强壮粗体虫的线粒体基因组及棘头虫的系统发育研究简

通过长距离PCR方法,克隆了鳜(Siniperca chuatsi Basilewsky)肠道内寄生虫——强壮粗体虫(Hebesoma violentum Van Cleave)线粒体基因组全长序列,共13393 bp(GenBank登录号:KC415004),有36个基因,其中蛋白编码基因12个,核糖体基因2个,tRNA22个。所有基因均由线粒体基因组同一条链按同一个方向转录。利用该线粒体基因组和已经报道的一些轮虫纲种类的线粒体基因组序列,构建了棘头虫和轮虫的系统发育树。系统发育研究表明:包括强壮粗体虫、隐藏新棘虫Pallisentis celatus(Van Cleave)和Paratenuisentis ambiguous(Van Cleave)在内的始新棘头虫纲(Eoacanthocephala)与古棘头虫纲(Palaeacanthocephala)亲缘关系较近,聚为一枝后再与原棘头虫纲(Archiacanthocephala)聚在一起;棘头虫与双巢类轮虫(Bdelloid)亲缘关系最近,聚为一枝,然后再与单巢类轮虫(Monogonont)聚在一起,表明棘头虫和轮虫具有较近的亲缘关系。     

大熊猫蠕形螨病组织病变的初步观察

大熊猫蠕形螨病的临床症状和危害已有记述〔1,2〕。本文通过一例病猫皮损活组织块,经石蜡包埋做成连续切片,HE染色,厚度为8μm,镜下见到表皮角质层增厚,棘层稍增厚,基层正常。毛囊漏斗部与皮脂腺导管都有不同程度的扩大,其内毛根脱落较多。同时,被许多蠕形螨碎片及完整的蠕形螨充塞。部分毛囊的上皮细胞有坏死脱落、核溶解及核碎裂现象;有的含虫毛囊周围的间质内有大量嗜中性粒细胞及嗜酸性粒细胞浸润;个别毛囊腔内充满大量嗜中性粒细胞及脓球;少数汗腺管内及其它结缔组织内也有炎性细胞浸润。组织内大熊猫蠕形螨主要寄生于毛囊上段,即漏斗…     

中国对虾中肠的超微结构与细胞化学研究

(1)在透射电镜下观察中国对虾 (PenaeuschinensisOsbeck)中肠肠壁的超微结构。结果显示 :中国对虾中肠肠壁由内向外依次为上皮细胞层、结缔组织层、肌层和结缔组织层四层结构。中肠上皮有明、暗两种细胞 ,以暗细胞占多数 ,两种上皮细胞都具有从肠腔吸收营养物质和向结缔组织中转运营养物质的功能。 (2 )运用酶细胞化学技术 ,显示中肠组织细胞中的酸性磷酸酶 (ACP)和酚氧化酶 (PO)活性 ,电镜观察显示 :ACP阳性反应出现在明细胞顶部的圆形小泡和明、暗两种细胞的溶酶体中 ,暗细胞顶部聚集着大量ACP阴性反应的酶原颗粒 ,酶原颗粒主要由暗细胞合成并分泌。PO颗粒出现在患病虾肠壁结缔组织内解体的颗粒细胞上和游离于结缔组织中。结缔组织除具有维持上皮细胞的形态、贮存营养物质的功能 ,还在对虾免疫反应中发挥作用     

方斑东风螺单孢子虫病的研究

文章对方斑东风螺(Babylonia areolata Link, 1807)单孢子虫病进行了首次报道, 对其病理学特征、流行季节和诊断方法进行了研究。结果认为方斑东风螺被单孢子虫侵袭后, 吻管、足肌、肠、消化腺、鳃、胃和肝等器官引起一系列病变。病理组织切片观察到肠上皮黏膜组织细胞布满该虫营养体和各发育期的多核质体, 造成肠结缔组织呈现炎症反应。寄生部位病灶肿胀、混浊、坏死、崩解。       

圆形碘泡虫和关桥碘泡虫孢子的光镜及扫描电镜观察

对寄生于鲫的圆形碘泡虫(Myxobolus rotundus Nemeczek,1911)及异育银鲫的关桥碘泡虫(M．guanqiaoensis Wu ＆Wang,1997)的成熟孢子进行光镜及扫描电镜观察。圆形碘泡虫主要寄生于鲫体表、头部、鳃、吻部及鳍条,形成许多大小不一的乳白色胞囊。关桥碘泡虫主要寄生于异育银鲫的肝脏,肝脏基本上被虫体所充满,肝脏组织被破坏。扫描电镜观察表明圆形碘泡虫成熟孢子表面光滑,缝脊直而明显,宽0．2μm,缝脊上下两侧对称。关桥碘泡虫成熟孢子表面皱褶,其程度不等,有的虫体一侧向内凹陷形成盆状,缝脊呈“S”形,宽0．5μm,缝脊上下两侧不对称,其中间有一不很明显凹陷的沟。     

小鼠中性粒细胞在抗体及补体参与下对日本血吸虫童虫杀伤作用的体…

用光镜及电镜观察小鼠中性粒细胞及中性粒细胞依赖抗体及补体对体外培养的日本血吸虫童虫的作用。结果表明,单纯中性粒细胞很少粘附到童虫表面,仅个别发疏松地粘附在童虫表面,被粘附的童虫结构正常,提示：单纯中性粒细胞对童虫无明显作用,在抗体及补体协同下,中性粒细胞成群且紧密地粘附在童虫体表,在细胞集聚的周围,虫体体被出现隧道样及火山口样变化,紧贴童虫的中性粒细胞伸出伪足,虫体体棘紊乱,皮层变平,体被剥脱,虫     

南海石斑鱼苗种肠道微孢子虫病病原的鉴定

研究通过组织病理分析、超微结构观察以及分子特征分析对石斑鱼(Epinephelus spp.)苗种肠道微孢子虫病病原进行了鉴定。其为一肠孢虫属新种, 命名为石斑鱼肠孢虫(Enterospora epinepheli sp. n.), 专性寄生于细胞核内, 发育过程与肠孢虫属模式种黄道蟹肠孢虫(Enterospora canceri)一致。早期单核裂殖体通过一层简单的电子薄膜与宿主细胞核质隔离。随后, 单核裂殖体发育形成多核裂殖原质团。此时, 细胞核出现明显肥大, 有的甚至被裂殖子胀破。裂殖原质团进一步发育形成多核产孢体, 并开始出现许多高电子密度的囊泡状结构。这些与极丝及锚状盘有关的囊泡状结构聚集在藕核周围, 并组装形成微孢子虫特征性结构(挤出装置)前体。随后, 产孢体原生质团通过连续分裂形成一个个孢子母细胞。孢子母细胞与细胞核直接接触, 并直接发育形成成熟孢子。成熟孢子椭圆形, 孢子长(1.56±0.31) μm (1.07—1.96 μm), 宽(1.08±0.98) μm (0.93—1.28 μm)。 孢壁分为3层, 外壁电子密度高, 厚(15.51±0.95) nm (9.87—26.18 nm), 内壁为电子透明层, 较外层更厚(81.13±2.71) nm (57.16—110.81 nm), 最里面为孢质膜。极丝为同型极丝, 共5—6圈, 分2排排列。组织病理学分析发现该微孢子虫寄生于肠道上皮杯状细胞核内, 肠壁脱落的内容物中也发现大量的微孢子虫。序列比对发现该种与之前报道的石斑鱼肠道微孢子虫待定种(Microsporidium sp.)序列基本一致, 与其他相似性较高的种类的遗传距离在0.162—0.225。系统发育关系分析显示肠胞虫科的种类明显分为两支, 石斑鱼肠孢虫和肠孢虫属其他种类及毕氏肠胞虫聚为一个独立分支, 但不与该分枝中任何种类形成姊妹支。     

多疣壁虎肠道内分泌细胞的分布及形态学观察

用光镜和电镜对多疣壁虎肠道嗜银细胞的分布及形态做了初步的观察,结果显示,十二指肠嗜银细胞密度最高,大肠次之,空、回肠较低;作者认为嗜银细胞的分布与动物生活环境是相关的,嗜银细胞形态多样,位于肠上皮细胞之间和固有层结缔组织中,电镜下,嗜银细胞内充满足电子致密颗粒,十二指肠和大肠嗜银细胞颗粒大小和密度不同。根据内分泌特点的不同,可分为开放型和闭合型两类。本文还对多疣壁虎肠道嗜银细胞的分布及形态特点做了探讨。     

小鼠中性粒细胞在抗体及补体参与下对日本血吸虫童虫杀伤作用的体外实验

用光镜及电镜观察小鼠中性粒细胞及中性粒细胞依赖抗体及补体对体外培养的日本血吸虫童虫 的作用。结果表明：单纯中性粒细胞很少粘附到童虫表面,仅个别十分疏松地粘附在童虫表面,被粘附 的童虫结构正常。提示：单纯中性粒细胞对童虫无明显作用,在抗体及补体协同下,中性粒细胞成群且 紧密地粘附在童虫体表,在细胞集聚的周围,虫体体被出现隧道样及火山口样变化,紧贴童虫的中性 粒细胞伸出伪足,虫体体棘紊乱,皮层变平,体被剥脱,虫体变形,说明中性粒细胞在抗体及补体协同 下,对童虫有杀伤作用、文中对杀伤机制进行了扼要的讨论。     

鲤肠道寄生岳阳碘泡虫新种的形态特征及系统发育分析

在洞庭湖岳阳地区开展鱼类寄生虫调查中,发现一种寄生于鲤Cyprinus carpio L.肠道的黏孢子虫。该黏孢子虫的孢囊呈白色,椭圆形,大小为(1.0±0.2) mm (0.8—1.2 mm)。成熟孢子具有壳瓣,壳面观近似圆形,后端有4—6个“V”形褶皱;缝面观呈纺锤形,缝脊直而粗;孢质均匀,含有一个嗜碘泡;孢子长(9.8±0.6) μm (9.6—10.0 μm),孢子宽(8.2±0.3) μm (8.0—8.5μm),孢子厚(7.3±0.1) μm (7.0—7.5 μm);2个极囊梨形,位于孢子顶端,大小相等,呈“八”字形;极囊长(4.4±0.4) μm (3.8—5.1 μm),宽(2.7±0.2) μm (2.2—3.2 μm),极丝4—5圈。该黏孢子虫与肠膜碘泡虫、丑陋圆形碘泡形态特征非常相似,但其极囊/孢子小于1/2;与文献已报道的鲤肠道寄生北京碘泡虫和鲤肠碘泡虫相比较,其在孢子形态、孢子和极囊大小方面分别存在明显差异。基于该黏孢子虫18S rDNA基因序列(GenBank登录号KY203795)比对分析,该黏孢子虫与山东碘泡虫相似率最高,仅为96%。系统发育分析发现,该黏孢子虫与山东碘泡虫、倪李碘泡虫、住心碘泡虫、Myxobolus encephalicus、Sphaerospora molnari、多涅茨尾孢虫和Henneguya zikaweiensis聚为独立分支,和其他已报道的黏孢子虫亲缘关系较远。综合形态学和18S rDNA基因序列数据,文章报道的鲤肠道寄生黏孢子虫为碘泡虫属一新物种,将其命名为岳阳碘泡虫。     

Intestinal pathology and site specificity of the acanthocephalan Neoechinorhynchus carpiodi Dechtiar, 1968, in quillback, Carpiodes cyprinus (Lesueur)

Intestinal pathology and site specificity of Neoechinorhynchus carpiodi Dechtiar, 1968, in quillback, Carpiodes cyprinus (Lesueur), from Dauphin Lake, Manitoba, was investigated. Neoechinorhynchus carpiodi elicits the formation of a nodule at the point of proboscis attachment. Over 98% of the worms were attached in sections 6-8 of the intestine. Fish had clusters of 1-8 nodules, cluster size increased with worm burden, and the volume of nodules increased with worm numbers. Nodule number, size, and pathology was related to intensity of infection and to depth of proboscis penetration. Three types of pathology were noted. Type I nodules had few worms and the proboscides were attached in the lamina propria; Type II nodules harbored more worms and the proboscides penetrated to, but not through, the stratum compactum; and Type III nodules had the greatest numbers of N. carpiodi and proboscides penetrated through the stratum compactum. Hyperplasia, vascularization, and collagen deposition were extensive, especially in Type III nodules. An epithelium-lined channel was present in most nodules. Type III nodules were most anterior (75% of the time), harbored the greatest proportion of immature worms regardless of cluster size, and had more gravid females. The largest nodules (Type III pathology) were well vascularized, long-lasting, and appeared to be the preferred microhabitat for the parasite.     

Trichinella spiralis: peroxidase activity in isolated cells from the rat intestine

An understanding of physiologic events underlying resistance to parasitic worms depends on a knowledge of metabolic interactions between parasites and specific cells at the host-parasite interface. In the case of invasive intestinal parasites this interaction involves contact with epithelial cells and cells of the lamina propria. This investigation deals with the collection of epithelial cells and lamina propria cells from the small intestine of control rats and rats infected with the nematode, Trichinella spiralis, and measurement of peroxidase activity in these cells. Lamina propria cells were isolated by collagenase digestion of everted gut segments previously denuded of epithelium by treatment with hyaluronidase. Mean peroxidase activity in homogenates of lamina propria cells was equivalent to 40 nmoles H₂O₂ decomposed/min/mg of cell protein from control rats compared to 413 nmoles from infected animals. Epithelial cell peroxidase activity in homogenates of epithelial cells from both control and infected rats was less than 2 nmoles H₂O₂ decomposed/min/mg cell protein. The degree of contamination of lamina propria cells with epithelial cells was determined by measuring disaccharidase activity in both cell populations. The specific activity of maltase, sucrase, and trehalase in lamina propria cells was between 10 and 17% of that in epithelial cells. This work is a requisite for a study in which the role of intestinal cell peroxidase in resistance to Trichinella will be evaluated.     

The ontogeny of gut-associated lymphoid tissue in Oreochromis-mossambicus

The ontogenesis of gut-associated lymphoid tissue (GALT) was investigated using histological procedures. Samples were taken at various time intervals to include larval, first-feeding and juvenile stages. Whole fish and tissue samples were embedded in either paraffin wax or metha-crylate resin and sections stained with Giemsa, periodic acid-Schiff (PAS), methyl green pyronin (MGP) and methylene blue. GALT was first observed as individual lymphocyte-like and macrophage-like cells in the lamina propria, and in later stages plasma cells and granulocytes were identified. Leucocytes began to infiltrate the epithelium just prior to first feeding and then rapidly once feeding had began. By day 50, GALT was equal to that observed in the adult gut.     

Ileal mucosal mast cell, eosinophil, and goblet cell populations during Hymenolepis diminuta infection of the rat.

The population dynamics in the enteric connective tissues of eosinophils, mucosal mast cells (MMC), and in the mucosal epithelium of goblet cells were examined morphometrically in fixed ileal tissue of outbred Sprague Dawley rats during the first 32 days of infection with the tapeworm Hymenolepis diminuta. MMC and eosinophils were present in the lamina propria and submucosa; however, only eosinophils were also present in the muscularis externa. Eosinophilic infiltrate was first observed in the lamina propria at 15 days postinfection (dpi) and the numbers of eosinophils remained elevated through 32 dpi. Initial mucosal mastocytosis was detected on 6 dpi and MMC numbers continued to rise over the study period without reaching a plateau. Goblet cell hyperplasia occurred only at 32 dpi. In contrast to some intestinal nematode infections where these same 3 cell types are associated with the host's expulsion responses, H. diminuta is not lost by a rapid host response in the outbred Sprague Dawley rat strain used in these experiments. We suggest that either the induction of hyperplasia of these host effector cells in ileum tissue during H. diminuta infection is not capable of triggering parasite rejection mechanisms, or the function of the induced hyperplasia is necessary for some as yet unassociated physiological or tissue architecture change in the host's intestine.     

Invasiveness of Helicobacter pylori into Human Gastric Mucosa

Background. Helicobacter pylori has generally been observed only in the gastric mucous layer or in the spaces between gastric mucus -s ecreting cells and not in the gastric epithelial cells or in the lamina propria. The purpose of this study is to determine whether H. pylori invades the gastric mucosa, using an immunoelectron microscopical examination of human gastric mucosa infected with H. pylori.
Materials and Methods. Five hundred gastric antral biopsy specimens were fixed in a periodate-lysin-paraformaldehyde solution, embedded in Lowicryl, sectioned, and examined with a light microscope. One hundred specimens moderately or severely infected with H. pylori were selected and were incubated with polyclonal rabbit anti– H. pylori antibody. The specimens were washed, incubated with 20 nm of colloidal gold–conjugated goat anti–rabbit IgG, stained with uranyl acetate and lead citrate, and observed with a transmission electron microscope.
Results. In one case, a bacterium was observed within the cytoplasm of a gastric mucus -s ecreting cell; in another case, a few bacteria were observed within the cytoplasm of a stromal cell in the lamina propria. The bacteria could be differentiated from degenerated intracellular organelles by gold particles attached to the bacteria.
Conclusion. H. pylori rarely invade the lamina propria and gastric cells.     

Immunohistochemical study on arachidonate 5-lipoxygenase in porcine leukocytes and other tissues

Arachidonate 5-lipoxygenase is an enzyme that catalyzes the oxygenation of arachidonic acid, producing 5-hydroperoxy acid. This enzymatic reaction initiates the biosynthesis of various bioactive leukotrienes. An antiserum was raised in a rabbit against the purified 5-lipoxygenase of porcine leukocytes, and various types of porcine leukocytes were immunostained by use of the antibody. As examined by light and electron microscopy, neutrophils and eosinophils were positively stained. The 5-lipoxygenase was localized in the cytoplasm but not in the plasma membrane and subcellular organelles of the positively stained cells. In contrast, lymphocytes were unstained. In porcine ileum, the majority of 5-lipoxygenase-positive cells were eosinophils and mast cells resident in the lamina propria mucosae, whereas parenchymal cells were not stained. In porcine lung, certain bronchiolar or bronchial epithelial cells were clearly immunostained, in addition to eosinophils and mast cells found in the interstitium.     

THE DISTRIBUTION AND DIFFERENTIATION OF LYMPH-BORNE IMMUNOBLASTS AFTER INTRAVENOUS INJECTION INTO SYNGENEIC RECIPIENTS

Thoracic duct lymph from inbred, hooded rats was collected 3–5 days after antigenic stimulation of the caudal lymph nodes. During this period the lymph contained 10–15% of large, basophilic lymphoid blast cells (immunoblasts). By incubating the lymph cells at 38.C with radioactive DNA precursors, either ³H-thymidine or ¹²⁵I-deoxyuridine, the immunoblasts became labelled but the small lymphocytes did not. The lymph cells were then washed and injected intravenously into syngeneic recipients which were killed after various intervals up to 24 hr so that the radioactivity of their organs could be assayed by scintillation counting and autoradiography.
The main finding was that in animals killed after 4 or more hours the small gut always contained most of the recoverable activity and autoradiographs showed that this was because the injected cells had infiltrated the lamina propria in large numbers. Earlier, many of the injected cells were retained temporarily in the lungs, liver and spleen but many of them soon left those organs and entered the lamina propria of the small gut.
An electron microscope study of autoradiographs showed that 24 hr after injection the cells which entered the lamina propria of the gut had differentiated into plasma cells so that they displayed abundant, lamellar endoplasmic reticulum.     

Distribution of Leu 7 (HNK-1) antigen in human digestive organs: an immunohistochemical study with monoclonal antibody

Summary Leu 7 immunoreactivity was demonstrated with the indirect peroxidase-labelled antibody method on frozen and paraffin-embedded tissue sections of human digestive organs. Anti-Leu 7 monoclonal antibody, which allegedly detects mononuclear cells with natural killer or killer activity, recognized lymphoid cells among intestinal epithelial cells and in the germinal centres of solitary lymphoid follicles of small and large intestine, and a few in gallbladder, liver and the lamina propria of the intestine. In addition, peripheral nerve fibres, endocrine cells in the gut and pancreas and carcinoid and islet cell tumours were also positively stained. At the ultrastructural level, Leu 7 antigen was localized on the plasma membrane of granulated lymphoid cells in the gut mucosa and on the secretory granules of intestinal endocrine cells. In normal pancreas, Leu 7 immunoreactivity was demonstrated in most cells containing pancreatic polypeptide and in many cells containing somatostatin or glicentin. Insulin-containing cells, however, lacked Leu 7 immunoreactant. These findings were obtained in both frozen sections and paraffin-embedded sections. The possible cross-reactivities of monoclonal antibodies are discussed as they raise an important caveat in immunohistochemical studies using these antibodies.     

Transepithelial pathogen uptake into the small intestinal lamina propria

The lamina propria that underlies and stabilizes the gut lining epithelium is densely populated with strategically located mononuclear phagocytes. Collectively, these lamina propria macrophages and dendritic cells (DC) are believed to be crucial for tissue homeostasis as well as the innate and adaptive host defense. Lamina propria DC were recently shown to gain direct access to the intestinal lumen by virtue of epithelium-penetrating dendrites. However, the role of these structures in pathogen uptake remains under debate. In this study, we report that entry of a noninvasive model pathogen (Aspergillus fumigatus conidia) into the murine small intestinal lamina propria persists in the absence of either transepithelial dendrites or lamina propria DC and macrophages. Our results suggest the existence of multiple pathogen entry pathways and point at the importance of villus M cells in the uptake of gut lumen Ags. Interestingly, transepithelial dendrites seem altogether absent from the small intestine of BALB/c mice suggesting that the function of lamina propria DC extensions resides in their potential selectivity for luminal Ags, rather than in general uptake or gut homeostasis.     

A monoclonal antibody specific for rat intestinal lymphocytes

A monoclonal antibody, RGL-1, was produced by fusion of NSI myeloma cells with spleen cells of a mouse immunized with isolated rat intraepithelial lymphocytes (IEL). SDS-PAGE analysis revealed that RGL-1 precipitated two major noncovalently bound chains of about m.w. 100,000 and 125,000, and a minor component of m.w. 200,000. Examination of both tissue sections and isolated cells indicated that RGL-1 stained the majority of the lamina propria lymphocytes and IEL but only very few cells (less than 2%) in the lymphoid organs and small numbers of lymphocytes in other mucosae. In the small intestine, RGL-1 stained lymphocytes with the helper (W3/25) as well as the cytotoxic/suppressor (OX8) phenotype. The antibody reacted with 95% of the granular IEL but with less than 0.1% of the blood large granular lymphocytes. Although mature IgA plasma cells in the lamina propria were RGL-1-, some large IgA-containing cells were weakly positive. In the gut-associated lymphoid tissues (GALT), studies combining immunofluorescence and autoradiography indicated that 56 and 73% of rapidly dividing cells of mesenteric lymph nodes and of thoracic duct lymph (TDL) stained with RGL-1, respectively. In addition, 90 to 100% of the IgA-containing blasts of MLN and 75% of those of TDL were labeled by RGL-1. In contrast, rapidly dividing cells of spleen and of peripheral lymph nodes did not stain with RGL-1. Because RGL-1 can be demonstrated on both intestinal lymphocytes and their immediate precursors in the GALT, its expression may be related to the homing of lymphocytes into the gut mucosa.