鸡疟原虫成孢子细胞和子孢子形成的电镜观察

应用扫描和透射电镜观察鸡疟原虫在白纹伊蚊体内发育的成孢子细胞和子孢子形成过程的形态特征。扫描电镜观察,成孢子细胞形状多样,呈长形、圆形、椭圆形,常为不规则形。子孢子较其他疟原虫短粗、稍弯曲。透射电镜观察可见在成孢子细胞形成初期,首先在卵囊被膜下层出现许多液泡,这些液泡逐渐增大且伸入卵囊的胞质内,而形成许多裂缝;继之,裂缝互相连接,而将卵囊胞质割裂为许多团块状成孢子细胞。子孢子芽从成孢子细胞体表面长出,逐渐发育成长形子孢子,此时可见其前端内部出现棒状体和微线体等细胞器。     

共聚焦激光扫描显微镜对约氏疟原虫卵囊、子孢子胞内Ca~(2 )的检测方法研究

本文报告建立用共聚焦激光扫描显微镜 (CLSM)观察约氏疟原虫卵囊、子孢子胞内游离Ca2 的分布和实验检测方法。以荧光染料Fluo 3作胞内Ca2 指示剂 ,结果显示 3μ、 4μmol/LFluo 3/Am同时加入 1μl/ml 2 5 %PluronicF 12 7在 37℃恒温下 ,分别孵育分离的约氏疟原虫子孢子、卵囊 6 0min即可达到最佳的负载效果。Fluo 3/Am浓度的提高和孵育时间延长只能增加背景染色 ,若降低孵育浓度和缩短孵育时间则难于达到最佳的负载效果。子孢子胞内游离Ca2 分布均匀 ;而卵囊内游离Ca2 分布不均匀 ,成块状分布 ,囊壁无荧光。研究表明应用Fluo 3/AM和PluronicF 12 7结合CLSM技术是研究疟原虫卵囊、子孢子胞浆内游离Ca2 的准确、灵敏的方法之一     

共聚焦激光扫描显微镜对约氏疟原虫卵囊、子孢子胞内Ca^2＋的检测方法研究

本文报告建立用共聚焦激光扫描显微镜（CLSM）观察约氏疟原虫卵囊,子孢子胞内游离Ca^2 的分布和实验检测方法,以荧光染料下Fluo-3作胞内Ca^2 指示剂,结果显示3u,4umol/L Fluo-3/Am同时加入1ul/ml25%PluronicF-127在37℃恒温下,分别孵育分离的约氏疟原虫子孢子,卵囊60min即可达到最佳的负载效果,子孢子胞内游离C^2 分布均匀,而卵囊内游离C^2 a分布不均匀,成块状分布,囊壁无效荧光,研究表明应用Fluo-3/AM和PluronicF-127结合CLSM技术是研究疟原虫卵囊,子孢子胞浆内游离Ca^2 的准确,灵敏的方法之一。     

食蟹猴疟原虫晚期卵囊、成孢子细胞与子孢子扫描电镜观察

本研究对实验感染食蟹猴疟原虫后9、10和14天的斯氏按蚊阳性蚊胃,进行扫描电镜观察。晚期(分化的)卵囊表面呈现凹凸不平皱褶。成孢子细胞体常呈圆球形或椭圆形,表面光滑,子孢子芽从其表面长出。子孢子体细长,稍弯曲,体表光滑;虫体前部较细,顶端稍平,后部稍粗,末端钝圆。本文对成孢子细胞不同发育阶段的形态予以较详细描述,对子孢子出囊方式做了初步讨论。     

萝卜CMS不育系与保持系小孢子发生的细胞学研究

研究了萝卜胞质雄性不育系A2、A4及其相应保持系B2、B4的小孢子发生与花药壁发育的细胞学特征.结果表明,不育系A2的绒毡层细胞在四分体时期出现异常,小液泡增多,至单核期汇合形成大液泡,绒毡层细胞异常膨大;小孢子外壁染色浅,细胞壁受到破坏,最后与绒毡层一同降解.不育系A4在减数分裂期即表现出异常,绒毡层异常肥大;花药发育后期,小孢子外壁亦染色较浅;绒毡层细胞融合形成细胞团块侵入药室挤压小孢子,两者一同降解.     

同源四倍体水稻小孢子发生的超微结构

利用透射电子显微镜对同源四倍体水稻小孢子母细胞减数分裂前及期间的超微结构进行观察,结果发现:(1)减数分裂前及减数分裂早期,小孢子母细胞核糖体密度高,线粒体、质体等细胞器丰富;粗线期核糖体密度显著下降,线粒体、质体等细胞器数量减少;终变期核糖体密度逐渐恢复到减数分裂前状态,而其他细胞器的数量除在二分体时期出现短暂的回升,终变期以后的时期均较少.(2)小孢子母细胞间的连接在小孢子母细胞时期以典型的胞间连丝为主,进入细线期,胞间连丝数量显著减少,宽孔道的胞质通道逐渐增多,粗线期小孢子母细胞间基本通过胞质通道连接,终变期小孢子母细胞间完全分离.(3)随着减数分裂的进行,药壁四层细胞逐渐液泡化,绒毡层细胞中部分小液泡融合成大液泡,形成胞内"空腔";药室内壁细胞出现大量的具有叶绿体结构特征的质体,内含丰富的淀粉粒,到了四分体时期质体数量及内含的淀粉粒显著减少.     

鸡疟原虫孢子体超微结构的观察

埃及伊蚊感染鸡疟原虫（Plasmodium gallinaceum）18天后解剖,电镜下观察唾液腺内孢子体的形态。孢子体长7μm、宽0.8μm;复合膜由一层外膜、二层内膜及膜下微管组成。发达的膜下微管与孢子体重要的运动功能有关。细胞核约位于正中。胞质较均一,有时有空泡存在,胞质中有散在核糖体,未观察到内质网。孢子体有胞口。发达的棒状体及众多的微线体,可能与孢子体需侵入媒介唾液腺细胞、尔后再侵入鸟类宿主中胚层细胞有关。因而,任何作用于棒状体、微线体并导致其结构及功能变化的药物,都将影响甚至阻断孢子体对宿主细胞的入侵,这就为疟疾的药物预防提供重要理论依据。     

黄胫小车蝗卵子发生及卵母细胞凋亡的显微观察

对黄胫小车蝗(Oedaleus infernalis)卵子发生过程和卵母细胞凋亡进行显微观察。结果表明,黄胫小车蝗卵子发生可明显分为3个时期10个阶段,即卵黄发生前期、卵黄发生期和卵壳形成期。第1阶段,卵母细胞位于卵原区,经历减数第一次分裂;第2阶段,卵母细胞核内染色体解体成网状,滤泡细胞稀疏地排列在卵母细胞周围;第3阶段,滤泡细胞扁平状,在卵母细胞周围排成一层;第4阶段,滤泡细胞呈立方形排在卵母细胞周围;第5阶段,滤泡细胞呈长柱形排在卵母细胞周围,滤泡细胞之间、滤泡细胞与卵母细胞之间出现空隙;第6阶段,卵母细胞边缘开始出现卵黄颗粒;第7阶段,卵母细胞中沉积大量卵黄,胚泡破裂;第8阶段,滤泡细胞分泌卵黄膜包围卵黄物质;第9阶段,滤泡细胞分泌卵壳;第10阶段,卵壳分泌结束,卵子发育成熟。卵母细胞发育过程中的凋亡发生在卵黄发生前期,主要表现为滤泡细胞向卵母细胞内折叠,胞质呈团块状等特征。     

蜡梅小孢子发生和花粉形成的研究

通过对蜡梅小孢子发生和花粉形成的研究,结果表明：蜡梅幼小花药中的多列孢原细胞经造孢细胞发良为小孢子母细胞。减数分裂为同时型。四分体呈四面体型排列,同时观察了小孢子在发育过程中液泡的动态变化。成熟花粉为２－细胞型。花药壁的发育为双子叶型。花药壁由５－６层细胞组成,腺质绒毡层。花粉具有异型性现象。     

毛钩藤的小孢子发生和雄配子体发育

在光学显微镜和透射电镜下观察了毛钩藤(Uncaria hirsuta Havil.)的小孢子发生和雄配子体发育过程.结果表明,毛钩藤花两性,具5枚雄蕊,花药4室,花药壁由表皮、药室内壁、中层和绒毡层组成,花药开裂时,药室内壁高度纤维化带状加厚.花药壁的发育方式属于双子叶型,小孢子母细胞减数分裂的胞质分裂为同时型.小孢子在四分体时期开始沉积花粉外壁,小孢子大液泡化时期开始沉积花粉内壁.成熟花粉为2-细胞型.毛钩藤的花粉发育特征和茜草科植物基本一致.毛钩藤绒毡层属于分泌型,双重起源,分别起源于次生周缘层和药隔细胞.小孢子发育早期绒毡层开始降解并分泌形成大量乌氏体,花药开裂时绒毡层完全消失,剩下少量乌氏体.小孢子早期内壁加厚突出形成,小孢子细胞核分裂以后内壁加厚开始脱落,花药开裂时,只剩下少量的内壁加厚突出.初步推测,内壁加厚突出与乌氏体共同作用为雄配子体的发育提供营养物质.     

Transformation of the Plasmodium cynomolgi Oocyst1

SYNOPSIS. The transformation of the P. cynomolgi oocyst into definitive sporozoite forms occurs 8–10 days after an infective blood meal by Anopheles, stephensi mosquitoes. Vacuolization divides the oocyst cytoplasm into sporoblast sub-units from which sporozoites bud. The role of sporoblast nuclear and cytoplasmic components in the complex differentiative process is discussed.     

THE TRANSFORMATION OF THE PLASMODIUM GALLINACEUM OOCYST IN AEDES AEGYPTI MOSQUITOES

Sporoblast and sporozoite formation from oocysts of the avian malarial parasite, Plasmodium gallinaceum, after the seventh day of infection in Aedes aegypti mosquitoes offers an interesting example of differentiation involving the appearance and modification of several cellular components. Sporoblast formation is preceded by (a) invaginations of the oocyst capsule into the oocyst cytoplasm, (b) subcapsular vacuolization and cleft formation, (c) the appearance of small tufts of capsule material on the previously noted invaginations, and (d) linear dense areas located just below the oocyst plasma membrane which predetermine the site of emerging sporozoites from the sporoblast. The subcapsular clefts subdivide the once-solid oocyst into sporoblast peninsulae. Within the sporoblast, nuclei migrate from the random distribution seen in the solid oocyst and come to lie at the periphery of the sporoblast just below the linear dense areas noted in the earlier stage. A typical nuclear fiber apparatus occurs in most of the nuclei seen in random sections at this stage although such a fiber apparatus may occasionally be seen in the solid oocyst stage. The nucleus, its associated fiber apparatus, and the overlying dense area appear to induce the onset of sporozoite budding from the sporoblast as well as the formation of the sporozoite pellicular complex and the paired organelle precursor. Several mitochondria are present in each sporozoite, in contrast to the single mitochondrion seen in the merozoites of the erythrocytic and exoerythrocytic stages of avian malaria infection. The paired organelles and associated dense inclusion bodies are formed by condensation of an irregular meshwork of membrane-bound, coarse, dense material. The nature of small, particulate cytoplasmic inclusions is described.     

Susceptibility of Anopheles campestris-like and Anopheles barbirostris species complexes to Plasmodium falciparum and Plasmodium vivax in Thailand

Nine colonies of five sibling species members of Anopheles barbirostris complexes were experimentally infected with Plasmodium falciparum and Plasmodium vivax. They were then dissected eight and 14 days after feeding for oocyst and sporozoite rates, respectively, and compared with Anopheles cracens. The results revealed that Anopheles campestris-like Forms E (Chiang Mai) and F (Udon Thani) as well as An. barbirostris species A3 and A4 were non-potential vectors for P. falciparum because 0% oocyst rates were obtained, in comparison to the 86.67-100% oocyst rates recovered from An. cracens. Likewise, An. campestris-like Forms E (Sa Kaeo) and F (Ayuttaya), as well as An. barbirostris species A4, were non-potential vectors for P. vivax because 0% sporozoite rates were obtained, in comparison to the 85.71-92.31% sporozoite rates recovered from An. cracens. An. barbirostris species A1, A2 and A3 were low potential vectors for P. vivax because 9.09%, 6.67% and 11.76% sporozoite rates were obtained, respectively, in comparison to the 85.71-92.31% sporozoite rates recovered from An. cracens. An. campestris-like Forms B and E (Chiang Mai) were high-potential vectors for P. vivax because 66.67% and 64.29% sporozoite rates were obtained, respectively, in comparison to 90% sporozoite rates recovered from An. cracens.     

Plasmodium vivax:A Monoclonal Antibody Recognizes a Circumsporozoite Protein Precursor on the Sporozoite Surface

Gonzalez-Ceron, L., Rodriguez, M. H., Wirtz, R. A., Sina, B. J., Palomeque, O. L., Nettel, J. A., and Tsutsumi, V. 1998.Plasmodium vivax:A monoclonal antibody recognizes a circumsporozoite protein precursor on the sporozoite surface.Experimental Parasitology90, 203–211. The major surface circumsporozoite (CS) proteins are known to play a role in malaria sporozoite development and invasion of invertebrate and vertebrate host cells.Plasmodium vivaxCS protein processing during mosquito midgut oocyst and salivary gland sporozoite development was studied using monoclonal antibodies which recognize different CS protein epitopes. Monoclonal antibodies which react with the CS amino acid repeat sequences by ELISA recognized a 50-kDa precursor protein in immature oocyst and additional 47- and 42-kDa proteins in older oocysts. A 42-kDa CS protein was detected after initial sporozoite invasion of mosquito salivary glands and an additional 50-kDa precursor CS protein observed later in infected salivary glands. These data confirm previous results with otherPlasmodiumspecies, in which more CS protein precursors were detected in oocysts than in salivary gland sporozoites. A monoclonal antibody (PvPCS) was characterized which reacts with an epitope found only in the 50-kDa precursor CS protein. PvPCS reacted with allP. vivaxsporozoite strains tested by indirect immunofluorescent assay, homogeneously staining the sporozoite periphery with much lower intensity than that produced by anti-CS repeat antibodies. Immunoelectron microscopy using PvPCS showed that the CS protein precursor was associated with peripheral cytoplasmic vacuoles and membranes of sporoblast and budding sporozoites in development oocysts. In salivary gland sporozoites, the CS protein precursor was primarily associated with micronemes and sporozoite membranes. Our results suggest that the 50-kDa CS protein precursor is synthesized intracellularly and secreted on the membrane surface, where it is proteolytically processed to form the 42-kDa mature CS protein. These data indicate that differences in CS protein processing in oocyst and salivary gland sporozoites development may occur.     

Scanning electron microscopic observations of the oocyst, sporoblast and sporozoite of Plasmodium yoelii yoelii

Scanning electron microscopy was used to study the surface characteristics of the oocyst, sporoblast and sporozoite of Plasmodium yoelii yoelii. Observations were made of the sporogonic stages of 6-12 day infections of the malaria parasite in Anopheles stephensi. Oocyst and sporoblast development were not synchronous. The surface of the undifferentiated (early stage) oocyst appeared smooth, whereas that of differentiated (late stage) oocysts were rough or wrinkled. The wall of the differentiated oocysts showed numerous micropores at higher magnification (x15,000-20,000) the biological significance of which is not known. Small, bud-like satellite bodies were seen attached to some oocysts. Various forms of different stages of the sporoblast were described. Sporozoite budding took place on the surface of the sporoblast body. The sporozoite was elongate, curved and with a blunt anterior end.     

Some Observations on the Sporogonic Cycle of Plasmodium schwetzi, P. vivax and P. ovale in Five Species of Anopheles

SYNOPSIS. The sporogonic cycles of Plasmodium schwetzi, P. vivax and P. ovale are distinctly different with regard to rate of sporozoite development and median oocyst diameter. Differences in the oocyst structure, particularly with regard to the presence of inclusions in those of P. schwetzi , were noted. These findings tend to support the continued designation of these parasites as distinct and separate species.     

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.     

Studies on the Santa Lucia (El Salvador) strain of Plasmodium falciparum in Aotus trivirgatus monkeys.

The Santa Lucia strain of Plasmodium falciparum was isolated from El Salvador, Central America, and established in Aotus trivirgatus monkeys. Transmission from monkey to monkey via the bites of infected Anopheles freeborni, A. maculatus, and A, albimanus mosquitoes was obtained in 20 of 27 attempts. Prepatent periods in the monkeys ranged from 17 to 46 days with a mean of 24.3 days. Parasitemias and mortality were higher following sporozoite inoculation into animals which had been previously infected with P. vivax than in those with no previous malaria experience. Monkeys previously infected with P. vivax and P. cynomolgi had lower maximum parasitemias than those previously infected with P. vivax only.     

Electron Microscopic and Histochemical Studies of Sporozoite Formation in Plasmodium berghei*

SYNOPSIS. Observations were made on the differentiation of fine structure during sporogonic development of Plasmodium berghei. The oocyst in the process of sporozoite formation is an encapsulated structure 30-40 μ in diameter. It typically develops while in an extracellular position, attached to the basement membrane of the mosquito midgut and projecting into the mosquito hemocoel. Occasionally, however, ookinetes passing thru the midgut epithelial cells may become impacted within a cell so that the resulting oocyst develops intracellularly. Each oocyst has a large differentiating region, the sporoblastoid body. This body contains large dividing nuclei which are Feulgenpositive, and a cytoplasm which includes mitochondria, dense rodlike structures, cytoplasmic membranes, cisternae and vacuolar structures, Golgi material, and ribosomes which are both free and membrane-associated. Sporozoite budding takes place along the surface of the sporoblastoid body. Bits of a new membrane condense under the plasma membrane which bounds the sporoblastoid body. These 2-membraned sites then bulge out, continue to elongate, and eventually become sporozoites. The various nuclear and cytoplasmic components of the sporoblastoid body are passed into the sporozoites during their elongation. In addition, the sporozoite develops a system of elogate, subpellicular microtubules, possibly contractile in function. The pellicle of the sporozoite is broken by an opening, the cytostome (micropyle). The anterior end is truncate.     

Six new Eimeria species from vespertilionid bats of North America.

Twenty species of bats (Molossidae, Vespertilionidae) were collected from California, New Mexico, Oregon, South Carolina, Utah, and Baja California Norte (Mexico), and 29 of 404 (7%) animals, including Antrozous pallidus, Eptesicus fuscus, Myotis auriculus, Myotis californicus, Myotis ciliolabrum, Myotis evotis, Myotis lucifugus, Myotis thysanodes, Myotis vivesi, Myotis volans, Myotis yumanensis, and Nycticeius humeralis were infected with Eimeria spp., which represent 6 new species. Sporulated oocysts of a new species from A. pallidus are subspheroidal, 24.8 x 21.6 (22-27 x 19-24) microm with a polar granule and a large globular residuum. The oocyst wall is sculptured, with 2 layers, approximately 1.5 thick. Ovoidal sporocysts are 11.5 x 7.8 (9-13 x 7-10) microm, with Stieda body and residuum of many large granules. Sporulated oocysts of a new species from M. californicus are subspheroidal, 20.7 x 18.2 (19-23 x 16-20) microm, with 1-7 tiny polar granules, but without oocyst residuum. The oocyst wall is rough, with 2 layers, approximately 1.4 thick. Ovoidal sporocysts are 11.2 x 7.3 (10-12 x 7-8) microm, with Stieda body and a globular residuum. Sporulated oocysts of a second new species from M. californicus are subspheroidal, 23.1 x 20.7 (20-26 x 19-23) microm, with residuum and 1 polar granule, but a micropyle is absent. The oocyst wall is rough with 2 layers, approximately 1.5 thick. Ovoidal sporocysts are 12.5 x 7.2 (11-14 x 7-8) microm, with a Stieda body and residuum. Sporulated oocysts of a new species from M. ciliolabrum are subspheroidal, 24.9 x 20.1 (18-27 x 17-23) microm, with 1-2 polar granules, but without micropyle and residuum. The oocyst wall is rough with 2 layers, approximately 1.5 thick. Ellipsoidal sporocysts are 12.5 x 9.0 (8-14 x 7-10) microm, with Stieda and substieda bodies and residuum. Sporulated oocysts of a new species from M. evotis are subspheroidal, 21.3 x 18.6 (20-24 x 15-20) microm, with a prominent polar granule, but without micropyle and residuum. The oocyst wall is smooth with 2 layers, approximately 1.0 thick. Ovoidal sporocysts are 12.2 x 8.0 (11-13 x 7.5-9) microm, with Stieda and substieda bodies and residuum. Sporulated oocysts of the new species from N. humeralis are subspheroidal, 22.4 x 18 (21-24 x 17-20) microm, with 1-3 polar granules, but residuum and micropyle are absent. The oocyst wall is lightly sculptured with 2 layers, approximately 1.4 thick. Ovoidal sporocysts are 10.9 x 7.7 (9-12 x 6-8) microm, with Stieda body and residuum. Sporulated oocysts of E. pilarensis Scott and Duszynski, 1997 and those of at least 12 other morphological forms were seen in the other infected bats; these latter forms were seen in too few numbers to be adequately described as new species.