仿刺参酸性磷酸酶的提取及粗酶性质研究

试验结果表明,仿刺参酸性磷酸酶的最佳提取缓冲液为 pH 7.5的Tris-HCl缓冲液,最佳沉淀条件为饱和度为80 %的硫酸铵.仿刺参体壁和肠道中的酸性磷酸酶最适反应条件均为:40 ℃,pH 4.0.Ca2+、Mg2+和Mn2+对体壁酸性磷酸酶有激活作用,而Fe3+、Fe2+、Zn2+、Cu2+、Ba2+对其有抑制作用,其中Cu2+的抑制作用最强;肠道酸性磷酸酶表现的特征与之类似,但Zn2+的抑制作用最强.PAGE电泳结果表明,仿刺参体壁和肠道中的酸性磷酸酶类型不同,肠道中有3种同工酶.     

仿刺参补体B因子溶血活性初步研究

本文应用高等哺乳动物血清补体B因子溶血检测技术检测仿刺参个体体内补体B因子。通过溶血法检测了健康仿刺参以及人工诱导化皮仿刺参体腔液补体B因子活性。结果显示仿刺参体腔液有补体B因子存在,且含补体B因子仿刺参体腔液溶血活性明显高于去补体B因子体腔液的溶血活性。实验对仿刺参体腔液进行加热处理结果显示其补体B因子是对热不稳定的分子,此特性与高等哺乳动物B因子特征相同。仿刺参体内存在补体B因子,它与脊椎动物补体B因子性质相似,在免疫旁路途径中起重要作用。     

仿刺参补体B因子溶血活性初步研究

应用高等哺乳动物血清补体B因子溶血检测技术，检测了健康仿刺参以及人工诱导化皮仿刺参体腔液补体B因子活性。结果显示仿剌参体腔液有补体B因子存在，且含补体B因子仿刺参体腔液溶血活性明显高于去补体B因子体腔液的溶血活性。实验对仿刺参体腔液进行加热处理，结果显示其补体B因子是对热不稳定的分子，此特性与高等哺乳动物B因子特征相同。仿刺参体内存在补体B因子，它与脊椎动物补体B因子性质相似，在免疫中起重要作用。     

低盐急性胁迫对仿刺参相关生理指标的影响

通过模拟仿刺参养殖池塘雨季的盐度变化,选用2龄仿刺参(Apostichopus japonicus)为研究对象,刺参体重(16.93±3.08)g,研究盐度骤降及胁迫恢复对仿刺参体腔液相关生理指标的影响。盐度先由30以每6 h变化3个盐度的速度下降至18,然后在盐度18保持96 h,随后盐度以相同速度上升恢复至30,并保持24 h。结果显示,各盐度取样点间仿刺参体腔液渗透压、体腔液总蛋白浓度、Na+、K+、Cl–浓度与盐度的变化趋势一致,Na+、K+、Cl–浓度均在盐度下降到18时达到最低值,分别为(131.15±14.42)mmol/L,(6.08±0.24)mmol/L和(141.76±2.13)mmol/L;而Ca2+浓度一直呈上升趋势。体腔液中Na+-K+-ATP酶活力在盐度18并保持4 d后显著高于其他组(P0.05),盐度对其体腔液中Na+-K+-ATP酶活力的影响与体腔液渗透压变化趋势不一致。盐度胁迫对谷丙转氨酶活力无显著影响。结果表明,盐度胁迫对仿刺参渗透调节能力有显著影响,实验中体腔液渗透压与体腔液Na+、K+、Cl–浓度有一定的相关性。盐度胁迫对仿刺参的呼吸代谢产生的影响不显著。研究结果为丰富仿刺参适应环境盐度的机制提供基础资料,为进一步了解盐度胁迫下刺参的生理生态学特征以及今后的刺参增养殖生产提供参考。     

尼罗罗非鱼淀粉酶性质的初步研究

研究了尼罗罗非鱼(Oreochromis niloticus)肝脏淀粉酶的基本性质及金属离子对该鱼淀粉酶活性的影响。结果表明:罗非鱼淀粉酶活力的最适pH是6.5,最适底物浓度是2%。研究金属离子对该鱼淀粉酶活力影响,一价金属离子K+、Li+、Na+对酶活力影响较小;二价金属离子Cu2+对酶活力具有抑制作用,Zn2+对酶活力无明显影响;三价金属离子Al3+对酶活力具有抑制作用,但效果不是很强烈,Fe3+对酶活力具有明显的激活作用;碱土金属Mg2+、Ca2+对淀粉酶活性有激活作用,而Ba2+对酶具有抑制作用;重金属离子Cd2+、Pb2+有明显的抑制作用。     

低盐胁迫对刺参非特异性免疫酶活性及抗菌活力的影响

以养殖刺参为研究对象,测定了不同强度盐度胁迫对刺参生存、生长及体腔液中超氧化物歧化酶(SOD)、溶菌酶（LZM）活性及抗菌活力的影响。结果表明,1）盐度16实验组刺参存活率最低,为66.7%,与其他各实验组及对照组差异显著（P<0.05）;盐度18、20实验组存活率分别为88.9%、92.6%,无显著差异（P＞0.05）;盐度≥22时,刺参能够全部存活,实验组刺参生长与对照组相比差异显著（P<0.05）。盐度≤20时,刺参生长表现为负增长。2）刺参体腔液中SOD、LZM活性及抗菌活力随时间变化呈现总体降低的趋势。实验开始时,各实验组SOD、LZM活性及抗菌活力均为最高值,30 d时,各实验组SOD、LZM活性及抗菌活力降至最低。低盐胁迫会显著影响刺参正常生存、生长,并导致刺参免疫力降低,增加对病原菌的易感性,从而诱导疾病发生并造成死亡。     

饲料中间隔添加壳寡糖对仿刺参生长、非特异性免疫和抗病力的影响

壳寡糖是良好的免疫增强剂,但在水生动物的应用还十分有限。本研究通过间隔投喂方式研究壳寡糖对仿刺参生长性能、非特异性免疫能力、消化能力、组织学和抗病力的影响。实验挑选体质量(18.51 ± 0.28)g仿刺参,对照组饲喂基础饲料,实验组以3天1次的饲喂频率饲喂含0.5%的壳寡糖饲料,其余时间饲喂基础饲料,进行8周的养殖试验后,测定该饲喂方式下仿刺参的生长性能、非特异性免疫能力、肠道消化酶、肠道和呼吸树组织学以及对灿烂弧菌的抗病力。结果显示,3天1次的饲喂频率下,仿刺参特定生长率和存活率几乎未受影响,但显著提高了脏壁比和肠壁比(P < 0.05);在免疫指标方面,显著提高了仿刺参体腔细胞的吞噬活性和呼吸爆发能力(P < 0.05),以及体腔细胞和肠道的酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、溶菌酶(LZM)和总一氧化氮合酶(T-NOS)的活性(P < 0.05),其中肠道AKP和LZM活性分别提高了70.06%和156%,且acp、akp、lzm基因表达量显著提高(P < 0.05),其中肠道akp和lzm基因表达量分别提高了13.47%和22.36%;抗氧化指标结果显示,显著提高了仿刺参体腔细胞过氧化氢酶(CAT)活性(P < 0.05),而体腔细胞和肠道的丙二醛(MDA)含量无显著差异(P > 0.05);组织学结果显示,该饲喂频率显著提高了前肠肌肉层厚度和浆膜层厚度,中肠和后肠的皱襞高度和宽度;灿烂弧菌(Vibrio splendidus)攻毒结果显示,实验组仿刺参的相对保护率达到66.67%。综上,3天1次的饲喂频率能够一定程度上提高仿刺参的生长性能,非特异性免疫酶活性,明显改善肠道结构,研究结果可为壳寡糖对仿刺参作用机制的研究及投喂频率的确定提供数据支撑和理论依据。     

仿刺参β-胸腺素基因的克隆和表达分析

β-胸腺素是一种具有抗菌活性的多肽。采用RACE方法克隆仿刺参β-胸腺素(β-Thymosin),命名为Ajβ-Thymosin。Ajβ-Thymosin基因的序列全长1877 bp,开放阅读框为126 bp,编码41个氨基酸;分子质量为4.6 ku,理论等电点为5.25。仿刺参β-胸腺素中存在着保守的功能序列EVASFD-KSKLK和保守的G-actin结合结构域LKKTET。系统发育进化分析结果显示,仿刺参β-胸腺素和紫色球海胆的β-胸腺素聚为一支。采用实时荧光定量PCR检测Ajβ-Thymosin基因在仿刺参不同组织和不同发育时期的表达量,发现其在体腔细胞中的表达量最高,在小耳幼虫期开始高表达。经过假交替单胞菌、灿烂弧菌、蜡样芽孢杆菌和希瓦氏菌4种病原菌刺激后,仿刺参体腔细胞中Ajβ-Thymosin基因的表达量在4 h均受到抑制;在蜡样芽孢杆菌和希瓦氏菌组,Ajβ-Thymosin基因的表达量在12 h开始上调;在假交替单胞菌组,Ajβ-Thymosin基因的表达量在24 h开始上调;在灿烂弧菌组,Ajβ-Thymosin基因的表达量在48 h才开始上调。病原刺激产生的动态表达...     

全营养破壁酵母对仿刺参非特异性免疫及肠道菌群的影响

仿刺参(Apostichopus japonicus)体质量(0.65±0.03)g,基础饲料中添加5%(质量分数)全营养破壁酵母为实验饲料,以基础饲料为对照。分别于投喂后第15天和第30天检测仿刺参体腔细胞数量、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、过氧化氢酶(CAT)活性,于第30天计数肠道菌群。结果显示,实验组仿刺参体腔细胞数量与对照组差异不显著;ACP、AKP、CAT活性均比对照组有所增高,其中第15天CAT活性和第30天AKP活性与对照组具有显著性差异(P<0.05)。投喂破壁酵母对仿刺参肠道菌群有一定影响,可显著降低肠道内异养菌总数及弧菌数量。研究表明,全营养破壁酵母对仿刺参非特异性免疫活性有增强作用,且对肠道有害菌群有一定抑制作用,但其对肠道菌群的影响及适宜添加量等需进一步研究。本研究旨在探索全营养破壁酵母在海参育保苗中的作用机理,以期为仿刺参安全健康养殖开发新型绿色添加剂提供科学依据。     

一株芽孢杆菌的分离、鉴定及其对仿刺参免疫的影响

从大连湾地区健康的刺参养殖池塘沉积物中分离出一株优势菌株B4,通过菌落特征、生理生化特性和16SrDNA同源性分析,确定菌株B4为芽孢杆菌属。该菌株在温度20~35℃,pH 6~9,盐度3~7.5下生长良好。采用108cfu/mL密度的B4菌液投喂仿刺参,投喂后第0、7、14、21d和第28d测定仿刺参体腔液中酸性磷酸酶、碱性磷酸酶、超氧化物歧化酶、溶菌酶和酚氧化酶的活性。试验结果表明,第7d时,试验组仿刺参腔液中碱性磷酸酶、超氧化物歧化酶和酚氧化酶的活性显著高于对照组(P0.05);第21d时,试验组仿刺参腔液中溶菌酶的活性显著高于对照组;第28d时,酸性磷酸酶的活性与对照组具有极显著差异(P0.01),说明B4可提高仿刺参体腔液中酸性磷酸酶、碱性磷酸酶、超氧化物歧化酶、溶菌酶和酚氧化酶5种免疫酶的活性,对仿刺参有一定的免疫刺激作用。     

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 Na/K and Mg/Ca ratios in saline groundwaters on Na-K-ATPase activity, survival and growth of Marsupenaeus japonicus postlarvae

The effects of the Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios in saline groundwaters on Na⁺-K⁺-ATPase activity, survival and growth of Marsupenaeus japonicus postlarvae were investigated. The results indicate that the Na⁺-K⁺-ATPase activity, survival rate and weight gain of postlarvae were significantly affected by the Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios (P < 0.05). The Na⁺-K⁺-ATPase activity of postlarvae, in every treatment, changed corresponding to Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios, and came to a stable level after 24 h. There was a negative relation between Na⁺-K⁺-ATPase activity and Na⁺/K⁺ ratio, while there was a positive relation between Na⁺-K⁺-ATPase activity and Mg²⁺/Ca²⁺ ratio. Compared with seawater (the Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios are 27.8 and 4.64 respectively), the Na⁺-K⁺-ATPase activity of the Na⁺/K⁺ ratio 30 treatment showed no significant difference, while the Mg²⁺/Ca²⁺ ratio 4.5 treatment showed distinct difference. The survival rates and weight gain of postlarvae increased markedly when the suitable amount of K⁺ and Ca²⁺ was added to test water, and arrived at their maximum in the Na⁺/K⁺ ratio 20-30 or Mg²⁺/Ca²⁺ ratio 4.5 treatment, having no significant difference compared with normal seawater. Therefore, considering the Na⁺/K⁺, Mg²⁺/Ca²⁺ ratios and the absolute concentration of Mg²⁺, Ca²⁺ in the experimental saline groundwaters applied to Marsupenaeus japonicus farming, it should be modulated to around 30, 4.5 and 1312 mg/l, 291 mg/l, respectively.     

Effects of dietary supplementation of A3α‐peptidoglycan on the growth,immune response and defence of sea cucumber Apostichopus japonicus

To determine the effects of A3α‐peptidoglycan (A3α‐PG) extracted from Bifidobacterium sp. on the growth, immune response and disease resistance of sea cucumber Apostichopus japonicus, a 70‐day feeding trial was conducted in this study. A total of 216 sea cucumbers were fed with four practical diets prepared from a commercial feed with different contents (0, 1.5, 2.5 and 4.0 g kg^?1) of A3α‐PG. The specific growth rate (SGR), total coelomocyte count (TCC), phagocytotic activity and activities of four immunological enzymes in both cell‐free coelomic fluid (extracellular, EC) and coelomocyte lysate supernatant (intracellular, IC), including acid phosphatase (ACP), alkaline phosphatase (ALP), peroxidase (POD) and superoxide dismutase (SOD), were measured at the end of the feeding trial. Finally, the animals were administered a 16‐day Vibrio splendidus challenge via intraperitoneal injection to test the potency of A3α‐PG on disease resistance. Compared with the control (0 g kg^?1 A3α‐PG), a significant increase (P < 0.05) in SGR was observed in the groups fed with 1.5 and 2.5 g kg^?1 A3a‐PG. The TCC, ranging from 7.25 × 10⁶ to 1.05 × 10⁷ cells mL^?1, was not significantly affected (P > 0.05) by A3α‐PG,. Coelomocyte phagocytotic activities in all of the A3α‐PG‐supplemented groups were significantly activated (P < 0.05), but no significant difference (P > 0.05) was observed. Sea cucumbers fed with 1.5 and 2.5 g kg^?1 A3α‐PG exhibited significant activation (P < 0.05) of EC/IC‐ACP, EC/IC‐ALP, and EC/IC‐POD activities. A significant increase in EC‐SOD activities (P < 0.05) was exhibited by all groups with A3α‐PG supplementation. The challenge test showed that animals fed with diets containing 2.5 and 4.0 g kg^?1 A3α‐PG had significantly lower cumulative mortalities compared with the control 16 days after exposure. All of the results presented here show that A3α‐PG can positively enhance the growth, immune response and disease resistance of sea cucumber, suggesting that dietary supplementation of A3α‐PG has potential applications in the health management of economic species of sea cucumber.     

A3α‐peptidoglycan extracted from Bifidobacterium sp. could enhance immunological ability of Apostichopus japonicus

To assess the effects of A3α‐peptidoglycan (A3α‐PG) extracted from Bifidobacterium sp. on the immune response and disease resistance of sea cucumber, different concentrations (0, 0.5, 5 and 50 mg mL^?1) of A3α‐PG suspensions were used to perform hypodermic injection on Apostichopus japonicus, followed by a Vibrio splendidus challenge. Total coelomocyte count (TCC), phagocytosis activity and activities of four immunological enzymes in both cell‐free coelomic fluid (extra‐cellular, EC) and coelomocyte lysate supernatant (intracellular, IC), including acid phosphatase (ACP), alkaline phosphatase (ALP), superoxide dismutase (SOD) and peroxidase (POD), were measured at 2, 6, 14 and 24 h post injection (hpi). The TCC was not significantly affected (P > 0.05) by A3α‐PG, ranging from 1.84 × 10⁶ to 3.53 × 10⁶ cells mL^?1. The coelomocyte phagocytosis activity was significantly activated (P < 0.05) in all the A3α‐PG treatments, whereas no significant difference was observed between them except 24 hpi (P > 0.05). The EC‐ACP activity in the 5.0 mg mL^?1 treatment increased significantly (P < 0.05) at all sampling times, while the IC‐ACP activity in the 50 mg mL^?1 treatment increased significantly (P < 0.05) at 2 hpi. Also, the 5.0 mg mL^?1 treatment had significant (P < 0.05) increase in the EC‐ALP activity within 14 hpi and the EC‐POD activity at 2 hpi, respectively, while significantly (P < 0.05) enhanced IC‐ALP and IC‐POD activities were observed in the 50 mg mL^?1 treatment within 6 hpi and at 2 hpi, respectively. Only the 5.0 mg mL^?1 treatment showed significant (P < 0.05) increase in the EC‐SOD activity at 2 hpi and IC‐SOD activity within 14 hpi, respectively. The challenge test showed that the animals treated with 50 mg mL^?1 of A3α‐PG had notably lower cumulative mortality after 14 days following V. splendidus exposure. All together, these results suggest that A3α‐PG could positively enhance immune response that effectively promotes the health status of A. japonicus against V. splendidus infection.     

Purification and characterization of alkaline phosphatase from the gut of sea cucumber Stichopus japonicus

An alkaline phosphatase was purified from the gut of sea cucumber Stichopus japonicus by n-butyl alcohol extract, ammonium sulfate precipitation, ion exchange chromatography with diethylaminoethyl cellulose, gel filtration chromatography with Sephacryl S-200 and preparative electrophoresis with polyacrylamide gel electrophoresis. The native enzyme was estimated to be 166 ± 9 kDa and produced a single predominant band corresponding to active enzyme on nondenaturing electrophoresis, but showed 2 bands of 97 and 35 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting that the native enzyme is composed of two dissimilar subunits. The enzyme displayed maximum activity at pH 11 and 40 °C, showing narrow pH stability (pH 10–12) and thermal instability at temperature higher than 30 °C. The activity of the purified alkaline phosphatase was enhanced by Mg²⁺, whereas inhibited by Zn²⁺, Ca²⁺ and EDTA at 1 and 10 mM, suggesting its activity is in a magnesium ion-dependent manner. The product-analog WO₄ ^2? and product HPO₄ ^2? showed strong inhibitory effects on the enzyme activity. Using p-nitrophenyl phosphate as substrate, the V _max and K _m values were 24.45 μmol/L min and 5.76 mM, respectively.     

亚硒酸钠对刺参免疫反应中体腔液酶活力的影响

为了合理评价硒对水产养殖刺参的作用,实验检测和观察了室内模拟亚硒酸钠处理及病菌感染条件下刺参体腔液免疫酶的变化、组织病理变化和相对保护率。结果表明,随着亚硒酸钠处理强度的增加,刺参体腔液中超氧物岐化酶(SOD)、谷胱甘肽过氧化物酶(GP-X)、溶菌酶(LYZ)、酚氧化酶(PO)及碱性磷酸酶(ALP)活性升高;在同时感染病菌的情况下,SOD、GP-X及LYZ活性表现为先升高后下降的趋势,PO和ALP活性则持续上升,但随亚硒酸钠浓度升高,ALP活性增加的幅度减少,而较高亚硒酸钠浓度时累计发病率上升,相对保护率下降。研究结果表明,适宜浓度的亚硒酸钠可增强刺参对病菌感染的免疫力,而病菌感染可以加剧亚硒酸钠胁迫对刺参造成的免疫功能损伤,所以适度利用硒可减轻刺参养殖病害的发生。     

Characterization of Functional furVs as a Biomarker for Detection of Aquatic Pathogen Vibrio splendidus

Vibrio splendidus is an important opportunistic pathogen that can infect a broad range of aquatic animals including Apostichopus japonicus, leading to skin ulceration syndrome. In this study, fur gene of V. splendidus (fur_Vs), the gene encoding ferric uptake regulator (Fur) of V. splendidus, was cloned and characterized. fur_Vs possessed the signature ammonia acids of Vibrio sp., and its mRNA level was regulated by iron and coelomic fluid of A. japonicus. While the amino acid sequence of the fur_Vs showed high homology with the compared Fur proteins of other Vibrio sp., the nucleotide sequence of fur_Vs differed greatly from that of the other fur genes from Vibrio sp., based on which a pair of specific primers VSFurRTF3 and VSFurRTR3, was designed. A distinct DNA band with a length of 223 bp can be amplified from V. splendidus with great specificity. The detection limits for V. splendidus in seawater, homogenized tissues of epidermis, muscles, and coelomic fluid from A. japonicus, was 1 × 10², 1 × 10⁵, 1 × 10³, and 1 × 10² CFU/mL, respectively. Moreover, the detection limit was as low as 8 × 10^?3 pg/μL when using DNA of V. splendidus as a template. Our results provide a simple and rapid polymerase chain reaction method for the sensitive and specific detection of V. splendidus.     

饲料中添加精氨酸对仿刺参幼参生长、免疫能力及消化酶活力的影响

为研究精氨酸对仿刺参幼参生长、免疫能力及消化酶活力的影响,在基础饲料中添加0、1.00%、2.00%、3.00%和4.00%的包膜精氨酸(精氨酸含量为40%),配制精氨酸含量为0.32%、0.73%、1.16%、1.61%和1.99%的5种实验饲料(命名为D1,D2,D3,D4和D5),饲喂初始体质量9.10 g的仿刺参幼参60 d。结果表明,精氨酸显著提高了仿刺参的增重率、特定生长率和蛋白质效率,且在D4组达到最高;D3、D4和D5组仿刺参的脏壁比、肠壁比及肠长比显著低于D1和D2组,但3组之间差异不显著;精氨酸显著提高了体壁粗蛋白的沉积,但对水分及粗脂肪含量影响不显著;精氨酸显著提高了体壁谷氨酸、精氨酸、亮氨酸、组氨酸、羟脯氨酸、必需氨基酸及总氨基酸的含量,降低了赖氨酸含量;体腔液中总超氧化物歧化酶(T-SOD)、总抗氧化能力(T-AOC)、一氧化氮(NO)、一氧化氮合成酶(NOS)、碱性磷酸酶(AKP)、谷丙转氨酶(ALT)及谷草转氨酶(AST)活力随着精氨酸含量的增加而显著提高;肠道蛋白酶活力与饲料精氨酸含量呈二次负相关关系(Y酶活=-3.228X2精氨酸+2.865X精氨酸+26.24,R2=0.934),D3、D4和D5组纤维素酶活力显著高于D1和D2组,淀粉酶及脂肪酶活力不受精氨酸含量的影响。以增重率为评价指标,一元二次回归分析表明,体质量9.10 g的仿刺参饲料中精氨酸适宜含量为1.55%(7.10%饲料粗蛋白)。     

Vitamin E requirement of sea cucumber (Apostichopus japonicus) and its' effects on nonspecific immune responses

This feeding trial was conducted to determine the vitamin E requirement of sea cucumber (Apostichopus japonicus) and to study the effects of vitamin E on nonspecific immune parameters. Commercial sea cucumber diets supplemented with 0, 50, 100, 200, 500 or 1000 mg kg^?1 vitamin E were fed to juvenile sea cucumber (7.96 ± 0.01 g) for 60 days. Specific growth rate and total coelomocyte counts were maximized in sea cucumber fed diets supplemented with 100 mg kg^?1 vitamin E. Reactive oxygen species production was maximized at 50 mg kg^?1 vitamin E, whereas superoxide dismutase activity was the highest in sea cucumber fed diets supplemented with 100 mg kg^?1 vitamin E. The vitamin E concentration in the body wall of the sea cucumber increased as the vitamin E level in the diet increased. Analysis of specific growth rate and total coelomocyte counts of the sea cucumber indicated that 88–92 mg kg^?1 vitamin E is required for maximal growth and nonspecific immune responses of A. japonicus and that 114.7 mg kg^?1 vitamin E is required to minimize lipid peroxidation levels.     

Supplementation of potassium, magnesium and sodium chloride in practical diets for the Pacific white shrimp, Litopenaeus vannamei, reared in low salinity waters

The culture of Litopenaeus vannamei in inland low salinity waters is currently being practiced in various countries around the world. These environments are often deficient in key ions essential for normal physiological function, including potassium (K⁺) and magnesium (Mg²⁺). Farmers have sometimes been able to counteract ionic deficiencies in the water profile by adding mineral salts containing sources of K⁺ and Mg²⁺. The purpose of this study was to explore the possibility of correcting deficiencies of K⁺ and Mg²⁺ in the water profile with dietary supplementation of these minerals. Two separate 7‐week experiments were conducted in 4.0 g⁻¹ artificial low salinity water to evaluate the effects of mineral supplements (K⁺, Mg²⁺ and NaCl) to diets of L. vannamei reared in low salinity waters. In trial 1 seven diets were formulated (10 g NaCl kg⁻¹, 20 g NaCl kg⁻¹, 150 mg kg⁻¹ Mg²⁺, 300 mg kg⁻¹ Mg²⁺, 5 g K⁺ kg⁻¹, 10 g K⁺ kg⁻¹, and a basal diet to serve as a control). Minerals were added in the form of purified potassium chloride (KCl), magnesium chloride (MgCl₂·6H₂O) and NaCl. Trial 2 evaluated the use of a coating agent for the Mg²⁺ and NaCl treatments, while a K⁺ amino acid complex was utilized in the K⁺ treatments to reduce mineral leaching. Trial 2 was performed using similar treatment levels as trial 1. Shrimp survival and growth were assessed in both experiments. Results from trial 1 indicated no significant differences in survival, growth or percent weight gain. Results from trial 2 revealed no significant differences in survival and growth in the NaCl and Mg²⁺ treatments. However, significant differences in growth (P < 0.05) were observed when using the 10 g K⁺ kg⁻¹ treatment, suggesting that dietary supplementation of a K⁺ amino acid complex may help improve growth of the species in low salinity waters.