安徽合肥市鸡球虫种类及感染情况调查

本文采用饱和盐水漂浮、重铬酸钾培养等方法，对合肥市79份鸡的粪样进行了鸡球虫感染情况的检查。结果表明：鸡球虫感染率为62．03％(49／79)。经鉴定，所获7种球虫均隶属艾美耳属，即：毒害艾美耳球虫(Eimeria 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)、和缓支美耳球虫(E．mitis)、哈氏艾美耳球虫(E．hagani)和早熟艾美耳球虫(E．praecox)。文中还对鸡球虫与地区、日龄的关系及优势虫种等进行了分析，得出合肥市鸡球虫感染率存在一定的地区性和日龄差异，鸡球虫优势虫种为毒害艾美耳球虫和堆型艾美耳球虫。     

上海市鸽球虫感染情况初步调查与种类鉴定

为了调查上海市鸽球虫的感染情况和种类,分别采集奉贤区肉鸽场和信鸽场的110份和44份粪便,以及南汇区肉鸽场的100份粪样,采用麦氏虫卵计数法对粪便球虫卵囊进行计数,收集阳性粪便中的卵囊,经孢子化后进行种类鉴定。结果显示,上海市鸽球虫的平均感染率为52.8%(134/254),平均每克粪便卵囊数(oocyst per gram,OPG)为50 159个,为中度感染。肉鸽感染率(55.2%)和平均OPG(52 110个)均高于信鸽(40.9%和37 583个),奉贤肉鸽感染率(60.9%)高于南汇(49.0%),但南汇肉鸽的OPG(88 634个)明显高于奉贤(25 400个)。从肉鸽阳性粪便中鉴定出5种球虫,即拉氏艾美耳球虫(Eimerialabbeana)、鸽艾美耳球虫(E.columbae)、原鸽艾美耳球虫(E.columbarum)、杜氏艾美耳球虫(E.duculai)以及卡氏艾美耳球虫(E.kapotei),拉氏艾美耳球虫为优势种。在信鸽阳性粪便中鉴定出4种球虫,即拉氏艾美耳球虫、杜氏艾美耳球虫、鸽艾美耳球虫以及原鸽艾美耳球虫,原鸽艾美耳球虫为优势种。本次调查表明上海市肉鸽和信鸽的球虫感染率和粪便OPG均偏高,调查结果为及时做好鸽球虫病的防治提供了基础数据。     

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。     

青海省祁连县绵羊球虫感染情况和种类调查研究

对青海省祁连县默勒镇96只绵羊进行了球虫感染情况和种类调查研究。结果显示:总感染率为70.8%,其中1岁绵羊感染率83.3%,2岁羊感染率76.7%,成年羊感染率46.7%;多为2～5种球虫混和感染;平均OPG值为154.1(20～1460)。显微镜下对孢子化卵囊进行形态学观察,测量卵囊大小,显微照相,并列出了各种球虫的主要鉴别特征,绘制了卵囊形态图,进行虫种鉴定,共检出12种艾美耳球虫,其中确定的有11种:小型艾美耳球虫(Eimeria.parva)、类绵羊艾美耳球虫(E.ovinoidalis)、槌形艾美耳球虫(E.crandallis)、威布里吉艾美耳球虫(E.weybridgensis)、苍白艾美耳球虫(E.pallida)、阿撒他艾美耳球虫(E.ahsata)、浮氏艾美耳球虫(E.faurei)、卵状艾美耳球虫(E.oodeus)、巴库艾美耳球虫(E.bakuensis)、颗粒艾美耳球虫(E.granulosa)以及错乱艾美耳球虫(E.intricata),前4种为优势虫种;未定种一种。     

宁夏四地区奶牛球虫病流行病学调查及虫种鉴定

为了掌握宁夏回族自治区四地区奶牛球虫的感染情况及优势流行虫种，本试验对石嘴山、银川、吴忠和青铜峡4个市的14个规模化奶牛场的球虫感染情况进行调查，按照每个月龄奶牛总数的15%比例随机进行样本采集并用显微镜检测球虫。结果显示：采集的1 938份粪样中，856份为阳性，平均感染率为44.17%;共发现12种艾美耳球虫(Eimeria),分别是牛艾美耳球虫(E.bovis)、柱状艾美耳球虫(E.cylindeica)、邱氏艾美耳球虫(E.zuernii)、阿拉巴艾美耳球虫(E.alabamensis)、奥博艾美耳球虫(E.auburnensis)、巴西艾美耳球虫(E.brasiliensi)、拔克朗艾美耳球虫(E.bukidnonensis)、加拿大艾美耳球虫(E.canadensis)、椭圆艾美耳球虫(E.ellipsoidalli)、皮利他艾美耳球虫(E.pellita)、亚球形艾美耳球虫(E.subspherical)和怀俄明艾美耳球虫(E.wyomingensis);所有牛场都存在混合感染的情况；在14个牛场中检测出平均每克粪便中球虫卵囊数(OPG)最高为5 776.32,最低为28...     

江苏省泰州市猪球虫种类的初步调查

本文报道了江苏省泰州市司巷乡猪球虫的感染种类以及不同年龄、不同临床症状的猪球虫感染率，共检查40头猪，猪球虫阳性22头，阳性率为55％。共查到1科2属9种球虫卵囊，其中艾美耳属（Eimeria）有8种，即猪艾美耳球虫（E.suis）、蒂氏艾美耳球虫（E.debliecki）、光滑艾美耳球虫（E.cerdonis）、粗糙艾美耳球虫（E.scabra）、新蒂氏艾症状耳球虫（E.neodebliecki）、豚艾美耳球虫（E.porci）、最小艾美耳球虫（E.perminuta）和有刺艾美耳球虫（E.spinosa）；等孢属（Isospora）1种，即猪等孢球虫（I.suis）。     

安徽省部分散养鸡场球虫感染情况的调查

为了解安徽省散养鸡群球虫感染及虫种分布情况,从安徽省多地散养鸡场采集鸡新鲜粪便829份,分别采用基于7种鸡球虫内转录间隔区1(ITS-1)基因的套式PCR进行检测,并对获得的样本进行测序和分析,以验证球虫虫种。结果显示,调查的所有散养鸡场均有球虫感染,堆型艾美耳球虫(E.acervulina)、巨型艾美耳球虫(E.maxima)和和缓艾美耳球虫(E.mitis)是调查最常见的虫种。混合感染很普遍,以E.acervulina+E.maxima,E.acervulina+E.mitis,E.maxima+E.mitis及E.acervulina+E.maxima+E.mitis混合感染较为常见。调查结果显示,安徽省散养鸡群中球虫感染率较高,应加强对散养鸡场球虫病的综合防控。     

鸡球虫病的诊断和治疗

鸡球虫病是养禽业常见的一种疾病，是由艾美耳科的8种艾美耳球虫引起的。其中柔嫩艾美耳球虫、毒害艾美耳球虫、堆型艾美耳球虫和巨型艾美耳球虫的致病性最为严重。柔嫩艾美耳球虫寄生于盲肠，故称盲肠球虫病；其余7种球虫寄生于小肠，因而称为小肠球虫病。病鸡表现贫血、消瘦和血痢，急性感染可造成大批死亡；中轻度感染主要影响生长发育，并降低对其它疾病的抵抗力。     

杨凌某山羊场球虫种类及感染情况的调查研究

为了解陕西杨凌某奶山羊场山羊球虫的感染状况,采用粪便漂浮法、斯陶尔法和卵囊培养法等对山羊的球虫感染情况进行了初步调查,并对各虫种进行鉴定。结果检获12种艾美耳球虫(Eimeria),即艾丽艾美耳球虫(E.alijevi)、小型艾美耳球虫(E.parva)、浮氏艾美耳球虫(E.faurei)、阿氏艾美耳球虫(E.arloingi)、颗粒艾美耳球虫(E.granulosa)、山羊艾美耳球虫(E.caprina)、羊艾美耳球虫(E.caprovina)、槌状艾美耳球虫(E.crandallis)、克氏艾美耳球虫(E.christenseni)、阿普艾美耳球虫(E.apsheronica)、阿撒他艾美耳球虫(E.ahsata)和错乱艾美耳球虫(E.intricata)。山羊球虫平均感染率为95.2%(40/42),羔羊感染率为100%。平均感染强度(每克粪便卵囊数)为1 086OPG,多数羊为2种~6种卵囊混合感染。优势种为阿氏艾美耳球虫(E.arloingi)、小型艾美耳球虫(E.parva)、艾丽艾美耳球虫(E.alijevi)、山羊艾美耳球虫(E.caprina)、阿撒他艾美耳球虫(E.ahsata)和错乱艾美耳球虫(E.intricata)。     

随机扩增多态性DNA技术对鸡的3种艾美耳球虫的鉴别

应用随机扩增多态性DNA技术（RAPD）对寄生于鸡的3种艾美耳球虫：布氏艾美耳球虫、堆型艾美耳球虫、柔嫩艾美耳球虫的基因组DNA进行分析,发现大多数引物对不同种球虫的DNA扩增产物的电泳带存在明显差异,证明RAPD技术可用于同宿主艾美耳球虫虫种的鉴定。     

Eimeria brunetti and Eimeria necatrix in chickens of Argentina and confirmation of seven species of Eimeria

Ten poultry farms (broiler breeder pullets, layer pullets, and broilers) in the provinces of Entre Rios and Buenos Aires in Argentina were examined for presence of Eimeria spp. Litter samples obtained from flocks 7-11 wk old were taken to the laboratory for oocyst counting and sporulation, then concentrated for inoculation into coccidia-free chickens. Species were identified by prepatent period, oocyst size, location and appearance of lesions in the intestine, microscopic examination of mucosal smears, and histology (to confirm Eimeria brunetti). On this basis, Eimeria praecox was found in two samples, Eimeria mitis in two, Eimeria acervulina in nine, Eimeria maxima in seven, Eimeria necatrix in three, Eimeria tenella in seven, and E. brunetti in four. These results confirm the presence of all seven recognized species of Eimeria in chickens in the Republic of Argentina.     

Tracing the emergence of drug-resistance in coccidia (Eimeria spp.) of commercial broiler flocks medicated with decoquinate for the first time in the United Kingdom

Decoquinate is a quinolone coccidiostat introduced during 1967 as an in-feed prophylactic for broiler chickens. Despite early drug-resistance problems and its age, the drug is still used commercially worldwide. Decoquinate here serves as a valuable model in a field study that addresses the dynamics and economic impact of the development of coccidial resistance to potent synthetic anticoccidial drugs. The results of this unique, hitherto unpublished, study on the initial emergence of resistance of avian coccidia (Eimeria spp.) to a new drug in the field may be of strategic value in the continued use of decoquinate or the introduction of new drugs. The commercial performance of the first 3-5 crops of broilers to be medicated with decoquinate on each of six farms was monitored during 14 months in 1968-1969, supplemented by assessments of the species, population dynamics and decoquinate-resistance of coccidia isolated from each farm. During the rearing of each flock in a single shed on each farm, oocysts were counted in fresh faecal samples collected on three occasions, and the species were identified by their morphology if possible, supported if necessary by the biological characteristics of infections in chickens. E. acervulina was the most common species, followed by E. mitis, E. maxima, E. tenella and E. praecox. E. brunetti occurred rarely, and E. necatrix was not found. Decoquinate-resistance was evident in several species during the rearing of the first decoquinate-medicated crop on each farm, although clinical coccidiosis did not occur. It was concluded that inherently resistant mutants of E. acervulina, E. brunetti, E. maxima, E. tenella, and probably also E. mitis and E. praecox, were selected from field populations by 6 weeks during their first exposure to decoquinate. During up to four more subsequent crops, cycling of resistant parasites stimulated host immunity, which had no obvious adverse impact on commercial performance. There was no apparent seasonal effect. A hypothesis is proposed to explain the sudden and rapid emergence of quinolone-resistance in the coccidia, and why bird health was not thereby compromised in these circumstances.     

Quantitative real-time PCR assays for detection and quantification of all seven Eimeria species that infect the chicken

The development and validation of real-time quantitative PCR (qPCR) assays specific to all seven Eimeria species that cause coccidiosis in the chicken is described. The presented work utilizes previously published assays for Eimeria maxima, E. necatrix and E. tenella and adds assays for E. acervulina, E. brunetti, E. mitis and E. praecox. These assays target unique single copy sequences derived from sequence characterized amplified region (SCAR) markers. All seven qPCR markers were sequenced from multiple strains and confirmed to be non-polymorphic and identical to the original SCAR sequence. Sequences conserved within each species were chosen with the aim of developing genuinely universal markers, providing global coverage. An exact match for the primers and TaqMan(?) probe during PCR cycling enables precise relative quantification of multiple species in a mixture regardless of the strains present. All markers utilized in these qPCR assays are absolutely species-specific and support reproducible quantification across a wide linear range, unaffected by the presence of non-target species or other contaminating DNA. The sensitivity of these assays indicates that DNA equivalent to a single sporulated oocyst can be consistently detected. These assays will be a valuable tool from both industry and research perspectives. Comparison of our panel of qPCR assays with results derived by microscopy, the traditional Gold Standard, using poultry farm field samples support their efficacy.     

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.     

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.     

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.     

Isolation and pathogenicity of Australian strains of Eimeria praecox and Eimeria mitis

Objective To determine the presence of E praecox and E mitis in Australia, to isolate representative strains of these species from chickens and determine their pathogenicity.
Design Morphological, physiological and cross protection studies were undertaken to confirm the identity of Australian isolates of E praecox and E mitis.
Procedure Oocysts were isolated from a backyard flock at Jimboomba, southeastern Queensland and numbers of E praecox and E mitis enriched by passage in chickens immune to five other species of poultry Eimeria . Oocysts of mean conformation and size of the two species were purified by single oocyst passage. Two isolates that closely matched recorded parameters for E praecox and E mitis were selected and designated JP and JM respectively. The cross protection between the isolates and E acervulina was determined by infection and challenge experiments. The virulence of the two isolates was determined by comparing weight gains of groups of birds inoculated with JP isolate or JM isolate with untreated groups.
Results Isolates JP and JM most closely matched recorded parameters of E praecox and E mitis respectively. Groups of chickens, previously infected with JP and JM isolates, showed no significant protection against infection with E acervulina . In a separate trial, groups of susceptible chickens inoculated with 10⁵ oocysts of JP and JM isolates showed significantly reduced weight gains compared with untreated controls.
Conclusion Isolates JP and JM are E praecox and E mitis respectively, confirming the presence of these species in Australia. These isolates were found capable of causing significant reductions in weight gains in susceptible chickens.     

HACCP体系对肉鸡生产中沙门氏菌的控制效果

在上海某大型鸡场所辖的5 个肉鸡场严格按照肉鸡生产SSOP程序基础之上制定并实施HACCP 程序,为了验证HACCP体系的应用效果,我们以出口禽肉中要求最严格的病原之一沙门氏菌为例,比较鸡场在应用HACCP程序之前和之后的沙门氏菌感染情况。2003年3月份和9 月份在5 个实施HACCP的鸡场和5 个相同规模的对照鸡场采集泄殖腔拭子,每个鸡场480 份样品,检测沙门氏菌的感染率。结果实施HACCP的鸡场沙门氏菌阳性率由实施HACCP 前的1.17%(28/2 400)下降到0.04%(1/2 400),而同期5个对照鸡场沙门氏菌阳性率分别为1.25%(30/2 400)、1.67% (40/2 400)。由此可见,HACCP程序的实施,对鸡场沙门氏菌的控制有明显的效果。     

A survey of sensitivity to anticoccidial drugs in 60 isolates of coccidia from broiler chickens in Brazil and Argentina

Coccidia were isolated from 90 broiler farms in 15 poultry-producing areas in Brazil and Argentina. Sixty isolates were tested for sensitivity to 7 anticoccidial drugs. The common species were: a) Eimeria tenella, 47 isolates; b) E. maxima, 49 isolates; c) E. acervulina, 44 isolates; d) E. mitis, 26 isolates; and e) E. brunetti, 12 isolates. Isolates were considered sensitive to drugs if intestinal lesion scores of medicated broilers were reduced by at least 50% compared with unmedicated infected broilers or if weight gain was at least 75% of that of uninfected birds in a 6-day laboratory test. According to lesion scores, there was evidence of resistance or seriously reduced sensitivity to monensin in 20 isolates, narasin in 29, salinomycin in 11, maduramicin in 1, clopidol in 36, amprolium in 40, and nicarbazin in 1. According to broiler weight gain, there was resistance to monensin in 36 isolates, narasin in 32, salinomycin in 28, maduramicin in 2, clopidol in 28, amprolium in 50, and nicarbazin in 4. These results suggested incomplete cross resistance of coccidia to polyether ionophorous drugs. The degree of resistance might be explained by previous patterns of use of these drugs.     

A preliminary study of Salmonella, verocytotoxigenic Escherichia coli/Escherichia coli O157 and Campylobacter on four mixed farms

The aims of this study were to investigate the incidence of Salmonella, verocytotoxigenic Escherichia coli (VTEC)/Escherichia coli O157 and Campylobacter on four mixed farms and to characterize the isolates in terms of a range of virulence factors. Eighty-nine composite (five different samples from the same animal species combined) faecal [cattle (24), pigs (14), sheep (4), poultry (4), horses (7), deer (4), dogs (9), rodents (2) and wild birds (20)] samples, 16 composite soil samples plus 35 individual water samples were screened using culture-based, immunomagnetic separation and molecular methods. Salmonella was detected in bovine faeces, cattle and poultry house water. Salmonella serotypes/phage types included Dublin, Kiel and Typhimurium DT193, and most isolates were spvC, invA and rck positive. The pefA and rck genes were found exclusively in the non-Typhimurium strains, while Salmonella Dublin and Salmonella Kiel strains carried Salmonella genomic island I marker(s). VTEC/E. coli O157 were found in deer and dog faeces only. The E. coli O157 isolate was an enteroinvasive E. coli, while the VTEC isolate was untypable but carried the vt1, eaeA, hlyA, tir and eptD genes. This article reports the first confirmed carriage of E. coli O157 in Irish deer. Campylobacter species were not detected over the course of this study. It was concluded that [1] Salmonella, VTEC and Campylobacter have low (<5%) prevalence or are absent on the farms in this study; [2] water was an important source of bacterial pathogens; [3] both dogs and deer may act as a source of pathogenic E. coli and [4] key virulence and resistance determinants are widespread in farm Salmonella strains. This study highlights the need to control water as a source of pathogens and suggests that the domestic pets and deer should be considered in any farm risk assessment.