中华硬蜱、越原血蜱和青海血蜱经期传播莱姆病螺旋体的实验研究

以实验室培育的非感染中华硬蜱、越原血蜱和青海血蜱刺叮人工感染21天的阳性KM鼠,利用分离培养和PCR方法检测蜱对莱姆病螺旋体Borrelia garinii的保持能力以及体内螺旋体对敏感动物的感染能力。中华硬蜱、越原血蜱和青海血蜱非感染幼蜱均可以通过吸血获得莱姆病螺旋体, 然而,它们在饱血、消化、蜕皮和再次吸血过程中对莱姆病螺旋体的保持能力差异很大。只有中华硬蜱可以保持活的莱姆病螺旋体直至蜕化为下一发育阶段,蜕化后仍携带莱姆病螺旋体并对敏感KM鼠有感染能力。而越原血蜱和青海血蜱对莱姆病螺旋体的保持能力不能跨越消化、蜕皮阶段,蜕化后这两种血蜱不再携带莱姆病螺旋体,因此,这两种血蜱不具备经期传播能力,作为莱姆病媒介的可能性不大。中华硬蜱于幼蜱到若蜱以及若蜱到成蜱两个阶段均具备经期传播莱姆病螺旋体的能力,提示了中华硬蜱作为南方莱姆病的潜在媒介是可能的。     

精河株斑点热立克次体在非洲钝缘蜱体内的分布

本文报告用乙二醇甲基丙烯酸(GMA)包埋、切片和姬姆萨染色观察精河株斑点热立克次体在非洲钝缘蜱(Ornithodoros moubata)体内的分布情况。接种立克次体后2天,即可从蜱的血淋巴涂片中查见立克次体,但各器官和组织的切片6天后才开始出现阳性,尤以中肠最迟。叮咬感染的蜱的中肠感染较早也较严重。无论接种或叮咬感染,除睾丸和精子外,所查的器官和组织包括涎腺、基节器、中肠、直肠、卵巢以及雄蜱的附腺和贮精囊等切片中都查见立克次体。有的蜱感染很严重,立克次体充满了许多器官和组织的细胞。     

中华硬蜱和二棘血蜱的交叉免疫反应

本文首次比较了经中华硬蜱(Ixodes sinensis)叮咬三次后再经二棘血蜱(Haemaphysalisbispinosa)叮咬的家兔与仅经二棘血蜱叮咬的家兔的交叉免疫抗性。二棘血蜱叮咬被中华硬蜱致敏的家兔时,吸血增重为:143.12±32.67mg,但二棘血蜱在正常家兔体上寄生,初次吸血增重为:181.30±44.35mg,两者之间有显著性差异(P<0.01)。中华硬蜱和二棘血蜱唾液腺提取物(SGE)经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),显示两者分别有24条和22条电泳带,中华硬蜱主带有6条,分子量分别为142、105、94、66/65、64和56kD,而二棘血蜱主带有5条,分子量分别为:215、114、105、66/65和58kD,经中华硬蜱叮咬致敏的家兔血清和经二棘血蜱叮咬致敏的家兔血清作免疫印渍,均显示出105kD这一电泳带。该实验表明中华硬蜱和二棘血蜱叮咬家兔两者之间存在着交叉免疫反应,提示105kD蛋白质抗原可能是两者的共同抗原。     

银盾革蜱的中肠上皮变化与血餐消化

用光学显微镜及电子显微镜对不同生理状况下银盾革蜱Dermacentor niveus Neumann雌虫中肠上皮及血餐消化进行了研究.饿蜱的中肠只由一种干细胞组成,脂滴作为饿蜱营养的贮藏形式.非滞育蜱消化分三个阶段,即第一连续消化阶段,减慢消化阶段及第二连续消化阶段.吸血后中肠上皮共观察到四种细胞类型,即替代细胞、分泌细胞、消化细胞及卵赞原细胞.滞育蜱第一连续消化阶段延长.饱血后60天到120天,消化作用几近停止,为停滞消化阶段.卵黄原细胞的超微结构有明显改变.滞育解除后,开始进行减慢消化阶段及第二连续消化阶段.     

华硬蜱和二棘血蜱的交叉免疫反应

本文首次比较了经中华硬蜱(Ixodes sinensis)叮咬三次后再经二棘血蜱(Haimaphysalis bispinosa)叮咬的家兔与仅经二棘血蜱叮咬的家兔的交叉免疫抗性。二棘血蜱叮咬被中华硬蜱致敏的家兔时,吸血增重为：143.12±32.67mg,但二棘血蜱在正常家兔体上寄生,初次吸血增重为：181.30±44.35mg,两者之间有显著性差异(P<0.01)。中华硬蜱和二棘血蜱唾液腺提取物(SGE)经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),显示两者分别有24条和22条电泳带,中华硬蜱主带有6条,分子量分别为142、105、94、66/65、64和56kD,而二棘血蜱主带有5条,分子量分别为：215、114、105、66/65和58kn,经中华硬蜱叮咬致敏的家兔血清和经二棘血蜱叫‘咬致敏的家兔血清作免疫印渍,均显示出105kD这一电泳带。该实验表明中华硬蜱和二棘血蜱叮咬家兔两者之间存在着交叉免疫反应,提示105kD蛋白质抗原可能是两者的共同抗原。     

苦皮藤素V对东方粘虫中肠细胞及其消化酶活性的影响

苦皮藤素V是从杀虫植物苦皮藤Celastrus angulatus Max.根皮中分离的一种对昆虫具有毒杀活性的新化合物。该文通过电镜观察和生化分析研究了其对东方粘虫Mythimnaseparata(walker)幼虫中肠组织及中肠主要消化酶活性的影响。电镜观察发现,中毒试虫的中肠细胞及其细胞器发生明显病变：柱状细胞顶膜微绒毛零乱、减少;线粒体肿胀,出现空白亮区,双层膜不完整;细胞质密度降低,细胞器排列紊乱;内质网池扩张,囊泡化,粗面内质网减少;杯状细胞杯腔变大,微绒毛减少。消化酶活性测定结果表明,中毒试虫中肠的蛋白酶、淀粉酶及脂肪酶的活性和正常虫相比,无显著变化。因此认为,苦皮藤素V主要作用于中肠细胞的质膜及其内膜系统。     

取食转Bt基因抗虫玉米后亚洲玉米螟幼虫中肠的组织病理变化

利用透射显微镜（TEM）观察亚洲玉米螟Ostrinia furnacalis (Guenée)幼虫取食了表达Cry1Ab杀虫蛋白的转Bt基因玉米心叶组织后中肠的组织病理变化, 以探讨转Bt基因玉米对亚洲玉米螟的致病机理, 为其合理、安全和持续利用提供理论依据。结果表明：亚洲玉米螟取食Bt玉米后中肠细胞及其细胞器发生了明显的病变。取食Bt玉米12 h后中肠细胞开始病变, 首先微绒毛脱落、内质网开始肿胀, 24 h后内质网肿胀、增多, 杯状细胞杯腔增大, 48 h后微绒毛大量脱落, 细胞开始空泡化, 随着取食时间的增加, 细胞空泡化程度加剧, 在感染前期细胞间的病变程度差异较大。微绒毛脱落、内质网肿胀断裂是在多数取食Bt玉米的亚洲玉米螟中肠细胞发生的普遍病变。由此表明, 人工修饰的Cry1Ab基因导入到玉米染色体组中所表达的杀虫蛋白可使玉米螟幼虫中肠细胞发生病变, 最终导致其死亡。     

苏芸金杆菌感染粘虫后中肠组织学病变的研究

本文描述了Bacillus thuringiensis HD-1纯晶体感染粘虫Mythimna separata 5龄初幼虫后,其中肠肠壁细胞的超微结构变化.电镜观察表明:首先线粒体,发生病变,随后,细胞顶部肿胀、胞质电子密度下降,顶部胞质之下的各种细胞器均受损伤.随着病变加重,柱状细胞基膜内褶及杯状细胞质突起,这两个富含线粒体的区域发生病变最为明显.最终,肠壁细胞解体从基膜脱落.     

单宁酸体外抑制痘苗病毒感染活性的效果分析

目的:观察单宁酸体外抑制痘苗病毒(VACV)感染的效果,探讨单宁酸干扰病毒入侵宿主细胞膜脂的效应机制。方法:基于痘苗病毒细胞感染模型,在病毒吸附前、吸附后和吸附同时3种情况下分别添加单宁酸,采用中性红比色法检测单宁酸对VACV的抑制作用,利用荧光偏振法和流式细胞术分别测定VACV对细胞膜流动性和膜电位的影响及单宁酸的干预作用。结果:单宁酸以病毒吸附后及病毒吸附同时2种方式加入时可抑制VACV感染,半数抑制浓度(IC50)分别为8.7和13.1μg/m L,治疗指数(TI)分别为9.3和6.2;VACV感染可增加细胞膜流动性,单宁酸在病毒吸附后作用,可显著降低细胞膜流动性;VACV感染使细胞膜电位绝对值增大,单宁酸对VACV感染引起的细胞膜超极化状态没有明显影响。结论:单宁酸具有明显的体外抑制VACV感染宿主细胞的作用,其机制可能与拮抗因病毒感染所致的细胞膜流动性升高的膜脂干扰效应有关。     

茶毛虫核多角体病毒侵染后宿主细胞学的病变

本文应用超薄切片、冰冻断裂、扫描电镜等技术,对茶毛虫核多角体病毒侵染后宿主细胞病理超微结构的变化等进行了研究。结果表明:病毒侵染72—120小时后,宿主中肠细胞器,如线粒体、内质网、核糖体、板层小体、溶酶体等均有明显的病变。同时,在中肠细胞表面有病毒粒子吸附外,细胞表面的微绒毛亦有倒塌肿胀等病症表现。     

Echinococcus granulosus: ultrastructure of epithelial changes during the first 8 days of metacestode development in vitro

The epithelium of artificially hatched and activated oncospheres of E. granulosus was studied ultrastructurally over the first 8 days of metacestode development in vitro. Within 4 h of activation, the epithelium was transformed from a thin cytoplasmic layer into a much wider layer packed with penetration gland granules and containing mitochondria and Golgi apparatus. Microvilli were extended from the outer plasma membrane and the basal lamina on the inner epithelial surface virtually disappeared. Microvilli increased in number and length over the first 24 h of development while granules in both the epithelium and penetration gland decreased in number. The granules appear to be involved in microvilli formation. After 3 days of development, the first lamination resolved ultrastructurally as shortened microvilli and some microtriches extending from the epithelium surrounded by an electron-dense microfibrillate material containing sloughed microvilli. By 6 days post-activation, no microvilli remained and only double-walled truncated microtriches extended from the epithelium. The microfibrillate material had become more electron-dense and was closer to the epithelium than at day 1. Within 8 days of metacestode development, a second lamination had developed. Both microfibrillate and particulate material of a greater electron density than the first lamination was added to the microthrix side of the first lamination.     

Ultrastructure of midgut endocrine cells in the adult mosquito, Aedes aegypti

The ultrastructure of endocrine cells in the midgut of the adult mosquito, Aedes aegypti, resembled that of endocrine cells in the vertebrate gastro-intestinal tract. Midgut endocrine cells, positioned basally in the epithelium as single cells, were cone-shaped and smaller than the columnar digestive cells. The most distinctive characteristic of endocrine cells was numerous round secretory granules along the lateral and basal plasma membranes where contents of the granules were released by exocytosis. Secretory granules in each individual cell were exclusively of one type, either solid or 'haloed', and for all cells observed, the range in granule diameter was 60-120 nm. The cytoplasm varied in density from clear to dark. Lamellar bodies were prominent in the apical and lateral cellular regions and did not exhibit acid phosphatase activity. The basal plasma membrane was smooth adjacent to the basal lamina, whereas in digestive cells the membrane formed a labyrinth. Some endocrine cells reached the midgut lumen and were capped by microvilli; a system of vesicles and tubules extended from beneath the microvilli to the cell body. An estimated 500 endocrine cells were distributed in both the thoracic and abdominal regions of the adult midgut. In one midgut, we classified a sample of endocrine cells according to cytoplasmic density and granule type and size; endocrine cells with certain types of granules had specific distributions within the midgut.     

Digestive cells in the midgut of Triatoma vitticeps (Stal, 1859) in different starvation periods

Triatoma vitticeps (Stal, 1859) is a hematophagous Hemiptera that, although being considered wild, can be found in households, being a potential Chagas’ disease vector. This work describes the histology and ultrastructure of the midgut of T. vitticeps under different starvation periods. Fifteen adults of both sexes starved for 3, 7, 20 and 25 days were studied. In general, digestive cells had apical microvilli, basal plasma membrane infoldings and central nucleus. The perimicrovillar membrane was found in all insects examined. Digestive cells of anterior midgut had lipid droplets, glycogen granules, developed basal labyrinth associated with mitochondria suggesting their role in nutrient storage and in fluid and ion transport. The cells of median and posterior regions of the midgut were rich in rough endoplasmic reticulum, lysosomes, vesicles and granules with different electron-densities. Moreover, cells of the posterior portion of the midgut had hemozoyn granules and mitochondria in the apical cytoplasm close to microvilli, suggesting their role in blood digestion and active nutrient absorption. The midgut of T. vitticeps showed differences in digestive cells associated with the time after feeding, and the increase of vesicles amount in long starvation periods, which suggests enzyme storage, which is readily used after a blood meal.     

Fine structure of the midgut of Sinopanorpa tincta (Navás) (Mecoptera: Panorpidae)

Fine structure of the midgut and degeneration of the midgut epithelium of the scorpionfly Sinopanorpa tincta (Navás) adults were investigated using light microscopy and scanning and transmission electron microscopy. The results show that the tubular midgut lacks gastric caeca and is composed of an outer longitudinal and an inner circular muscle layer, a basal lamina, an epithelium and a lumen from the outside to inside. A peritrophic membrane was not found in the lumen. A mass of nodules was observed on the surface of the basal lamina. Three types of cells were recognized in the epithelium: digestive, secretory, and regenerative cells. The digestive cells contain irregular-shaped infoldings in the basal membrane and two types of microvilli in the apical membrane. The secretory cells are characterized by irregular shape and large quantities of secretory granules in the basal cytoplasm. The regenerative cells are triangular in shape and distributed only in the nodules. The epithelial cells are degenerated through programmed cell-death mechanisms (apoptosis and necrosis). The type, function, and degeneration of the epithelial cells of the midgut are briefly discussed.     

Morphological and enzymatic analysis of the midgut of Anopheles darlingi during blood digestion

The midgut of adult female Anopheles darlingi is comprised of narrow anterior and dilated posterior regions, with a single layered epithelium composed by cuboidal digestive cells. Densely packed apical microvilli and an intricate basal labyrinth characterize each cell pole. Before blood feeding, apical cytoplasm contains numerous round granules and whorled profiles of rough endoplasmic reticulum. Engorgement causes a great distension of midgut. This provokes the flattening of digestive cells and their nuclei. Simultaneously, apical granules disappear, the whorls of endoplasmic reticulum disassemble and 3h post bloodmeal (PBM), nucleoli enlarge manyfold. An intense absorptive process takes place during the first 24 h PBM, with the formation of large glycogen inclusions, which persist after the end of the digestive process. Endoproteases activities are induced after bloodmeal and attain their maximum values between 10 and 36 h PBM. At least two different aminopeptidases seem to participate in the digestive process, with their maximum activity values at 36 and 48 h PBM, respectively. Coarse electrondense aggregates, possibly debris from digested erythrocytes, begin to appear on the luminal face of the peritrophic membrane from 18 h PBM and persist during all the digestive process, and are excreted at its end. We suggest that these aggregates could contain some kind of insoluble form of haem, in order of neutralize its toxicity.     

Functional ultrastructure of the midgut of the fire ant Solenopsis saevissima Forel 1904 (Formicidae: Myrmicinae)

Solenopsis saevissima has a midgut composed of columnar, regenerative, and goblet cells. The midgut epithelium was covered by a basal lamina. Outside the basal lamina, layers of inner oblique, circular, and outer longitudinal muscles were present. Columnar cells showed a basal plasma membrane containing numerous folds, mitochondria, and the nucleus. Rough endoplasmic reticulum, Golgi bodies, membrane bounded vacuoles, and spherocrystals were found in this region. The apical plasma membrane was constituted by microvilli, which were above a region rich in mitochondria. Regenerative cells were found in groups lying by the basal lamina. Goblet cells were associated with an ion-transporting mechanism between the haemolymph and the midgut epithelium. These cells were lying by the midgut lumen and large microvilli were evident, but the cytoplasmic features were similar to the columnar cells.     

Infection and replication sites of Spiroplasma kunkelii (Class: Mollicutes) in midgut and Malpighian tubules of the leafhopper Dalbulus maidis

Spiroplasma kunkelii distribution and infection mechanisms in the intestines and Malpighian tubules of Dalbulus maidis were investigated by transmission electron microscopy. Spiroplasmas were found between microvilli and in endocytic vesicles of the midgut epithelium. At the basal part, cytoplasmic vesicles contained multiple spiroplasmas with tube-like extensions and spiroplasmas accumulated between the laminae rara and densa of the basal lamina. Tip structures of flask-shaped spiroplasmas pierced the lamina densa that was discontinuous in close proximity to spiroplasmas. Spiroplasmas were found in hemolymph, crossed the basal lamina of Malpighian tubule epithelium and accumulated at high numbers in muscle cells that had cytopathogenic changes. S. kunkelii had perithrochous approximately 8nm diameter structures determined to be fimbriae protruding from the cell surface, and similar structures were adhering to the basal lamina of midgut epithelium and to external lamina of muscle cells. Further, spiroplasmas had pili-like appendages at one or both cell poles and appeared to conjugate. This is the first time that fimbriae and pili have been observed in a mollicutes.     

Ultrastructure of the encapsulation of Plasmodium cynomolgi (B strain) on the midgut of a refractory strain of Anopheles gambiae

Using transmission electron microscopy, we investigated the encapsulation of the simian malaria parasite, Plasmodium cynomolgi, in a refractory strain of the mosquito, Anopheles gambiae. After the ookinete penetrates the mosquito midgut epithelium and lodges between the basal membrane and the basal lamina, an electron-dense, melanin-like substance begins to coalesce around the parasite. Completely encapsulated parasites were found as early as 16 hr after the blood meal. Granules of the melanin-like substance often appeared to condense onto the parasite from the fluid in the extracellular spaces of the basal membrane labyrinth. Melanin granules also appeared to condense from the hemolymph onto the basal lamina underlying the parasite. In addition, groups of tubules, vesicles, and membranous whorls often were found in midgut cells that were located next to or were enclosing parasites. These structures were unusually electron-dense, and may have been associated with melanization. Hemocytes rarely were observed near completed capsules and neither hemocytes nor their remnants were components of the capsules. During later stages of encapsulation, parasites appeared abnormal and often were infiltrated with melanin. Although late-stage capsules were usually located basally, completed capsules enclosed by membranes were occasionally observed near the apical border of the midgut. Other capsules associated with cellular debris, were found in the lumen of the midgut from 1 to 6 days after the blood meal.     

庭疾灶螽中肠及马氏管结构

【目的】本研究旨在以庭疾灶螽Tachycines asynamorus为例探索驼螽消化系统和排泄系统在结构上与其生活环境的适应关系。【方法】运用解剖学方法、石蜡切片技术、冰冻切片技术及超薄切片技术对庭疾灶螽中肠及马氏管的结构进行研究。【结果】庭疾灶螽中肠向前延伸出3个胃盲囊包围着前胃。中肠上皮由再生细胞、柱状上皮细胞和内分泌细胞构成,具有典型的再生细胞龛;闭合型内分泌细胞紧贴在再生细胞龛的外围,基底区聚集大量的分泌颗粒。柱状上皮细胞内聚集有2类大的分泌颗粒：线团状颗粒和电子密度很高的球状颗粒;中肠管腔内有明显的围食膜结构,中肠基底部由基膜和肌肉层组成。马氏管着生在中后肠的交界处,从横切面看马氏管管壁具有3~5个细胞,细胞近管腔端部具有大量长微绒毛,细胞质内分布着电子致密的同心圆球晶体,基底膜内折形成膜迷路。【结论】庭疾灶螽中肠柱状上皮细胞的线团状颗粒由微丝包裹;内分泌细胞由再生细胞龛中的细胞分化而来,产生内分泌颗粒并将其排到血腔;中肠基膜发达,包含微丝与复合糖成分,基膜通过对中肠上皮细胞的支撑作用为肠道蠕动提供保障。庭疾灶螽马氏管细胞中可见大量颗粒和大量同心圆球晶体,推测可能是一种储存排泄。     

Sporogonic Development of Leucocytozoon smithi

The sporogonic development of Leucocytozoon smithi in its black fly vector was studied by light and electron microscopy and was compared with that of other haemosporidians. Within 18 to 24 h after ingestion of gametocytes by black flies, ookinetes passing through the midgut epithelium were observed. Intracellular migration of ookinetes resulted in the apparent disruption and degeneration of host cells. Intercellular migration also occurred as was evidenced by the presence of ookinetes between midgut cells. Transformation of ookinete to spherical oocyst occurred extracellularly in three different sites. Although most oocysts were found between the host cell basal membrane and the basal lamina, large numbers also were found attached to the external surface of the basal lamina, projecting into the hemocoel. Ectopic development of oocysts in the midgut epithelium between cells was observed much less frequently than development on the basal side of the midgut. The oocyst wall of dense granules, believed to be of parasite origin, was distinguishable from the basal lamina of the host's midgut epithelium. As in other Leucocytozoidae, the cytoplasm of the oocyst differentiated into a single sporoblastoid from which 30–50 sporozoites were formed. Beginning on the third day post infection, elongation of segregated dense sporoblastoid material associated with pellicle thickening led to the formation of the finger-like sporozoite buds which projected into the oocyst cavity. Sporozoites within mature oocysts and salivary glands were structurally similar to sporozoites as described for other haemosporidians.