贝尼尔聚乳酸微球的制备及体外药物释放

以生物可降解材料聚乳酸（PLA）为载体,用溶剂挥发法的原理制备含贝尼尔（Diminazene Acetrras）的聚乳酸微球。制得的微球平均粒径为（74.82±3.78）μm;贝尼尔含量为（35.53±1.6233）％;微球的收得率为（70.86±4.414）％,贝尼尔的利用率为（50.26±2.1535）％;平均回收率为99.751±1.0917;利用转蓝法研究了微球的体外释药规律,贝尼尔原料药达峰时间为2h,贝尼尔聚乳酸微球的累积释放率在12h时为46.67％,微球无明显突释现象,可缓慢地释药。     

生长激素释放肽-6缓释微球的制备及其对动物生长的影响

以生物降解材料乳酸乙醇酸共聚物(PLGA,LA-GA 8515)为载体,采用复乳-液中蒸发法制备含生长激素释放肽-6的乳酸乙醇酸共聚物微球,并通过体外药物释放试验评价载药微球的释药特征.结果得到收率、粒径大小和分布及含药量均满意的微球,其中包封率为100%,平均体积径为3.45μm,收率为79%.在37℃、pH7.0的生理等渗缓冲液中,首日释药19.19%,其后以平均日释药3.23%的速度释药25 d.小鼠肌肉注射微球30 d后,累积增重比注射生长激素释放肽-6、生理盐水分别高22.56%(P<0.05)和53.17%(P<0.01).结果表明,生长激素释放肽-6乳酸乙醇酸共聚物微球有望发展成为新型长效制剂.     

堆型艾美耳球虫3-1E重组质粒微球的制备及体外释放试验

用聚乳酸羟基乙酸共聚物（PLGA）将鸡堆型艾美耳球虫重组表达质粒pcDNA3.1-3-1E包封,采用水包油包水（W/O/W）双重乳化方法制备pcDNA3-3-1E/PLGA微球。通过正交试验设计优化PLGA微球制备工艺,考察PLGA浓度、聚乙烯醇（PVA）浓度、超声功率、复乳搅拌时间对评价指标（即微球粒径大小、包封率、载药量）的影响,确定制备微球的最佳工艺条件。测定微球的形态、粒径、完整性、质粒DNA包封率、载药量和释放程度;进行体外模拟鸡胃液和肠液试验,观察微球体外释药效果。结果显示,当PLGA浓度为8%、PVA浓度为1.5%、超声功率为60W、复乳搅拌6h为微球制备的最佳工艺参数,在光镜下呈散在圆形,粒径小于12μm。微球的包封率、载药量分别为84.25%和4.46%,裸质粒与微球中质粒超螺旋比例差距不显著,这表明在包被过程中的超螺旋质粒未发生明显的降解。在模拟鸡的胃肠液累积释放试验中,它的累积释放能力依次为pH 3.0〉pH 7.4,载药微球在模拟鸡的胃肠液中24h释放26.8%,模拟肠液中24h释放11.2%,30d时体外累积释放率为81.7%,表明微球具有一定的缓释效果。     

恩诺沙星肺靶向明胶微球的制备及质量评价

研制恩诺沙星肺靶向明胶微球(ENR-GMS)并对其进行质量评价。采用乳化-化学交联法制备恩诺沙星肺靶向明胶微球,油镜下观察微球粒径大小及形态,并用显微照相机(Olym-pusDP12-2)拍照做记录。通过紫外分光光度法检测微球中恩诺沙星(ENR)的含量,进而计算得出其载药量和包封率,并通过溶出度测定法探讨其体外释放规律。制成的恩诺沙星肺靶向明胶微球为淡黄色粉末,油镜下大小较均匀,形态圆整、分散性较好,其内可见被包裹的黄色ENR粉末,平均直径约10.26μm。就粒径分布而言,5～12μm的微球占86.66%,载药量为4.51%,包封率为31%,在pH7.4的PBS缓冲液中的释药符合缓释特征,t1/2约为3.8h,比单纯的ENR延长3个多小时。体外释放实验显示前2.5h时释药量约37.02%,至第5.0h之间释药量达80.87%,之后逐步释放,至10h时药物释放累积量占总药量的87.96%。结果表明,制备出的恩诺沙星明胶微球符合肺靶向给药系统的要求,具有较好的缓释性和肺靶向性。     

富含蛋氨酸醇溶蛋白在产朊假丝酵母中的表达研究

为构建表达富含蛋氨酸醇溶蛋白的产朊假丝酵母工程菌株,利用基因工程技术,将玉米δ—10kD-醇溶蛋白基因（zein）,酵母的GAP启动子（GAP—P）、终止子（GAP—t）拼接合成表达框,以18S rDNA片段为整合介导区．用放线菌酮（CYH）抗性基因作为筛选标记,结合pBR322质粒构建了同源整合表达载体．把zein基因同源重组整合到产朊假丝酵母染色体上,构建表达zein基因的产朊假丝酵母工程菌株。蛋氨酸含量测定结果显示,重组菌株的蛋氨酸表达量比起始菌株提高了17．14％。该产朊假丝酵母工程菌的构建为下一步提高其蛋氨酸产量的研究和应用打下了良好的基础。     

载副猪嗜血杆菌PLGA Omp16蛋白微球的制备及其免疫效果的初步研究

为研究载副猪嗜血杆菌(H. parasuis)外膜蛋白Omp16 PLGA微球的生物学特性,本研究以H. parasuis外膜蛋白Omp16为芯材,聚乳酸-羟基乙酸共聚物(PLGA)为壁材,通过复乳溶剂挥发法制备PLGA微球疫苗。通过扫描电镜测定微球的形态、激光粒径仪测定微球的粒径分布。结果显示,PLGA微球呈球性,表面圆整,粒径分布较窄,平均粒径为5.2μm;采用BCA法测定微球的包封率和载药率,结果显示,微球的包封率为70.5%,载药率为5.78%;以透析释药法测定微球的体外释放特性,结果显示,微球体外释放42 d累计释放抗原95.3%。用该微球0.2 mg/次,免疫小鼠二次(间隔14 d)后,采用ELISA法测定小鼠抗H. parasuis IgG水平,结果显示,微球佐剂组小鼠H. parasuis IgG抗体水平显著高于灭活疫苗组(p0.01);腹腔注射H. parasuis LY02株(血清5型)进行攻毒保护试验,结果显示,微球佐剂组小鼠攻毒保护率(81.8%)显著高于灭活疫苗组(63.6%)(p0.05)。以上试验结果表明,制备的PLGA微球具有良好的缓释性能,能增强小鼠体液免疫应答,提高H. parasuis攻毒保护率。本研究为H. parasuis亚单位微球疫苗的制备奠定基础。     

聚乳酸/丝素共混膜的药物释放性能研究

使用聚乳酸对丝素蛋白膜进行改性，制备聚乳酸/丝素共混膜，期望改善丝素蛋白膜作为药物载体的药物释放效果。首先研究聚乳酸含量对共混膜断裂强力和溶失率的影响，接着以染料罗丹明B作为小分子模型药物，将共混膜作为药物载体，对其药释性能进行研究，并使用药释动力学模型对实验数据进行拟合，最后探讨药物的用量以及膜厚度等因素对共混膜的药释性能影响。结果表明，聚乳酸的加入能够显著地提高丝素蛋白膜的断裂强力，降低其溶失率；聚乳酸/丝素共混膜对供试药物罗丹明B具有较好的缓释性能，释放过程属于菲克扩散模型；适当增加药物罗丹明B的用量可以有效提高共混膜的药物释放速率，而共混膜厚度的增加则可抑制药物的释放速率。     

贝尼尔聚乳酸微球的体外释药研究

试验就贝尼尔聚乳酸微球的体外释药进行了研究，分别考察了相对分子质量不同的聚乳酸、贝尼尔聚乳酸微球中的药物含量不同对体外释药的影响。结果表明：高相对分子质量的聚乳酸微球中的药物释药速度明显慢于低相对分子质量的聚乳酸微球；贝尼尔聚乳酸微球中的药物含量越高，体外释药速度越快。通过体外释药试验的结果可以看出贝尼尔聚乳酸微球具有明显的缓释作用。     

利福平乙基纤维素微球在小鼠体内的组织分布

以有机相分散、溶媒扩散二步法制备了利福平乙基纤维素微球，粒径范围2.5-12.5μm，其中粒径2.5-5.0μm的约占63％。采用微生物测定法研究了利福平乙基纤维素微球静脉给药后，利福平在小鼠体内的组织分布，且与游离利福平在小鼠体内组织分布进行了比较，小鼠单剂量静脉注射游离利福平或利福平乙基纤维素微球（相当于利福平原药）10mg/kg，测定0.5、1、3、7、12、24、48、72、120、168h共10个时间点各组织中的药物浓度。结果表明，游离利福平给药组和利福平乙基纤维素微球给药组在12h内均以肝脏中药物浓度为最高，为9.96-22.41μg/g，在3h内微球给药组高于游离药物给药组，7、12h微球给药组的药物浓度迅速下降，低于游离药物给药组；肾脏的药物浓度在12h内微球给药组明显低于游离药物给药组，24－168h微球给药组的药物浓度下降迟缓，显著高于游离药物给药组直到168药物浓度始终是微球给药组高于游离药物给药组，12h游离药物给药组已不能检出药物含量，而微球给药组直到168h仍维持在5.44μg/g，接近游离药物给药组的最高峰值；血清中利福平浓度在48h已不能检出，而微球给药组在168h为1.33mg/L；心脏、用脏微球给药组和游离药物给药组的药物浓度变化无统计学意义，与游离药物相比，利福平微球明显提高了肺脏中的药物分布，降低了利福平对肝脏、肾脏的毒性，并显著延缓了药物代谢，说明利福平微球具有长效、靶向作用。     

蟾酥总内酯聚乳酸微球的制备、缓释性能及其临床药效评价

为获得具有缓释性能的蟾酥总内酯微球,以75%乙醇及二氯甲烷提取蟾酥粉,获得蟾酥总内酯;以聚乳酸(PLA)为包材,采用乳化-溶剂挥发法制备了蟾酥总内酯PLA微球。以扫描电子显微镜观察微球的形貌及颗粒大小,采用分光光度法测定该微球的包封率及载药量,采用动态透析法考察该微球的体外药物释放过程。采用高效液相色谱-质谱/质谱分析方法,获得了蟾酥总内酯溶液剂及PLA微球皮下注射大鼠后蟾毒灵的药代动力学参数。采用蟾酥总内酯PLA微球分别与泰乐菌素及氟苯尼考配伍,肌肉注射治疗临床猪喘气病,考察微球的协同疗效。结果显示,蟾酥总内酯在蟾酥中的含量为30.1%;制得的蟾酥总内酯PLA微球呈完整的圆球型,大部分微球粒径在1~10μm,微球平均载药量为23.4%,平均包封率为31.5%。在体外微球中蟾酥总内酯24 h释放30%,10 d释放67%,具有明显的药物缓释效果。大鼠皮下注射蟾酥总内酯PLA微球后,蟾毒灵的达峰时间(T_(max))、消除半衰期(t_(1/2))和平均滞留时间(MRT)分别是皮下注射蟾酥总内酯溶液剂的4.0,18.5,2.5倍,微球给药后蟾毒灵的血浆峰浓度值只有溶液剂的23.9%,表明PLA微球具有明显的药物缓释效果,可延长药效作用时间并减少药物毒性。蟾酥总内酯PLA微球与泰乐菌素及氟苯尼考联用治疗临床猪喘气病具有良好的协同作用。     

白术、微米白术和白术多糖对断奶仔猪生长性能和肠道形态及微生态区系的影响

试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。     

Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia

Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.     

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.     

Y‐chromosomal variation of local goat breeds of Turkey close to the domestication centre

Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.     

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.     

Feline intracranial neoplasia: retrospective review of 160 cases (1985-2001)

The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.     

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.