团头鲂外周血细胞的显微结构及细胞化学特征
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摘要
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。
The microscopic structure and cytochemical characteristics of peripheral blood cells in Megalobrama amblycephala were observed by routine Wright staining and cytochemical staining. Six types of cells were distinguished in the peripheral blood cells in Megalobrama amblycephala: erythrocytes, lymphocytes, monocytes, neutrophils, eosinophils and thrombocytes. The lymphocytes are the most abundant blood cells except erythrocytes, followed by thrombocytes, monocytes, neutrophils and eosinophils. The mature erythrocytes are mostly oval with smooth surface, oval or round nuclei whose chromatin is denser. The lymphocytes are usually round, with less cytoplasm, and the nucleus is usually eccentrically located. The monocytes are mostly round, the nucleus is round or oval, and vacuoles can be observed in the cytoplasm. Neutrophils are nearly round, and their nuclei are usually eccentrically located, lobed, kidney or oval shaped, with clear nucleoplasm boundaries. Eeosinophils are usually round, with reniform or oval nuclei, and the cytoplasm is filled with purple granules. Thrombocytes have various forms, most of which are oval, spindle, longrod shaped or teardrop shaped, and their nucleo-cytoplasmic ratio is large. The lymphocytes were positive for periodic acid-Schiff reaction(PAS), weakly positive for naphythol AS-D chloroacetate esterase(AS-DCE) and α-naphthol acetate esterase(ANAE), and negative for Sudan black B(SBB), acid phosphatase(ACP), alkaline phosphatase(AKP) and peroxidase(POX). The monocytes were strongly positive for POX and ACP, while PAS, positive for SBB, AS-DCE and ANAE were and negative for AKP. Except weakly positive for PAS and ANAE, the cytochemical staining pattern of neutrophils were the same as those of monocytes. Eosinophils showed strong positive for POX and ANAE, positive for SBB and ACP, weak positive for PAS and AS-DCE, and negative for AKP. The thrombocytes were weakly positive for PAS, AS-DCE and ANAE and negative for SBB, ACP, AKP and POX. The microstructural and cytochemical characteristics of peripheral blood cells in Megalobrama amblycephala were similar to those of other fishes, but also had obvious species specificity. The results of this investigation can be used as a basis for monitoring the health status of Megalobrama amblycephala and provide basic data for its farming and pathological diagnosis.
The microscopic structure and cytochemical characteristics of peripheral blood cells in Megalobrama amblycephala were observed by routine Wright staining and cytochemical staining. Six types of cells were distinguished in the peripheral blood cells in Megalobrama amblycephala: erythrocytes, lymphocytes, monocytes, neutrophils, eosinophils and thrombocytes. The lymphocytes are the most abundant blood cells except erythrocytes, followed by thrombocytes, monocytes, neutrophils and eosinophils. The mature erythrocytes are mostly oval with smooth surface, oval or round nuclei whose chromatin is denser. The lymphocytes are usually round, with less cytoplasm, and the nucleus is usually eccentrically located. The monocytes are mostly round, the nucleus is round or oval, and vacuoles can be observed in the cytoplasm. Neutrophils are nearly round, and their nuclei are usually eccentrically located, lobed, kidney or oval shaped, with clear nucleoplasm boundaries. Eeosinophils are usually round, with reniform or oval nuclei, and the cytoplasm is filled with purple granules. Thrombocytes have various forms, most of which are oval, spindle, longrod shaped or teardrop shaped, and their nucleo-cytoplasmic ratio is large. The lymphocytes were positive for periodic acid-Schiff reaction(PAS), weakly positive for naphythol AS-D chloroacetate esterase(AS-DCE) and α-naphthol acetate esterase(ANAE), and negative for Sudan black B(SBB), acid phosphatase(ACP), alkaline phosphatase(AKP) and peroxidase(POX). The monocytes were strongly positive for POX and ACP, while PAS, positive for SBB, AS-DCE and ANAE were and negative for AKP. Except weakly positive for PAS and ANAE, the cytochemical staining pattern of neutrophils were the same as those of monocytes. Eosinophils showed strong positive for POX and ANAE, positive for SBB and ACP, weak positive for PAS and AS-DCE, and negative for AKP. The thrombocytes were weakly positive for PAS, AS-DCE and ANAE and negative for SBB, ACP, AKP and POX. The microstructural and cytochemical characteristics of peripheral blood cells in Megalobrama amblycephala were similar to those of other fishes, but also had obvious species specificity. The results of this investigation can be used as a basis for monitoring the health status of Megalobrama amblycephala and provide basic data for its farming and pathological diagnosis.
引文
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[19]Parish N,Wrathmell A,Hart S,et al.The leucocytes of the elasmobranch Scyliorhinus canicula L.-a morphological study[J].Journal of Fish Biology,2010,28(5):545-561
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[21]Shigdar S,Harford A,Ward A C.Cytochemical characterisation of the leucocytes and thrombocytes from Murray cod(Maccullochella peelii peelii,Mitchell)[J].Fish&Shellfish Immunology,2009,26(5):731-736
[22]Jin L,Du C L,Zhang Y G.Studies on the(Ultra)microscopic structures and cytochemical characteristics of blood cells in Leiocassis longirostris Günther[J].Journal of Southwest University,2010,32(12):79-85[金丽,杜长雷,张耀光.长吻血细胞的显微、亚显微结构及细胞化学研究.西南大学学报(自然科学版),2010,32(12):79-85]
[2]Peng F,Chen X,Meng T,et al.Hematology and serum biochemistry parameters of captive Chinese alligators(Alligator sinensis)during the active and hibernating periods[J].Tissue&Cell,2018,51:8-13
[3]Zhou Y C,Xing Y N,Feng Q Y.Research advance in the haemocytes of fishes[J].Natural Science Journal of Hainan University,2003,21(2):171-176[周永灿,邢玉娜,冯全英.鱼类血细胞研究进展.海南大学学报(自然科学版),2003,21(2):171-176]
[4]Yuan S Q,Zhang Y A,Yao W J.Micro and ultrastructure of peripheral blood cells of the Mandarin fish,Siniperca chuatsi(Basilewsky)[J].Acta Hydrobiologica Sinica,1998,21(1):39-47[袁仕取,张永安.鳜鱼外周血细胞显微和亚显微结构的观察.水生生物学报,1998,21(1):39-47]
[5]Xie J H,Chen C Y.Microscopic structures of the peripheral blood cells in the pike eel Muraenesox cinereius[J].Chinese Journal of Zoology,2003,38(6):14-18[谢嘉华,陈朝阳.海鳗外周血细胞的显微结构.动物学杂志,2003,38(6):14-18]
[6]Zhang Y Z,Wang Y P,Wei Q W,et al.Composition and morphology of the peripheral blood cells of Acipenser sinensis[J].Acta Hydrobiologica Sinica,2018,42(2):322-332[张艳珍,王彦鹏,危起伟,等.中华鲟外周血细胞组成及形态观察.水生生物学报,2018,42(2):322-332]
[7]Zheng Z X,Tang Y,Fang J,et al.Ultrastructural and cytochemical properties of peripheral blood cells of Piebald Naked Carp(Gymnocypris eckloni)[J].Anatomia Histologia Embryologia,2016,46(1):17-24
[8]Da S W,Egami M I,Santos A A,et al.Cytochemical,immunocytochemical and ultrastructural observations on leukocytes and thrombocytes of fat snook(Centropomus parallelus)[J].Fish&Shellfish Immunology,2011,31(4):571-577
[9]Vázquez G R,Guerrero G A.Characterization of blood cells and hematological parameters in Cichlasoma dimerus(Teleostei,Perciformes)[J].Tissue&Cell,2007,39(3):151-160
[10]Wu W N.The formation of blood cell in Megalobrama amblycephala Yih[J].Journal of Fisheries of China,1990,14(4):328-335[吴维宁.团头鲂血细胞发生的研究.水产学报,1990,14(4):328-335]
[11]Zou S M,Li S F,Cai W Q,et al.Morphological differences of erythrocyte in autotetraploid,interploid 3n of blunt snout bream,Megalobrama amblycephala[J].Journal of Fishe-ry Sciences of China,2006,13(6):891-896[邹曙明,李思发,蔡完其,等.团头鲂同源四倍体、倍间三倍体与二倍体红细胞的形态特征比较.中国水产科学,2006,13(6):891-896]
[12]Xu W R.Clinical Hematology and Blood test Experimental Guidance[M].Beijing:People’s Health Press.2005,8,6-12[许文荣.临床血液学和血液检验实验指导.北京:人民卫生出版社.2005,8,6-12]
[13]Tavares-Dias M.A morphological and cytochemical study of erythrocytes,thrombocytes and leukocytes in four freshwater teleosts[J].Journal of Fish Biology,2006,68(6):1821-1833
[14]Gong X L,Wang M X,Wang D H,et al.The morphological characteristics of the peripheral hematocytes in bullfrog(Rana catesbeiana)[J].Chinese Journal of Zoology,2015,50(4):591-599[龚小玲,王米雪,汪德海,等.牛蛙外周血细胞的形态学特点.动物学杂志,2015,50(4):591-599]
[15]Guo X G,Zhang Y G,Wang Z J,et al.Study on blood cell of Bufo gargarizans[J].Sichuan Journal of Zoology,2002,21(4):211-214[郭宪光,张耀光,王志坚,等.中华蟾蜍血细胞的观察.四川动物,2002,21(4):211-214]
[16]Cao W Z,Liu Y,Liang B,et al.Development of haemocytes in peripheral blood and hematopoietic organs of zebra fish,Danio rerio[J].Fisheries Science,2014,33(7):403-409[曹文芝,刘云,梁冰,等.斑马鱼外周血液及造血器官血细胞发生的观察.水产科学,2014,33(7):403-409]
[17]Dai Y G,Yue X T,Yin B Y.Characteristics of peripheral hematocytes in endangered fish Onychostoma rara[J].Chinese Journal of Zoology,2014,49(6):875-885[代应贵,岳晓烔,尹邦一.濒危鱼类稀有白甲鱼外周血细胞特征.动物学杂志,2014,49(6):875-885]
[18]Jin L,Zhao N,Huang L,et al.Structure and cytochemical characteristics of blood cells in Myxocyprinus asiaticus[J].Acta Hydrobiologica Sinica,2011,35(3):550 -556[金丽,赵娜,黄林,等.胭脂鱼外周血细胞的显微、超微结构与细胞化学观察.水生生物学报,2011,35(3):550-556]
[19]Parish N,Wrathmell A,Hart S,et al.The leucocytes of the elasmobranch Scyliorhinus canicula L.-a morphological study[J].Journal of Fish Biology,2010,28(5):545-561
[20]Zhao H P,Zhao H T,Jin L,et al.Microscopic structures of peripheral hematocytes in Sinilabeo rendahli[J].Fisheries Science,2005,24(6):24-27[赵海鹏,赵海涛,金丽,等.华鲮外周血细胞显微结构观察.水产科学,2005,24(6):24-27]
[21]Shigdar S,Harford A,Ward A C.Cytochemical characterisation of the leucocytes and thrombocytes from Murray cod(Maccullochella peelii peelii,Mitchell)[J].Fish&Shellfish Immunology,2009,26(5):731-736
[22]Jin L,Du C L,Zhang Y G.Studies on the(Ultra)microscopic structures and cytochemical characteristics of blood cells in Leiocassis longirostris Günther[J].Journal of Southwest University,2010,32(12):79-85[金丽,杜长雷,张耀光.长吻血细胞的显微、亚显微结构及细胞化学研究.西南大学学报(自然科学版),2010,32(12):79-85]