为建立高效液相色谱法测定甜叶菊提取物中绿原酸、新绿原酸、隐绿原酸、洋蓟素、异绿原酸B、异绿原酸A、异绿原酸C7种绿原酸及其类似物的分析方法,本实验以超声提取法对样品进行前处理,以甲酸溶液-乙腈作为流动相,采用梯度淋洗和C18色谱柱(250 mm×4.6 mm,5μm)进行各化合物分离.结果 显示:7种绿原酸及其类似物的检出限为0.47～1.18 mg/kg,相关系数r均大于0.9990,加标回收率为95.10％～102.63％,相对标准偏差(n=3)为1.7％～4.1％.将本方法应用于甜叶菊提取物中的绿原酸及其类似物的含量分析,实验结果满意.

烂鳃病,水产养殖最常见的疾病之一.按理养殖户对烂鳃病并不陌生,而且都能一眼就认出患病鱼是否患有烂鳃病.但是,通过笔者的走访发现,许多养殖户虽然能辨别鱼鳃是否溃烂,但用药后却常常治不住;或者以前治疗烂鳃非常有用的药,下一次用却没有明显的疗效,给养殖户造成巨大的损失.分析其原因,主要是养殖户只认得出烂鳃,却不能清楚的认识引起鱼烂鳃的主要发病诱因,因而造成下药不对症,浪费了治疗的宝贵时间,使病鱼继发其他新的病症.因此,本文将主要介绍烂鳃病的几种主要诱因及治疗方法,希望能使养殖户更清楚的区别和治疗烂鳃病.

在俄罗斯鲤基础饲料中分别添加0.2%的复合酸化剂和0.3%的复方中草药,研究复合酸化剂和复方中草药对鲤鱼生长和非特异性免疫性能的影响。结果表明:与对照组相比,添加复合酸化剂能显著提高鲤鱼的增重率、特定生长率和白细胞数量,显著降低饲料系数,但对采食量、血清谷草/谷丙转氨酶活性和红细胞数量无显著影响;与对照组相比,添加复方中草药对鲤鱼的增重率、特定生长率和饲料系数有一定的改善作用（P＞0.05）。添加复方中草药能显著降低血清谷草转氨酶活性,但对血清谷丙转氨酶活性及红细胞和白细胞数量无显著影响。结论:饲料添加0.2%的复合酸化剂能够促进鲤鱼生长,同时增强其非特异性免疫能力。添加0.3%的复方中草药能够有效保护鲤鱼肝胰。

This experiment was conducted to investigate the protective effect of different concentrations of vitamin C on hydrogen peroxide(H2O2)-induced oxidative damage in intestinal epithelial cells(IEC) of Jian carp.In 24-well culture plates,the Jian carp IEC were primarily cultured for 72 h,and then continued to culture in serum-free DMEM culture medium.After 12 h further culture,the IEC were randomly divided into 7 groups with 4 replicates per group.One of the 7 groups was set as blank control,and the other groups were all supplemented with 100 μmol/L H2O2 in culture medium in order to establish oxidative stress model.After 16 h further culture in the same condition,the culture medium was changed into the DMEM culture medium which contained 0(blank control group),0,4,10,16,22 and 28 mg/L vitamin C,respectively.The samples were collected to measure the indices after 72 h vitamin C culture.The results showed as follows: 1) the 100 μmol/L H2O2 treatment led to less survival number and small colony of IEC;significantly decreased the activities of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GPX),glutathione S-transferase(GST) and glutathion reductase(GR),and reduced the contents of glutathione(GSH) and protein(P0.05);significantly increased the contents of malonaldehyde(MDA) and protein carbonyl(PCA),and improved the lactate dehydrogenase(LDH) activity in culture medium(P0.05);caused oxidative stress of Jian carp IEC.2) Vitamin C significantly increased the anti-hydroxyl free radical ability(AHA) and anti-superoxide anion free radical ability(ASA)(P0.05),and recovered the oxidation damage in IEC of Jian carp induced by H2O2.In summary,vitamin C can improve the antioxidant ability of the Jian carp IEC,and effectively protect the IEC from oxidative damage.

This study was conducted to investigate the changes of activities of antioxidant enzymes,contents of non-enzymic antioxidant substances,as well as the anti-superoxide anion free radical ability(ASA) and anti-hydroxyl free radical ability(AHA) in juvenile Jian carp(Cyprinus carpio var.Jian) with different body weights(18 to 75 g).A total of 750 Jian carp with an average initial body weight of(18.0±0.2) g were randomly distributed into 5 groups with 3 replicates per group and 50 fish per replicate.The fish in the 5 groups were fed the same diets and were sampled every 14 days(total of 5 times).Briefly,after weighing,the blood,hepatopancreas,intestine and muscle were quickly collected for antioxidant indices analysis.The results showed as follows: contents of malondialdehyde(MDA) and protein carbonyls(PCA) in serum,hepatopancreas,intestine and muscle were significantly decreased with increasing body weight up to 31.4 and 58.6 g(P0.05),respectively,and no differences were found with further increase of body weight(P0.05).Adversely,ASA and AHA in serum and studied tissues were increased with increasing body weight up to 40.1 g(P0.05),and plateaued thereafter(P0.05).Interestingly,activities of superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GSH-Px) took the same trends with ASA.Similarly,activities of glutathione S-transferase(GST) and glutathione reductase(GR) in serum and tissues were all increased with increasing body weight up to 31.4 g(P0.05),and plateaued thereafter(P0.05).Furthermore,reduced glutathione(GSH) content in serum,hepatopancreas,intestine and muscle were also increased with increasing body weight up to 31.4,31.4,31.4 and 58.6 g(P0.05),respectively,and no changes were found with further increasing body weight(P0.05).Summarily,the results in this study indicate that in the juvenile period of Jian carp(18 to 75 g),the activities of SOD,CAT,GSH-Px,GST and GR,as well as GSH content in serum and tissues are increased with increasing body weight,and thus the ASA and AHA are also improved,and finally products of the lipid peroxidation and protein oxidation are decreased with increasing body weight until reaching steady state.

To investigate the effects of vitamin C on the proliferation,differentiation,structure,and function of carp intestinal epithelial cells in primary culture,the carp intestinal epithelial cells(IEC) in primary culture were cultured in 24-well plate after 72 h,then changed into the DMEM culture medium which contained 0,4,10,16,22,and 28 mg/L vitamin C,respectively.The samples were collected to measure the indices after 72 h vitamin C culture.The results showed that vitamin C deficiency lead to less IEC survival number and small colony;adding vitamin C,adherent cell number increased significantly,and IEC formatted the large colony and monolayer,and the MTT OD,alkaline phosphatase(AKP),alanine transaminase(GPT),glutamic-oxalacetic transaminase(GOT),Na+,K+-ATPase activities,and protein contents in IEC significantly improved(P0.05),lactate dehydrogenase(LDH) in culture medium reduced significantly(P0.05),although there were the differences among the diferent concentrations of vitamin C,eg.10 mg/L and 16 mg/L.In summary,vitamin C can promote proliferation and differentiation of carp IEC,improve the structure of the cell membrane integrity,enhance intestinal cell function and metabolism of amino acids,increase cell protein deposition.

The experiment was conducted to study the effects of different sources and levels of vitamin C on growth performance and digestive function of Jian carp(Cyprinus carpio var.Jian).Six hundred Jian carp with initial average body weight of(12.63±0.04) g were randomly allocated to 4 groups(with 3 replicates per group and 50 fish per replicate).The Jian carp were fed with practice diets supplemented with different levels(75 or 150 mg/kg diet) of ethylcellulose-coated ascorbic acid(EC-AA) or L-ascorbyl polyphosphate(LAPP) for 8 weeks.When supplemented with 75 mg/kg vitamin C in diets,specific growth rate(SGR),feed intake(FI),protein production value(PPV) and lipid production value(LPV) of carp in LAPP group were all significantly higher than those in EC-AA group(P0.05).Meanwhile,compared with the EC-AA diet,the LAPP diet could significantly improve the growth and development of hepatopancreas and intestine,and enhance the activities of tryspin,chymotrypsin and lipase in intestine,folds height in different intestine segments,activities of alkaline phosphatase(AKP) and glutamyl transpeptidase(γ-GT) in different intestine segments,and activities of Na+,K+-ATPase in proximal and distal intestine segments and creatine kinase(CK) in whole intestine(P0.05).Whereas,when supplemented with 150 mg/kg vitamin C in diets,the SGR,FI,PPV and LPV were not significantly different between LAPP group and EC-AA group(P0.05).However,compared with the EC-AA diet,the LAPP diet could significantly enhance the activities of tryspin and chymotrypsin in intestine,folds height in mid and distal intestine segments,activities of AKP in distal intestine,Na+,K+-ATPase in proximal intestine and γ-GT in different intestine segments(P0.05).In conclusion,the effect of LAPP is better than EC-AA for Jian carp,and the LAPP supplementation of 75 mg/kg vitamin C can more significantly improve the growth performance and digestion function of Jian carp.

A 60-day experiment was carried out to study the effects of vitamin C [ascorbic acid (AA)] on the growth, digestive enzyme activities and intestinal microbial population. Diets with six levels (0.0, 21.4, 45.1, 69.5, 93.6 and 142.1 mg AA kg-1 diet) of supplemental ascorbyl polyphosphate were fed to juvenile Jian carp (Cyprinus carpio var. Jian) (12.63 +/- 0.02 g). The specific growth rate (SGR), feed efficiency and productive protein value were significantly improved with increasing AA levels up to 45.1 mg AA kg-1 diet (P < 0.05). The hepatopancreas and muscle AA concentrations were increased with increasing dietary AA levels up to 69.5 and 45.1 mg kg-1 diet respectively (P < 0.05). The activities of intestinal trypsin, chymotrypsin, lipase, alpha-amylase, Na+, K+-ATPase, alkaline phosphatase, gamma-glutamyl transpeptidase and creatinkinase were all positively affected by the AA supplementation (P < 0.05). Intestinal Lactobacillus and Bacillus were increased with increasing AA levels up to 45.1 mg AA kg-1 diet (P < 0.05), while intestinal Escherichia coli decreased with increasing AA levels up to 45.1 mg AA kg-1 diet (P < 0.05). In conclusion, AA improve the digestive capacity and intestinal microbial population of Jian carp, and the dietary AA requirement for SGR of juvenile Jian carp was 40.9 mg AA kg-1 diet.

<正>通常,抗菌肽是动物免疫防御系统在外界条件刺激下,由基因编码、核糖体合成,用以对抗外源性病原体的一类含有20～60个氨基酸残基的多肽类物质。其具有快速、广谱、非特异性的抗菌特性,以及部

A total 192 healthy Cyprinus carpiovar.Jian carps(body weight 11.18 g±0.57 g,P0.05) were divided randomly into groups A and B.Group A was fed with the semi-purified diet supplemented with 60 mg vitamin C per kg as control and Group B was fed with the experimental diet without vitamin C for 133 days.The histopathological observation was carried out.In Group A,hepatocytes were degenerated and necrotic,renal tubular epithelial cells were degenerated,necrotic and exfoliated,splenic reticular stromal cells were swelled,myocardial fiber were swelled and degenerated,and muscle fiber were degenerated and necrotic.Under electronic microscopy,hepatin and enzyme particles were dissolved and looked like cavitation in pancreas cell,muscular fibrocyte and intestinal epithelium villi were ruptured,and splenic reticular stromal cells were swelled.The results revealed that different histopathological injuries were caused by vitamin C deficiency in different tissues and organs of juvenile C.carpio var.Jian.