广东艾美虫发育的研究

广东艾美虫寄生于乌鳢的消化道,其发育周期可分为裂体生殖、配子生殖和孢子生殖三个阶段。这三个阶段均在一个寄主体内完成。裂体生殖和配子生殖发生在幽门盲囊和前肠上皮细胞核之上方。成熟裂殖体为球形或椭圆形,含有8—14个香蕉形裂殖子。大配子的整个发育过程没有发现嗜伊红颗粒,嗜碱性颗粒在卵囊壁形成前后发生变化。成熟小配子母细胞内有许多新月形的小配子。孢子生殖在幽门盲囊和整条肠管内进行。       

中华血簇虫的生活史研究(复顶门:孢子纲) Ⅱ.中华血簇虫在中华鳖中的发育

中华血簇虫在其无脊椎动物寄主中的发育已另有文描述。这里报道的是中华血簇虫在中华鳖中的发育。这一时期包括三个阶段:组织细胞内裂体增殖、深部血红细胞内的裂体增殖和外周血红细胞内的裂体增殖。组织细胞内裂殖体产生14—32个裂殖子。深部血红细胞内的裂殖体分为两类:一类是X裂殖体,它产生14—18个小裂殖子;另一类是Y裂殖体,它产生4—6个大裂殖子。外周血红细胞内的初期裂殖体可产生多至14个裂殖子,而随后的裂体增殖却产生越来越少的裂殖子,且裂殖体和裂殖子的大小也渐趋变小。外周血晚期的裂殖体只形成2个裂殖子。配子母细胞来源于Y裂殖子。营养体是由上一代裂殖子向下一代裂殖体发育的中间时期。     

柔嫩艾美耳球虫配子发生的超微结构

利用透射电镜对柔嫩艾美耳球虫配子生殖阶段的超微结构进行了,大配子体和小配子体于相邻的宿主肠上皮细胞内相产生,由末代裂殖子入后,长大变圆而形成,小配子的形成为直接分化型,首先细胞核分裂成为多核体,随后细胞核向周边移动,然后紧靠细胞处的限制向外突出,临近突出部位的限制膜下陷,在核上方形成中心粒,中心粒发育为基粒,鞭毛中的微管和附着微管,早期形成的小配子仍与小配子体的殖体相连,成熟的小配子与配子体分离,外型香蕉状,外被单位膜,内有一电子结构十分致密的细胞核,核的头端侧面有一个巨大的线粒体,小配子有鞭毛2根,每根鞭毛内有微管,组成为9＋2结构,此外,小配子至少有6根附着微管,大配子体和大配子外被单位膜,内部形成大量的成囊体1和成囊体2,并有大量的支链淀粉和脂肪体,中央有一个细胞核,卵囊臂有5层,细胞核位于细胞中央,细胞内有大量的支链淀粉和脂肪体。     

水牛枯氏住肉孢子虫在犬体内发育史的研究

10只一月龄幼犬用于研究水牛枯氏住肉孢子虫的内生发育史,其中9只接种包囊,1只不接种作对照。感染后12～24小时,缓殖子进入小肠绒毛内形成大配子(4.73×3.82μm)和小配子(5.20×3.92μm),2—5天发育成卵囊;8—12天卵囊孢子化;18天卵囊孢子化完成。成熟卵囊内含2个孢予囊。孢子囊15.24×9.95μm,内有4个子孢子和成团或散状的残余体。整个发育过程在小肠绒毛(尤其是回肠上段)的基底膜与固有层的带虫空泡内进行。     

2n雄配子的无融合生殖披碱草（Elymus rectisetus）减数分裂行为研究

用能自然产生2n雄配子的无融合生殖六倍体披碱草为材料,研究了无融合生殖披碱草的减数分裂行为。结果表明：2n雄配子的无融合生殖披碱草减数分裂行为较为异常。在间期细胞内经常有微核存在;在前期I,有倒位环、多介染色体等异常现象;在中期I,染色体有不等分裂的倾向;在后期I和末期I,有染色体的单极分裂（63.9%）、染色走向一极的倾向（21.7%）和多极分裂（3.3%）等现象,单极分裂的结果是不减数配子的产生。在减数分裂Ⅱ中,有高比例的二分子孢子（55.7%）和三分子孢子（23.7%）的形成,二分子孢子在发育后期也直接导致了2n配子的产生。因此,染色体的单极分裂、二分体和三分体形成这3种方式是披碱草2n雄配子的形成的主要途径。     

贝氏隐孢子虫在北京鸭体内发育的超微结构研究

贝氏隐孢子虫各期虫体均位于宿主粘膜上皮细胞的带虫空泡中。在虫体与上皮细胞接触处,虫体表膜反复折迭形成营养器。子孢子或裂殖子与粘膜上皮细胞接触后,逐步过渡为球形的滋养体;滋养体经2—3次核分裂、产生含4或8个裂殖子的两代裂殖体,裂殖体以外出芽方式产生裂殖子;裂殖子无微孔,顶端表皮形成3—4个环嵴,裂殖子进一步发育成为配子体;大配子体含有两种类型的成囊体。小配子呈楔形,无鞭毛和顶体,有一个致密的长椭圆形细胞核,小配子表膜内侧有9根膜下微管;孢子化卵囊内含四个裸露的子孢子和一个大残体。本文是有关鸭体内隐孢子虫超微结构的首次报导。     

金粉蕨配子体发育及其无配子生殖的研究

采用光学显微镜和半薄切片技术对金粉蕨(Onychium siliculosum(Desv.)C.Chr.)的配子体发育及其无配子生殖进行了研究。金粉蕨孢子黄褐色,具三裂缝,孢子萌发时先产生假根,然后长出原叶体细胞,萌发类型为书带蕨型。原叶体细胞经丝状体、片状体发育为原叶体,发育类型为水蕨型。金粉蕨配子体分枝发达,成熟配子体呈丛状。多次培养表明金粉蕨只能产生精子器,不能产生颈卵器,为专性无配子生殖。半薄切片观察表明配子体生长点下方的细胞经分裂产生一群小型细胞,由它们发育为无配子生殖胚。小型细胞先分化出无性胚芽,再分化出胚根。胚芽与胚根,胚与配子体之间由分化出的导管连接。     

四倍体双穗雀稗兼性无孢子生殖的研究

研究了四倍体双穗雀稗(Paspalum distichum L)无孢子生殖胚囊、胚胎发育以及假受精特点。当其大孢子母细胞发育至四分体阶段时,大多数情况下会发生四分体退化,同时有多个特化珠心细胞发育为1—3个无孢子生殖胚囊的现象。成熟无孢子生殖胚囊一般3核,包括1个卵细胞和2个极核。卵细胞在抽穗前就能自发分裂形成原胚团,而极核则在抽穗和传粉后参与假受精形成胚乳。当胚珠内存在多个无孢子生殖胚囊时,只是靠近珠孔端的1个无孢子生殖胚囊内的极核与精核结合,而其它的并不参与。种子成熟后出现很低频率的二胚苗。此外,还能观察到少量的有性生殖胚囊的发育以及有性生殖胚囊和无孢子生殖胚囊在同一胚珠中的发育现象,因此判断该类群为兼性无孢子生殖体。     

肠艾美耳球虫（Eimeria intestinalis）内生发育阶段超微结构研究：Ⅱ…

肠艾美耳球虫大配子体带虫空泡内有许多泡内小管。大配子体外被单层限制膜,核大,有一明显的核仁。两种成囊颗粒几乎同时出现,发育中的大配子体胞质中出现多糖和脂肪小体,并逐渐增多,大配子表面能两层膜。卵囊壁形成的标志是大配子表面的两层膜松弛地向带虫空泡内推移,成囊颗粒Ｉ形成卵囊壁的外层,成囊粒Ⅱ的物质形成卵囊壁的内层,在内层之下又有一颗粒层的形成和消失。     

肠艾美耳球虫(Eimeria intestinalis)内生发育阶段超微结构研究:Ⅱ.大配子的发育及卵囊壁的形成

肠艾美耳球虫(Eimeria intestinalis)大配子体(Macrogametocyte)带虫空泡内有许多泡内小管。大配子体外被单层限制膜,核大,有一明显的核仁。两种成囊颗粒几乎同时出现。发育中的大配子体胞质中出现多糖和脂肪小体,并逐渐增多。大配子表面被有两层膜。卵囊壁形成的标志是大配子表面的两层膜松弛地向带虫空泡内推移,成囊颗粒Ⅰ形成卵囊壁的外层,成囊颗粒Ⅰ的物质形成卵囊壁的内层,在内层之下又有一颗粒层的形成和消失。     

Observations on Haemogregarina balli sp. n. from the Common Snapping Turtle,Chelydra serpentina*

SYNOPSIS. Haemogregarina balli sp. n. is described from the blood and organs of the common snapping turtle Chelydra serpentina serpentina and from the gastric and intestinal ceca of the presumed invertebrate hosts, the leeches Placobdella parasitica and Placobdella ornata. In the peripheral blood of the turtle, male and female gametocytes and immature erythrocytic schizonts are found within erythrocytes. The maturation of erythrocytic schizonts containing 6–8 merozoites is recorded from liver imprints. Schizonts with 13–25 merozoites are found in various cells of the liver, lung and spleen. In the gastric ceca of the leeches the host erythrocytes are digested, releasing the gametocytes and immature erythrocytic schizonts. Immature erythrocytic schizonts degenerate. Association of the gametocytes occurs in the intestinal ceca. The microgametocyte apparently gives rise to 4 nonmotile microgametes, one of which fertilizes the macrogamete while the other remain as condensed, residual nuclei on the periphery of the developing oocyst. The oocyst increases in size with maturity. A mature oocyst produces 8 sporozoites from a single germinal center. Sporozoites liberated from the oocyst are found in the tissues of the leech. Transovarial transmission of the parasite does not occur in the turtle. Attempts at experimental transmission failed. Previously unfed (control) leeches were negative for the parasite. Haemogregarina balli is compared with other haemogregarines described from C. serpentina. Features of species of Haemogregarina and Hepatozoon as well as the taxonomy of these genera are discussed.     

A new hemogregarine from marine fishes.

Haemogregarina uncinata sp. n. is described from the blood of 2 marine eelpouts, Lycodes lavalaei and Lycodes vahlii (Perciformes: Zoarcidae). Erythrocytic schizogony occurred in peripheral and cardiac blood, but mature schizonts were restricted to the latter site. Mature and rupturing schizonts contained 10 to 30 merozoites, which were short and thick in small schizonts while slender and long in larger schizonts. Gametocytes developed in mature erythrocytes and displayed morphologic and morphometric characters that distinguished them from other species described. Syzygy and gamete formation occurred in the gut of a leech, Johanssonia sp. Each microgametocyte produced up to 4 apparently nonflagellated gametes. Oocysts developed intracellularly in the epithelial wall of the intestine and at maturity produced under 100 sporozoites from (apparently) several germinal centers. Sporozoites subsequently migrated to the probosces of the leeches. The failure to transmit the parasite to a sculpin (Myoxocephalus octodecemspinosus) and 3 Atlantic cod (Gadus morhua) via regurgitation by the leeches might be indicative of host specificity.     

Development of Babesiosoma stableri (Dactylosomatidae; Adeleida; Apicomplexa) in its Leech Vector (Batracobdella picta) and the Relationship of the Dactylosomatids to the Piroplasms of Higher Vertebrates

The sporogonic and merogonic development of Babesiosoma stableri Schmittner & McGhee, 1961 within its definitive host and vector, a leech Batracobdella picta (Verrill, 1872), was studied by light and electron microscopy. Gamonts released from frog erythrocytes in the blood meal of the leech associated in syzygy and fused; the gamonts were isogamous and only 1 microgamete was formed. The ultrastructural appearance of the resulting zygote was similar to that of the gamonts, but it was larger. The zygote had an apical complex (including a polar ring, conoid and 2 pre-conoidal rings and micronemes, but no recognizable rhoptries), triple-membraned pellicle, about 40 subpellicular microtubules and prominent stores of amylopectin. Zygotes penetrated the cells of the intestine and underwent sporogony directly within the cytosplasm of the ieech epithelial cell without the formation of a parasitophorous vacuole. Eight sporozoites budded simultaneously around the periphery of an irregularly shaped oocyst. No oocyst wall was formed. Each sporozoite had a complete apical complex (including rhoptries), abundant amylopectin inclusions and a triple-membraned pellicle with about 32 subpellicular microtubules. The sporozoites initiated merogonic replication primarily within the salivary cells of the leech although other tissues, such as muscle, were infected. Each meront produced 4 merozoites by simultaneous budding, forming a cruciform meront typical of the intraerythrocytic development of this parasite. The meront was located directly within the cytoplasm of the host cell. Merozoites, with abundant amylopectin, had a complete apical complex and triple-membraned pellicle with about 40 subpellicular microtubules. The merozoites either initiated a further cycle of replication, or they moved into the ductules of the leech salivary cells which extend to the tip of the proboscis. Observations on gametogenesis. syngamy and sporogony of B. stableri in its leech host indicate that the family Dactylosomatidae should be placed in the suborder Adeleina (Eucoccidiida: Apicomplexa). Babesiosoma stableri was transmitted to uninfected frogs (Rana spp.) by the bite of infected leeches. Prepatent periods ranged from 26 to 38 days at 25° C. Despite a directed search in laboratory reared tadpoles which had each been injected intraperitoneally with 150,000 merozoites, no pre-erythrocytic developmental stages were observed. Similarities in their biology suggest close phylogenetic affinities of the dactylosomatids, and other adeleid blood parasites, with the piroplasms of higher vertebrates.     

Distinct malaria parasite sporozoites reveal transcriptional changes that cause differential tissue infection competence in the mosquito vector and mammalian host

The malaria parasite sporozoite transmission stage develops and differentiates within parasite oocysts on the Anopheles mosquito midgut. Successful inoculation of the parasite into a mammalian host is critically dependent on the sporozoite's ability to first infect the mosquito salivary glands. Remarkable changes in tissue infection competence are observed as the sporozoites transit from the midgut oocysts to the salivary glands. Our microarray analysis shows that compared to oocyst sporozoites, salivary gland sporozoites upregulate expression of at least 124 unique genes. Conversely, oocyst sporozoites show upregulation of at least 47 genes (upregulated in oocyst sporozoites [UOS genes]) before they infect the salivary glands. Targeted gene deletion of UOS3, encoding a putative transmembrane protein with a thrombospondin repeat that localizes to the sporozoite secretory organelles, rendered oocyst sporozoites unable to infect the mosquito salivary glands but maintained the parasites' liver infection competence. This phenotype demonstrates the significance of differential UOS expression. Thus, the UIS-UOS gene classification provides a framework to elucidate the infectivity and transmission success of Plasmodium sporozoites on a whole-genome scale. Genes identified herein might represent targets for vector-based transmission blocking strategies (UOS genes), as well as strategies that prevent mammalian host infection (UIS genes).     

Levels of circumsporozoite protein in the Plasmodium oocyst determine sporozoite morphology

The sporozoite stage of the Plasmodium parasite is formed by budding from a multinucleate oocyst in the mosquito midgut. During their life, sporozoites must infect the salivary glands of the mosquito vector and the liver of the mammalian host; both events depend on the major sporozoite surface protein, the circumsporozoite protein (CS). We previously reported that Plasmodium berghei oocysts in which the CS gene is inactivated do not form sporozoites. Here, we analyzed the ultrastructure of P.berghei oocyst differentiation in the wild type, recombinants that do not produce or produce reduced amounts of CS, and corresponding complemented clones. The results indicate that CS is essential for establishing polarity in the oocyst. The amounts of CS protein correlate with the extent of development of the inner membranes and associated microtubules underneath the oocyst outer membrane, which normally demarcate focal budding sites. This is a first example of a protein controlling both morphogenesis and infectivity of a parasite stage.     

Experimental observations on the specificity of Apatemon (Australapatemon) minor (Yamaguti 1933) (Digenea: Strigeidae) toward leech (Hirudinea) second intermediate hosts

The infectivity of Apatemon (Australapatemon) minor cercariae to 7 species of freshwater leech was examined under laboratory conditions. The leech species exposed to infection were; Erpobdella octoculata, Helobdella stagnalis, Glossiphonia heteroclita, Glossiphonia complanata, Hemiclepsis marginata, Piscicola geometra, and Theromyzon tessulatum. Five species were capable of acting as second intermediate hosts. Despite this broad specificity only Erpobdella octoculata and Helobdella stagnalis could be considered to show a high degree of compatibility with the parasite. In these 2 leech species more than 38% of the cercariae successfully penetrated and established metacercarial cyst infections. The leeches Piscicola geometra and Theromyzon tessulatum did not become infected. Close parallels were found between the species order of host utilization determined experimentally and that revealed by a range of field surveys in Britain, Eastern Europe and Russia. This suggests that the order of second intermediate host utilization in the natural environment has, at least in part, a physiological basis. The degree to which different leeches are utilized as hosts may relate to the opportunity which they afford for transmission of A. minor to wildfowl definitive hosts.     

Development of Eimeria tenella in MDBK cell culture with a note on enhancing effect of preincubation with chicken spleen cells

Eimeria tenella, an intracellular protozoan parasite infecting the epithelial cells of the ceca of chickens, causes severe diarrhea and bleeding that can lead its host to death. It is of interest that E. tenella first penetrate into the mucosal intraepithelial lymphocytes (IEL) before they parasitize crypt or villous epithelial cells. This in vitro study was undertaken to know whether the penetration of E. tenella into such a lymphoid cell is a beneficial step for the parasite survival and development. Three sequential experiments were performed. First, the in vitro established bovine kidney cell line, MDBK cells, were evaluated for use as host cells for E. tenella, through morphological observation. Second, the degree of parasite development and multiplication in MDBK cells was quantitatively assayed using radioisotope-labelled uracil (3H-uracil). Third, the E. tenella sporozoites viability was assayed after preincubation of them with chicken spleen cells. E. tenella o?cysts obtained from the ceca of the infected chickens were used for the source of the sporozoites. Spleen cells (E) obtained from normal chickens (FP strain) were preincubated with the sporozoites (T) at the E:T ratio of 100:1, 50:1 or 25:1 for 4 or 12 hours, and then the mixture was inoculated into the MDBK cell monolayer. Morphologically the infected MDBK cells revealed active schizogonic cycle of E. tenella in 3-4 days, which was characterized by the appearance of trophozoites, and immature and mature schizonts containing merozoites. The 3H-uracil uptake by E. tenella increased gradually in the MDBK cells, which made a plateau after 48-60 hours, and decreased thereafter. The uptake amount of 3H-uracil depended not only upon the inoculum size of the sporozoites but also on the degree of time delay (preincubation; sporozoites only) from excystation to inoculation into MDBK cells. The 3H-uracil uptake became lower as the preincubation time was prolonged. In comparison, after preincubation of sporozoites with spleen cells for 4 or 12 hours, the 3H-uracil uptake was significantly increased compared with that of control group. From the results, it was inferred that, although the penetration of E. tenella sporozoites into the lymphoid cells such as IEL is not an essential step, it should be at least a beneficial one for the survival and development of sporozoites in the chicken intestine.     

Ultrastructural observations on the developmental stages of Lankesterella minima (Apicomplexa) in experimentally infected Rana catesbeiana tadpoles

During May to August 1988, the prevalence of Lankesterella minima in bullfrog tadpoles and adults in the vicinity of Lake Sasajewun, Algonquin Park, Ontario, was 54.8% and 29.4%, respectively. The parasite was transmitted to laboratory-reared tadpoles through the bite of experimentally infected, laboratory-reared leeches (Batracobdella picta). Intraerythrocytic sporozoites were observed in the experimentally infected tadpoles 36 days postexposure (PE) to leech bites. Sporogonic stages in these tadpoles were examined by light and electron microscopy. Oocysts developed in vascular endothelial cells of several organs. Multinucleate oocysts at 29 days PE lay in a parasitophorous vacuole and contained several large inclusions. Mature oocysts, observed 32 days PE, contained about 70 sporozoites that exhibited typical apicomplexan features. In the posterior region of the sporozoites, 12 helically arranged chains of electron-lucent intrapellicular extended anteriorly from a dense terminal ring.     

Observations on the leech Placobdella ornata feeding from bony tissues of turtles

The leech Placobdella ornata was observed feeding from the blood sinuses of the plastron and carapace bones of Chelydra serpentina and Chrysemys picta. Evidence of successful feeding included blood upwelling from the point of attachment and gastric ceca of the leeches freshly filled with blood after removal. There was an apparent preference for the sulci between scales of the shell.     

Inhibition of Ca2+/calmodulin-dependent protein kinase blocks morphological differentiation of plasmodium gallinaceum zygotes to ookinetes

Once ingested by mosquitoes, malaria parasites undergo complex cellular changes. These include zygote formation, transformation of zygote to ookinete, and differentiation from ookinete to oocyst. Within the oocyst, the parasite multiplies into numerous sporozoites. Modulators of intracellular calcium homeostasis, MAPTAM, and TMB-8 blocked ookinete development as did the calmodulin (CaM) antagonists W-7 and calmidazolium. Ca(2+)/CaM-dependent protein kinase inhibitor KN-93 also blocked zygote elongation, while its ineffective analog KN-92 did not have such effect. In vitro both zygote and ookinete extracts efficiently phosphorylated autocamtide-2, a classic CaM kinase substrate, which could be blocked by calmodulin antagonists W-7 and calmidazolium and CaM kinase inhibitor KN-93. These results demonstrated the presence of calmodulin-dependent CaM kinase activity in the parasite. KN-93-treated parasites, however, expressed the ookinete-specific enzyme chitinase and the ookinete surface antigen Pgs28 normally, suggesting that the morphologically untransformed parasites are biochemically mature ookinetes. In mosquitoes, KN-93-treated parasites did not develop as oocysts, while KN-92-treated parasites produced similar numbers of oocysts as controls. These data suggested that in Plasmodium gallinaceum morphological development of zygote to ookinete, but not its biochemical maturation, relies on Ca(2+)/CaM-dependent protein kinase activity and demonstrated that the morphological differentiation is essential for the further development of the parasite in infected blood-fed mosquitoes.