共查询到19条相似文献，搜索用时 140 毫秒
1.
流式细胞术在克氏原螯虾血细胞的分类、活性和免疫功能研究中的应用 总被引：2，自引：0，他引：2
应用流式细胞术(FCM)对克氏原螯虾(Procambarus clarkii)血细胞的分类、活性和免疫功能进行了研究。结果显示:血细胞可分为透明细胞、小颗粒细胞和大颗粒细胞三个亚群,组成比例分别为(26.25±5.29)%、(51.44±7.02)%和(11.20±1.82)%;螯虾血细胞的平均总凋亡率约为3.12%;血细胞对荧光大肠杆菌的吞噬活力显著(P<0.05)高于荧光微球,吞噬率分别为17.04%和14.57%;血细胞在自然生理状态下含有一定量的活性氧,其在两类颗粒细胞的含量显著(P<0.05)高于透明细胞,在大颗粒细胞中最高。结果表明,FCM能较好地应用于虾类的血细胞分类和功能研究。 相似文献
2.
中华绒螯蟹的血细胞组成、分类及免疫学功能 总被引：1，自引：1，他引：0
为了解中华绒螯蟹血细胞组成、分类及其在免疫应答过程中起到的重要作用,本研究通过细胞化学和细胞酶学分析,并结合细胞形态观察,对中华绒螯蟹血细胞的组成、分类进行了研究。同时,通过人工感染嗜水气单胞菌后血液的总血细胞数(THC)和不同类型血细胞数(DHC)的变化,研究不同类型血细胞在免疫应答过程中所发挥的作用。结果显示,依据本实验分类方法,中华绒螯蟹血细胞可以分为4类:大颗粒细胞(G)、中间型颗粒细胞(IG)、小颗粒细胞(SG)和透明细胞(H)。其中小颗粒细胞数量最多,约占33.54%±0.98%,中间型颗粒细胞最少,仅占15.31%±2.01%。4种类型细胞中均含有多糖成分而不含脂质,只有大颗粒细胞在酸性磷酸酶、碱性磷酸酶、β-葡萄糖醛酸酶以及酚氧化酶染色中发现阳性反应。此外,嗜水气单胞菌感染后中华绒螯蟹总血细胞数在3 h后明显升高,在6 h达到峰值,约9.57×106个/m L,并显著高于未处理组和生理组;在感染过程中,大颗粒细胞数量明显下降而透明细胞数量明显上升,这种现象在6 h时达到顶峰并随着时间延长而逐渐恢复至正常水平。研究表明,在中华绒螯蟹非特异性免疫应答过程中,透明细胞主要通过大量增殖执行吞噬功能来参与免疫应答,而大颗粒细胞主要通过裂解释放胞质中所含免疫相关酶参与免疫反应。其中,各类型细胞之间可能存在相互转化作用,而中间型颗粒细胞为这种转化中的过渡类型。 相似文献
3.
三种模式开展克氏原螯虾人工增养殖试验 总被引：1，自引：0，他引：1
克氏原螯虾(Procambarus clarkia)属甲壳动物纲、软甲亚纲、十足目、螯虾科,俗称小龙虾,杂食陛,繁殖力强,适应性强,生长快,抗病力强,成活率高,近年来不仅在国内成为畅销的水产品,而且其虾仁、虾黄及整肢虾出口亦迅速增加,呈现供不应求的趋势.为探索适合湖区克氏原螯虾增养殖技术,满足市场需求,笔者近年开展了克氏原螯虾人工增养殖试验,有关结果整理如下,以备参考. 相似文献
4.
白洋淀日本沼虾血细胞的初步研究 总被引：2，自引：0，他引：2
对取自华北白洋淀种群的日本沼虾(Macrobrachium nipponense)血细胞进行显微形态观察、分类、测量和计数。依据细胞的形态,胞质中颗粒的数量和密度以及细胞的核质比把日本沼虾血细胞分为透明细胞(hyaline cell,HC)、半颗粒细胞(semigranular cell,SGC)和颗粒细胞(granular cell,GC)3类。血细胞大小依次为颗粒细胞>半颗粒细胞>透明细胞。血细胞密度为(1.13±0.50)×106个/mL,其中GC占48.5%,SGC占31.8%,HC占19.7%。并对我国日本沼虾的华南种群、华中种群及白洋淀所在地华北种群的血细胞大小和数量进行了比较。 相似文献
5.
为了更精确地对甲壳动物血细胞进行快速分类和功能分析,本研究以中华绒螯蟹为例,依据其血细胞内具有颗粒结构的特征,与常规显微观察对比,探讨了一种基于图像流式细胞仪的血细胞自动化分类新方法,并测量了体质量为(10±3) g的中华绒螯蟹的血细胞对直径1μm微球的吞噬情况。结果显示,两种方法都可将中华绒螯蟹血细胞分为4个类群。显微镜观察分类基于胞内可见颗粒,但由于缺乏精细的量化标准,批次样品的人工辨识结果波动较大;而图像流式方法以胞内全部颗粒结构为对象,利用高精度检测模块测量,结果更准确、客观,而且通量高、重复性好。测量结果显示,中华绒螯蟹血淋巴中无颗粒、小颗粒、中颗粒及大颗粒细胞占比分别为40.62%±2.65%、36.68%±6.84%、7.80%±1.16%和16.51%±5.60%,依据测量的颗粒特征区分的4个类群界限清晰,缺乏过渡样点,提示各类细胞之间可能没有相互转化。进一步的活体微球吞噬实验证实,中华绒螯蟹的4类血细胞都具有吞噬功能,并以无颗粒细胞为主要吞噬类群;吞噬微球的细胞比例在注射后6 h内呈钟型曲线变化,4 h可达峰值(5.69%±0.44%),表明中华绒螯蟹血细胞能高效清除血淋巴中的异物。研究表明,图像流式细胞仪适合于中华绒螯蟹的血细胞分类分析和功能研究,本研究结果为同类研究提供了重要参考,将有助于更全面了解甲壳动物血细胞的功能。 相似文献
6.
流式细胞术与显微观察对罗氏沼虾血细胞的比较研究 总被引：2，自引：1，他引：1
应用流式细胞术和显微观察对罗氏沼虾（Macrobrachium rosenbergii）血细胞的分类分群和各类细胞组成比例进行研究,比较2种方法的优缺点。应用显微观察,根据细胞颗粒的有无和大小以及细胞染色情况,可把罗氏沼虾血细胞分为透明细胞、小颗粒细胞和大颗粒细胞。根据其血细胞的前向角散射光（FSC）和侧向角散射光（SSC）特性的不同,也将血细胞分为3个亚群：透明细胞、小颗粒细胞和大颗粒细胞。检测各种细胞所占的比例,2种方法测得比例均为：小颗粒细胞〉大颗粒细胞〉透明细胞;2方法的结果存在显著差异（P〈0．05）,但相关性极显著（P〈0．01）。利用流式细胞术可更快捷、准确,减少人为误差,显微观察可对细胞内部结构进行更深入的分析,并可进行细胞实际大小和细胞密度的测定。2种方法互补不足,可更快速、准确、全面地进行细胞基础特征的研究。 相似文献
7.
分析了克氏原螯虾Procambarus clarkia人工感染白斑综合征病毒(White Spot Syndrome Virus,WSSV)后,其血细胞吞噬(Phagocytosis)活性,肝胰腺酸性磷酸酶(ACP)、碱性磷酸酶(AKP)活性的变化规律,其目的是为WSSV感染与螯虾的免疫防御反应等研究提供依据.分析结果显示,随着WSSV感染克氏原螯虾时间的延长,血细胞吞噬活性、肝胰腺酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性也随之改变,其中血细胞吞噬活性的变化规律性比较明显,可作为WSSV感染的敏感指标,也可以用作间接指示克氏原螯虾健康状况的指标.而在不同的感染时间,克氏原螯虾肝胰腺ACP与AKP的活性波动较大,无明显的规律可循,难以作为WSSV感染的敏感指标. 相似文献
8.
为研究虾蟹类甲壳动物血细胞膜表面是否具有相同的抗原决定簇及共同抗原表位的生物学特征,采用制备的抗三疣梭子蟹颗粒血细胞单克隆抗体,通过激光共聚焦扫描显微镜(laser scanning confocal microscopy,LSCM)、免疫印迹(Western blot)和流式细胞术(Flow Cytometry,FCM)等多种方法,测定了7种甲壳类动物(凡纳滨对虾、中国明对虾、刀额新对虾、口虾蛄、中华绒螯蟹、日本板蟹及美洲黄道蟹)颗粒血细胞及透明血细胞与三疣梭子蟹颗粒血细胞单克隆抗体发生特异性结合的抗原表位。LSCM可观察到该株抗三疣梭子蟹颗粒血细胞单克隆抗体与中华绒螯蟹和日本板蟹血细胞交叉反应结果为阳性,分析发现在中华绒螯蟹血淋巴中阳性颗粒血细胞在其颗粒血细胞总数中占76.74%,阳性透明血细胞在其透明血细胞总数中占70.59%,日本板蟹阳性颗粒血细胞在其颗粒血细胞总数中占73.86%,阳性透明血细胞在其透明血细胞总数中占16.67%,其余5种甲壳类动物均为阴性;Western blot测试结果显示该株单克隆抗体仅与中华绒螯蟹血细胞反应,且发生反应的抗原决定簇位于分子量为30 ku的蛋白带上;FCM分析发现该株单克隆抗体与中华绒螯蟹透明血细胞和颗粒血细胞均可发生交叉反应,阳性率分别为57.72%和77.05%,与日本板蟹透明血细胞阳性反应极少,阳性率仅为9.57%,与颗粒血细胞发生阳性反应的阳性率为82.59%。 相似文献
9.
以投喂患白斑综合征病毒病(WSSV)的南美白对虾组织的方法人工感染健康的克氏原螯虾,利用PCR检测实验克氏原螯虾的感染情况;同时比较健康的克氏原螯虾和感染病毒的克氏原螯虾体内ATPase活性.结果显示:白斑综合征病毒可以感染克氏原螯虾,受感染的克氏原螯虾体内病毒含量与其死亡呈正相关;健康的克氏原螯虾和感染病毒的克氏原螯虾体内的ATPase活性分别为7.337 U/mgprot和4.212 U/mgprot,有极其显著的差异. 相似文献
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11.
采用光镜和电镜的方法,以血细胞所含颗粒与否,颗粒多少和核质比等将成年日本新糠虾血细胞进行分型,并对各型血细胞所占比例进行统计。利用生化分析的方法,对不同发育阶段(新孵后0、10、20和30d)日本新糠虾全组织中磷酸酶(酸性磷酸酶和碱性磷酸酶)、溶菌酶和酚氧化酶的活性进行测定。结果表明,日本新糠虾血细胞可为3种类型,它们分别为透明细胞、半颗粒细胞和颗粒细胞。它们的胞体大小依次增大,核质比依次降低。对此3种类型血细胞比例分别统计后发现,颗粒细胞所占比例最多约为47.53%±0.02,其次是半颗粒细胞约为31.23%±0.01,最少的是透明细胞约为21.24%±0.02。日本新糠虾体内磷酸酶(酸性磷酸酶和碱性磷酸酶)、溶菌酶和酚氧化酶的活性均有随个体发育而呈降低的趋势。3种酶中以溶菌酶的活性最高,平均约为305.20U/mg蛋白,其次是酸性磷酸酶约为0.2365U/mg蛋白,而酚氧化酶和酸性磷酸酶在成体中的活性基本一致,约为0.1295U/mg蛋白。研究表明,日本新糠虾血细胞组成中,以颗粒细胞为主。溶菌酶是其主要的免疫活性物质。日本新糠虾体内磷酸酶(酸性磷酸酶和碱性磷酸酶)、溶菌酶和酚氧化酶活性有随其个体发育而逐渐... 相似文献
12.
凡纳滨对虾白斑综合征血液病理研究 总被引：1，自引：0，他引：1
对自然发病、投喂和注射感染的凡纳滨对虾白斑综合征（white spot syndrome，WSS）血液病理进行研究，结果发现：不同感染方式患病对虾的血液病理变化相似，表现为：1．患病对虾血细胞总数、透明细胞数量极显著减少，小颗粒细胞、大颗粒细胞极显著增加。2．显微病理变化主要表现为血涂片中血细胞明显减少且分布不均匀，破损或解体的细胞增多，呈典型的溶血状态。3．超微病理变化表现为，大部分血细胞坏死，少数血细胞呈不典型的凋亡。患病对虾的血细胞核中可见大量白斑综合征病毒（white spot syndrome virus，WSSV）粒子。病理变化表明血细胞是WSSV的主要靶细胞。 相似文献
13.
Francisco Vargas-Albores Teresa Gollas-Galván & Jorge Hernández-López 《Aquaculture Research》2005,36(4):352-360
Haemocytes from penaeid shrimp (Farfantepenaeus californiensis, Litopenaeus vannamei and L. stylirostris) were separated using a discontinuous Percoll gradient. Shrimp haemocytes were spontaneously adhered to glass, allowing slide preparations for staining and microscopic differential counting. Like other crustaceans, shrimp has three main populations differing in presence and size of cytoplasmic granules and each population seems to be biochemical or functionally compromised. Prophenoloxidase (proPO )activity was mainly located in large granules haemocytes (75%) while the small granules cells participate with 25%, but seem to be responsible for encapsulation. Haemocyte discrimination ability was tested using Sephadex? (Seph?), DEAE‐Seph? and CM‐Seph?. Only DEAE‐Seph? was encapsulated by shrimp haemocytes and provoked the release of proPO activating system, indicating the role of particle charge in the activation of shrimp immune response. 相似文献
14.
Kinetic analysis of internalization of white spot syndrome virus by haemocyte subpopulations of penaeid shrimp,Litopenaeus vannamei (Boone), and the outcome for virus and cell 
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下载免费PDF全文 V V Tuan G M A De Gryse K V Thuong P Bossier H J Nauwynck 《Journal of fish diseases》2016,39(12):1477-1493
Little is known about the innate antiviral defence of shrimp haemocytes. In this context, the haemocytes of penaeid shrimp Litopenaeus vannamei (Boone) were separated by iodixanol density gradient centrifugation into five subpopulations (sub): sub 1 (hyalinocytes), sub 2 and 3 (prohyalinocytes), sub 4 (semigranulocytes) and sub 5 (granulocytes) and exposed to beads, white spot syndrome virus (WSSV) and ultraviolet (UV)‐killed WSSV. In a first experiment, the uptake of beads, white spot syndrome virus (WSSV) and UV‐killed WSSV by these different haemocyte subpopulations was investigated using confocal microscopy. Only haemocytes of sub 1, 4 and 5 were internalizing beads, WSSV and UV‐killed WSSV. Beads were engulfed by a much larger percentage of cells (91.2 in sub 1; 84.1 in sub 4 and 58.1 in sub 5) compared to WSSV (9.6 in sub 1; 10.5 in sub 4 and 7.9 in sub 5) and UV‐killed WSSV (12.9 in sub 1; 13.3 in sub 4; and 11.8 in sub 5). In a second experiment, it was shown that upon internalization, WSS virions lost their envelope most probably by fusion with the cellular membrane of the endosome (starting between 30 and 60 min post‐inoculation) and that afterwards the capsid started to become disintegrated (from 360 min post‐inoculation). Expression of new viral proteins was not observed. Incubation of haemocyte subpopulations with WSSV but not with UV‐killed WSSV and polystyrene beads resulted in a significant drop in haemocyte viability. To find the underlying mechanism, a third experiment was performed in which haemocyte subpopulations were exposed to a short WSSV DNA fragment (VP19) and CpG ODNs. These small DNA fragments induced cell death. In conclusion, WSSV is efficiently internalized by hyalinocytes, semigranulocytes and granulocytes, after which the virus loses its envelope; as soon as the capsids start to disintegrate, cell death is activated, which in part may be explained by the exposure of viral DNA to cellular‐sensing molecules. 相似文献
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Argopecten irradians is one of the most important commercial species of Pectinidae family in China. The internal defense system of mollusks consists of circulating haemocytes. In order to characterize the haemocytes of the scallop A. irradians, light and electron microscopical studies were carried out. Four types of haemocytes were recognized: type I small hyalinocytes (2.38 ± 0.08 μm, 30-35%), type II large hyalinocytes (4.41 ± 0.33 μm, 15-20%), type III small granulocytes (4.15 ± 0.26 μm, 20-25%), and type IV large granulocytes (8.26 ± 0.52 μm, 25-30%). Granulocyte types showed smaller N/C ratios than hyalinocytes. The mean haemocyte concentration was about (3.75 ± 0.65) × 107 cells ml− 1 of haemolymph. Among haemocytes, 44.7% are granular and 55.3% are agranular. These gave a relatively systematic classification scheme for haemocytes of A. irradians. Three types of granules were identified: type I, with high electron-density; type II, with low electron-density; and type III, with a middle level of electron-density, based on TEM studies. Different haemocyte types were not separated with DDGC of Percoll in this study. Both granulocytes and hyalinocytes showed a phagocytic response to the two strains of bacteria, Escherichia coli and RLOs. The phagocytic ability of granulocyte was significantly higher (41-48%) than that of hyalinocyte (9.2-11.2%). 相似文献
16.
Mohamed Alí Pereyra Oscar Vivanco‐Rojas Jos Luis Snchez‐Salgado Juan Jose Alpuche Osorno Concepcin Agundis Edgar Zenteno 《Aquaculture Research》2020,51(8):3119-3128
The crustacean haemolymph contains three main cell populations; however, it is not clear which mechanisms participate in the regulation of cells related to innate immunity. This work aimed to identify potential interleukin‐like receptors that could regulate cellular responses in Cherax quadricarinatus. By histochemical analysis with murine anti‐CD25 staining (targeting the α‐chain of the IL‐2 receptor), we identified that this antibody recognizes cytoplasmic granules in semigranular and granular haemocytes. In haemocytes stimulated with phorbol myristate acetate (PMA), increased fluorescence was observed in these cytoplasmic granules, whereas staining with a human IL‐2 antibody after stimulation with 1–10 ng/ml PMA revealed no overexpression of the receptor or oxidative burst in haemocytes. Two‐dimensional Western blot analysis of haemocyte lysates showed that anti‐CD25 identified a 27.4‐kDa protein with an isoelectric point (pI) of 7.7 and a 46‐kDa protein with a pI of 6.9. De novo sequencing of these proteins identified that they had 32% homology with a mannose‐binding lectin (MBL) from Pacifastacus leniusculus. Our results indicate that a mannose‐binding lectin‐like protein could exert a protective effect that prevents damage from other activated immune responses. 相似文献
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Gijo Ittoop Kizhakkayil C George Nandiath K Sanil Rani Mary George Kizhakkecheruvil S Sobhana & Padmini C Nisha 《Aquaculture Research》2006,37(16):1636-1643
The haemocytes of the Indian edible oyster Crassostrea madrasensis were characterized using light and electron microscopy. The light microscopic study was conducted by staining a monolayer of the haemocytes with Geimsa. Cells without granules and with a large nucleus occupying much of the cytoplasmic area were grouped as hyalinocytes. Those with lesser amounts of basophilic cytoplasmic granules were characterized as semigranulocytes and those with large amounts of a mixture of acidophilic and basophilic granules were termed as granulocytes. Ultrastructural studies also revealed the presence of three types of haemocytes. Scanning electron microscopic studies were used to study the spreading behaviour of the haemocytes. Cytochemical studies revealed the presence of acidphosphatase, peroxidase and prophenol oxidase in the cells. 相似文献
18.
流式细胞术比较研究三种对虾血细胞的分群 总被引：4，自引：1，他引：3
应用流式细胞术对墨吉对虾(Penaeus merguiensis)、南美白对虾(Penaeus vannamei)、斑节对虾(Penaeus monodon)三种对虾的血细胞类型进行了比较研究。根据其血细胞的前向角散射光(FSC)和侧向角散射光(SSC)特征的不同将血细胞分群,FSC和SSC二维图分析表明三种对虾的血细胞都可分为三个亚群:透明细胞、小颗粒细胞、大颗粒细胞。同时还对各种类型血细胞所占比例进行了比较研究,发现各个亚群血细胞所占组成比例在三种对虾之间具有相似性。其比例均为:小颗粒细胞最多,约为45%~49%;透明细胞次之,约为26%~28%;大颗粒细胞最少,约为10%~12%。 相似文献
19.
S Jayasree 《Journal of fish diseases》2009,32(4):359-365
Immune cells were identified and their interaction towards Vibrio alginolyticus, V. parahaemolyticus and V. anguillarum was studied in vitro in the penaeid shrimp, Penaeus indicus. Haemocytes were divided into agranulocytes, semi-dense granulocytes and dense granulocytes according to their morphology. Agranulocytes (100%) and 0.3–0.7% of granulocytes were actively involved in coagulation. Granulocytes were involved in in vitro phagocytosis and encapsulation of foreign materials. Phagocytosis was enhanced by prior opsonization of bacteria with cell-free shrimp haemolymph. Semi-dense granulocytes were phagocytic towards V. alginolyticus with and without opsonization at the rate of 91.1% and 83.1%, respectively ( P < 0.05 ). Granulocyte death observed after 2 h with opsonized haemolymph was 26.1%. About 64.5% of dense granulocytes and 23.2% of semi-dense granulocytes were actively involved in encapsulation, forming capsules. A spectrophotometric nitroblue tetrazolium (NBT) reduction assay was used to demonstrate the production of superoxide anions (O2 − ) by shrimp haemocytes. All the Vibrio spp. were able to induce superoxide anions (O2 − ) during phagocytosis. Live Vibrio sp. induced O2 − production in haemocytes in a dose-dependent manner. Significant activity was detected with a 40:1 bacteria to haemocyte ratio ( P < 0.05 ). NBT reduction assay for measuring the post-phagocytic killing mechanism in shrimp haemocytes might be a valuable tool for monitoring shrimp health and immunological studies. 相似文献