鸡巨型艾美耳球虫卵囊呼吸作用的研究

采用纯种巨型艾美耳球虫(Eimeria maxima)卵囊,人工感染3周龄的石歧杂雏鸡,及时收集卵囊,以0.1N硫酸溶液悬浮,用微量呼吸检压仪检测卵囊呼吸作用。试验表明:孢子化过程中最初十小时内该球虫卵囊的耗氧量为3.59μl/小时/10~6个卵囊。耗氧量的变化与其细胞分裂过程相对应,耗氧量在三次核分裂前夕降低,但在分裂期升高。最高耗氧量出现于子孢子形成期,达18.91μl/小时/10~6卵囊。孢子化后卵囊的呼吸作用较缓慢,耗氧量为0.85μl/小时/10~6个卵囊。完成孢子化所需时间为26小时。     

柔嫩艾美耳球虫毒害艾美耳球虫变位艾美耳球虫交叉免疫试验

柔嫩艾美耳球虫毒害艾美耳球虫变位艾美耳球虫交叉免疫试验黄兵，赵其平，何林华，吴薛忠，陈兆国，史天卫（中国农科院上海家畜寄生虫病研究所，２００２３２）鸡感染球虫后会产生一种抵抗再次感染的免疫反应，这种免疫反应具有种的特异性，即感染某种球虫不能保护抵抗另...     

堆型艾美耳球虫与柔嫩艾美耳球虫卵囊最短孢子发育时间的测定

将收集到的新排出的堆型艾美耳球虫(E.acervulina)与柔嫩艾美耳球虫(E．tenella)卵囊于29℃静态系统中孢化。以30min为间隔，对卵囊孢子发育的4个阶段在显微镜下进行观察。以1h为间隔，用发育中的卵囊连续接种无球虫鸡，以确定最短孢子发育时间(MST)。对影响孢子发育的外部因素进行了初步探讨。用堆型艾美耳球虫比较不同卵囊浓度，不同悬液深度在卵囊孢化过程中第20h时子孢子形成的卵囊百分率。E.acervulina与E.tenella的MST分别为11b与18b。卵囊浓度和悬液深度与孢子发育时间呈明显反比关系。     

柔嫩艾美耳球虫卵囊DEAE凝胶纯化法的建立

目的建立柔嫩艾美耳球虫卵囊DEAE凝胶纯化方法。方法常规方法从鸡盲肠内容物或粪便漂浮、离心获取卵囊,孢子化后经初步纯化,用EDAE凝胶阴离子交换再次纯化,同时进行了2种缓冲液的选择。结果缓冲液1效果更好,回收率达到79．2％,得到纯度更高。结论本研究建立了的柔嫩艾美耳球虫卵囊纯化方法简单易行,纯化效果好。     

斯氏艾美耳球虫子孢子的超微结构

斯氏艾美耳球虫子孢子呈香蕉形,外被三层膜组成的表膜,有前后极环;顶复合器具典型结构,棒状体为８根;核一个,位于虫体前半部,含一个偏心核仁;相当于核中部水平位置的表膜上,见有一微孔;折光体在核后,呈球形,约占子孢子纵长的２／５;尾部常见一兔尾巴式的结构,构成方式类似阿米巴的伪足,是由被表膜的胞质向外突出形成的;胞质中见有许多管泡状线粒体和一些电子致密体。     

斯氏艾美耳球虫子孢虫的超微结构

斯氏艾美耳球虫子孢子呈香蕉形,外被三层膜组成的表膜,有前后极环;顶复合器具典型结构,棒状体为８根;核一个,位于虫体前半部,含一个偏心核仁;相当于核中部水平位置的表膜上,见有一微孔;折光体在核后,呈球形,约占子孢子纵长的２／５;尾部常见一兔尾巴式的结构,构成方式类似阿米巴的伪足,是由被表膜的胞质向外突出形成的;胞质中见有许多管泡状线粒体和一些电子致密体。     

毒害艾美耳球虫卵囊壁蛋白的SDS-PAGE及Western blot分析

利用聚丙烯酰胺凝胶(SDS-PAGE)电泳技术,分析了毒害艾美耳球虫未孢子化卵囊壁可溶性蛋白。用鼠抗毒害艾美耳球虫重组配子体蛋白rEnGAM56和rEnGAM59多克隆抗体,对卵囊壁可溶性蛋白进行了Western blot分析。结果显示,至少有24条较清晰的电泳条带,其中6条为相对明显的主带,其相对分子质量分别为37 000,35 000,34 000,20 000,14 000和12 000。Western blot分析显示,鼠抗rEnGAM59和rEnGAM56多抗均能识别1条条带,前者的相对分子质量约14 000,后者约30 000。推测30 000和14 000卵囊壁蛋白分别来自毒害艾美耳球虫配子体蛋白EnGAM56和EnGAM59。     

斯氏艾美耳球虫致病性研究

为了深入研究斯氏艾美耳球虫的致病性,通过给幼兔人工感染不同数量的斯氏艾美耳球虫孢子化卵囊,对其发病后的急性临床症状及病理变化进行观察与分析。将幼兔分为4组,空白对照组和3个试验组。除空白对照组外,3个试验组家兔分别经口感染斯氏艾美耳球虫孢子化卵囊2×105个/只、1×105个/只、0.5×105个/只。结果表明,在感染后第8天,3个试验组和空白对照组家兔的每克粪便卵囊数(OPG)分别为2.19×107、2×107、1×107和0,相对增重率分别为38.9%、58.0%、73.3%和100%。表明斯氏艾美耳球虫对幼兔有很强的致病性。     

11株巨型艾美耳球虫繁殖力的比较研究

用巨型艾美耳球虫扬州株、南通株、连云港株、苏州株、凤阳株、青岛株、福州株、龙岩株、广州株、上海株和美国株孢子化卵囊100个/只或10 000 个/只,经嗉囔接种7 日龄或21 日龄无球虫黄羽肉公雏,以麦克马斯特法对感染后第6 天至第14 天间每天24 h排出的卵囊计数并进行统计分析。结果表明,各虫株的排卵囊量有差异,100个/只卵囊感染7日龄雏鸡时,广州株和凤阳株排卵囊量最多,上海株和苏州株排卵囊量最少;100个/只卵囊感染21 日龄雏鸡时,扬州株排卵囊量最多,上海株排卵囊量最少;10 000 个/只卵囊感染21 日龄雏鸡时,苏州株排卵囊量最多,连云港株和扬州株的排卵囊量最少;排卵囊量随着感染剂量的增加或鸡日龄的增大而增多;在同一感染剂量时,福州株和连云港株增加最多,上海株和苏州株增加最少;在同一感染鸡日龄时,苏州株和上海株增加最多,南通株和扬州株增加最少;各虫株的排卵囊规律极相似,在感染后第7 天排卵囊量达峰值,第8 天、第9 天排出的卵囊量显著减少,第10天排出的卵囊量仅占总量的2%,第14天接近于0%;第6天至第8天排出的卵囊量占总量的87%~99%。     

斯氏艾美耳球虫病的研究进展

斯氏艾美耳球虫（E．stiedai）是寄生在肝脏胆管上皮细胞内的一种危害较大、致病性较强的兔球虫,最初由Leeuwenhoek（1674年）等发现,但是直到1859年才由Lindermannt将其正式命名为球虫。斯氏艾美耳球虫病在国内平均感染率为22％,死亡率80％左右,给我国养兔业造成巨大的经济损失。     

Epidemiological Investigation and Species Identification of Eimeria of a Rex Rabbit Farm in Fujian Province

In order to investigate coccidium infection of a Rex rabbit farm in Fujian province, 494 fecal samples were collected from different ages of Rex rabbits and examined by McMaster's method and the method of saturated saline flotation.The results showed that the total prevalence was 87.45% and the mean oocysts per gram (OPG) value was 2.56×10⁴.The prevalences of coccidia among each rabbit age were different, and it was highest at the age of about 40 d (98.61%) and about 65 d (98.95%) rabbits.The prevalence and OPG values at the age under 30 d (19.23% and 331) were significantly lower, and the distribution pattern of OPG values was negative binomial distribution in rabbits by using fitting of distributions.In total, 11 species of Eimeria were identified from oocyst-positive samples.The infection of Eimeria was mixed infection with more than two Eimeria species, and mostly of which were infected by 3 to 7 Eimeria species.The dominant species of Eimeria were E.perforans, E.media, E.exigua and E.magna, and their prevalences were 20.15%, 16.79%, 13.94% and 13.48%, respectively.     

福建某獭兔场兔艾美耳球虫感染情况调查及种类鉴定

为掌握福建某规模化獭兔场兔球虫病的流行情况,采用麦克马斯特计数法(McMaster's method)和饱和氯化钠溶液漂浮法,对采集的494份獭兔粪便样品中兔球虫感染情况和卵囊种类进行调查分析。结果显示,该獭兔场兔球虫总感染率和平均每克粪便卵囊数(OPG值)分别为87.45%和2.56×10⁴,各日龄段均有感染,其中以40日龄左右兔和65日龄左右兔感染率最高,分别达98.61%和98.95%;30日龄以下哺乳仔兔感染率和OPG值均最低,分别为19.23%和331,且该场兔球虫卵囊的OPG值分布符合负二项分布。本次调查共检出11种兔艾美耳球虫,均为混合感染,感染球虫种类多为3~7种;其中感染比例最高的4种兔球虫依次为穿孔艾美耳球虫(20.15%)、中型艾美耳球虫(16.79%)、小型艾美耳球虫(13.94%)和大型艾美耳球虫(13.48%)。     

Sequence and Phylogenetic Analysis of rDNA ITS of Three Eimeria Species in Rabbit

In order to study whether the internal transcribed spacers (ITS) sequence could be used as a molecular marker for the species identification of rabbit coccidian, the rDNA ITS of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were amplified by polymerase chain reaction (PCR), and were cloned into pGEM-T Easy vector subsequently. The positive recombinant plasmids were identified by PCR and then sequenced. By sequence comparison and comparative analysis with the relative sequences of rabbit Eimeria spp. available in GenBank, the results showed that the lengths of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were 1065, 1009 and 1047 bp, respectively, and the sequence homologies with the same species sequences were 99.2%, 99.0% and 94.5%, respectively, while were 55.3% to 82.1% compared with corresponding sequences of other different species sequences. The phylogenetic analysis using software Mega 5.0 showed that all rabbit coccidia clustered together in a clade, which was divided into two sister lineages, corresponding to the presence or absence of oocyst residuum. The result demonstrated ITS could be used as a molecular marker for the species identification of rabbit coccidia.     

基于ITS-1序列的5种兔球虫系统进化分析

利用核糖体DNA内转录间隔区1(ITS-1)序列对兔艾美耳球虫进行系统进化分析,并探讨生物学和形态学特征在兔球虫进化中的意义。单卵囊分离大型、黄、肠、中型及穿孔艾美耳球虫卵囊,接种无球虫兔获得纯种卵囊,CTAB法提取卵囊基因组DNA,PCR扩增ITS-1区后克隆、测序。将测序结果与GenBank发布的兔球虫ITS-1序列进行比对和遗传距离分析,绘制系统进化树。结果显示:大型、黄、肠、中型及穿孔艾美耳球虫河北株ITS-1序列分别长320、330、351、336和341bp。5种兔球虫河北株与GenBank中同种兔球虫ITS-1序列相似性分别为96.9%、97.3%、96.9%、99.1%和99.4%。兔球虫形成单系群,该单系群分为2个姐妹谱,与卵囊残体有无相对应,其它形态学和生物学特征与系统进化无相关性。研究结果表明外残体的有无可作为兔球虫进化分类的特征。     

齐卡肉兔球虫种类调查研究

对西宁市某种兔场进行了球虫种类调查，发现兔艾美尔球虫8种：穿孔艾美尔球虫、大型艾美尔球虫、中型艾美尔球虫、小型艾美尔球虫、无残艾美尔球虫、盲肠艾美尔球虫、斯氏艾美尔球虫和梨形艾美尔球虫，其中以穿孔艾美尔球虫所占比例最高，为55.5%，其次为大型艾美尔球虫，占17.5%，中型艾美尔球虫占11.5%，盲肠艾美尔球虫占8%。观察、研究了卵囊的形态、大小、颜色、内部构造及孢子化时间等。     

柔嫩艾美耳球虫大配子发育和卵囊的超微结构

利用透射电镜对柔嫩艾美耳球虫大配子发育和卵囊的超微结构进行了观察。结果表明，大配子体和小配子体寄生于相邻的宿主细胞内，相伴产生。大配子体的发育、大配子的形成、受精及卵囊形成均于带虫空泡内完成。大配子体系由末代裂殖子侵入肠上皮细胞后，缩短、长大、变圆而形成，大配子体和大配子外被单位膜，细胞核位于大配子体中央，早期大配子体的细胞核有1个明显的核仁，细胞质内逐渐形成大量成囊体1、成囊体2、大量支链淀粉和脂肪体，成囊体1比成囊体2形成的早。卵囊壁有5'层，内有大量支链淀粉和指肪体。含有大配子体、大配子和合子的带虫空泡内有丰富的泡内小管，它们是大配子体、大配子和合子获得营养物质和排出代谢废物的途径。     

5株柔嫩艾美耳球虫对4种抗球虫药的抗药性

选择260只19日龄雏鸡，随机分成26组，每组10只，研究柔嫩艾美耳球虫(E．tenella)南京株、凤阳株、扬州株、宣城株和蒙城株对地克珠利(Diclazuril)、马杜霉素(Maduramicin)、氯羟吡啶(C1opido1)、氯苯胍(Robenidine)的抗药性。每株球虫均设4个感染用药组、1个感染不用药组，另外设1个不感染不用药组。以POAA、RLS、ROP、ACI作为指标，判定5株球虫对4种药物的抗药性。结果表明凤阳株、扬州株和蒙城株分别仅对马杜霉素、氯苯胍、地克珠利敏感，南京株和宣城株对该4种药物均已产生不同程度的抗药性。提示目前在凤阳、扬州和蒙城地区可分别合理地使用马杜霉素、氯苯胍和地克珠利，对其它药物须减少或暂停使用。     

Eimeria acervulina Infections from Each of the Four Sporocysts of a Single Oocyst

Oocysts were found in all four birds each infected with one of the four sporocysts of a single oocyst of Eimeria acervulina and in three of the four birds infected with single sporocysts from two other oocysts. Possible use of these infections for genetic studies was discussed. An efficient method for collecting fecal material containing sometimes rather low levels of oocysts was also described.     

Comparison of Production Performance of Different Mating Combination Between Castor Rex Rabbit and Plicae Rex Rabbit

In order to compare the production performance of different mating combination between Castor Rex rabbit and Plicae Rex rabbit,40 Castor Rex rabbit (10 ♂ and 30 ♀) and 40 Plicae Rex rabbit (10 ♂ and 30 ♀) with 6 to 7 month old were selected.Hybrid testing was conducted using Plicate Rex rabbit and Castor Rex rabbit as crossing parents which were Castor Rex rabbit pure breeding (Castor Rex rabbit ♂×Castor Rex rabbit ♀),Plicate Rex rabbit pure breeding (Plicae Rex rabbit ♂×Plicae Rex rabbit ♀),orthogonal combination (Castor Rex rabbit ♂×Plicae Rex rabbit ♀)and reciprocal combination (Plicae Rex rabbit ♂×Castor Rex rabbit ♀).The reproductive performance of rabbits does and the growth performance,the slaughter performance and wool quality of their offsprings were measured respectively. The results showed that the neonatal weight of litter,litter size,lactescence,survival rate,weaning weight of litter,F/G,slaughter rate,skin area,hair density,hair fineness and coarse wool rate of orthogonal combination were 320.71 g,6.71,1 727.43 g,83.91%,5 304.14 g,4.06,62.43%,1 160.20 cm²,15 951.70/cm²,16.32 μm and 6.83%,respectively,while that of reciprocal combination were 315.14 g,6.52,1 710.00 g,80.19%,4 814.82 g,4.30,62.93%,1 172.20 cm²,16 092.01/cm²,16.43 μm and 6.78%,respectively,and the differences between reciprocal combination and orthogonal combination were not significant (P>0.05).The F/G,slaughter rate and coarse wool rate of F1 in Castor Rex rabbit pure breeding were higher than that of Plicate Rex rabbit pure breeding,while the other indexes were all lower than Plicate Rex rabbit pure breeding,of which the differences between two pure breeding, except F/G,slaughter rate,hair fineness and coarse wool rate, were significantly different (P<0.05).In conclusion,the reproductive performance of parental generation and growth and slaughter performance of F1 in orthogonal combination was slightly better than that of reciprocal combination,while the wool quality of F1 in reciprocal combination was better than orthogonal combination.