艾美耳球虫是鸡球虫病的病原体，采用疫苗防治是当前研究的热点之一。其表面抗原是一种有效的疫苗候选分子，本文对卡介苗（BCG）、乳酸乳球菌（LL）、植物乳杆菌（LP）、鼠伤寒沙门氏菌（St）、枯草芽孢杆菌（Bs）、粪肠球菌（Efs）、大肠埃希菌（Ec）、蓝细菌（CB）、酵母、蜥蜴利什曼原虫（Lt）、鸡痘病毒（FPV）、火鸡疱疹病毒（HVT）、牛痘病毒（VV）和昆虫杆状病毒等载体介导的艾美耳球虫表面抗原疫苗的研制现状做一概述，以期为新型疫苗的开发和应用提供参考依据。

寄生虫病仍然是全球最大的健康问题，给贫困地区造成巨大的经济负担。目前用于治疗原虫病和蠕虫病的药物存在一定缺陷，亟待研发更为有效的治疗药物。小檗碱最早从黄连的根茎中提取获得，是一类重要的抗炎药物。小檗碱的衍生物通过修饰小檗碱的结构位点获得，除了具有抗菌和抗微生物活性以外，小檗碱及其衍生物还具有显著的抗寄生虫活性。本文概括了小檗碱及其衍生物抗原虫（利什曼原虫、锥虫、刚地弓形虫、恶性疟原虫和柔嫩艾美尔球虫）和蠕虫（曼氏血吸虫、日本血吸虫、细粒棘球绦虫和犬弓首蛔虫）感染的作用，以期为相关研究工作者提供参考。

目的 了解福建省实验兔球虫的感染情况.方法 采集福建省6个地级市2家实验兔生产企业和12家使用单位的实验兔粪样,通过饱和盐水漂浮法对卵囊进行收集,测定每克粪便中卵囊数(OPG)并对孢子化的卵囊进行鉴定.结果 本次调查共收集和分析了195个样本,阳性样本28个,总感染率为14.36％,OPG平均值为2 333.33.2个实验兔生产企业感染率为22.22％,12家实验兔使用单位感染率为13.10％.共检出10种兔球虫,均为混合感染,以斯氏艾美耳球虫为优势虫种.普通级实验兔的球虫感染率为15.30％,清洁级实验兔未见球虫感染.幼兔球虫检出率最高,感染率为17.50％,青年兔次之,成年兔感染率最低,感染强度最高.我省生物制药企业实验兔球虫感染强度最大,OPG平均值为5 678.39,感染率也较高.在医院、高校等科研单位,实验兔也存在不同程度的球虫感染.结论 福建省实验兔球虫感染率较低.但日常工作中还是应进一步加强实验兔球虫病的监测与防治,并建议将兔球虫病纳入国家标准中普通级实验兔必要时的检测项目.

目的 建立荧光定量PCR检测兔斯氏艾美耳球虫的方法.方法 根据斯氏艾美耳球虫(内转录间隔区1)ITS1序列区设计特异性引物,构建重组质粒,并作为标准品绘制标准曲线,对建立的荧光定量PCR检测方法进行特异性、敏感性和重复性试验.结果 建立的荧光定量PCR能特异性地检测兔斯氏艾美耳球虫,且可检测到含一个卵囊DNA的样本.该方法的重复性较好,组内、组间重复试验的变异系数均小于2％.结论 建立了一种特异性好、敏感度高、可靠的检测兔斯氏艾美耳球虫荧光定量PCR方法.

目的 研究柔嫩艾美耳球虫顶膜抗原-1(EtAMA1)与微线蛋白-2(EtMIC2)之间的相互作用. 方法 以柔嫩艾美耳球虫子孢子cDNA为模板,PCR扩增EtAMA1和EtMIC2基因,构建酵母双杂交诱饵载体pGBKT7-EtAMA1及捕获载体pGADT7-EtMIC2并转化Y2HGold酵母菌中,观察酵母在营养缺陷选择培养基上的生长情况.原核表达 GST-EtAMA1和His-EtMIC2融合蛋白,纯化后进行GST-Pulldown试验.同时构建真核表达载体pcDNA3.1-His- EtAMA1和pcDNA3.1-HA-EtMIC2并转染HEK-293T细胞,收集细胞裂解液,进行免疫共沉淀试验.在此基础上构建双分子荧光互补载体pEtAMA1-Myc-LC151和pEtMIC2-HA-KN151并转染HEK-293T细胞,激光共聚焦观察转染细胞荧光发生情况. 结果 诱饵基因和捕获基因均对酵母双杂交系统无自激活和毒性作用,pGBKT7-EtAMA1/ pGADT7-EtMIC2共转化菌能激活酵母双杂交报告系统;原核表达并纯化GST-EtAMA1和His-EtMIC2融合蛋白进行 GST-Pulldown试验,二者可在体外相互作用;EtAMA1和EtMIC2可在HEK-293T细胞中共表达,免疫共沉淀试验进一步证实二者间的相互作用;双分子荧光互补技术验证EtAMA1和EtMIC2在细胞内存在相互作用. 结论 通过酵母双杂交、GST-Pulldown、免疫共沉淀及双分子荧光互补技术验证柔嫩艾美耳球虫顶膜抗-1(EtAMA1)与微线蛋白-2(Et- MIC2)之间存在相互作用,这为揭示微线蛋白在鸡球虫入侵宿主细胞过程中的分子机制提供了理论基础.

Eimeria stiedae (E.stiedae),the most common protozoan pathogen of rabbits,causes coccidiosis.This disease is problematic for rabbit breeders [1].There is little information about the cellular and molecular biology of this pathogen and a lack of genetic tools.Genetic manipulation is necessary to advance our understanding of parasite biology and ultimately develop vaccines and treat diseases.DNA-mediated and viral RNA-mediated transfection systems have successfully been established in many protozoan parasites [2,3].It was discovered that E.stiedae possesses a dsRNA virus (EsRV1),and more recently,the genome of this virus has been sequenced.To date,however,there is no report on the use of viral RNA-mediated transfection for E.stiedae.In this study,we report a transient viral RNA-mediated transfection of E.stiedae using enhanced green fluorescent protein (EGFP) as a reporter gene.Our study provides a valuable genetic tool for studying the biology of E.stiedae.

Eimeria acervulina severely damages the intestinal mucosa in infected poultry, resulting in major economic losses [1].The study of the interaction between Eimeria and host cells is the theoretical basis for the prevention and treatment of coccidiosis [2].However, the specific mechanism of E.acervulina invasion is not very clear, so the interaction between the invading protein and the host awaits extensive study.

The production and structure of animal signals may depend on an individual's health status and may provide more than one type of information to receivers.While alarm calls are not typically viewed as health condition dependent,recent studies have suggested that their structure,and pos-sibly their propensity to be emitted,depends on an individual's health condition and state.We asked whether the propensity of yellow-bellied marmots(Marmota flaviventer)to emit calls is influ-enced by their immunological or parasite status,by quantifying both trap-elicited and natural call-ing rates as a function of their neutrophil-to-lymphocyte(NL)ratio,the presence of a blood borne trypanosome,and the presence of several intestinal parasites(Eimeria sp.,Entamoeba sp.,and Ascaris sp.).We fitted mixed effects models to determine if the health measures we collected were associated with the probability of calling in a trap and with annual rates of natural alarm calling.Marmots infected with a blood-borne trypanosome were marginally more likely to call naturally and when trapped,while those infected with the intestinal parasite Ascaris were less likely to call when trapped.NL ratio was not directly associated with in-trap calling probability,but males were more likely to call when they had higher NL ratios.Thus,health conditions,such as parasite infec-tion and immune system activation,can modulate the production of alarm signals and potentially provide information to both predators and prey about the caller's condition.Playback experiments are required to confirm if receivers use such information.

目的 建立兔斯氏艾美耳球虫(Eimeria stiedai,E.stiedai)感染模型和巢式PCR诊断方法.方法 利用剖检、显微镜观察、血液生化和病理组织学检查等方法对建立的兔E.stiedai模型进行鉴定.通过收集卵囊,提取E.stiedai的DNA,设计特异性引物,建立E.stiedai巢式PCR检测方法.结果 临床剖检:兔腹围明显增大,解剖见肝脏肿大,肝脏表面和实质布满白色及淡黄色结节,胆囊和胆管肿大,胆汁呈淡黄色.显微镜观察:虫卵大小为(31.72～38.43)μm x(18.10～22.69)μm.血液生化检查:球蛋白(GLOB)指标偏高,肌酐(CREA)和碱性磷酸酶(ALKP)指标偏低,其余检测指标均在参考值范围内.病理组织学检查:肝组织和胆管中见大量粉红色E.stiedai虫卵.巢式PCR检查:最低检测限为1个卵囊DNA样本和1.14 × 103拷贝数质粒,阴性对照和空白对照均未出现条带,重复性实验变异性系数＜ 5％.结论 成功构建兔E.stiedai感染模型,建立的巢式PCR诊断方法可扩增出E.stiedai的特异片段,敏感性强,重复性好.

Short-term elevation of glucocorticoids(GCs)is one of the major physiological mechanisms by which vertebrates cope with challenging environmental or social factors(stressors).However,when exposure to stressors occurs repeatedly or over a prolonged period of time,animals may ex-perience chronic elevation of GCs,which reduces the immune response efficiency and can lead to higher intensity of parasitic infection.Here,we used invasive gray squirrels Sciurus carolinensis introduced in Northern Italy and their 2 most prevalent gastrointestinal parasites,the nematode Strongyloides robustus and coccidia of the genus Eimeria,as a model to investigate relationships among macroparasite infection and concentrations of fecal glucocorticoid metabolites(FGMs),an integrated measure of circulating GCs.Our results revealed an association of FGMs with infection by St.robustus,but not with coccidia.Individuals with higher FGMs appear to be responsible for the greatest St.robustus egg shedding within gray squirrel populations,thus possibly acting as superspreaders.However,FGMs were negatively associated with adult St.robustus,suggesting that the abundance of adults of this nematode species does not induce elevation in FGMs,but is only affected by it through immune-mediated effects on its fecundity.Finally,the relationship be-tween St.robustus(both eggs and adult parasites)and FGMs was not linear,suggesting that only high levels of physiological stress influence parasite infection.Our findings highlight that the direc-tion and magnitude of the stress-infection relationship may depend not only on the specific host-parasite system,but also on the different life stages of the same parasite.