新孢子虫疫苗对山羊免疫效果的初探
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摘要
新孢子虫病(Neosporosis)是由犬新孢子虫(Neospora caninum)寄生于犬、牛、马、羊等多种宿主动物细胞内而引起的一种原虫病。本病呈世界性分布,对牛的危害尤为严重,主要造成孕畜流产、死胎以及新生犊牛的运动神经系统疾病,已给畜牧业生产造成了巨大的经济损失。随着研究的不断深入和研究手段的不断提高,人们利用免疫学、分子生物学和生物技术等先进技术对宿主抗犬新孢子虫的免疫反应及预防犬新孢子虫的疫苗进行了研究,并且取得了喜人的进展。
为了能够更好的了解新孢子虫疫苗的免疫效果,本试验进行了新孢子虫大量培养,并采用反复冻融法提取虫体抗原,辅以佐剂制备新孢子虫疫苗,选择山羊进行了免疫效力试验,采用间接荧光抗体试验(IFAT),对山羊免疫后血清进行了抗体水平监测,测定了山羊妊娠前、后免疫疫苗所产生的不同抗体效价,对山羊接种新孢子虫疫苗后的免疫效果进行监测,为更科学评价新孢子虫疫苗免疫效力奠定了基础。
结果表明,第一组首免两周后,多数山羊产生抗体效价为1：400。二免后抗体水平均显著提高,第六周出现最高抗体效价为1：6 400,攻毒后山羊抗体效价显著增高,最高抗体效价达到1：12 800。
第二组首免后两周大部分山羊均产生抗体,抗体效价约为1：400。二免后所有山羊抗体水平显著提高,第六周左右出现最高抗体效价为1:6 400,攻毒后山羊抗体效价显著升高,产生最高抗体效价为1：12 800,并且六至八周内可维持较高水平。
对照组攻毒后山羊抗体效价迅速升高,攻毒一周后对照组山羊均产生抗体,抗体效价为1：200,攻毒第三周后出现最高抗体效价为1：l 600。
人工感染试验结果表明,第一组在妊娠前免疫,其保护率为71.4%(5/7)；第二组在妊娠后免疫,其第二组疫苗保护率为57.1%(4/7)。第三组为未免疫对照组,其保护率为0,试验结果证明新孢子虫疫苗对母羊流产有较可靠的保护作用。
Neosporosis is a kind of protozoal disease caused by neospora canium parasitizes in various kinds of livestock such as dog, cattle, horse, and sheep. A main reason of cow abortion is neosporosis, which spread widely all over the world, the abortion of cow caused huge economic loses for cattle breeding. Along with the deeply studies and elevated study method, the immunology, molecular biology and biotechnology and other advanced technologies were taken used to study the immunoreaction for host anti-neospora caninum and the vaccines for preventing neospora caninum, many advancements received.
In order to comprehend the immuno-effective of neospora caninum vaccine preferably, a large number of neospora caninum was cultured in this test,then the antigen of neospora caninum was extracted by Freeze+thaw, the neospora caninum vaccine was prepared by mixing the antigen and adjuvant. We took use of sheeps to have the immuno-effective test.IFAT was taken to monitor the antibody level of cow serum after immunization, the different antibody titers were determined, including immunization before gestation and after gestation, evaluating the immuno-effective of vaccine immunization for neospora caninum, which established the foundation for evaluating immuno-effective of vaccine for neospora caninum.
The results showed that the first group, two weeks after the first immunization, the majority of goats to produce antibodies titer 1:400. After second immunization, antibody titer was significantly rised, in the sixth week, the highest antibody titer was 1:6 400, after the artificial infection the goat antibody titer increased suddenly, the highest antibody titer reached 1:12800.
The second group after two weeks of the first immunization, most of the goats would produce antibodies, antibody titer of about 1:400. After second immunization, antibody titer was significantly rised, in the sixth week, the highest antibody titer was 1:6400, after the artificial infection the goat antibody titer increased significant, the highest antibody titer reached 1:12800, and the antibody titer maintain higher between the sixth and eighth weeks after the artifical infection.
With the artificial infection in control group, goat antibody titer increased quickly, after a week the all of the goats in control group had produced antibodies, antibody titer of 1:200, and after the third week, the high titer was 1:1600.
Artificial infection test results show that the first group during pregnancy before the immunization, the protection rate of 71.4%(5/7); the second group after the immunization during pregnancy, the second group of vaccine protection rate of 57.1% (4/7). The third group was not immunized control group, the protection rate was 0, and experimental results show that the neospora caninum vaccine has reliable protection against goat abortions.
为了能够更好的了解新孢子虫疫苗的免疫效果,本试验进行了新孢子虫大量培养,并采用反复冻融法提取虫体抗原,辅以佐剂制备新孢子虫疫苗,选择山羊进行了免疫效力试验,采用间接荧光抗体试验(IFAT),对山羊免疫后血清进行了抗体水平监测,测定了山羊妊娠前、后免疫疫苗所产生的不同抗体效价,对山羊接种新孢子虫疫苗后的免疫效果进行监测,为更科学评价新孢子虫疫苗免疫效力奠定了基础。
结果表明,第一组首免两周后,多数山羊产生抗体效价为1：400。二免后抗体水平均显著提高,第六周出现最高抗体效价为1：6 400,攻毒后山羊抗体效价显著增高,最高抗体效价达到1：12 800。
第二组首免后两周大部分山羊均产生抗体,抗体效价约为1：400。二免后所有山羊抗体水平显著提高,第六周左右出现最高抗体效价为1:6 400,攻毒后山羊抗体效价显著升高,产生最高抗体效价为1：12 800,并且六至八周内可维持较高水平。
对照组攻毒后山羊抗体效价迅速升高,攻毒一周后对照组山羊均产生抗体,抗体效价为1：200,攻毒第三周后出现最高抗体效价为1：l 600。
人工感染试验结果表明,第一组在妊娠前免疫,其保护率为71.4%(5/7)；第二组在妊娠后免疫,其第二组疫苗保护率为57.1%(4/7)。第三组为未免疫对照组,其保护率为0,试验结果证明新孢子虫疫苗对母羊流产有较可靠的保护作用。
Neosporosis is a kind of protozoal disease caused by neospora canium parasitizes in various kinds of livestock such as dog, cattle, horse, and sheep. A main reason of cow abortion is neosporosis, which spread widely all over the world, the abortion of cow caused huge economic loses for cattle breeding. Along with the deeply studies and elevated study method, the immunology, molecular biology and biotechnology and other advanced technologies were taken used to study the immunoreaction for host anti-neospora caninum and the vaccines for preventing neospora caninum, many advancements received.
In order to comprehend the immuno-effective of neospora caninum vaccine preferably, a large number of neospora caninum was cultured in this test,then the antigen of neospora caninum was extracted by Freeze+thaw, the neospora caninum vaccine was prepared by mixing the antigen and adjuvant. We took use of sheeps to have the immuno-effective test.IFAT was taken to monitor the antibody level of cow serum after immunization, the different antibody titers were determined, including immunization before gestation and after gestation, evaluating the immuno-effective of vaccine immunization for neospora caninum, which established the foundation for evaluating immuno-effective of vaccine for neospora caninum.
The results showed that the first group, two weeks after the first immunization, the majority of goats to produce antibodies titer 1:400. After second immunization, antibody titer was significantly rised, in the sixth week, the highest antibody titer was 1:6 400, after the artificial infection the goat antibody titer increased suddenly, the highest antibody titer reached 1:12800.
The second group after two weeks of the first immunization, most of the goats would produce antibodies, antibody titer of about 1:400. After second immunization, antibody titer was significantly rised, in the sixth week, the highest antibody titer was 1:6400, after the artificial infection the goat antibody titer increased significant, the highest antibody titer reached 1:12800, and the antibody titer maintain higher between the sixth and eighth weeks after the artifical infection.
With the artificial infection in control group, goat antibody titer increased quickly, after a week the all of the goats in control group had produced antibodies, antibody titer of 1:200, and after the third week, the high titer was 1:1600.
Artificial infection test results show that the first group during pregnancy before the immunization, the protection rate of 71.4%(5/7); the second group after the immunization during pregnancy, the second group of vaccine protection rate of 57.1% (4/7). The third group was not immunized control group, the protection rate was 0, and experimental results show that the neospora caninum vaccine has reliable protection against goat abortions.
引文
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[2]Perez E, Eonzalezo,Dolz G,et al.First report of bovine neosporosis in dairy cattle in costa Rica[J]. Veterinary Reeord,1998,142(19):520-521.
[3]Hemphill A, Gottstein B.Conraths FJ, et al. A Europcan perspective on Neospora' caninum[J]. International Journal for Parasitology,2000,30:877-924.
[4]Bjcerkas I, Mohn SF. Presthus JZ. Parasitenk,1984,70;271-274.
[5]Dubey JP, Carpenter JL, Speer CA, et al. Newly recognized falal,protozoan disease of dogs:[J]. Vet Med Assoc,1988,192(9):1269-1285.
[6]Dubey JP, Leathers CW, Lindsay Ds. Neospora caninum like protozoon associated with fatal myelitis in new born calves[J]. Parasitol.1989,75:146-148.
[7]Anderson A, Williamson NB. Recent advances in understanding the epidemiology of Neospora caninum in cattle [J]. New Zealand Veterinary Joural,2001,49(2):42-47.
[8]McAllister MM, Dubey JP, Lindsay DS., et al. Dogs are definitive hosts of Neospora caninum[J]. Parasitol,1998,28:1473-1478.
[9]Hemphill A, Muller N, Sager H, et al. Neospora caninum and neosporosis-Basic science at the Institute of Parasitology and possible implications [J]. Schweizer Arch Ⅳ Fur Tierheilkunde,2000,142(5):257-261.
[10]Dubey JP. Neosporosis the first decade of research [J]. Parasitol,1999,29(10): 1485-1488.
[11]Dubey JP, Lindsay DS.Neosporosis [J]. Parasitology Today,1993,9(12):452-458.
[12]Chermette R, Marquer A. Neospora caninum:a new parasite?[J]. Point Veterinaire, 2000,31(208):9-14.
[13]De Meerschman F, Losson B. Neospora caninum:a novel abortive agent in the bovine and-other domestic species[J]. Annales De Medecine Veterinaire,1998,142(5): 299-306.
[14]Brindley PJ, Gazzinelli RT, Denkers EY, et al. Diiferentiation of Toxoplasma gondii from closely related coccidian by fibopfint analysis and'a surface
antigen gene polymerse chain[J]. Trop Med Hyg,1993.48:447-456.
[15]TranasJ, Heinzen RA, Weiss LM, et al. Serological evidence of Human infection with the protozoan Neospora caninum[J]. Clinical and Diagnostic Labomto Immunology, 1999,6:765-767.
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