Effects of hypoxia on immune responses and carbohydrate metabolism in the Chinese mitten crab,Eriocheir sinensis

In aquaculture, hypoxia is an important stressor that can inhibit the healthy development of Chinese mitten crab, Eriocheir sinensis. The aim of this work was to research on how hypoxia (1 mg/L) followed by reoxygenation affects the immune response and carbohydrate metabolism in the Chinese mitten crab. The activities of immune‐related enzymes including lysozyme (LZM), acid phosphatase (ACP), alkaline phosphatase (AKP), superoxide dismutase (SOD) and phenoloxidase (PO) and concentration changes in haemolymph glucose, trehalose and hepatic glycogen were evaluated after 8 hr of hypoxia followed by 12 hr of reoxygenation. After 8 hr of hypoxia, the activities of LZM, PO, SOD and hepatic glycogen concentration significantly decreased to the minimum when compared to the control group, but the activities of ACP and AKP, and haemolymph glucose concentration significantly increased. After 12 hr of reoxygenation, the activities of LZM, ACP, AKP and SOD recovered to the normal level except SOD in plasma, while the PO, SOD in plasma and hepatic glycogen concentration were still significantly lower than the control level. No significant change was observed in trehalose concentration during hypoxia and reoxygenation periods. These results indicated that although the activities of ACP and AKP were likely increased to facilitate the hydrolysis of foreign matter, the overall strength of the immune response was decreased as LZM, PO and SOD activities were suppressed after exposure to hypoxic conditions. The energy stores of exposed crabs were reduced due to the expenditure of energy during exposure to hypoxic stress. In general, chronic hypoxia would impair the immunocompetence and increase the energy expenditure of Chinese mitten crab, further increased the risk of pathogen infection and feed coefficient, so the negative effects of hypoxia should not be ignored.     

三疣梭子蟹血淋巴免疫功能的初步研究

利用API ZYM试剂盒和酶活力测试盒检测了三疣梭子蟹血细胞及血清中的19种酶类及血清中溶菌酶、碱性磷酸酶、酸性磷酸酶、超氧化物歧化酶和过氧化物酶的活性。试验结果表明,在雄蟹血清和血细胞中均检测到19种酶,在雌蟹血清和血细胞中分别检测到18种和16种。碱性磷酸酶和酸性磷酸酶的活性很低。在此基础上,研究了三疣梭子蟹血清和血细胞溶解物(HLS)对溶藻弧菌的抗菌活力,结果显示血清样品对溶藻弧菌的抗菌活力较强,但其体外作用的持续时间较短;而血细胞溶菌酶对溶藻弧菌的抗菌活力较弱。     

Effect of dietary taurine supplementation on growth,digestive enzyme,immunity and resistant to dry stress of rice field eel (Monopterus albus) fed low fish meal diets

This study was conducted to evaluate the effect of dietary taurine supplementation on growth, immunity and resistant to dry stress of rice field eel (Monopterus albus) fed low fish meal diets. Six isonitrogenous and isolipid diets (32% fish meal) supplemented with six taurine concentrations (0, 0.3, 0.6, 0.9, 1.2 and 1.5 g/kg; designated as T₀, T_0.03, T_0.06, T_0.09, T_0.12 and T_0.15 groups, respectively) were prepared. A diet including 42% fish meal (FM group) was also included as a reference. The results showed that specific growth rate (SGR) in FM group was significantly higher than that in lower fish meal treatments. SGR significantly increased and slowly decreased with the increase in taurine supplementation level. Lipase activity value in intestine of M. albus fed FM diet was maximum, and with the increase in taurine supplementation level, lipase activity significantly increased and slowly decreased. The FM group had relative higher total antioxidant capacity (T‐AOC) content, catalase (CAT), total superoxide dismutase (T‐SOD), and lyzozyme (LZM) activities in serum than the other groups. With the increase in dietary taurine supplementation level, the CAT, T‐SOD, T‐AOC and LZM activities in serum significantly increased and then decreased. In the dry stress experience, the adrenaline (AD), cortisol (COR), glucose (GLU), total cholesterol (CHOL), and malondialdehyde (MDA) concentrations, T‐AOC content, CAT and T‐SOD activities in serum of M. albus in the four groups first increased and reached the peak at 2 hr, and then decreased under air‐exposure stress. Compared to the FM group, T_0.15 group had relative higher T‐AOC content, CAT and T‐SOD activities, and lower AD, COR GLU, TC and MDA concentrations.     

氨氮急性胁迫及其毒后恢复对红螯光壳螯虾幼虾相关免疫和代谢指标的影响

应用实时荧光定量PCR技术,结合生物酶和代谢产物测定,研究了氨氮急性胁迫对红螯光壳螯虾幼虾代谢及免疫系统的毒性影响及其毒后恢复情况。实验首先进行3 d的氨氮胁迫,取样后剩余虾移入曝气自来水进行7 d的毒后恢复实验。结果表明,3 d氨氮胁迫后,肌肉ACP、AKP、SOD活性表达均受到显著影响,随着氨氮浓度的升高酶活性分别降低,最高浓度组(16 mg/L)降低为对照组的76%、68%和62%。线粒体MnSOD、胞外Cu/ZnSOD的mRNA表达量也随着氨氮浓度增加而下降,最高浓度组降低至对照组的69%和68%。CAT、GPX活性以及GPX和GST的mRNA表达量变化不显著。肝胰腺中可溶性蛋白和甘油三酯含量随着氨氮浓度升高而降低,最高浓度组分别降低至对照组的72%和59%,AST活性在12 mg/L浓度组显著升高至对照组的134%。7 d恢复期过后,ACP和AKP活性以及各代谢指标恢复到正常水平;而SOD和GPX活性高于对照组。各抗氧化基因的表达量都不同程度高于对照组。实验表明,高浓度氨氮胁迫能抑制部分免疫相关酶的活性及基因表达,对免疫系统造成损害。氨氮胁迫下,红螯光壳螯虾动员蛋白质和脂肪来供能应对胁迫。7 d的恢复时间不足以让红螯光壳螯虾从胁迫中完全恢复,其肌肉仍处于轻度氧化应激状态。     

不同浓度诺氟沙星对中国对虾非特异性免疫酶活的影响

向对虾配合饲料中添加不同剂量的诺氟沙星投喂中国对虾7d,分析不同时间诺氟沙星对中国对虾肌肉及鳃溶菌酶(LSZ)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、碱性磷酸酶(AKP)及酸性磷酸酶(ACP)活性的影响.结果表明,15mg/kg诺氟沙星可以显著抑制中国对虾肌肉SOD活力(P＜0.05),而对鳃SOD活力则整体呈现促进作用,该浓度组肌肉和鳃CAT及LSZ活力整体高于对照组,而AKP和ACP活力则显著低于对照组(P＜0.05);30 mg/kg诺氟沙星对中国对虾肌肉SOD活力呈现先抑制后促进的作用,而对鳃SOD活力则整体呈现抑制作用,该组肌肉和鳃CAT活力显著高于对照组(P＜0.05),肌肉LSZ活力呈现先下降后上升的趋势,而鳃LSZ活力则与对照组没有显著差异;向饲料中添加60 mg/kg诺氟沙星对中国对虾肌肉、鳃SOD、CAT、LSZ活力整体呈现促进作用,而对AKP和ACP活性则呈现显著抑制作用(P＜0.05).     

Hypoxia‐induced changes in survival,immune response and antioxidant status of the Pacific white shrimp (Litopenaeus vannamei) fed with graded levels of dietary myo‐inositol

A 3‐hr experiment was conducted to investigate the effects of dietary myo‐inositol (MI) supplementation on survival, immune response and antioxidant abilities in Litopenaeus vannamei under acute hypoxia stress. Six practical diets were formulated with supplementation of graded levels (control group 0, 0.1, 0.2, 0.4, 0.8 and 1.6 g/kg dry diet) of MI and were randomly assigned to triplicate groups of L. vannamei (mean weight 0.40 ± 0.00 g) for 8 weeks. Ten healthy shrimp (final mean weight approximately 11–14 g) randomly selected from each tank were exposed to hypoxia stress after feeding trial. After 3‐hr acute hypoxia stress, survival of shrimp fed MI‐supplemented diets (except 0.1 and 0.4 g/kg diets) was significantly increased compared with the control group. Shrimp fed control diet had lower activities of alkaline phosphatase (AKP), acid phosphatase (ACP), total antioxidant capacity (T‐AOC) and glutathione peroxidase (GPX), and higher malondialdehyde (MDA) and protein carbonyl (PC) contents in hepatopancreas than those fed the MI‐supplemented diets. In addition, mRNA expression levels of heat shock protein 70 (Hsp70), catalase (CAT) and penaeidin were significantly differentially regulated in hepatopancreas. In summary, dietary MI supplementation may have a positive effect on improving resistance to acute hypoxia stress of L. vannamei.     

副溶血弧菌感染日本蟳后对几种免疫活性酶的影响

用副溶血弧菌悬液和生理盐水(对照)注射感染日本蟳,研究其血清超氧化物歧化酶(SOD)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、过氧化氢酶(CAT)活性的变化,并进行统计学分析。试验结果表明,注射副溶血性弧菌悬液组12、24、48 h后SOD活性均高于对照组,48 h达到最高;122、4 h CAT活性增加明显,24 h达最高;AKP活性5、12、24 h均有增加,高于对照组,24 h最高;ACP活性12 h开始上升,24 h达最高。在人工感染副溶血性弧菌48 h内,日本蟳体内的免疫因子发生明显的变化,血清中SOD、ACP、AKP、CAT活性在不同时间段均有显著升高趋势,高于对照组。     

复方中草药对杂色鲍幼鲍血淋巴中几种酶活力的影响

将12种中草药按其不同的药理作用和药效进行配伍,形成5组复方中草药,分别编号为中草药1～5号。将以上中草药添加到杂色鲍幼鲍的人工配合饲料中,连续喂饲幼鲍,饲养47d后,测定幼鲍无细胞血淋巴中超氧化物歧化酶(SOD)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、溶茵酶(LSZ)活力。结果显示,所有添加中草药的sOD活力均比对照组降低,中草药1、5号可提高幼鲍血淋巴的ACP和LSZ活力,而对AKP则起到降低的作用;中草药3、4号可提高幼鲍血淋巴的LSZ活力,却降低幼鲍血淋巴的ACP和AKP活力,中草药2号对ACP、AKP和LSZ活力均有提高作用,具有较突出的促进杂色鲍幼鲍免疫酶活力的功能。     

盐度胁迫对三疣梭子蟹血清非特异性免疫因子的影响

采用紫外分光光度法和酶学分析方法研究了盐度(14和36)胁迫下12 h内经溶藻弧菌筛选过的三疣梭子蟹F2代(简称筛选蟹F2)的血蓝蛋白含量及血清酶活力的变化,用普通三疣梭子蟹F1代(简称普通蟹F1)作对照。试验结果表明,在高盐和低盐短时胁迫下,筛选蟹F2和普通蟹F1的血蓝蛋白含量、3种保护酶(SOD、POD和CAT)和磷酸酶(ACP和AKP)活力随时间变化呈现有升有降的变化趋势。此外,无论在低盐还是高盐胁迫下,筛选蟹F2的血蓝蛋白含量和各项血清酶活力分别在0 h(正常水平)和盐度胁迫下均显著高于普通蟹F1(P<0.05)。低盐和高盐短时胁迫对三疣梭子蟹的非特异性免疫影响显著且表现出较强的应激反应;在正常和抗盐度胁迫下,筛选蟹F2的非特异性免疫比普通蟹F1更具优势。     

Effect of dietary β‐1,3‐glucan on the immune response of Litopenaeus vannamei exposed to nitrite‐N

β‐1,3‐Glucan with 1.0 g kg^?1 was supplemented to a basal diet [control (C)] with different feeding schedules: permanently β‐1,3‐glucan diet (BG) and 14 days BG–14 days control diet (C + BG). Growth and immunological responses [respiratory burst (RB), superoxide dismutase (SOD), catalase (CAT), lysozyme and total protein] in haemolymph, hepatopancreas and muscle of Litopenaeus vannamei were recorded after 84‐day feeding and exposure to nitrite‐N (20 mg L^?1) for 120 h. β‐1,3‐Glucan administration did not affect growth performance. However, as compared with control, elevated CAT and lysozyme activities were seen in haemolymph of both BG groups, while significantly higher activities of SOD, lysozyme and RB in hepatopancreas, and higher activities of CAT and lysozyme in muscle were only seen in C + BG group. After nitrite‐N stress, significantly higher haemolymph protein, and hepatopancreas activities of lysozyme and RB were observed in both BG groups, but significantly higher activities of haemolymph SOD was only seen in C + BG group. The mortality in groups BG and C + BG was significantly lower than that in group C, but C + BG group showed a trend of higher nitrite‐N resistance compared with BG group. Considering dietary cost and immunostimulatory effects, the feeding schedule with 14 days BG–14 days control diet is more recommended.     

低氧胁迫对西伯利亚鲟幼鱼生理状态的影响

为探究低氧胁迫对西伯利亚鲟幼鱼的血液基础指标、抗氧化和能量代谢相关酶活力的影响,以体质量为(19.46±4.9)g的西伯利亚鲟幼鱼为研究对象,开展了低氧(2.3mg/L)胁迫3h和自高氧(18.5mg/L)逐渐降至低氧(2.8mg/L)又恢复到正常溶氧(7mg/L),这两种不同情况下的低氧胁迫试验。检测了血液红细胞数、血红蛋白质量浓度,以及肝脏和肌肉的抗氧化酶活力和鳃组织三磷酸腺苷酶活力。结果显示,低氧胁迫下西伯利亚鲟幼鱼血红细胞数升高,血红蛋白质量浓度、血液pH、肝脏和肌肉的超氧化物歧化酶和过氧化氢酶活力以及碱性磷酸酶活力均低于正常溶氧组,而鳃组织钠/钾—三磷酸腺苷酶活力无明显变化。试验结果显示,急性低氧胁迫能较快使西伯利亚鲟幼鱼血液红细胞数目增加,并影响肝脏、肌肉的抗氧化应激及代谢相关酶活力,但是短时间内西伯利亚鲟幼鱼可能还未实现鳃部的能量调节。     

三疣梭子蟹患"乳化病"后几种保护酶活力的变化

以酶活力试剂盒测定法研究三疣梭子蟹患“乳化病”后主要病变组织中保护酶系统的病理变化。以患病三疣梭子蟹的肝胰腺、鳃、心肌及步足肌组织为材料，研究4种组织中淀粉酶（AMS）、超氧化物岐化酶（SOD）、溶菌酶（Lysozyme）、过氧化氢酶（CAT）、碱性磷酸酶（AKP）、酸性磷酸酶（ACP）、谷草转氨酶（AST／GOT）和谷丙转氨酶（ALT／GPT）8种酶活力及丙二醛（MDA）指标的变化。试验结果表明：病变后梭子蟹4种组织中9种指标均存在不同程度的病理变化，表明酶活力指标可以作为乳化病病理分析的重要参考依据。     

The in vitro effects of divalent metal ions on the activities of immune‐related enzymes in coelomic fluid from the sea cucumber Apostichopus japonicus

The immune‐related enzymes in marine animals are very sensitive to divalent metal ions. To investigate the roles divalent metal ions play in the influence on the immunity of sea cucumber Apostichopus japonicus, one of the most important commercial species in Asian countries, the effects of eight divalent metal ions at concentrations of 2.5, 5, 10, 15, 20, 25 and 30 mmol L^?1 on the activities of superoxide dismutase (SOD), phenoloxidase (PO), acid phosphatase (ACP), alkaline phosphatase (AKP) and myeloperoxidase (MPO) in coelomic fluid were determined with the nitro blue tetrazolium chloride (NBT) method, dopachrome formation method, p‐nitrophenyl phosphate (pNPP) method and 3,3′,5,5′‐tetramethyl benzidine (TMB) method. The results indicated that Mg²⁺ enhanced the activities of SOD, PO, ACP and AKP significantly and showed no obvious effect on MPO activity; Zn²⁺ increased the activities of SOD, ACP and AKP, and showed no obvious effect on the activities of PO and MPO; Cu²⁺ enhanced the activities of ACP, AKP and MPO and activated SOD and PO at a certain concentration range; Ca²⁺ and Mn²⁺ inhibited the activities of ACP and AKP; Fe²⁺ had strong inhibitory effect on SOD activity; Pb²⁺ showed inhibitions on the activities of SOD, PO, ACP and AKP; and Cd²⁺ inhibited MPO activity greatly. The data obtained in this study collectively suggest that Mg²⁺, Zn²⁺ and Cu²⁺ have potential in promotion of A. japonicus immunity, while Ca²⁺, Fe²⁺, Pb²⁺, Cd²⁺ and Mn²⁺ might be limiting factors to the immune response of A. japonicus.     

Effects of β‐conglycinin on growth performance,antioxidant capacity and intestinal health in juvenile golden crucian carp,Carassius auratus

The objective of this study was conducted to research the effects of β‐conglycinin in the diets on the growth performance, immunity function, antioxidant capacity and intestinal health of juvenile golden crucian carp (Carassius auratus). Five diets contained respectively (0, 20, 40, 60 and 80 g/kg) β‐conglycinin, and were used to feed juvenile golden crucian carp for 56 days. Final weight, weight gain and specific growth rate were significantly reduced by dietary β‐conglycinin (20–80 g/kg). Feed efficiency and protein efficiency were significantly reduced by dietary β‐conglycinin (40–80 g/kg). In hepatopancreas, the activities of T‐SOD, ACP, ALT and T‐AOC were significantly suppressed by dietary β‐conglycinin (20–80 g/kg). The activities of LZM, AKP, CAT and GPx were significantly reduced by dietary β‐conglycinin (40–80 g/kg). The activities of protease were significantly reduced and the content of MDA was significantly increased by dietary β‐conglycinin (60–80 g/kg). In proximal intestines, the activities of protease and CAT were significantly decreased by dietary β‐conglycinin (40–80 g/kg). In mid and distal intestines, the activities of protease and CAT were significantly inhibited by dietary β‐conglycinin (20–80 g/kg). In intestines, T‐AOC and GPx were significantly declined by dietary β‐conglycinin (20–80 g/kg). In proximal and mid intestines, the content of MDA were significantly increased by dietary β‐conglycinin (40–80 g/kg). In distal intestines, the content of MDA was significantly increased by dietary β‐conglycinin (20–80 g/kg). The expression of IGF‐I was significantly decreased and the expression of IL‐1β and TNF‐α was significantly increased by dietary β‐conglycinin (20–80 g/kg). The structural integrity of intestinal tissues were damaged by dietary β‐conglycinin (20–80 g/kg), the part of intestinal villus were shed, the part of epithelial cells were separated from lamina propria. Ultimately, these results suggested dietary β‐conglycinin should be <20 g/kg in formula feed of golden crucian carp.     

中华绒螯蟹感染“颤抖病”病毒后血清免疫因子的变化

对中华绒螯蟹进行颤抖病病毒悬液和PBS(对照)注射感染,研究其血清免疫因子抗菌活性(Ua)、溶菌活性(UL)、凝集活性(HA)、酚氧化酶(PO)、过氧化酶(POD)、超氧化物歧化酶(S0D)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)活性的变化,并对实验的结果进行统计学分析。结果表明,在中华绒螯蟹人工感染颤抖病病毒后,蟹体内的免疫因子发生了明显的变化,血清中Ua、UL活性先是降低,然后上升至高于正常值水平。P0活性先上升然后下降至低于正常值水平,POD、HA、SOD活性逐渐升高至显著水平;ACP及AKP活性则显著下降。     

鳗弧菌感染对日本鑚部分免疫活性因子的影响

对日本蟳(Charybdis japonica)进行鳗弧菌(Vibrio anguillarum)悬液和生理盐水(对照)注射感染,研究其血清免疫因子超氧化物歧化酶(SOD)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、过氧化氢酶(CAT)活性的变化,并对试验结果进行统计学分析。结果表明,注射鳗弧菌悬液组SOD活性12、24、48h均高于对照组,48h达到最高;CAT活性12、24h增加非常明显,12h达最高;AKP活性5、12、24h都有增加,均高于对照,24h最高;ACP活性12h开始上升,24h达最高。在人工感染鳗弧菌48h内,日本蟳体内的免疫因子发生了明显的变化,血清中SOD、ACP、AKP、CAT活性在不同时间段均有显著升高趋势,高于对照组水平。     

温度胁迫对大鳞副泥鳅抗氧化与非特异性免疫指标的影响

为研究大鳞副泥鳅(Paramisgurnus dabryanus)对温度胁迫的适应能力,开展了温度胁迫与恢复实验。设置对照组(22℃)、高温组(29℃)、降温组(15℃)与低温组(8℃)4个处理,在胁迫期的第1、2、4、8、12h与恢复期第4、8、12、24h取肝脏,测定其超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、溶菌酶(LZM)、碱性磷酸酶(AKP)活力和丙二醛(MDA)、血清谷丙转氨酶(ALT)、谷草转氨酶(AST)含量。结果显示,胁迫期内肝脏SOD和CAT活力均在温度骤变的影响下受到抑制,显著低于对照组水平(P0.05),MDA含量则显著升高(P0.05);恢复期内各实验组SOD和CAT活力均逐渐升高,MDA含量逐渐降低,至恢复期结束时,对照组与各实验组间无显著差异(P0.05)。温度骤变条件下,各实验组肝脏LZM与AKP活力显著降低(P0.05),恢复期内高温组LZM与AKP活力无明显变化,降温组与低温组LZM与AKP活力则逐渐升高,至恢复期结束时与对照组无显著差异(P0.05)。温度胁迫显著升高血清中AST与ALT含量(P0.05),恢复期内各实验组AST与ALT含量均呈下降趋势,至恢复期结束时,除高温组ALT含量外均降至对照组水平。研究表明,温度胁迫显著降低大鳞副泥鳅SOD、CAT、LZM与AKP活力,MDA与血清转氨酶AST、ALT含量升高,其中,高温胁迫会对大鳞副泥鳅非特异性免疫机能造成不可逆破坏,其他抑制均为可恢复的。     

Analysis of Streptococcus agalactiae‐induced liver injury in tilapia (Oreochromis niloticus)

Streptococcosis, a bacterial disease for multiple fish species, especially tilapia, caused more disruption to the fish culture industry. However, the underlying mechanisms for the occurrence of streptococcosis remain unclear. The aim of this study was to study the effects of streptococcus on the liver of tilapia. In this study, tilapia were injected streptococcal solution (0.05 ml/10 g body weight) and collected blood and liver at 4, 8, 24, 48, 72, 96, 120 and 144 hr after injection. The results showed that the activities of glutamate oxalate transaminase (GOT), glutamate pyruvate transaminase (GPT), alkaline phosphatase (AKP) and superoxide dismutase (SOD) in the serum of tilapia exhibited irregular changes: increase at first, then decrease and increase again. The levels of total antioxidant capacity (T‐AOC), glutathione (GSH) and malondialdehyde (MDA) showed a decreasing tendency in serum after injection, except GSH at 4 hr. The activity of catalase (CAT) decreased in serum at 4, 24, 120 and 144 hr after injection of streptococcal solution, while the content TP enhanced in serum at 48, 72 and 120 hr after injection. Observations of pathological sections showed obvious damage to the liver tissue structure in response to streptococcal infection. Western blotting revealed an increase in ikkbeta (Ikkβ) protein with prolonged infection time and a decrease in NF‐kappa‐B inhibitor alpha (IkB‐α) and ikappab kinase alpha (Ikkα) proteins. The gene expression of cytochrome P4501A (CYP1A) was downregulated in liver at 24 and 48 after injection, while the mRNA levels of heat stress protein 70 (HSP70) and tumour necrosis factor alpha (TNF‐α) increased at 48 and/or 72 hr. Taken together, our findings confirmed that streptococcus can cause serious damage to the liver of tilapia.