The effect of dietary Panax ginseng polysaccharide extract on the immune responses in white shrimp, Litopenaeus vannamei

The immunostimulatory effects of orally administered Panax ginseng root or its polysaccharides (GSP) in white shrimp, Litopenaeus vannamei, were investigated in this study. Shrimp were fed a diet containing 0.4 g kg^?1 GSP over a period of 84 days, during which the activities of total superoxide dismutase (T-SOD), catalase (CAT), glutathione peroxidase (GSH-Px), acid phosphatase (ACP), and alkaline phosphatase (AKP), as well as malondialdehyde (MDA) content, and expressions of cytosolic superoxide dismutase (cyt-SOD), CAT, GSH-Px, and peroxiredoxin (Prx) genes were determined in various tissues of the shrimp. Results showed that the shrimp fed the GSP diet had significantly increased ACP and AKP activities in the gills. The GSP-fed shrimp also displayed significantly increased T-SOD and GSH-Px activities in the gills and hepatopancreas of the shrimp; meanwhile there was enhanced CAT activity in the gills, but decreased MDA content in the gills, hepatopancreas and muscle. The mRNA expressions of cyt-SOD, CAT, GSH-Px and Prx were significantly elevated in the gills and hepatopancreas of the shrimp fed the GSP diet for 84 days, compared with that of the control. Therefore, GSP can be used as an immunostimulant for shrimp through dietary administration to increase immune enzyme activity and modify expression of immune genes in shrimp.     

嗜水气单胞菌感染对背角无齿蚌5种免疫相关酶活力的影响

本研究以背角无齿蚌为材料,利用嗜水气单胞菌为诱导刺激物,对背角无齿蚌进行注射感染,在注射后3、6、12 、24、48h分别取血淋巴、肝胰腺,测定其超氧化物歧化酶（SOD）、酚氧化酶 (PO)、酸性磷酸酶 (ACP)、碱性磷酸酶 (AKP) 及过氧化氢酶 (CAT) 的活力,并研究各项酶活力的变化规律。结果表明：除3h肝胰腺外,实验组的SOD活力均不同程度地高于对照组;血清实验组的PO活力开始显著高于对照组,然后降低,而肝胰腺实验组的PO活力持续高于对照组;另外,与对照组相比,实验组中血淋巴与肝胰腺的ACP、AKP和CAT活力在不同的时间段虽然有所增强,但两者之间无显著差异。因此,认为SOD、PO活性可以作为背角无齿蚌免疫抗病功能指标参数,而ACP、AKP及CAT活性能否作为该参数还有待于进一步研究。     

邻苯二甲酸二丁酯(DBP)胁迫对三疣梭子蟹亚急性毒性

研究不同浓度邻苯二甲酸二丁酯(DBP)对三疣梭子蟹相关酶活的影响及组织损伤。设置了三个处理浓度(0.03 mg/L、0.2 mg/L和0.4 mg/L)分别在第0、5、10、15、20 d进行取样。结果表明,随着DBP浓度的增大,三疣梭子蟹死亡率升高。在0.03 mg/L浓度组胁迫下,血清超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和碱性磷酸酶(AKP)活性呈现升高-下降交替波动,过氧化物酶(POD)活性在第10 d低于对照组(P<0.05),其余时间与对照组持平,酸性磷酸酶(ACP)活性在第5 d较对照组显著升高(P<0.05),此后一直保持对照组水平;在0.2 mg/L浓度组胁迫下,血清SOD活性呈现先升高后下降的趋势,POD活性变化不明显,CAT活性先下降后升高,此后恢复到对照组水平,ACP活性在第15 d出现最高值,其余时间与对照组持平,AKP活性呈现先升高后恢复的波动趋势;0.4 mg/L浓度组SOD、CAT和AKP活性呈现先升高后下降的趋势,POD活性和ACP活性在第20 d较对照组低(P<0.05),其余时间与对照组持平。组织切片观察可见,DBP胁迫损伤了三疣梭子蟹肝胰腺和鳃组织,DBP浓度越大,胁迫时间越长,损伤越明显。     

高盐胁迫对凡纳滨对虾消化及免疫相关酶活力的影响

为探讨高盐对凡纳滨对虾(Litopenaeus vannamei)消化及免疫相关酶活力的影响,实验设置了30、40、50、60共4个盐度梯度。对虾体长(7.84±0.68)cm,养殖密度333尾/m~3,每个梯度设3个平行,实验周期30d。取血淋巴、肌肉、肝胰腺等组织,检测其超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、碱性磷酸酶(AKP)和酸性磷酸酶(ACP)及蛋白酶、脂肪酶、淀粉酶活力。结果表明,盐度显著影响凡纳滨对虾肝胰脏中胃蛋白酶、脂肪酶、淀粉酶的活力(P0.05);随着盐度增加,消化相关酶活力均不断下降,处理间差异显著(P0.05);盐度对凡纳滨对虾不同组织的免疫指标产生影响,表现为随着盐度升高,血淋巴中,AKP活力逐渐升高,ACP、CAT和SOD活力均表现为先升高后降低;肌肉中,AKP、ACP和SOD活力呈现先升高后降低的变化趋势;肝胰脏中,AKP活力呈现先降低后升高再降低的变化趋势,ACP活力高盐处理间差异不显著(P0.05),CAT活力先降低后升高,SOD活力盐度40后逐渐降低。实验结果初步说明,高盐显著影响凡纳滨对虾的消化及免疫相关酶活力,且盐度对不同组织中免疫酶活力影响存在一定的组织特异性,50以上的高盐胁迫对对虾消化和免疫相关酶活力的影响尤为显著。     

Effects of dietary live yeast Hanseniaspora opuntiae C21 on the immune and disease resistance against Vibrio splendidus infection in juvenile sea cucumber Apostichopus japonicus

A feeding experiment was conducted to determine effects of Hanseniaspora opuntiae C21 on immune response and disease resistance against Vibrio splendidus infection in juvenile sea cucumbers Apostichopus japonicus. Sea cucumbers were fed with either diets containing C21 at 10⁴, 10⁵ and 10⁶ CFU g^?1 feed or a control diet for 30–50 days, respectively. After feeding for 30 days and 45 days, five sea cucumbers from each tank were sampled for immunological analyses. Results indicated that C21 significantly improved the phagocytic activity in coelomocytes of sea cucumbers (P < 0.05). Moreover, C21 administration significantly enhanced lysozyme (LSZ), phenoloxidase activity (PO), total nitric oxide synthase (T-NOS), superoxide dismutase (SOD), alkaline phosphatase (AKP) and acid phosphatase (ACP) activities in coelomic fluid, and LSZ, T-NOS, AKP and ACP activities in coelomocytes lysate supernatant (CLS) of sea cucumbers (P < 0.05). After feeding for 45 days, 10 sea cucumbers from each dose group were challenged with V. splendidus NB13. Cumulative incidence and mortality of sea cucumbers fed with C21 were found to be lower than those of control group. After feeding for 50 days, sea cucumbers in 10⁴ CFU g^?1 C21 treatment and control tanks were subjected to acute salinity changes (from 30 to 20) for 24 h in the laboratory, and the immunological parameters were measured to evaluate the immune capacities of the A. japonicus. Phagocytic, LAZ and T-NOS activities of C21-treated group were higher than those of control group, indicating that salinity stress tolerance of sea cucumber was enhanced by C21. The present results showed that a diet supplemented with C21 could stimulate the immune system of juvenile A. japonicus thus enhancing their resistance against V. splendidus.     

The immune responses in Chinese mitten crab Eriocheir sinensis challenged with double-stranded RNA

Double-stranded RNA (dsRNA) is a virus-associated molecular pattern which induces antiviral innate immune responses and RNA interference (RNAi) in mammals. In invertebrates, RNAi phenomenon has been widely studied, but dsRNA-induced innate immune response is seldom reported. In the present study, two different dsRNAs specific for green fluorescent protein (GFP) and the putative D1 protein of photosystem II (NoPSD) from Nannochloropsis oculata, were employed to challenge Chinese mitten crab Eriocheir sinensis. The temporal changes of phenoloxidase (PO), acid phosphatase (ACP), superoxide dismutase (SOD) and malondialdehyde (MDA) content, as well as the mRNA expression of some immune-related genes were examined in order to estimate the effect of dsRNAs on the innate immunity of E. sinensis. The activities of PO, ACP and SOD significantly increased after dsRNA treatment, whereas malondialdehyde (MDA) content did not change significantly. Among the examined genes, only the mRNA expression of EsALF, an antibacterial peptide in E. sinensis, was significantly up-regulated (about 5 fold, P < 0.05) at 12 h after dsRNA treatment, while no significant expression changes were observed among the other immune genes. The increase of PO, ACP and SOD activities, and mRNA expression level of EsALF after dsRNA stimulation indicate that phenoloxidase, hydrolytic enzyme, antioxidation and EsALF were involved in dsRNA-induced innate immunity, suggesting that broad-spectrum immune responses could be induced by dsRNA in E. sinensis.     

黑胸散白蚁工蚁与工蚁型补充生殖蚁体内抗氧化酶和解毒酶的活性及基因表达变化

【目的】社会性白蚁的工蚁可以转化成补充生殖蚁参与生殖,提高自身适合度。本研究旨在探究工蚁转变成补充生殖蚁后,与氧化应激抗衰老相关的抗氧化酶和解毒酶活性的变化,为揭示生殖蚁的繁殖和抗衰老机制提供参考。【方法】利用生化方法分别测定黑胸散白蚁Reticulitermes chinensis工蚁、工蚁型补充生殖蚁和原始生殖蚁体内的2种抗氧化酶过氧化氢酶(catalase, CAT)和超氧化物歧化酶(superoxide dismutase, SOD)与4种解毒酶酸性磷酸酶(acid phosphatase, ACP)、碱性磷酸酶(alkaline phosphatase, AKP)、羧酸酯酶(carboxylesterase, CarE)和细胞色素P450(cytochrome P450, CYP450)的酶活性;同时利用qRT-PCR检测这些酶对应的基因RsCAT, RsSOD, RsACP, RsCYP450和RsCarE的表达量。【结果】与雌性工蚁相比,黑胸散白蚁工蚁型补充生殖蚁体内的CAT, SOD, ACP, AKP和CarE活性显著上升,分别达到雌性工蚁的5.82, 1.41, 1.39, 2.27和2.70倍,CYP450活性在两品级间没有显著差异;RsCAT, RsSOD, RsACP, RsCarE和RsCYP450的表达量也显著增加。补充生殖蚁体内CAT和ACP活性显著高于原始生殖蚁的,而SOD和AKP活性在两品级间没有显著差异;雌雄补充生殖蚁RsCAT的相对表达量分别是原始生殖蚁的5.68和3.60倍,RsACP的相对表达量分别是原始生殖蚁的81.12和46.72倍。【结论】黑胸散白蚁工蚁由非生殖品级转化为生殖品级以后,体内抗氧化酶和解毒酶的酶活力和基因表达水平显著升高,从一定程度上揭示了生殖品级的抗衰老机制。     

盐度渐变与骤变对脊尾白虾渗透、代谢及免疫相关酶活力的影响

为探讨盐度变化对脊尾白虾(Exopalaemon carinicauda)渗透、代谢及免疫相关酶活力的影响,实验设置了盐度渐变和骤变两个实验。渐变实验,设置5、10、15、20、25、30、33(CK)、40和45共9个盐度梯度;骤变实验,盐度从33突变至0、5、15、25和45,检测血清ATP酶(包括Na+/K+-ATP酶和总ATP酶)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)及超氧化物歧化酶(SOD)活力。结果表明,渐变情况下,盐度为5时,ATP酶活力出现最高值,然后随着盐度的升高表现出先降低后升高的趋势。总ATP酶活力在盐度为15—30之间较稳定,并在此范围内达到最低值。AKP和ACP活力几乎不受盐度渐变的影响。SOD活力随盐度的升高,先上升后下降,并在盐度为33时达到最大值。骤变情况下,ATP酶活力随时间波动较大,AKP和SOD随时间波动较小,而ACP几乎不受影响。结果说明,盐度骤变对脊尾白虾酶活力的影响较盐度渐变明显,ATPase和SOD活力更易随盐度而变化,代谢酶(AKP、ACP)受盐度变化的影响较小,说明渗透调节和免疫相关酶活力对盐度变化反应敏感,养殖过程中应尽量保持盐度稳定。     

镉对长江华溪蟹免疫相关酶活性的影响

采用浸浴法研究了镉(Cd2+)对长江华溪蟹Sinopotamon yangtsekiense免疫系统相关酶的影响.Cd2+处理组浓度分别为7.25 mg/L、14.5 mg/L、29mg/L、58 mg/L和116 mg/L,同时设对照组.分别在Cd2+处理24 h、48h、72 h和96 h取血清进行酚氧化酶(PO)、超氧化物歧化酶(SOD)、酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活力测定.结果显示,在相同处理时间,随着cd2+浓度增加,PO和SOD活力均呈现先升后降的趋势,且分别于24 h、7.25 mg/L和96 h、14.5 mg/L时达到最大值.当处理浓度相同、作用时间延长时,ACP活力先升后降,且在48 h、58 mg/L时升至最高,AKP活力则持续上升.表明Cd2+对长江华溪蟹免疫相关酶活力具有不同程度的影响,且其作用和效果具有浓度及时间效应.PO和SOD活力变化能够灵敏反映Cd2+对水生动物的胁迫程度,可作为指示镉污染的生物学指标.     

Immunomodulatory effect of Withania somnifera supplementation diet in the giant freshwater prawn Macrobrachium rosenbergii (de Man) against Aeromonas hydrophila

The effect of Withania somnifera extract supplementation diets on innate immune response in giant freshwater prawn Macrobrachium rosenbergii (de Man) against Aeromonas hydrophila was investigated. The bacterial clearance efficiency significantly increased in prawn fed with 0.1% and 1.0% doses of W. somnifera supplementation diet against pathogen from weeks 1-4 as compared to the control. The innate immune parameters such as, phenoloxidase activity, superoxide anion level, superoxide dismutase activity, nitrate, and nitrite concentrations were significantly enhanced in prawn fed with 0.1% and 1.0% doses of W. somnifera supplementation diet from weeks 1-4 against pathogen. The total hemocyte counts (THC) significantly increased in prawn fed with 0.1% and 1.0% doses diet from weeks 1-4 against pathogen as compared to the control. These results strongly suggested that administration of W. somnifera through supplementation diet positively enhances the innate immune system and enhanced survival rate in M. rosenbergii against A. hydrophila infection.     

氨氮对中华小长臂虾的急性毒性及非特异性免疫指标的影响

采用生物毒性实验方法研究了氨氮对中华小长臂虾的急性毒性作用, 结果表明: 在温度为(18±1)℃, pH为7.3±0.1条件下, 氨氮对中华小长臂虾24h、48h、72h、96h 的半致死浓度(LC₅₀)分别为565.47、371.16、291.16和272.50 mg/L, 安全浓度为 27.25 mg/L。转化为非离子氨的LC₅₀分别为3.74、2.45、1.93 和1.80 mg/L, 安全浓度为0.18 mg/L。根据96h 的LC₅₀和安全浓度按照等差数列设置5个氨氮浓度梯度, 分别60、100、140、180和220 mg/L, 研究氨氮胁迫对中华小长臂虾非特异性免疫指标的影响。结果显示: 在24h时, 除了220 mg/L的肌肉组织, 中华小长臂虾肝胰腺和肌肉中的超氧化物歧化酶(SOD)活性显著性高于对照组, 并具有明显的剂量效应, 在48—96h均回落到正常水平; 在24h时, 中华小长臂虾氨氮处理组中肝胰腺的酸性磷酸酶(ACP)与对照组未发生显著变化, 而碱性磷酸酶(AKP)则显著高于对照组, 在48—96h两者的140、180和220 mg/L处理组均显著低于对照组; 除了140 mg/L 处理组的ACP活性外, 肌肉中的ACP和AKP活性从24h开始就出现了显著性下降, 始终低于对照组。研究获得了氨氮对中华小长臂虾的急性毒性结果和在高氨氮胁迫下非特异性免疫指标的变化规律, 发现中华小长臂虾对氨氮具有较强的耐受性, 但高浓度的氨氮会对中华小长臂虾的免疫酶活性产生抑制作用, 研究结果可为中华小长臂虾健康养殖发展提供科学依据。     

取食感染大麦黄矮病毒的小麦后介体麦长管蚜和非介体禾谷缢管蚜体内抗氧化酶和解毒酶活性的变化

【目的】小麦黄矮病［病原为大麦黄矮病毒(barley yellow dwarf virus, BYDV)］是危害小麦生产的主要病害之一,GAV是BYDV在我国的主流株系,且其在田间与介体麦长管蚜 Sitobion avenae 和非介体禾谷缢管蚜Rhopalosiphum padi同时发生,对小麦产量造成严重影响。本研究旨在探究大麦黄矮病毒胁迫下介体和非介体蚜虫体内重要保护酶和解毒酶活性变化规律,为揭示病毒 蚜虫互作生理生化机理提供参考。【方法】利用生化方法测定取食健康小麦(空白对照组),取食前期经无毒麦二叉蚜Schizaphis graminum 3龄若蚜为害72 h的小麦(条件对照组)及取食前期经携带BYDV-GAV麦二叉蚜3龄若蚜为害72 h的小麦(处理组)7 d后,介体麦长管蚜与非介体禾谷缢管蚜成蚜体内过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)等重要保护酶及乙酰胆碱酯酶(AchE)、酸性磷酸酶(ACP)和碱性磷酸酶(AKP)等重要解毒酶活性变化。【结果】取食前期经BYDV-GAV感染的麦二叉蚜3龄若蚜为害的小麦7 d后,介体麦长管蚜成蚜体内POD, SOD和AKP活性比空白对照组显著上升,但与条件对照组相比无显著差异;且条件对照组比空白对照组显著上升。取食经BYDV-GAV感染的麦二叉蚜3龄若蚜为害的小麦7 d后,非介体禾谷缢管蚜成蚜体内SOD,AKP和AchE活性比空白对照组显著下降,ACP活性显著上升,但与条件对照组相比仅ACP活性显著上升;且条件对照组禾谷缢管蚜体内AKP和AchE比空白对照组显著下降,ACP活性显著上升。【结论】取食感染BYDV-GAV小麦后介体麦长管蚜成蚜体内POD, SOD和AKP酶活性升高是前期蚜虫的为害造成,与BYDV-GAV无关;非介体禾谷缢管蚜成蚜体内AKP和AchE酶活性下降是前期蚜虫为害引起,SOD酶活性下降是前期蚜虫为害和BYDV-GAV的综合作用,而BYDV-GAV和前期蚜虫为害均能使禾谷缢管蚜体内ACP酶活性上升。     

基于原核表达的副溶血性弧菌类毒素疫苗对海水鱼类免疫保护作用

目的:实现副溶血性弧菌溶血毒素基因tdh克隆与表达,并以表达产物TDH作免疫原,研究其对海水鱼免疫活性的影响.方法:PCR扩增tdh;构建重组质粒(pET-28-TDH);IPTG诱导表达;SDS-PAGE检测:以纯化的TDH融合蛋白脱毒为类毒素免疫原免疫健康真鲷,测定其血清中过氧化氢酶(CAT)、酸性磷酸酶(ACP)、超氧化物歧化酶(SOD)和碱性磷酸酶(AKP)活性变化;用副溶血性弧菌病原攻毒,检测免疫保护作用.结果:tdh基因成功克隆并表达;表达蛋白相对分子量为21 kDa,证实为TDH蛋白;类毒素免疫真鲷后,其血清中总超氧化物歧化酶(T- SOD)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)和过氧化氢酶(CAT)活力在注射后48h、24h、24h、2Ah达到最高,最高点分别高于对照组77％、328％、75％、381％;72 h后均恢复至对照组水平.结论:类毒素对真鲷免疫系统具有明显的刺激作用;类毒素免疫后对攻毒的真鲷保护率达50％.     

急性低氧/复氧胁迫对克氏原螯虾抗氧化-能量代谢的影响

为研究低氧/复氧胁迫对克氏原螯虾(Procambarus clarkii)抗氧化及能量代谢的影响,将克氏原螯虾暴露于(1.0±0.1) mg/L急性低氧胁迫和后续(6.8±0.2) mg/L复氧环境中,于低氧胁迫1h、6h及复氧1h、12h分别采集肝胰腺、鳃和血淋巴,研究低氧/复氧胁迫下克氏原螯虾抗氧化-能量代谢酶的活力变化,分析鳃和肝胰腺组织的超微结构改变。在低氧胁迫下,肝胰腺和血淋巴中SOD酶活力显著下降(P<0.05);复氧以后,肝胰腺、血淋巴及鳃组织中SOD酶活力均出现了显著上升(P<0.05)。SOD酶活力变化可能与复氧过程中超氧阴离子自由基的过量产生有关。在复氧12h后,血淋巴和鳃组织中MDA含量均出现了显著性增加(P<0.01),提示机体细胞在复氧胁迫下产生了脂质过氧化。在低氧胁迫下,肝胰腺、鳃和血淋巴中ACP、AKP酶活力显著上升(P<0.05);在复氧12h后,肝胰腺和鳃组织中ACP酶活力显著降低(P<0.01)。显示低氧/复氧胁迫影响了机体的非特异性免疫应答。在急性低氧胁迫下,肝胰腺、血淋巴与鳃组织中的LDH含量和总ATPase活力均显...     

Antioxidant response and oxidative stress levels in Macrobrachium borellii (Crustacea: Palaemonidae) exposed to the water-soluble fraction of petroleum

The aim of the present work was to evaluate the effect of the water soluble fraction of hydrocarbons (WSF) on the antioxidant status of the freshwater prawn Macrobrachium borellii. First, seasonal variations were studied in a non-polluted area. Hepatopancreas and gills showed season-related fluctuations in catalase (CAT), glutathione-S-transferase (GST) activities and in lipid peroxidation levels (LPO), but not in superoxide dismutase (SOD). Then, adults were exposed semi-statically to sublethal doses for 7days. CAT, SOD, GST, and glutathione peroxidase (GPx) activities and LPO, reduced glutathione (GSH) and protein oxidation (PO) levels were determined. Exposed individuals showed significant increases in CAT, SOD, and GST activities in hepatopancreas and CAT activity in gills. GPx activity did not vary in either tissues. While LPO levels increased, GSH levels decreased significantly in hepatopancreas of exposed animals, but PO levels showed no variation. Induction of SOD was also assessed by Real-time PCR mRNA expression in hepatopancreas. The non-enzymatic antioxidant activity was also tested; ABTS 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) was higher in hemolymph of treated-prawns compared to controls, but ferric reducing activity of plasma assay (FRAP) values did not change. Taken together, the present results indicated that the antioxidant defenses of M. borellii, mainly in hepatopancreas, were significantly affected by aquatic hydrocarbon contamination, regardless of the season.     

长江口中华绒螯蟹放流亲蟹对环境的生理适应

按回捕日期取样测定长江口中华绒螯蟹亲蟹标志放流前后血清与肝胰腺的生理指标, 初步研究了放流亲蟹对长江口环境的生理适应过程。结果表明, 在放流后6d, 亲蟹肝胰腺超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性以及血清甘油三酯(TG)和血蓝蛋白含量均降低, SOD、CAT和TG在放流后9d达到较低水平, 其中SOD活性和TG含量较放流前显著降低(P<0.05); 肝胰腺酸性磷酸酶(ACP)以及血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)、总蛋白质(TP)、白蛋白(ALB)、总胆固醇(TC)和肌酐(CREA)水平在放流后总体呈现先升高后降低的趋势; 在放流后79d, 肝胰腺SOD、CAT、ACP与血清ALP、ALT、AST活性均恢复至放流前水平(P>0.05), 而血清TP、血蓝蛋白、ALB、TC、TG、CREA含量较放流前均显著降低(P<0.05)。综合各项指标表明, 中华绒螯蟹亲蟹在放流后6d内出现免疫力下降、代谢增强等反应, 放流22d后亲蟹各项机能逐步恢复, 并在70d后接近或达到放流前水平。建议今后放流前对亲蟹进行环境适应性驯化, 以便提高亲蟹增殖放流效果。     

两种鼠蚤在新羽化和吸血后不同时间三种酶的组织化学研究

采用组织化学技术,研究了不等单蚤Monopsyllus anisus (Rothschild)和缓慢细蚤Leptopsylla segnis (Schonherr)新羽化和吸血后24 h、48 h和72 h碱性磷酸酶、酸性磷酸酶和三磷酸腺苷酶的分布和活性。显微摄影及定量图像分析结果显示：新羽化蚤碱性和酸性磷酸酶主要存在于中肠、神经细胞核、精子束头部、精巢附腺、射精管、输卵管和受精囊附腺中;三磷酸腺苷酶各组织中均有分布。吸血消化后,两种蚤中肠3种酶活性均有增强;除碱性磷酸酶在消化72 h 后酶活性有所下降外,其余不同消化时间酶活性增强程度上不存在显著差异。两种蚤卵母细胞发育成熟过程中3种酶活性亦见增强。     

重金属镉对鲫鱼碱性磷酸酶和酸性磷酸酶活性的影响

研究了重金属镉对鲫鱼肠、肝胰脏、鳃组织碱性磷酸酶和酸性磷酸酶活性的影响。结果表明,在0.2、0.4、0.8mg/L镉浓度条件下静态染毒12h、24h、48h、96h后,鲫鱼肠、鳃组织中碱性磷酸酶(AKP)和酸性磷酸酶(ACP)的活性降低,肝和胰脏的碱性磷酸酶活性没有明显变化,其酸性磷酸酶活性则升高。     

Study on immune regulation in Hyriopsis cumingii Lea: Effect of pearl-nucleus insertion in the visceral mass on immune factors present in the hemolymph

This study was aimed at elucidating the mechanism of immune responses in fresh water mussels during pearl culture. Alpha-2 macroglobulin gene (α₂M) expression, acid phosphatase (ACP) and superoxide dismutase (SOD) activities, and hemocyte counts were evaluated after inserting a pearl nucleus into the visceral mass of Hyriopsis cumingii Lea (H. Lea). We selected 60 H. Leas and randomly assigned them to 2 groups (each group contained 3 replicates of 10 individuals), and individuals among one group were treated by inserting pearl nucleus into the visceral mass. Real-time quantitative polymerase chain reaction (Q-PCR) was used to evaluate α₂M gene expression, and the activities of ACP and SOD in hemocytes and serum were determined after 1, 2, 3, 5, and 10 days after nucleus insertion. Hemocyte morphologies and numbers on the 5th day after insertion were studied using phase-contrast microscope (PCM), optical microscope and flow cytometry (FCM). All observations suggested that the insertion of the pearl nucleus in the visceral mass had a significant effect on α₂M gene expression, ACP and SOD activities, and hemocyte characteristics. The α₂M gene expression was sharply up-regulated on the 3rd day after nucleus insertion, and it was significantly higher in the test groups on the 3rd, 5th, and 10th days than those in the control groups (P < 0.05). On the 1st to 3rd after treatment, ACP activity in the test group was higher than that in the control group (P < 0.05). SOD activity in the serum was remarkably higher in the test groups than in the control group, and exhibited significant differences on the 3rd, 5th, and 10th days (P < 0.05). However, the SOD activity in hemocytes was lower in the test group than in the control group, and it exhibited significant differences on all days, except on the 3rd day (P < 0.05). The hemocytes were divisible into 2 types: granulocytes (GR) and hyalinocytes (HY). The hemocyte morphology, protuberances, vesicle-like bodies, and density increased significantly (P < 0.05), and the number of GR increased, while that of HY decreased after nucleus insertion. These results indicated that the insertion of pearl nucleus enhanced the immune response in H. Lea.