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。     

鲎的分布及生活习性

鲎是一种古老的节肢动物，现存的鲎分为二亚科三属四种。本文对鲎在世界上的分布及其生活习性做了概述。     

中国鲎幼体阶段(黄皮鲎)的形态特点

经研究发现,中国鲎幼体阶段（黄皮鲎）生殖盖板中叶不长于侧叶,尾呈三角形,上有一列小刺,腹甲后端尾上有三个棘突,尾两个侧边无小刺,腹甲背面无小刺分布,但其头胸甲和腹甲背面棘突较长,明显呈幼体状,中国鲎的胚后幼体发育从小到大,身体背部颜色处于黑色→土黄色→褐色的一种渐变过程,当中国鲎幼体头胸甲宽度处于10-25cm时,也就是胚后发育的第12至第16次蜕皮期间时,其体色呈土黄色,在粤西、海南、北部湾一带俗称“黄皮鲎”。所以,黄皮鲎为中国鲎一定阶段未成熟的幼体,而不是一个新种或亚种。标本采自广东省湛江市,保存于湛江海洋大学水生生物博物馆。     

鲎人工授清试验的初步观察

本文报告东方鲎的人工授精实验,共进行四次,均获得成功,受精率述9 5％以上,经49—59天孵育出幼鲎。文中较详细地介绍了人工授精的方法及注意事项,并对鲎胚的发育过程做简单介绍。     

四种重金属对中国鲎(Tachypleus tridentatus)胚胎发育的影响

分析了不同离子浓度的Zn、Pb、Cu、Cd对中国鲎胚胎发育的影响.结果表明①4种金属离子浓度≤渔业水域水质标准浓度4倍时,对中国鲎胚胎发育无明显毒性效应;≥渔业水域水质标准浓度8倍时,Cd离子和Cu离子对中国鲎胚胎发育无明显影响,而Zn离子组Pb离子组胚体发育随离子浓度提高而卵径变小,发育速度下降;②浓度为1.6mg/L时,Zn和Pb离子组胚胎致畸率分别为44%和58%,Pb离子组胚体孵化率仅2.67%;③4种重金属对中国鲎胚胎发育的毒性效应表现为Pb＞Zn＞Cu=Cd;④中国鲎胚胎对重金属有蓄积作用,蓄积效应随金属种类不同而异.     

中国鲎血细胞原代培养及部分生物学现象观察

中国鲎（Tachypleustridentatus）,隶属于节肢动物门（Arthropoda）、有螯亚门（Chelicerata）、肢口纲（Merostomata）、剑尾目（Xiphosura）、鲎科（Limulidae）,分布于中国、日本和东南亚其他国家。鲎科动物（Horseshoecrab）现有三属四种,自从早古生代奥陶纪出现,至今4亿多年,其形态结构以及生活习性无重大变化。在胚胎发育后期有三叶虫幼体的形态特征,被誉为活化石。在天然免疫、视觉、胚胎发育等许多方面,鲎成为研究者的模式动物。     

鲎源抗菌肽的研究及其潜在应用价值

鲎血细胞来源的抗菌肽,在鲎天然免疫中起至关重要的作用,它对外源病原菌具有抗性作用,降低了外源病原菌对鲎活体的致病性,增强了鲎的天然免疫能力。鲎源抗菌肽有一些不同于其他来源的抗菌肽的优势,对鲎源抗菌肽的研究有很重要的实用意义。概述了鲎源抗菌肽的性质、分子结构、基因序列及其制备,并对它们的潜在应用价值进行了论述。     

中国鲎凝集素的分离纯化及性质研究

 经N-乙酰氨基葡萄糖交联琼脂糖亲和层析及以交联琼脂糖介质的高效液相分子筛层析,从中国鲎细胞溶解物中分离纯化了一种凝集素,其活性比原料鲎试剂提高128倍。鲎凝集素SDS电泳时表现出分子量为69000,和72000的二个亚基。N-乙酰氨基葡萄糖、D-半乳糖,D-甘露糖及岩藻糖等对鲎凝集素凝集鸡红细胞的活性有显著抑制作用,加热60℃,10分钟可使凝集素活性基本丧失。CaCl_2为凝集素活性所必需。鲎凝集素与肺炎球菌C多糖有沉淀反应。     

Position of horseshoe crabs in estuarine food webs: N and C stable isotopic study of foraging ranges and diet composition

To discern the position of horseshoe crabs as a potentially important predator in estuarine food webs, we determined where they foraged and what they ate. We used N and C stable isotopes to link adult horseshoe crabs to their oraging locations and potential food sources in Pleasant Bay, Cape Cod. The δ¹⁵N in tissues of horseshoe crabs and their potential foods suggest crabs were loyal to local foraging sites and did not forage substantially in subestuaries receiving >110 kg N ha⁻¹ year⁻¹. Among locations where crabs foraged, δ¹³C values in potential foods showed that food webs in subestuaries subject to higher N loads were supported by algal producers, while food webs in subestuaries with lower N loads were also supported by Spartina. δ¹³C values in horseshoe crab tissue did not change with load, suggesting they ate a mixed diet, regardless of N load. N and C isotopes in horseshoe crab feces were similar to signatures of estimated diet, suggesting low assimilation efficiency, perhaps due to ingestion of low quality organic matter. Although horseshoe crabs were relatively opportunistic in foraging habits, conservation or culture of horseshoe crabs may require habitats with higher water quality, ample particulate organic matter, and supporting a variety of prey.     

Phylogenetic relationship among horseshoe crab species: effect of substitution models on phylogenetic analyses

The horseshoe crabs, known as living fossils, have maintained their morphology almost unchanged for the past 150 million years. The little morphological differentiation among horseshoe crab lineages has resulted in substantial controversy concerning the phylogenetic relationship among the extant species of horseshoe crabs, especially among the three species in the Indo-Pacific region. Previous studies suggest that the three species constitute a phylogenetically unresolvable trichotomy, the result of a cladogenetic process leading to the formation of all three Indo-Pacific species in a short geological time. Data from two mitochondrial genes (for 16S ribosomal rRNA and cytochrome oxidase subunit I) and one nuclear gene (for coagulogen) in the four species of horseshoe crabs and outgroup species were used in a phylogenetic analysis with various substitution models. All three genes yield the same tree topology, with Tachypleus-gigas and Carcinoscorpius-rotundicauda grouped together as a monophyletic taxon. This topology is significantly better than all the alternatives when evaluated with the RELL (resampling estimated log-likelihood) method.     

Conservation education program for threatened Asian horseshoe crabs: A step towards reducing community apathy to environmental conservation

Long-term and locally-based public education programs are essential in supporting conservation initiatives for horseshoe crab protection, particularly in Asia, where their populations have been largely exploited for food and biomedical applications. However, the implementation of such conservation initiatives can be difficult due to a community’s ingrained apathy towards the environment. In addition, the effectiveness of similar education programs to engage the community in conservation work has rarely been assessed. We described the implementation strategies of a conservation education program in Hong Kong, with the objective to improve the community’s awareness, engagement and environmental attitudes towards the conservation of Asian horseshoe crabs. This study evaluated the program’s outreach, outcomes and impacts between 2009 and 2016. By raising laboratory-cultured juvenile horseshoe crabs at local secondary schools, this long-term conservation education program succeeded in enhancing students’ knowledge on the biology and ecology of horseshoe crabs, as well as promoting positive attitudes and behaviors towards relevant conservation issues. Lessons and recommendations derived from this program can serve as guidance for similar campaigns in other places.     

The Impact of Biomedical Bleeding on Horseshoe Crab, Limulus polyphemus , Movement Patterns on Cape Cod, Massachusetts

The purpose of this study was to determine if bleeding, for biomedical purposes, influenced the behavior of horseshoe crabs, Limulus polyphemus . In the summer of 2001, ten bled and ten control (unbled) female horseshoe crabs were tracked for 26 days using acoustic telemetry in a small estuary on Cape Cod, Massachusetts. All but three crabs, two bled and one control, were located during the study period. No mortality was observed in the control group, while 20% mortality was observed within the bled group. There was no significant difference in the average rate of movement or in the spatial distribution within the estuary between the two groups. However, a difference was detected in the movement patterns. Horseshoe crabs from the bled group had a random direction of movement compared to the directional movement pattern of the control group, suggesting that the bled crabs experienced more disorientation.     

A review on fisheries and conservation status of Asian horseshoe crabs

Horseshoe crabs are the only extant xiphosurans and are believed to be morphologically unchanged for more than 200 million years. Of the four extant species namely, Limulus polyphemus, Tachypleus tridentatus, Tapinauchenius gigas and Carcinoscorpius rotundicauda, the latter three are found in Asian waters. Recent evidences showed that Asian horseshoe crabs are facing serious threats such as degradation of their spawning grounds and habitat, environmental pollution, overexploitation as a culinary delicacy and biomedical bleeding practices. Baseline data on the distribution and existing population of the wild horseshoe crabs remain poorly known in several Asian regions. Several studies have clearly revealed that pressure due to over-fishing of wild stock has increased tremendously in the last decade. Due to an increase in demand for Tachypleus Amebocyte Lysate (TAL) analogous to Limulus Amebocyte Lysate (LAL) in the United States, there is an urgent need to comprehensively address their fishing and conservation measures in the Asian region. This review addresses the overall studies on three species of Asian horseshoe crabs in relation to their fishing practices, local exploitation of their wild stock either for human consumption (or) by biomedical industries. The authors have structured the discussion on an international scale to address the existing problems in fishing and conservation of horseshoe crabs. Since no specific regulatory force or legislative protection act or a policy to preserve their natural stock are available to this date, this paper strongly recommends representative countries to include horseshoe crabs under their wildlife protection act to avoid further unsustainable exploitation of their wild populations.     

Estimation of total hemolymph volume in the horseshoe crab Limulus polyphemus

Biomedical companies extract blood from the horseshoe crab, Limulus polyphemus, for the production of Limulus Amebocyte Lysate, used worldwide for detecting endotoxins in injectable solutions and medical devices. Despite the extensive use of horseshoe crabs by the biomedical industry, total hemolymph volume for this species is not known. The hemolymph volume of 60 adult horseshoe crabs was estimated using an inulin dilution technique. Blood volume of the horseshoe crab represented as a percentage of wet body weight was 25?±?2.2% for males and 25?±?5.1% (mean?±?SD) for females. Relationships between hemolymph volume and weight (p?=?0.0026, r ²?=?0.8762), hemolymph volume and prosomal width (p?<?0.0001), and hemolymph volume and inter-ocular width (p?<?0.0001) were observed. No significant differences were observed between males and females. The relationship of animal size and hemolymph volume can be used to predict how much blood can be drawn from horseshoe crabs used by the biomedical industry, and can be of further use in future bleeding mortality studies.     

Induction of Secondary Embryos by Intra- and Interspecific Grafts of Center Cells under the blastopore in Horseshoe Crabs

When the cell mass (center cells) of the early gastrulae in both American and Asian horseshoe crabs was grafted into the embryo of the homologous species, secondary embryos were formed as a result of these grafts. Secondary embryos were also formed in a similar way when the center cells of heterologous embryos were grafted between the American and Asian horseshoe crab embryos. The characteristics of the secondary embryos were similar to the host embryos in both cases, indicating that the center cells played the roles same as those by the amphibian organizer. The homogenate of center cells also induced the formation of secondary embryos. This is the first published study in which secondary embryos of horseshoe crabs have been induced by grafting. The fact may mean that this type of embryonic induction is widespread in the animal kingdom.     

Chelicerate Skeletal Neuromuscular Systems

In the past decade several laboratories began investigatingthe neuromuscular physiology of the arthropod subphylum, Chelicerata.This paper concerning chelicerate skeletal neuromuscular systemsconcentrates on three major areas: fine structural featuresof the skeletal muscles; fine structure of synaptic areas; andinnervation patterns and neuromuscular physiology. For all cheliceratesinvestigated, the fine structural features of the muscle fibersappear to be similar with the exception that the sarcomere lengthsfall into two general size categories (ca. 4 µ. and 7µ.) . Two types of synaptic areas are found in the chelicerates.In horseshoe crabs the synaptic region is formed in a largeevagination of the sarcoplasm, and evaginations from more thanone muscle fiber may combine into one synaptic region. In thearachnids the synaptic area is formed in a slight furrow inthe muscle fiber or along the surface of the fiber. Studiesof innervalion patterns reveal that horseshoe crabs have a muchgreater number of motor axons innervating the skeletal musculaturethan do other chelicerates. The motor axons appear to be similarin function in horseshoe crabs, but in scorpions there are atleast two functional types. In only one investigation has peripheralinhibition been demonstrated in Chelicerata.