热胁迫对白梭吻鲈鳃组织结构及抗氧化指标的影响

温度是导致鱼类生理变化的最重要的环境因素之一。热应激导致氧化应激,破坏抗氧化防御系统,并导致脂质和组织病理学的损害。白梭吻鲈(Sander lucioperca)属于亚冷水性鱼类,最适生长水温20～24℃。近年来,由于夏季气温极高,中国池塘出现了大量的白梭吻鲈死亡,白梭吻鲈的水产养殖业一直面临着高温的严峻挑战。鱼鳃参与鱼类许多重要功能,如呼吸,渗透调节和排泄,与外部环境保持密切接触,对水质变化特别敏感。为探究热胁迫对白梭吻鲈鳃组织结构及抗氧化指标的影响,将100尾鱼暴露于28℃,31℃,34℃,36℃热应激2h,以23℃作为对照组。收集鳃用于石蜡切片制备,并将苏木精伊红（HE）染色用于一般组织学观察。测定超氧化歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GPX）活性和丙二醛（MDA）等抗氧化指标。结果显示,28 ℃组鳃小片发生充血,31 ℃组部分鳃小片末端卷曲变形、上皮细胞水肿,34 ℃组鳃小片变形加重,且有少量融合,上皮细胞水肿加重、明显隆起,鳃小片之间粘液细胞肥大、增生,36 ℃组相邻鳃小片大量融合且严重变形,卷曲混乱排列,大量上皮细胞坏死、脱落,鳃丝粘液细胞增生、肥大;随着胁迫温度升高,鳃中SOD、CAT、GPX、MDA均整体呈现先升高、后降低的变化趋势,且在36℃时显著下降。这表明在热应激后,鱼体内ROS增加,脂质过氧化增加。由于体内产生大量的ROS和H2O2,机体也增加了GPX和CAT的合成。当暴露在36℃时,胁迫强度超过白梭吻鲈的适应能力,抗氧化系统功能无法正常进行。表明热胁迫对白梭吻鲈鳃组织有损伤,且随着热胁迫温度的升高,其鳃受损情况越发严重。     

饲料溶菌酶添加水平对氨氮应激下吉富罗非鱼血清生化指标、抗菌性能和肝脏抗氧化能力的影响

为研究氨氮应激下溶菌酶对吉富罗非鱼血清生化指标、抗菌性能及肝脏抗氧化能力的影响,用6种不同溶菌酶添加水平(0、18、36、54、72和90 mg/kg)的饲料(分别记为L0、L18、L36、L54、L72和L90)饲喂初始均重为(11.35±0.08)g的罗非鱼60 d,之后应用氯化铵进行24 h氨氮应激实验。结果显示:1应激后,各组鱼的血清生化指标在组间存在较大差异(P0.05),不同溶菌酶添加水平下的鱼体对氨氮应激产生了不同的响应机制。L54组鱼主要通过激发免疫系统并调节蛋白质代谢来抵抗氨氮浓度突变;L72和L90组鱼主要通过调节高密度脂蛋白和低密度脂蛋白、胆固醇和甘油三酯之间的动态变化来缓解氨氮对机体的应激。2血清抗菌实验表明,L54和L72组鱼对大肠杆菌、金黄色葡萄球菌和嗜水气单胞菌的抑制能力最大,且显著高于对照组(P0.05);L36~L90组对溶藻弧菌的抑制作用显著高于对照组和L18组(P0.05);各溶菌酶添加组的鱼对枯草芽孢杆菌具有不同程度的保护作用。3应激后肝脏的超氧化物歧化酶、谷胱甘肽过氧化物酶和过氧化氢酶活性随溶菌酶添加水平的增加总体呈现先升后降的变化趋势,L54组显著高于对照组(P0.05);各添加组丙二醛含量均显著低于对照组(P0.05)。研究表明,54和72mg/kg溶菌酶添加水平对氨氮应激下吉富罗非鱼的血清生化指标、抗菌活性和肝脏抗氧化指标产生了最积极有效的调控,能够增强鱼体的抗应激能力。     

盐胁迫对雨生红球藻虾青素累积、虾青素合成相关酶基因表达和抗氧化指标的影响

为探讨盐胁迫对雨生红球藻(Haematococcus pluvialis)虾青素合成的影响与机理,以及雨生红球藻各抗氧化机制之间的关系,本研究采用生化和分子生物学方法研究了不同浓度(0.04 mol/L、0.08 mol/L、0.12 mol/L和0.16 mol/L)和不同时间(3 d、6 d和9 d)的盐(Na Cl)胁迫对雨生红球藻生长、虾青素积累、番茄红素β-环化酶(Lcy)、β-胡萝卜素羟化酶(Crt R-B)和β-胡萝卜素酮化酶(Bkt)基因表达、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性以及丙二醛(MDA)含量的影响。结果表明,各胁迫时间的雨生红球藻的密度均随着盐胁迫浓度的增加而不断下降,在盐胁迫的第9天,雨生红球藻的死亡率和孢子比例均随着盐胁迫浓度的增加而不断升高;雨生红球藻虾青素含量、Lcy、Crt R-B和Bkt基因表达量均随着盐胁迫浓度和时间的增加而不断提高;雨生红球藻SOD、CAT和GSH-Px活性以及MDA含量在不同浓度和不同时间的盐胁迫下与对照组(0.00 mol/L Na Cl)相比均升高,且在不同时间的0.12 mol/L Na Cl胁迫下与对照组相比均显著升高(P0.05);雨生红球藻虾青素含量、Lcy、Crt R-B和Bkt基因表达量在盐胁迫的早期(第3天)和中期(第6天)阶段较低,在盐胁迫的后期(第9天)阶段较高,而SOD、CAT和GSH-Px活性以及MDA含量在盐胁迫的早期和中期阶段较高,在盐胁迫的后期阶段较低。实验结果说明了适当浓度和时间的盐胁迫能促进雨生红球藻累积虾青素,雨生红球藻在盐胁迫下主要是通过提高虾青素合成相关酶基因的转录水平来促进虾青素的合成,其虾青素和抗氧化酶的抗氧化活性可能互为补充,共同保护雨生红球藻免受盐胁迫的氧化损伤。     

Effects of dietary protein to energy ratios on growth,body composition and digestive enzyme activities in Chinese mitten‐handed crab,Eriocheir sinensis

This study aimed to determine the optimal protein to energy ratio (P/E ratio) and evaluate the effect of dietary protein and lipid levels on growth performance, body composition and digestive enzymes activities in Chinese mitten‐handed crab, Eriocheir sinensis. Nine practical diets containing three levels both for protein (DP 30%, 35% and 40%) and lipid (DL 2%, 7% and 12%) with P/E ratios ranging from 13.69 to 19.79 mg KJ^?1 were fed to four replicates of crabs (3.39 ± 0.10 g) for 10 weeks. Weight gain increased significantly with the increase in DP level at each DL level. Moreover, weight gain increased in crabs fed with diets containing DL level from 2% to 12% and DP level from 30% to 35%. However, the diet containing 40% DP and 12% DL levels significantly decreased the growth performance and protein efficiency of the crabs. The whole crab and hepatopancreas lipid contents also increased as dietary lipid increased, but not dietary protein. The total protease activity increased significantly with the increase in dietary protein at each lipid level. The lipase activity was statistically comparable among different DL levels at each DP level. Taken together, the crab fed the diet containing 35% protein and 12% lipid levels with P/E 15.77 mg KJ^?1 revealed optimal growth, feed utilization efficiency and digestive enzymes activities. Moreover, our study indicated that the higher dietary lipid level at a relatively lower dietary protein level could provide protein sparing effect in Eriocheir sinensis.     

超低温冷冻保存对大黄鱼精子酶活性的影响

研究超低温(-196℃)冷冻保存对大黄鱼(Pseudosiaena crocea)精子内总ATP酶、肌酸激酶(CK)、琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)等酶活性的影响。运用试剂盒分别测定了冷冻前后大黄鱼精子内酶活性的变化。结果表明,经过超低温冷冻保存后,大黄鱼精子的活力下降,精子内GR活性从(4.42±0.29)U·L-1增加到(58.93±2.26)U·L-1(P<0.05);其它几种酶的活性均显著下降(P<0.05),总ATP酶、CK、SDH的活性分别从冻前的(60.16±5.88)U·mL-1、(11.91±0.76)U·mL-1和(51±2.16)U·mL-1下降到(3.54±0.37)U·mL-1、(10.22±0.32)U·mL-1和(31.5±2.08)U·mL-1;LDH、SOD和CAT活性从冻前的(7 806.44±110.11)U·L-1、(42.65±1.56)U·mL-1和(119.91±8.10)U·mL-1下降到(2 654.13±70.06)U·L-1、(31.99±1.57)U·mL-1和(55.87±2.32)U·mL-1。超低温冷冻保存对大黄鱼精子活力和精子酶活性均有显著影响。     

3种水质调控方式下参池沉积物酶活性的比较研究

2015年10月至2016年9月对自然纳潮、微孔曝气、养水机3种水质调控方式下海参池塘沉积物中淀粉酶、蛋白酶、碱性磷酸酶、脱氢酶的活性进行了跟踪比较研究。研究结果显示,3种水质调控方式下池塘沉积物中的淀粉酶活性年变化为0.126~0.880 mg/g,年均值(0.410±0.180) mg/g,蛋白酶活性年变化为0.024~0.472 mg/g,年均值(0.190±0.103) mg/g,碱性磷酸酶活性年变化为0.068~1.042 mg/g,年均值(0.340±0.196) mg/g,脱氢酶活性年变化为12.092~52.794 mL/g,年均值(26.980±8.295) mL/g。3种水质调控方式下池塘沉积物中蛋白酶、碱性磷酸酶及脱氢酶活性均在自然纳潮池塘中均值最高,变化幅度最大;淀粉酶活性均值则在自然纳潮池塘中最低,养水机池塘最高,这与养水机池塘有机质最低,自然纳潮池塘最高,养水机池塘沉积物的细菌多样性最高,真菌数量最多有关。表明养水机能够快速去除沉积物中氮、磷有机化合物,有利于池塘的正常物质循环。本研究从沉积物酶活性的角度,探讨了养水机的作用效果与其他两种水质调控方式产生差异的机理。     

皱纹盘鲍食道的结构与功能

以组织学，组织化学，扫描电镜和透射电镜观察及酶活性测定等方法研究了皱纹盘鲍的食道，食道可分为前，中后，三段，中段又可分为食物通道和食道侧囊，食道粘膜上皮由纤毛柱状细胞，粘液细胞，闰状腺细胞，微绒毛细胞和分泌细胞组成，纤毛柱状细胞参与运输食物和分泌物，并呈现吸收细胞的结构特征；粘液细胞分泌中性和酸性粘多糖；颗粒状腺细胞内充盈分泌颗粒；微绒毛细胞呈现吸收细胞的特征；分泌细胞具有强的蛋白酶等酶活性，能以楔浆分泌形式分泌消化酶，该细胞还具有吸收和细胞内消化作用。食道中段还呈现3种植物多糖酶活性。     

Effects of dietary thiamin supplement on growth,body composition and intestinal enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 1050 juvenile Jian carp (Cyprinus carpio var. Jian) (8.20 ± 0.02 g) were fed diets containing seven graded levels of thiamin (0.25, 0.48, 0.79, 1.06, 1.37, 1.63 and 2.65 mg kg^?1) for 60 days to investigate the effects of thiamin on growth, body composition and digestive enzyme activities. Percent weight gain (PWG), feed intake and feed efficiency (FE) were the lowest in fish fed the basal diet (P < 0.05). Protein productive value and lipid productive value increased with increasing dietary thiamin levels up to 1.06 and 0.79 mg kg^?1 diet, respectively (P < 0.05). Body protein and lipid increased with increasing dietary thiamin levels (P < 0.05), while moisture and ash of fish carcasses decreased with the increase in dietary thiamin supplementation (P < 0.05). Intestinal folds height had a similar trend to PWG (P < 0.05). Activities of α‐amylase, lipase, trypsin, Na⁺, K⁺‐ATPase, alkaline phosphatase and gamma‐glutamyl transpeptidase in intestine were all affected by the dietary thiamin (P < 0.05). In conclusion, thiamin could improve growth and intestinal enzyme activities of juvenile Jian carp. The dietary thiamin requirement of juvenile Jian carp (8.0–60.2 g) based on PWG was 1.02 mg kg^?1 diet.     

Effects of genetically modified and non‐genetically modified soybeans with different heat treatments on growth and health of Cyprinidae species with different feeding habits

An 8‐week growth trial was conducted to investigate the effects of non‐genetically modified (nGM) soybean (Youchun 06‐1) and genetically modified (GM) soybean (Roundup Ready^®) with and without a heat treatment on the growth and health of three Cyprinidae species with different feeding habits (grass carp Ctenopharyngodon idellus, gibel carp Carassius auratus gibelio, and black carp Mylopharyngodon piceus; body weight: 283.0 ± 2.0 g, 60.5 ± 0.7 g, and 261.4 ± 3.1 g). Five diets (FM, nGMS, hnGMS, GMS, and hGMS) were made with fishmeal and full‐fat soybean (Youchun 06‐1, heat‐treated Youchun 06‐1, Roundup Ready^®, and heat‐treated Roundup Ready^®). The FM diet contained fishmeal as the sole protein source. The experimental diets contained full‐fat soybean meal as 60% of dietary protein. Both temperature (60, 80, 100, and 120°C) and duration (1 and 2 hr) of heating influenced trypsin inhibitor activity (TIA) and protein solubility of nGM soybean and GM soybean. The TIA in GM soybean was higher than that in nGM soybean. After heating at 120°C for 2 hr, each amino acid of soybean treatment showed a slight decline. Neither source of soybean nor heat treatment affected the growth performance, feed utilization, chemical composition of the whole fish body and muscle, and lipase or amylase activity of the three Cyprinidae species (p > 0.05). Heat treatment of soybean meal slightly increased the plasma antioxidant capacity of the three fish and plasma cholecystokinin of black carp and grass carp. The unheated soybean treatment adversely affected the height and density of black carp intestinal villi, and all the soybean treatments caused disruption of the grass carp intestinal epithelium.     

我国尼罗罗非鱼和奥利亚罗非鱼养殖群体的遗传渐渗

用ＬＫＢ平板电泳仪，４．４％聚丙烯酰胺凝胶对南京罗非鱼良种场尼罗罗非鱼，奥利亚罗非鱼养殖群体肌，肝，脑，心，眼中的１０种同工酶进行电泳分析的结果表明，奥利亚罗非鱼的群体未见的多态位点，平均杂合度为０，是超“纯”的养殖群体，尼罗罗非鱼群体中有２０％的尼奥杂交鱼，其体形酷似尼罗，难以肉眼鉴别，由此可见，我国尼罗罗非鱼养殖群体中已存在遗传渐渗问题，须注意防杂和提纯。在剔除了杂交鱼后，该场尼罗罗非鱼群体的