长吻(鱼危)血细胞发生的研究

用Wright-Giemsa和PAS染色对长吻(鱼危)头肾、肾脏、脾脏、肝脏等器官组织的涂片、印片染色观察发现,头肾、肾脏和脾脏是其主要造血器官.红细胞、粒细胞和淋巴细胞主要在肾脏和头肾中发生,其次是脾脏.单核细胞则主要在肾脏和脾脏中发生,头肾中也有少量单核细胞产生.肝脏中无原始型血细胞,可能不是其造血器官.红细胞的发育经历四个阶段,其胞体体积经历了由大到小,由小到大再变小的"两大两小"发育过程;粒细胞的发育经历五个阶段,其胞体体积均由大变小,双叶或多叶核的粒细胞可能是衰老的粒细胞亦即核的分叶是粒细胞衰老的标志;淋巴细胞和单核细胞的发育各经历了三个阶段,两者发育成熟过程中胞体体积均由大变小.巨噬细胞由单核细胞发育而来.原血细胞和部分早期幼稚血细胞可以进行有丝分裂,部分成熟红细胞和血栓细胞可以进行直接分裂.红细胞在整个发育过程中,PAS反应均呈阴性,各类白细胞的发育过程中,PAS反应由阴性到阳性并逐渐增强,这显示随着白细胞的逐渐发育成熟,细胞内糖原物质含量逐渐增多.     

齐口裂腹鱼血细胞的发生

为了探讨齐口裂腹鱼(Schizothorax prenanti)血细胞发生发育的模式,采用Wright’s、碘酸雪夫氏(PAS)和苏丹黑B(SBB)染色方法对齐口裂腹鱼的头肾、中肾、脾和肝组织印片进行染色,了解其发生的具体组织和发育的一般过程。结果表明:齐口裂腹鱼血细胞可分为红细胞系、淋巴细胞系、单核细胞系、粒细胞系和其他细胞,且这些细胞系的发育均经过3个阶段,即原始阶段、幼稚阶段、成熟阶段。通过观察和统计不同阶段各种血细胞的形态、大小、比例及染色特征,发现原始阶段的血细胞体积较大,其中原始单核细胞最大,大小为(18.90±1.59)μm×(16.32±0.70)μm,在4种组织中原始阶段的红细胞和单核细胞在头肾中所占比例最大,分别为0.64%和0.59%,原粒细胞和原淋巴细胞分别在中肾和脾中比例最大,在肝中少量存在,此外在各组织印片中均发现血栓细胞的存在,在肝中发现巨噬细胞。因此头肾、中肾和脾是齐口裂腹鱼的主要造血组织,少量细胞在肝中产生。粒细胞发育过程中,除原始粒细胞PAS染色阴性外,其他阶段均呈阳性,且阳性随不断成熟逐渐增强;单核细胞从原单核细胞到成熟的单核细胞,PAS阳性逐渐增强;而SBB染色发现,粒细胞、淋巴细胞及单核细胞均呈阳性,未成熟的细胞染色程度不一致,成熟的细胞阳性染色最为强烈。在红细胞系发育过程中经历了细胞由大变小再变大的过程,而粒细胞和淋巴细胞发育过程仅出现由大变小的过程,在红细胞的发育过程中还出现了染色质固缩和血红蛋白增加。     

卵形鲳鲹血细胞发生的观察

对卵形鲳鲹(Trachinotus ovatus)肝脏、脾脏、头肾和体肾4种组织器官内各种血细胞的发生和外周血液的血细胞分类组成进行了观察。结果表明:卵形鲳鲹的红细胞主要在脾脏、体肾和头肾中发生;淋巴细胞主要在体肾、头肾和脾脏中发生;粒细胞主要在头肾和体肾中发生;单核细胞在肝脏、头肾和脾脏中均有发生;而血栓细胞的发生在这4种组织中均未观察到。卵形鲳鲹在外周血液中的白细胞包括淋巴细胞、血栓细胞、嗜中性粒细胞和单核细胞。在这几种白细胞中,单核细胞数量最少,很难观察到,数量少于1%;嗜中性粒细胞稍多,有(9.25±5.64)%;血栓细胞占(38.23±21.86)%;淋巴细胞最多,达(52.52±21.65)%。     

团头鲂外周血细胞的显微结构及细胞化学特征

采用常规瑞氏染色和细胞化学染色方法对团头鲂(Megalobrama amblycephala)外周血细胞的显微结构及细胞化学特征进行了观察。在团头鲂外周血细胞中可区分出六类细胞: 红细胞、淋巴细胞、单核细胞、嗜中性粒细胞、嗜酸性粒细胞和血栓细胞。其中淋巴细胞是除红细胞外含量最多的细胞, 其次分别为血栓细胞、单核细胞、嗜中性粒细胞、嗜酸性粒细胞。成熟红细胞多为卵圆形, 表面光滑, 胞核呈椭圆形或圆形, 染色质较为致密; 淋巴细胞多呈圆形, 胞质较少, 胞核常偏位; 单核细胞多为圆形, 胞核呈圆形或椭圆形, 胞质内可见空泡状结构; 嗜中性粒细胞近似圆形, 胞核常偏于细胞一侧, 呈分叶状、肾形或椭圆形, 核质界限清晰; 嗜酸性粒细胞一般为圆形, 胞核为肾形或椭圆形, 胞质中充满紫红色颗粒; 血栓细胞形态多样, 主要有椭圆形、纺锤形、长杆状和泪滴形, 核质比较大。淋巴细胞呈α-醋酸萘酚酯酶(ANAE)阳性, 呈过碘酸-雪夫(PAS)、氯乙酸AS-D萘酚酯酶(AS-DCE)弱阳性, 呈苏丹黑B(SBB)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)及过氧化物酶(POX)阴性; 单核细胞呈POX、ACP强阳性, PAS、SBB、AS-DCE和ANAE为阳性, 呈AKP阴性; 嗜中性粒细胞除PAS和ANAE为弱阳性外, 其他染色结果和单核细胞相同; 嗜酸性粒细胞呈POX、ANAE强阳性, SBB、ACP阳性, PAS及AS-DCE则为弱阳性, 呈AKP阴性; 血栓细胞呈PAS、AS-DCE及ANAE弱阳性, 呈SBB、ACP、AKP及POX阴性。团头鲂外周血细胞的显微结构及细胞化学特征与其他鱼类具有相似之处, 但亦有其明显的物种特异性。该研究结果可作为监测团头鲂健康状态的依据, 为其养殖及病理诊断提供基础资料。     

扬子鳄外周血细胞的超微结构

对扬子鳄血细胞亚显微结构进行透射电镜观察。结果表明：在外周血细胞中可区分出红细胞、嗜中性粒细胞、嗜酸性粒细胞、嗜碱性粒细胞、单核细胞、淋巴细胞以及血栓细胞。电镜图像显示：与其它爬行动物相比,各类血细胞体积较小并具有不同程度的变形运动及吞噬能力。红细胞的特征是长梭形,其长短径之比值超过3.0。在外周血液中发现了多种未成熟型血细胞;进入外周血液中的红细胞普遍已分化到一定程度,但有的仍须在外周血液中完成其最后的发育;单核细胞在外周血液中经历了从幼单核细胞到成熟单核细胞的过程,并描述了从未成熟到成熟阶段细胞的超微结构的变化;嗜中性粒细胞在外周血液中经历了从早幼粒细胞、中幼粒细胞,晚幼粒细胞到成熟粒细胞的过程,描述了嗜中性粒细胞各个发育阶段细胞的超微结构及特征,并讨论了嗜天青粒细胞和嗜中性粒细胞的关系。     

养殖齐口裂腹鱼外周血细胞显微观察

养殖齐口裂腹鱼Schizothorax prenanti外周血经瑞氏染色液染色,可鉴定出红细胞、嗜中性粒细胞、血栓细胞、淋巴细胞、单核细胞,一些未成熟的红细胞和少量进行无丝分裂的红细胞,未观察到嗜碱性粒细胞和嗜酸性粒细胞.在外周血中还可观察到3个阶段的嗜中性粒细胞:中性晚幼粒细胞胞体为圆形或近圆形,胞浆呈淡粉红色,量丰富,其中含有较多红色的细小颗粒,胞核呈肾形;中性杆状核粒细胞胞核较中性晚幼粒细胞的凹陷更强,呈"S"、"C"等形状;中性分叶核粒细胞占多数,胞体近圆形,核至少分成两叶,多达五或六叶.血栓细胞呈圆形、近圆形、梨形、纺锤形、杆状等多种形态,一个或多个聚集在一起.外周血细胞中,血栓细胞体积最小、数量最多,单核细胞体积最大、数量最少.     

军曹鱼血液指标及血细胞发生的观察

测定军曹鱼的血液指标,红细胞密度为2．69±0．86×106个/mm3,白细胞密度为1．50±0．09×104个/mm3;血 红蛋白含量为7．42±0．22g/L,红细胞渗透脆性为0．43±0．07g％,红细胞沉降速率为1．18±0．46mm/h。观察军曹鱼 外周血液涂片,可区分出红细胞、血栓细胞、淋巴细胞、嗜中性粒细胞和单核细胞,但没有发现嗜酸性粒细胞和嗜碱 性粒细胞。在外周血液涂片观察中还发现了较多未成熟的红细胞和嗜中性粒细胞以及少量正在分裂的红细胞。 血栓细胞、淋巴细胞、嗜中性粒细胞和单核细胞在白细胞中所占比例分别为61．20±6．30％,16．60±3．28％,16．00± 3．61％和6．20±3．90％。对肝脏、脾脏、头肾和中肾等四种造血组织进行了涂片观察,军曹鱼的血细胞主要在头肾 和肾脏产生;脾脏是军曹鱼粒细胞发生的另一个场所,而肝脏也具有产生粒细胞和淋巴细胞的作用。       

草鱼肾脏和脾脏血细胞发育过程的观察

鱼类肾脏、脾脏是机体造血的主要器官。印迹涂片显示:草鱼肾脏和脾脏内血细胞的发育过程大致经历了三个阶段,即原始阶段、幼稚阶段、成熟阶段。本文着重描述了各阶段细胞的形态特征井对草鱼血细胞的发育及命名等问题作了初步的探讨。       

鳜鱼外周血细胞显微和亚显微结构的观察

本文报道了鳜鱼外周血细胞的显微和亚显微结构。血涂片经过染色,可鉴别出红细胞、血栓细胞、淋巴细胞、单核细胞和嗜中性粒细胞;还见到幼稚的正在分裂的红细胞,提示红细胞亦可在外周血液中通过直接分裂产生。白细胞中,血栓细胞体积最小,嗜中性粒细胞体积最大;单核细胞数目最少,血栓细胞数目最多。电镜下,红细胞中可见线粒体和高尔基复合体;淋巴细胞线粒体中可见类似髓样体的板层状结构;血栓细胞和单核细胞与其它鱼类的基本     

牛蛙外周血细胞的形态学特点

本研究对雌雄牛蛙(Rana catesbeiana)外周血细胞的组成、形态、大小和数量进行了观察和统计。牛蛙外周血细胞由红细胞、白细胞以及血栓细胞组成,其中红细胞体积最大,平均大小(长径×短径)为(25.68±1.88)μm×(16.49±1.53)μm,扫描电镜下发现红细胞表面光滑;血栓细胞呈卵圆形或纺锤形,其体积最小,平均大小为(8.62±1.04)μm×(7.47±1.11)μm;白细胞由淋巴细胞、单核细胞、浆细胞、嗜中性粒细胞、嗜酸性粒细胞和嗜碱性粒细胞组成,扫描电镜下白细胞表面粗糙不平,有许多不规则的凸起。白细胞中淋巴细胞最多,其中小淋巴细胞约占白细胞的32.66%±4.29%,大淋巴细胞约占6.03%±1.54%;嗜碱性粒细胞最少,只占4.78%±0.83%;浆细胞胞体大小不一,常呈椭圆形,平均大小为(23.51±0.59)μm×(22.86±0.67)μm;此外,牛蛙外周血细胞中单核细胞、淋巴细胞和嗜碱性粒细胞的数量比例以及淋巴细胞和嗜碱性粒细胞的大小均有性别的差异(P0.05)。     

Composition of main haematopoietic compartments in normal and bled channel catfish

Differential cell counts showed that the head and trunk kidney of control and bled channel catfish Ictalurus punctatus had myeloid characteristics. They contained lymphoid and granuloid cells, thrombocytes, erythroid and agranular cells in decreasing order of abundance (%). Among the blast and precursor cells, the most numerous erythroid ones were followed by granuloid, lymphoid and agranular ones. The main changes after blood withdrawal were the decrease of thrombocytes and the increase of precursor cells in both kidney parts. In the group examined 7 days after bleeding the head kidney had a higher percentage of erythroid cells and lymphocyte precursors than the trunk kidney while the latter had more granuloid cells and their precursors. Basophils were present ( c . 1%) in both regions of the kidney of all groups. The spleen was predominantly a lymphatic organ. It contained c . 80% lymphoid cells, a higher incidence of granulated lymphocytes than in kidneys, 15% thrombocytes and 1.4% agranular cells. Blood withdrawal caused an increase of thrombocytes, a decrease of lymphoid cells and an increase of erythroid precursors in the spleen. The last probably stemmed from the circulation. While haematocrit values failed to indicate the anaemic state in the bled groups, the differential red blood cell count showed dramatic differences between the control and bled groups as well as between the two groups in different stages of recuperation from the blood loss.     

Morphogenesis of blood cell lineages in channel catfish

The morphogenesis of blood cell lineages in channel catfish Ictalurus punctatus , from head and trunk kidney and spleen imprints as well as from blood smears of bled and control fish, showed that early maturation stages resembled those in higher vertebrates. The erythroid lineage consisted of the proerythroblast, erythroblasts (basophilic, polychromatic, orthochromic), young erythrocyte and erythrocyte. The rare bilobed erythrocyte seemed to be a cell in apoptosis while old erythrocytes and erythroplastids represented remnants of this process. Maturation stages of neutrophils and basophils encompassed the granuloblast, young progranulocyte, progranulocyte and metagranulocyte. The basophilic lineage was regularly present in kidneys, rare in spleen and absent from blood. It contained large Sudan Black and PAS-negative, water soluble granules and small PAS-positive ones. Lymphocytes with azurophilic granulation occured regularly in kidneys and spleen. Monoblasts and promonocytes in kidneys preceded monocytes. A phagocytic lineage devouring apoptotic blood cell remnants was present in kidneys and spleen. Its youngest identified stage (promacrophage) resembled more a granuloid cell without granules than a monocytoid one. The larger, young macrophages contained a few to several ingestions and the very large mature macrophages were loaded with them. The latter two stages corresponded to cells in melano-macrophage centres (macrophage aggregates). Precursor stages of the thrombocyte were not identified.     

长薄鳅外周血细胞的显微结构和细胞化学特征研究

长薄鳅外周血细胞可分为红细胞、中性粒细胞、单核细胞、淋巴细胞和血栓细胞.在数量上,中性粒细胞、单核细胞、淋巴细胞和血栓细胞占白细胞总数的百分比分别是17.06%、5.83%、28.16%和48.94%.细胞化学染色显示所有白细胞均含有糖原物质,所有红细胞均不含酸性磷酸酶,中性粒细胞、单核细胞、淋巴细胞和血栓细胞均含有酸性磷酸酶.非特异件酯酶染色显示单核细胞呈阳性反应,中性粒细胞、淋巴细胞和血栓细胞均为部分呈阳性反应.所有细胞的碱性磷酸酶、过氧化物酶、苏丹黑显色反应均呈阴性.     

Cell types in peripheral blood of the nurse shark: An approach to structure and function

Ultrastructural and functional studies were carried out on nurse shark (Ginglymostoma cirratum) peripheral blood cells in order to identify cells of definitive morphology and specific function. Along with erythrocytes and thrombocytes, four morphologically distinct leucocytes are recognized in peripheral blood: two types of granulocytes, the ‘eosiniphil’ and the ‘granulocyte’, and two mononuclear agranulocytic cells, one resembling mammalian macrophage and monocyte, the other resembling mammalian lymphocyte. Also present in peripheral circulation are blast-like cells and mitotic cells. In vitro phagocytosis was demonstrated by the monocyte-macrophage and the granulocyte while thrombocytes, eosinophils and lymphocytes showed no phagocytic activity in the system studied. It is stressed that care must be used in drawing functional analogies between blood cells of a mammal and an elasmobranch on the basis of morphological similarity alone.     

A monoclonal antibody recognising a surface marker on rainbow trout (Oncorhynchus mykiss) monocytes

The characterisation of a monoclonal antibody (mab 45) reacting with phagocytic leucocytes isolated from blood and spleen of rainbow trout (Oncorhynchus mykiss, Walbaum) is described. The surface marker labelled by this mab is expressed at relative low levels on the membrane of large, nearly nongranulated trout leucocytes, and having the typical morphology of monocytes in flow cytometry (Kfoury et al., 1999, Fish Pathology, 34, 1-6). No reaction of mab 45 with granulocytes, lymphocytes or thrombocytes was detected. In spleen and head kidney, large, polymorphonuclear leucocytes were immunostained. The mab most strongly recognised an antigen of 48 kDa prepared from trout leucocytes of different organs, but not in trout plasma. In an in vitro phagocytosis assay trout monocytes were stained with mab 45 after phagocytosis of Aeromonas salmonicida labelled with the lipophilic fluorescent cell surface linker PKH26. However, previous binding of mab 45 on trout leucocytes did not inhibit the phagocytosis of A. salmonicida particles. Using mab 45, the dynamics of monocytes in blood, spleen and peritoneal cavity could be demonstrated after intraperitoneal injection of trout with inactivated A. salmonicida. The described mab serves as a useful tool to investigate the involvement of monocytes/macrophages in immune reactions of trout to a variety of pathogens.     

Preliminary observation of blood cells and hematopoietic organs in Lutjanus herenbergi and Lutjanus flaviflammus

The morphology of differentiated and differentiating cells of the red and white series in Lutjanus herenbergi and in Lutjanus flaviflammus is described. Early stages of red and white blood cells may be found only in smears of hemopoietic organs. Polychromatic erythroblasts, myelocytes and lymphoblasts may also occasionally be found in blood smears. Mature blood cells may be found both in blood smears and in hemopoietic organs. Differential white cell counts seem to demonstrate that the granulocytic series elements are the most common leukocytes in blood smears. Almost all granulocytes may be classified in the first three Arneth classes. An analysis of hemopoietic organs in these species was also performed. It was found that the only organs carrying on a hemopoietic function are the kidney and the spleen. The kidney is essentially a site of granulocytic differentiation while the spleen is a lymphopoietic organ. An erythropoietic activity may generally be observed in the kidney although weak erythropoietic activity may at times be found in the spleen.     

Ultrastructure of the peripheral blood leucocytes of Oreochromis mossambicus

Peripheral blood leucocytes were examined for their fine structure and their in vivo response to colloidal carbon particles and bacteria, Aeromonas salmonicida . Four main types of leucocyte were identified: lymphocytes, thrombocytes, monocytes and granulocytes. Previously, two types of granulocyte had been identified but at the electron microscope level a third type could be distinguished in the peripheral blood. Monocytes and the type 1 and type 3 granulocytes were found to be phagocytic.     

Measuring some flounder (Platichthys flesus L.) innate immune responses to be incorporated in effect biomonitoring concepts

For an implementation of innate immune responses of flounder (Platichthys flesus) in an integrated biological effect monitoring concept, leucocytes were isolated from peripheral blood, head kidney and spleen, and analysed for their capacity to mount a respiratory burst response upon phorbol ester stimulation. Responding cells were identified by reduced nitro-blue-tetrazolium salt deposits and by dihydro-rhodamine fluorescence in light microscope and flow cytometric analysis. Responding cells were found in head kidney derived cell suspensions rather than in peripheral blood or spleen. Parallel cytometric and microscopic analysis indicated that responding cells had a granulocyte or monocyte morphology, were alpha-naphtyl-esterase or myeloperoxidase positive and in flow cytometry exhibited a characteristic forward and side scatter (FSC/SSC) pattern. These cells represented 30–40% of head kidney derived cell suspensions and only 4–5 % of peripheral blood and spleen. In order to reduce sampling effort in field studies, leucocyte cell suspensions derived from flounder head kidney could be used in respiratory burst assays without further enrichment protocols. This paper combines, for the first time, conventional and cytometric analysis of phagocytes derived from flounder peripheral blood and head kidney. Communicated by H. v. Westernhagen, A. Diamant     

A cytochemical and light microscopical study of the peripheral blood leucocytes of Oreochromis mossambicus, Cichlidae

The morphological characteristics of the peripheral leucocytes of the cichlid O. mossambicus were studied using light microscopy. Cytochemical procedures and phagocytic studies were employed. Four main types of leucocytes could be recognized: lymphocytes, thrombocytes, monocytes and granulocytes. Two distinct types of granulocyte were identified. Monocytes and one type of granulocyte were found to be phagocytic.