20%吡喹酮注射剂在水牛体内的药代动力学研究

4头成年健康水牛采用随机交叉实验设计,分别进行吡喹酮注射剂肌注和吡喹酮片内服给药的药动学试验.吡喹酮注射剂按10 mg/kg的剂量单次肌注,吡喹酮片按20 mg/kg的剂量单次内服给药.采用高效液相色谱法测定血浆中吡喹酮的质量浓度,方法最低检测限和定量限分别为0.01 mg/L和0.062 5 mg/L.吡喹酮注射剂单剂量肌注给药,血药浓度-时间数据符合一级吸收一室开放模型,其主要动力学参数分别为:t1/2(ka)(0.45±0.029)h,t1/2(ke)(5.04±0.1 0)h,T(peak)(1.72±0.029)h,C(max)(0.87±0.006)mg/L,V(c)(8.58±0.010)L/kg,AUC(7.99±0.005)mg·L-1·h-1.吡喹酮片单剂量口服给药血药浓度-时间数据符合有吸收-室开放模型,其主要动力学参数分别为:Lagtime(0.13±0.010)h,t1/2(ka)(0.76±0.11)h,t1/2(ke),(1.31±0.076)h,T(peak)(3.84±0.026)h,C(max)(0.51±0.006)mg/L,V(c)(26.07±1.221)L/kg,AUC(7.99±0.005)mg·L-11·h-1.肌注相对生物利用度为(232±12.9)%.研究结果表明,20%吡喹酮注射剂肌注给药吸收迅速且完全,具有较高的生物利用度;吡喹酮吸收后在体内广泛分布.     

HPLC法测定兔血浆中吡喹酮的不同前处理方法比较

为了筛选一种合理的吡喹酮(Praziquantel)高效液相色谱分析前处理方法,分别用甲醇、乙酸乙酯、乙醚3种有机溶剂,对相同浓度的吡喹酮血浆样品进行了萃取,并用高效液相色谱法测定其萃取回收率。结果显示:乙酸乙酯和乙醚对吡喹酮的萃取回收率分别在80%和70%左右,甲醇仅为30%左右。结果表明:乙醚和乙酸乙酯可以更好地用于血浆中吡喹酮的萃取。     

水牛口服吡喹酮副反应与处理

目的观察吡喹酮粉剂口服受治水牛出现副反应症状及处理方法的效果，为大面积治疗水牛血吸虫病提供经验。方法试验对受治的4115头水牛进行副反应症状的观察和治疗。结果受治水牛出现腹泻时肌注痢菌净，静注5％葡萄糖液，瘤胃臌气时用口服鱼石脂或大黄苏打片同时皮下注射新斯的明，过敏时肌注地塞米松。结论吡喹酮是用于治疗水牛血吸虫病完全有效的药物，但在服药过程中，受治水牛会出现腹泻＼瘤胃臌气及过敏反应，大部分水牛不经处理副反应会逐渐消失，副反应严重时必须进行对症治疗。     

吡喹酮粉剂和片剂防治水牛血吸虫病效果比较研究

从1994年到1996年，在江西吴城血吸虫病高度流行区研究了3个自然条件相似的自然村共746头血吸虫病水牛，分析了先后用吡喹酮粉剂和片剂进行全面化疗防治水牛血吸虫病的效果。结果显示，吡喹酮粉剂和片剂对革个水平的治疗效果相同；粉剂因在使用时有诸多不便，不易为群众接受，用于化疗普治效果很差，片剂因克服了粉剂的缺．久，使用方便且无副作用，易为群众所接受，用于防治耕牛血吸虫病的化疗普治，效果显著，可促水牛血吸虫病感染率较原来平均下降43．92％，且原感染率越低，下降趋多。     

吡喹酮混悬注射液的质量标准研究

建立吡喹酮混悬注射液的质量标准,为其质量控制提供依据。采用紫外光谱扫描法和高效液相色谱法对吡喹酮混悬注射液进行鉴别。吡喹酮混悬注射液的粒度、分散性、装量、无菌等检查项目按照2010年版《中国兽药典》进行。采用HPLC法测定吡喹酮混悬注射液中吡喹酮含量。结果表明,吡喹酮混悬注射液的各项检查指标均符合国家相关质量要求。建立的含量测定方法简便、可靠、灵敏、重复性好,平均回收率为99.24%,RSD为0.77%,吡喹酮混悬注射液中吡喹酮含量为标示量的99.8%~100.5%。结果表明,吡喹酮混悬注射液质量可靠,质控方法可行。     

RP-HPLC检测复方注射剂中伊维菌素和吡喹酮的含量

为了用高效液相色谱法同时测定复方注射剂中伊维菌素和吡喹酮的含量。采用Hyper-C lone 5u C18柱（250 mm×4.60 mm,5μm）,以乙腈-甲醇-水（53∶35∶12）为流动相,流速为1.0mL/m in,检测波长254 nm,柱温30℃的液相检测方法。结果表明,在此色谱条件下,吡喹酮和伊维菌素的分离良好,吡喹酮和伊维菌素分别在30~500μg/mL和5~40μg/mL浓度范围内峰面积与浓度呈良好的线性关系（r2〉0.99）。按外标法以峰面积计算进行测定,吡喹酮和伊维菌素的平均回收率分别为98.93%和95.89%,RSD分别为0.33%和1.58%（n=9）。说明该方法简便、快速、准确,适用于复方吡喹酮注射液中吡喹酮和伊维菌素含量的测定。     

吡喹酮脂质体的试制及检测

以氢化豆磷脂和胆固醇为膜材,采用逆相薄膜蒸发法制备了吡喹酮脂质体(PZQ-Lip),经电镜和光镜检测显示,PZQ-Lip粒径大部分为2～4 μm,并以大单层脂质体为主.3批PZQ-Lip的平均包封率为(71.82±2.2)%,平均含药量为(8.95±0.2)%.建立的反相高效液相色谱检测PZQ的方法简便、准确,绝对平均回收率按照低、中、高3个浓度水平依次为99.7%(RSD=1.4%,n=5)、99.8%(RSD=1.7%,n=5)和99.5%(RSD=1.8%,n=5).     

吡喹酮注射液的含量测定及稳定性研究

本试验旨在建立吡喹酮注射液中吡喹酮含量的测定方法,并考察其稳定性。采用高效液相色谱(HPLC)法测定吡喹酮注射液中吡喹酮含量。色谱条件:Diamonsil C18柱(150 mm×4.6 mm,5μm),柱温25℃,流动相为乙腈-水(60:40),流速1.0 mL/min,检测波长210 nm,进样量20μL。通过影响因素试验、加速试验及长期稳定性试验考察吡喹酮注射液的稳定性。吡喹酮在6.037~90.555μg/mL范围内与峰面积呈良好的线性关系(R²=0.9993),平均加样回收率为99.24%,RSD为0.77%,吡喹酮注射液的平均含量为标示量的100.3%。在影响因素试验10 d后吡喹酮注射液对高温(60℃)稳定,但对强光照射(4500 lx±500 lx)有一定的敏感性。在加速试验6个月及长期稳定性试验24个月后,吡喹酮注射液的性状、含量等指标均无明显变化。试验结果表明,所建立的含量测定方法简便、可靠、灵敏、重复性好,可用于吡喹酮注射液的质量控制;吡喹酮注射液在室温、避光条件下存放稳定,有效期暂定为2年。     

高效液相色谱法测定赛鸽用复方吡喹酮胶囊中吡喹酮和伊维菌素的含量

用高效液相色谱法同时测定赛鸽用复方吡喹酮胶囊中吡喹酮和伊维菌素的含量.采用Nova-Pak C18柱(150 mm×3.9 mm,4 μm),以乙腈-甲醇-水(53:35:12)为流动相,流速为1.0 mL/min,检测波长254 nm,柱温25℃.在此色谱条件下,吡喹酮和伊维菌素的分离良好,吡喹酮和伊维菌素分别在511～1 534μg/mL和10.1～30.6μg/mL浓度范围内峰面积与浓度呈良好的线性关系(r＞0.999).按外标法以峰面积计算进行测定,吡喹酮和伊维菌素的平均回收率分别为99.9%和98.3%,RSD分别为0.3%和0.6%(n=9).本方法简便、快速、准确,适用于赛鸽用复方吡喹酮胶囊中吡喹酮和伊维菌素含量的测定.     

Effects of species and sex on plasma hormone and metabolite concentrations in crossbred Brahman cattle and crossbred water buffalo

Buffalo meat is tasty and indistinguishable from beef, and has been described as being lean and low in cholesterol in comparison with beef. However, little is known about the plasma hormone and metabolic profiles related to fat metabolism in buffalo, and how their levels are affected by species and sex. In this study levels of plasma hormones and metabolites were compared between species and sexes. Ten growing crossbred Brahman cattle and ten growing crossbred buffaloes (five males and five females in each), with an average age of 22 months were used. The animals were fed on an experimental diet composed of corn silage, brewer's spent grain and a concentrate mixture at a 50:30:20 ratio on a dry matter basis during a fattening period of 6 months. Blood samples were collected every month during the experiment, and were analysed for plasma leptin, insulin, glucose, triglyceride (TG) and total cholesterol (TC) concentrations. Plasma leptin, insulin and TC concentrations increased during fattening and were significantly (P < 0.05) higher in cattle than in buffaloes. These concentrations in cattle showed a big difference between sexes in comparison with buffaloes, and were higher in females than in males. The plasma glucose and TG concentrations did not differ (P > 0.05) between cattle and buffaloes.

The crude fat content of longissimus muscle was significantly higher in cattle than in buffalo. Differences in body fat deposition seem to have an influence on plasma leptin, insulin and TC concentrations differently between cattle and buffaloes.     

Assessment of water buffalo health and productivity in a communal management system in the Philippines

This study aimed to generate a profile of the health and productivity of water buffaloes in a communal setting. Using the Epi-Info version 6.04 for data management, a coded information system was used to accommodate data coming from the reference population. Calves and cows that were born and milked, respectively, were enrolled and monitored for six months. The key outcomes of interest monitored in this study included mortality, morbidity and productivity. Results of the study showed a 93.7 percent probability of the calves surviving up to six months with a calculated mortality true rate of 0.7 deaths per 1000 calf-days at risk. Three calves died during the six month observation period with a mean age at death of 3 days. Analysis of variance on productivity showed that the parasitic load, specifically coccidia, liver fluke and trypanosoma affected the growth rate of the calves. The productivity of cows in the study in terms of milk production was also highly affected by the endoparasitic load and disease condition of the animal. Univariate analysis revealed a significant association between calf scouring and cow’s mastitis (MASTITIS)(P = 0.066). Meanwhile, for the cows, the parasitic load particularly fasciolosis (P = 0.000), coccidiosis (P = 0.002) and trypanosomosis (P = 0.094) (P < 0.10) also significantly affected the milk production. The results give a clearer view of the relationship between the health and productivity profiles of these animals.     

The intravenous glucose tolerance test in water buffalo

OBJECTIVE: The response to intravenous glucose loading in the buffalo using the intravenous glucose tolerance test (IGTT) was investigated to provide a reference for intravenous glucose injection in buffaloes. METHOD: Twelve healthy, fasted, male swamp buffaloes were divided into three groups. Group I: six buffaloes were given 50% glucose at a dosage of 1 g/kg body weight via the jugular vein. Group II: three buffaloes received normal saline. Group III: three buffaloes were not injected. Blood samples were taken from the opposite vein at 60 and 10 min pre-injection (pre60 and pre10), and at 1, 5, 10, 30, 60, 120, 180, 240, 300, 360 and 420 min post-glucose injection (PGI). Plasma glucose was analyzed by the oxidase method. Insulin and glucagon were soon determined with a human radioimmunoassay kit. The insulin (pmol/l)/glucose (mmol/l) ratios (IGR) were also calculated for each sampling time. RESULTS: Mean plasma glucose, insulin and glucagon concentrations of buffaloes in groups II and III were similar at all the sampling times (p > 0.05) and the curves of the IGR for group II and group III were flat throughout. Group I Buffaloes showed an immediate 20 times increase in the mean plasma glucose concentration PGI, over the pre60 and pre10. The peak plasma insulin concentration occurred at 30 min PGI. The mean plasma glucose and insulin concentrations remained above pre-administration levels until 420 min PGI (p < 0.05). However, the mean plasma glucagon concentrations were different only at 1 and 5 min PGI sampling times. The curve of the IGR for group I showed an initial decrease at 1 min PGI, and fluctuated from 10.18 to 25.55 for the remainder of the sampling period. The correlation analysis showed that the mean plasma glucose concentration was positively correlated with insulin level (r = 0.73, p < 0.005), and significantly negatively correlated with mean plasma glucagon (r = -0.58, p < 0.05). The mean plasma insulin level did not show significant correlation with the glucagon (r = 0.06, p > 0.05). CONCLUSION: The hyperglycemia, high insulin, and protracted glucose and insulin curves, the initial decrease in the insulin/glucose ratio indicates that there was an unexpected glucose tolerance to acute intravenous glucose loading in water buffalo compared with other ruminants. The possibly suggested intravenous glucose load in buffaloes is about 5.09-8.28 mmol/l.     

德昌水牛与杂交水牛乳生化组成的分析

对德昌水牛、广西水牛×尼里拉菲水牛F1代乳生化组成进行了分析。结果表明:德昌水牛及F1代乳中蛋白质、乳脂含量和pH较高;SDS-PAGE分析显示德昌水牛及F1代乳蛋白中乳清蛋白比例高,而酪蛋白相对含量低于普通牛乳;德昌水牛乳中的氨基酸含量显著高于杂交水牛和普通牛。     

水牛肉用资源开发与利用

水牛在我国农业与畜牧业的发展历程中占有重要的地位,随着农业机械化的发展,水牛的役用能力被替代。水牛具有耐粗饲、适应强、抗病力强等生理特点,其肉质中蛋白质高、脂肪低,对人类心血管等疾病预防有良好的作用。在我国牛肉市场供不应求的情况下,对水牛肉用资源的合理开发与利用显得尤为重要。本文对我国水牛资源现状、水牛肉用特点与优势等进行了概述,提出了水牛肉用资源开发利用的措施和建议。     

Prevalence of antibodies against Bubaline herpesvirus (BuHV-1) among Mediterranean water buffalo (Bubalus bubalis) with implications in buffalo trade

Background: Both Bovine herpesvirus (BoHV-1) and Bubaline herpesvirus (BuHV-1) have been reported to cross the species barrier. Antibody seroconversion in glycoprotein E (gE) blocking ELISA during BuHV-1 infection has been documented. Recent diagnostic efforts have focused on the development and application of discriminatory tests to distinguish between infections with BoHV-1 and BuHV-1.

Objective: To evaluate the impact and distribution of these two infections in water buffalo farms in two regions (Piedmont (n = 3) and Campania (n = 10), Italy) where infectious bovine rhinotracheitis control programs have been implemented.

Animals and methods: Sampling was carried out on 13 buffalo farms comprising 1089 animals using specific gE-indirect ELISA's test able to discriminate among BoHV-1 and BuHV-1 infections.

Results: 59.0% of animals reacted positive to ELISA (irrespective of whether BoHV-1 or BuHV-1 antigen was used) and 86.4% of these were reactive to BuHV-1 only, whereas 11.8% showed absorbance values for both antigens and were classified as inconclusive. There was a statistically significant age-related difference in BuHV-1 infection rates but not in overall individual (47% vs. 58%) or herd prevalence (100% vs. 90%) of infection between the two regions.

Conclusion: The low percentage of sera reactive to BoHV-1 (1.8%, 12/643) indicates that BuHV-1 may be the main circulating alphaherpesvirus infection in Mediterranean water buffalo in the two study areas. Since Bubalus bubalis is included in Directive 64/432/EEC on animal health problems affecting intra-community trade in bovine animals, diagnostic testing with nonspecific ELISA for BoHV-1 infection in buffalo may yield false-positive reactions. This scenario could lead to economic losses and hamper buffalo trade and movement, particularly for reproduction purposes.     

Seroprevalence and risk factors associated with exposure of water buffalo (Bubalus bubalis) to Neospora caninum in northeast Thailand

Water buffalo are important draft animals for agriculture in resource-restricted areas worldwide. Water buffalo were shown to be experimentally susceptible to infection with Neospora caninum, potentially affected by neosporosis, and naturally exposed to the parasite in Asia. Although enzootic to Thailand, the distribution of N. caninum among Thai water buffalo is unclear. The objectives of this study were to determine the seroprevalence of N. caninum among water buffalo of northeast Thailand and to identify risk factors associated with their exposure to N. caninum. Sera from 628 water buffalo from 288 farms were tested with an indirect fluorescent antibody test (IFAT). A total of 57 samples from 48 herds contained antibodies to N. caninum, indicating overall seroprevalence of 9.1% and 16.7% among individual animals and herds, respectively. The overall seroprevalence was highest in provinces located in the Khorat Basin in the southern part of the region tested. Host age was also associated with seroprevalence, with the greatest seroprevalence (16.1%) among buffalo over 10 years of age, followed by 5–10 years of age (13.4%), 3–5 years (9.2%), and less than 3 years (1.2%). These results collectively suggested that horizontal transmission from canine definitive hosts was an important route of water buffalo exposure to N. caninum. These results also verified the importance of risk factor analysis for effective bovine neosporosis control strategies at the local level.     

A high-performance liquid chromatography method for determination of 4-aminopyridine in tissues and urine

An analytical method was developed to measure 4-aminopyridine in tissues and urine to determine appropriate diagnostic samples in acute poisoning cases. Tissues from rats dosed with 4-aminopyridine were extracted with methylene chloride. Extracts were analyzed by high-performance liquid chromatography using an isocratic solvent system of acetonitrile and aqueous solution (15/85 v/v) consisting of 0.015 M sodium salt of l-heptane-sulfonic acid, 0.002 M tetramethylammonium bromide, and 0.01 M sodium dihydrogen phosphate adjusted to pH 3.0 with phosphoric acid. We concluded that suitable diagnostic samples for acute poisoning cases include stomach contents, kidney, liver, and urine.     

HPLC法检测家兔血浆中地克珠利含量

建立了快速、稳定的家兔血浆中地克珠利提取方法,为进一步研究家兔组织中地克珠利的残留量提供基础。采用HPLC法以妥曲珠利为内标,N,N-2甲基甲酰胺及乙腈联合沉淀法提取家兔血浆中地克珠利,色谱柱为Hypersil ODS（2.5μm,250 mm×4.6 mm）,流动相为乙腈∶水∶三乙胺∶冰乙酸=55∶45∶0.2∶0....     

Quantification of water buffalo (Bubalus bubalis) cytokine expression in response to inactivated foot-and-mouth disease (FMD) vaccine

This study describes the quantification of cytokine expression of vaccinated water buffaloes with FMD inactivated vaccine. Using real-time PCR quantification assay, expression of Th1 (IL-2, IL-12p40, IFNγ); Th2 (IL-4, IL-10) and inflammatory (IL-6, TNFα) cytokines were quantified weekly for the entire three-week duration of the experiment. It was noted that IFNγ, IL-10 and TNFα had peaked on week three post-vaccination while the remaining cytokines peaked on the second week and decreased by the third week. The counteraction between IFNγ and IL-4 was noted as well as the possible suppressive action of IL-10 to that of IL-2 and IL-12, which is a common phenomenon between Th1 and Th2 cytokines. Synergy between TNFa and IL-6 was also observed. These findings suggest that within the immune system of water buffalo there is a dynamic cell-mediated and humoral interaction in response to immunogen. This assessment of the cytokine expressions is vital for the study of water buffalo disease progression and concurring protective immune responses.