绵羊艾美虫一新种：（真球虫目：艾美亚目：艾美科）

本文记述了真球虫目、艾美科、固原艾美虫一新种Eimeria guyuanensis sp.nov.的形态。在宿主——绵羊肠道里的寄生部位及体外发育情况。本种与小型艾美虫E.parva的区别在于有卵膜孔和浅极帽。新种与槌形艾美虫E.crandallis的区别在于卵囊较小,孢子囊长宽比例较大,有斯氏体,以及卵囊的寄生部位与体外发育等特征不同。     

柔嫩艾美耳球虫安徽肥西(AHFX)株的分离与鉴定

目的 分离并鉴定安徽肥西病鸡盲肠内的柔嫩艾美耳球虫.方法 通过雏鸡进行单卵囊接种与增殖试验,对所获卵囊用形态学方法进行初步鉴定;运用PCR方法扩增该分离株的ITS-1基因序列,并与相关序列进行比对、构建系统进化树.结果 该球虫分离株孢子化卵囊的平均大小为21.1 μm ×18.0 μm,卵形指数为1.17,潜隐期为140 h;其ITS-1基因序列与柔嫩艾美耳球虫(Eimeria tenella)GQ856310相似性达98.2％,二者的亲缘关系非常接近.结论 该球虫分离株为柔嫩艾美耳球虫(E.tenella),暂命名为柔嫩艾美耳球虫安徽肥西(AHFX)株.     

柔嫩艾美耳球虫孢子发育阶段虫体抑制性消减文库的构建

为了分离鉴定柔嫩艾美耳球虫(Eimeria tenella)孢子发育阶段虫体的差异表达基因,分别以柔嫩艾美耳球虫未孢子化卵囊和孢子化卵囊为驱动组、子孢子为实验组,或未孢子化卵囊为驱动组、孢子化卵囊为实验组,利用抑制性消减杂交(SSH)技术,构建了2个子孢子cDNA消减文库和1个孢子化卵囊cDNA消减文库。随机从3个cDNA消减文库中分别挑取50个克隆,经PCR鉴定2个子孢子cDNA消减文库的重组率都为96%,孢子化卵囊cDNA消减文库的重组率为98%。从每个文库中随机挑取50个克隆测序,并进行同源性比较分析,结果显示:从孢子化卵囊cDNA消减文库中获得了13个单一有效序列,其中8个EST与已知蛋白同源性很高;从2个子孢子cDNA消减文库中共获得了40个单一有效序列,其中9个EST与已知蛋白同源,其余可能为柔嫩艾美耳球虫的新基因。这些结果为分离柔嫩艾美耳球虫新功能基因和进一步探索防治球虫病的方法提供了理论基础。     

柔嫩艾美耳球虫孢子发育阶段虫体抑制性消减文库的构建

为了分离鉴定柔嫩艾美耳球虫（Eimeria tenella）孢子发育阶段虫体的差异表达基因,分别以柔嫩艾美耳球虫未孢子化卵囊和孢子化卵囊为驱动组、子孢子为实验组,或未孢子化卵囊为驱动组、孢子化卵囊为实验组,利用抑制性消减杂交（SSH）技术,构建了2个子孢子cDNA消减文库和1个孢子化卵囊cDNA消减文库。随机从3个cDNA消减文库中分别挑取50个克隆,经PCR鉴定2个子孢子cDNA消减文库的重组率都为96%,孢子化卵囊cDNA消减文库的重组率为98%。从每个文库中随机挑取50个克隆测序,并进行同源性比较分析,结果显示：从孢子化卵囊cDNA消减文库中获得了13个单一有效序列,其中8个EST与已知蛋白同源性很高;从2个子孢子cDNA消减文库中共获得了40个单一有效序列,其中9个EST与已知蛋白同源,其余可能为柔嫩艾美耳球虫的新基因。这些结果为分离柔嫩艾美耳球虫新功能基因和进一步探索防治球虫病的方法提供了理论基础。     

巨型艾美耳球虫孢子化卵囊cDNA文库的构建

为构建巨型艾美耳球虫(Eimeria maxima)孢子倾卵囊cDNA表灰文库,从E.maxima孢子化卵囊中提取总RNA,以总RNA为模板、λTriplex2TM为栽体,利用SMARTTM cDNA文库构建试剂盒构建全长cDNA表达文库.经测定构建的E.maxima cDNA表达文库的原始文库容量为1×106 pfu/ml,扩增后的文库滴度为5×1010pfu/ml,重组率为95%,插入片断主要集中在0.5～1kb之间.根据已知序列设计引物,能从文库中扩增出编码E.maxima免疫球蛋白重链结合蛋白的基因序列片段.结果 表明所构建的E.maxima cDNA表达文库质量良好,为克隆、筛选E.maxima的功能性基因奠定了基础.     

柔嫩艾美耳球虫脱囊过程的扫描电镜观察

本文对柔嫩艾美耳球虫的体外脱囊过程进行了扫描电镜观察。结果表明该虫表面粗糙,无其他生物学结构。孢子囊呈橄榄状,斯氏体明显。在脱囊介质作用下,孢子囊壁强度发生变化,出现不同程度的塌陷和皱折。未观察到因斯氏体消失而直接留下的开口。子孢子香蕉状,表面有数个微孔呈不规则排列,顶器短突状,无其他表面结构。     

大足鼠耳蝠的艾美球虫一新种

1980年10月,我们在昆明市花红洞(岩洞)采到大足鼠耳蝠Myotis ricketti Thomas151只,其中105只感染一种艾美球虫,经鉴定为一新种,现描述如下。标本保存于云南大学生物系动物学教研室。昆明艾美球虫Eimeria kunmingensis,新种(图1) 卵囊近圆球形或为宽椭圆形。卵囊壁光滑,由二层组成,外层较厚,约1.0微米,淡黄     

高原鼠兔寄生艾美耳球虫(顶复器门,艾美耳科)二中国新纪录种

记述从高原鼠兔Ochotona curzoniae新鲜粪便收集的艾美耳属Eimeria球虫二中国新纪录种:E.worleyi Lepp,Todd & Samuel,1972和E.klondikensis Hobbs & Samuel,1974,高原鼠兔是其寄生新宿主.鼠兔标本保存于青藏高原生物标本馆,孢子化卵囊标本保存于中国科学院西北高原生物研究所高原生物适应与进化重点实验室.     

柔嫩艾美耳球虫HSP基因的克隆、表达及鉴定

为研究柔嫩艾美耳球虫热激蛋白(Heat shock proteins,HSPs)的生物学特性,应用RACE和RT-PCR技术,从柔嫩艾美耳球虫子孢子中首次克隆获得了EtHSP的全长cDNA(GenBank Accession No.FJ911605)。EtHSP包含一个1455 bp的开放阅读框,编码484个氨基酸,预测表达蛋白的分子量大小为53.5 kD。应用Real-time PCR对柔嫩艾美耳球虫不同发育阶段(未孢子化卵囊、孢子化卵囊、子孢子和裂殖子)表达量进行分析,发现该基因在子孢子阶段的表达明显高于其他阶段。同时,构建了原核表达重组质粒pET28a(+)-EtHSP,转化到大肠杆菌BL21(DE3)中,经IPTG诱导表达后,对表达产物进行SDS-PAGE及Western blotting分析。结果显示,重组质粒pET28a(+)-EtHSP在大肠杆菌中以包涵体形式表达,经1 mmol/L IPTG诱导6 h后的表达量最高,该蛋白可被抗柔嫩艾美耳球虫的多克隆抗血清识别,表明该蛋白具有较好的反应原性。本研究结果为进一步研究该基因的生物学功能奠定了基础。     

肠艾美耳球虫孢子发育与裂殖生殖研究

用单卵囊分离技术纯化肠艾美耳球虫Eimeria intestinalis Cheissin,1948卵囊。卵囊呈宽梨形,平均大小为27.38×19.97μm。在室温21—26℃,第63和116小时完成孢子化率分别为31％和71％。人工感染无球虫兔,潜隐期9天,排卵囊的持续期为9天,高峰期为感染后第12—13天。裂殖生殖4代,第一代裂殖体寄生于空肠和回肠的肠腺上皮细胞内,出现时间为感染后61—85小时;裂殖体有大小两种类型。第2代至第4代裂殖体寄生于空肠、回肠,寄生部位扩展至肠绒毛上皮细胞,有大、中、小三种类型裂殖体。第2至第4代分别出现于感染后96—132小时;144—180小时;192—240小时。感染后73—216小时之间均见有含两个裂殖子的小裂殖体,大小为2.5—5.9×3—9.48μm;感染后96—240小时,见粗细两种类型的裂殖子,粗裂殖子有核1—8个,细裂殖子多为一个核。裂殖生殖寄生于空肠、回肠;仅在216小时曾在蚓突的个别绒毛和腺上皮细胞内见有裂殖体。12指肠、盲肠和结肠均未见虫体寄生。作者特别瞩目于试验中发现的裂殖体和裂殖子的多型现象,并将无性世代划分为4代。     

A new species of Eimeria (Apicomplexa: Eimeriidae) from the eastern pipistrelle, Perimyotis subflavus (Chiroptera: Vespertilionidae), in Arkansas

During November 2009 and March 2010, 20 adult eastern pipistrelles, Perimyotis (= Pipistrellus) subflavus, were collected from Polk County, Arkansas, and their feces were examined for coccidian parasites. Two (10%) of the bats were found to be passing oocysts of an undescribed species of Eimeria. Oocysts of Eimeria heidti n. sp. were ovoidal to ellipsoidal, 26.1 × 20.5 μm (23-31 × 18-23 μm), with a bilayered wall, externally rough, internally smooth, and with a shape index of 1.3. Micropyle and oocyst residuum were absent, but a subspherical polar granule was often present. Sporocysts were ovoidal, 13.0 × 8.8 μm (11-15 × 7-13 μm), the shape index was 1.6, a Stieda body was present and sub-Stieda and para-Stieda bodies were absent. A sporocyst residuum consisting of multiple globules dispersed along the perimeter of the sporocyst and between the sporozoites were present, sporozoites were elongate, with a subspherical anterior refractile body and elongate posterior refractile body; a nucleus not discernible. This is the second coccidian reported from this host and the fourth instance of a coccidian species reported from an Arkansas bat.     

A new eimeriid (Apicomplexa) species from endangered Attwater's prairie chickens (Tympanuchus cupido attwateri) in Texas

The Attwater's prairie chicken (APC; Tympanuchus cupido attwateri Bendire, 1894) has been a federally listed endangered species since 1967. Several captive propagation programs consisting of small populations are being used to keep this species from extinction. Fecal samples were collected from APCs in April 2007 and again in August 2008 from 2 separate captive propagation facilities in Texas after clinical signs of coccidiosis were observed. One Eimeria species was observed (Eimeria attwateri), which we describe as a putative new species. Sporulated oocysts are ellipsoidal, 30.0 × 18.4 (27.4-31.3 × 16.0-22.4) μm. Oocysts have a smooth wall 0.7 μm thick and lack both a micropyle and oocyst residuum, but 1 ellipsoidal polar granule is present, 2.3 × 1.9 (2.1-2.4 × 1.7-2.0) μm. Sporocysts have a nipple-like Stieda body with a rounded opposite end and are 14.0 × 7.1 (10.2-16.8 × 6.0-9.2) μm. The sporocysts contain a sporocyst residuum usually consisting of 2-4 dispersed globules, and each sporozoite contains 2 large posterior spheroid refractile bodies 3.4 μm wide. Nucleotide sequence amplified from the 18S rDNA does not match any DNA sequence information for publicly available Eimeria species, and phylogenetic reconstructions place this species with other eimerians from Galliformes. The discovery of a potentially pathogenic species of Eimeria in captive APCs is of great importance, and managers should be aware of the potential devastating effect(s) this parasite could have on the APC conservation programs.     

Two new Eimeria species (Apicomplexa: Eimeriidae) from the yellow-crowned Amazon Amazona ochrocephala (Aves: Psittacidae) in Brazil

In this study, we describe 2 new species of Eimeria associated with the yellow-crowned Amazon Amazona ochrocephala. Eimeria amazonae n. sp. has bilayered, ellipsoidal, and smooth oocysts that measure 48.9 × 36.2 μm; the length/width ratio is 1.35. The micropyle and oocyst residuum are both absent, but the polar granule is present. Ovoidal sporocysts are 22.2 × 11.9 μm. Stieda and sub-Stieda bodies and sporocyst residuum are present. The 2 elongate sporozoites are curved and measure 18.1 × 3.4 μm; both have 2 refractile bodies. Eimeria ochrocephalae n. sp. has bilayered, ellipsoidal, and smooth oocysts that measure 43.8 × 27.7 μm; the length/width ratio is 1.58. The micropyle and oocyst residuum are absent, but the polar granule is present; ovoidal sporocysts are 20.6 × 10.1 μm. Stieda and sub-Stieda bodies and sporocyst residuum are present; 2 elongate and curved sporozoites are 15.8 × 3.4 μm, each of which has 2 refractile bodies.     

Coccidian Parasites (Apicomplexa: Eimeriidae) from Insectivores: New Species from Shrew Moles (Talpidae) in the United States1

All of 18 shrew moles, Neurotrichus gibbsii, collected in Oregon and Washington were infected with one or more species of coccidia. Three eimerians and one isosporan were identified and described as new species. Sporulated oocysts of Eimeria heterocapita n. sp. were subspheroid to ellipsoid, 25.5 times 21.4 (23–27 times 18–23) μm. A membranous, cap-like structure was present at one pole of the oocyst, but a micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts were 13.6 times 10.0 (12–15 times 9–11) μm; a compact sporocyst residuum was present, but Stieda, sub-, and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria neurotrichi n. sp. were avoid, 17.6 times 13.6 (16–20 times 11–16) μm; micropyle and oocyst residuum were absent, but a polar body was present. Ovoid sporocysts were 10.7 times 5.5 (9–12 times 5–6) μm; Stieda body and sporocyst residuum were present, but sub- and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria parastiedica n. sp. were subspheroid, 27.4 times 25.5 (25–30 times 22–28) μm; micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts, pointed at both ends, were 18.3 times 10.4 (16–20 times 9–11) μm; Stieda, sub-, and parastieda bodies were present as was a sporocyst residuum. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Isospora neurotrichi n. sp. were subspheroid, 13.9 times 12.0 (11–16 times 10–15) μm; micropyle and oocyst residuum were absent, but 1–3 polar bodies were present. Ellipsoid sporocysts were 9.2 times 6.1 (8–11 times 5–8) μm; sub- and parastieda bodies were absent, but a Stieda body and sporocyst residuum were present. This species was found in 17 of 18 (94%) hosts. Ten of 18 (56%) hosts were seen to be naturally infected with only one coccidium.     

A new eimerian species (Apicomplexa: Eimeriidae) from the blue-fronted Amazon parrot Amazona aestiva L. (Aves: Psittacidae) in Brazil

The Neotropical psittacine species Amazona aestiva, commonly known as the blue-fronted Amazon, is one of the most common and best-known psittacine birds kept as a pet worldwide. However, very little is known about the diseases or parasites of these birds. In this study, we describe a new species, Eimeria aestivae, associated with these parrots. The new species is characterized by: ovoid smooth oocysts (n = 60), 36.8 (33.2-41.5) × 23.7 (21.7-25.7) μm, length/width ratio = 1.55; polar granule present; ellipsoidal sporocysts (n = 25), 19.8 (17.5-21.6) × 9.3 (8.3-9.9) μm; Stieda, sub-Stieda body, and sporocyst residuum present. Sporozoites (n = 20), 2 per sporocyst, elongate and curved, 17.6 (15.8-19.2) × 3.8 (3.2-4.8) μm; each with 2 refractile bodies. The oocysts of the other 2 eimerian species described for Amazona are larger than those of the presented species, but they all seem to be closely related because of some similarities among them.     

Two new eimerians (Apicomplexa) from insectivorous mammals in Madagascar

Fecal samples from 126 insectivorous mammals in Madagascar were collected between spring 1999 and fall 2001. In the Afrosoricida, 21 species in 5 genera were sampled, including 17 species of Microgale (31/96, 32% infected), Hemicentetes semispinosus (1/2, 50%), Oryzorictes hova (1/5, 20%), Setifer setosus (8/13, 61.5%), and Tenrec ecaudatus (5/8, 62.5%); in the Soricomorpha, only Suncus murinus was examined and 1/2 (50%) were infected. Two morphotypes of eimeriid oocysts, representing 2 presumptive new species, were found in 47 (37%) infected animals; only 2 afrosoricid hosts (2% of all hosts, 4% of infected hosts) had both oocyst morphotypes. Sporulated oocysts of the first morphotype, Eimeria tenrececaudata n. sp., are subspheroidal, 18.8 × 17.4 (17-22 × 15-20), with a length∶width ratio (L/W) of 1.1 (1.0-1.2); they lack a micropyle but may contain 0-2 polar granules and a single, small round oocyst residuum, 3 × 2.3. Sporocysts are lemon-shaped, 9.9 × 6.6 (9-11 × 5-8), with a L/W of 1.5 (1.2-2.0); they have a prominent, slightly flattened Stieda body and a substieda body but lack a parastieda body. The sporocyst residuum consists of only a few granules between the sporozoites, which are sausage-shaped and have a large posterior refractile body. Oocysts of the second morphotype, Eimeria setifersetosa n. sp. are spheroidal to subspheroidal, 30.1 × 28.6 (27-34 × 25-34), with a L/W of 1.1 (1.0-1.2); they lack both micropyle and oocyst residuum, but 1-2 polar granules are usually present. Sporocysts are subspheroidal to broadly ellipsoidal, 9.6 × 7.3 (9-11 × 6-8), with a L/W of 1.3 (1.1-1.7); they have a broad Stieda body, lack sub- and parastieda bodies, and have a residuum of a few granules scattered throughout the sporocyst. Sporozoites were not clearly defined, but what seemed to be a single large refractile body is seen, presumably in each sporozoite.     

Eimeria auratae n. sp. (Apicomplexa: Eimeriidae) infecting the lizard Mabuya aurata in Saudi Arabia

Eimeria auratae n. sp. was described from the gall bladder of the lizard Mabuya aurata collected at Al-Hofuf village, eastern region, Saudi Arabia. Morphology of sporulated as well as non-sporulated oocysts were studied. Sporulated oocysts were ellipsoidal 22-31.5x13.5-21.8 (27.7x18.5) microm with smooth brownish-yellow bilayered wall, 1.1 (0.9-1.3) microm. Micropyle, polar granule and oocyst residuum were absent. Sporocysts were ellipsoidal 10.5-12.8x7.5-9 (11.8x8.5) microm. Sporocyst residuum was present but Stieda body was absent. Sporozoites were crescent-shaped, blunt at one end and slightly tapered at the other. Eimeria species from Scincidae were compared.     

Coccidian parasites (Apicomplexa: Eimeriidae) from insectivores: new species from shrew moles (Talpidae) in the United States

All of 18 shrew moles, Neurotrichus gibbsii, collected in Oregon and Washington were infected with one or more species of coccidia. Three eimerians and one isosporan were identified and described as new species. Sporulated oocysts of Eimeria heterocapita n. sp. were subspheroid to ellipsoid, 25.5 X 21.4 (23-27 X 18-23) micron. A membranous, cap-like structure was present at one pole of the oocyst, but a micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts were 13.6 X 10.0 (12-15 X 9-11) micron; a compact sporocyst residuum was present, but Stieda, sub-, and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria neurotrichi n. sp. were ovoid, 17.6 X 13.6 (16-20 X 11-16) micron; micropyle and oocyst residuum were absent, but a polar body was present. Ovoid sporocysts were 10.7 X 5.5 (9-12 X 5-6) micron; Stieda body and sporocyst residuum were present, but sub- and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria parastiedica n. sp. were subspheroid, 27.4 X 25.5 (25-30 X 22-28) micron; micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts, pointed at both ends, were 18.3 X 10.4 (16-20 X 9-11) micron; Stieda, sub-, and parastieda bodies were present as was a sporocyst residuum. This species was found in 2 of 18 (11%) hosts.(ABSTRACT TRUNCATED AT 250 WORDS)     

Eimeria reedi sp. n. and E. chobotari sp. n. (Protozoa: Eimeriidae) from Heteromyid Rodents

SYNOPSIS The oocysts of 2 new species of Eimeria are described from heteromyid rodents. The subspherical to ellipsoidal cocysts of Eimeria reedi from Perognathus formosus averaged 22.6 by 20.8 μ. An ellipsoidal polar granule was present. The oocyst residuum was a single globule about 8 μ in diameter. The ovoid sporocysts averaged 10.9 by 7.7 μ and had a residuum of 6-12 granules. A Stieda body and substiedal body were present in the sporocysts. Eimeria chobotari oocysts, from Dipodomys merriami and D. microps , were ellipsoidal to ovoid and averaged 47.7 by 35.2 μ. No polar granule was seen. The large oocyst residuum consisted of several granules of various sizes. The ovoid sporocysts averaged 15.8 by 11.3 μ and had a residuum of many small granules. A Stieda body and substiedal body were present in the sporocysts.