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。     

投喂频次对凡纳滨对虾行为及免疫功能的影响

对凡纳滨对虾在每日不同投喂频次(1次/d、2次/d、6次/d),每次过量投饵条件下的行为、生长和免疫指标进行了研究.结果显示:随着投喂频次增加,凡纳滨对虾的活动水平和昼夜游走距离均呈逐渐升高的趋势,特定生长率显著升高.在白昼,6次·d^-1组对虾的活动频率显著高于其它组(p<0.05);在夜间,2次·d^-1组的活动频率显著高于其它组(p<0.05).1次·d^-1组对虾的酚氧化酶PO、过氧化氢酶CAT和超氧化物歧化酶SOD活力下降,丙二醛MDA含量升高;2次·d^-1组凡纳滨对虾的CAT活力、SOD活力和MDA含量在实验期间趋于稳定;6次·d^-1组免疫酶活力呈缓慢升高的态势.     

重金属离子对凡纳滨对虾肝胰脏、鳃丝和血液SOD活力的影响

研究了3种重金属离子(Cu²⁺、Zn²⁺、Cd²⁺)在96 h内对凡纳滨对虾(Litopenaeus vannamei)对肝胰脏、鳃丝和血液超氧化物歧化酶(SOD)活力的影响.结果表明,凡纳滨对虾SOD活力在3种重金属离子作用下随取样时间变化显著(P＜0.0),Cu²⁺在实验浓度范围内(0.1～1 mg·L^-1),肝胰脏、鳃丝和血液的SOD活力随时间延长呈一峰值变化,Zn²⁺在10 mg·L^-1时对肝胰脏表现为显著抑制作用,Cd²⁺在0. mg·L^-1时对肝胰脏和鳃丝起显著抑制作用,0.2 mg·L^-1对鳃丝SOD活力无显著变化(P＞0.0),其他浓度Zn²⁺(＜10 mg·L^-1)、Cd²⁺(＜0.2 mg·L^-1)对各组织器官SOD活力的影响随时间延长均呈现先升高后下降的趋势.3种重金属离子对凡纳滨对虾肝胰脏、鳃丝、血液SOD活力的影响呈现明显的剂量-时间效应关系.其SOD活力大小顺序为肝胰脏＞鳃丝＞血液,3种重金属离子对凡纳滨对虾伤害大小顺序为Cd²⁺＞Cu²⁺＞Zn²⁺.     

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。     

绿原酸对凡纳滨对虾抗氧化系统及抗低盐度胁迫的影响

对绿原酸调节凡纳滨对虾(Litopenaeus vannamei)血淋巴抗氧化系统功能及抗低盐度胁迫的效果进行了评价。360尾凡纳滨对虾随机分为4组,分别投喂含有0、100、200和400 mg/kg绿原酸的饲料28 d,随后将对虾从盐度为32的天然海水直接转入至盐度为10的水中胁迫72 h。结果表明,在正常养殖条件下,绿原酸对凡纳滨对虾的成活率、血淋巴总抗氧化能力(Total antioxidative capacity, T-AOC)、超氧化物歧化酶(Superoxide dismutase, SOD)及过氧化氢酶(Catalase, CAT)活力均无明显影响,然而投喂含有绿原酸的饲料14 d,对虾血淋巴谷胱甘肽过氧化物酶(Glutathione peroxidase, GPx)活性和血淋巴细胞GPx和CAT基因表达水平均显著高于对照组(P<0.05);低盐度胁迫24 h,绿原酸组凡纳滨对虾的存活率较对照组提高10%,但各组之间无显著性差异(P>0.05);低盐度胁迫24 h,各组凡纳滨对虾血淋巴T-AOC、SOD和GPx活性与胁迫前相比均显著增加,说明低盐度胁迫条件下机体产生了抗氧化胁迫反应,同时绿原酸组对虾血淋巴GPx、CAT活性均显著高于对照组(P<0.05);低盐度胁迫72 h,绿原酸组对虾血淋巴T-AOC、GPx和CAT活性和血淋巴细胞GPx和CAT基因表达水平均明显高于对照组。上述结果表明绿原酸可有效调节凡纳滨对虾的抗氧化系统功能,增强对虾对于低盐度胁迫下的适应能力。     

虾头提取物对凡纳滨对虾生长和免疫因子的影响

以添加不同量虾头提取物(0、1.0%、1.5%、2.0%、2.5%、3.0%)的饲料投喂初体质量为6.35±0.38 g的凡纳滨对虾49 d,研究其对对虾的生长及免疫因子的影响。各组凡纳滨对虾的生长及体成分组成没有显著差异。与对照组相比,1.5%~3.0%添加量组对虾血清中的ACP活力明显提高;2.0%~3.0%添加量组对虾血清中的AKP活力明显提高;各添加量组对虾血清中的PO活力都有明显提高。实验结果表明,添加适当剂量的虾头提取物可以增强凡纳滨对虾的免疫能力。     

饲料中铜水平对凡纳滨对虾免疫相关基因表达和抗菌能力的影响

在饲料中添加0、30和50 mg Cu/kg饲料的蛋氨酸铜,投喂凡纳滨对虾(Litopenaeus vannamei)7、14和21d,检测对虾体组织铜蓄积、免疫相关基因(Toll受体mRNA和溶菌酶mRNA)表达水平和免疫抗菌能力的变化。结果表明:凡纳滨对虾肝胰腺铜含量随着饲料蛋氨酸铜添加量增加及投喂时间延长而显著增加(P0.05);对虾肌肉的铜含量显著低于肝胰腺的铜含量。饲料中铜水平对凡纳滨对虾肌肉、血淋巴及肝胰腺中溶菌酶活性无显著影响(P0.05)。对虾组织SOD活性因饲料中铜水平和投喂时间变化显著,添加30 mgCu/kg组对虾肌肉、血淋巴和肝胰腺中SOD活性在第21天时显著高于其他两组(P0.05)。饲料中铜水平对凡纳滨对虾鳃组织中溶菌酶mRNA表达水平无显著影响,但显著影响鳃组织Toll受体mRNA表达水平(P0.05)。第7天时凡纳滨对虾Toll受体mRNA表达水平随着饲料铜水平升高而显著升高(P0.05);第14和第21天时,Toll受体mRNA表达水平在摄食添加30 mg Cu/kg组最高。人工急性感染溶藻弧菌(Vibrioalginolyticus)实验表明,第7天时,摄食添加50 mg Cu/kg组凡纳滨对虾全致死时间和半致死时间长于未添加铜组和添加30 mgCu/kg组,但在第14天,摄食添加30 mg Cu/kg组的全致死时间和半致死时间最长。研究表明饲料铜添加水平不但影响组织中铜的蓄积,还影响凡纳滨对虾SOD活性和Toll受体mRNA表达水平,从而影响机体的抗弧菌能力。     

亚硝酸盐氮对凡纳滨对虾毒性和抗病相关因子影响

用常规生物毒性实验方法,在不同盐度下进行亚硝酸盐氮对凡纳滨对虾的急性毒性实验;并加亚硝酸盐氮于凡纳滨对虾的养殖环境中,检测与抗病力相关因子的变化。研究亚硝酸盐氮对凡纳滨对虾的毒性和抗病力相关因子的影响。结果表明:盐度对亚硝酸盐氮的毒性有较大影响。盐度为31时,24hLC50、48hLC50、72hLC50和96hLC50分别为314.9mg/L1、75.3mg/L1、00.2mg/L和89.0mg/L;盐度为17时,分别为132.3mg/L、65.6mg/L、51.3mg/L和39.5mg/L。盐度31实验组的亚硝酸盐氮半致死浓度均显著(P<0.05)高于盐度17实验组。在低盐度条件下亚硝酸盐氮的毒性较强。亚硝酸盐氮对凡纳滨对虾抗病力相关因子有显著的影响。亚硝酸盐氮浓度为4.0mg/L和8.0mg/L时,其血细胞数、超氧化物歧化酶(SOD)活力、酚氧化酶(PO)活力、抗菌活力(Ua)、溶菌活力(UL)和血清蛋白含量均显著(P<0.05)低于对照组;不吸污组抗病力相关因子活性均显著(P<0.05)低于吸污组。低浓度的亚硝酸盐氮可降低凡纳滨对虾抗病能力,亚硝酸盐氮浓度越高,其抗病能力越弱。     

盐度对凡纳滨对虾体组织蛋白质积累、氨基酸组成和转氨酶活性的影响

本文研究了低、中和高三个盐度水平(分别为3‰、17‰和32‰)对凡纳滨对虾(Litopenaeus vannamei)各组织蛋白质的积累、肌肉谷草转氨酶和谷丙转氨酶活力、肌肉总氨基酸和游离氨基酸组成和含量的影响。结果显示,经过50d不同盐度水平的试验,低盐度组对虾的肝胰腺和血淋巴中可溶性蛋白质含量显著高于中、高盐度组(p<0.05),而肌肉中可溶性蛋白质含量在各处理组间无显著性差异;低、高盐度均导致肌肉中谷丙转氨酶和谷草转氨酶活力升高,但是各处理间的差异不显著;低、高盐度组凡纳滨对虾肌肉总氨基酸和总必需氨基酸含量均显著高于中盐度组(p<0.05),中、低盐度处理组非必需氨基酸含量差异不显著,而低盐度组对虾肌肉中蛋氨酸、丝氨酸、半胱氨酸和脯氨酸含量均显著低于中盐度组(p<0.05),其中脯氨酸为常见的5种主要渗透调节氨基酸之一;低、高盐度组对虾肌肉总游离氨基酸含量显著高于中盐度组(p<0.05),而盐度对机体绝大部分肌肉游离氨基酸含量的影响不显著(p>0.05)。结果显示,当环境盐度偏离凡纳滨对虾最适生长盐度时,其可通过在肝胰腺和血淋巴蛋白质积累及提高自身转氨酶活力,来获得机体在渗透调节供能时所需的氨基酸,而这些氨基酸以脯氨酸为主。     

斑节对虾血淋巴免疫指标变化与其养殖池塘理化因子的关系

2009 年8 月至9 月期间, 在广东粤西选择2 口斑节对虾养殖土池(303、501 号池), 每14 天一次定期采集养殖对虾和水样, 检测了对虾血清的PO、SOD、Ua、AKP、ACP 等免疫酶活力和水温、DO、氨氮、亚硝氮等环境因子, 探讨养殖对虾免疫指标与环境因子的相关关系。结果发现, 较高的COD、BOD、pH 与较低盐度的303 号池对虾血清SOD、Ua、AKP、ACP活力大体上低于501 号池, PO 活力高于501 号池; 8 月25 日到9 月8 日, 501 号池氨氮、亚硝氮和硝氮浓度由高于303 号池转变为低于303 号池, 其对虾血清PO、SOD 活力由低于303 号池转变为高于303 号池。本实验中COD、BOD、pH 与盐度的耦合作用对斑节对虾血淋巴免疫指标的影响是明显的, 氨氮、亚硝氮和硝氮为影响斑节对虾血淋巴PO、SOD 等活力的关键因子。     

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.     

Immune responses and gene expression in hepatopancreas from Macrobrachium rosenbergii challenged by a novel pathogen spiroplasma MR-1008

Freshwater prawn Macrobrachium rosenbergii inoculated with 100 μl novel pathogen spiroplasma strain MR-1008 in logarithmic phase (10⁸ spiroplasmas ml^?1) were examined for alkaline phosphatase (AKP) activity, acid phosphatase (ACP) activity, superoxide dismutase (SOD) activity, catalase (CAT) activity, as well as expressions of 7 immune related genes in hepatopancreas after 1–28 d. Hematoxylin-eosin (HE) staining showed obvious pathological features in hepatopancreas connective and epithelial tissue. Enzyme activity analyze showed that hepatopancreas AKP and ACP activity increased markedly (P < 0.05) when inoculated with spiroplasma MR-1008 after 5 d and 10 d, respectively. SOD enzyme activity changed less obviously and slightly increased at 1 day post-inoculation, but CAT activity decreased significantly after 5 d inoculation. The expression levels of lipopolysaccharide and β-1,3-glucan-binding protein (LGBP), peroxinectin (PE), α2-macroglobulin (α2M), AKP, ACP, CAT, and copper/zinc SOD (Cu, Zn-SOD) genes in the hepatopancreas were examined by Real-Time PCR (qRT-PCR) and the results demonstrated that these immune related genes were induced by challenge with spiroplasma MR-1008. The results suggested that the prawn immune responses could be activated or inhibited by spiroplasma MR-1008, and that the hepatopancreas also plays key roles in innate immunity for defense against the pathogen.     

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。     

Effects of NaCl and Mycorrhizal Fungi on Antioxidative Enzymes in Soybean

The effects of different concentrations of NaCl on the activities of antioxidative enzymes in the shoots and roots of soybean (Glycine max [L.] Merr cv. Pershing) inoculated or not with an arbuscular mycorrhizal fungus, Glomus etunicatum Becker & Gerdemann, were studied. Furthermore, the effect of salt acclimated mycorrhizal fungi on the antioxidative enzymes in soybean plants grown under salt stress (100 mM NaCl) was investigated. Activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were increased in the shoots of both mycorrhizal (M) and nonmycorrhizal (NM) plants grown under NaCl salinity. Salinity increased SOD activity in the roots of M and NM plants, but had no effect on CAT and polyphenol oxidase activities in the roots. M plants had greater SOD, POD and ascorbate peroxidase activity under salinity. Under salt stress, soybean plants inoculated with salt pre-treated mycorrhizal fungi showed increased SOD and POD activity in shoots, relative to those inoculated with the non pre-treated fungi.     

刈割后黑麦草生理保护作用对其补偿性生长的影响

通过对黑麦草(Lolium perenne L)在轻度、中度、重度、全割刈割处理6 d和12 d后,残留叶片和叶片再生部分生长速率,超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活力,丙二醛、可溶性糖、脯氨酸含量的分析以揭示在刈割胁迫后叶片抗氧护酶活力和渗透调节物含量与其补偿性生长的关系,以及牧草耐刈性的生理调控机理。结果表明,轻度和中度及全割后叶片生长速率均高于对照,重度刈割低于对照。全割后叶片补偿性生长最明显、轻度和中度次之,重度刈割无补偿性生长。对照黑麦草叶片各部位抗氧化酶和渗透调节物含量不同,叶片顶部MDA含量较高,伴随着较高的SOD、CAT活力和较高的脯氨酸含量;叶片基部MDA含量最低,SOD、CAT活力及脯氨酸含量也较低。与对照相比,不同强度刈割6 d黑麦草再生叶和叶片平均MDA含量、SOD和CAT活力、可溶性糖和脯氨酸含量均较低。而不同强度刈割12 d,黑麦草再生叶和叶片平均MDA含量仍较低,但SOD和CAT活力增高,脯氨酸含量增加,POD活力和可溶性糖含量低于对照。这表明刈割在减少了叶面积,降低光合能力的同时,刈割伤害胁迫启动了牧草补偿性生长使残留叶片快速生长,而且残留叶片面积与其补偿生长速率成正相关。另外,虽然不同强度刈割下叶片补偿性生长速率不同,但不同强度刈割(12 d)均激活残留叶片抗氧化保护酶系统和促进脯氨酸积累。在补偿生长过程中,CAT和SOD能及时清除残留叶片中积累的氧自由基,维持较低的膜脂过氧化和细胞膜完整性,积累的脯氨酸能维护细胞水分平衡。因此,抗氧化酶(SOD和CAT)和渗透调节物(脯氨酸)在黑麦草刈割后受伤部位快速自愈及残留叶片快速补偿生长中起重要生理保护作用。     

Digestive enzymes present in crustacean feces as a tool for biochemical, physiological, and ecological studies

The effect of food composition on the digestive system of Penaeus vannamei shrimp was used to determine the suitability of feces for analysis of class, type, composition of digestive proteinases, and whether alterations in the digestive gland are mirrored in feces composition. Enzymes recovered from feces and the midgut gland of white shrimp P. vannamei were used for comparison purposes. Three groups of shrimp were assembled: two groups fed two different brands of commercial feeds (PI and SC) with different content of protein, and the last group fed 50% PI feed and 50% thawed giant squid. Composition of proteinases in the midgut gland and feces were identical, and trypsin and chymotrypsin paralogues were identified in both samples by substrate-electrophoresis. Total proteolytic, trypsin, and chymotrypsin enzyme activities were higher in both samples from organisms fed SC, than in the other two groups. In the hepatopancreas, trypsin activity was ∼30% higher in SC fed group. Final average weights of shrimp were close in three groups, but hepatopancreas weight was 20% higher in the SC group. The degree of protein hydrolysis (DH) in vitro for the SC and PI was evaluated by the pH-stat method, using enzymes from feces and hepatopancreas of each group. The DH of food was no different, but it was affected by enzyme source, hepatopancreas extract (HPE) or feces extract (FE). DH was always higher when FE was the enzyme source than when HPE was the source. The proposed methods for recovery of enzymes from shrimp feces can be applied to other crustaceans. Measurements were sufficiently sensitive to allow quantifying the effects of feed on digestion physiology and other ecological and physiological applications, without the necessity of killing specimens.     

Effects of simulated acid rain on the physiology of carmine spider mite, Tetranychus cinnabarinus (Boisduvals) (Acari: Tetranychidae)

Abstract:  The physiological effect of simulated acid rain sprayed on carmine spider mite Tetranychus cinnabarinus (Boisduvals) and host plant, were measured in a series of laboratory trials. We examined potential changes in three kinds of protective enzymes [peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT)] and three hydrolases [acid phosphatase (ACP), alkaline phosphatase (ALP) and carboxylesterase (CarE)] in response to changes in pH values of simulated acid rain at different time of exposure. POD, SOD and CAT activities increased significantly with the increase in the acidity of the acid rain, reaching the highest levels at pH 4.0 or 3.0, and then declined. Changes in ACP activity were similar to those observed in the protective enzymes. The increasing extent of the activities of these four enzymes after 30 and 45 days treatment became smaller than that after 15 days treatment . ALP activities decreased as pH value declined. There were no significant changes in CarE activities after 15 and 45 days, but that in pH 4.0 and 3.0 decreased after 30 days. The enhanced anti-oxidation enzyme levels (POD, SOD and CAT) and ACP activities in pH 4.0 and 3.0 reduced the effects of these toxic products on mites, resulting in the strengthening of the defensive power, and increase in survival and reproductive power of the mites, thus leading to an increase in the density of mites on host plant. From these results, we inferred that POD, SOD, CAT and ACP might be relevant to population changes of mites under acid rain pressure.