牙鲆肠淋巴样组织免疫细胞研究Ⅰ：颗粒细胞的超微结构*

本文探讨了电镜下牙鲆(Paralichthys olivaceus)肠淋巴样组织内颗粒细胞的形态特点及其变化过程。颗粒细胞经鉴定为嗜酸性颗粒细胞，常存在于肠黏膜层及黏膜下层靠近肌肉层的淋巴腔中，以具有大型非匀质颗粒为其主要特征。牙鲆肠淋巴样组织内嗜酸性颗粒细胞的变化可分为增长期、成熟期、分泌期和衰退期四个时期，嗜酸性颗粒细胞具有明显的外排现象说明肠淋巴样组织是鱼类免疫防御系统的重要组成部分。     

光裸星虫体腔液细胞的显微和超微结构

利用光镜和透射电镜技术研究了光裸星虫(Sipunculus nudus)体腔液细胞(不包括生殖细胞)的显微和超微结构。结果表明,光裸星虫体腔液细胞密度为(1.73±0.51)×108/mL,可区分为8种类型,包括红细胞、嗜碱性粒细胞、嗜酸性颗粒细胞、淋巴样细胞、血栓细胞、网状细胞、盘状细胞和壶状细胞复合体,各类型细胞占细胞总数的比例分别为79.62%、5.23%、1.31%、2.86%、6.77%、1.74%、1.52%和0.94%。各类型细胞中,红细胞、嗜碱性粒细胞、嗜酸性颗粒细胞、淋巴样细胞、血栓细胞与其他无脊椎动物血液(体液)中的同类细胞类似。网状细胞、盘状细胞和壶状细胞复合体较为特殊。其中,网状细胞内有很多单层膜包被的空泡,大部分泡内电子密度很低,少数为中等电子密度;部分细胞内具有中间电子密度高、边缘电子密度低的颗粒;网状细胞可能是颗粒细胞释放颗粒后的残余体。盘状细胞表面密布微绒毛,胞质中有1个很大的液泡,液泡内容物电子密度低,有很多细而短的纤维状结构,该细胞的功能尚不了解。壶状细胞复合体由前端的泡状细胞和后端的纤毛细胞构成,能在体腔液中自由"游动",并能分泌粘液黏附各种衰老细胞、外来异物。     

中国对虾类淋巴器结构观察及功能探讨

中国对虾类淋巴器位于肝胰腺前方,左右各一个,呈白色,半透明状。它由触角动脉上分出的动脉血管不断分叉、迂曲,形成了一个外包透明结缔组织膜,内部充满小管的组织结构。类淋巴器的小管依其发育时期和功能不同表现为３种不同的形态。其中,第二种小管管壁具有生成和释放血细胞的功能。在类淋巴器中,观察到小颗粒细胞,大颗粒细胞和无颗粒细胞。小颗粒细胞具有吞噬功能,在受到抗原刺激时可以胞吐方式释放细胞中的电子致密颗粒。大颗粒细胞无吞噬功能。无颗粒细胞又可分为巨噬细胞样无颗粒细胞和淋巴细胞样无颗粒细胞。前者具有很强的吞噬能力,而后者在受到抗原刺激时,可表现出类似浆细胞样分化的细胞特征。     

斑鱾消化系统形态学与组织学研究

为探究斑鱾(Girella punctata Grayy)的消化生理特性,本实验采用形态学与组织学方法对斑鱾消化系统主要器官及组织(口咽腔、食道、胃、肠及肝脏)进行研究,并分析其组织结构与功能的关系。结果显示：食道腔壁的组织结构从内到外依次为黏膜层、黏膜下层、肌层和浆膜层,这与大部分硬骨鱼类相似。胃为U型胃,主要由贲门部、胃体部和幽门部三部分组成。黏膜层和黏膜下层向胃腔突起形成8个突起皱壁,黏膜层内有丰富的腺体,消化管内壁皱褶多,肌层厚;肠道分为前肠、中肠和后肠,其比肠长约1.54±0.06,腔壁向内形成6个不规则的褶皱突起;肝脏由实质和间质两部分组成,分为左右两叶。斑鱾的消化系统结构特征与其杂食性相适应。     

中国对虾体内一种结缔组织颗粒细胞的电镜观察和组化研究

运用半薄切片、超薄切片和组织细胞化学技术，在光镜和电镜下，观察研究中国对虾中肠结缔组织中的1种颗粒细胞，该细胞28～32μm大小，细胞核小，胞质颗粒呈多样性。亚甲基兰染色显示，细胞内颗粒具异染性，提示这些颗粒中含有肝素类成分；磷钨酸乙醇法染色显示，该细胞内有嗜铬颗粒并具有肥大细胞颗粒的染色性质；焦锑酸甲法显示，细胞中富含Ca^2 ；酚氧化酶孵育反应显示。酶活性出现在受病毒感染对虾肠壁结缔组织中的这种颗粒细胞中。研究表明：1．存在于中肠背侧结缔组织中的颗粒细胞，在分布位置、细胞形态和相关染色反应结果等方面不同于已往报道的血细胞，这是1种具有某些肥大细胞特征的结缔组织颗粒细胞；2．结缔组织颗粒细胞具有储存Ca^2 的功能；3．病毒感染，可使结缔组织颗粒细胞中的酚氧化酶原转化成有活性的酚氧化酶，提示这种细胞参与对虾的免疫防御反应。     

黑鱾消化系统的形态学及组织学研究

鱼类消化系统的组织结构特点与食性密切相关。为了探究岛礁性鱼类黑魢的消化生理特点,文章以黑鱾(Girella leonina)为研究对象,采用苏木精-伊红(H.E)染色法研究其消化系统的组织结构特点,探讨其组织结构特点与食性、环境适应性之间的相关性。结果显示:1)食道向腔内形成纵向6个粗大皱褶,该皱褶具明显横纹肌,扩张性较强,黏膜上皮间分布着黏液细胞,呈锥形;2)黑鱾为"V"形胃,可分为贲门部、胃体部和幽门部3个部分,胃腺横切面为圆形,管腔由数个腺细胞围成,胃肌层平滑肌层极厚,为内环肌层外纵肌层结构,其中胃体部固有层处胃腺最为发达,胃黏膜形成许多皱襞,幽门胃具多级分枝皱褶;3)幽门处分布有150多条分枝状盲囊,盲囊腔内充满黏膜褶,黏膜上皮由单层柱状上皮细胞组成,不具有杯状细胞;4)肝脏分为左右两叶,组织结构呈现出一般硬骨鱼类的共性,肝细胞体积较大,呈多边形,大小约为30μm;5)肠道具有4个肠曲,比肠长值约为1.52±0.06,介于植食性和肉食性鱼类之间,整个肠道均有大量杯状细胞分布,肠道内具发达的小肠绒毛凸起,绒毛基部上皮向固有层内陷形成肠腺,腺体开口于相邻绒毛基部之间,前中肠作为主要消化吸收场所,其绒毛数量和高度要大于后肠。文章研究了黑鱾消化系统组织学特征与其消化、吸收的关系,认为黑鱾消化系统具备典型杂食性鱼类消化系统的特征。     

黑鱾消化系统的形态学及组织学研究*

鱼类消化系统的组织结构特点与食性密切相关。为了探究岛礁性鱼类黑魢的消化生理特点, 文章以黑鱾(Girella leonina)为研究对象, 采用苏木精-伊红(H.E)染色法研究其消化系统的组织结构特点, 探讨其组织结构特点与食性、环境适应性之间的相关性。结果显示: 1) 食道向腔内形成纵向6个粗大皱褶, 该皱褶具明显横纹肌, 扩张性较强, 黏膜上皮间分布着黏液细胞, 呈锥形; 2) 黑鱾为“V”形胃, 可分为贲门部、胃体部和幽门部3个部分, 胃腺横切面为圆形, 管腔由数个腺细胞围成, 胃肌层平滑肌层极厚, 为内环肌层外纵肌层结构, 其中胃体部固有层处胃腺最为发达, 胃黏膜形成许多皱襞, 幽门胃具多级分枝皱褶; 3) 幽门处分布有150多条分枝状盲囊, 盲囊腔内充满黏膜褶, 黏膜上皮由单层柱状上皮细胞组成, 不具有杯状细胞; 4) 肝脏分为左右两叶, 组织结构呈现出一般硬骨鱼类的共性, 肝细胞体积较大, 呈多边形, 大小约为30μm; 5) 肠道具有4个肠曲, 比肠长值约为1.52±0.06, 介于植食性和肉食性鱼类之间, 整个肠道均有大量杯状细胞分布, 肠道内具发达的小肠绒毛凸起, 绒毛基部上皮向固有层内陷形成肠腺, 腺体开口于相邻绒毛基部之间, 前中肠作为主要消化吸收场所, 其绒毛数量和高度要大于后肠。文章研究了黑鱾消化系统组织学特征与其消化、吸收的关系, 认为黑鱾消化系统具备典型杂食性鱼类消化系统的特征。     

波纹唇鱼消化道的形态学和组织学

通过形态解剖、组织切片和光镜观察技术对波纹唇鱼Cheilinus undulatus消化道的形态学和组织学进行了研究。结果表明,波纹唇鱼消化道分为口咽腔、食道、胃和肠。口咽腔较大,颌齿和咽骨齿均发达,具有多种类型齿;食道粗而短;鳃耙仅有3对半,较长、粗且稀少;"I"型胃,盲囊部不明显;无幽门盲囊;肠管粗短,可分为前肠、后肠和直肠三部分,肠道系数约为3.05。从食道后部开始整个消化道管腔有密集的绒毛,消化道管壁的组织结构由黏膜层、黏膜下层、肌肉层和浆膜层构成。消化道上皮为单层柱状细胞,其间分布有较多杯状细胞,杯状细胞有明显的开口。黏膜下层为疏松结缔组织。肌肉层致密,内层为环肌外层为纵肌,二者间分布有神经丛。浆膜层极薄,覆盖于整个消化道的外层。这些特征使波纹唇鱼具有典型的肉食性鱼类的特征和习性,便于咬破和磨碎食物坚硬的外壳,使食物在消化道中停留的时间较短、能够快速通过消化道,在肠道中迅速消化吸收,然后排出体外。     

海湾扇贝血细胞形态学比较研究

为给海湾扇贝的细胞免疫学研究提供基本依据,同时采用光镜和电镜技术观察海湾扇贝血细胞的形态特征,并进行血细胞酸性磷酸酶活性的初步分析.结果表明,海湾扇贝的血细胞可分为透明细胞和颗粒细胞2种类型.与透明细胞相比,颗粒细胞数目多,酸性磷酸酶活性高,细胞核呈肾形或卵圆形,核质比低,细胞质内颗粒状物质丰富,颗粒大小不一,可明显分为大颗粒细胞和小颗粒细胞2类.这些特征表明,海湾扇贝颗粒细胞和透明细胞的免疫学功能是明显不同的.     

嫁■消化系统的组织学研究

利用光学显微镜研究了嫁(Cellana toreuma(Reeve))消化系统的组织结构。嫁的消化系统由消化道(口区、食道、嗉囊、胃、肠、直肠、肛门)和消化腺(食道腺、肝胰腺)组成。口中具有齿舌和颚片,分别通过发达的肌肉与软骨相连;食道中部膨大成食道腺;胃和肠非常长。消化道壁由黏膜层、黏膜下层、肌层和外膜构成,黏膜上皮主要为单层柱状上皮细胞,肌层主要为环肌。胃壁和肠壁非常薄,黏膜下层和外膜均不发达。肝胰腺由无数肝小叶组成,每一肝小叶由位于基膜上的单层腺细胞和胚细胞组成。     

牙鲆肠淋巴样组织免疫细胞研究Ⅱ.无颗粒白细胞的超微结构

运用电镜技术,在牙鲆肠淋巴样组织鉴定出４种无颗粒白细胞,即淋巴细胞、巨噬细胞、单核细胞和浆细胞样细胞。它们的形态均比在造血器官中多样化。巨噬细胞形态极其有规则;单核细胞的细胞质中有较多空泡;淋巴细胞具多且细长的伪足,而浆细胞样细胞是以内含大量的粗面内质网为其主要特征。同时,作者对无颗粒白细胞的形态与功能之间的关系进行了探讨。     

北冰洋海单胞菌β-D-半乳糖苷酶的异源表达及酶学特性研究

初筛表明,一株分离自北极加拿大海盆海冰心芯样品的海单胞菌(Marinomonas sp.BSi20584)具有较高的β-D-半乳糖苷酶活性,为了研究清楚其酶学性质,将经hiTAIL-PCR扩增得到的β-D-半乳糖苷酶基因(galt)与pET-28a(+)原核表达载体结合,转入大肠杆菌BL21(DE3)。经IPTG诱导后对重组β-D-半乳糖苷酶(GALT)的表达条件进行了优化,采用金属螯合亲和层析技术制备纯酶,并对重组GALT的酶学性质进行了研究。结果显示,重组酶的最适诱导温度为20℃,在IPTG浓度为0.07mmol/L时诱导22h后,酶活和产酶量达到最大值。GALT单体分子量约为6.6×104 g/mol,天然酶为同源三聚体。GALT最适作用温度为35℃,其热稳定性较好,在60℃处理5h后,仍可保持50%以上的相对活性。GALT的最适作用pH为9.0,在pH为6.0~11.0范围内比较稳定。GALT的最适NaCl浓度为0.5mol/L,对盐度具有较高的耐受性。Mg2+、K+、DTT和EDTA对酶活不具有显著影响,而Mn2+、Fe2+对酶活有促进作用,Zn2+和L-谷胱甘肽对酶活有抑制作用。GALT对Galβ1-4GlcNAc具有水解作用,而对Galβ1-3GalNAc和Galβ1-3GlcNAc糖苷键型没有水解能力。本研究实现了海单胞菌属菌株的β-D-半乳糖苷酶基因在大肠杆菌系统中的高效表达,并系统研究了重组酶的酶学特性,为后续开展该酶的代谢适应性和潜在应用研究提供详细的酶学数据基础。     

Experimental studies on dimethylsulfide (DMS) and dimethylsulfoniopropionate (DMSP) production by four marine microalgae

The production of dimethylsulfide (DMS) and dimethylsulfoniopropionate (DMSP) by marine microalgae was investigated to elucidate more on the role of marine phytoplankton in ocean-atmosphere interactions in the global biogeochemical sulfur cycle.Axenic laboratory cultures of four marine microalgae–Isochrysis galbana 8701,Pavlova viridis,Platymonas sp.and Chlorella were tested for DMSP production and conversion into DMS.Among these four microalgae,Isochrysis galbana 8701 and Pavlova viridis are two species of Haptophyta,while Chlorella and Platymonas sp.belong to Chlorophyta.The results demonstrate that the four algae can produce various amounts of DMS(P),and their DMS(P) production was species specific.With similar cell size,more DMS was released by Haptophyta than that by Chlorophyta.DMS and dissolved DMSP (DMSPd) concentrations in algal cultures varied significantly during their life cycles.The highest release of DMS appeared in the senescent period for all the four algae.Variations in DMSP concentrations were in strong compliance with variations in algal cell densities during the growing period.A highly significant correlation was observed between the DMS and DMSPd concentrations in algal cultures,and there was a time lag for the variation trend of the DMS concentrations as compared with that of the DMSPd.The consistency of variation patterns of DMS and DMSPd implies that the DMSPd produced by phytoplankton cells has a marked effect on the production of DMS.In the present study,the authors’ results specify the significant contribution of the marine phytoplankton to DMS(P) production and the importance of biological control of DMS concentrations in oceanic water.     

Winter distribution of euphausiids (Euphausiacea) in the Barents Sea (2000–2005)

The purpose of the study is to analyze the state of the Barents Sea euphausiids populations in the warm period (2000–2005) based on the study of their structure dynamics and distribution under the influence of abiotic and biotic factors. For estimation of their aggregations in the bottom layer, the traditional method was used with the help of the modified egg net (0.2 m² opening area, 564 μm mesh size). The net is used for collecting euphausiids in the autumn–winter period when their activity is reduced, which results in high-catch efficiency. The findings confirmed the major formation patterns of the euphausiids species composition associated with climate change in the Arctic basin. As before, in the warm years, one can see a clear-cut differentiation of space distribution of the dominant euphausiids Thysanoessa genus with localization of the more thermophilic Thysanoessa inermis in the north-west Barents Sea and Thysanoessa raschii in the east. The major euphausiids aggregations are formed of these species. In 2004, the first data of euphausiids distribution in the northern Barents Sea (77–79°N) were obtained, and demonstrated extremely high concentrations of T. inermis in this area, with the biomass as high as 1.7–2.4 g m⁻² in terms of dry weight. These data have improved our knowledge of the distribution and euphausiids abundance during periods of elevated sea-water temperatures in the Barents Sea. The oceanic Atlantic species were found to increase in abundance due to elevated advection to the Barents Sea during the study period. Thus, after nearly a 30-year-long absence of the moderate subtropical Nematoscelis megalops in the Barents Sea, they were found again in 2003–2005. However in comparison with 1960, the north-east border of its distribution considerably shifted to 73°50′N 50°22′E. The portion of Meganyctiphanes norvegica also varied considerably—from 10% to 20% of the total euphausiids population in the warm 1950s–1960s almost to complete disappearing in 1970–1990s. The peak of this species’ occurrence (18–26%) took place in the beginning of warm period (1999–2000) after a succession of cold years. The subsequent reduction of the relative abundance of M. norvegica to 7% might have been mostly caused by fish predation during a period of low population densities of capelin. This high predation pressure may therefore have been mediated both by other pelagic fishes (i.e. herring, blue whiting, polar cod) but also by demersal fishes such as cod and haddock. Similar sharp fluctuations in the capelin stock (the major consumer of euphausiids) created marked perturbations in the food web in the Barents Sea in the middle 1980s and the early 1990s.     

黑斑口虾蛄(Oratosquilla Kempi Schmitt)配子发生的显微观察

观察黑斑口虾蛄Oratosquillakempi的精巢和卵巢 ,并对精、卵巢进行组织切片显微观察。结果表明 :黑斑口虾蛄的精巢由 1对很细的管状结构组成 ,精子发育可分为精原细胞、初级精母细胞、次级精母细胞、精子等期 ,精子圆球形 ,无鞭毛。卵巢分左、右两叶 ,对称且相互粘连。卵细胞为多黄卵 ,卵母细胞发育分为未发育期、卵黄形成前期、卵黄形成期和成熟期 4期 ;在卵黄形成前期形成大的生发泡。卵巢发育分为未发育期、发育早期、发育期、成熟前期、成熟期和产卵后期 6期。     

Thermal marking of rainbow trout (Oncorhynchus mykiss) otoliths

A small‐scale experiment was done to test the feasibility of thermally marking hatchery‐reared rainbow trout (Oncorhynchus mykiss Walbaum) fry that are released into rivers and impoundments in New South Wales (NSW), Australia. Fry of rainbow trout were exposed to two different 48‐h thermal cycles each of a cold and warm water period. One thermal regime consisted of a cold water period during which the temperature was reduced from 14 to 8°C for 18 h followed by a return to 14°C for 30 h. For the second thermal treatment, water temperature was reduced to 4°C for 10 h followed by a period of 38 h at 14°C. Thermal cycles were repeated 4 and 8 times for each thermal regime, respectively. Following a growth period after treatment, obvious marks were visible on all treated otoliths as distinct from control otoliths. The 10°C differential treatment created the most visible patterns and growth of these fish was not significantly different from control fish. This marking method could be applied to normal hatchery practices to evaluate the effectiveness of large‐scale rainbow trout stockings in NSW.     

Population dynamics of Callichirus major (Say, 1818) (Crustacea, Thalassinidea) on a beach in northeastern Brazil

We describe the structure, reproductive cycle, fecundity, growth, and mortality of a harvested population of the ghost shrimp Callichirus major. Samples were collected at monthly intervals from September 1999 to October 2000 on an urban sandy beach (08°11′S 34°55′W) in northeastern Brazil. During this period the sex ratio did not differ significantly from 1:1 (0.98 M: 1 F). Minimum and maximum sizes of the Dorsal Oval were 2.59 and 12.19 mm for males and 4.46 and 12.62 mm for females, respectively. Ovigerous females were found throughout the period, except between August and September 2000. Maximum lifespan was estimated as 3.3 and 3.4 years for females and males, respectively. This northeastern population differed from others previously studied in southern and southeastern Brazil, in regard to sex ratio, maximum attained size, maturation size, period and duration of the reproductive cycle, and fecundity. We interpret these regional differences as evidence for over-fishing at the study site, and suggest that large-scale management plans for callianassid populations should use regional population parameters.     

水母(沙海蜇)降解与营养盐(氮、磷)释放的模拟与实证

The aim of this study was to investigate nitrogen and phosphorus released in the process of the decomposition of giant jellyfish in the laboratory and found the evidence to verify the influence of nutrients released by the decomposition of jellyfish on the ecosystem in the field. The release of nitrogen and phosphorus from the decomposition of Nemopilema nomurai was examined in a series of experiments under different incubation conditions such as different p H values, salinity values, temperatures and nitrogen and phosphorus concentrations. The results showed that the complete decomposition of Nemopilema nomurai generally took about 4–8 d. The release of nitrogen and phosphorus from the decomposition of Nemopilema nomurai could be divided into two stages: the early stage and the later stage, although the efflux rate of nitrogen was one order more than phosphorus. In the early stage of the decomposition of Nemopilema nomurai, the concentrations of dissolved nitrogen, dissolved phosphorus, total nitrogen and total phosphorus in seawater increased rapidly, and the concentration of nitrogen could reach the highest level in the whole degradation process. In the later stage of the decomposition, the concentrations of dissolved nitrogen and total nitrogen declined slowly, while the concentration of phosphorus in water could reach a maximum in the degradation process. High p H, low salinity,high temperature and N/P will promote the release of nitrogen; low p H is unfavorable to the release of nitrogen but favorable to the release of phosphorus. In addition, we found the concentrations of ammonium and phosphate in the bottom water were higher than those in the surface water during the period of jellyfish bloom in the Jiaozhou Bay, proving that nutrients released by the decomposition of jellyfish have significant influence on nitrogen and phosphorus in the field. For the whole Yellow Sea, nutrients released by jellyfish carcasses may reach up to(2.63±2.98)×107 mol/d of dissolved nitrogen(DN) and(0.74±0.84)×106 mol/d of dissolved phosphorus(DP) during the period of jellyfish bloom. The values are comparable to riverine inputs in a day, but much higher than sediment–water exchange flux in the Yellow Sea. The great amounts of nutrients must have significant influence on the nutrients balance of the Yellow Sea during the period of jellyfish dead and decomposition. Both the experimental data and field observations proved that the decomposition of jellyfish may release a great amount of nutrient to the surrounding environment during the period of jellyfish decomposition.     

Temporal dynamics of shallow subtidal meiobenthos from a beach in Tenerife (Canary Islands, northeast Atlantic Ocean)

A shallow subtidal (3 m deep) meiofaunal assemblage in Los Abrigos Bay, Tenerife, Canary Islands was sampled during May 2000-April 2001. Themain aims were to (1) find temporal variations inmeiofaunal assemblage structure and overall abundance, as well as in the most abundant meiofaunal species throughout the study period, and (2) identify environmental variables (sedimentary composition, organic matter content, and total nitrogen) that better explain meiofaunal assemblage structure during the study year. The most abundant species were the nematodes Siphonolaimus sp. 2 and Catanema sp., which represented 46.2% of the overall meiofaunal abundances and varied significantly throughout the study duration. Overall meiofaunal abundance and the most abundant taxonomic groups (nematodes, copepods, and oligochaetes) showed significant temporal variations during the study period. Nematodes overwhelmingly dominated during the study period, ranging from 78% in May to 97.34% in February. Multivariate analyses showed seasonality inmeiofaunal community structure during the study year, with the lowest abundances in May.