黄颡鱼头肾的组织发生与组织结构研究

为深入了解黄颡鱼的免疫机能,采用组织学方法,对孵化后1～48 d的黄颡鱼鱼苗头.肾的组织发生进行了观察;对5月龄幼鱼和成鱼头肾的组织结构进行了观察.结果表明,出膜后1 d,头肾仅由肾小管组成;出膜后2d,头肾小管间出现未分化的细胞团;出膜后7 d,头肾小管间出现淋巴细胞团;此后淋巴细胞及造血细胞的数量逐渐增多;出膜后26～43 d,肾小管逐渐萎缩直至完全消失,头肾完成由排泄器官向淋巴器官的转变.黄颡鱼头肾含丰富的静脉血管、血窦以及处于不同发育时期的各类免疫细胞.肾上腺组织细胞分布于头肾门静脉以及头肾组织中静脉管周边.其幼鱼头肾所含酸性颗粒细胞较成鱼头肾丰富,而成鱼头肾则具有较多的黑色素巨噬细胞,且常常在血管附近聚集形成黑色素巨噬细胞中心.     

珍珠龙胆石斑鱼血细胞发生的研究

用瑞氏染液对珍珠龙胆石斑鱼的头肾、体肾、脾脏等器官组织的涂片、印片进行染色、观察、测定和统计,以研究血细胞发生。试验结果显示,红细胞的发育经历4个阶段,分别是原红细胞、幼红细胞、红细胞和衰老红细胞,细胞体积逐渐变小;粒细胞的发育经历5个阶段,分别是原粒细胞、早幼粒细胞、中幼粒细胞、晚幼粒细胞和成熟粒细胞,中性粒细胞的体积由大变小再变大,嗜酸性粒细胞体积逐渐变小;单核细胞和淋巴细胞的发育经历3个阶段,分别是原始、幼稚和成熟,细胞体积逐渐变小。珍珠龙胆石斑鱼的各种细胞体积均较其他鱼类的细胞体积大;中性粒细胞的体积与嗜酸性粒细胞的体积差异较大。脾脏中未观察到原红细胞和原粒细胞,仅在体肾中观察到巨噬细胞和网状细胞。头肾是珍珠龙胆石斑鱼最主要的造血器官,体肾次之。头肾和体肾是珍珠龙胆石斑鱼红细胞、粒细胞的主要发生器官;头肾、体肾、脾脏是原单核细胞、原淋巴细胞发生的主要场所。     

紫红笛鲷血细胞发生研究

将体质量510~620 g的紫红笛鲷用间氨基苯甲酸乙酯甲磺酸盐麻醉后解剖,取出头肾、体肾、肝脏、脾脏,用干净的刀片居中横切、纵切,印片和涂片用Wright-Giemsa染液染色10 min,观察头肾、体肾、脾脏、肝脏4个组织印片以及外周血涂片。试验结果显示,头肾、体肾、脾脏是其造血器官。头肾能发育生成各类型血细胞,体肾能生成红细胞、淋巴细胞、粒细胞和单核细胞,脾脏能生成淋巴细胞。肝脏中未发现幼稚型血细胞。红细胞发育经过原红细胞、早幼红细胞、中幼红细胞、晚幼红细胞和红细胞5个阶段;原粒细胞发育至早幼粒细胞后,分化为嗜碱性中幼粒细胞、嗜酸性中幼粒细胞和中性中幼粒细胞,最后发育成嗜碱性粒细胞、嗜酸性粒细胞和中性粒细胞;淋巴细胞和单核细胞发育经过原始、幼稚和成熟3个阶段。血细胞在发育过程中,胞体逐渐变小,细胞核逐渐变小,染色质由疏松到致密;粒细胞中,颗粒由少到多。     

鲻脾脏和头肾的形态学与组织学研究

通过光镜技术对野生鲻(Mugil cephalus)的脾脏和头肾进行研究发现:脾脏外有薄层结缔组织纤维被膜覆盖,被膜向内延伸将脾脏分成不规则小叶,无明显小梁,含大量淋巴细胞、血细胞、粒细胞,黑色素巨噬细胞中心较多,脾脏中可见较多并行排列的动脉、静脉和神经束,有椭圆体;头肾与肾脏分离,分左右2叶。头肾为网状淋巴样组织,外被膜极薄,无明显的小梁,无肾单位,分为红血细胞聚集区和白细胞聚集区,有黑色素巨噬细胞中心,相对于脾脏,头肾中的动脉和静脉的数量较少。研究结果表明:头肾和脾脏组织是鲻的重要造血器官和免疫器官。     

急性氨氮毒性对黄颡鱼头肾巨噬细胞抗氧化及炎症相关基因表达的影响

为了研究急性氨氮胁迫对黄颡鱼头肾巨噬细胞抗氧化及炎症相关基因表达的影响,实验通过建立黄颡鱼离体头肾细胞模型,开展为期96 h的急性氨氮胁迫实验。结果显示,0.14和0.28 mg/L总氨氮处理组SOD基因的相对表达量显著低于对照组,但0.28 mg/L总氨氮处理组GPX基因的相对表达量显著高于其他组;0.14和0.28 mg/L总氨氮处理组IL-1和TNF基因的相对表达量显著低于对照组,而0.28 mg/L总氨氮处理组IL-8基因的相对表达量显著低于0.14 mg/L总氨氮处理组和对照组;吖啶橙染色检测发现,0.14 mg/L总氨氮处理组实验鱼头肾巨噬细胞内可见边缘不规则的黄绿色荧光,0.28 mg/L总氨氮处理组实验鱼头肾巨噬细胞内可见致密浓染的黄绿色荧光信号。研究表明,氨氮毒性能够影响黄颡鱼头肾巨噬细胞抗氧化相关基因的表达;细胞凋亡可能是造成鱼类氨中毒致死的主要原因之一。     

犬牙缰虾虎鱼外周血细胞的显微结构

采用Wright氏染色,对犬牙缰虾虎鱼外周血液进行观察,并进行各细胞测定,可鉴别出红细胞、血栓细胞、淋巴细胞、单核细胞、中性粒细胞和嗜碱性粒细胞,以及幼稚、正在分裂、分解和解体状态的红细胞,未发现嗜酸性粒细胞。红细胞数量多,椭圆形,具椭圆核;血栓细胞体积最小,成3~5个聚集分布,大淋巴细胞体积最大。淋巴细胞可分为大、中、小淋巴细胞。在数量上,血栓细胞最多,淋巴细胞次之,单核细胞,嗜碱性粒细胞和嗜中性粒细胞则较少。     

成年似鲇高原鳅外周血细胞及血细胞发生的观察

通过对成年似鲇高原鳅(Triplophysa siluroides)外周血细胞涂片及中肾、肝脏和脾脏组织的印片进行瑞氏(Wright’s)、过碘酸雪夫氏(PAS)和苏丹黑B(SBB)染色,观察各类血细胞形态、大小、分布及细胞化学特征。结果表明:似鲇高原鳅血细胞可分为红细胞、粒细胞、单核细胞、淋巴细胞和血栓细胞,各类血细胞的发生(除血栓细胞)均可分为原始、幼稚和成熟三个阶段。外周血中红细胞比例最多,为97.56%,白细胞中血栓细胞数量最多占52.86%,居白细胞的首位;三种粒细胞中嗜中性粒细胞数量最多占30.25%。粒细胞的主要发生场所在中肾占45.18%,红细胞的主要发生场所在脾脏,占71.62%,肝脏中未见原始血细胞。在中肾中同时可见嗜酸性、嗜碱性和嗜中性三种粒细胞的发生。各阶段的白细胞PAS染色均显示阳性,但只有各阶段的粒细胞SBB染色显示阳性。     

长鳍裸颊鲷外周血细胞的显微结构

报道了长鳍裸颊鲷(Lethrinus erythropterus)外周血细胞的显微结构。血涂片经过WRIGHT氏染色,在Olympus BX51显微镜下观察,可鉴别出红细胞、淋巴细胞、嗜中性粒细胞、嗜酸性粒细胞、嗜碱性粒细胞、血栓细胞和单核细胞等7种类型的血细胞。红细胞数量最多,多为椭圆形,具有圆核;在白细胞中,数量多少依次是血栓细胞、淋巴细胞、嗜中性粒细胞和单核细胞,嗜碱性粒细胞在外周血液中很罕见。血栓细胞体积最小,嗜中性粒细胞最大。淋巴细胞包括小淋巴细胞、中淋巴细胞和大淋巴细胞3种。     

中华鳖脾肾性水肿病病理学诊断

为探究中华鳖水肿病发病机制和发病规律,确定该病临床诊断标准,实验从若干自然发病的疑似中华鳖脾肾性水肿病的病鳖中,选取7只具有典型临床症状的病鳖,解剖并对内脏器官组织制作石蜡切片、H.E.染色后观察。结果表明:病鳖体表无病伤,不吃食,水肿症状表现不同,两只病鳖生殖器外露,全部病鳖口咽腔充血和炎症明显、黏液较少,时有口鼻出血,肝脏淤血,胃肠壁水肿和充血。病鳖脾组织髓质部红细胞数量大增和淋巴细胞聚集,并出现弥漫性坏死区域,嗜酸性粒细胞浸润,以此作出脾性水肿的病理诊断。病鳖的肾组织间质和肾小球充血,嗜酸性粒细胞浸润,远曲小管和近曲小管颗粒变性、空泡变性,肾小球坏死,近曲小管细胞坏死,以此作出肾性水肿的病理诊断。口咽腔绒毛黏膜血管扩张、充血,口腔黏膜上皮杯状细胞紧密分布,黏膜下层淋巴细胞增多,黏膜毛细血管破裂,有大量血细胞渗出,部分黏膜上皮脱落,绒毛间渗出大量坏死红细胞,嗜酸性粒细胞浸润。可以诊断口咽腔组织病理变化与该病关系密切。其他组织病变不严重。研究结果还表明中华鳖没有类似高等动物的唾液腺和腮腺,对中华鳖肿脖子病、腮(鳃)腺炎的命名提出异议,应该停止使用。     

瓦氏黄颡鱼头肾组织学观察与先天性免疫屏障研究

为探明瓦氏黄颡鱼(Pelteobagrus vachelli)头肾组织结构和先天性免疫屏障,该研究用苏木素-伊红(HE)染色法和改良James染色法对瓦氏黄颡鱼头肾的显微结构进行了观察,并使用台盼蓝活体染色法对头肾免疫屏障部位进行了定位.结果显示,瓦氏黄颡鱼头肾由被膜、淋巴细胞聚集区、粒细胞聚集区、前肾间组织、网状纤维...     

The response of coral trout (Plectropomus leopardus) to capture, handling and transport and shallow water stress

The stress response of coral trout (Plectropomus leopardus ) to wild capture or controlled shallow water stressors was investigated. Stress associated with capture from the wild, handling and transport and shallow water evoked significant changes in circulating levels of cortisol, glucose, lactate, haemoglobin (Hb) and haematocrit (Hct) in coral trout. Plasma glucose, Hb and Hct increased over 30 to 60 min in response to the controlled stressor and then returned toward unstressed levels, even if the stressor persisted. Cortisol increased to maximum levels over the 60 min following the onset of the stressor and remained elevated for 4 h in the case of a single 30 min shallow water event or 3 d in response to the capture, handling and transport. The difference between the responses of blood glucose and cortisol suggests that cortisol does not maintain blood glucose during stress in coral trout. The concentrations of circulating thrombocytes, lymphocytes and granulocytes were also significantly affected by shallow water stress. As lymphocytes and granulocytes are important immune components, reduced concentrations of these cells may explain the increases in disease susceptibility, commonly observed in stressed fish during live transport.     

Haemopoiesis in the head kidney of tilapia, <Emphasis Type="Italic">Oreochromis niloticus</Emphasis> (Teleostei: Cichlidae): a morphological (optical and ultrastructural) study

The present work focused on the histological and ultrastructural studies on haemopoiesis in the kidney of tilapia, Oreochromis niloticus. Haemopoietic tissue was found mainly in the head kidney and a small amount occurred in the mesonephros. The haemopoiesis of tilapia had the following series: erythropoiesis, granulopoiesis, thrombopoiesis, monopoiesis and lymphoplasmopoiesis. Erythropoiesis includes proerythroblasts, basophilic erythroblasts, polychromatic erythroblasts, acidophilic erythroblasts and young and mature erythrocytes. The proerythroblasts were the largest cells in the erythropoietic series. During the maturation process both the nuclear and cellular size decreased gradually due to the chromatin condensation and the progressive substitution of cytoplasmic matrix with a large amount of haemoglobin. Granulopoietic series consisted of cells with variable shape and size at different stages of maturity from myeloblasts to mature granulocytes. The promyelocytes were the largest cells in the series and were characterised by the appearance of primary (azoruphilic) granules. The maturation process involved the appearance of specific granules in the heterophilic, eosinophilic and basophilic series. It is important to mention that eosinophilic granulocytes were the dominant granulopoietic series in the haemopoietic tissue (Ht) of tilapia. Lymphopoietic series consisted of lymphoblasts, large lymphocytes, small lymphocytes and active and inactive plasma cells. Thrombopoietic series consisted of thromboblasts, prothromboblasts and thrombocytes. Thrombocytes of tilapia were nucleated and possessed a spindle shape. Melanomacrophage centres were dominant among the Ht of the head kidney. Also, monocytes were detected and shown to be large cells with an indented nucleus and cytoplasm containing numerous vesicles of different sizes and a few lysosomes.     

Morphological studies of peripheral blood cells of the Chinese sturgeon, <Emphasis Type="Italic">Acipenser sinensis</Emphasis>

The peripheral blood cells of one-year-old Chinese sturgeon (Acipenser sinensis) have been studied by light microscopy and transmission electron microscopy. The erythrocyte count was 84.86 × 10⁴ cell mm⁻³ in the peripheral blood of the fish and that of leukocytes was 2.24 × 10⁴ cell mm⁻³. The erythrocytes and four main types of leucocyte—thrombocytes, lymphocytes, granulocytes (including neutrophils and eosinophils), and monocytes, were identified in the peripheral blood. In addition to normal erythrocytes, reticulocytes and division of erythrocytes were observed. Thrombocytes were the most numerous among the leukocytes, and the number of neutrophils with lobated nuclei was larger than for other fish. The structures of the erythrocytes, lymphocytes, monocytes, granulocytes, and thrombocytes of the fish were studied. The erythrocytes were almost completely devoid of organelles, except for some mitochondria and granules. A large number of vacuoles and a few organelles were observed in cytoplasm of the monocytes. There were many microvilli on the membrane and pseudopodia-like cytoplasm bulge in the lymphocytes. The neutrophils were round or oval in shape with bilobed, trilobed, or multilobed nuclei whereas the eosinophils had big special granules, dark stained. There were many vesicles in some thrombocytes, which were related to its phagocytosis; some thrombocytes had almost no cytoplasm or organelles.     

青岛斋堂岛海域春季双桩竖杆张网网囊网目的选择性

2014年春季,采用平行作业法,以2a=16 mm(a为目脚长度)的菱形目网囊为对照网(CN),选取30 mm菱形目网囊和20 mm、30 mm、40 mm的方形目网囊(分别以30D、20S、30S和40S表示)作为实验网囊,对青岛斋堂岛附近海域的双桩竖杆张网进行了不同网目形状的网囊和不同网目尺寸方形目网囊的选择性比较试验。以多元统计分析方法进行种间选择性分析,以Logistic曲线作为选择性模型、采用极大似然估算法进行种内选择性分析。结果显示,与对照网相比,所有实验网囊均能有效释放玉筋鱼(Ammodytes personatus)、舒氏海龙(Syngnathus schlegeli)等低值渔获以及经济鱼类的幼体。比较不同网目形状网囊的渔获数据发现,方形目网囊较菱形目网囊具有更好的选择性,其中30S网囊对玉筋鱼和舒氏海龙的释放率(以尾数计)分别比30D网囊高4.61%和16.80%,对六丝钝尾虾虎鱼(Amblychaeturichthys hexanema)和日本枪乌贼(Loigo japonica)的50%选择体长(胴长)L50分别比30D网囊大38.47%和61.29%。比较不同网目尺寸方形目网囊的渔获数据发现,当实验网为20S和30S时,六丝钝尾虾虎鱼的L50分别为6.58 cm和7.20 cm,选择范围(SR)分别为3.02 cm和0.87 cm;当实验网为20S、30S和40S时,日本枪乌贼的L50分别为2.88 cm、4.03 cm和10.49 cm,SR分别为0.74 cm、0.52 cm和2.67 cm;不同尺寸的方形目网囊对这2种渔获物的L50随着网目尺寸的增大而增大。根据实验海区各种渔获物的经济价值、低值鱼类和幼鱼的释放比例以及分拣渔获物的劳动力投入等综合分析认为,30 mm方形目网囊在保护渔业资源的同时对渔业生产不会造成显著影响,建议予以推广。     

Oxidative stress in juvenile chinook salmon, Oncorhynchus tshawytscha (Walbaum)

Juvenile chinook salmon, Oncorhynchus tshawytscha (Walbaum), were held in 8–11°C freshwater, starved for 3 days and subjected to a low‐water stressor to determine the relationship between the general stress response and oxidative stress. Lipid peroxidation (LPO) levels (lipid hydroperoxides) were measured in kidney, liver and brain samples taken at the beginning of the experiment (0‐h unstressed controls) and at 6, 24 and 48 h after application of a continuous low‐water stressor. Tissue samples were also taken at 48 h from fish that had not been exposed to the stressor (48‐h unstressed controls). Exposure to the low‐water stressor affected LPO in kidney and brain tissues. In kidney, LPO decreased 6 h after imposition of the stressor; similar but less pronounced decreases also occurred in the liver and brain. At 48 h, LPO increased (in comparison with 6‐h stressed tissues) in the kidney and brain. In comparison with 48‐h unstressed controls, LPO levels were higher in the kidney and brain of stressed fish. Although preliminary, results suggest that stress can cause oxidative tissue damage in juvenile chinook salmon. Measures of oxidative stress have shown similar responses to stress in mammals; however, further research is needed to determine the extent of the stress–oxidative stress relationship and the underlying physiological mechanisms in fish.     

黄渤海夏季微藻调查

根据2011年夏季在黄渤海的采样调查分析了该海域网采浮游微藻的多样性,并从71个站位的采水样品与12个站位的拖网样品中分离了大量可培养的藻种。调查区拖网样品中共发现浮游微藻4门30属44种藻,以硅藻门(Bacillariophytas)为主,甲藻门(Dinophytas)次之。在实验室内利用毛细管法、平板法和稀释法分离纯化获得92株可培养微藻,经分子学鉴定为19种,包括9种硅藻、3种褐藻、3种不等鞭毛藻、2种绿藻、1种甲藻、1种定鞭藻。这些可培养微藻个体较小,多为微微型藻类和微型藻类,其中伪菱形藻(Pseudonitzschia sp.)和舟形藻(Navicula sp.)既能在固定样品中观察到,又能在实验室培养。圆筛藻(Coscinodiscus sp.)、梭角藻(Ceratium fusus)和夜光藻(Noctiluca scintillans)等小型藻类虽然在固定样品中所占比例较大,但是难以培养。此外,本次调查还首次在中国海域发现了Pseudobodo tremulans。黄渤海藻株的鉴定与培养不仅补充了中国微藻种质资源,还为促进微藻的研究和开发利用提供了重要材料。     

鲢小清蛋白的cDNA克隆及在大肠杆菌中的原核表达

以鲢(Hypophthalmichthys molitrix)肌肉cDNA为模板, 利用小清蛋白特异性引物进行PCR扩增, 克隆得到β小清蛋白两种不同亚型, 即Ⅰ型、Ⅱ型编码区基因。将目的基因片段连接到pET28a (+)表达载体, 并在大肠杆菌[E.coli BL21 (DE3)]中诱导表达。结果表明, 经诱导的小清蛋白重组质粒菌株有特异的蛋白表达。SDS-PAGE分析显示, 目的蛋白的分子量约为13 kD, 与预期大小一致。菌体超声破碎后发现2种亚型的小清蛋白均为可溶表达。利用Ni²⁺亲和层析柱对重组蛋白进行纯化, 得到高纯度的重组小清蛋白PVⅠ和PVⅡ。经Western Blot 鉴定, 重组小清蛋白PVⅠ和PVⅡ均能与抗鲢小清蛋白单克隆抗体反应。本研究为进一步分析小清蛋白的结构与致敏性的关系提供了重要的基础。