Effects of in vitro exposure to ozone and/or hyperoxia on superoxide dismutase, catalase, glutathione and lipid peroxidation in red blood cells and plasma of rainbow trout, Oncorhynchus mykiss (Walbaum)

In aquaculture, ozone is used as a disinfectant. In its production, extensive amounts of oxygen are formed resulting in hyperoxic conditions in culture units. Both ozone and hyperoxia have the potential to be toxic via pro‐oxidant mechanisms and to activate antioxidant defence systems in cultured species. To eliminate systemic effects, blood of rainbow trout, Oncorhynchus mykiss (Walbaum), was exposed in vitro for 5 min to ozone/hyperoxia or hyperoxia, and changes in antioxidant defences and lipid peroxidation were measured after exposure. Ozone exposure caused severe damage in red blood cells (rbc) detected as increased lipid peroxidation and oxidized glutathione (GSSG) levels in both plasma and rbc. Oxygen exposure alone increased intracellular lipid peroxidation and GSSG levels 10 min after exposure and was not evident in the plasma at any time. Ozone, but not oxygen exposure, decreased reduced glutathione (GSH) levels in plasma, and the changes were negatively correlated with increased lipid peroxidation in rbc, indicating that extracellular GSH has a dynamic role in the protection of rbc from direct oxidation by ozone. Both ozone and hyperoxic conditions increased superoxide dismutase (SOD) activity in rbc 3 and 6 h after exposure. In contrast, catalase activity was only increased 10 min after oxygen exposure, suggesting other catalase activation mechanisms rather than enzyme induction. The recovery of lipid peroxidation and GSSG levels in rbc after hyperoxia, but not ozone exposure, indicated a capacity to defend against hyperoxia‐produced oxidative damage, but an overwhelming of antioxidant defences by ozone in rainbow trout rbc in vitro.     

Digestive enzyme activity of the red porgy (Pagrus pagrus, L.) during larval development under culture conditions

The ontogenic development of the main digestive enzymes (proteases, amylase and lipase) in the red porgy, Pagrus pagrus, larvae was assayed during the larval development. The green water technique was carried out for larval rearing and whole‐body homogenates were used for enzymatic assays in triplicate. Significant alterations in specific activities of all digestive enzymes measured during the period of this study were mostly related to metamorphosis and weaning. Trypsin‐ and chymotrypsin‐specific activities were first detected on day 3, together with opening of the mouth, and slightly increased until 25 days after hatching (DAH). After this period, the specific activities of these enzymes slightly decreased. Pepsin was first detected on day 28, concurrent with stomach formation, and a sharp increase was observed until 30 DAH. A slight decrease was measured from this date until the end of the experiment. Both amylase and lipase were measured for the first time on days 2 and 4 respectively, and the specific activities of these enzymes showed similar patterns during the first week of the study. Then, slight variations were observed until 30 DAH and while lipase‐specific activity declined, an increase in the specific activity of amylase was found until the end of the experiment. It is concluded that the variations observed in the specific activity of digestive enzymes were related to either metamorphosis, such as the formation of the stomach (28 DAH), or to changes in food composition. The profile of the developmental pattern of the main digestive enzymes detected in P. pagrus is similar to that described for other Sparid species.     

Effects of illumination on early life development and digestive enzyme activities in common pandora Pagellus erythrinus L. larvae

The activities of main digestive enzyme (proteases, amylase and lipase) and animal husbandry (mainly growth and survival) were studied in common pandora Pagellus erythrinus larvae until 30 days after hatching (DAH). Three different illumination levels (10, 30 and 100 lx) were compared in triplicate and green water technique was carried out. At the end of the experiment, illumination did not affect neither survival nor growth except 10 lx treatment. Similarly, specific growth rate (SGR) was different in 10 lx treatment (p < 0.05), although no differences were found in group 30 and 100 lx treatments (p > 0.05). In all groups, trypsin and chymotrypsin specific activities were firstly detected on day 3 related with mouth opening and slightly increased until 20 and 25 DAH respectively, and after this date specific activities of those decreased. Although, there was a significant difference between 10 lx treatment and other experimental groups (p < 0.05), no differences were found in other treatments (p > 0.05). Pepsin was firstly detected on day 25 related with stomach formation and sharp increase was observed until 30 DAH and then slight decrease was measured from this date and no differences were found between all groups. Amylase was firstly determined on day 2 and increased to day 5. After this date, slight decreases were measured in all groups and continued until end of experiments. The highest specific activity of amylase was determined in 30 lx treatment and no significant differences were found between groups (p > 0.05). Lipase was firstly detected on day 4 and increased to day 10. Then, activity of lipase decreased until day 15 and increased again until 25 DAH. Slight decreases were found in all groups until day 30 and continued to end of experiments. No significant differences were detected among groups (p > 0.05).

Finally, the significant improvement in survival, larval development and specific enzyme activities of larvae were determined in 30 lx treated group. It is thought that this phenomenon is related to optimal keeping conditions provided by the medium illumination level for Pagellus erythrinus larvae.     

Stimulating effects of pond water on digestive enzyme activity in the Pacific white shrimp, Litopenaeus vannamei (Boone)

Crude enzyme extracts were obtained from the digestive glands of Pacific white shrimp, Litopenaeus vannamei (Boone), reared in oligotrophic well water and eutrophic shrimp pond water to compare digestive enzyme activity between the two groups. Specific activities of serine protease, collagenase, amylase, cellulase, lipase and acid phosphatase were significantly higher (P < 0.01) in pond water‐reared shrimp (PW shrimp) than in well water reared‐shrimp (WW shrimp). For most enzymes assayed, specific activity was more than two times higher in PW shrimp, and cellulase activity was over six times higher. In contrast, chitinase activity was significantly higher (P < 0.001) in WW shrimp. Higher specific activity of most digestive enzymes in PW shrimp was probably due to natural productivity in the pond water that served as a source of organic substrates, and this increased activity may contribute to the growth‐enhancing effect of shrimp pond water.     

Effect of dietary carbohydrate-to-lipid ratios on growth, lipid deposition and metabolic hepatic enzymes in juvenile Senegalese sole (Solea senegalensis, Kaup)

A study was undertaken to determine the effect of various dietary carbohydrate‐to‐lipid ratios on growth performance, whole‐body composition and tissue lipid content in Senegalese sole (Solea senegalensis) juveniles. Data on the dietary regulation of key hepatic enzymes of the lipogenic and glycolytic pathways (glucose‐6‐phosphate dehydrogenase, G6PD; malic enzyme, ME; fatty acid synthetase, FAS; pyruvate kinase, PK and glucokinase, GK) were also generated. Four isonitrogenous (crude protein: 52% dry matter (DM)) diets were formulated to contain one of two lipid levels (11% and 21% DM). Within each dietary lipid level, the nature of the carbohydrate fraction (raw or extruded peas) was varied. Triplicate groups of 54 sole (initial body weight: 23.6±1.2 g) were grown in recirculated seawater over 67 days. Fish were fed using automated feeders. At the end of the study, whole‐body, liver, viscera and muscle samples were withdrawn for analyses. During the experimental period, the mean fish weight about doubled in all treatments. No significant differences were found in growth performance (ranging from 1.1% to 1.4% body weight day^?1) among dietary treatments. High‐fat diets increased whole‐body fat content. Similarly, daily fat gain ranged from 0.54 to 0.78 g kg^?1 day^?1 and highest values were found in fish fed high‐lipid diets. Dietary treatments also affected tissue lipid content (liver, viscera and muscle), with highest values in fish fed high‐fat diets. The nature of dietary carbohydrates had little influence on performance criteria, but affected tissue lipid deposition. The activities of G6PD, ME and FAS were depressed by elevated levels of dietary lipid, confirming the inhibitory effect of dietary fats on lipid biosynthesis. At both dietary lipid levels, ME and FAS activities were little affected by dietary carbohydrate. Activities of PK and GK were not affected by the starch level of the diets. In Senegalese sole juveniles, the lipogenic pathway is more susceptible to modulation by dietary means (particularly through lipid intake) than the glycolytic pathway.     

Trypsin enzyme activity during larval development of Litopenaeus vannamei (Boone) fed on live feeds

Larval stages of the Pacific white shrimp, Litopenaeus vannamei (Boone) were fed standard live diets of mixed microalgae from the first to the third protozoea (PZ1 to PZ3), followed by Artemia nauplii until post‐larvae 1 (PL1). Trypsin enzyme activity for each larval stage was determined using N‐α‐p‐toluenesulphonyl‐l ‐arginine methyl ester (TAME) as a substrate. Results were expressed as enzyme content to assess ontogenetic changes during larval development. Tissue trypsin content (IU µg^?1 DW for each larval stage) was significantly highest at the PZ1 stage and declined through subsequent stages to PL1. This contrasts with previously observed patterns of trypsin development in Litopenaeus setiferus (Linnaeus) and other penaeid genera, which exhibit a peak in trypsin activity at the third protozoea/first mysis (PZ3/M1) larval stage. Litopenaeus vannamei larvae transferred to a diet of Artemia at the beginning of the second protozoea (PZ2) stage were significantly heavier on reaching the first mysis stage (M1) than those fed algae, while survival was not significantly different between treatments. At both PZ2 and PZ3 stages, trypsin content in larvae feeding on Artemia was significantly lower than in those feeding on algae. The rapid decline in trypsin content from PZ1 and the flexible enzyme response from PZ2 suggest that L. vannamei is physiologically adapted to transfer to a more carnivorous diet during the mid‐protozoeal stages.     

橡胶树体胚发生过程中的生理生化特性

以巴西橡胶树花药为外植体,研究了巴西橡胶树花药体胚发生过程中抗氧化酶活性和某些生理参数的变化。结果表明,橡胶树花药体胚发生过程中,SOD活性变化表现在培养30 d达到最高值,而后下降平缓;丙二醛(MDA)含量在第60天达到最高。生理参数的变化表现为可溶性蛋白含量0 d含量最高,可溶性糖的含量在50 d达到最高,据此认为,SOD和MDA对橡胶树花药体胚的诱导起主导作用,可溶性蛋白含量与不定芽的发生密切相关。     

5种细胞质雄性不育小麦败育过程中活性氧代谢保护酶的响应

为了解异质雄性不育小麦败育过程中活性氧代谢中几种保护酶的变化特点,以5种小麦雄性不育系U116A、Ju116A、Va116A、C6116A、K116A及其相应保持系116B为材料,分析了小麦雄性不育系在花药不同发育时期的过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性与保持系的差异。结果表明,不育花药的POD活性自单核早期后均比同一发育时期的可育花药高;自单核晚期开始,不育系CAT活性总体上呈下降趋势,而可育系则呈升高趋势;受试材料花药中SOD活性总体呈上升趋势,且不育系高于保持系。经差异分析,不育系U116A、Ju116A、Va116A、C6116A的败育关键期是单核晚期,而K116A的败育关键期则是单核晚期至二核期。以上结果说明,5种异质小麦雄性败育时期有所不同,并且雄性育性与POD、CAT和SOD活性密切相关。     

控释肥配施玉米秸秆对麦季土壤酶活性及养分的影响

为了探究控释尿素掺混肥与玉米秸秆长期互作对麦季土壤酶活性和土壤养分状况的影响,基于华北平原棕壤区小麦—玉米轮作8年定位施肥试验,对比研究了玉米秸秆还田(S)与不还田条件下施用控释尿素掺混肥(CRF)与普通尿素掺混肥(BBF)对麦季土壤酶活性及土壤养分的影响。结果表明:秸秆不还田条件下,CRF处理较BBF处理显著提高土壤中性磷酸酶(返青期和成熟期)、蔗糖酶(返青期和孕穗期)和纤维素酶(返青期和孕穗期)活性及成熟期有机质、硝态氮和有效磷含量,其中,成熟期中性磷酸酶活性显著提高29.6%,硝态氮含量显著提高34.8%。秸秆还田条件下,BBF+S处理成熟期脲酶、纤维素酶,孕穗期中性磷酸酶活性显著高于CRF+S处理,其他时期2种类型掺混肥土壤酶活性无显著差异。CRF+S处理成熟期土壤全氮、有机质、硝态氮和有效磷含量显著高于BBF+S处理。氮肥种类和玉米秸秆还田的交互作用对土壤中性磷酸酶、蔗糖酶和纤维素酶活性及土壤全氮、硝态氮、铵态氮、有效磷和速效钾含量影响显著,秸秆还田较不还田处理显著提高中性磷酸酶、蔗糖酶活性、速效钾和有效磷含量。综上表明,玉米秸秆还田和不还田条件下控释尿素掺混肥较普通尿素掺混肥处理均能显著增加土壤有机质含量,提高小麦后期速效氮磷养分的供应强度和供应容量,其中玉米秸秆还田配施控释尿素掺混肥处理在提高土壤酶活性和土壤养分方面表现最优。研究结果可为秸秆还田下氮肥优化施用提供理论依据。