保护后的皱胃液以及未保护的瘤胃液能使心肌收缩力显著增强,加入纳洛酮后心肌收缩力都显著性逆转降低,从而证明这些消化液具有较强的外啡肽活性。表明蛋白质过瘤胃保护技术能够有效地增强植物外啡肽对生长代谢的调控作用。     

面筋蛋白外啡肽B5对传染性支气管炎灭活疫苗免疫效果的影响

麦谷蛋白源外啡肽B5来源于麦谷蛋白酶解产物,具有阿片样活性。作者通过进一步优化化学合成工艺制备外啡肽B5,并将其添加到传染性支气管炎病毒(IBV)H120株灭活疫苗中,免疫后21d(35日龄)采用IBV M41株病毒液进行点眼、滴鼻攻毒。通过测定免疫仔鸡的生产性能、淋巴细胞增殖水平、血清抗体水平、临床症状及组织病理学变化等指标,评价外啡肽B5的应用效果。结果显示,经过化学合成工艺优化后获得了高纯度的外啡肽B5(HPLC浓度大于98%),7~35日龄肉鸡的平均体增重试验组比对照组显著提高13.8%,料肉比试验组比对照组显著降低15.1%;同时一定浓度的外啡肽B5能够诱导肉仔鸡产生较强的淋巴细胞增殖反应(P0.01),免疫鸡群抗体水平显著高于对照组,在IBV M41株攻毒试验中,试验组可以显著减轻病毒感染肉仔鸡临床症状,肺和肾无组织病理学变化,免疫保护率达到96.7%。外啡肽B5可望开发成一种新型动物功能性饲料添加剂或动物疫苗佐剂。     

仔猪胃肠道食糜和血液中酪啡肽的检测及阿片样活性分析

将24头仔公猪分为对照组(正常哺乳28 d断奶)、试验I组(添喂β-酪啡肽-7)和试验Ⅱ组(添喂酪蛋白酶解物)。试验组于21 d断奶,再连续添喂酪蛋白胃蛋白酶解液或β-酪啡肽-7 10 d,每天2次(共20mL),至32 d,3组动物各随机取6头同时剖杀,取血、胃食糜和空肠食糜。通过所建立的RP-HPLC方法,在胃食糜和空肠食糜中检测到β-酪啡肽-5、β-酪啡肽-7样物质。对照组、酪啡肽-7组和酪蛋白水解液组,β-酪啡肽-7的平均含量分别为21.12±2.38,23.58±7.49、21.81±9.93μg/g胃食糜和16.99±3.48、19.08±2.85、21.36±4.85μg/g肠食糜;胃食糜中-βCM-7的含量高于空肠食糜,试验组均高于对照组。在各组仔猪的血液中未检测到酪啡肽样物质。肠食糜的细胞生物学试验结果表明:与喂仔猪料的对照组比较,试验I组和试验Ⅱ组对细胞腺苷酸环化酶的活性均具有不同程度的抑制作用,抑制率分别为27.9%和89.0%,抑制作用均能被纳洛酮反转,逆转率分别是43.2%和89.07%。提示在21日龄断奶仔猪饲料中添加酪啡肽或酪蛋白酶解物,其肠道中有酪啡肽释放,阿片总活性升高。     

蛋白发酵产物外啡肽的构成与活性研究

利用人工瘤胃装置,选用经定向培育的瘤胃细菌体外发酵面筋蛋白,并适时提取发酵液进行了植物外啡肽构成与活性的研究。结果显示体外发酵11h,外啡肽的生物学活性最强,肽浓度也相对较高。以超滤结合纸层析法检测外啡肽的构成,所显示的色带表明为10～27肽。人工瘤胃装置的发酵参数pH6.412±0.207、总脱氢酶活性(TDHA)(1.037±0.045)U/mL,D600(细胞菌生长量)1.410±0.065,氨氮(NH3-N)(1318.36±140.68)mg/L,微生物总蛋白(MCP)(6.213±1.014)g/L。采用所制备的植物外啡肽对AA肉鸡肌肉注射不同剂量,同时注射新城疫疫苗,结果显示,试验组新城疫血凝抑制抗体滴度显著性高于对照组,7～35日龄的体增重也显著高于对照组,血清中生长激素(GH)、胰岛素(Ins)、三碘甲腺原氨酸(T3)、四碘甲腺原氨酸(T4)的浓度也显著高于对照组。每周每公斤体质量2mL的外啡肽剂量肌肉注射效果最好。     

双酶联合酶解雄蚕蛾蛋白制备活性肽的工艺条件优化

建立并优化以雄蚕蛾蛋白粉为原料酶解制备活性肽的工艺技术,有助于研发具有功能活性的雄蚕蛾保健产品。首先以雄蚕蛾蛋白的水解度和肽得率为考核目标,对酶解工艺中的酶解液p H、反应温度、反应时间、酶种类及其配比、料液质量浓度及酶用量等因素进行单因素试验。然后采用Box-Benhnken中心组合试验设计法对酶解条件进行优化,建立p H、温度、时间、料液质量浓度等4个因素的酶解效率预测回归模型,并使用Design-Expert 8.0软件对数据做回归分析,验证试验结果提示建立的雄蚕蛾蛋白酶解效率预测回归模型对实测数据有较好的拟合性。最终确定以Alcalase蛋白酶和丹尼斯克碱性蛋白酶对雄蚕蛾蛋白粉进行联合酶解的最佳工艺条件为:酶解液p H 9.5,反应温度55℃,反应时间130 min,料液质量浓度70 g/L。在此优化工艺条件下,雄蚕蛾蛋白粉的水解度为27.32%,肽得率为50.43%,酶解效率综合值为40.47%。     

乳酪蛋白活性肽的开发趋势及应用前景

本文概述了酶解酪蛋白生产生物活性肽的研究现状及开发趋势,提出了当前通过酶解酪蛋白制备活性肽需要进一步解决的问题,并探讨了酪蛋白源活性肽开发趋势。     

米蛋白肽制备工艺研究及产品营养成分分析

本研究以米渣为原料,通过对米渣进行预处理及限制性酶解后,再对酶解液浓缩后喷雾干燥得到米蛋白肽粉,研究其最佳酶解条件及产物性质.结果表明:(1)最佳酶解工艺条件:用酶量为1%,酶解温度为50℃,固液比为1:5,pH值为8,酶解时间3 h;(2)产物米蛋白肽的蛋白质含量为91.5%,分子质量大部分在3000 Da 以下,短肽含量为77.87%,必需氨基酸含量丰富,是一种优质植物蛋白质资源.     

β-酪啡肽-7对断奶仔猪胃酸分泌的影响

本试验旨在探讨β-酪啡肽-7对早期断奶仔猪胃酸分泌的影响及其可能机制。选取刚出生健康仔公猪24头,随机分为对照组和β-酪啡肽-7处理组,其中β-酪啡肽-7处理组仔猪在饲喂与对照组相同日粮的基础上,每天灌胃β-酪啡肽-7溶液。仔猪21日龄时一次性断奶,分别在21日龄和35日龄时屠宰,颈部采血分离血清、取胃肠内容物、胃底组织等,测定胃肠食糜的p H值、胃蛋白酶活性、血清中胃泌素的含量以及胃底组织中H~+-K~+-ATP/Na~+-K~+-ATP酶的活性。结果显示:β-酪啡肽-7能降低断奶仔猪胃内食糜p H,其中显著降低35日龄断奶仔猪的胃内p H值(P0.05),而对十二指肠、空肠以及回肠食糜p H无显著影响;β-酪啡肽-7能够升高21日龄断奶仔猪胃蛋白酶的活性,差异不显著(P0.05),但其能显著上调35日龄仔猪的胃蛋白酶的活性(P0.05);放射免疫分析法检测胃泌素结果显示:灌胃β-酪啡肽-7后仔猪血清中胃泌素含量均高于对照组,21日龄时差异不显著(P0.05),35日龄时极显著升高(P0.05);灌胃β-酪啡肽-7对断奶仔猪H~+-K~+-ATP/Na~+-K~+-ATP酶的活性作用不显著(P0.05)。说明添加β-酪啡肽-7可以有效缓解断奶应激引起的胃酸分泌不足以及胃蛋白酶活性较低等问题,其作用途径可能是通过促进胃泌素分泌实现的。     

伴大豆球蛋白酶解肽抑菌活性研究

建立伴大豆球蛋白酶解产生抑菌肽的最适条件,对酶解产物进行分离纯化后进一步测定其抑菌活性。采用复合风味蛋白酶水解伴大豆球蛋白,测定不同时间的酶解液对大肠杆菌和金黄色葡萄球菌的抑菌活性。结果显示:2 h水解产生的酶解肽具有抑菌活性,其对大肠杆菌和金黄色葡萄球菌的最小抑菌浓度分别是2.6和3.2 mg/mL。2 h水解液经Sephadex-G15分离纯化,采用平板计数法鉴定各组分。结果显示:CP2组分对大肠杆菌具有显著的抑制作用(P<0.05)。本研究证明伴大豆球蛋白的复合风味蛋白酶酶解肽具有一定的抑菌活性。     

Phase I evaluation of CCNU (lomustine) in tumor-bearing cats

1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.     

Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects

Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Diversity of Ectoparasites in Sheep Flocks in Sa~o Paulo,Brazil

The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.     

Evaluation of biohydrogenation rate of canola vs. soya bean seeds as unsaturated fatty acids sources for ruminants in situ

An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.     

Serum biochemical values and mineral element contents of tissues in yaks

Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein     

Evaluation of the Microbiological Status of Milk and Various Structures in Mammary Glands from Naturally Infected Dairy Cows

A knowledge of the microbiological status of milk and of the different structures in the mammary glands has great importance in elucidating the pathogenesis of mammary gland infections. The objective of this study was to evaluate the microbiological status of various structures in the mammary glands from naturally infected dairy cows following slaughter. A total of 94 samples of milk, 184 samples of mammary parenchyma, 168 samples of gland cisterns, and 168 samples of teat cisterns were collected for microbiological examination. Microorganisms were detected in 59.9% of all samples, 67.0% of the milk samples, 70.1% of the mammary parenchymas, 55.9% of the gland cisterns and 48.8% of the teat cistern samples. When all samples were considered, coagulase-negative Staphylococcus were the most prevalent (35.7%) followed by coagulase-positive Staphylococcus (12.2%), Corynebacterium bovis (2.4%), Prototheca sp. (1.9%), and Streptococcus dysgalactiae (1.5%). There was a significantly higher occurrence of microorganisms in the milk and mammary parenchyma compared to the gland cisterns and teat cisterns. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development and survival of free-living stages of gastrointestinal nematodes of sheep and goats on pasture in the Nigerian derived savanna

Four grass plots were sequentially contaminated with goat faeces containing known numbers of unembryonated eggs of predominantly Haemonchus contortus and Trichostrongylus spp. between October 1982 and April 1983. Four other plots were similarly contaminated with sheep faeces between February and May 1987. An additional plot was repeatedly contaminated with sheep faeces from February to April 1987. Populations of free-living stages in faeces and of infective larvae (L₃) in the herbage were subsequently monitored until the end of April and June of 1983 and 1987 respectively. During February and May 1987 two control cultures of sheep faeces were incubated in the laboratory at 25°C–30°C and at a constant temperature of 50°C and the free-living development was also monitored. L₃ developed very readily in the faeces cultured at 25°C–30°C and in those spread on a grass plot in October, at the end of the wet season, but developed less on the plot contaminated in May at the start of the wet season. Worm eggs in faeces deposited on plots during the hot dry season (December to April) or incubated at 50°C died and disintegrated after 24–48 h exposure to the high environmental temperatures. The results indicate that it is unlikely that gastrointestinal nematodes of sheep and goats can develop or survive on open pasture during the dry season in the Nigerian derived savanna zone.