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1.
殷国俊 《水产学报》2004,28(6):628-632
用密度梯度离心分离鲤头肾中的巨噬细胞和嗜中性粒细胞。用RPMI细胞培养液对分离的细胞进行原代培养,在细胞培养液中加入不同浓度的黄芪水提取液来研究黄芪对鲤头肾中免疫细胞非特异性免疫应答的影响。黄芪水提取液单独使用时能刺激鲤头肾中巨噬细胞的增殖,但不能刺激巨噬细胞的氮暴发活性。用黄芪和脂多糖(LPS)混合刺激巨噬细胞和中性粒细胞时,黄芪水提取液能显著提高脂多糖刺激所产生的一氧化氮的产量。研究结果表明,黄芪不仅能刺激巨噬细胞数量的增加,而且能协同脂多糖增强头肾中免疫细胞的功能,从而对机体的非特异性免疫起调节作用。  相似文献   

2.
借鉴哺乳动物内皮细胞体外培养技术,结合鱼类细胞自身特点,纯化培养鲤血管内皮细胞并传至3代,以此作为粘附材料。应用淋巴细胞分离液离心技术并结合玻璃粘附法分离纯化鲫、草鱼外周血中淋巴细胞、单核细胞,并与上述3代内皮细胞进行免疫粘附试验,同时进行免疫粘附动力学观察。结果显示,鲫、草鱼两类细胞对内皮细胞的粘附率分别为0.09±0.013,0.20±0.018;0.11±0.015,0.21±0.023。表明鲤科鱼类不同种的同类细胞粘附率差异不大,而同种不同类细胞则差异显著。动力学观察分析表明,淋巴细胞粘附较快,60min进入平台期;单核细胞粘附慢,120min进入平台期。初步证明鱼类内皮细胞具有免疫介导作用。  相似文献   

3.
对40尾野生青石爬鮡进行了血液生化指标、血细胞分类与发生研究。测得血浆的总蛋白为(48.35±3.60) g/L,白蛋白为(8.73±0.54) g/L,球蛋白为(39.63±3.10) g/L,血浆白蛋白与球蛋白比为(0.22±0.01),甘油三酯为(5.05±2.14) mmol/L,总胆固醇为(16.66±3.64) mmol/L,极低密度脂蛋白为(1.01±0.43) mmol/L,葡萄糖为(5.27±2.43) mmol/L。血液生化指标检测结果表明,青石爬鮡血浆中总蛋白与球蛋白、甘油三酯与胆固醇较高,葡萄糖较低,具有活动量较大和冷水性鱼类的特点,主要利用脂肪和蛋白质作为能量。细胞学显示出其外周血液包含红细胞,核影,淋巴细胞,血栓细胞,嗜中性粒细胞和单核细胞。血涂片计数为红细胞96.58%,其中成熟红细胞占65.92%,幼稚红细胞占30.66%。白细胞为3.42%,其中嗜中性粒细胞0.14%、单核细胞0.07%、淋巴细胞2.37%、血栓细胞0.85%。白细胞中淋巴细胞和血栓细胞数量较多。原始血细胞主要在中肾和头肾中,在脾脏中也有部分原始白细胞,而肝脏和外周血液中未见原始血细胞。中肾和头肾是青石爬鮡的主要造血器官,其中红细胞主要由头肾产生,白细胞主要由中肾产生。脾脏亦产生少量白细胞。  相似文献   

4.
草鱼肠巨噬细胞的分离培养与鉴定   总被引:1,自引:0,他引:1  
陶会竹  肖宁  赵雨婷  房慧  李槿年 《水产学报》2018,42(10):1606-1614
为了建立草鱼肠巨噬细胞的分离培养与鉴定方法,本研究在采用刮除法结合胶原酶IV消化法制备肠黏膜固有层单细胞悬液的基础上,使用鱼类脏器单核细胞分离试剂盒分离肠巨噬细胞,再经差异贴壁法纯化肠巨噬细胞,最后,用含10%胎牛血清和5%草鱼血清的RPMI 1640完全培养液,在28°C、5%CO2条件下原代培养肠巨噬细胞,并通过细胞形态学检查、分子标记检测和功能验证实验加以鉴定。结果显示,每尾鱼(约250 g)肠巨噬细胞的获得量约为3×107个,细胞活率为99.6%,纯度达到95%以上;倒置显微镜观察发现,原代培养4~5 d的贴壁细胞多呈圆形或多边形,体积明显增大,细胞汇合度达到95%;光镜和电镜观察到染色后的贴壁细胞具有巨噬细胞的形态结构特征,贴壁细胞经荧光定量PCR在m RNA水平检测到巨噬细胞特异性标志分子(草鱼巨噬细胞集落刺激因子受体),功能实验证实贴壁细胞具有吞噬功能,脂多糖能够显著提高其呼吸暴发活性。本研究首次成功建立了草鱼肠巨噬细胞的分离培养与鉴定方法,为开展口服疫苗诱导的草鱼肠黏膜免疫应答研究提供细胞模型。  相似文献   

5.
鱼类树突状细胞研究进展   总被引:1,自引:1,他引:0  
树突状细胞(DCs)是目前已知的体内功能最强的抗原递呈细胞,是唯一能够激活初始T淋巴细胞反应的细胞,在先天性免疫、适应性免疫以及维持自身免疫耐受方面具有重要的作用,因此一直是免疫学研究的重要领域。本文简要综述了DCs的类型及其在动物体内的功能、各类DCs的细胞标记。总结了鱼类DCs的分离、纯化方法和形态学观察方法;现有研究表明,鱼类DCs具有吞噬细菌、刺激T细胞增殖、诱导CD4+T细胞的活化、表达DCs的标记基因、被Toll样受体的配体激活、迁移能力、引起混合淋巴细胞反应等生物学功能;不同鱼类DCs的分子标记并不完全一样;鱼类的头肾、肾、鳃、皮肤、胸腺、脾、肠等均有DCs的分布。目前,对鱼类DCs的研究虽然取得了一定进展,但仍有许多重要问题需要解决:①鱼类DCs目前缺乏明确的细胞标记,加强这方面的研究有助于提高鱼类DCs的分离、体内分布与功能的研究水平;②加强和完善鱼类DCs的分离、培养技术的研究,掌握各种鱼类DCs的分离培养方法;③加强鱼类DCs在抗原递呈中的功能研究,对深入分析鱼类免疫机理,合理设计和应用疫苗,具有重要的理论指导意义。  相似文献   

6.
对低等脊椎动物血细胞分类、发生和功能的研究进展进行了综述.低等脊椎动物血细胞分红细胞和白细胞2类.其中白细胞又可分为淋巴细胞、血栓细胞、单核细胞、嗜中性粒细胞、嗜酸性粒细胞、嗜碱性粒细胞和浆细胞等.有关低等脊椎动物血细胞发生的研究报道较少,并主要集中在鱼类.鱼类血细胞发生的部位包括胸腺、头肾、体肾、脾脏、肝脏和肠粘膜下层等.血细胞的功能与维持动物机体内环境的稳定性有密切关系,低等脊椎动物的血细胞还具有吞噬、分泌等功能,并参与了机体伤口的修复和免疫防御等生理过程.  相似文献   

7.
为了更加了解草鱼B淋巴细胞吞噬活性,本研究采用实时荧光定量PCR(qRTPCR)检测了IgM在胚胎发育中的表达量,并且检测了IgM在不同组织中的分布以及细菌刺激下IgM的转录情况。结果显示,IgM在卵裂期到出膜前期的表达量变化不明显,在出膜后开始显著增加;IgM在检测的组织中均有分布,在头肾中表达量最高,并且其表达量在不同细菌的刺激下均显著上调。从草鱼外周血中分离纯化得到B淋巴细胞,并通过吉姆萨染色、qRT-PCR和间接免疫荧光进行了验证。细菌吞噬实验结果显示,B淋巴细胞对嗜水气单胞菌具有一定的吞噬能力,并随孵育时间增加而逐渐增强。细菌刺激B淋巴细胞后活性氧(reactive oxygen species, ROS)和一氧化氮(nitric oxide,NO)释放量显著上升;血清调理实验结果显示,通过血清调理可以显著促进B淋巴细胞ROS的释放,然而对NO的释放水平没有显著影响。研究表明,草鱼B淋巴细胞对细菌具有一定的吞噬能力,并且可以通过呼吸爆发等非特异性免疫的方式直接参与抗菌免疫。  相似文献   

8.
IL-8在LPS诱导的草鱼炎症过程中的表达特征   总被引:1,自引:0,他引:1  
采用原核表达获得草鱼白细胞介素-8(IL-8)重组蛋白,并以腹腔与皮下组织交替注射的方法免疫小鼠,制备抗草鱼IL-8的多克隆抗体。采用免疫印迹(Western blot)和酶联免疫吸附(ELISA)技术检测抗体的特异性与效价,运用流式细胞术分析健康草鱼及细菌脂多糖(LPS)刺激后草鱼各免疫相关组织中IL-8的表达特征。结果显示,草鱼IL-8多克隆抗体效价可达1∶40 000。健康草鱼的胸腺、头肾、肝脏、肠道、鳃、脾脏、体肾等组织均表达IL-8蛋白,其中,头肾和鳃中表达量较高。LPS刺激后,各组织IL-8蛋白表达量均显著升高,其中,肠道、肝脏、肾脏在LPS刺激4 h后IL-8蛋白上调达最高峰,头肾、脾脏、鳃于LPS刺激后12 h时表达量最高,胸腺则在24 h时达最高峰。研究表明,IL-8参与了草鱼炎症应答;IL-8表达量的升高,可作为早期炎症监测指标,应用于养殖鱼类炎症性疾病的预警指标。  相似文献   

9.
鱼类免疫应答机制研究进展   总被引:1,自引:0,他引:1  
鱼类免疫应答可以分为固有免疫和适应性免疫,但固有免疫发挥主要作用。固有免疫对病原体的识别是通过模式识别受体PRR与病原相关分子模式PAMP的相互结合实现,这与哺乳类相似。但为适应水生生活,鱼类固有免疫对PAMP的识别范围更广,免疫应答的启动条件更低。固有免疫的效应细胞主要是单核/巨噬细胞、嗜中性粒细胞、自然杀伤细胞等,具有吞噬和杀伤功能,还可分泌多种免疫相关的细胞因子,介导发生炎症反应。适应性免疫中,T淋巴细胞通过抗原提呈细胞分解吸收抗原,由主要组织相容性复合物(MHC)类分子递送到细胞表面才能识别。B淋巴细胞分泌产生以免疫球蛋白Ig M为主的抗体分子,而发挥抗体中和作用及免疫调理作用的Ig G在鱼类中比较少见,说明鱼类抗体的免疫功能还处于较低水平。本文综述了近二十年内鱼类免疫应答机制的相关研究进展,为进一步了解鱼类免疫应答机制提供参考。  相似文献   

10.
壳聚糖通过甲状腺激素对草鱼免疫功能的调节   总被引:3,自引:0,他引:3       下载免费PDF全文
以初始体质量为(1.04±0.01)g的草鱼(Ctenopharyngodon idella)为对象,在探讨内分泌激素—甲状腺激素对草鱼免疫功能影响的基础上,研究壳聚糖对草鱼免疫功能的内分泌调控。实验分别制作含甲状腺激素饲料(在基础饲料中分别添加0mg/kg、1mg/kg、5mg/kg、10mg/kg饲料的甲状腺激素)和含壳聚糖饲料(在基础饲料中分别添加0、0.25%、0.50%、0.75%、1.00%的壳聚糖),在循环水族箱中饲养草鱼70d。实验结束后测定试验鱼血清中的甲状腺激素(T3和T4)含量和非特异性免疫功能。结果表明,饲料中添加甲状腺激素对草鱼血清中甲状腺激素含量和非特异性免疫功能(头肾和脾脏溶菌酶活性、头肾NO含量)有显著影响(P〈0.05),且两者变化趋势相似。故认为甲状腺激素能够调控草鱼的非特异性免疫功能。壳聚糖对草鱼血清中甲状腺素含量和非特异性免疫功能(吞噬活性、头肾和脾脏溶菌酶活性、头肾NO含量)也有显著影响(P〈0.05),而且在各实验组有较为相似的变化趋势。综合以上结果认为,壳聚糖可以通过内分泌途径甲状腺激素水平的变化实现对草鱼非特异性免疫功能的调控。鉴于0.50%壳聚糖组具有最强的非特异性免疫功能,认为在本实验条件下,壳聚糖在草鱼饲料中的适宜添加量为0.50%。  相似文献   

11.
Anguilla luzonensis and A. huangi were each described in 2009 using eels obtained from northern Luzon Island. We examined the taxonomic status of these two groups of eels using morphological and molecular genetic characters. There were no significant differences in two vertebrae counts between eels of A. luzonensis and A. huangi. Mitochondrial 16S ribosomal RNA and cytochrome b genes sequences were obtained and compared among 28 specimens of A. luzonensis, the holotypes of A. luzonensis and A. huangi, and one specimen of the other 15 anguillid species. The specimens of A. luzonensis exhibited almost identical sequences, including the holotype, with only a few site differences, and the genetic difference between the holotypes of A. luzonensis and A. huangi was within the range of differences of specimens of A. luzonensis. The other anguillid species were genetically very different from A. luzonensis and A. huangi, although A. interioris is a closely related species. It is clear that A. luzonensis and A. huangi are the same species, and according to the principle of priority in zoological nomenclature, A. luzonensis Watanabe, Aoyama, and Tsukamoto, 2009 is the valid species name, and A. huangi Teng, Lin, and Tzeng, 2009 is a junior synonym of A. luzonensis.  相似文献   

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Mosquitofish, Gambusia sp., have been spread throughout the world to biologically control mosquitoes. However, the fish has gained a reputation as an invasive species and has been implicated in displacing native aquatic species. Gambusia affinis are native to the southeastern United States and commonly occur in commercial channel catfish, Ictalurus punctatus, production ponds. We investigated effects of mosquitofish presence on zooplankton populations, water quality, disease occurrence, and fish production in experimental ponds. There were no differences between ponds with or without mosquitofish in numbers of calanoid copepods, cyclopoid copepods, total copepods, Bosmina sp., Ceriodaphnia sp., Moina sp., Daphnia sp., or total cladocerans. There were also no differences in copepod and cladoceran sizes. Copepod nauplii were more numerous during the summer months in ponds with mosquitofish. There were no differences in water quality variables (soluble reactive phosphorus, nitrate, nitrite, ammonia, total nitrogen, total phosphorus, pH) or phytoplankton density between ponds stocked with and without mosquitofish. Catfish production and disease occurrence were also similar between ponds with and without mosquitofish. Although mosquitofish may cause problems when stocked outside their native range, there does not appear to be any adverse effects of mosquitofish presence in catfish production ponds.  相似文献   

15.
Mulloway (Argyrosomus japonicus) occur in estuarine and coastal waters surrounding Australia, Africa, India, Pakistan, China, Korea and Japan, where they are important in fisheries. This study identified that mulloway in south-eastern Australia had similar growth rates, but matured at smaller lengths and younger ages, to those in South Africa and Western Australia. Growth of both sexes was similar to about 5 years, after which females grew faster and attained a greater maximum length than males. Female mulloway matured at 4–5 years of age with a L50 of 68 cm, whereas males matured at 2–3 years of age with a L50 of 51 cm. The commercial fishery in New South Wales was characterised by declining catches and a reduction in the proportion of mulloway of mature lengths in landings. During 2002–2005 commercial landings were dominated (83%) by fish within 15 cm of the current minimum legal total length of 45 cm and aged 2 and 3 years (>80%), even though mulloway can attain lengths of 200 cm and live >30 years. Estimates of the rates of instantaneous total mortality ranged between 0.34 and 0.45, whilst the rate of instantaneous natural mortality (M) was estimated to be approximately 0.12. Yield-per-recruit analyses indicated that mulloway in New South Wales are being growth overfished and substantial increases in yield could be achieved by increasing the length at first harvest. Values of the spawning potential ratio were below 0.2 under a range of mortality estimates, suggesting that mulloway are at risk of recruitment overfishing. These results suggest that the spawning stock of mulloway in south-eastern Australia has been depleted and that remedial management action is required to protect this iconic species.  相似文献   

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Fisheries Science - Fermentation of kamaboko in koji with wheat (KW), rice (KR), brown rice (KB), or soybean (KS) was performed by simulation of tofuyo processing. Moisture and protein content...  相似文献   

18.
The objectives of this study were to evaluate the effects of varying dilutions, pH, temperature, osmolality, and cations on sperm motility parameters in waigieu seaperch, Psammoperca waigiensis. The maximum velocity of average in path (VAP), percentage of motile cells (MOT), and duration of sperm motility (DSM) were observed when semen was diluted in artificial seawater (ASW) at a ratio of 1:100 (144.9 ± 0.6 µm/sec, 95.6 ± 0.4%, and 230.3 ± 2.3 sec, respectively), at 30 C (142.0 ± 0.6 µm/sec, 93.6 ± 0.4%, and 238.3 ± 0.9 sec, respectively), and pH 8 (144.8 ± 0.6 µm/sec, 93.3 ± 0.4%, and 234.0 ± 1.5 sec, respectively). Maximum VAP, MOT, and DSM were obtained in each solution containing 0.6 M NaCl (143.8 ± 1.0 µm/sec, 91.3 ± 2.0%, and 230.6 ± 4.2 sec, respectively), 0.6 M KCl (135.1 ± 3.1 µm/sec, 91.1 ± 3.1%, and 230.3 ± 3.7sec, respectively), 0.2M CaCl2 (105.3 ± 4.7μm/sec, 47.9 ± 2.7%, and 120.7 ± 1.3 sec, respectively), 0.2 M MgCl2 (107.3 ± 3.0 m/s, 42.1 ± 3.3%, and 120.3 ± 4.8 sec, respectively), and osmolality of 400 mOsm/kg (145.1 ± 2.5 µm/sec, 93.0 ± 2.1%, and 346.5 ± 4.4s, respectively). We used these mediums as artificial insemination media for fertilizing matured eggs. The results showed that the fertilization and hatching rates in 0.6 M NaCl (75.3 ± 0.6% and 57.0 ± 2.4%, respectively), ASW (70.8 ± 1.2% and 51.2 ± 1.8%, respectively), or 400 mOsm/kg (72.9 ± 1.8% and 55.3 ± 1.6%, respectively) were higher than that in seawater (63.9 ± 1.2% and 39.2 ± 3.9%, respectively). In conclusion, using 0.6 M NaCl, ASW, or 400 mOsm/kg as an artificial insemination medium is effective for fertilizing of waigieu seaperch.  相似文献   

19.
The effects of exposing the eggs of Pacific threadfin and amberjack eggs (AEs) to different concentrations of hydrogen peroxide for 5 min on hatch rate and survival were assessed in a series of experiments using a petri dish model rearing system. Despite significant inter‐batch variation in hatch rate, it was shown that eggs of both species could be safely exposed to up to 11 340 mg L−1 H2O2 for 5 min. Exposure to 34 230 mg L−1 H2O2 for 5 min was shown to be lethal to AEs at a late stage of development. In two further experiments, it was demonstrated that Pacific threadfin eggs were resistant to all tested concentrations of a range of polyvinylpyrrolidone iodine (PVP‐I) concentrations and contact times (up to 1000 mg L−1 PVP‐I for 10 min). The level of bacteria adhering to the eggs of both species was highly variable. Where eggs were heavily colonized (>104 cfu egg−1), hydrogen peroxide concentrations of at least 11 340 mg L−1, or PVP‐I concentrations higher than 500 mg L−1 for 10 min, were required for effective sterilization. In less colonized batches, rinsing in sterile seawater or exposure to lower (550 mg L−1) concentrations of H2O2 was sufficient to result in high apparent levels of surface sterility (<1 cfu egg−1).  相似文献   

20.
Aquaculture International - The world population continues to increase day by day. Therefore, aquaculture needs to grow in order to response the need for protein foods and to cope with the problem...  相似文献   

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