毒害艾美球虫二次感染雏鸡外周血液免疫功能的变化

将180只14日龄雏鸡随机分为毒害艾美球虫（Eimeria necatrix）初次感染组、二次感染组和对照组，应用免疫SPA菌体花环、间接ELISA及细胞培养技术和四甲基偶氮唑盐（MTT）测定法对相关免疫学指标进行了检测，以研究毒害艾美球虫二次感染对雏鸡外周血液免疫功能变化的影响。结果显示，毒害艾美球虫二次感染雏鸡外周血液T、B淋巴细胞数量及其对ConA或PMA的增殖反应和血清IgG、IgM、IgA免疫球蛋白含量均不同程度地高于未感染毒害艾美球虫的对照组雏鸡。证实毒害艾美球虫二次感染雏鸡外周血液的细胞免疫和体液免疫功能均明显提高。     

毒害艾美耳球虫纯种卵囊收集鉴定及致病性测定

鸡球虫是养鸡业的一大疾病 ,严重阻碍着我国养鸡业的发展。在各种鸡病中 ,球虫病的发生率最高 ,占 1 /6～ 1 /5。Rose和 Long( 1 980 )报道英国养禽业因球虫病每年损失 5 5 0万英镑 ,其中一半是药物费用 ;在美国损失可能达到 5 0 0 0万英镑。 Rose和Long( 1 985 )保守地估计全世界养禽业每年球虫造成的损失为 5亿美元左右 ,其中 2 .5亿美元用于药费支出 [1]。中国是养鸡大国 ,养鸡数量居世界之首 ,但由于集约化程度低 ,饲养环境差 ,保健系统不完善因素的影响 ,球虫病所造成的损失 ,可能高于世界平均水平。球虫病所造成的经济损失至少 5 0 …     

Immunohistochemical study of S100-like protein in Eimeria brunetti and Eimeria acervulina

We have investigated the expression of a calcium-binding protein, the S100 protein, in Eimeria brunetti and Eimeria acervulina stages. For this purpose, paraffin sections of distal ileum and bursa of Fabricius or duodenum from experimentally infected chickens were treated with anti-alpha-S100 (anti-alpha subunit of S100 protein) and anti-beta-S100 (anti-beta subunit of S100 protein) monoclonal antibodies and anti-S100 whole molecule polyclonal antibody. The avidin-biotin peroxidase method was used to demonstrate immunoreactivity. In the ileum, our results reveal a positive immunoreaction for the beta subunit and S100 whole molecule within the macrogametes of E. brunetti, whereas they were devoid of immunostaining after treatment of the paraffin sections with the anti-alpha-S100 antiserum. Schizonts and oocysts of E. brunetti and all the E. acervulina stages gave a negative reaction after treatment with any of the three antiserum used in the study. This result indicated that the S100 protein molecules within these stages were not recognized by the antibodies, suggesting that these molecules are different from those identified in macrogametes of E. brunetti. By contrast, in the epithelial cells, lining the lumen of the bursa of Fabricius, macrogametes of E. brunetti were stained by the three antibodies used. These results may indicate the existence of metabolic adaptations that enable the parasite to invade tissue sites different from those where the parasite usually develops.     

Effects of Eimeria brunetti infection and dietary zinc on experimental induction of necrotic enteritis in broiler chickens.

Broilers infected with Eimeria brunetti and given dietary zinc were examined for experimental induction of necrotic enteritis. Inoculation with sporulated E. brunetti oocysts at 7 days of age was followed by 5 consecutive days of oral inoculation with cultured Clostridium perfringens. Feed was supplemented with zinc at 1000 ppm. Upon necropsy of broilers 6 days after coccidial inoculation, necrotic enteritis was found in 20% (2/10) of birds given both organisms and dietary zinc. Coccidial lesion scores were also highest in that group. Birds infected with E. brunetti and C. perfringens with no dietary zinc had significantly higher coccidiosis lesion scores (P less than 0.05) than groups inoculated with E. brunetti only, regardless of zinc supplementation. Alpha toxin levels in intestinal contents were low in groups infected with both organisms, regardless of zinc supplementation. Zinc was tested for effects of alpha toxin production in vitro. In the mid-log phase (6 hours incubation), a high level of alpha toxin was produced in zinc-supplemented media, but this was lost quickly in the presence of trypsin. Addition of zinc partly protected the toxin from the action of trypsin.     

布氏艾美耳球虫（Eimeria brunetti）早熟虫株的免疫原性及毒力

用５００卵囊／只剂量的Ｅｂｒｕｎｅｔｉ早熟虫株经口服免疫３日龄公鸡雏后１４ｄ,再以４５×１０４卵囊／只剂量的亲本毒株口服攻毒。结果,免疫雏鸡受到完全保护,其平均增重与空白对照组差异不显著。当免疫剂量为２５０卵囊／只时,仅对攻毒具有部分保护作用。毒力试验结果表明,Ｅｂｒｕｎｅｔｉ早熟虫株的致病力显著低于Ｅｂｒｕｎｅｔｉ强毒虫株。当感染剂量相同时,早熟虫株感染组的平均增重与强毒虫株感染组差异显著或极显著。由此认为,Ｅｂｒｕｎｅｔｉ早熟虫株是一弱毒虫株,可用于鸡球虫弱毒疫苗的制备。     

毒害艾美耳球虫-田间分离株的致病性试验

毒害艾美耳球虫扬州田间分离株以不同的剂量(10 000,30 000,50 000,70 000,90 000个卵囊/羽)感染28日龄黄羽肉雏鸡,以平均增重、死亡率、病变记分等指标评价其致病性。结果显示,除10 000个卵囊组外,其余4个剂量组均显著影响增重(P<0.05),并引起鸡的死亡,死亡率分别为37.5%～50%、37.5%～50%、62.5%、75%～87.5%;各感染组均出现肠道病变,其中10 000个卵囊组的病变值显著小于其余4个剂量组(P<0.05),但与不感染对照组相比有显著差异(P<0.05)。结论表明,该毒害艾美耳球虫田间分离株有很强的致病性。     

鸡球虫病四价活疫苗对毒害艾美耳球虫和柔嫩艾美耳球虫的免疫效果观察

为评价一种商品化鸡球虫病四价活疫苗[柔嫩艾美耳球虫(E.tenella)、毒害艾美耳球虫(E.necatrix)、巨型艾美耳球虫和堆型艾美耳球虫]对E.necatrix和E.tenella的免疫保护效果,本研究基于试验鸡血便记分、存活率、相对增重率、病变值、卵囊值及抗球虫指数(ACI)等指标对其免疫效果进行检测.结果表明,免疫攻虫组抗E.necatrix和E.tenella的ACI分别为157.3和135.6,均低于160,表明该球虫疫苗抗两种球虫的效果均为低效.然而,免疫攻虫组鸡血便数量、血便记分、增重、卵囊产量和病变记分等指标均优于阳性对照组.综合指标显示,该疫苗对E.necatrix和E.tenella具有一定的免疫保护力,但单纯采用疫苗并不能够达到防控鸡球虫病的目的.     

鸡布氏艾美耳球虫致病性研究

用１８日龄公雏作为试验对象,以增重、病变和死亡为判定指标,用Ｅ．ｂｒｕｎｅｔｉ的０．５万／只、１万、５万、１０万、２０万、５０万和１００万／只为接种梯度,研究了布氏艾美耳球虫的致病性。结果表明：接种０．５万／只至２０万／只剂量的Ｅ．ｂｒｕｎｅｔｉ都明显影响鸡的增重;（与对照组相比,均存在极显著性差异（Ｐ＜０．０１）,各剂量组间增重无差异,）５０万／只和１００万／只剂量组的鸡出现减重。５０万／只剂量组鸡死亡５０％,１００万／只剂量组鸡死亡１００％;Ｅ．ｂｒｕｎｅｔｉ接种５万／只剂量组及以上各组鸡饮食欲消失,鸡失去自洁行为,粪量少,含有粘液和血液;剖检病变主要在小肠至直肠,肠壁变薄,易碎,呈粉红色至暗红色,肠粘膜上有小的出血点,肠内容物以粘液和血液为主。部分鸡的盲肠内容物干、呈暗红色。（病变计分为：对照组０分,０．５万／只为０．５７＋;１万／只０．０３＋;５万／只１．１７＋;１０万／只１．５０＋;２０万／只２．２３＋;５０万／只３．６４＋;１００万／只３．８３＋。）１００万／只剂量组中有两只鸡出现腿麻痹症状,不能站立。     

布氏艾美耳球虫保定株的鉴定及致病性研究

本研究采用琼脂薄膜球虫单卵囊分离技术,从河北省保定市暴发球虫病的某鸡场鸡粪便中成功分离出布氏艾美耳球虫卵囊。生物学特性研究结果表明,该虫种潜隐期为125 h,卵囊大小为26.1 μm×21.3 μm,形状指数为1.22,寄生与病变部位主要在小肠后段、直肠和泄殖腔,特征性病变是肠黏膜层有出血斑纹及血性内容物。感染一定剂量该球虫,鸡的生产性能受到影响,当感染剂量为2×10⁵个/羽时会出现鸡死亡,表明该虫株具有较强致病性;应用特异PCR方法对获得的卵囊进行分子鉴定,结果确认所分离和纯化的球虫卵囊为布氏艾美耳球虫纯种,并将该虫株命名为布氏艾美耳球虫保定株--BBD株。     

毒害艾美耳球虫晚熟系生物学特性研究

本文对毒害艾美耳球虫晚熟系基本生物学特性进行了研究。结果表明，晚熟系的致病性与同等剂量中熟系的相当；繁殖能力高于中熟系，且每羽鸡的排卵囊总量与剂量成正相关，每个卵囊的繁殖能力与剂量成负相关；在同等剂量免疫下，晚熟系的免疫原性好于中熟系；2次免疫后，晚熟系免疫组的保护力与中熟系的相当，攻虫后卵囊繁殖减少率均在99％以上。     

Pleurotus ostreatus extract inhibits Eimeria species development in naturally infected broiler chickens

Tropical Animal Health and Production - Coccidian organisms of poultry have proved very hard to control due to their abilities to resist most anticoccidial drugs. Pleurotus ostreatus (Fr.) Jacq. ex...     

毒害艾美耳球虫配子体gam22基因的克隆与序列分析

提取毒害艾美耳球虫(En)配子体总RNA,应用RT-PCR进行gam22基因的克隆,测序后对其进行序列分析。结果表明,Engam22基因全长为713 bp,其中包含一个完整的开放阅读框,大小为561 bp,编码186个氨基酸,含有一个富含组氨酸和脯氨酸的特征性结构域;与柔嫩艾美耳球虫(Et)配子体Etgam22基因序列进行比对,同源性为97.7%。此配子体蛋白基因具有较高的保守性,可以作为抗球虫疫苗的候选基因。     

Molecular identification of Eimeria species infecting market-age meat chickens in commercial flocks in Ontario

A previously described multiplex PCR was evaluated for the identification and prevalence of Eimeria species in market-age commercial chicken flocks in Ontario. The multiplex PCR based on species-specific RAPD-SCAR markers was able to distinguish six available laboratory strains of Eimeria species (E. tenella, E. maxima, E. necatrix, E. mitis, E. acervulina, and E. brunetti) and E. tenella, E. maxima and E. acervulina in unknown field samples, including multiple infections in single reactions. No backyard (0/77) and 20/360 market-age commercial chickens were oocyst-positive using standard fecal flotation methods. PCR identified E. tenella alone (9/360, 2.5%), E. maxima alone (5/360, 1.38%), E. maxima plus E. tenella (5/360, 1.38%) and E. acervulina alone (1/360, 0.27%) in market-age commercial broilers. This is probably the first time the multiplex PCR has been evaluated in poultry establishments in Canada and illustrates the value of the tool in coccidiosis epidemiology on commercial farms.