纯中草药散剂对鸡球虫病的疗效试验

250只海蓝灰雏鸡，饲养至15日龄，随机分为5组，分别为试验组（1％纯中药散剂组、0.5％纯中药散剂组、0.25％纯中药散剂组）、阳性对照组（只感染球虫卵囊不给药）及药物（三字球虫粉）对照组，每组50只。试验鸡全部经口感染鸡柔嫩艾美耳球虫孢子化卵囊，试验组及药物对照组连续给药5 d。在第8 d将鸡全部扑杀剖检。试验期间统计每组鸡的死亡率、增重率、粪便记分、病变记分、病变值、卵囊计数、卵囊比数、卵囊值、鸡抗球虫指数等指标。结果表明：0.5％～1％的纯中药散剂抗球虫指数在160～180之间，为中效抗球虫药。     

禽源生物制品生产中蛋传性病原的控制方法

1 禽用活疫苗外源病毒检测 《中华人民共和国兽药典》(2010年版,三部)对活疫苗外源病毒检测的要求: 1.1 样品处理 取样品2～3瓶,对毒种或活疫苗进行检验时,将样品混合,用相应有特异性抗血清中和后作为检品(另有规定除外);对细胞进行检验时,经3次冻融后混合作为检品;用鸡检查法检验时,样品不处理.     

3种中药对鸡球虫病四价活疫苗卵囊繁殖的影响

正鸡球虫病是我国养鸡生产中常见且危害大的疾病。近年来,由于球虫耐药性问题以及严峻的食品安全形势,采用疫苗代替抗球虫药来预防鸡球虫病的人越来越多。然而,目前的鸡球虫病疫苗均为活卵囊疫苗,免疫接种后需在鸡体发育繁殖,刺激机体免疫应答。所以,在小鸡阶段鸡体建立抗球虫免疫力期间,要避免使用抗球虫药和广谱抗菌抗球虫类药物,比如磺胺类、四环素类药物。为了解一些中药对鸡球虫病活疫苗卵囊繁殖的影响,特此进行本试验,以便为接种球虫病疫苗的鸡群选用药物提供依据。1材料与方法1.1供试药物     

玉溪市一例散养土杂鸡球虫病的虫种鉴定

为了对云南省玉溪市某散养土杂鸡场一例球虫病例进行虫种鉴定，本试验通过发病情况调查、病理剖检、虫卵检查、虫卵计数、特异性引物PCR扩增、18S rRNA基因序列测序等方法进行检测分析。结果显示：病鸡盲肠粗大肿胀，肠腔内有棕黑色黏液样内容物；虫卵大小在22.922～26.621μm×17.433～19.631μm之间，每个孢子化卵囊含4个孢子囊，每个孢子囊含有2个子孢子，符合柔嫩艾美耳球虫特征；柔嫩艾美耳球虫特异性基因检测为阳性；该虫种18S rRNA基因序列与柔嫩艾美耳球虫序列的同源性最高为99.61%～99.92%，且在进化树上构成一个分支；虫卵计数卵囊数（OPG）为1.7×10⁴。试验表明该鸡场鸡群感染了柔嫩艾美耳球虫，且为中度感染；用磺胺类药物加祛湿中药治疗，疗效显著。     

不同日龄雏鸡对柔嫩艾美球虫早熟株卵囊的易感性及免疫力

对１,７和２５日龄鸡分别感染１０００个早熟柔嫩艾美球虫孢子化卵囊、感染１２０ｈ后,每２４ｈ收集粪便１次,进行卵脏数,连续７ｄ,３种日龄鸡排卵囊总数分别为４６０．９,８０２．０和１４１７．７万个／只。感染后１４ｄ对７日龄鸡的免疫组及相对应的２个不免疫对照组每只鸡攻击１００个柔嫩艾美球虫亲本株孢子化卵囊。攻毒１４４ｈ后,每２４ｈ收集粪便１次,进行卵囊计数,连续５ｄ,各组鸡的囊总量分别为４１．４,５．８     

白头翁不同提取物及复方抗鸡柔嫩艾美耳球虫研究

以存活率、相对增重率、盲肠病变记分、卵囊值和抗球虫指数为判定指标,考察白头翁不同提取物及复方对鸡柔嫩艾美耳球虫孢子化卵囊致病性的影响(体外试验)和抗鸡柔嫩艾美耳球虫感染效果(体内试验)。体内、体外试验结果均表明,白头翁不同提取物及复方均能提高相对增重率,降低盲肠病变计分和卵囊值。抗球虫指数(ACI)检测表明,白头翁不同提取物及复方对球虫孢子化卵囊有微弱的抑杀效果,对鸡柔嫩艾美耳球虫感染预防效果较差,不宜作为鸡柔嫩艾美耳球虫病的预防和治疗药物。     

不同品种地方鸡对柔嫩艾美球虫的易感性分析

按每只鸡12×10~4个孢子化卵囊的剂量,分别感染20日龄大骨鸡、鹿苑鸡、油鸡和固始鸡,以存活率、相对增重率、血便扣分、病变扣分、卵囊扣分和抗球虫指数为评价指标,分析比较4个地方品种鸡对柔嫩艾美球虫的易感性。试验重复1次。结果显示,4个地方品种鸡对柔嫩艾美球虫的易感性不同。固始鸡对柔嫩艾美球虫易感性最高,表现为感染后的死亡率最高,病变扣分最多,抗球虫指数也最低。油鸡对柔嫩艾美球虫的易感性最低,表现为感染后的死亡率为零,病变扣分和血粪扣分最少,每鸡平均排卵囊数最少,抗球虫指数最高。大骨鸡和鹿苑鸡对柔嫩艾美球虫的易感性相近,位于固始鸡与油鸡之间。     

毒害艾美耳球虫未孢子化卵囊壁的分离与纯化

用毒害艾美耳球虫孢子化卵囊接种50只15日龄无球虫感染的雏鸡,感染后第6~12天,每隔12 h收集1次粪便,按常规方法分离卵囊。未孢子化卵囊经次氯酸钠溶液处理后,用1.1 mol/L蔗糖溶液漂浮法纯化卵囊。采用玻璃珠涡旋法破碎卵囊壁,最后用孔径10.0μm的聚对苯二甲酸乙二醇酯(PET)膜过滤获得纯化的卵囊壁。研究结果为进一步研究毒害艾美耳球虫卵囊壁的结构及其蛋白组成奠定了基础。     

鸡球虫病活疫苗实验室安全性试验

鸡球虫病不仅影响鸡健康状况、生长发育以及死亡率，而且也影响着鸡规模化养殖的健康持续发展。目前养殖场主要通过抗生素药物治疗球虫病，导致该病耐药治疗效果差，且容易出现药物残留风险，影响鸡肉品质。目前鸡可以接种鸡球虫活疫苗，激发鸡免疫器官产生抗体，减少疾病的危害。为了研究鸡球虫活疫苗的安全性，采用一次单剂量接种、单剂量重复接种和一次超剂量接种的试验，通过免疫后的临床症状、解剖后病变情况、组织切片镜检等方法，来验证鸡球虫活疫苗的安全性。本文主要概述了鸡球虫病的生物学特性、流行规律、鸡临床特征以及鸡球虫病活疫苗安全性试验的方法。简述目前鸡球虫活疫苗安全性实验得出的结果进展。     

吉林市城区鸡球虫病的流行病学调查

通过对吉林城区具有代表性的鸡场进行鸡球虫病的感染情况调查,采集鸡新鲜粪便,用饱和盐水漂浮法收集卵囊,制片镜检,观察卵囊和孢子化卵囊的形状、颜色,测量卵囊的大小,鉴定出艾美耳属球虫3种,即柔嫩艾美耳球虫、堆型艾美耳球虫、巨型艾美耳球虫,其中柔嫩艾美耳球虫为优势种,其主要侵害鸡的盲肠,导致贫血、消瘦甚至死亡,发病多为15～40日龄的雏鸡,为有效预防鸡球虫病提供参考.     

Success and failure: The role of relative humidity levels and environmental management in liveEimeria vaccination of cage-reared replacement layer pullets

Contrary to the traditional view that layer flocks housed in conventional cages are unlikely to suffer coccidiosis caused byEimeria species, this enteric disease has become an emergent issue. Coccidiosis outbreaks in layers are frequently associated with the failure to develop protective immunity at a young age. Layer hens housed in cages are usually sourced from replacement layer pullets housed in similar cages in the rearing barn. Live coccidiosis vaccines administered to young chicks provide a small dose of vaccine oocysts that infect and replicate within the vaccinated birds, resulting in the release of progeny oocysts into the rearing environment; these oocysts must become infective (i.e., sporulate) and then reinfect the partially immune birds in order for the vaccine to generate complete protective immunity. Three factors are needed for successful oocyst sporulation: temperature (4 to 37°C; optimal 29°C), oxygen access, and adequate RH. The purpose of this study was to assess the degree of protective immunity elicited by birds spray-vaccinated with a live coccidiosis vaccine at the hatchery. During the critical period for oocyst sporulation and cycling, RH levels decreased to 13 to 19%, resulting in inadequate oocyst cycling and minimal protection against homologous challenge at 6 and 12 wk of age. This vaccination failure highlights the need to monitor RH in the barn and modify the barn environment so that conditions promote, rather than impede, the success of live coccidiosis vaccines.     

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。     

Coccidiosis immunization: effects of mushroom and herb polysaccharides on immune responses of chickens infected with Eimeria tenella

An experiment was conducted to investigate the effects of polysaccharide extracts (E) of two mushrooms, Lentinus edodes (LenE) and Tremella fuciformis (TreE), and an herb, Astragalus membranaceus (AstE), on the immune responses of chickens infected with Eimeria tenella. A total of 180 broiler chickens were assigned to nine groups: three groups were fed with each of the extracts (LenE, TreE, and AstE), three groups were fed with the extracts and immunized with live oocyst vaccine (LenE+V, TreE+V, and AstE+V), a group was immunized with the vaccine only, and there were two controls (E. tenella-infected and noninfected groups). The oocyst vaccine was given at 4 days of age, and the extracts (1 g/kg of the diet) were supplemented from 8 to 14 days of age. At 18 days of age, all birds except those in the noninfected group were infected with 9 x 10(4) sporulated oocysts. The results showed that at 7 days postinfection (p.i.), birds fed the extracts without vaccination had lower body weight (BW) gain than those given the vaccine only. However, the extracts in conjunction with the vaccine significantly enhanced BW gain of the infected chickens compared with the vaccine group. Of the three extracts, LenE and TreE showed a better growth-promoting effect. The extracts largely increased oocyst excretion of droppings during the primary response postvaccination. The cecal peak oocyst output and lesion scores measured at 7 days p.i. were higher in the groups fed the extracts than in the group immunized with the vaccine only, whereas those of the groups fed with the extracts and immunized with the vaccine were not significantly different from the vaccine group. Of the three extracts, both LenE- and AstE-fed groups showed lower cecal oocyst output. Thus, as compared with the extracts, the live, attenuated vaccine showed better results with significantly increased immune response in coccidial infected birds. The polysaccharide extracts may prove useful against avian coccidiosis, and, particularly when they are used in conjunction with vaccine, they have shown preliminary promise against the experimental coccidial infection.     

鸡球虫重组活载体疫苗研究进展

对细菌和病毒为载体的鸡球虫重组活载体疫苗研究进行了综述,旨在为鸡球虫病的免疫预防提供新思路和参考.     

紫茎泽兰处理鸡柔嫩艾美耳球虫后差异基因筛选和分析

为获得紫茎泽兰处理鸡柔嫩艾美耳球虫后差异基因,将0.5%紫茎泽兰提取液作用于鸡柔嫩艾美耳球虫卵囊孢子化过程,采用抑制消减杂交技术（SSH）筛选处理后卵囊的差异表达基因,通过GO和COG功能预测分析,并采用实时荧光定量PCR验证差异表达的基因。结果显示,采用SSH成功构建了紫茎泽兰处理前后鸡柔嫩艾美耳球虫卵囊的cDNA消减文库,获得86条ESTs序列,经拼接和聚类后得到31条独立基因（Unigenes）,其中有23个基因有功能注释,8个基因没有功能注释,另外有4个基因没有同源性匹配。为进一步验证文库的特异性,从中随机选取3个差异表达基因,运用实时荧光定量PCR技术验证表面抗原13、3-羟酰辅酶A脱氢酶和细胞色素P450基因在紫茎泽兰处理前后的表达差异,结果显示,3个基因在经紫茎泽兰处理的鸡球虫孢子化卵囊中的表达量明显低于未处理组,说明紫茎泽兰对柔嫩艾美耳球虫卵囊具有一定的活性抑制作用。本试验获取了紫茎泽兰作用柔嫩艾美耳球虫卵囊的主要调控基因,为将紫茎泽兰研发成环境杀虫剂奠定了良好的理论基础,也可为球虫致弱疫苗或基因缺失疫苗的靶标筛选研究提供参考。     

Selection and characterisation of two attenuated vaccine lines of Eimeria tenella in Australia

OBJECTIVE: To attenuate two strains of Eimeria tenella by selecting for precocious development and evaluate the strains in characterisation trials and by field evaluation, to choose one precocious line for incorporation into an Australian live coccidiosis vaccine for poultry. DESIGN: Two strains from non-commercial flocks were passaged through chickens while selecting for precocious development. Each strain was characterised for drug sensitivity, pathogenicity, protection against homologous and heterologous challenge, and oocyst output in replicated experiments in which the experimental unit was a cage of three birds. Oocyst output and/or body weight gain data collected over a 10 to 12 day period following final inoculation were measured. Feed conversion ratios were also calculated where possible. RESULTS: Fifteen passages resulted in prepatent periods reduced by 24 h for the Redlands strain (from 144 h to 120 h) and 23 h for the Darryl strain (from 139 h to 116 h). Characterisation trials demonstrated that each precocious line was significantly less pathogenic than its parent strain and each effectively induced immunity that protected chickens against challenge with both the parent strain and other virulent field strains. Both lines had oocyst outputs that, although significantly reduced relative to the parent strains, remained sufficiently high for commercial vaccine production, and both showed susceptibility to coccidiostats. CONCLUSION: Two attenuated lines have been produced that exhibit the appropriate characteristics for use in an Australian live coccidiosis vaccine.     

In vitro destruction of Eimeria oocysts by essential oils

This study aims to assess the ability of essential oils (EOs) to destroy Eimeria oocyst in vitro using microscopic counting and 273 nm absorbing material release. A screening for the ability of ten EOs to destroy Eimeria oocyst was carried out in liquid medium. Out of these ten, artemisia, tea tree, thyme and clove EOs were identified as being the most effective. The treatment of Eimeria oocyst with these EOs leads to their lysis as shown by the release of substances absorbing at 273 nm. These results were obtained after approximately three hours contact. Four EOs were proven to destroy Eimeria oocysts in a few hours at low concentration. This destructive effect is a consequence of their lysis. This work is a preliminary contribution aiming to develop a new generation of natural efficient agents for destroying Eimeria oocyst to fight coccidiosis in broiler chicken.     

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.     

Use of live oocyst vaccine in the control of turkey coccidiosis: Effect on performance and intestinal morphology

An experiment was conducted to determine the effects of different coccidiosis-preventing programs on performance and intestinal morphology of commercial turkeys. Three hundred fifteen1-d-old female commercial cross turkey poults (British United Turkeys, BUT Big 9) were distributed into 3 treatments with 5 replicates of 21 birds each. Three programs were evaluated from 1 to 70 d of age, where program 1 had no anticoccidial drug and no vaccination against coccidiosis; program 2 had an anticoccidial drug (maduramycin 1%, 5 ppm); and program 3 had a vaccination (commercial vaccine, 4 species of Eimeria). All the groups were challenged with a dose of oocysts sporulated (20,000/bird) of 2 species of Eimeria at 21 d of age. In the growing phase (d 0–28), BW, BW gain, and FCR were significantly greater in treated groups compared with control group. In the fattening phase, the performance was not affected by treatments. Treatments and coccidiosis challenge had no significant effects on intestinal villus height. These observations support other reports that confirm live oocyst vaccination can be used effectively as a preventive against avian coccidiosis in commercially reared turkeys.     

In vivo effects of CpG oligodeoxynucleotide on Eimeria infection in chickens

Poultry coccidiosis is the major parasitic disease of poultry and, until now, no recombinant vaccine has been developed. Short oligodeoxynucleotides (ODNs) containing unmethylated CpG motifs (CpG ODNs) have been shown to be effective immunoprotective agents and vaccine adjuvants in mammalian systems. Their use in poultry to protect against intracellular parasites has not been reported to date. The present work investigated the effects of CpG ODN treatment on host susceptibility to Eimeria infection in two chicken strains with different genetic background, SC and TK. The data show that CpG ODN enhanced the birds' resistance to coccidiosis in a normally susceptible chicken strain (TK), as shown by reduced oocyst shedding and improved weight gain. CpG treatment had a differential effect on body weight gains and serum antibody responses, depending on the chicken strain and ODN dose, delivery route, and backbone. This study shows for the first time that CpG ODNs could be used as immunoprotective agents in Eimeria-infected chickens to enhance resistance to the pathogen and improve performance. Future research is needed to optimize their use alone and as vaccine adjuvants that may lead to better and more efficient vaccine applications.