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1.
硫化物是水产养殖过程中常见的水体污染物之一。为探讨水体硫化物胁迫对虾类血细胞的毒性影响,以不同浓度(0.5 mg/L,2.0 mg/L)的硫化物对凡纳滨对虾(Litopenaeus vannamei)进行胁迫,于胁迫后6、12、24和48 h取血淋巴,应用流式细胞术测定血细胞的总数(THC)、活性氧(ROS)含量、一氧化氮(NO)含量以及凋亡率。结果显示,经0.5 mg/L硫化物胁迫48 h后,对虾THC显著下降至11.78×10~6个/mL,为对照组的72.7%(P0.05),ROS含量为对照组的225.2%(P0.05),血细胞凋亡率显著上升至7.42%(P0.05);经2.0 mg/L硫化物胁迫6 h开始,对虾THC呈现显著的下降(P0.05),血细胞ROS含量和凋亡率显著提高(P0.05);表明硫化物胁迫刺激对虾血细胞产生大量ROS,诱导血细胞凋亡,从而导致THC下降,这一过程可能是硫化物导致虾类细胞免疫下降的重要机制;随着硫化物浓度的升高,其细胞毒性作用明显提高。血细胞NO含量在2.0 mg/L硫化物胁迫12和48 h时显著升高(P0.05),推测NO可能在硫化物胁迫防御调控中起着信号因子的作用。  相似文献   

2.
以新鲜虾血细胞为阴性对照,α-羰基氰化氯苯腙处理的虾血细胞为阳性对照,建立以JC-1为荧光标记的测定虾类血细胞线粒体膜电位的流式细胞术方法.将凡纳滨对虾离体血细胞经不同浓度Cd2+ (10-9~10-3 mol/L)处理6h后,应用该方法测定血细胞线粒体膜电位的变化.结果显示10-4 mol/L和10-3 mol/L Cd2+导致血细胞线粒体膜电位显著下降,表明该浓度的Cd2+会破坏虾血细胞线粒体的功能.本研究结果也显示,应用JC-1标记的流式细胞术测定方法适用于虾类血细胞线粒体膜电位的研究.  相似文献   

3.
为探讨镉(Cd)胁迫下凡纳滨对虾(Litopenaeus vannamei)血细胞的分子响应,以不同浓度的Cd2+(0,0.5 mg/L和5 mg/L)进行胁迫,于不同胁迫时间取血淋巴,测定血细胞中Trx 2、Grx 2、Grx 3和MGST 3的基因表达量变化。结果显示,Trx 2、Grx 3和MGST 3表达量在胁迫的中后期显著上调,Grx 2表达量在5 mg/L Cd2+胁迫6 h开始即有显著的升高,并持续至48 h。结果表明在Cd胁迫作用下,对虾血细胞中上调这四个氧化还原与抗氧化相关基因的表达水平,以进行胁迫防御;Grx 2对Cd胁迫较为敏感,在整个胁迫过程发挥作用,Trx 2、Grx 3和MGST 3主要在胁迫的中后期发挥作用;基因的表达响应具有一定浓度效应。  相似文献   

4.
研究了饲料铜离子含量对亚硝酸盐胁迫下斑节对虾(Penaeus monodon)血细胞参数的影响。向斑节对虾投喂不同铜离子添加量(0,10,25,40,55和110 mg/kg)的饲料8周后,进行24 h 的20 mg/L亚硝酸氮胁迫,取样测定对虾的血细胞总数(THC)、血细胞活性氧(ROS)含量和血细胞凋亡率。结果显示,经亚硝酸盐胁迫后,25-55 mg/kg饲料组对虾能保持较高的THC,以及较低血细胞ROS含量和凋亡率;110 mg/kg饲料组对虾的THC最低,血细胞ROS含量和凋亡率均最高。这些结果表明饲料中添加25-55 mg/kg的铜离子能有效降低亚硝酸盐胁迫对血细胞造成的氧化损害,但过量添加(110 mg/kg)反而对血细胞造成更为严重的损害。  相似文献   

5.
镉对凡纳滨对虾离体血细胞的毒性影响   总被引:1,自引:0,他引:1  
对凡纳滨对虾血细胞进行离体镉离子应激,Cd2+浓度为10-3-10-9M,在应激6h后取样,利用流式细胞术(FCM)检测血细胞的活性和活性氧(ROS)含量。结果显示,当Cd2+浓度为10-5M时,血细胞的ROS含量显著升高;当Cd2+浓度为10-3和10-4M时,血细胞的死亡率和ROS含量均显著上升。这些结果表明Cd2+的细胞毒性作用与其浓度相关,Cd2+浓度达到10-5M时即可诱导血细胞ROS的产生,但当Cd2+浓度达到10-4M及以上时,被诱导产生的ROS对细胞造成氧化伤害从而导致细胞死亡。  相似文献   

6.
潘鲁青  谢鹏  岳峰  孙晓华 《水产学报》2010,34(5):726-732
研究了脂多糖(LPS)、多巴胺(DA)对凡纳滨对虾血细胞吞噬、胞吐及信号通路的影响。结果表明,LPS、DA对凡纳滨对虾血细胞数量、吞噬率和胞吐酚氧化酶活力影响显著(P<0.05),LPS、DA与血细胞孵育30min后,高浓度(1~10mg/L或μmol/L)处理组对虾血细胞数量均随作用浓度增大而明显下降,胞吐酚氧化酶活力则显著升高,LPS处理组对虾血细胞吞噬率显著升高,而DA处理组对虾血细胞吞噬率则表现出明显的下降趋势。同时在LPS作用下,分别加入蛋白激酶C(PKC)、酪氨酸蛋白激酶(TPK)抑制剂chelerythrine、genistein后,凡纳滨对虾血细胞吞噬和胞吐作用均受到明显的抑制作用,抑制剂对吞噬的抑制效果为genistein>chelerythrine,对胞吐的抑制效果则为chelerythrine>genistein;在DA作用下,加入蛋白激酶A(PKA)抑制剂H-89后,血细胞吞噬作用得到增强,但对胞吐作用无明显影响,chelerythrine、genistein均抑制对虾血细胞的胞吐作用,抑制剂的作用效果为chelerythrine>genistein,两种抑制剂对吞噬均无显著影响。  相似文献   

7.
重金属对凡纳滨对虾血细胞吞噬活力的影响   总被引:1,自引:0,他引:1  
为研究不同重金属离子对虾类血细胞的免疫抑制作用,凡纳滨对虾(Ltiopenaeus vannamei)血细胞在离体状态下暴露于重金属离子(Cd^(2+)、Hg2+、Cu2+和Zn2+),浓度分别设置为10-3~10-9 M,在胁迫6h后应用流式细胞术测定血细胞的吞噬活力。结果显示,与对照组相比,浓度为10-3~10-5 M的Cd2+和Hg2+、10-3~10-4 M的Cu2+、10-3 M的Zn2+均显著抑制了对虾血细胞的吞噬活力;当浓度为10-3 M时,Cd2+、Hg2+、Cu2+和Zn2+组的吞噬活力抑制率分别为79.6%,82.6%、68.1%和58.7%。这些结果表明,一定浓度的重金属离子对虾类血细胞吞噬活力具有免疫抑制作用,免疫抑制毒性强度依次为:Hg2+>Cd2+>Cu2+>Zn2+,免疫毒性临界浓度分别为:Cd2+和Hg2+10-5 M、Cu2+10-4 M、Zn2+10-3 M。  相似文献   

8.
氨氮胁迫下白斑综合征病毒对凡纳滨对虾的致病性   总被引:1,自引:0,他引:1  
为了评价养殖水环境中氨氮(NH_4-N)对凡纳滨对虾(Litopenaeus vannamei)的危害性,开展了NH_4-N胁迫对凡纳滨对虾感染白斑综合征病毒(WSSV)后的死亡率、WSSV增殖速率和对虾主要免疫相关酶活性影响的实验。在NH_4-N胁迫质量浓度为15.6 mg·L-1,分别注射2×105和2×106个WSSV粒子,结果显示,NH_4-N胁迫下注射2×105个WSSV粒子的凡纳滨对虾第144小时死亡率达到53.3%,显著高于无胁迫组(40.0%)。对虾鳃组织WSSV荧光定量PCR检测结果显示,NH_4-N胁迫下凡纳滨对虾鳃组织内WSSV的增殖加快。此外,免疫相关酶活性结果显示,NH_4-N浓度突变会促使对虾血清中酚氧化酶(PO)、酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性短暂升高后持续降低。由此可见,NH_4-N胁迫会加快WSSV在患病凡纳滨对虾体内的增殖,导致更高死亡率,这可能是因为胁迫造成了对虾免疫相关酶活性降低和抗病原感染能力下降。  相似文献   

9.
氨氮胁迫下凡纳滨对虾运动行为与能量分配模式变化   总被引:1,自引:1,他引:0  
为研究氨氮胁迫下凡纳滨对虾(Litopenaeus vannamei)行为和能量分配的变化,将体质量为(4.89?0.27)g的凡纳滨对虾幼虾分别置于氨氮质量浓度为0.02 mg/L(对照)、1.00 mg/L、2.50 mg/L、5.00 mg/L,水温(28?0.5)℃的水族箱中养殖2周,监测其行为并对其生长、体组分和能量收支进行测定。结果表明,随氨氮浓度升高,凡纳滨对虾白昼的活动频率、游走距离呈先升后降趋势,而夜间则呈逐渐下降趋势,5.00 mg/L组白昼和夜间活动频率及游走距离均显著低于对照组(P0.05);特定生长率及摄食率均呈逐渐下降趋势,处理组显著低于对照组(P0.05);2.50 mg/L和5.00 mg/L组的凡纳滨对虾脂肪含量和能值均显著低于对照组(P0.05);各处理组摄食能量分配于生长的比例随着氨氮浓度的增大而降低,而消耗于代谢的比例随氨氮浓度增大而升高,与对照组相比,5.00 mg/L组的摄食能用于生长比例下降4.57%,用于代谢消耗的比例上升5.70%。上述结果表明,氨氮胁迫下凡纳滨对虾的夜间活动水平明显下降,摄食量减少,能量利用效率降低,生长速度减慢。  相似文献   

10.
亚硝酸氮和副溶血弧菌对凡纳滨对虾部分免疫指标的影响   总被引:2,自引:0,他引:2  
在养殖水体中含不同浓度亚硝酸氮的情况下,用副溶血弧菌浸泡感染的方式对凡纳滨对虾(Litopenaeus vannamzei)进行毒性实验;并检测了凡纳滨对虾在亚硝酸氮和弧菌胁迫下的部分免疫指标的变化,研究亚硝酸氮和副溶血弧菌对凡纳滨对虾免疫力的影响.结果表明,实验Ⅰ:亚硝酸氮浓度为0.75、1.50、3.00 mg/L组分别胁迫健康凡纳滨对虾,对虾的血细胞数、血清酚氧化酶活力、酸性磷酸酶活力、超氧化物歧化酶活力和过氧化氢酶活力均显著低于对照组(0.005 mg/L)(P<0.05);10 d时,0.005、0.75、1.50、3.00 mg/L组的对虾死亡率分别达到0%、16.7%、20.0%、26.7%.实验Ⅱ:亚硝酸氮胁迫同时,用副溶血弧菌感染对虾,与实验Ⅰ相比,对虾的各项免疫指标受到的影响更大,其各项免疫指标与相应的实验Ⅰ相比,均有显著的差异(P<0.05);第10天时,0.005、0.75、1.50、3.00 mg/L对虾死亡率分别达到33.3%、40.0%、46.7%、50.0%.可见亚硝酸氮浓度越高,弧菌对对虾免疫力的破坏性越大,对虾的死亡率也越高.  相似文献   

11.
Growth, immunological and physiological parameters of white shrimp Litopenaeus vannamei reared at different salinity levels (1, 10, 15, 25 and 35 g/L) at stocking density of 214 shrimp/m3 were examined at 1, 30 and 63 days. Results showed that the total haemocyte count (THC) of shrimp decreased with time at all salinity levels, indicating a potential reduction in the resistance of shrimp against pathogens, since a low value of THC indicates a perturbation of the immune system. Glucose and protein values observed in the haemolymph throughout the study indicate that shrimp adapted well to low salinities (1, 10 and 15 g/L). Although of those shrimp reared at 10 g/L only 83.3% survived, at this salinity, shrimp depicted a higher glucose concentration in haemolymph at the beginning and end of the study.  相似文献   

12.
饥饿胁迫对虾夷扇贝几种免疫因子的影响   总被引:2,自引:0,他引:2  
在(17±1)℃条件下,研究了壳长(8.83±0.52)cm的虾夷扇贝在不同饥饿时间(5、10、15、20、30 d)处理后,血液中血细胞总数、血细胞吞噬水平、酸性磷酸酶、过氧化物歧化酶的变化。结果表明,扇贝的血细胞总数随着饥饿时间的延长逐渐降低,饥饿20 d以上的扇贝血细胞数明显降低,与投喂组差异极显著(P<0.01),扇贝血细胞吞噬率和酸性磷酸酶活性呈现出先小幅上升后下降的趋势,饥饿253、0 d与投喂组差异显著(P<0.05),饥饿253、0 d的扇贝过氧化物歧化酶的活性有所升高,饥饿30d,与投喂组差异显著(P<0.05)。试验表明,饥饿超过20 d,扇贝的免疫系统受到较为严重的破坏。  相似文献   

13.
The capacity of reactive oxygen intermediates production upon haemocyte stimulation is one of the most important immunoparameter utilized to assess the health status in cultivated shrimps. In the present study, we compared oxidative stress potential, by measuring the superoxide anion production in three penaeid shrimps: two wild Atlantic species, the pink shrimp Farfantepenaeus paulensis and the white shrimp Litopenaeus schmitti and one cultivated Pacific species, the white shrimp, Litopenaeus vannamei, through the nitro‐blue‐tetrazolium‐reduction assay. We also proposed an optimized experimental protocol for this assay, that produces rapid and consistent results with low levels of basal superoxide anion (O2?) production by unstimulated haemocytes and high levels of this oxygen radical after cell stimulation. Among the different cell elicitors used (zymosan, laminarin, lipopolysaccharide and phorbol myristate acetate), laminarin (β‐1,3‐glucans – 2 mg mL?1) was the most potent cell activator for the haemocytes of all three penaeids and we recommend this immunostimulant to routinely evaluate shrimp respiratory burst. In general terms, the most elevated levels of O2? production, after cell stimulation with microbial components, were detected in L. schmitti. Interestingly, the stimulation profile of the haemocytes of L. vannamei was more similar to F. paulensis, than to L. schmitti, which is more phylogenetically related.  相似文献   

14.
White spot syndrome virus (WSSV) is a serious pathogen of aquatic crustaceans. Little is known about its transmission in vivo and the immune reaction of its hosts. In this study, the circulating haemocytes of crayfish, Procambarus clarkii, infected by WSSV, and primary haemocyte cultures inoculated with WSSV, were collected and observed by transmission electron microscopy and light microscopy following in situ hybridization. In ultra-thin sections of infected haemocytes, the enveloped virions were seen to be phagocytosed in the cytoplasm and no viral particles were observed in the nuclei. In situ hybridization with WSSV-specific probes also demonstrated that there were no specific positive signals present in the haemocytes. Conversely, strong specific positive signals showed that WSSV replicated in the nuclei of gill cells. As a control, the lymphoid organ of shrimp, Penaeus monodon, infected by WSSV was examined by in situ hybridization which showed that WSSV did not replicate within the tubules of the lymphoid organ. In contrast to previous studies, it is concluded that neither shrimp nor crayfish haemocytes support WSSV replication.  相似文献   

15.
The success of penaeid shrimp aquaculture has been limited by endemic and epidemic infectious diseases around the world and shrimp defense mechanisms are a priority for control, prevention, and diagnosis. The lethal dose (LD50) of Escherichia coli lipopolysaccharides was calculated and pathogen injection and dissolved ammonia concentration on immune response were evaluated in southern white shrimp Litopenaeus schmitti. The lethal dose of lipopolysacharides was 3.78 mg kg− 1 body weight. Injection caused changes in phenoloxidase activity in plasma and nitric oxide and total haemocyte counts within the first 24 h. High concentration of dissolved ammonia caused a decrease in haemocytes by 66% within the first 72 h, when compared to the control. Hemagglutination in plasma was not affected by injection or high concentrations of dissolved ammonia. The results showed that white shrimp recognized the lipopolysacharides and responded to this microbial elicitor, as indicated by a variety of immunological indicators and that increased dissolved ammonia affected the number of circulating haemocytes.  相似文献   

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