吡喹酮氯仿油制剂治疗山羊日本血吸虫病研究

通过口服吡喹酮氯仿油制剂（25mg/kg）治疗人工感染山羊日本血吸虫病，39天后解剖冲法，计数残存虫体，与对照组相比，减虫率为95.7%，减雌率为96.7%。早期文献报道（1981)，当口服吡喹酮剂量20mg/kg时，可获得99.8%的减虫率和100%的减雌率。鉴于国外已有吡喹酮不能完全治愈病人的报道和血吸虫可能已有抗吡喹酮的抗药性，吡喹酮治疗动物血吸虫的适宜治疗剂量值得商讨。     

吡喹酮新型注射剂在小鼠体内的杀虫效果与虫体的消亡规律

血吸虫病的危害极其严重,现有的防治办法首选是化学治疗,临床应用最多的方式是吡喹酮片剂或粉剂的口服,考虑到给动物投药不便,以及吡喹酮对反刍家畜的应用副作用较多和生物利用率低,研制了吡喹酮注射剂,应用人工感染血吸虫的小鼠,实验吡喹酮注射液的杀虫效果,发现吡喹酮新制剂的治疗达到口服相同剂量吡喹酮同样的减虫效果,     

吡喹酮缓释剂治疗小鼠日本血吸虫病的试验

为了观察吡喹酮缓释剂治疗小鼠日本血吸虫病的效果，本试验以可生物降解的壳聚糖作为高分子包裹材料，制备吡喹酮缓释剂（包封率为56．05％），通过皮下埋植治疗人工感染日本血吸虫尾蚴（60±1条）40d后的ICR小鼠，4周后剖杀小鼠。结果，吡喹酮剂量138．4mg／kg的治疗组减虫率为64．08％（P＜0．01），减雌率75．86％（P＜0．001）；吡喹酮剂量69．2mg／kg治疗组的减虫率为25．29％（P＞0．05），减雌率39．14％（P＞0．05）。毒性试验组小鼠皮下埋植空白壳聚糖微囊后，未见小鼠出现中毒症状和死亡。     

吡喹酮非水溶液注射剂的研制--日本血吸虫病治疗试验

利用本课题组研制的吡喹酮非水溶液注射剂，分别进行了小鼠和牛的日本血吸虫病治疗试验。结果表明，感染日本血吸虫的小鼠按每千克体质量20mg和30mg的剂量肌肉注射吡喹酮的减雌率均达100％，人工感染日本血吸虫的牛按每千克体质量10mg和12mg的剂量肌肉注射吡喹酮的减雌率均达100％，每千克体质量8mg的剂量的减雌率为96．41％。自然感染血吸虫病牛按每千克体质量10mg的剂量肌肉注射后30d，粪便转阴率达90．50％。这一结果说明，研制的吡喹酮注射剂具有良好的治疗效果。     

吡喹酮注射液对水牛血吸虫病的临床治疗效果

采用粪便毛蚴孵化法筛选自然感染血吸虫病水牛,考察30%吡喹酮注射液对水牛血吸虫病的临床治疗效果。在实验性临床试验中,将自然感染血吸虫病水牛随机分为5组,分别为吡喹酮注射液高（20 mg/kg）、中（10 mg/kg）、低（5 mg/kg）剂量组,吡喹酮片组（30 mg/kg）和阳性对照组（不用药组）。给药30 d后,吡喹酮注射液高、中、低剂量组及吡喹酮片组的粪便毛蚴孵化转阴率分别为100.0%、100.0%、77.8%和85.7%。在扩大临床试验中,将自然感染血吸虫病水牛随机分为2组,分别为吡喹酮注射液推荐剂量组（10 mg/kg）和吡喹酮片组（30 mg/kg）,给药30 d后,吡喹酮注射液组的52头患牛的粪便毛蚴孵化转阴率为100%,吡喹酮片组的6头患牛的粪便毛蚴孵化转阴率为100%。研究结果表明,30%吡喹酮注射液对水牛血吸虫病具有良好的治疗效果,给药方便,可以替代传统内服片剂,临床推荐剂量为10 mg/kg。     

IgG1型单克隆抗体和吡喹酮协同预防日本血吸虫病的研究

6组昆明系小鼠人工感染日本血吸虫尾蚴前2小时,组1和组2胃饲低剂量吡喹酮(20mg/kg),并分别腹腔注入鼠抗日本血吸虫IgG_1型单克隆抗体ISj51和ISj55作被动免疫,4天后重复1次,4周后剖杀集虫。结果表明,单抗合并吡喹酮的组1和组2的减雌率分别为45.32％和42.95％,均明显高于(p<0.01)仅作腹腔被动免疫转移ISj 51和ISj 55单抗的组3和组4(分别为15.11％和19.44％),亦高于(p>0.05)仅用单剂吡喹酮的组5(37.41％)。2株单抗在巨噬细胞和嗜酸性粒细胞的介导下,杀伤童虫率接近1:2稀释的8周阳性鼠血清(分别为37.50％,46.88％和48.00％,41.18％)。提示:鼠抗日本血吸虫IgG_1型单抗协同低剂量吡喹酮,具有预防日本血吸虫感染,提高灭雌率,降低吡喹酮毒副作用的优点,对控制和预防日本血吸虫病的传播具有一定的意义。     

透皮给药后吡喹酮在小鼠体内组织分布及药代动力学

目的研究透皮给药后吡喹酮在小鼠体内的组织分布及药代动力学。方法用200mg/mL吡喹酮透皮剂涂擦小鼠腹部后,采用高效液相色谱法检测各时间点小鼠器官(肝、心脏、脾和肾)的吡喹酮浓度,分析其在各器官药代谢动力学特点。结果透皮给药后吡喹酮在小鼠肝脏分布浓度最高,脾和肾次之,心脏的浓度最低。吡喹酮在小鼠肝脏、脾和肾脏c-t曲线,各出现两个峰值,第一个峰值4min,Cmax分别为31.78μg/mL和7.87μg/mL,第二个峰值15min,Cmax分别为27μg/mL,11.1μg/mL。小鼠心脏的c-t曲线,有一个峰值3min,Cmax3.91μg/mL。结论吡喹酮透皮给药后,在小鼠肝脏分布浓度高,达到了临床治疗的要求。     

不同剂量新疆阿魏对血吸虫活性的影响

为了探讨新疆阿魏杀灭血吸虫的功效,分别用新疆阿魏蒸馏萃取物以0.1、0.2、0.3、0.4 g/kg浓度药液灌服治疗感染血吸虫病家兔,每隔3 d灌服1次,4次后停止给药,4周后剖检,门静脉灌注法收集虫体、计算减虫率,并以组织切片检查该药对血吸虫感染兔肝脏的治疗作用。结果显示,减虫率分别为12.30%、17.20%、46.40%、40.80%;经组织切片观察,家兔肝脏肉芽肿明显减小。表明新疆阿魏具有很强的杀灭血吸虫活性的作用,最佳杀虫剂量为0.3 g/kg。     

吡喹酮对雏鸭体内毛毕属吸虫作用的观察

观察雏鸭体内不同发育阶段毛毕属吸虫童虫与成虫对吡喹酮的敏感性。用定量毛毕吸虫尾蚴感染雏鸭,感染2h,1、3、7、12、14、16、25、35d和42d分别一次性灌服500mg/kg吡喹酮。给药后4周解剖雏鸭,采用门静脉灌注法收集虫体,计算减虫率。结果显示,吡喹酮对1、3、7、12、14、16、25、35、42日龄童虫或成虫的减虫率分别为16.9%、18.0%、71.3%、50.2%、36.9%、31.3%、45.3%、58.0%、26.4%,其中以7~35日龄虫对该药最敏感,该结果表明,吡喹酮对雏鸭体内毛毕吸虫不同发育阶段均有杀灭作用,对7~35日龄虫体杀灭效果更为明显。     

日本血吸虫重组IAP蛋白的免疫保护效果评估

为研究日本血吸虫凋亡蛋白抑制因子（inhibitor of apoptosis protein of Schistosoma japonicum,SjIAP）重组蛋白诱导BALB/c小鼠的免疫保护效果,利用PCR技术扩增SjIAP基因,构建重组表达质粒pET-28a（＋）-SjIAP,诱导表达重组SjIAP蛋白,并利用重组蛋白制备兔源多克隆抗体血清。然后,选用SjIAP重组蛋白免疫BALB/c小鼠,利用ELISA检测免疫小鼠血清中的特异性抗体水平,以及免疫小鼠脾脏淋巴细胞在SjIAP重组蛋白刺激后产生的细胞因子水平。强化免疫后,将小鼠进行血吸虫尾蚴攻虫试验,感染38 d,进行剖杀,计算虫体减虫率及肝脏减卵率。Western blot结果表明本研究制备的兔源多抗血清能特异性识别SjIAP重组蛋白。ELISA检测表明免疫SjIAP重组蛋白可诱导较高水平的IgG及IgG亚型（IgG1、IgG2a、IgG2b、IgG3）抗体和IFN-γ、IL-2及IL-4细胞因子。动物试验表明,免疫SjIAP重组蛋白的小鼠与PBS组相比分别获得了31.5%的减虫率和37.2%的肝脏减卵率。免疫SjIAP重组蛋白能诱导小鼠获得一定减虫和减卵保护效果,提示血吸虫凋亡蛋白抑制因子可作为抗血吸虫病的疫苗候选分子。     

Clinical Therapeutic Effect of Praziquantel Injection on Buffalo Schistosomiasis

To observe the clinical therapeutic effect of 30% praziquantel injection on buffalo schistosomiasis, the sick buffalos naturally infected with Schistosoma japonicum were selected by miracidium hatching method. In experimental clinical trials, sick buffalos were randomly divided into five groups, high-dose (20 mg/kg), middle-dose (10 mg/kg) and low-dose (5 mg/kg) praziquantel injection groups, praziquantel tablet group (30 mg/kg) and positive control group. After the treatment of 30 d, the negative conversion rate of miracidium were 100.0%, 100.0%, 77.8% and 85.7%, respectively, in praziquantel injection groups with the high, middle and low dose and oral medication group. In expanded clinical trials, the sick buffalos were randomly divided into two groups, praziquantel injection group (10 mg/kg, i.m.) and praziquantel tablet group (30 mg/kg). The results showed that, the negative conversion rate of miracidium were all 100% in the former 52 patients and the latter 6 patients after 30 d. The research confirmed that praziquantel injection was significantly effective in the treatment of buffalo schistosomiasis and convenient for administration. It was concluded that 30% praziquantel injection could replace the traditional oral praziquantel tablet for the treatment of buffalo schistosomiasis, and the recommended dose was 10 mg/kg.     

Anthelmintic activity of artesunate against Fasciola hepatica in naturally infected sheep

In light of rapidly spreading triclabendazole resistance alternative fasciocidal drugs are urgently needed. Following up on promising results obtained with artemether in Fasciola hepatica infected sheep, we here report the efficacy and safety of artesunate in sheep with a natural F. hepatica infection. Artesunate was administered intravenously and intramuscularly, adverse events were monitored and drug efficacy was elucidated by means of faecal egg and worm burden reductions. A single 40 mg/kg intravenous dose of artesunate induced an egg count reduction of 68.9% and a worm burden reduction of 77.4%. Intramuscular artesunate at 40 mg/kg reduced faecal egg count and worm burden by 97.6% and 91.9%, respectively; whereas at 60 mg/kg it caused 93.2% and 87.1% reduction in faecal egg count and worm burden, respectively. Three sheep died 24-72 h post-treatment with a double dose of 40 mg/kg intramuscular artesunate, showing lethargy, sialorrhoea, reduced rumination and tremors. Egg and worm burden reductions of 93.3% and 83.9%, respectively, were calculated in the three surviving sheep. In conclusion, the interesting fasciocidal properties of artesunate in sheep warrant further investigations with an emphasis on toxicity studies.     

Assessing resistance against macrocyclic lactones in gastro-intestinal nematodes in cattle using the faecal egg count reduction test and the controlled efficacy test

The main objective of the present study was to evaluate the accuracy of the faecal egg count reduction test (FECRT) to assess the resistance status of ivermectin (IVM)-resistant isolates of the cattle nematodes Ostertagia ostertagi and Cooperia oncophora, using the controlled efficacy test (worm counts) as a reference. The second objective was to investigate whether both IVM-resistant isolates showed side-resistance against moxidectin (MOX) under controlled conditions. Thirty male Holstein calves were experimentally infected with 25,000 L3 of an IVM-resistant O. ostertagi isolate and 25,000 L3 of an IVM-resistant C. oncophora isolate. Twenty-eight days later the calves were randomly divided into 2 treatment groups and 1 untreated control group. Animals in groups 1 and 2 received MOX (Cydectin(?) 1%, Pfizer) and IVM (Ivomec(?) 1%, Merial) respectively, by subcutaneous injection at a dose rate of 0.2mg/kg bodyweight. Faecal samples were collected 7 and 14days after treatment and animals were necropsied 14/15days post-treatment. Both the FECRT and the controlled efficacy test demonstrated that the O. ostertagi and C. oncophora isolates were resistant against IVM, with efficacies below 90%. The IVM-resistant O. ostertagia isolate was still susceptible to MOX treatment, as shown by over 99% reduction in egg counts and worm burden. The FECRT suggested borderline resistance against MOX in the IVM-resistant C. oncophora isolate, with egg count reductions between 97% (95% CI: 76; 100) at day 7 and 86% (95% CI: 49; 96) at day 14. However, the controlled efficacy test clearly showed MOX-resistance, with a decrease of only 31% (95% CI: -12; 57) in C. oncophora worm numbers. After MOX treatment, a significantly lower number of eggs per female C. oncophora worms was counted compared to the control group (43% reduction). Due to this reduced fecundity, the FECRT may fail to detect MOX-resistance.     

日本血吸虫还原性辅酶I(SjNADH)基因的克隆表达及其免疫保护效果分析

为了评估日本血吸虫还原性辅酶I(SjNADH)在小鼠体内诱导的免疫保护效果,应用PCR获得SjNADH基因片段,其开放阅读框为474 bp,编码157个氨基酸,并在大肠杆菌中成功表达,纯化得到该重组蛋白。Western blot结果显示,SjNADH在日本血吸虫童虫和成虫中的表达量稳定,SjNADH融合蛋白也能被免疫血清识别,具有较好的免疫原性。应用重组蛋白免疫小鼠能诱导产生较高的特异性抗体水平,并诱导了20.13%的减虫率和23.06%的肝脏减卵率。结果表明,获得的SjNADH重组蛋白在小鼠体内诱导产生了部分免疫保护,有作为血吸虫疫苗候选抗原分子的潜力。     

Efficacy of levamisole pour-on compared with levamisole subcutaneous injection against Dictyocaulus viviparus infection in calves.

The efficacy of levamisole pour-on against Dictyocaulus viviparus was compared to that of subcutaneous levamisole injection. Eighteen calves were raised individually and artifically infected with D. viviparus larvae. Faecal samples were collected 27 and 28 days later and larvae per gram (l.p.g.) determined. The animals were then divided into three comparable groups. Group 1 animals remained untreated as controls. Group 2 animals received levamisole 10% w/v subcutaneous injection at a dose of 5 mg kg-1 and Group 3 received levamisole pour-on 20% w/v at a rate of 10 mg kg-1 applied transdermally. Results of l.p.g. measurements from faecal samples taken 7 and 8 days post-treatment indicated a dramatic reduction in the worm burden of animals in both treatment groups. Necropsies at 14 days post-treatment revealed few adult worms in these groups, indicating a 99 and 98% kill rate for pouron and subcutaneous injection, respectively.     

东方田鼠血清筛选噬菌体展示抗原对小鼠日本血吸虫病免疫效果观察

本研究利用前期研究中东方田鼠血清筛选日本血吸虫成虫噬茵体展示cDNA文库获得的10个阳性克隆中的8个噬茵体克隆单独或联合免疫BALB/c小鼠,观察对日本血吸虫病的免疫预防效果,以筛选其中具有发展成疫苗的候选抗原编码基因.研究发现与空白对照组相比,展示血吸虫锌指蛋白的1号噬茵体克隆免疫组在两次实验中分别获得了32.10%和31.25%的减虫率、61.14%和47.31%的肝脏减卵率,虫体合抱率分别由92.59%、57.39%下降到69.09%、41.03%;其它噬茵体克隆单独免疫组或联合免疫组在第一次实验中均获得了8.02%～32.72%的减虫率和40.19%～69.53%的减卵率,但在第二次实验中未能得到验证.研究结果提示,日本血吸虫环状锌指蛋白(1号克隆)编码基因在疫苗方面具有重要研究价值.