鸭疫里氏杆菌病灭活疫苗的研制

从河北省某肉鸭场的鸭疫里氏杆菌病发病鸭中分离出鸭疫里氏杆菌（Riemerella anatipestifer,RA)菌株。对该菌株采用液体、固体两种方法增菌培养后制成油乳剂灭活疫苗，以0.5mL/只剂量经皮下免疫7日龄雏鸭，并在河北省某发病肉鸭场进行了田间试验，表明该疫苗免疫效果良好，保护率达90％以上。     

鸭疫里氏杆菌病三价油乳剂灭活疫苗的研究

以1、2、10型鸭疫里氏杆菌（RA）分离株为菌种，研制鸭疫里氏杆菌病三价油乳剂灭活疫苗。经无菌及安全检验合格后，对6日龄樱桃谷雏鸭颈部皮下接种0.4mL／只,免疫后第10、14、21和35d分别用1、2、10型RA强毒株攻攻，第14d的攻毒保护率为91.7%-100%,35d攻毒保护率为66.6%-83.3%。田间试验结果表明，雏鸭5－7日龄免疫后至上市保护率可达95.0%-100%。     

苏中地区鸭疫里氏杆菌血清学调查及多价疫苗的研制

通过玻板凝集试验,对72株鸭疫里氏杆菌(RA)分离株血清型进行了鉴定,血清1、2、3、7和10型分别为11、28、10、7和4株,12株未能鉴定出血清型。选择其中3种优势血清型菌株,制成了三价油乳剂灭活苗,并进行了田间免疫保护试验,结果表明,其对鸭疫里氏杆菌病有较好的预防效果。     

鸭疫里默氏菌油乳剂疫苗的免疫期和保存期的测定

鸭疫里默氏菌(Riemerellaanatipestifer,RA)病主要侵害7～42日龄的各种雏鸭,已成为危害我省养鸭业的主要细菌性传染病。去年,我们已研制出具有较高保护率的鸭疫里默氏菌油乳剂疫苗[1]。为了测定该疫苗的有效免疫期和保存期,我们进行了试验,现报道如下:1　材料与方法　1.1　菌株　系本室自行分离、鉴定、保存的型RA菌株,供制苗及攻菌保护试验用。1.2　疫苗　鸭疫里默氏菌灭活油乳剂疫苗(批号为980811),置于4～8℃保存备用。1.3　试验鸭　取3日龄健康半番鸭200羽,其中180羽用于免疫期测定,20羽用于保存期测定,试验分组情况见表1。表1　　RA…     

鸡EDS—76油乳剂灭活苗的研制及免疫效果观察

以鸡ＥＤＳ－７６ＡＶ－１２７毒株为材料,研制出ＥＤＳ－７６油乳剂灭活疫苗,并对其物理性状、保存期、安全性及免疫效果进行了检测。结果表明：该苗的外观性状、物理稳定性、粘度以及安全性与免疫效果均已达到国内同类疫苗水平;用该疫苗免疫试验鸡,３０ｄＨＩ抗体可达９．６ｌｏｇ２,能很好地抵抗ＥＤＳ－７６ＡＶ－１２７强毒的人工感染;该苗于４℃保存期为１年,２０℃保存期为３个月。找出了油乳剂疫苗水相与油相的最佳合理配比、乳化时间、乳化速度及水相加入油相的速度。     

禽流感二价抗原油乳剂灭活疫苗的研制

以禽流感病毒（ＡＩＶ）Ｈ５Ｎ４株、Ｈ７Ｎ３株为抗原研制了Ｈ５Ｎ４－Ｈ７Ｎ３二价油乳剂灭活疫苗,并对其物理性状、安全性、免疫效力、保存期及抗体消长规律进行了检测。结果表明,试验鸡疫苗在免疫后３周到９个月内对ＡＩＶ－Ｈ５Ｎ４的攻击均获全部保护。疫菌４℃保存１５个月,其免疫效力没有下降。     

黄芪多糖佐剂对Ⅰ型鸭疫里氏杆菌灭活疫苗的作用研究

本研究以Ⅰ型鸭疫里氏杆菌武汉分离株为菌种,经复苏、增菌、浓缩、灭活后与黄芪多糖佐剂混合制成灭活疫苗。试验鸭在5日龄时按最佳免疫剂量（0.3 mL/只）注射疫苗,免疫后第3、7、10、14、21天的攻毒保护率分别为100%、90%、100%、100%、90%,免疫后第3天即能用琼脂扩散法在血清中检测出Ⅰ型鸭疫里氏杆菌特异性抗体,效价逐渐升高,至免疫后第10天达高峰（P<0.01）。试验结果表明,该灭活疫苗能有效预防鸭传染性浆膜炎。     

贵州省鸭疫里默氏杆菌血清2型灭活疫苗制备及免疫研究

为控制贵州省鸭疫里默氏杆菌(RA)的流行,本研究以实验室分离保存的血清2型RA地方优势流行株为菌种,制备了稳定性和安全性较好的甲醛油乳剂灭活疫苗,以其免疫麻鸭后对其抗体滴度进行检测,并于免疫后以RA分离株进行免疫保护攻毒试验。结果显示,该灭活疫苗诱导麻鸭产生的抗体滴度可达1∶3 200,免疫保护率达87.5%,高于商品化的同类灭活疫苗(62.5%)。结果表明利用贵州地区流行的优势血清型RA菌株所制备的疫苗,对防治RA病效果明显。     

鸭疫里氏杆菌二价灭活疫苗制备及免疫效力研究

【目的】根据鸭疫里氏杆菌(Riemerella anatipestifer,RA)血清1型、2型贵州流行株制备二价灭活疫苗,为鸭疫里氏杆菌病的防控及疫苗研制提供研究资料。【方法】以血清1型RA(RA-G06株)、血清2型RA(RA-HS01株)地方流行株为菌种,通过涂板法测定菌株生长曲线,利用改良寇氏法计算菌株对鸭的半数致死量(median lethal dose, LD₅₀),将2株菌培养至终浓度为1×10¹⁰ CFU/mL后等比例混合,以卡波姆为佐剂制备二价灭活疫苗,经疫苗质量检验后进行雏鸭免疫试验;通过检测免疫鸭血清中特异性抗体水平和攻毒保护试验评价疫苗的保护率,对攻毒试验鸭心脏、肝脏、脾脏和脑组织进行组织病理学观察。【结果】RA-G06株和RA-HS01株均在培养12 h时到达峰值,活菌数分别为2.1×10¹¹和3.3×10¹¹ CFU/mL,LD₅₀分别为1.44×10¹⁰和2.63×10⁸ CFU/mL;制备的疫苗安全性良...     

鸭传染性浆膜炎灭活疫苗比较研究

将重庆地区分离鉴定的2株鸭疫里氏杆菌培养、灭活、加入佐剂制成蜂胶、氢氧化铝和油乳剂灭活疫苗，分别免疫5日龄雏鸭，结果表明疫苗是安全的，经一次免疫后10天的保护率分别是70%、50%、75%，15天后的免疫保护率分别是80%、75%、100%；经2次免疫后7天的保护率分别是100%、100%、89%，10天后的保护率均达到100%。     

鸭疫里默氏杆菌贵州流行株蜂胶灭活疫苗制备

【目的】制备鸭疫里默氏杆菌(Riemerella anatipestifer,RA)贵州流行株蜂胶灭活疫苗。【方法】选取RA血清2型贵州流行株(RA-SS-8株)为基础菌株,依次利用分光光度法和平板计数法测定细菌生长曲线,再进行灭活条件筛选,以蜂胶为佐剂制备RA贵州流行株蜂胶灭活疫苗,并进行无菌检验、安全性检验及免疫鸭攻毒保护性试验。【结果】分光光度法测定RA-SS-8株菌液D_{600 nm}值随培养时间变化显示,细菌在0~3 h时增殖缓慢,在3~10 h时增殖趋势明显加快,在10 h以后增殖逐渐趋于平缓,随着培养时间的延长,细菌最终进入衰亡期;平板计数法结果显示,RA-SS-8株D_{600 nm}值为0.1~0.8时,该菌处于对数生长期,且D_{600 nm}值与活菌数呈现良好的线性关系;RA-SS-8株最佳灭活条件为0.2%甲醛溶液、37 ℃灭活12 h;蜂胶灭活疫苗含菌量为3.8×10⁹ CFU/mL,蜂胶干物质含量为10 mg/mL;无菌检验巧克力琼脂培养基上未见菌落生长;安全性检验以2倍免疫剂量接种雏鸭在观察期内未表现出不良反应,大体病变观察未见明显病变;免疫鸭攻毒保护性试验显示,蜂胶灭活疫苗免疫组鸭对RA-SS-8株的攻击后保护率为70%,蜂胶灭活疫苗对试验鸭心脏、肝脏组织均具有良好的保护效果。【结论】本研究成功制备了RA贵州流行株蜂胶灭活疫苗,为蜂胶灭活疫苗制备和动物免疫试验奠定基础。     

Proportions and phenotypic expression of peripheral blood leucocytes in pigs vaccinated with an attenuated C strain and a subunit E2 vaccine against classical swine fever

The influence of an attenuated classical swine fever virus C strain vaccine and a subunit E2 vaccine against classical swine fever on the peripheral blood leucocyte proportion and phenotypic expression in 12-week-old pigs was studied. The C strain was amplified in minipig kidney cell culture and final product contained 10(4 +/- 0.15) TCID50/ml, while the subunit vaccine contained 32 microg per dose of gp E2. Haematological findings showed that the vaccines did not cause leucopenia or lymphocytopenia and the number of neutrophils and eosinophils during the observation period was within physiological range. The results of the proportion of CD4a+, CD5a+, CD8a+, wCD21+, CD45RA+, CD45RC+ , non-T non-B, SWC3a+ and CD11b+ cells were gained by single-colour flow cytometry. At the end of the trial a significantly increase of percentage of CD4+, CD5a+, CD8+, wCD21+ cells has been found in pigs that received the subunit vaccine and the percentage of CD4+, CD5a+, CD8+, CD45RA+ and CD45RC+ cells was higher in pigs that received the attenuated vaccine. Twenty-eight days after vaccination the percentage of CD4+, CD45RA+ and CD45RC+ was significantly higher in pigs vaccinated with the C strain than in pigs vaccinated with the subunit vaccine. In contrary, the percentage of the wCD21- cells was higher in pigs that received the subunit vaccine. Statistically higher values of SWC3a+ and lower values of CD11b+ cells was observed in pigs that received the attenuated vaccine than in pigs vaccinated with the subunit vaccine. Taken altogether, our results showed that the subunit vaccine produced a better stimulation of B cells and CD11b+ monocytes/macrophages /granulocytes/NK cells, whereas the attenuated vaccine induced a higher response of Th cells, naive/memory cells and macrophages/neutrophils. Thus, both vaccines were able to influence the porcine immune system, by activating different subsets of the immune effector/accessory cells.     

鸭疫里默氏菌病油佐剂灭活疫苗的研制

为了有效控制鸭疫里默氏菌病的发生,我们将本地区分离鉴定的鸭疫里默氏菌作为菌种,采用液体增菌培养后灭活,加入佐剂制成油乳剂灭活疫苗,经无菌和安全检验合格后免疫4日龄雏鸭(0.5mL/只)。结果表明该疫苗免疫效果良好,一次免疫后保护率达70%,两次免疫后保护率达95%。     

鸭疫里氏杆菌-大肠杆菌二联灭活疫苗的研制

从河北某肉鸭场的发病鸭中分离出鸭疫里氏杆菌（Riemerella.anatipestifer，RA）BJ-D-1株，对该菌株采用液体培养、固体培养及鸡坯培养等三种不同方法进行增菌培养，然后与大肠杆菌O1、O2、O78株及河北地方大肠杆菌流行株等的浓缩菌液以一定比例混合，制成油乳剂灭活苗。皮下免疫7日龄雏鸭0.5mL/只。在河北某发病肉鸭场及周围肉鸭场进行田间试验，效果良好。     

自家苗治疗连城白鸭浆膜炎的效果研究

选择刚死亡且具有典型鸭传染性浆膜炎病变的病死鸭,无菌采集其肝组织,对疑似鸭疫里默氏杆菌进行分离鉴定,证明确为鸭疫里默氏杆菌,然后将该菌株扩增培养,通过甲醛灭活,再经无菌试验和安全性试验验证,制成安全的自家苗,随后对自家苗作临床效力试验,结果证明该苗具有较好的免疫防治效果,适合在当地推广使用。     

鸭疫里默氏杆菌多价灭活苗研制的研究报告

用三个血清型的菌株,分别是JYL-1、JYL-2、JYL-7,用两种佐剂,分别是蜂胶和油乳剂共制备了两种多价菌苗,即JYL-1、JYL-2、JYL-7（1、2、7三个血清型）三个RA血清型混合多价蜂胶苗和油乳剂苗。试验结果表明。蜂胶复合佐剂多价苗具有产生免疫保护速度快,免疫持续时间长的优点,接种后第3天即产生部分免疫保护力,第120天时仍具有完全保护力;油乳剂多价苗产生保护力的速度较慢,接种后第10天时开始表现出部分免疫保护,其完全保护力也可持续到接种后第120天。免疫后13d时,免疫保护率可达90％左右。免疫后16d时保护率为100％。免疫后120d时,两种多价菌苗的免疫保护率均可达到100％,免疫后150～180d时,免疫保护率可达800左右。统计学分析证明,菌苗的安全性良好。用3个免疫剂量的菌苗所做的安全试验表明无明显不良反应,菌苗在4℃保存一年,18～22℃室温保存4个月不影响菌苗的免疫效力。田间试验表明,用菌苗免疫鸭群后,可有效地使鸭群抵抗鸭疫里默氏杆菌的感染。     

一株鸭疫里默氏杆菌自然弱毒株的分离鉴定

从四川德阳某鸭场病死鸭肝脏中分离的DY9株细菌,经理化特性、16SrRNA核苷酸序列与cam基因检测,证明为鸭疫里默氏杆菌,并对其毒力与毒力稳定性等进行了测定。DY9株细菌对14日龄非免疫健康鸭的LD50大于2.3×1010CFU/0.5mL,但死亡鸭不具有纤维素性炎症病变,并未能回收到细菌;经敏感鸭体内连续传10代和经巧克力营养琼脂平板传30代后毒力未见显著改变;培养滤液不影响鸭胚成纤维细胞的形态;油佐剂灭活菌苗与活菌苗免疫鸭后14d攻毒分别获得100%和79.52%的相对保护率,21d时均为100%。以上结果表明,DY9株细菌为毒力稳定的鸭疫里默氏杆菌自然弱毒株。     

Leukocyte subsets and specific antibodies in pigs vaccinated with a classical swine fever subunit (E2) vaccine and the attenuated ORF virus strain D1701

Total white blood cell (WBC) counts and percentages of CD4a+, CD8a+, CD5a+, CD45RA+, CD45RC+, wCD21+ and SWC3a+ cells in the peripheral blood of pigs were analysed in this study. Blood samples were collected before and on days 4, 10, 21 and 28 after vaccination. Group 1 pigs were vaccinated with a subunit E2 vaccine (gp E2 32 microg/dose), and Group 2 received a subunit vaccine combined with an attenuated ORF virus strain D1701 10(6.45) TCID50/dose. Control pigs received a placebo. The total WBC count and percentage of particular cell types were within the normal range in vaccinated and control pigs. Although the mechanism of attenuated ORF virus activity is not clear, changes were observed in CD4a+, CD5a+, CD8a+, CD45RA+ and CD45RC+ cells in pigs that received the combination of a subunit vaccine and ORF virus. However, the percentage of wCD21+ and SWC3a+ did not differ significantly from that recorded in pigs given only the subunit vaccine. At days 4 and 10 the number of pigs positive to E2 antibodies was higher in the group that received the subunit vaccine and ORF virus than in pigs vaccinated with the subunit vaccine only. A higher percentage of memory cells (CD45RC+) as well as Th and Tc lymphocytes in pigs that received the ORF virus and the subunit vaccine could be ascribed to a nonspecific influence of the ORF virus on the development (through cognate interactions between T and B cells) and the duration (presumed according to the finding of the clonal expression of memory cells) of humoral immunity (assessed by a higher number of seropositive pigs in this group). This seems likely since the proportion of these cells was found to be lower in the pigs that received E2 vaccine only.     

Compensatory Growth and its Effect on Muscularity and Technological Meat Quality in Growing Pigs

This study investigated the effect of various feeding levels from weaning (day 28) to day 170 of age on growth, muscularity and technological meat quality in female pigs. From day 28 to day 90 of age (growing period) and from day 90 to day 170 of age (finishing period), the pigs were fed either ad libitum (A) or restrictively (R) in a 2 2 2 factorial design with treatments named AA, AR, RA and RR. In the growing period, the growth rate of A pigs was 35% higher than that of R pigs. In the finishing period, the growth rate was dependent on the feed intake in the growing period, i.e. pigs fed restrictively in the growing period had 6-8% higher growth rate in the finishing period (RA and RR) than pigs fed ad libitum in the growing period (AA and AR). Furthermore, despite RA pigs being 11 kg lighter at day 90 of age they produced as much muscle tissue at slaughter as did AA pigs, but less subcutaneous fat, which resulted in a 5% higher meat content of the carcass. The increased muscle growth of RA pigs in the finishing period (compensatory growth) was probably accomplished by increased satellite cell proliferation (muscle DNA accumulation) and increased capacity for protein synthesis, as indicated by a higher RNA concentration. Feeding level did not affect the lightness of meat, the ultimate pH or the drip loss. However, a change in feeding level at day 90 of age (RA and AR) led to a reduction in meat redness. The present data suggest that feed restriction in the growing period results in compensatory growth of muscle tissue in the finishing period if ad libitum feeding was applied during this period, accomplished by increased satellite cell proliferation and increased capacity for protein synthesis, without significantly affecting the technological meat quality.