Influence of glutamine and vitamin E on growth and antioxidant capacity of fish enterocytes

The present study explored the effect of glutamine and vitamin E on growth and anti-oxidation capacity of isolated fish enterocytes. Fish enterocytes were cultured with six medium, respectively, containing 0, 2.0, 4.7, 6.8, 8.1, 9.2 mmol L⁻¹ glutamine for 64 h. The results showed that glutamine could promote fish enterocytes proliferation and differentiation. Fish enterocytes were cultured with different medium containing 0, 2.5, 3.5, 4.5, 6.0, 7.0 μg mL⁻¹ vitamin E for 96 h. The results showed that cells proliferation and differentiation were not significantly enhanced, but anti-superoxide anion activity, anti-hydroxy radical activity, reduced glutathione concentration, the ratio between reduced and total glutathione in the cells were significantly enhanced, and the malondialdehyde concentration in the culture medium was significantly depressed with the vitamin E treatment. In the whole, the present results firstly indicated that glutamine could promote fish enterocytes growth, but vitamin E could not. Vitamin E could promote fish enterocytes antioxidant capacity and cellular structural integrity. These data would be instructive for glutamine and vitamin E supplement in aquaculture diets.     

Preventive and reparative effects of isoleucine against copper-induced oxidative damage in primary fish enterocytes

The present study aimed to assess the possible preventive and reparative effects of isoleucine (Ile) against copper (Cu)-induced oxidative stress in fish enterocytes in vitro. In experiment 1, enterocytes were preincubated with increasing concentrations of Ile (0, 50, 120, 190, 260, and 330 mg L^?1) for 72 h followed by exposure to 6 mg L^?1 Cu for 24 h. In experiment 2, the enterocytes were pretreated with 6 mg L^?1 Cu for 24 h and then treated with 0–330 mg L^?1 Ile for 72 h to investigate its potential reparative role. The results of experiment 1 showed that Cu exposure increased lactate dehydrogenase (LDH) activity and malondialdehyde and protein carbonyl (PC) content; these changes were completely suppressed by pretreatment with Ile at optimum concentrations (P < 0.05). Moreover, Ile pretreatment prevented the decrease in superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities in the enterocytes exposed to Cu (P < 0.05). Additionally, cells exposed to Cu exhibited adaptive increases in glutathione-S-transferase (GST) activity. In experiment 2, the LDH activity and protein oxidation induced by Cu were completely reversed by Ile posttreatment. Meanwhile, the Cu-induced decrease in SOD, GPx, and GST activity was completely reversed by subsequent Ile treatment, but the reduced glutathione content was not restored. Collectively, these results indicate that Ile suppresses Cu-induced oxidative damage via preventive and reparative pathways in primary enterocytes and thus protects the structural integrity of enterocytes in fish.     

Luminal uptake of Vibrio (Listonella) anguillarum by shed enterocytes – a novel early defence strategy in larval fish

As adhesion and translocation through fish gut enterocytes of the pathogen Vibrio (Listonella) anguillarum are not well investigated, the effective cause of disease and mortality outbreaks in larval sea bass, Dicentrarchus labrax, suffering from vibriosis is unknown. We detected V. anguillarum within the gut of experimentally infected gnotobiotic sea bass larvae using transmission electron microscopy and immunogold labelling. Intact bacteria were observed in close contact with the apical brush border in the gut lumen. Enterocytes contained lysosomes positive for protein A‐gold particles suggesting intracellular elimination of bacterial fragments. Shed intestinal cells were regularly visualized in the gut lumen in late stages of exposure. Some of the luminal cells showed invagination and putative engulfment of bacterial structures by pseudopod‐like formations. The engulfed structures were positive for protein A‐colloidal gold indicating that these structures were V. anguillarum. Immunogold positive thread‐like structures secreted by V. anguillarum suggested the presence of outer membrane vesicles (MVs) hypothesizing that MVs are potent transporters of active virulence factors to sea bass gut cells suggestive for a substantial role in biofilm formation and pathogenesis. We put forward the hypothesis that MVs are important in the pathogenesis of V. anguillarum in sea bass larvae.     

纳米氧化锌对斑马鱼肠组织的氧化损伤

为探讨纳米氧化锌颗粒(ZnO-NPs)对斑马鱼肠组织的损伤及损伤机理,研究了ZnO-NPs对肠组织结构、抗氧化酶活性及相关凋亡基因表达的影响。将斑马鱼分别暴露于浓度为0、0.05、0.1、5、10、25和50 mg/L的ZnO-NPs水体中,在4、24和96 h时,用比色法测定了肠组织中丙二醛(MDA)含量、谷胱甘肽过氧化物酶(GSH-PX)、谷胱甘肽硫转移酶(GST)活性;用实时荧光定量法测定了Bcl-2、Bax、p53和MDM2 mRNA表达水平的变化;在第7、15和30天时用H.E染色技术观察了肠组织解剖结构变化。与对照组相比,实验组结果显示:斑马鱼肠中的MDA含量均高于对照组;随着时间的延长,组织中GSH-PX和GST活性呈先升高后下降的趋势;肠组织中Bax/Bcl-2和p53的mRNA的表达量均升高,而MDM2的mRNA表达量则随着时间的延长和浓度的增加表现为先降低后升高;H.E染色观察到肠组织结构发生变化,表现为杯状细胞增多,肿胀变形,部分细胞质空泡化,肠中淋巴细胞增多,肠绒毛侵蚀变形,且有时间和浓度依赖性。结果表明:ZnO-NPs能诱导斑马鱼肠组织产生氧化应激作用,使组织中抗氧化酶活性发生变化,诱导肠中细胞凋亡相关基因的表达,并且能对肠组织结构造成损伤。     

Microcystin-RR induces apoptosis in fish lymphocytes by generating reactive oxygen species and causing mitochondrial damage

Microcystin-RR (MCRR), a very widespread toxic microcystin in China, has previously been proved to induce sensitive apoptosis in Carassius auratus lymphocytes in vitro. This study focused on the role of intracellular Ca²⁺, mitochondrial membrane potential, reactive oxygen species, and intracellular ATP in the mechanism of MCRR-induced apoptotic toxicity to fish lymphocytes. Compared with controls, administration of MCRR (10 nmol L⁻¹) caused a massive calcium influx resulting in elevation of reactive oxygen species (ROS), rapid disruption of mitochondrial membrane potential (ΔΨm), and depletion of ATP. This study provided a possible mechanism for the cytotoxicity of microcystins to fish lymphocytes.     

MS222 does not induce primary DNA damage in fish

The genotoxicity of the anaesthetic MS222 (tricaine) was analysed in fish under both in vivo and in vitro conditions. Based on results of the single cell gel/Comet assay, MS222 had no direct genotoxic effect on the experimental fish, indicating that MS222 does not induce primary DNA damage. These results suggest that the use of this important anaesthetic in aquaculture can be considered to be safe in terms of genotoxicity. R. E. Barreto and A. M. M. C. Gontijo contributed equally to the study.     

菲和苯并（b）荧蒽曝露对翡翠贻贝外套膜的氧化胁迫及损伤

实验室条件下研究了不同质量浓度（2.0μg.L-1、10.0μg.L-1和50.0μg.L-1）的菲（PHE）和苯并（b）荧蒽（BbF）胁迫15 d和清洁海水恢复7 d中翡翠贻贝（Perna viridis）外套膜组织中超氧化物歧化酶（SOD）活力和丙二醛（MDA）质量摩尔浓度的变化。结果表明,PHE胁迫下翡翠贻贝外套膜SOD活力呈先升高后降低的变化规律;BbF胁迫下的翡翠贻贝外套膜SOD活力在胁迫第15天时被显著诱导（P〈0.05）。从b（MDA）的变化来看,PHE和BbF均可导致翡翠贻贝外套膜明显的氧化损伤,之后在清洁海水的净化过程中,这种损伤逐渐降低并恢复至正常水平。鉴于2种PAHs胁迫下翡翠贻贝外套膜SOD活力和b（MDA）均发生明显变化并表现出一定的差异性,翡翠贻贝体内的生化指标适合指示PAHs对海洋环境的污染。     

Blood parameters and electrocardiogram in squeezed fish simulating the effect of net damage and recovery

ABSTRACT:   In the present study, rubber bands were tied around the girth of rainbow trout Oncorhynchus mykiss to simulate and assess the physiological damage and survival rates of fish that encounter gill nets. Physiological condition was assessed by analysis of blood lactate, pH, oxygen partial pressure and plasma potassium concentration before and after release from binding. Twenty-four hours after release, half of the test fish had died. Although the binding forces used to hamper fish did not differ significantly, blood lactate levels in the dead fish rose to 10-fold higher than those in the survivors. Consequently, the pH level fell in the group that died, whereas in the surviving group it fell only slightly and soon recovered. Potassium concentration increased after release from binding in fish that died, and the P o ₂ levels in these fish continued to drop until just before death. It was concluded that when fish get entangled in fishing nets, blood lactate accumulates because of physical fatigue and the elevated lactate levels lead to metabolic acidosis. Finally, fatal metabolic conditions could cause higher mortality rates, even in escapees.     

Herpesvirus cyprini: a search for viral genome in infected fish by infected fish by in situ hybridization

Abstract. Herpesvirus cyprini (CHV) genome was traced in carp, Cyprinus carpio L., after acute infection by the method of in situ hybridization with biotinylated probes. The viral genome was detected in several tissues including cranial nerve ganglia. Subcutaneous tissue and spinal nerves. However, at this stage, viral antigens were not detected and the virus was not isolated. The viral genome was also detected in the same fish tissues when papillomas were present which contained viral antigens and even infective virus particles. After papilloma regression, the viral genome was still detected in these tissues. It is suggested that CHV becomes latently established in cranial nerve ganglia, subcutaneous tissue and spinal nerves, and is associated with the induction and recurrence of papillomas.     

慢性氨氮暴露诱发黄颡鱼幼鱼谷氨酰胺积累、氧化损伤及免疫抑制的研究

为了研究慢性氨氮胁迫对黄颡鱼幼鱼生长性能、大脑谷氨酰胺积累、肝脏抗氧化酶活性、非特异性免疫应答及抗嗜水气单胞菌感染能力的影响,实验挑选初始体质量为(1.94±0.05)g的健康黄颡鱼幼鱼180尾,开展为期56 d的慢性氨氮胁迫实验.结果表明,实验鱼的终末体质量、增重及饲料效率,氨氮组与对照组无显著性差异,但肝体比氨氮组显著高于对照组;氨氮组实验鱼大脑中氨氮和谷氨酰胺含量显著高于对照组,但谷氨酸含量各组间无显著性差异;氨氮组实验鱼肝脏中超氧化物歧化酶、过氧化物酶及谷胱甘肽过氧化物酶活性显著低于对照组,但硫代巴比妥酸反应物含量氨氮组显著高于对照组;氨氮组实验鱼肝脏中溶菌酶活性、头肾巨噬细胞吞噬指数和呼吸爆发显著低于对照组;感染嗜水气单胞菌14d后,氨氮组和对照组实验鱼累计死亡率无显著性差异.研究表明,黄颡鱼幼鱼遭受亚致死浓度的慢性氨氮胁迫,能够导致大脑中谷氨酰胺含量升高;氨氮组黄颡鱼肝脏中硫代巴比妥酸反应物的过度积累表明,应激产生的大量自由基并不能被机体自身的抗氧化酶体系完全清除;亚致死浓度的慢性氨氮胁迫会对黄颡鱼幼鱼的免疫应答体系造成抑制.     

The effects of dietary thiamin on oxidative damage and antioxidant defence of juvenile fish

The present study explored the effects of thiamin on antioxidant capacity of juvenile Jian carp (Cyprinus carpio var. Jian). In a 60-day feeding trial, a total of 1,050 juvenile Jian carp (8.20 ± 0.02 g) were fed graded levels of thiamin at 0.25, 0.48, 0.79, 1.06, 1.37, 1.63 and 2.65 mg thiamin kg^?1 diets. The results showed that malondialdehyde and protein carbonyl contents in serum, hepatopancreas, intestine and muscle were significantly decreased with increasing dietary thiamin levels (P < 0.05). Conversely, the anti-superoxide anion capacity and anti-hydroxyl radical capacity in serum, hepatopancreas, intestine and muscle were the lowest in fish fed the thiamin-unsupplemented diet. Meanwhile, the activities of catalase (CAT), glutathione peroxidase, glutathione S-transferase and glutathione reductase, and the contents of glutathione in serum, hepatopancreas, intestine and muscle were enhanced with increasing dietary thiamin levels (P < 0.05). Superoxide dismutase (SOD) activity in serum, hepatopancreas and intestine followed a similar trend as CAT (P < 0.05). However, SOD activity in muscle was not affected by dietary thiamin level (P > 0.05). The results indicated that thiamin could improve antioxidant defence and inhibit lipid peroxidation and protein oxidation of juvenile Jian carp.     

龙须菜在网箱养殖区的生物修复研究

在福建省东山县八尺门网箱养殖区的浅水区吊养面积约0.7 hm2的龙须菜(Gracilaria lemaneiformis),研究龙须菜对网箱养殖区水质的修复效果。通过对实验区及其内外两侧的定点监测,以及对与潮流方向一致和垂直的2个断面的监测,结果表明,修复区的溶解氧(DO)浓度明显高于非修复区,无机氮(IN)、无机磷(IP)浓度低于非修复区,由此可见,通过海水交换,龙须菜可以吸收流经修复区的IN、IP,并提高流出修复区的海水的DO水平。这有利于改善网箱养殖区的海水水质,提高网箱养鱼的成活率和生长率,促进海水养殖的可持续发展。[中国水产科学,2007,14(3):488-492]     

Pinger affects fish catch efficiency and damage to bottom gill nets related to bottlenose dolphins

There is some evidence that the presence of Tursiops truncatus in fishing areas represents a real economic threat to fishermen due the dolphin feeding on the entangled fish, damaging the nets and reducing the fish catch. We have carried out experiments to assess the efficiency of a pinger in decreasing the interaction between the dolphins and fishing nets, in a fishing area off the coast of southern Italy, where Tursiops truncatus is frequently observed to interact with bottom gill nets. Two identical monofilament bottom gill nets (900 m long), one equipped with pingers and the other without, were used to measure the effect of these pingers on the abundance of the catch and net damage. For each haul (58 in total), data on dolphin sightings near the nets, damage judged to have been done by dolphins, weight and species composition of the catch were collected. All damage to the nets were recorded at the end of each haul. Dolphins in the fishing area were sighted 11 times out of 29 fishing activities (38%). The net equipped with pingers contained 28% more fish (biomass) than the net without pingers (t test, P < 0.04) and was less damaged (−31%, t test, P < 0.01). To assess whether the efficacy of these pingers remain constant over long period, long-term experiments should be carried out.     

高糖饲料对吉富罗非鱼生长性能、饲料利用和糖脂代谢的影响

为了从生长性状、器官健康和营养代谢水平方面探究草食性鱼类草鱼 (Ctenopharyngodon idella) 对高糖饲料的适应和利用能力,以初始体质量 (132.01±16.43) g的草鱼为研究对象,分别配制正常糖水平 (15%,对照组) 和高糖水平 (40%,高糖组) 的等氮等能饲料分组投喂草鱼140 d,每组设3个重复,每个重复300 尾鱼。结果显示,相较于对照组,第60、第80 和第140天时高糖组草鱼的体质量显著增加 (P<0.05),第140天时,高糖组草鱼的脏体比、肠系膜脂肪系数、中肠绒毛高度和肝糖原含量均显著提高且肝脏和中肠肠道组织出现损伤。随实验时间的延长,高糖组葡萄糖激酶 (gk)、丙酮酸激酶 (pk)、糖原合酶 (gys2)、脂肪酸合成酶 (fas) 基因表达量呈下降趋势,磷酸烯醇式丙酮酸激酶 (pepck)、葡萄糖-6-磷酸酶 (g6pase) 基因表达量呈上升趋势,对照组gys2、fas、pepck和g6pase基因表达量变化趋势与高糖组一致,但gk、pk基因表达量在各时间点均无显著性变化。第140天时,与对照组相比,高糖组pk基因表达量显著下调,g6pase、pepck和fas基因表达量显著上调。综上,采用高糖饲料投喂草鱼可促进其体质量增加,但长期饲喂会对其生理指标造成一定负面影响,肝脏糖代谢相关基因表达变化显著。

     

A new high in fish production in India with record yields by composite fish culture in freshwater ponds

Composite fish culture of the Indian major carp, Catla catla, Labeo rohita and Cirrhinus mrigala together with the Chinese silver carp Hypophthalmichthys molitrix and grass carp Ctenopharyngodon idella and the Indonesian strain of common carp (Cyprinus carpio communis) was carried out in experimental ponds in 1971–1972 and 1973–1974. The management techniques adopted aimed at high yields of marketable fish (around 1 kg) in 1 year.In the first year, production varied from 3 889 to 5 600 kg/ha/year and this rose to an average of 8 200 kg/ha/year with a maximum of 9 389 kg/ha/year in the second year. The important changes made to the management schedule in 1973 were increased stocking density, greater use of feed and fertilizers, and provision of more weeds for grass carp.The experimental studies demonstrate the practicability of raising large crops of healthy marketable fish in India.     

不同碳水化合物水平对大口黑鲈鳃组织结构、抗氧化能力和免疫能力的影响

为探究多鳞白甲鱼幼鱼日粮中最佳碳水化合物添加类型,以葡萄糖、麦芽糖、糊精、玉米淀粉及纤维素5种碳水化合物为糖源,分别配制5种日粮,对多鳞白甲鱼幼鱼进行了为期56 d的养殖实验。结果显示,①生长及饲料利用率方面,麦芽糖组的终末体重、特定生长率、饵料系数以及蛋白质效率等指标最佳,纤维素组最差。②体成分方面,纤维素组全鱼粗脂肪水平最低,粗灰分水平显著高于麦芽糖组,麦芽糖组及纤维素组肌糖原含量最低,糊精组肝糖原含量最低。③主成分分析表明,C16:0、SFA、MUFA和PUFA比例对肝胰脏脂肪酸比例产生主要影响,其中,C16:0和SFA比例在麦芽糖组显著提高,MUFA比例在糊精组显著提高,而PUFA在纤维素组显著提高;C18:1n-9和MUFA比例对肌肉和脂肪组织总体脂肪酸组成产生主要影响,二者比例在麦芽糖组显著提高。④血清生化指标方面,麦芽糖组及纤维素组谷丙转氨酶活性、麦芽糖组谷草转氨酶活性显著较高,葡萄糖组及纤维素组碱性磷酸酶活性、球蛋白含量较高,而血糖含量反之。⑤抗氧化能力方面,麦芽糖组肝胰脏过氧化氢酶、总超氧化物歧化酶、谷胱甘肽过氧化物酶活性最高,纤维素组丙二醛含量最高。研究表明,麦芽糖是多鳞白甲鱼幼鱼适宜的碳水化合物源,玉米淀粉及糊精次之,纤维素及葡萄糖不是多鳞白甲鱼幼鱼适宜的碳水化合物源,实际生产中推荐玉米淀粉作为碳水化合物源。本研究将为多鳞白甲鱼专用饲料研发提供重要参考。     

Learning of foraging skills by fish

This chapter outlines the relationships between a number of key factors that influence learning and memory, and illustrates them by reference to studies on the foraging behaviour of fish. Learning can lead to significant improvements in foraging performance in only a few exposures, and at least some fish species are capable of adjusting their foraging strategy as patterns of patch profitability change. There is also evidence that the memory window for prey varies between fish species, and that this may be a function of environmental predictability. Convergence between behavioural ecology and comparative psychology offers promise in terms of developing more mechanistically realistic foraging models and explaining apparently ‘suboptimal’ patterns of behaviour. Foraging decisions involve the interplay between several distinct systems of learning and memory, including those that relate to habitat, food patches, prey types, conspecifics and predators. Fish biologists, therefore, face an interesting challenge in developing integrated accounts of fish foraging that explain how cognitive sophistication can help individual animals to deal with the complexity of the ecological context.     

投饵网箱养鱼促进温杖子水库渔业高速增产的生态经济效益分析

温杖子水库按养鱼水面单位面积承受２２００ｋｇ／ｈａ网箱鱼的负载量设置网箱，建立以网箱养鱼为重要环节的水库新型渔业生产结构体系。三年中，水库水面鱼产量（含网箱鱼产量）由１４５．５ｋｇ／ｈａ上升到２６６．７１ｋｇ／ｈａ，年总产量增长１８．３倍；渔业投入资金年利润率达７１．９％；网箱养鱼的投入总能量转换效率提高了６．０３－６．３６％。合理开发和保护了水库渔业资源，促进了温杖子水库渔业的高速增产。