微囊型球虫疫苗的制备及其免疫效果的初步研究
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摘要
鸡球虫病是集约化养鸡业，特别是地面平养肉鸡业最为多见、且危害严重的疾病之一，给养鸡业造成的危害是巨大的。目前防治球虫病的手段主要还是依靠化学药物防治，但由于化学药物防治带来了耐药性、药物残留、环境污染等诸多问题，近年来国内外逐渐加大了对鸡球虫病免疫预防的研究，并取得了一定的成绩。我国在应用球虫疫苗预防鸡球虫病上尚属初期阶段，仍存在有许多问题，尤其是疫苗的剂型少，使用方法过于单一等，难以适应我国复杂的饲养模式和环境。
本研究将微囊技术与免疫学技术结合，研制出一种新型的球虫疫苗剂型即微囊型球虫疫苗。这种新剂型的球虫疫苗包括一丸型微囊球虫疫苗和拌料法免疫用的微囊型球虫疫苗，其最佳的制备条件是：海藻酸钠浓度3％、温度30℃、每次用量25ml，新鲜孢子化卵囊悬液2ml，磁力搅拌器1250r／min搅拌15min，氯化钙浓度5％、反应时间6h，30℃晾干3h。
试验证明：微囊型球虫疫苗用拌料免疫的方法，在明亮、宽敞的环境下进行拌料口服一次免疫的卵囊检出率可达到80％，免疫后5-7天可在免疫小鸡的粪便中检到球虫卵囊。通过拌料口服免疫法，测定微囊型球虫拌料疫苗不同免疫次数的免疫效果，试验结果显示：两次免疫可以获得比一次免疫更好的免疫效果。
分别用强毒、弱毒微囊型球虫疫苗进行免疫效果比对试验，结果显示：使用本地强毒(野毒)苗制成的微囊型球虫疫苗拌料免疫的效果优于外地弱毒苗。
分别用微囊型球虫疫苗(包括一丸型微囊球虫疫苗和拌料法免疫用的微囊型球虫疫苗)和液态滴服疫苗进行免疫预防效果对比试验，结果显示：微囊型球虫疫苗无论是直接灌服免疫还是拌料免疫都能够产生与液态滴服疫苗相似的免疫效果，组间增重差异不显著，但与感染对照组间的差异显著，证明：微囊型一丸口服(灌服)疫苗和微囊型拌料疫苗这两种微囊型球虫疫苗应用于鸡球虫病的免疫预防是可行的，可通过进一步的临床免疫效果试验，获取更多实验数据，为这种新剂型球虫疫苗的推广应用打下基础。
分别将在常温、4℃环境下保存9个月、6个月、2个月、1个月的微囊型球虫疫苗和液态滴服疫苗进行疫苗活性测定试验，结果显示：微囊型球虫疫苗在4℃或常温下保存1-2个月，可以保存疫苗中卵囊的有效活性，且在常温下保存的活性要强于在4℃环境下保存的活性。
本研究研制的新型微囊型球虫疫苗的特点是：微囊型球虫疫苗可以采用拌料免疫的方法，因此能够在进行大规模免疫时大大减少免疫操作的劳动强度，同时，这种微囊型球虫疫苗也可以同目前普遍使用的液状球虫疫苗一样，采用灌服的免疫方法进行免疫，而且采用一丸型微囊疫苗灌服免疫具有剂量更准确、免疫操作技术更简易等特点。制成的固体状微囊型球虫疫苗还具有储存、运输更方便等优点。
A new dosage form of coccidial vaccine was developed successfully in this experiment. The best facture conditions were: the concentration, temperature and dose of sodium alginate were 3%,30℃,25ml respectively. The dose of oocysts was 2ml. The concentration and reaction time of calcium chloride were 5%,6h respectively. The rotating speed and time were 1250r/min,15min respectively. The drying temperature and time were 30℃,3h respectively.
Experiments indicated a high vaccinated ratio of chickens was obtained when taking the coccidial microcapsule vaccine mixed in feed by normal condition.
The efficacy of the coccidial microcapsule vaccine for chickens were detected by mixed the vaccine in feed and orally taken different immunity times. The results indicated that using of the coccidial microcapsule vaccine was convenient and the chickens had strong immunity against coccidiosis after twice immunity.
The efficacy of virulent and attenuated anticoccidial microcapsule form vaccine for chickens were compared. The results showed that microcapsule form of virulent vaccine which prepared by the virulent native field strain could obtained better anticoccidial efficacy than attenuated vaccine.
The efficacy of three forms of coccidial vaccine dosage for chickens were tested and the results indicated it had the similar efficacy between microcapsule vaccine and aqua vaccine.
All of the dosage forms of coccidial vaccines were stored at 4℃and room temperature for 9,6,2 and 1 months respectively, and their activity assay was carried out. The results showed that the coccidial microcapsule vaccine still remain high activity when storing at room temperature for 2 months latter.
本研究将微囊技术与免疫学技术结合，研制出一种新型的球虫疫苗剂型即微囊型球虫疫苗。这种新剂型的球虫疫苗包括一丸型微囊球虫疫苗和拌料法免疫用的微囊型球虫疫苗，其最佳的制备条件是：海藻酸钠浓度3％、温度30℃、每次用量25ml，新鲜孢子化卵囊悬液2ml，磁力搅拌器1250r／min搅拌15min，氯化钙浓度5％、反应时间6h，30℃晾干3h。
试验证明：微囊型球虫疫苗用拌料免疫的方法，在明亮、宽敞的环境下进行拌料口服一次免疫的卵囊检出率可达到80％，免疫后5-7天可在免疫小鸡的粪便中检到球虫卵囊。通过拌料口服免疫法，测定微囊型球虫拌料疫苗不同免疫次数的免疫效果，试验结果显示：两次免疫可以获得比一次免疫更好的免疫效果。
分别用强毒、弱毒微囊型球虫疫苗进行免疫效果比对试验，结果显示：使用本地强毒(野毒)苗制成的微囊型球虫疫苗拌料免疫的效果优于外地弱毒苗。
分别用微囊型球虫疫苗(包括一丸型微囊球虫疫苗和拌料法免疫用的微囊型球虫疫苗)和液态滴服疫苗进行免疫预防效果对比试验，结果显示：微囊型球虫疫苗无论是直接灌服免疫还是拌料免疫都能够产生与液态滴服疫苗相似的免疫效果，组间增重差异不显著，但与感染对照组间的差异显著，证明：微囊型一丸口服(灌服)疫苗和微囊型拌料疫苗这两种微囊型球虫疫苗应用于鸡球虫病的免疫预防是可行的，可通过进一步的临床免疫效果试验，获取更多实验数据，为这种新剂型球虫疫苗的推广应用打下基础。
分别将在常温、4℃环境下保存9个月、6个月、2个月、1个月的微囊型球虫疫苗和液态滴服疫苗进行疫苗活性测定试验，结果显示：微囊型球虫疫苗在4℃或常温下保存1-2个月，可以保存疫苗中卵囊的有效活性，且在常温下保存的活性要强于在4℃环境下保存的活性。
本研究研制的新型微囊型球虫疫苗的特点是：微囊型球虫疫苗可以采用拌料免疫的方法，因此能够在进行大规模免疫时大大减少免疫操作的劳动强度，同时，这种微囊型球虫疫苗也可以同目前普遍使用的液状球虫疫苗一样，采用灌服的免疫方法进行免疫，而且采用一丸型微囊疫苗灌服免疫具有剂量更准确、免疫操作技术更简易等特点。制成的固体状微囊型球虫疫苗还具有储存、运输更方便等优点。
A new dosage form of coccidial vaccine was developed successfully in this experiment. The best facture conditions were: the concentration, temperature and dose of sodium alginate were 3%,30℃,25ml respectively. The dose of oocysts was 2ml. The concentration and reaction time of calcium chloride were 5%,6h respectively. The rotating speed and time were 1250r/min,15min respectively. The drying temperature and time were 30℃,3h respectively.
Experiments indicated a high vaccinated ratio of chickens was obtained when taking the coccidial microcapsule vaccine mixed in feed by normal condition.
The efficacy of the coccidial microcapsule vaccine for chickens were detected by mixed the vaccine in feed and orally taken different immunity times. The results indicated that using of the coccidial microcapsule vaccine was convenient and the chickens had strong immunity against coccidiosis after twice immunity.
The efficacy of virulent and attenuated anticoccidial microcapsule form vaccine for chickens were compared. The results showed that microcapsule form of virulent vaccine which prepared by the virulent native field strain could obtained better anticoccidial efficacy than attenuated vaccine.
The efficacy of three forms of coccidial vaccine dosage for chickens were tested and the results indicated it had the similar efficacy between microcapsule vaccine and aqua vaccine.
All of the dosage forms of coccidial vaccines were stored at 4℃and room temperature for 9,6,2 and 1 months respectively, and their activity assay was carried out. The results showed that the coccidial microcapsule vaccine still remain high activity when storing at room temperature for 2 months latter.
引文
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