猪尾猴疟原虫广西株红前期的研究

作者观察了猪尾猴疟原虫广西株在熊猴和猕猴肝内第6、7、8天的红前期裂殖体。红前期裂殖体为椭圆形,较小,成熟裂殖体不超过30微米,含裂殖子平均约1500个。在发育过程中突出的特征是有细胞岛(Pseudocytomere)形成,完成裂体增殖时间仅8—9天。本虫株在形态及发育方面与猪尾猴疟原虫的OS株极为相似。     

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。     

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。     

猪肾虫(Stephanurus dentatus Diesing,1839)的发育和移行的研究

1.猪肾虫从口吞食和皮肤接触均可感染,小猪皮肤接触和吞食感染相同,大猪皮肤较厚,幼虫不易侵入,其获得感染是以口吞食为主。 2.小猪从皮肤接触感染,经3—5天在皮下结缔组织行第三次蜕皮,38天移行至肝,150天虫体穿出肝表面向腹腔柔组织移行,157—185天在输尿管壁上形成包囊发育为成虫。从口吞食感染,经3—4天在胃壁大网膜和肠系膜等处停留,形成结节病变行第三次蜕皮,25天移行至肝,30天雌雄已明显分化,40日前后行弟四次蜕皮,118天离开肝向腹腔移行,156天后在输尿管壁形成包囊发育为成虫。皮下注射感染与皮肤接触感染相同。血管注射感染,幼虫到肺后多数不发育而死亡。 3.猪肾虫侵入宿主后,有嗜组织的习性,在组织中停留和移行,移行的方向不定,可向体前部移行,亦可向体后部移行。在淋巴结中常检得虫体,门静脉中虽有虫体栓塞,但未见在血管和淋巴管中移行。 4.猪肾虫在宿主体中发育的时间,除因虫体停留在组织结节中长短不定外,移行的速度与宿主大小有关系;宿主小,移行较速,成熟较快;宿主体大,移行时间较长,成熟较慢;因此致使过去学者报告猪肾虫成熟时间有4个月,5个月,6个月和9个月等不同结论。 5.从豚鼠皮肤接触感染,经3—5天幼虫皮下结缔组织,肠系膜和大网膜等柔组织形成结节病变,行第三次蜕皮,经5—8天移行至     

环形泰勒焦虫裂殖体细胞培养及其免疫原性的研究

1.本实验中所试用的BLS和“乳叶”培养液,都能良好地支持环形泰勒焦虫裂殖体感染的成淋巴细胞贴壁和悬浮生长,在4天内细胞量增殖5倍以上,传代以后的成淋巴细胞95%以上都寄生有裂殖体。成淋巴细胞“乳叶”培养液中,已连续传代培养15个月以上,仍生长良好,初步证明某些培养液中所加入的多种维生素等成份,对促进寄生有裂殖体的成淋巴细胞的生长繁殖,不是必要的。 2.传代细胞内的裂殖体,仅见无性裂殖体一种。裂殖体的分裂方式与宿主细胞的有丝分裂是密切相关的,在成淋巴细胞分裂的后期,裂殖体核沿纺锤丝排列成一行或数行,然后几乎被均等地分配到两个子细胞中去。裂殖体是细胞分裂的一种强烈刺激物。 3.成淋巴细胞在明胶制剂中,于4—6℃下保存到40—50天仍不失其再生长繁殖的能力。 4.裂殖体具有坚强的免疫原性,接种牛体后可产生抗环形泰勒焦虫病的免疫力。传代培养45天,2个半月和5个半月的细胞接种牛体后,前二种细胞形成带虫免疫,后者为非带虫免疫,安全性皆表现良好。经蜱叮咬攻毒,17头牛100%得到保护。516×10~5—5×10~6细胞剂量均安全有效。用明胶制剂作为保护剂,所制的胶冻细胞苗在4—6℃下保存20天和30天,对牛体仍然可产生坚强的保护能力,即疫苗的有效保存期可达30天。     

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。     

体外培养牛白细胞中环形泰勒焦虫(Theileria annulata)的增殖、释放与感染

在体外培养的牛外周血白细胞中,环形泰勒焦虫裂殖子与裂殖体寄生于宿主细胞的细胞质中,并且随着宿主细胞的分裂而分到两个子细胞中。焦虫染色质粒的分裂方式为二分裂,随着焦虫颗粒的不断增殖,逐渐发育为成熟的裂殖体。体外培养感染焦虫的牛白细胞可通过伪足与细胞裂解两种途径向培养液中释放焦虫颗粒。释放到培养液中的焦虫颗粒对体外培养的健康牛外周血白细胞具有感染能力,感染细胞能在体外连续传代培养。     

广东肝血簇虫在宿主内脏裂体生殖时期超微结构的研究

本文详细描述了广东肝血簇虫(Hepatozoon guangdongensis)在实验宿主、中国水蛇肺部裂体生殖整个发育过程各期虫体的超微结构。成熟裂殖体内的裂殖子与肺部毛细血管内皮细胞内的裂殖子的超微结构是相似的。裂殖子(3.4×1.3μm)外被由外膜和内膜构成的表膜,它与球虫一样具有包括类锥体在内的完全顶复结构。内皮细胞内的裂殖子和滋养体都没有围虫泡和围虫泡膜包绕。具有内膜的长形滋养体变圆,并外被由宿主细胞产生的围虫泡和围虫泡膜包绕,转变成为圆形的幼期裂殖体,然后发育成为成熟的裂殖体。成熟裂殖体(23×10μm)内含30—50个裂殖子。裂殖体内没有观察到残余体存在。     

广东肝血簇虫(Apicomplexa:Haemogre-garinidae)感染试验和生活史的研究

作者利用致乏库蚊(Culex fatigans)作为实验媒介,把寄生在灰鼠蛇(Ptyas korros)的广东肝血簇虫(Hepatozoon guangdongensis)感染到无虫的灰鼠蛇(渔游蛇(Natrix piscator)、三索锦蛇(Elaphe radita)和中国水蛇(Enhydris chinesis);观察其生活史。首次描述了由x裂殖体产生的小裂殖子,进入红细胞,经滋养体和幼小配子母体阶段发育成配子母体的过程,以及由y裂殖体产生的大裂殖子继续在内脏进行无性生殖的细节;在对广东肝血簇虫生活史进行研究的基础上,提出一种新的肝血簇虫在脊椎动物体内发育的模式和血液时期各期虫体划分的形态学标准。     

海南岛蚋科昆虫的初步调查

１９９３年３—７月,在海南岛三亚、尖峰岭和通什地区进行了蚋科昆虫的调查。经鉴定为黄足真蚋Ｓｉｍｕｌｉｕｍ（Ｅｕｓｉｍｕｌｉｕｍ）ａｕｒｅｏｈｉｒｔｕｍＢｒｕｎｅｔｔｉ,１９１１、查头真蚋Ｓ．（Ｅ．）ｃｈｉｔｏｅｎｓｅＴａｋａｏｋａ,１９７９、节蚋Ｓ．（Ｓｉｍｕｌｉｕｍ）ｎｏｄｏｓｕｍＰｕｒｉ,１９１１、五条蚋Ｓ．（Ｓ．）ｑｕｉｎｑｕｅｓｔｒｉａｔｕｍ（Ｓｈｉｒａｋｉ）,１９３５、亮胸蚋Ｓ．（Ｓ．）ｎｉｔｉｄｉｔｈｏｒａｘＰｕｒｉ,１９３２、海南绳蚋Ｓ．（Ｇｏｍｐｈｏｓｔｉｌｂｉａ）ｈｉａｎａｎｅｎｓｉｓｌｏｎｇｅｔＡｎ,１９９４、尖峰绳蚋Ｓ．（Ｇ．）ＪｉａｎｆｅｎｇｅｎｓｉｓＡｎｅｔＬｏｎｇ,１９９４、后宽绳蚋Ｓ．（Ｇ．）ｍｅｔａｔａｒｓａｌｅＢｒｕｎｅｔｔｉ,１９１１、怕头绳蚋Ｓ．（Ｇ．）ｐａｔｔｏｎｉＳｅｎｉｏｒｙｉ－Ｗｈｉｔｅ,１９９２（中国新纪录）和Ｓ．（Ｓ．）ｓｐ,共１属３亚属１０种蚋。其中黄足真蚋、五条蚋和节蚋为常见蚋种。     

Survival and Development of Eimeria meleagrimitis Tyzzer, 1929 in Bovine Kidney and Turkey Intestine Cell Cultures

SYNOPSIS. Monolayer cell cultures of embryonic turkey intestine (primary) and bovine kidney (cell line, 20th passage), maintained at 40.6 and 43 C for alternating intervals of approximately 12 hours in Basal Medium Eagle and fetal calf serum at pH 7.0–7.4, were observed for 144 hours after inoculation of Eimeria meleagrimitis sporozoites.
In turkey intestine cultures, which consisted of fibroblast-like cells and patches of epitheliul-like cells, there were decreases of 80 and 81% in the numbers of parasites between 5 and 48 hrs; in bovine cultures, 21–41% decreases. Decreases in the turkey cultures, however, were due to the nonsurvival of sporozoites in fibroblast-like cells; in epitheliul-like cells there was a 42% dcrease between 5 and 48 hrs and only 27% between 48 and 144 hours.
Trophozoites were present in bovine cells at 5 hrs. Small, mature schizonts containing only 12-28 merozoites were present in the bovine cultures and in the epitheliul-like cells within turkey intestine cultures from 48-144 hrs. Larger schizonts (50-115 by 20-70 μ) were present in bovine but not in turkey cultures from 72–144 hrs. Many of these schizonts contained far more merozoites than schizonts of any of the 3 generations described from the host.
In bovine cultures, there was an abundance of liberated merozoites at 50, 52, 74, and 76 hrs; many had reinvaded cells, sometimes as many as 50–60 per cell. In turkey cultures, liberated merozoites were found once at 144 hrs and none were intracellular. At 120 and 144 hrs in bovine cultures, abnormally developed and degenerate forms appeared; in turkey cultures, all were normal.     

Development of Eimeria meleagrimitis Tyzzer from sporozoites and merozoites in turkey kidney cell cultures

Sporozoites and 1st-, 2nd-, and 3rd-generation merozoites of Eimeria meleagrimitis were inoculated into primary cultures of turkey kidney cells. In vitro-excysted sporozoites developed into mature macrogamonts in 8 days; in vivo-excysted sporozoites developed into 2nd- or 3rd-generation schizonts within 5 to 7 days. First-generation merozoites obtained from infected turkeys produced mature 2nd-generation schizonts within 24 h. Second-generation merozoites from turkeys produced mature macrogamonts and oocysts within 72 h, whereas 3rd-generation merozoites produced these stages within 48 h. The oocysts that developed from 3rd-generation merozoites sporulated at 25 C and were infective for turkeys. The timing of the early stages and the intervals between schizogonic generations in cultures were comparable with those in turkeys. Morphologic parameters, however, indicated that some differences existed between in vitro and in vivo development. Second- and 3rd-generation schizonts and gamonts that developed after inoculation of cultures with merozoites were similar to stages in turkeys. Oocysts, however, were significantly smaller (P less than 0.05) in cultures. All stages that developed after inoculation of cultures with sporozoites were smaller (P less than 0.05) than their in vivo counter parts.     

Development of Eimeria meleagrimitis Tyzzer from Sporozoites and Merozoites in Turkey Kidney Cell Cultures*

SYNOPSIS. Sporozoites and 1st-, 2nd-, and 3rd-generation merozoites of Eimeria meleagrimitis were inoculated into primary cultures of turkey kidney cells. In vitro-excysted sporozoites developed into mature macrogamonts in 8 days; in vivo-excysted sporozoites developed into 2nd- or 3rd-generation schizonts within 5 to 7 days. First-generation merozoites obtained from infected turkeys produced mature 2nd-generation schizonts within 24 h. Second-generation merozoites from turkeys produced mature macrogamonts and oocysts within 72 h, whereas 3rd-generation merozoites produced these stages within 48 h. The oocysts that developed from 3rd-generation merozoites sporulated at 25 C and were infective for turkeys. The timing of the early stages and the intervals between schizogonic generations in cultures were comparable with those in turkeys. Morphologic parameters, however, indicated that some differences existed between in vitro and in vivo development. Second- and 3rd-generation schizonts and gamonts that developed after inoculation of cultures with merozoites were similar to stages in turkeys. Oocysts, however, were significantly smaller (P < 0.05) in cultures. All stages that developed after inoculation of cultures with sporozoites were smaller (P < 0.05) than their in vivo counter parts.     

Eimeria tenella: use of a monoclonal antibody in determining the intracellular fate of the refractile body organelles and the effect on in vitro development

A monoclonal antibody, which recognizes the refractile body of Eimeria sporozoites, was used to study the developmental fate of this organelle during asexual development of E. tenella and to determine the effect of this monoclonal antibody on in vitro development of the parasite. Through use of immunofluorescent antibody and gold-labeling techniques at the light and electron microscopy level, the refractile body at 48 to 96 hr postinoculation was found to separate into 6 to 10 small globules, then diffuse throughout the schizont cytoplasm, and eventually reconcentrate as a small dot of material in each of the mature first-generation merozoites. The schizont did not develop to maturity if diffusion of the refractile body did not occur. The refractile body material was quickly lost as the merozoite left the schizont and invaded new cells and was not detected in any later developmental stages. The in vitro development of first- and second-generation schizonts of E. tenella was greatly inhibited (up to 100%) with exposure to the monoclonal antibody. There was an increase in the number of schizonts with nondispersed refractile body in the monoclonal antibody-treated cells when compared to the untreated controls, and the few mature schizonts seen had up to a 50-fold decrease in the number of merozoites. Immunofluorescent antibody labeling of the refractile body of intracellular sporozoites and schizonts treated in vitro with the monoclonal antibody for 24-96 hr postinoculation indicated that the antibody had crossed the host cell and parasite plasma membrane during incubation.     

约氏疟原虫红外期成熟裂殖体的超微结构研究

用细胞色素氧化酶组织化学方法处理感染了约工疟原虫子孢子的大鼠肝脏,通过透射电镜研究红外期裂殖体的超微结构。在接种子孢子后４８小时的标本中发现一成熟裂殖体,外周仍由一寄生虫质膜包裹,膜下有许多小泡,粗面内质肉、圆形或蚕豆形具明显嵴的线粒体,以及大量成熟裂殖子。     

Schizogonic Development of Leucocytozoon smithi

ABSTRACT The schizogonic development of Leucocytozoon smithi in the liver of experimentally infected turkey poults was examined by electron microscopy. Following intraperitoneal injection, sporozoites migrated to the liver and entered hepatic cells to become intracellular trophozoites. Three to four days post inoculation (PI), trophozoites underwent asexual multiple fission known as merogony or schizogony. Two generations of schizonts were observed. The primary or first generation schizonts, abundant on day 4 PI, appeared as interconnected cytoplasmic masses (pseudocytomeres). Each pseudocytomere was enclosed by a membranous vacuole and contained varying numbers of nuclei. As nuclear division and growth of the schizonts continued, larger discrete cytoplasmic masses or cytomeres were formed with rhoptries and multiple nuclei in various stages of division. Synchronous multiple cytoplasmic cleavage of the schizont resulted in the formation of numerous uninucleate merozoites. Second generation schizonts, which developed from hepatic merozoites released from primary schizonts, were abundant in hepatocytes on day 6 PI. Although tissue samples from liver, lung, spleen, kidney, intestine, brain, blood vessels and lymph nodes were examined, schizogonous forms were observed in liver only. No megaloschizonts were detected in any host tissue examined. Schizogonic development was completed by day 7 PI as merozoites developed into gametocytes within mononuclear phagocytes.     

Hepatozoon ursi n. sp. (Apicomplexa: Hepatozoidae) in Japanese black bear (Ursus thibetanus japonicus)

Morphological and genetic features of a new Hepatozoon species, Hepatozoon ursi n. sp., in Japanese black bear (Ursus thibetanus japonicus) were studied. Schizogonic developmental stages were observed in the lungs of Japanese black bears. The schizonts were sub-spherical in shape and 45.7+/-4.6 x 42.7+/-4.5 microm in size. Each mature schizont contained approximately 80-130 merozoites and 0-5 residual bodies. The merozoites were 7.0+/-0.7 x 1.8+/-0.3 microm in size. Intraleukocytic gametocytes were slightly curved, cigar-like in shape and had a beak-like protrusion at one end. The size of the gametocytes was 10.9+/-0.3 x 3.3+/-0.2 microm. The analyses of the18S rRNA gene sequences supported the hypothesis that H. ursi n. sp. is different from other Hepatozoon species. Mature Hepatozoon oocysts were detected in two species of ticks (Haemaphysalis japonica and Haemaphysalis flava) collected on the bears infected with H. ursi n. sp. Two measured oocysts were 263.2 x 234.0 microm and 331.8 x 231.7 microm, respectively. The oocysts contained approximately 40 and 50 sporocysts, respectively. The sporocysts were sub-spherical in shape and 31.2+/-2.5 x 27.0+/-2.9 microm in size. Each sporocyst contained at least 8-16 sporozoites, with the sporozoites being 12.2+/-1.4 x 3.5+/-0.5 microm in size. H. ursi n. sp. is the first Hepatozoon species recorded from the family Ursidae.     

Cultivation of Eimeria tenella in Japanese quail embryos (Coturnix coturnix japonica).

Complete development of Eimeria tenella in Japanese quail embryos was observed. Sporozoites were inoculated into the allantoic cavity of 7-day-old Japanese quail embryos (Coturnix coturnix japonica), after which the infected embryos were incubated at 41 C. In the chorioallantoic membrane mature first generation schizonts, mature second generation schizonts, and gametes were detected at 48 hr postinoculation of sporozoites (PI), 84 hr PI, and 126 hr PI, respectively. Mature gametes and zygotes were found at 132 hr PI, and oocysts were detected at 138 hr PI. Mortality of embryos increased with increment of inoculum size of sporozoites. LD50 was 1.7 x 10(2) sporozoites. Oocyst production was also dependent on inoculum size. Oocysts harvested from embryos sporulated. The oocysts were inoculated into 13-day-old chickens, and oocysts, capable of sporulating normally, were recovered from ceca 7 days after inoculation.     

Development of Eimeria callospermophili and E. bilamellata from the Uinta Ground Squirrel Spermophilus armatus in Cultured Cells*

SYNOPSIS. Cell lines or established cell lines of bovine, ovine or human origin and primary cells from whole embryos of groundsquirrels were used in a study of the in vitro development of Eimeria callospermophili and E. bilamellata from the Uinta ground squirrel, Spermophilus armatus. Monolayers in Leighton tube cultures were inoculated with sporozoites of either of these 2 species and examined with phase-contrast microscopy at various intervals. After such examination, coverslips were fixed in Schaudinn's or Zenker's fluid and variously stained. E. callospermophi sporozoites penetrated cells and underwent development to mature 1st generation schizonts in most cell types. At different times after inoculation, both species formed sporozoite-shaped schizonts, which later became spheroidal. Intracellular movements of sporo zoite-shaped schizonts of E. callospermophili were observed and such schizonts penetrated cells when freed by mechanical disintegration of the host cells. Merozoites were formed at the periphery of the schizont in both species. Mature 1st generation schizonts of E. callospermophili, with 6–14 merozoites, were first seen 15 hr after inoculation; the corresponding values for E. bilamellata were 12–27 merozoites and 4 days. Merozoites of both had anterior and posterior refractile bodies. Exposure to a trypsin-bile solution stimulated motility in merozoites of E. callospermophili. Second generation trophozoites and immature schizonts of E. callospermophili were seen in cultures of primary cells of whole ground-squirrel embryos 20–24 hr and 44–48 hr, respectively, after inoculation of sporozoites.