重庆市江津地区鸡球虫种类调查

对重庆江津地区 12个养鸡场和 18个养鸡户的鸡球虫感染情况进行调查 ,调查结果显示 ,养鸡场鸡球虫感染多为混合感染 ,一般有 4种以上的艾美耳球虫。本次共检出有 7种艾美耳球虫 ,即 :柔嫩艾美耳球虫 (Eimeriatenella) ,巨型艾美耳球虫 (E.maxia) ,堆形艾美耳球虫 (E.acervulina) ,毒害艾美耳球虫 (E.necatrix) ,布氏艾美耳球虫 (E.brunetti) ,变位艾美耳球虫 (E.mivati)和和缓艾美耳球虫 (E.mitis) ,其致病虫种主要为柔嫩艾美耳球虫、堆型艾美耳球虫和巨型艾美耳球虫     

鸡柔嫩艾美球虫DNA疫苗pcDNA4．0（C）-pEtK2-IL-2对其他球虫的交叉保护力

将柔嫩艾美球虫DNA疫苗pcDNA4．0（c）-pEtK2-IL-2以50μg分别对14日龄和21日龄雏鸡进行胸部肌肉注射免疫，28日龄对各未免疫组与免疫组鸡分别口服接种柔嫩艾美球虫、巨型艾美球虫、堆形艾美球虫和毒害艾美球虫孢子化卵囊，36日龄剖杀，分析比较免疫保护效果。结果显示，柔嫩艾美球虫攻毒免疫试验组与柔嫩艾美球虫攻毒未免疫对照组比较，增重、盲肠病变记分、OPG值差异显著（P〈0．05），免疫保护效果明显，抗球虫指数（ACI）达186．50；而巨型艾美球虫、堆形艾美球虫和毒害艾关球虫攻毒免疫试验组的ACI分别为147．85、142．71和153．82，增重、盲肠病变记分、OPG值和ACI与相应的攻毒未免疫对照组比较，均有不同程度改善，免疫保护效果不明显。表明，该DNA疫苗对柔嫩艾美球虫的攻击具有完全免疫保护作用，而对巨型艾美球虫、堆形艾关球虫、毒害艾美球虫的攻击具有部分交叉保护力。     

秦皇岛地区十个鸡场鸡球虫种类的初步调查

通过对秦皇岛地区10个养鸡场鸡球虫种类的调查，发现8个养鸡场为球虫阳性，共检出6种球虫，分别是哈氏美耳球虫（34％）、柔嫩艾美球虫（26％）、早熟艾美耳球虫（12％）、和缓艾美耳球虫（12％）、堆形艾美耳球虫（11％）和毒害艾美耳球虫（5％）。     

首个国产鸡球虫疫苗问世保护率超95％

佛山市正典生物技术有限公司自主研发的“鸡球虫病四价活疫苗（柔嫩艾美耳球虫PTMZ株＋毒害艾美耳球虫PNHZ株＋巨型艾美耳球虫PMHY株＋堆型艾美耳球虫PAHY株）”近日获得国家三类新兽药证书，这是国内首个全部采用早熟选育致弱的药物敏感虫株组成的鸡球虫病四价活疫苗。     

湖南省衡阳市鸡球虫病病原种类与流行情况调查

本文通过地衡阳市辖区县市区鸡球虫感染的调查研究，结果表明，衡阳市鸡球虫感染严重，鸡群感染率为56．2％，尤以雏鸡感染严重，农户放养鸡感染率高达72．8％，专业户饲养鸡高达63．5％，青、成年鸡农户放养、专业户饲养的感染率分别为37．5％，30．85，经实验室鉴定，共查出柔嫩艾美耳球虫、毒害艾美耳球虫、巨型艾美耳球虫、早熟艾美耳球虫、堆型艾美耳球虫、和缓艾美耳球虫6个种，其中柔嫩艾美耳球虫为优势种，检出率达94．6％。，调查还表明衡阳市鸡球虫发病主要有4－6月份的梅雨季节，鸡日龄、饲养管理水平，不同的饲养环境条件等与鸡球虫感染关系密切。     

闽北麻鸡暴发鸡球虫病的诊治报告

鸡球虫病是养禽业中常见的一种原虫病。主要危害3月龄以内鸡,15～50日龄时更为常见且病情较严重。除引起死亡外,病愈的鸡生长、发育及增重受到严重影响。寄生于鸡的球虫种类很多,致病力最强的为柔嫩艾美尔球虫(寄生于盲肠)、毒害艾美尔球虫、堆形艾美耳球虫和巨型艾美耳球虫(寄     

鸡球虫病的防治方法

鸡球虫病的病原为原虫中艾美耳科艾美耳属的球虫,世界各国已经记载的鸡球虫种类共有13种之多,我国已发现柔嫩、毒害、巨型、堆型等9种艾美耳球虫,其中致病力最强的是柔嫩艾美耳球虫和毒害艾关耳球虫。     

1例鸡球虫病的诊治与病因分析

鸡球虫病是养禽生产中最常见的一种寄生性原虫病,由艾美耳属多种球虫寄生于鸡的肠上皮细胞内所引起,感染鸡的球虫有7种,分别为柔嫩、毒害、巨型、堆型、布氏、和缓和早熟艾美耳球虫。以柔嫩艾美耳球虫和毒害艾美耳球虫致病力最强,     

动物医院门诊病鸡球虫感染情况及其种类调查

对送检扬州大学动物医院50只鸡的球虫感染情况调查显示，15日龄以内鸡的球虫感染率为20％，15—50日龄鸡为96．5％，50日龄以上鸡为100％，总感染率为90％。共检出7种球虫，即柔嫩艾美耳76％、堆型艾美耳56％、巨型艾美耳22％、早熟艾美耳16％、和缓艾美耳10％、毒害艾美耳4％和布氏艾美耳2％。柔嫩和堆型为优势种。球虫多为混合感染的占71．1％，2种球虫混合感染最多占44.4％。传染性法氏囊病例鸡中，严重感染球虫的最多为43．1％。     

堆形艾美耳球虫早熟株选育及相关生物学特性研究

为了选育堆形艾美耳球虫（Eimeria acervulina）早熟株及观测其生物学特性,运用Jeffers创立的艾美耳属球虫早熟株选育方法,对实验室保存的堆形艾美耳球虫进行连续传代早熟选育,并对获得的早熟株与母株孢子化卵囊大小、繁殖能力、致病性以及免疫保护力等指标进行了测定,比较堆形艾美耳球虫早熟株与母株之间的基本生物学特性差异。经过18代早熟选育后,堆形艾美耳球虫的潜在期由母株的99h缩短至84h。与母株相比,早熟株的孢子化卵囊大小明显减小（P〈0．05）,卵囊繁殖能力降低了17％～42%。致病性试验中,早熟株感染组的平均增重均高于母株,而病变记分相当。免疫保护试验中,早熟株与母株的免疫保护力相当,其卵囊抑制率略低于母株,差异不明显（P〉0．05）。结果表明,本研究成功选育出堆形艾关耳球虫早熟株,对母株攻击具有保护力,为鸡球虫疫苗的研制奠定了基础。     

Inter- and intra-strain variation and PCR detection of the internal transcribed spacer 1 (ITS-1) sequences of Australian isolates of Eimeria species from chickens

The objective of this study was to confirm the presence of seven species of Eimeria involved in chicken coccidiosis in Australia by comparing internal transcribed spacer 1 (ITS-1) sequences, ITS-1 polymerase chain reaction (PCR) methods and to apply phylogenetic analysis to assess evolutionary relationships of Australian isolates. Twenty-two distinct ITS-1 regions of 15 Australian Eimeria isolates were sequenced, and analysed using maximum parsimony, distance and maximum likelihood methods. Poor bootstrap support, resulting from high ITS-1 sequence heterogeneity between all species groups, resulted in polychotomy of the Eimeria species in all three trees generated by these analyses. Percentage identity analyses revealed two distant ITS-1 lineages in both E. mitis and E. maxima at the same levels that separate the two species E. tenella and E. necatrix. One E. maxima lineage consisted of Australian isolates, the other American isolates, with one European sequence (originating from the same isolate) in each lineage. One Australian E. praecox sequence was only distantly related (33% variation) to three E. praecox sequences from Australian and European isolates. Short and long ITS-1 variants were isolated from both E. tenella (cloned line) and E. necatrix isolates with deletions (106 and 73 bp, respectively) in the short variants within the 3' region of the ITS-1 sequence. ITS-1 sequences of strains of both E. brunetti and E. acervulina species varied the least. Apart from E. maxima, all of the ITS-1 sequences of the six remaining individual species clustered to the exclusion of other species in all phylogenetic trees. Published ITS-1 tests for E. necatrix, E. acervulina, E. brunetti and E. tenella, combined with three new tests for E. mitis, E. praecox and Australian E. maxima amplified all respective Australian isolates specifically in a nested format using conserved ITS-1 PCR products as template to improve the sensitivity. All PCR tests were confirmed against a collection of 24 Australian chicken Eimeria isolates and contaminating species were detected in some instances. In conclusion, once the genetic variation between species and strains is determined, the ITS-1 is a good target for the development of species-specific assays, but the ITS-1 sequences alone do not seem suitable for the confirmation of phylogenetic inferences for these species. This study reports the first attempt at the analysis of the phylogeny and sequence comparison of the Eimeria species involved in chicken coccidiosis in Australia.     

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.     

Cloning and nucleotide sequencing of the second internal transcribed spacer of ribosomal DNA for three species of Eimeria from chickens in Taiwan

Coccidiosis of chickens caused by protozoan parasites of the genus Eimeria (Coccidia: Eimeriidae) is an enteric disease that results in great economic losses throughout the world, including Taiwan. Using polymerase chain reaction (PCR) with primers specific for the second internal transcribed spacer (ITS-2) of ribosomal DNA (rDNA), three species of Eimeria, E. tenella, E. maxima, and E. acervulina have been successfully characterised from chickens in Taiwan. The sizes of PCR products from various isolates representing these three species were between 370 and 580 base pairs (bp). After cloning and sequencing of the PCR products, high nucleotide sequence identity (96.8-100%) was observed within a species. In addition, ITS-2 nucleotide sequences for E. tenella had higher homology (98.5-99.3%) than E. maxima (81.6-96.5%) when compared with appropriate sequences deposited in GenBank. To our knowledge, this is the first report of a 412-bp ITS-2 sequence for E. acervulina from chickens.     

特异PCR方法用于鸡的3种艾美耳球虫混合感染鉴别的研究

用单卵囊分离法获得的鸡的3种艾美耳球虫（每种各2株）卵囊：柔嫩艾美耳球虫（Eimeria tenella）、巨型艾美耳球虫（E．maxima）、堆型艾美耳球虫（E．acervulina）。经纯化、提取基因组DNA后，用报道的种特异引物做PCR扩增分析，以确定是否为纯种。结果发现这3种球虫均存在混合感染的情况。该结果为进一步研究这3种球虫奠定了基础，并说明特异PCR方法能够有效地、快速地鉴别球虫虫种。     

Improved polymerase chain reaction technique for determining the species composition of Eimeria in poultry litter

An improved polymerase chain reaction (PCR)-based method for determining the species composition of Eimeria in poultry litter was developed by incorporating species-specific internal standards in the assay. Internal standard molecules were prepared by fusing seven different Eimeria species-specific intervening transcribed sequence 1 (ITS1) rDNA primer pairs to a non-Eimeria DNA molecule and by cloning the hybrid DNA molecules into a plasmid. The internal DNA standards were then used in Eimeria-specific ITS 1 PCR, and they were found to be capable of detecting E. acervulina, E. maxima, E. praecox, and E. tenella oocysts isolated directly from poultry litter.     

Differential diagnosis of five avian Eimeria species by polymerase chain reaction using primers derived from the internal transcribed spacer 1 (ITS-1) sequence

Chicken coccidia are protozoan parasites of the genus Eimeria. They cause economical losses in the poultry industry globally. The various species can be distinguished on the basis of the morphology of the oocysts and parasitic site in intestine, but these criteria sometimes are unreliable. Therefore, a species-specific polymerase chain reaction (PCR) was developed. Based on variable sequence regions, specific primers were constructed for the differentiation of five Eimeria species (Eimeria acervulina, E. brunette, E. maxima, E. necatrix, and E. tenella). PCR products were amplified from coccidian vaccine (coccivac-D and coccivac-B) and E. tenella and were subsequently sequenced. Similarities of the five species sequences between the vaccines and Genbank were 94-100%. Analysis of the E. tenella internal transcribed spacer 1 (ITS-1) partial sequence from Taiwan and from Genbank indicated that the similarity was 99.6%. The PCR sensitivity test of E. tenella in Taiwan is 50 oocysts. The five sets of primers will not amplify any non-specific bands of the chicken genome or its intestinal contents. Therefore, the five sets of specifically designed primers are guaranteed to be useful for differential diagnosis of avian coccidiosis caused by Eimeria spp.     

Antigenic variation among strains of Eimeria maxima and E. tenella of the chicken.

The immunogenicity and cross-protection of Eimeria maxima or E. tenella in chickens against their homologous and heterologous strains were evaluated in two experiments. The immunizing strain of E. tenella protected against itself and partially against heterologous strains. The North Carolina (NC) strains of E. tenella were more virulent than the Delmarva (DMV) strains. Growth of the unimmunized groups was depressed 66% and 32% for the NC and DMV strains, respectively. Growth of the immunized-challenged groups (DMV and NC) was depressed by 13%. The DMV E. maxima strains were more virulent than the NC strains. Growth of the unimmunized challenged groups was depressed by 47% (DMV) and 13% (NC). Results demonstrated that there are antigenic variations among strains of two species of chicken coccidia.     

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.     

Diagnosis of Eimeria species using traditional and molecular methods in field studies

The objective of this study was to identify and characterize species of Eimeria in broiler chickens using traditional morphological and pathological plus molecular (DNA amplification) diagnostic methodologies. Using a combination of those techniques it was possible to identify the presence of multiple circulating species in the flock as well as higher frequencies for some of them, especially Eimeria praecox and Eimeria maxima, which were identified in 100% of the flocks. The frequencies of the other species were Eimeria mitis and Eimeria necatrix (93.3%), Eimeria tenella (76,7%), Eimeria acervulina (56.7%) and Eimeria brunetti (16.7%). However using the lesion score, the most common species were E. maxima (46.7%), E. acervulina (30%), E. tenella (23.3%), and E. necatrix (10%). E. brunetti and E. praecox were not identified by using lesion score. DNA amplification had detection sensitivity for Eimeria species in the field samples of at least 20 oocysts. The implementation of DNA amplification as a routine diagnostic technique in aviaries can assist Eimeria population.     

野生乌苏里貉18SrDNA全序列的克隆测序及系统进化树分析

试验采用聚合酶链式反应方法,从野生乌苏里貉耳部肉中提取DNA,扩增出18S rDNA的全序列,并对其进行克隆测序,获得长度为1831 bp的序列,并将序列提交到GenBank上,经BLAST分析结果表明,测序得到的乌苏里貉18S rDNA与其他物种同源性均在90%以上,说明18S rDNA基因具有较高的保守性,可以作为分类学当中系统进化分析的重要参考依据。