Lipid peroxidation,protein oxidant and antioxidant status of muscle and serum for juvenile Jian carp (Cyprinus carpio var. Jian) fed grade levels of methionine hydroxy analogue

Lipid peroxidation, protein oxidation and antioxidant status of serum and muscle in juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of methionine hydroxy analogue (MHA: 0, 5.1, 7.6, 10.2, 12.7, 15.3 g kg^?1 diet) for 60 days were investigated. Both malondialdehyde and protein carbonyl content in serum and muscle decreased with increasing dietary MHA level up to 5.1–10.2 g kg^?1 diet (P < 0.05). Anti‐hydroxyl radical and activities of catalase, glutathione peroxidase and glutathione reductase in muscle and serum, as well as anti‐superoxide anion, superoxide dismutase activity and glutathione content in serum, increased with optimal MHA supplement (P < 0.05). Meanwhile, glutathione‐S‐transferase activity in serum showed a downward trend with dietary MHA up to 7.6 g kg^?1 diet (P < 0.05). These results indicated that MHA improved antioxidant status and depressed lipid and protein oxidation in serum and muscle.     

Myo-inositol prevents oxidative damage, inhibits oxygen radical generation and increases antioxidant enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)

This study was conducted to evaluate the effects of dietary myo -inositol (MI) on the antioxidant status of juvenile Jian carp ( Cyprinus carpio var. Jian). A total of 1050 Jian carp (22.28±0.07 g) were randomly distributed into seven groups of three replicates each, feeding diets containing graded levels of MI (163.5, 232.7, 384.2, 535.8, 687.3, 838.8 and 990.3 mg kg⁻¹ diet) for 60 days. Results indicated that the malondialdehyde content was the lowest for fish fed diets containing ≥384.2 mg MI kg⁻¹, and the highest for fish fed the MI-unsupplemented basal diet ( P <0.05). The protein carbonyl content was decreased with increasing dietary MI levels up to 535.8 mg kg⁻¹ diet, and no differences were found with a further increase in the MI concentration. The anti-superoxide anion capacity (ASA) and anti-hydroxyl radical capacity (AHR) were increased with increasing MI levels up to 535.8 mg kg⁻¹ diet, and plateaued thereafter. The superoxide dismutase and glutathione- S -transferase activities showed the same tendency with the ASA capacity. Catalase, glutathione peroxidase and glutathione reducase activities were improved with increasing MI levels up to 838.8, 384.2 and 687.3 mg kg⁻¹ diet, respectively, and remained nearly constant thereafter. These results suggested that MI could inhibit oxygen radical generation, increase enzymatic antioxidant capacity and prevent oxidative damage of carp. Dietary MI requirements for ASA and AHR activities of juvenile Jian carp were 567.94 and 517.22 mg MI kg⁻¹ diet respectively.     

Influence of dietary zinc on lipid peroxidation,protein oxidation and antioxidant defence of juvenile Jian carp (Cyprinus carpio var. Jian)

This study was conducted to study the effects of dietary zinc on lipid peroxidation, protein oxidation and antioxidant defence of juvenile Jian carp (Cyprinus carpio var. Jian) by feeding fish with increasing levels of zinc (15.3, 26.9, 40.8, 58.2, 68.9 and 92.5 mg Zn kg^?1) for 6 weeks. Results indicated that malondialdehyde (MDA) content and protein carbonyls (PC) in serum were the highest in fish fed diet containing 15.3 mg zinc kg^?1 diet (P < 0.05). Serum antisuperoxide anion (ASA), superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px), glutathione reductase (GR) activities and glutathione (GSH) content were improved with increasing dietary zinc levels up to 40.8 mg zinc kg^?1 diet (P < 0.05) and levelled off (P > 0.05). Serum antihydroxy radical (AHR), catalase (CAT) and glutathione‐S‐transferase (GST) activities followed the similar pattern to that observed in ASA. The MDA and PC levels, ASA, AHR, SOD, CAT, GPx, GR, GST activities and GSH content in intestine, hepatosomatic and muscle tissue followed the similar pattern to that observed in serum. The present results indicated that zinc decreased lipid peroxidation and protein oxidation and improved antioxidant defence in fish.     

Effects of dietary methionine hydroxy analogue supplement on growth,protein deposition and intestinal enzymes activities of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 900 juvenile Jian carp (Cyprinus carpio var. Jian) (8.24 ± 0.03 g) were fed practical diets containing graded levels of methionine hydroxy analogue (MHA) (0, 5.1, 7.6, 10.2, 12.7 and 15.3 g kg^?1 diet) for 60 days to investigate the effects of MHA on growth, protein deposition and intestinal enzymes activities. Per cent weight gain (PWG) significantly increased with increasing levels of MHA up to a point (P < 0.05), and thereafter declined. Feed efficiency was the highest when MHA level was at 5.1 g kg^?1 diet. Feed intake (FI) significantly enhanced with dietary MHA level up to a point (P < 0.05), beyond which it plateaued. Patterns of differences in protein production value, lipid production value, intestosomatic index, folds height, lipase, chymotrypsin, γ‐glutamyl transpeptidase, Na⁺, K⁺‐ATPase, creatinekinase, glutamate‐oxaloacetate transaminase and glutamate‐pyruvate transaminase activities were the same as that in PWG, whereas the trend of plasma ammonia content was opposite. Hepatopancreas protein content, trypsin and amylase activities followed the same trend as that of FI. The optimal supplemental level of MHA for fish meal and miscellaneous meals–based diet (6.9 g methionine kg^?1 diet) for PWG was 8.2 g kg^?1 diet by the quadratic regression analysis.     

Dietary pyridoxine requirement of juvenile Jian carp (Cyprinus carpio var. Jian)

In a 80-day feeding trial, a total of 1050 juvenile Jian carp ( Cyprinus carpio var. Jian) with an average initial weight of 10.71 ± 0.05 g were fed semi-purified diets containing seven graded levels of pyridoxine (0.20, 1.71, 3.23, 4.96, 6.32, 8.58 and 12.39 mg pyridoxine kg⁻¹ diet). Results indicated that with increasing dietary pyridoxine levels up to 4.96 mg kg⁻¹ diet, percent weight gain (PWG) and specific growth rate (SGR) were improved, and no differences were found with further increase of pyridoxine levels. Feed intake also followed the similar pattern to that observed with PWG and SGR when dietary pyridoxine levels were ≤6.32 mg kg⁻¹ diet. But feed efficiency and protein efficiency ratio were not affected by pyridoxine levels. Crude protein of carcass, productive protein value and plasma ammonia concentration were improved with increasing dietary pyridoxine levels up to 4.96 mg kg⁻¹ diet. Amylase activities in the intestine were improved with increasing dietary pyridoxine levels up to 4.96 mg kg⁻¹ diet, but protease and lipase activities in the intestine were not affected by pyridoxine levels. Na⁺, K⁺-ATPase and Gamma-glutamyl transpeptidase activities in proximal intestine, mid intestine (MI) and distal intestine (DI) were lowest when fed the diet containing 1.71 mg pyridoxine kg⁻¹ diet. The alkaline phosphatase activities in MI and DI followed the same pattern. The dietary pyridoxine requirement of juvenile Jian carp based on PWG estimated by broken line model was 6.07 mg pyridoxine kg⁻¹ diet.     

Dietary phosphorus prevents oxidative damage and increases antioxidant enzyme activities in intestine and hepatopancreas of juvenile Jian carp

This study was conducted to investigate the effect of dietary phosphorus on the intestine and hepatopancreas antioxidant capacity of juvenile Jian carp (Cyprinus carpio var. Jian). Jian carp, with an average initial weight of 7.17 ± 0.01 g, were fed with diets containing graded concentrations of available phosphorus, namely 1.7 (control), 3.6, 5.5, 7.3, 9.2 and 11.0 g kg^?1 diet for 9 weeks. Results showed that, in intestine and hepatopancreas, content of malondialdehyde (MDA), protein carbonyl (PC) and glutathione (GSH), capacity of anti‐superoxide anion (ASA) and anti‐hydroxyl radical (AHR), and glutathione reductase (GR), catalase (CAT), glutathione S‐transferase (GST), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were significantly affected by dietary phosphorus levels (P < 0.05). Regression analysis showed that significant quadratic responses occurred in MDA content and ASA, GST, GPx and AHR activities in intestine, GSH content and CAT and SOD activities in hepatopancreas (P < 0.05). These results indicate that optimal level of dietary phosphorus prevented oxidative damage and increased antioxidant enzyme activities in the intestine and hepatopancreas of juvenile Jian carp. The phosphorus requirement estimated from MDA using quadratic regression analysis was 5.7 g kg^?1 diet.     

Effects of pyridoxine on antioxidative parameters in juvenile Jian carp (Cyprinus carpio var. Jian)

To investigate the effects of dietary pyridoxine (PN) on antioxidant status of fish in serum, intestine and trunk kidney, 1050 juvenile Jian carp (11.7 ± 0.1 g) were used for the experiment. The carp were divided into seven groups and fed diets containing graded levels of PN (0.20, 1.71, 3.23, 4.96, 6.32, 8.58 and 12.39 mg kg^?1 of diet) for 80 days. Results of the study showed that content of malondialdehyde in serum, intestine and kidney tissues was the highest when fed the diet containing 1.71 mg PN kg^?1 diet (P < 0.05). Meanwhile, the protein carbonyl content of intestine and kidney tissue showed a downward trend to a point (P < 0.05). Conversely, activities of superoxide dismutase, catalase, glutathione peroxidase (GSH‐Px), glutathione S‐transferase, glutathione reductase and glutathione (GSH) content in serum, intestine and kidney tissue were generally higher in PN‐supplemented diets than unsupplemented diet (P < 0.05). The present results indicated that the PN decreased lipid peroxidation and protein oxidation in fish, and partly because of its improved antioxidant enzymes activities and levels of GSH.     

Effects of dietary vitamin K levels on growth performance,enzyme activities and antioxidant status in the hepatopancreas and intestine of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 1400 juvenile Jian carp (Cyprinus carpio var. Jian) (8.93 ± 0.03 g) were fed diets containing graded levels of vitamin K at 0.027 (basal diet), 1.52, 3.02, 4.51, 6.02 and 7.52 mg kg^?1 diet for 60 days to investigate the effects of vitamin K on growth, enzyme activities and antioxidant capacity in the hepatopancreas and intestine. Percentage weight gain (PWG), feed intake and feed efficiency of fish were improved by vitamin K. Activities of trypsin, chymotrypsin, amylase and lipase in the intestine and hepatopancreas and Na⁺, K⁺‐ATPase, creatine kinase, alkaline phosphatase and gamma‐glutamyl transpeptidase in the intestine were increased by vitamin K. Malondialdehyde and protein carbonyl contents in the hepatopancreas and intestine were decreased with vitamin K supplements. Certain level of vitamin K increased antihydroxyl radical, antisuperoxide anion, superoxide dismutase, catalase, glutathione‐S‐transferase, glutathione peroxidase and glutathione reductase activities and glutathione contents in the hepatopancreas and intestine. Intestinal Lactobacillus, E. coli and Aeromonas were changed with vitamin K supplements. Together, these results indicate that vitamin K improved fish growth, digestive and absorptive ability, and anti‐oxidant capacity. The dietary vitamin K requirement of juvenile Jian carp (8.93–73.7 g) based on PWG was 3.13 mg kg^?1 diet.     

Dietary lysine requirement of juvenile Jian carp (cyprinus carpio var. Jian)

This study was designed to determine the dietary lysine requirement of juvenile Jian carp (cyprinus carpio var. Jian). A total of 990 juvenile Jian carps (initial weight 7.89 ± 0.04 g) were randomly allocated six groups of each three replicates, feeding isonitrogenous (320 g kg^?1 crude protein) and isolipidic (50 g kg^?1 crude fat) diets containing graded levels of coated l ‐lysine, with dietary lysine content ranging from 22 to 69 g kg^?1 of protein, respectively, for 80 days. Final weight, protein deposition and protein production value were significantly improved in response to the increasing levels of dietary lysine up to 59 g kg^?1 of protein and remained nearly constant thereafter. Whole‐body protein content followed a similar pattern as growth parameters in relation to dietary lysine level. Hepatopancreas and intestine protein content, hepatopancreas index (HSI) and intestine index (ISI) were positively affected by dietary lysine level. The dietary lysine requirement of juvenile Jian carp estimated by broken line analysis based on weight gain was 59 g kg^?1 of protein.     

Pb~(2+)对建鲤幼鱼抗氧化酶活性和总抗氧化能力的影响

为了解铅对建鲤幼鱼(Cyprinus carpio var.Jian)的影响,将建鲤幼鱼暴露于不同水平(0、0.648、1.296、6.480 mg/L)的Pb2+水溶液中,于96 h分别测定建鲤幼鱼肝胰脏、肠、鳃、肌肉中的超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活性及总抗氧化能力(T-AOC)。结果表明:随着Pb2+浓度的升高,抗氧化酶活性升高,达到极值后下降。0.648 mg/L浓度组中,肝胰脏中抗氧化酶活性及总抗氧化能力均有较明显升高;肌肉中抗氧化酶活性及总抗氧化能力变化较小;在高浓度组(6.480 mg/L),肾中的三种抗氧化酶活性及总抗氧化能力仍在升高,而在其他组织中酶的活性有所降低。上述研究结果表明,与肠、鳃、肌肉、肾相比,肝胰脏中的抗氧化酶活性及总抗氧化能力敏感,是最先启用的主要解毒器官;在高浓度胁迫时,肝胰脏解毒能力下降,抗氧化防御作用的主要器官由肝胰脏转移到肾脏。     

Effect of dietary thiamin supplement on immune responses and intestinal microflora in juvenile Jian carp (Cyprinus carpio var. Jian)

This study was to investigate the effect of dietary thiamin on the immune response and intestinal microflora in juvenile Jian carp (Cyprinus carpio var. Jian). Seven diets (0.25, 0.48, 0.79, 1.06, 1.37, 1.63 and 2.65 mg thiamin kg^?1) were fed to Jian carp (8.20 ± 0.02 g). At the end of feeding trial, red blood cell counts and white blood cell counts increased with increasing dietary thiamin levels up to 0.79 and 1.06 mg kg^?1 diet, respectively (P < 0.05); Lactobacillus counts in intestine increased gradually, and maximum values obtained when the thiamin level was at 0.79 mg kg^?1 diet (P < 0.05). After the feeding trial, fish were injected with Aeromonas hydrophila only one time on the first day of the challenge trial and fed the same diets as the growth trial for 17 days. The survival rate, leucocyte phagocytic activity, lectin potency, acid phosphatase activity, lysozyme activity, total iron‐binding capacity and immunoglobulin M content of fish after being injected with A. hydrophila were all improved with increasing dietary thiamin levels up to a point, respectively (P < 0.05). These results suggested that thiamin could improve immune response and the balance of intestinal microflora of juvenile Jian carp.     

Immune response,disease resistance and intestinal microflora of juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of pantothenic acid

This study was to investigate the effect of dietary pantothenic acid (PA) on the disease resistance, immune response and intestinal microflora on juvenile Jian carp (Cyprinus carpio var. Jian). Seven diets (4.0, 15.5, 25.6, 36.1, 45.9, 56.1 and 65.9 mg PA kg^?1) were fed to Jian carp (12.95 ± 0.03 g) for 9 weeks. After 9‐week feeding trial, the challenge experiment with Aeromonas hydrophila was conducted to determine the impact of PA on fish disease resistance. Survival rate after challenge was promoted with the increasing PA levels (P < 0.05). Blood counts also significantly increased up to the dietary PA level of 25.6 mg PA kg^?1 (P < 0.05). Leucocyte phagocytic activity, lectin potency, lysozyme and acid phosphatase activity, and total iron‐binding capacity were improved with increasing PA levels (P < 0.05). Serum immunoglobulin M level and agglutination antibody titre to A. hydrophila were increased (P < 0.05) in fish fed the diets with the dietary PA levels between 56.1 and 65.9 mg kg^?1. PA also promoted the growth and reproduction of Lactobacillus and depressed Escherichia coli and A. hydrophila (P < 0.05). These results suggested that pantothenic acid could improve disease resistance, immune response, and the balance of intestinal microflora in juvenile Jian carp.     

Effects of dietary histidine levels on growth performance,body composition and intestinal enzymes activities of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 1200 juvenile Jian carp (Cyprinus carpio var. Jian) (8.76 ± 0.02 g) were fed diets containing graded levels of histidine at 2.3 (unsupplemented control), 4.4, 6.3, 8.6, 10.8 and 12.7 g kg^?1 diet for 60 days to investigate the effects of histidine levels on growth performance, body composition, intestinal enzymes activities and microflora. Specific growth rate (SGR), feed efficiency, protein efficiency ratio, protein productive value, body protein content and lipid content of fish were lowest in fish fed the basal diet (P < 0.05). Activities of glutamate‐pyruvate transaminase in muscle and hepatopancreas, trypsin, chymotrypsin, amylase, lipase activities in intestine and hepatopancreas, and Na⁺, K⁺‐ATPase, creatine kinase, alkaline phosphatase, γ‐glutamyl transpeptidase activities in three intestinal segments were improved by dietary histidine (P < 0.05), whereas glutamate‐oxaloacetate transaminase activities and plasma ammonia content followed an opposite trend. The amounts of Lactobacillus, Escherichia coli and Aeromonas were significantly affected by dietary histidine levels (P < 0.05). These results suggested that histidine could improve growth and enhance intestinal enzymes activities of juvenile Jian carp. The dietary histidine requirement of juvenile Jian carp (8.76–68.02 g) based on SGR was 7.8 g kg^?1 diet or 2.38 g 100 g^?1 protein by quadratic regression analysis.     

维生素A缺乏对建鲤生长性能及免疫功能的影响

幼建鲤体重(11 37±0 55)g,投喂不添加或添加维生素A的3组试验饲料,进行饲养实验70d,攻毒实验15d和免疫接种实验24d,研究维生素A缺乏对幼建鲤生长性能和免疫功能的影响,结果表明,维生素A缺乏组(0IU/kg)建鲤的成活率、摄食量、增重、净蛋白质沉积率、肥满度、肝胰脏指数、肝胰脏维生素A含量、肠道皱襞高度,头肾重量、脾脏重量和指数、后肾重量和指数、血液红细胞、白细胞数量和血清溶菌酶活力显著降低(P<0 05),且饲料系数显著增加(P<0 05),而对头肾体指数、用灭活嗜水气单胞菌免疫后第10和17天血清抗体水平影响不显著(P>0 05)。基于本实验研究结果,可知维生素A是维持幼建鲤正常生长和免疫机能必不可少的营养成分。当饲料中维生素A含量达到3969IU/kg时,一定程度上可满足幼建鲤正常生长和免疫机能的需要,进一步提高饲料维生素A含量(达23816IU/kg),对幼建鲤生长和饲料利用没有显著影响,但在一定程度可提高其免疫机能。     

Use of supplemental amino acids on the success of reducing dietary protein levels for Jian carp (Cyprinus carpio var. Jian)

A feeding trial was conducted to investigate the effects of essential amino acids (EAA) supplementation on the success of dietary protein reduction for Jian carp (Cyprinus carpio var. Jian). Five diets were formulated including a positive control (Diet 1, 378.4 g/kg protein), a negative control (Diet 5, 289.8 g/kg protein) and three test diets (Diet 2‐Diet 4, 340.4–299.0 g/kg protein supplemented with EAA). No significant decline of growth performance in fish fed decreased dietary protein with EAA supplementation, which were significantly higher than the negative control. The apparent digestibility coefficients (ADC) of protein and most EAAs were not significantly affected by the dietary treatments. Dietary treatments did not affect whole‐body composition, but influenced protein and EAAs retentions. Total ammonia nitrogen (TAN) excretion decreased with the decreasing dietary protein, and the highest plasma ammonia content was observed in fish fed Diet 5. Plasma glucose concentration increased with the decreasing dietary protein reduction, while other plasma parameters were not affected by dietary treatments. These results indicated that low‐protein diets (299.0 g/kg) supplemented with EAA can maintain protein, EAA digestibility and retention in Jian carp, and also has benefit in the reduction of TAN excretion.     

Growth, digestive capacity and intestinal microflora of juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of dietary inositol

A 60‐day feeding trial was carried out with juvenile Jian carp (Cyprinus carpio var. Jian) to study the effects of myo‐inositol (MI) on the growth, digestive enzyme and intestinal microbial population. Diets with seven levels of inositol (163.5, 232.7, 384.2, 535.8, 687.3, 838.8 and 990.3 mg MI kg^?1 diet) were fed to Jian carp (initial weight 22.28±0.07 g). Per cent weight gain (PWG) was improved with increasing inositol levels up to 535.8 mg MI kg^?1 diet (P<0.05), and plateaued (P>0.05). The protein production value, lipid production value and ash production value were increased with increasing dietary inositol levels up to 384.2, 838.8 and 838.8 mg MI kg^?1 diet respectively (P<0.05). Although intestinal protein content and trypsin activity were not affected by inositol levels (P>0.05), chymotrypsin, lipase and amylase activities in intestine were the lowest for fish fed the MI‐unsupplemented diet (P<0.05). Alkaline phosphatase, Na⁺, K⁺‐ATPase, γ‐glutamyl transpeptidase and creatinkinase activities in the intestine were increased with an increase in the inositol levels up to 384.2–687.3 mg MI kg^?1 diet (P<0.05). Intestinal Aeromonas hydrophila and Escherichia coli decreased with an increase in the levels of dietary inositol up to 232.7 and 687.3 mg MI kg^?1 diet respectively (P<0.05), while Lactobacillus in the intestine increased with an increase in inositol levels up to 990.3 mg MI kg^?1 diet (P<0.05). In conclusion, inositol improved growth, digestive capacity and intestinal microbial population of juvenile Jian carp, and the dietary inositol requirement for PWG of juvenile Jian carp is 518.0 mg MI kg^?1 diet.     

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.     

油酸诱导建鲤(Cyprinus carpio var. Jian)肝细胞脂肪变性模型的建立

通过对油酸诱导时间及有效作用浓度的研究,建立油酸致建鲤肝细胞脂肪变性模型.本研究以胰蛋白酶消化法获得的原代肝细胞作为实验材料,加入不同浓度油酸(0、0.05、0.1、0.2、0.4、0.8 mmol/L)与肝细胞共培养24-48 h,诱导肝细胞脂肪变性,分别收集不同时段的肝细胞及上清液,采用试剂盒测定肝细胞中甘油三酯(TG)、总胆固醇(TC)的含量,同时测定上清培养液中谷丙转氨酶(GPT)、谷草转氨酶(GOT)、γ-谷氨酰转肽酶(γ-GT)、乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)的活性,MTT法测定肝细胞的存活情况,确定油酸最佳诱导浓度,油红O染色法观察细胞内脂肪滴的形成情况.研究结果显示,0.4 mmol/L油酸与肝细胞共培养48 h,可以显著提高肝细胞内TG、TC含量(P＜ 0.05或P＜ 0.01),极显著提高上清培养液中GOT、GPT、LDH、γ-GT的活性,显著降低上清培养液中SOD的活性,但与24 h相比有升高趋势,0.4 mmol/L油酸与肝细胞共培养24 h,光镜下可见细胞内有脂肪滴形成,以48 h最明显.综上所述,油酸浓度0.4 mmol/L、诱导时间48 h成功构建了肝细胞脂肪变性模型,为研究鱼类脂肪肝类疾病奠定了基础.     

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

A 9-week feeding trial was carried out with juvenile Jian carp to study the effect of dietary pantothenic acid (PA) on growth, body composition and intestinal enzyme activities. Semi-purified diets with seven levels (4.0, 15.5, 25.6, 36.1, 45.9, 56.1 and 65.9 mg PA kg⁻¹) of supplemental calcium d -pantothenate were fed to Jian carp (13.0 ± 0.0 g). PA improved specific growth rate (SGR), protein productive value (PPV), protein efficiency ratio (PER) and lipid production value (LPV) ( P <  0.05). Fish fed the control diet had significantly lower feed efficiency (FE) than that in any other group ( P <  0.05). Body protein content increased with increasing PA levels ( P <  0.05), but moisture, lipid and ash of fish carcasses were negatively related to the graded PA levels ( P <  0.05). Intestine protein content (IPC), hepatopancreas protein content (HPC) and activity of α-amylase, lipase, trypsin, Na⁺,K⁺-ATPase, alkaline phosphatase (AKP) and gamma-glutamyl transpeptidase (γ-GT) were all positively affected by the dietary PA levels ( P <  0.05), while intestine index (ISI) and hepatopancreas index (HSI) decreased with the increment of supplemental levels of PA ( P <  0.05). These results suggested that PA could enhance fish growth and intestinal enzyme activities. The dietary PA requirement of juvenile Jian carp, Cyprinus carpio var . Jian (13.0–73.0 g), for optimal growth estimated by the broken-line analysis was 23.0 mg PA kg⁻¹ diet.     

Effect of dietary iron levels on growth,body composition and intestinal enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)

A 60‐day feeding trial was carried out to investigate the effect of iron on growth, body composition and digestive enzyme activities. Diets with seven levels of iron (53.9, 90.0, 115.6, 146.1, 176.0, 215.8 and 266.0 mg iron kg^?1 diet) were fed to Jian carp (initial weight 11.4 ± 0.0 g). Per cent weight gain (PWG), feed efficiency (FE) and protein efficiency ratio were the lowest in fish fed the basal diet (P < 0.05). Body protein content was increased with the increasing iron levels (P < 0.05), but moisture, lipid and ash of fish were not significantly affected by dietary iron levels (P > 0.05). Activities of trypsin, lipase, α‐amylase, Na⁺, K⁺‐ATPase, alkaline phosphatase and gamma‐glutamyl transpeptidase were improved with increasing dietary iron levels. Serum iron were significantly enhanced with dietary iron levels up to 146.1 mg iron kg^?1 diet, and plateaued. In conclusion, iron improved digestive enzyme activities of juvenile Jian carp and the dietary iron requirement for serum iron of juvenile Jian carp (11.4–64.0 g) was 147.4 mg iron kg^?1 diet with ferrous fumarate as the iron source.