福建及中国其他沿岸海域中国鲎资源分布现状调查

采用调访渔民、浅海底拖网、野外观测潮间带亲鲎产卵和幼鲎孵育情况等调查方法,重点调查了福建沿岸海域中国鲎(Tachypleus tridentatus)资源分布和数量,并走访调查了浙江省、广东省、海南省、广西壮族自治区和台湾地区金门岛等地代表性的中国鲎栖息地,对中国沿岸海域中国鲎资源现状作了较全面的调查。结果表明,大部分海滩已多年未见中国鲎上岸产卵,中国鲎在上岸之前已被渔民通过底拖网捕获,能见到幼鲎的海滩寥寥无几,中国鲎已经从大部分海域消失,中国鲎资源量濒临枯竭。目前尚存少量中国鲎的海域主要是东山湾和北部湾海域。金门岛海域则因为人工放流和设立了中国鲎保护区,中国鲎资源得到较好的恢复。中国鲎资源衰竭的主要原因是人类的过度捕捞、栖息地的破坏和海域环境污染等。中国鲎资源的保护迫在眉睫。建议将中国鲎列入国家级重点保护动物名录,增设中国鲎自然保护区,加强渔业管理以及人工增殖放流等措施。     

鲎肽素(tachyplesins)、tachystatins和polyphemusins

本文介绍分离自中国鲎、美洲鲎的三个肽类物质,即鲎肽素(tachyplesin)、tachystatin、polyphemusin及其氨基酸序列、生物活性、应用前景。     

基于MAXENT模型评估北部湾潮间带中国鲎和圆尾鲎稚鲎的潜在地理分布及种群保育对策

鲎具有极高的经济价值和科研意义。近年来由于过度捕捞和栖息地受损等原因,亚洲鲎种群数量正急剧下降。鲎漫长的生命周期使得鲎资源的保护和增殖迫在眉睫。生态位模型已经广泛应用于物种的潜在地理分布预测。基于实地调研数据和公开发表的北部湾中国海域中国鲎和圆尾鲎地理分布数据,运用MAXENT模型得到中国鲎和圆尾鲎在广西北部湾(中国部分)的栖息地适宜度指数(Habitat suitability index, HSI),确定了这两种稚鲎在北部湾中国海域潜在适生区。模型分析结果表明,潮间带坡度和地形指数是影响中国鲎分布的主要环境因子,而潮间带底质的有机物含量和植被指数是影响圆尾鲎分布的主要环境因子,根据研究结果建议在两种稚鲎适生区建立保护区,进行人工放流稚鲎,加强对海草和红树林的生态建设,进而促进鲎资源种群恢复和发展。     

鲎试验及其应用

<正> 本文就鲎试验及其应用的基本知识和进展,粗略介绍以下几个问题:(一)鲎及鲎试剂;(二)鲎试验的机理和方法;(三)关于鲎试验的评价及应用。     

中国鲎和圆尾鲎血淋巴细胞分类和特征的比较研究

为了更好地了解中国鲎(Tachpleus tridentatus)和圆尾鲎(Carcinoscorpius rotundicauda)血淋巴细胞的种类组成和特征差异,综合运用光学显微镜、扫描电镜和粒度仪,较为系统地对两种鲎的血淋巴细胞进行了分类和特征研究,从而为两种鲎的血淋巴细胞和分子生物学研究提供基础资料。根据血淋巴细胞大小、核质比、细胞着色特点、细胞中颗粒存在与否、颗粒的密集程度等,中国鲎和圆尾鲎的血淋巴细胞均可分为大颗粒细胞、小颗粒细胞和透明细胞三种主要类型,且两种鲎的血淋巴细胞均以颗粒细胞为主,透明细胞在血淋巴细胞中所占比例最小,但具有高核质比。两种鲎的同类血淋巴细胞在染色和形态上无显著性差异,但在同一种鲎中,血淋巴细胞密度存在显著的雌雄差异。       

鲎试剂在抗淋巴细胞免疫球蛋白热原检测中的应用

报道了鲎试剂在抗淋巴细胞免疫球蛋白内毒素检查中的应用。在用鲎试剂检测细菌内毒素时 ,按中国药典的要求对鲎试剂灵敏度进行标定 ,并作干扰实验 (增强或抑制实验 ) ,然后用鲎试法与家兔法同时测定 15批抗淋巴细胞免疫球蛋白。结果发现三批鲎试剂的灵敏度标示值均正确 ,五批抗淋巴细胞免疫球蛋白半成品在最大有效稀释度 (MVD)时对试验无干扰 ,这两种方法测定结果的符合率达到 91.7%。从而表明鲎试法很有可能代替家兔法     

全球中华鲎资源保护现状及对策建议

鲎是古老的海洋节肢动物。中华鲎(Tachypleus tridentatus)是世界现存4种鲎中体型最大的一种, 是河口生态系统的标志物种, 同时其血液被用于生产医用检验试剂――鲎试剂。中华鲎的自然地理分布范围相当狭窄, 仅局限于日本濑户内海向南延伸至印度尼西亚爪哇岛北岸以北的太平洋西岸海域, 其中在中国东岸和日本南部海域的历史产量较高。自20世纪50年代以来中华鲎种群数量出现了显著减少, 2019年中华鲎在IUCN红色名录中的濒危等级正式更新为濒危(EN), 明确了中华鲎资源呈现全球性衰退的状态, 究其原因可归纳为鲎生境破坏和过度捕捞两个方面。在开展鲎资源保护的实践工作中, 作者深刻反思当前鲎资源保护在海洋保护区划定、增殖放流及科普和野生动物保护法宣传中存在的问题并提出相应建议, 包括加快完善种群基线数据, 制定标准化种群和生境基线监测指南, 构建科学放流体系等, 以期推进全球范围内的中华鲎资源保护与科学管理。     

中国鲎变形细胞的超微结构观察

鲎属于节肢动物门螯肢动物亚门肢口纲剑尾目,现存三属五种,分布于北美、中美和东南亚沿海一带。变形细胞是鲎血液中最主要的细胞成分。 被称为活化石的鲎是一种古老的动物,在近四亿年的漫长过程中其形态几乎没有变化。自1956年Bang等给美洲鲎注射格兰阴性杆菌引起血管内凝血以来,鲎变形细胞溶解物用于微量内毒素的检测,受到普遍重视。近年来国内从中国鲎(Tachyplelus tridentatus     

鲎素抗菌肽作用机理研究

鲎素抗菌肽是中国鲎天然免疫的重要组成部分,具有广谱抗菌活性,对细菌、真菌、肿瘤细胞及一些病毒具有强大的抑制作用,且这种作用具有较好的选择性。本文主要介绍了鲎素的种类和生物活性、鲎素的结构与功能关系、鲎素作用机理的最新研究进展,并对鲎素临床应用面临的问题及对策做了简要综述。     

抗菌肽鲎素的研究进展

已报道的抗菌肽鲎素(tachyplesins,TPs)共分5种,均由鲎血细胞分离产生,是鲎先天免疫系统的重要功能物质,为鲎机体抵抗病原物入侵提供天然防御屏障。近30年的研究表明,TPs抗菌肽显示出广谱生物活性、较低正常细胞毒性、不易产生微生物抗性等特点,作为一种全新型的肽类药物备受关注。文中主要从TPs的分子结构,对微生物、藻类细胞、肿瘤细胞及一些病毒的抑杀活性,作用机制,以及工程表达情况等方面进行介绍,综述TPs的应用前景,并对当前研究瓶颈进行评述。     

Investigations on the Specificity of the Limulus Test for the Detection of Endotoxin

Lysates obtained from amoebocytes of Limulus polyphemus, the horseshoe crab, showed gel formation after the addition of bacterial endotoxin. In contrast to living gram-negative bacteria, viable gram-positive microorganisms did not cause gelation of lysate. Nevertheless, peptidoglycan isolated from the cell walls of various gram-positive organisms did induce the reaction. However, the activity of peptidoglycan was 1,000 to 400,000 times less than that of Escherichia coli lipopolysaccharide. After exposure to lysozyme, peptidoglycan no longer gelled amoebocyte lysate, therefore apparently excluding endotoxin contamination. Gelation of amoebocyte lysate by endotoxin or peptidoglycan was inhibited by different concentrations of sodium polystyrolsulfonate. Whereas these studies confirm the specificity of the Limulus test for bacterial endotoxins, they also indicate that other substances of bacterial origin should be investigated for their ability to gel amoebocyte lysate.     

Differentiation of immune cells challenged by bacteria in the common European starfish,Asterias rubens (Echinodermata)

Amoebocytes are the main effector cells of the echinoderm immune system. In starfishes, a taxon in which bacterial diseases have been rarely reported, amoebocytes are considered to be the only circulating and immune cell type. The present paper addresses the question of amoebocyte differentiation in the starfish Asterias rubens when challenged by bacteria. Starfishes were injected with FITC-coupled bacteria (Micrococcus luteus). Amoebocytes were collected at regular time intervals for 24 h. The cytometric characteristics and the phagocytic activity were studied by flow cytometry. Three amoebocyte groups of different size were identified. The cell concentrations of the two largest and more numerous of these groups (G2 and G3) were modulated by immune stimulation while the group of smallest, less numerous, cells (G1) was unaffected. All of these cell groups were phagocytic but their kinetics of cell activation and bacteria ingestion differed. G1 cells showed the lowest phagocytic activity while G3 cells had the highest and fastest phagocytic activity. Starfish amoebocytes appear to be segregated in three groups, two of them (G2 and G3) being immunomodulated and one of them presenting a very fast reaction to bacteria. It is suggested that the high efficiency of the immune system in starfishes is related to this fast reaction.     

Immunocytochemical demonstration of a humoral defence factor in blood cells (amoebocytes) of the pond snail,Lymnaea stagnalis

Summary Monolayers of blood cells (amoebocytes) and sections of connective tissue and of amoebocyte pellets of the freshwater pulmonate gastropod Lymnaea stagnalis were stained immunocytochemically with antisera to snail agglutinin/opsonin. The presence of this substance was demonstrated light microscopically both in the cytoplasm and on the cell membrane of amoebocytes. This suggests that amoebocytes synthesize agglutinin/opsonin, and bear it at their surface as receptors for foreign materials.     

Qualitative and quantitative study of wound healing processes in the coelenterate, Plexaurella fusifera: spatial, temporal, and environmental (light attenuation) influences

Following injury, the Caribbean soft coral, Plexaurella fusifera, forms an epithelial front containing amoebocytes and zooxanthellae, a photosynthetic endosymbiont. Amoebocytes may be responsible for extruding the connective mesogleal fibers necessary for regeneration of tissue and zooxanthellae may provide the energy for repair. This study examined the effects of time, space, and environment (light attenuation) on wound healing in this coral species and quantitatively confirmed the increase of amoebocyte concentrations in the injured area. A wound was made on coral branchlets by removing approximately 4.5 mm of coenenchyme. At assigned times after injury, samples were collected for gross morphological and histological evaluation, in which amoebocytes and zooxanthellae concentrations were quantified within 0.009 mm3 of tissue. Overall amoebocyte numbers within uninjured and wounded tissue were similar. However, when numbers of amoebocytes per area of injured tissue were calculated and compared to those of uninjured tissue, 82.4% more amoebocytes occurred at distances 0-0.5 mm from the wound edge, while areas of tissue >2 mm from the wound edge were occupied by fewer amoebocytes. Overall increases in concentrations of zooxanthellae also occurred within wounded coral, but no apparent temporal, spatial, or light-related pattern was detected. Therefore, this study supports the conjecture that amoebocyte accumulation at a wound site is an effect of cells migrating from uninjured tissue adjacent to the wounded edge. In addition, this movement occurs regardless of light attenuation. Light, which in this study was confined to ranges between 70 and 545 microE s-1 m-2, did not significantly affect the wound healing process in regard to either closure or cellular concentrations.     

Ultrastructure of Holothuria polii encapsulating body

The coelomic cavity of freshly collected Holothuria polii specimens contains a variable number of brown pigmented and unpigmented encapsulating structures. They are composed of nodules with entrapped parasites and an internodular mass which comprises a number of nodules. Both brown and unpigmented bodies occur in different size classes depending upon the number of nodules accumulated in a complete body. The unpigmented bodies probably represent an early unmelanized stage of the brown ones.
The nodule was ultrastructurally constituted by foreign bodies surrounded by a fibrous, electron-dense, non-cellular layer, probably melanin, followed by a layer of elongated and extremely flattened amoebocytes. Nodules were assembled in an internodular mass formed by amoebocytes type I, II and III spherule cells. As for the amoebocytes constituting the nodule, those present in the spaces between nodules did not develop junctional complexes. It is conceivable that the intricate network established among the cell processes could represent the mechanical force maintaining the whole structure.
As suggested by our results, two functional amoebocyte populations seem to be responsible for the organization of the scavenger body: 1. encapsulating amoebocytes, characteristically non phagocytosing, elongated cells; and 2, phagocytosing amoebocytes. The former organize the nodules, the latter constitute the internodular mass of several nodules.
Most probably, the double scavenger activity justifies the considerable complexity of the H. polii encapsulating structure compared to other invertebrates.
Spherule cells participate only in constituting the internodular mass.     

Schistosoma mansoni: immunofluorescent detection of its antigen reacting with Biomphalaria glabrata amoebocytes

The reaction of amoebocytes in the hemolymph of the infected intermediate host, Biomphalaria glabrata, to Schistosoma mansoni antigens has been investigated using the indirect immunofluorescence antibody test. Monolayers of amoebocytes, prepared from hemolymph of infected and normal snails, were first fixed and then reacted with antisera obtained from mice infected for 7 to 9 weeks. Nonspecific and cross-reactions between the antisera and monolayers of amoebocytes were eliminated by absorbing the antisera with tissues from uninfected snails. The liberation of detectable schistosomal antigens in the hemolymph in soluble and particulate forms coincided with completion of the infection cycle 3 to 4 weeks after exposure to miracidia. The schistosomal antigens were demonstrable in the cytoplasm of amoebocytes and in the center of amoebocyte aggregates.     

Cellular responses in sea fan corals: granular amoebocytes react to pathogen and climate stressors

Background

Climate warming is causing environmental change making both marine and terrestrial organisms, and even humans, more susceptible to emerging diseases. Coral reefs are among the most impacted ecosystems by climate stress, and immunity of corals, the most ancient of metazoans, is poorly known. Although coral mortality due to infectious diseases and temperature-related stress is on the rise, the immune effector mechanisms that contribute to the resistance of corals to such events remain elusive. In the Caribbean sea fan corals (Anthozoa, Alcyonacea: Gorgoniidae), the cell-based immune defenses are granular acidophilic amoebocytes, which are known to be involved in wound repair and histocompatibility.

Methodology/Principal Findings

We demonstrate for the first time in corals that these cells are involved in the organismal response to pathogenic and temperature stress. In sea fans with both naturally occurring infections and experimental inoculations with the fungal pathogen Aspergillus sydowii, an inflammatory response, characterized by a massive increase of amoebocytes, was evident near infections. Melanosomes were detected in amoebocytes adjacent to protective melanin bands in infected sea fans; neither was present in uninfected fans. In naturally infected sea fans a concurrent increase in prophenoloxidase activity was detected in infected tissues with dense amoebocytes. Sea fans sampled in the field during the 2005 Caribbean Bleaching Event (a once-in-hundred-year climate event) responded to heat stress with a systemic increase in amoebocytes and amoebocyte densities were also increased by elevated temperature stress in lab experiments.

Conclusions/Significance

The observed amoebocyte responses indicate that sea fan corals use cellular defenses to combat fungal infection and temperature stress. The ability to mount an inflammatory response may be a contributing factor that allowed the survival of even infected sea fan corals during a stressful climate event.     

Biomphalaria glabrata amoebocytes: Assay of factors influencing in vitro phagocytosis

A simple and short-term in vitro assay system has been adapted for quantitative evaluation of the rate of phagocytosis of amoebocytes obtained from hemolymph of the pulmonate gastropod Biomphalaria glabrata. The variation in the rate of phagocytosis exhibited by amoebocyte monolayers prepared from individual snails is reduced by utilizing amoebocytes from pooled hemolymph from snails of similar sizes. The study demonstrates that the rate of phagocytosis depends on incubation time, temperature, and pH. Substantial inhibition of phagocytosis is exerted by 2-deoxy-d-glucose and 2-iodoacetamide but not by potassium cyanide, indicating that energy consumed during phagocytosis is generated by the glycolysis pathway.     

中国鲎变形细胞超微结构的进一步观察

中国鲎的变形细胞可在血液中发育成熟,根据其超微结构,可分为原始变形细胞、幼变形细胞和变形细胞三个阶段。 变形细胞受到外环境或内毒素的刺激后,第一型颗粒出现一系列变化,最终把内容物释放到细胞外。与此同时,细胞表面形成若干胞质性突起,边缘带分散断裂,质膜下和胞质性突起中出现微丝。第二型颗粒无变化,提示它们和血液凝胶化过程无直接关系。