鸡巨型艾美耳球虫单卵囊分离及雏鸡扩增试验

采用饱和食盐水漂浮法收集就诊病死球虫阳性鸡小肠中段内容物中的球虫卵囊,恒温培养至孢子化后,用2%琼脂薄板进行球虫单卵囊分离,分别感染7只5日龄雏鸡,对据形态学鉴定为巨型艾美耳球虫的分离株进行2代单卵囊分离和雏鸡感染,取其后代以每只1.0×10⁴个卵囊感染10只10日龄雏鸡进行卵囊扩增,获得大量纯种巨型艾美耳球虫卵囊。结果表明,刀片切割琼脂薄板单卵囊分离法操作简便,单卵囊感染成功率较高,准确率达100%,适用于纯种卵囊的分离与扩增。     

鸡柔嫩艾美耳球虫广西株的分离与鉴定

目的建立一种简单、实用的单卵囊分离方法，并对一株广西柔嫩艾美耳球虫进行分离。方法采用电泳制胶槽来制作琼脂块进行球虫单卵囊的分离，单卵囊实验感染9只1日龄雏鸡，感染后收集粪便，用饱和盐水漂浮法进行卵囊检测。纯种卵囊经口感染10只1日龄雏鸡，观测卵囊寄生部位、最短孢子化时间，以及其潜在期和排卵高峰期。结果2只雏鸡粪便中检出卵囊，单卵囊感染成功率为22％。通过对其中一株球虫的研究，根据其卵囊的形状、大小、寄生部位、潜在期、卵囊最短孢子化时间、排卵高峰期等生物特征，鉴定该株球虫为柔嫩艾美耳球虫（Eimeria tenella）。结论本研究成功建立一种单卵囊分离技术，可作为球虫单卵囊分离的常规方法。     

影响鸡球虫单卵囊分离纯化的因素分析

单卵囊分离技术是获得品种单一、基因型纯净的卵囊,以便更有效的鉴别球虫种类和建立球虫纯种体系的重要手段.论文主要从雏鸡日龄、卵囊侵袭力、分离方法和材料的选择三个方面来分析其对鸡球虫单卵囊分离纯化的影响,从而为单卵囊分离技术的改进提供指导,进一步为球虫的各项研究提供基础.     

鸡艾美耳球虫哈尔滨株的分离与鉴定

为研究鸡艾美耳球虫哈尔滨株单一虫种的生物学特性,本研究利用单卵囊分离技术分离了6种鸡艾美耳球虫哈尔滨株,用分离的单卵囊接种雏鸡,同时应用PCR方法对收集的单卵囊分离株进行球虫种类鉴定并进行同源性和系统进化分析.结果显示,分离得到的6株鸡艾美耳球虫分别为柔嫩艾美耳球虫、毒害艾美耳球虫、堆型艾美耳球虫、早熟艾美耳球虫、和缓艾美耳球虫和布氏艾美耳球虫,PCR结果表明单卵囊分离株确为该6种纯株.本研究表明毛细吸管单卵囊分离法简便易行,使接种难度降低,提高了接种成功率,为球虫的分子生物学研究奠定了基础.     

巨型艾美耳球虫合肥(HF)株的分离与鉴定

本试验采用形态学方法分离合肥地区鸡体内巨型艾美耳球虫卵囊,通过雏鸡进行单卵囊感染与扩增试验,用PCR方法扩增该分离株的ITS-1基因序列,并与巨型艾美耳球虫相关序列进行比对、构建系统进化树,从而确定合肥地区巨型艾美     

鸡胚传递配子母细胞疫苗产生的母源免疫力可防止新生雏鸡感染艾美尔球虫

给繁殖母鸡口服接种柔嫩艾美尔球虫鸡胚传递配子母细胞佐剂疫苗,并于加强免疫后2周收集所产鸡蛋进行孵化;出壳雏鸡于7日龄用柔嫩艾美尔球虫(本地分离株)的孢子化卵囊进行攻击,结果免疫母鸡所产雏鸡的球虫卵囊排出量较对照组母鸡所生雏鸡减少70%以上,且无鸡只死亡,而五分之四的对照组雏鸡死于球虫病。这些结果表明,鸡胚传递配子母细胞疫苗所产生的母源免疫力能够防止后代雏鸡受到柔嫩艾美尔球虫的感染。     

海南省文昌鸡主要养殖地区鸡球虫种类及感染情况调查

为了解海南省文昌鸡主要养殖地区鸡球虫种类及感染情况,采用群体采样法分别从各调查点采集新鲜鸡粪,检查粪样。阳性粪样采用饱和盐水漂浮和离心沉淀法进行卵囊分离,2.5%重铬酸钾培养保存。显微镜下观察卵囊进行虫种鉴定,并统计各调查点的鸡球虫种类、感染率和感染强度。结果显示,检查雏鸡(15～42日龄)、青年鸡(43～84日龄)和成年鸡(85～130日龄)粪样各300份,球虫的感染率分别为82.7%、78.0%和38.3%。共检出7种球虫,均属艾美耳属,分别为毒害艾美耳球虫(E.necatrix)、堆型艾美耳球虫(E.acervulina)、巨型艾美耳球虫(E.maxima)、柔嫩艾美耳球虫(E.tenella)、和缓艾美耳球虫(E.mitis)、哈氏艾美耳球虫(E.hagani)和早熟艾美耳球虫(E.praecox)。结果表明,球虫感染在海南省文昌鸡主要养殖地区普遍存在,雏鸡和青年鸡的感染率较高,且中度以上(包括中度)感染偏多;成年鸡的感染率相对较低,且多为轻度感染。     

"肉鸡球虫病及其综合防治措施"专栏(三)鸡胚传递配子母细胞疫苗产生的母源免疫力可防止新生雏鸡感染艾美尔球虫

给繁殖母鸡口服接种柔嫩艾美尔球虫鸡胚传递配子母细胞佐剂疫苗,并于加强免疫后2周收集所产鸡蛋进行孵化;出壳雏鸡于7日龄用柔嫩艾美尔球虫(本地分离株)的孢子化卵囊进行攻击,结果免疫母鸡所产雏鸡的球虫卵囊排出量较对照组母鸡所生雏鸡减少70%以上,且无鸡只死亡,而五分之四的对照组雏鸡死于球虫病.这些结果表明,鸡胚传递配子母细胞疫苗所产生的母源免疫力能够防止后代雏鸡受到柔嫩艾美尔球虫的感染.     

柔嫩艾美尔球虫郑州株的分离与鉴定

鸡球虫病是对养鸡业危害最大的寄生虫病之一。在鸡的多种球虫中,柔嫩艾美尔球虫感染最普遍,致病力最强,致死率最高,危害最严重。为了建立柔嫩艾美尔球虫郑州株纯种体系,为进一步研究该球虫的致病性、免疫防治及临床评价抗球虫药药物效用和球虫耐药性,我们进行了柔嫩艾美尔球虫郑州株的单卵囊分离与感染和单卵囊后代扩大试验,结果如下:     

鸡球虫卵囊分离纯化与单卵囊制备技术的改进

鸡球虫病是严重危害和制约养鸡业发展的寄生虫病之一,凡是有养鸡的地方就有不同程度的鸡球虫病发生,特别是在集约化养鸡场,其发病率可达到50%～70%,死亡率也达20%～30%,严重时高达80%.对鸡球虫卵囊的分离纯化及单卵囊的制备工作是进行球虫病研究分析的主要工作,也是必备的工作.同时,球虫卵囊的分离技术是鉴别球虫的种类和建立球虫的纯种体系、调查某地区内球虫种类和获得球虫的纯种体系的重要手段,对进一步研究球虫的致病性、药物效应、产生的耐药性和禽体对球虫的免疫性、以及对抗球虫药的开发和球虫疫苗研制有重要意义.在结合科研实践的基础上对鸡球虫卵囊的分离纯化及单卵囊制备的操作方法进行改进,改进后提高了虫卵的回收率及单卵囊制备成功率,减化了工作流程.     

Evaluation of interspecific hybrids of the chicken, guinea fowl, and Japanese quail for innate resistance to coccidia

Experimental chicken/guinea fowl hybrids, guinea fowl, and chickens were orally inoculated with Eimeria acervulina or E. tenella, which are specific for chickens, or with E. grenieri, which is specific for guinea fowl. No intact oocysts were found in feces within 24 hr of inoculation, suggesting that excystation occurred in the normal and abnormal hosts. No oocysts were found in the feces of hybrids during a 9-day postinoculation period. The guinea fowl passed oocysts of guinea fowl coccidia (E. grenieri) but not those of chicken coccidia, and the chickens passed oocysts of chicken coccidia (E. acervulina and E. tenella) but not those of guinea fowl coccidia. Some asexual development (schizogony) occurred in hybrids inoculated with E. tenella, but sexual development (gametogony) did not. In contrast, quail/chicken hybrids became infected with oocysts of chicken coccidia (E. acervulina, E. tenella, and E. maxima) and quail coccidia (E. bateri) and passed a few oocysts during the normal patent period; control chickens and quails became heavily infected with oocysts of chicken and quail coccidia, respectively.     

复方球虫散对人工感染鸡球虫病治疗效果的临床试验

为科学、客观地评价复方球虫散对实验性鸡球虫病的防治效果,采用人工接种柔嫩艾美耳球虫孢子化卵囊的方法复制鸡球虫病,使用纯中药组方复方球虫散进行鸡球虫病的临床疗效试验,观察并计算复方球虫散对试验鸡的死亡率、相对增重率、病变值和卵囊值的影响,并求出复方球虫散的抗球虫指数(ACI).试验结果表明:复方球虫散按1.0%拌料混饲,对鸡球虫病有较好的治疗效果.     

湘黄鸡球虫病病原学调查与鉴定

鸡球虫病是危害养鸡业的主要疾病之一,为了解湘黄鸡球虫病病原学流行情况,选择湘黄鸡饲养较集中的衡东、衡山、衡南、衡阳县以及衡阳市郊区五个县区,以地面散养的湘黄鸡为病原学调查对象。通过粪样分肠段采集,采用琼脂薄板单卵囊的分离培养和球虫单卵囊感染试验。经鉴定,结果发现湘黄鸡球虫病的病原体有堆型艾美耳球虫、早熟艾美耳球虫、柔嫩艾美耳球虫、毒害艾美耳球虫、巨型艾美耳球虫等五种球虫。鉴定中,采用4%琼脂糖块将单卵囊分离最合适,此法与其他分离方法比较,简单易行,不需要专门的分离仪器。     

柔嫩艾美耳球虫广西南宁株的分离及鉴定

利用从南宁市郊养鸡场球虫病鸡粪便中收集的球虫混合种卵囊感染小鸡,再应用单卵囊分离感染技术,从感染鸡盲肠中收集的卵囊分离纯化获得1株纯种球虫,经鸡体传代增殖,对该虫株的卵囊大小和卵形指数、潜在期、排卵高峰期、最短孢子化时间、寄生部位、致病性等指标进行观察和测定。结果测得该虫株卵囊的平均大小为(25.743±1.94126)μm×(21.4±1.85985)μm,平均卵型指数为1.2067±0.07;潜在期为140 h;其排卵囊峰期在第6～9天,最高峰在第7天;最短孢子化时间为19 h;寄生部位在盲肠;对两周龄的艾维茵鸡,当使用5×104的孢子化卵囊感染剂量时死亡率为7.5%。根据这些测定和观察到的指标综合鉴定该分离株球虫为柔嫩艾美耳球虫,并命名为柔嫩艾美耳球虫广西南宁株(Eimeria.tenella-GXNN),该研究结果为进一步研究本地区鸡球虫病的药物治疗和免疫预防等奠定了基础。     

Experimental reproduction of necrotic enteritis in the chicken. 1. Mono- and polyinfections with Clostridium perfringens and coccidia with reference to cage managemen]

Experimental mono-infection and poly-infection, using vegetative germs of a CL. perfringens Type A strain 1663 and its toxin as well as E. acervulina, necatrix or mitis oocysts, were applied to 100 SPF chicken aged seven days. While clostridial or coccidial mono-infection did not cause any loss and only unspecific pathologic-anatomic changes, polyinfection was accompanied by diarrhoea and slower weight increase, with 55.4 per cent of the chicken having been lost three to nine days from the outbreak of the infection. The pathologic-anatomic findings recorded from the chicken with poly-infections included haemorrhagic, ulcerative, and necrotic intestinal inflammations, primarily in the small intestin. The results of these experimental infections indicated a possible correlation between CL. perfringens and coccidia in the pathogenesis of necrotic enteritis.     

Internal parasites in calves and heifers in a central rearing barn

In the specialised rearing house for young cattle of all age groups the occurrence of endoparasites was followed. In the age group of 21 to 90 days, 56% of calves aged about 30 days eliminated oocysts of coccidia. On the whole, 49.3% of calves were infested by coccidia, 3% by enterohelminths. 90.3% of calves aged from three to six months were parasitised by coccidia, 7.4% by enterohelminths. Young cattle, aged from 6 to 12 months, were parasitised in 94.6% by coccidia and in 14.1% by enterohelminths. 86.6% of pregnant heifers at the age of 14 to 19 months had coccidia, 7.7% enterohelminths. The total farm capacity was about 950 animals. Throughout the year the number of examined animals amounted to 1,195, out of this number 958 animals, i. e. 80.2%, were infested by coccidia, 55.6% of animals by the species Eimeria bovis, 47.0% by E. zuernii, 43.3% by E. auburnensis, 39.4% by E. ellipsoidalis, 25.2% by E. cylindrica, 13.6% by E. subspherica, 3.1% by E. bukidnonensis, 1.7% by Isospora spp., 0.4% by E. brasiliensis, 0.08% by E. pellita. Enterohelminths were observed only in 96 animals, i. e. in 8.03%. Trichocephalus was found in 3.9% of animals, Cooperia in 2.4%, Ostertagia in 0.6%, Chabertia in 0.4%, Nematodirus in 0.3%, Capillaria in 0.2%, Oesophagostomum in 0.2%, Bunostomum in 0.1% and Trichostrongylus in 0.1%. The highest elimination of oocysts of coccidia was observed in August, January and February, of eggs of enterohelminths from August to October. In all age groups the most frequent was the occurrence of pathogenic species of coccidia.     

The survival ability of salmonella, coccidia oocysts and ascarid eggs in laying hen feces from different housing systems]

The time of survival of Salmonella typhimurium, coccidia oocysts and ascaris eggs in manure of layer was determined in 5 different housing systems and 2 storing places for litter. The experiments were carried out in a stable of experimental station Frankenforst of the university of Bonn with a flock of 2200 hens. The effects of the environment conditions temperature, dry matter content, pH-value and intestinal microflora of the manure have also been studied. The time of survival was different depending on the housing system. A recovery of viable coccidia oocysts was possible after 13-370 days, ascaris eggs 53-347 days and Salmonella typhimurium 2-175 days. The tenacity of the investigated test organism mainly depend on the dry matter content of the manure. The longest period of survival of salmonellas was found in dry environment conditions, were as coccidia oocysts and ascaris eggs have been observed with the shortest period of survival. The possibility of the examined resistant parasite stages to develop was disturbed. Only few of them were able to develop and with a longer development time than those examined in the control suspension. The results of this study indicate that chicken manure, before using it in plant production, should be stored long enough to prevent men or animals from possible infections.     

鸡球虫病药物防治研究进展

鸡球虫病是严重危害养鸡业的寄生虫病之一.长期以来,鸡球虫病主要依靠抗生素类、化学合成药类和中草药等药物进行防治,由于球虫极易产生耐药性,给药物防治带来了困难.文章从鸡球虫生物学特性、防治鸡球虫病的药物及鸡球虫病的药物防治方法等方面做一综述.     

鸡球虫耐药性研究进展

到目前为止，鸡球虫病的防治仍以药物控制为主，大量抗球虫药的长期使用，导致了鸡球虫耐药性的产生，给鸡球虫病的防治带来很大的困难。本文总结近年来国内外学者研究的成果，在鸡球虫耐药性概述、抗球虫药及其作用机制、耐药性产生的机制、耐药性的特点、耐药性的检测方法、耐药性的获得及敏感性恢复，耐药性的延缓或解决措施等方面进行了综述，提出对鸡球虫耐药性的展望，为消除耐药性，更好地防治鸡球虫病提供依据。