Effects of ferulic acid on growth performance,immunity and antioxidant status in genetically improved farmed tilapia (Oreochromis niloticus) fed oxidized fish oil

A study was conducted to characterize the effects of dietary oxidized fish oil on the growth performance, immunity and antioxidant status of genetically improved farmed tilapia (Oreochromis niloticus) and to determine the role of ferulic acid on the oxidative damage induced by the oxidized fish oil. The tilapia (13.73 ± 0.31 g) were fed four experimental diets containing untreated (peroxide value, POV: 2.2 meq/kg) and highly oxidized (POV: 120.6 meq/kg) fish oil either with or without ferulic acid (0 or 400 mg/kg) supplementation for 12 weeks. From the results, the oxidized fish oil treatments increased antioxidant enzyme activities and MDA values but decreased the weight gain and the immunological parameters in tilapia. Meanwhile, the serum biochemical indices were significantly affected by the oxidized fish oil. Besides, the addition of ferulic acid partially counteracted the free radical‐induced damage and improved the health status of tilapia. In conclusion, the oxidized fish oil may induce oxidative stress, destroy liver, dysregulate lipid metabolism as well as reduce non‐specific immunity, and eventually result in growth inhibition of tilapia. The ferulic acid supplementation partially offset the negative effects of the oxidized fish oil on tilapia.     

车前子提取物抑制扑热息痛诱导小鼠肝损伤的作用研究

【目的】研究车前子提取物干预扑热息痛(Acetaminophen,APAP)引起小鼠肝损伤的作用,并探究其潜在作用机制。【方法】运用半仿生酶法提取车前子活性成分,采用液相色谱-质谱联用(LC-MS)检测其主要成分。体内研究车前子提取物对APAP肝损伤的保护作用,将60只雌性昆明小白鼠随机分为正常对照组(NC)、APAP肝损伤模型组(LD)、车前子对照组(PA)以及APAP干预的车前子提取物高、中、低剂量组(HPA+LC、MPA+LD、LPA+LD)。其中NC组和LD组小鼠灌胃生理盐水、PA组小鼠灌胃200μg/mL车前子提取物,HPA+LD,MPA+LD和LPA+LD组小鼠分别灌胃车前子提取物200,100,50μg/mL,每天2次,连续6 d;末次给药12 h后,采血并分离肝脏,检测血清谷丙转氨酶(ALT)和谷草转氨酶(AST)活性,评价肝脏损伤程度;检测肝脏中超氧化物歧化酶(SOD)活性及谷胱甘肽(GSH)、过氧化氢(H_2O_2)和丙二醛(MDA)含量,评价肝脏的氧化应激程度;检测CYP2E1 mRNA和蛋白表达,并通过细胞色素P450酶2E1(Cytochrome 2E1,CYP2E1)法体外筛选车前子抗APAP肝损伤可能的有效成分。【结果】LC-MS检测发现,车前子提取物有37种主要成分;血清生化指标检测发现,不同剂量的车前子提取物明显减少APAP诱导的氧化产物,增强抗氧化防御能力,减轻肝脏损伤且呈剂量依赖性;车前子提取物显著降低CYP2E1 mRNA和蛋白表达,说明车前子抑制CYP2E1的活性;车前子提取物中抗APAP肝损伤的有效成分为车前草苷D、车前草苷E、高车前素和高车前苷。【结论】车前子提取物的有效成分为车前草苷D、车前草苷E、高车前素和高车前苷。车前子提取物对APAP诱导的肝损伤具有保护作用,其潜在作用机制可能与抑制肝脏CYP2E1的表达和活性有关。     

Blood and liver biopsy for the non‐destructive screening of tilapia lake virus

Detection of tilapia lake virus (TiLV) in tilapines is mainly from visceral organs of killed fish. However, lethal sampling might not be viable to broodstock and economically important ornamental cichlids. To contribute towards screening of the virus in asymptomatic infected fish, a subclinically infected population of Nile tilapia adults obtained from a local farm was preliminarily tested to compare different non‐lethal sampling methods, for example liver biopsy, gill biopsy, fin clip, mucus, faeces and blood for detection of TiLV. Only liver and blood samples gave positive results by PCR. Since blood sampling is relatively simpler, it was further used for five naturally co‐cultured juvenile fish species from above‐mentioned farm including 40 red tilapia broodstock and 20 Nile tilapia adults from two other different farms. The results showed that from the tested fish, 4 of 5 Nile tilapia, 2 of 5 hybrid red tilapia and 3 of 5 giant gourami blood samples tested positive, while 38 of 40 blood samples of red tilapia tested positive for TiLV in second‐step PCR. Sequencing representative PCR amplicons of positive samples confirmed sequence identity to TiLV. In conclusion, both blood and liver biopsy are practical non‐destructive sampling platforms for TiLV screening in cichlids with blood being more convenient, especially for tilapia broodstock.     

Identification and establishment of sorting method for isolating CD11b+ myeloid cells in human hepatic metastases from colorectal cancer

AIM: To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b⁺ myeloid cells in hepatic metastases from colorectal cancer.METHODS: Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion, and then the CD11b⁺ myeloid cells were isolated by flow cytometry. The sorted cells were identified by immunocytochemistry. The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining. The cell purity was evaluated by flow cytometry.RESULTS: Sufficient numbers of CD11b⁺ cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion. The purity of the cells was confirmed by statistical analysis (P<0.05). The positive rates of the cells before and after sorting were significantly different (P<0.01). The positive cells were verified by immunocytochemical method. Meanwhile, the sorted cells had complete morphology and good activity.CONCLUSION: The CD11b⁺ myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity, good activity and complete morphology.     

Do plant‐based diets for Atlantic cod (Gadus morhua L.) need additions of crystalline lysine or methionine?

Atlantic cod (Gadus morhua), initial weight 15 g, were fed ten experimental diets for 15 weeks. The diets were based on a mixture of plant proteins (PP) and fish meal (FM), where PP constituted 65% of dietary protein. PP mixtures were chosen to reach as low levels of lysine and methionine as possible. The diets were supplemented with increasing amounts of lysine (19.2–31.9 g kg^?1 diet) or methionine (9.4–12.3 g kg^?1 diet), in a regression design. No growth difference among diet groups was found in the plant‐based diets. Increased dietary lysine resulted in decreased liver size, plasma triacylglycerol concentration (TAG) and lipid productive value (LPV). Methionine additions did not result in changed Hepatosomatic index (HSI), LPV or plasma TAG. Feed conversion ratio (FCR) and protein utilization were neither affected by lysine nor methionine. Plasma and muscle concentrations of free lysine and methionine correlated with dietary levels 5‐h post feeding. Overall conclusion was that cod maintain growth rates in plant‐based diets if dietary protein was high, without additional supplements of crystalline lysine or methionine. Lysine intake significantly influenced lipid metabolism, showing the necessity to add lysine in plant protein‐based diets to hinder increased lipid deposition. No such effects were found because of lack of methionine additions.     

Study on Developmental Expression of ApoCⅡ Gene mRNA in Liver Tissues of Pigs

The aim of this study was to investigate the developmental patterns of ApoCⅡ gene mRNA in liver in Mashen and Large White pigs, and study the relationship between the expression level of ApoCⅡ and the lipid metabolism in pigs.The mRNA relative expressions of ApoCⅡ gene in liver at seven stages of 1,30,60,90,120,150,and 180-day old in Mashen and Large White pigs were determined by quantitative Real-time PCR.The results showed that the developmental trend of ApoCⅡ mRNA expression in liver between Mashen and Large White pigs was different.The ApoCⅡ mRNA abundance was decreased from birth to 60-day old,then increased at 90-day old,and decreased again after that in Mashen pig.However,the relative expression amount in Large White pig was gradually decreased from birth to 150-day old and increased again at 180-day old.Except for the ApoCⅡ mRNA expression amount at 1-day old,the differences of the expression amount at other stages in Mashen and Large White pigs were significant or extremely significant (P<0.05 or P<0.01).The ApoCⅡ mRNA expression in liver was affected by age and breed,and could play an important role in lipid metabolism in pigs.     

代谢组学在反刍动物营养研究中的应用进展

代谢组学是研究生物系统中所有代谢物的一门新兴学科。它能够检测生命系统中某一特定生物层次的所有代谢物的变化,并能够利用其特有的分析技术平台和数据分析平台对所得代谢物进行系统性度量。近年来,因其测试技术的广泛适用性,代谢组学逐渐被应用于反刍动物的营养学研究中。本文从瘤胃代谢组学、肝脏代谢组学、乳腺代谢组学和血液代谢组学等4个方面对代谢组学在反刍动物营养研究中的应用进展进行了综述。     

Proteomics analysis of mitochondrial extract from liver of female zebrafish undergoing starvation and refeeding

Many fish species can withstand long period of food deprivation and consequently compensate for any weight loss by undergoing rapid growth during resumption of feeding. There is an interest in taking advantage of compensatory growth to reduce feed cost. In this present study, we attempted to compare the changes in hepatic mitochondrial proteome of zebrafish undergoing starvation and refeeding. Two‐dimensional gel separation and image analysis revealed a total of 65 spots that showed changes in expression after 15 days of starvation followed by 7 days of refeeding. A total of 35 proteins were selected for mass spectrometry analysis, resulting in the positive identification of 18 proteins. Identified proteins indicated that starvation resulted in reduction in glycolysis and increase in gluconeogenesis, while refeeding caused these activities to return to normal levels. Expression pattern of several proteins related to fatty acid and amino acid metabolism also suggested the utilization of non‐carbohydrate resources for energy during starving conditions. Proteins with chaperoning and antioxidative roles such as glucose‐regulated protein, paraxonase and heat‐shock protein were also upregulated in starved conditions.     

The effect of temperature and dietary fat level on tissue lipid composition in Atlantic salmon (Salmo salar) fed wax ester‐rich oil from Calanus finmarchicus

Copepod oil (CO) from the marine zooplankton, Calanus finmarchicus, is a potential alternative to fish oils (FOs) for inclusion in aquafeeds. The oil is composed mainly of wax esters (WE) containing high levels of saturated fatty acids (SFAs) and monounsaturated fatty alcohols that are poorly digested by fish at low temperatures. Consequently, tissue lipid compositions may be adversely affected in salmon‐fed CO at low temperatures. This study examined the lipid and FA compositions of muscle and liver of Atlantic salmon reared at two temperatures (3 and 12 °C) and fed diets containing either FO or CO, supplying 50% of dietary lipid as WE, at two fat levels (～330 g kg^?1, high; ～180 g kg^?1, low). Fish were acclimatized to rearing temperature for 1 month and then fed one of four diets: high‐fat fish oil (HFFO), high‐fat Calanus oil (HFCO), low‐fat fish oil (LFFO) and low‐fat Calanus oil (LFCO). The fish were grown to produce an approximate doubling of initial weight at harvest (220 days at 3 °C and 67 days at 12 °C), and lipid content, lipid class composition and FA composition of liver and muscle were determined. The differences in tissue lipid composition between dietary groups were relatively small. The majority of FA in triacylglycerols (TAG) in both tissues were monounsaturated, and their levels were generally higher at 3 °C than 12 °C. Polyunsaturated fatty acids (PUFA), particularly (n‐3) PUFA, predominated in the polar lipids, and their level was not significantly affected by temperature. The PUFA content of TAG was highest (～26%) in the muscle of fish fed the HFCO diet at both temperatures. Tissue levels of SFAs were lower in fish‐fed diets containing HFCO than those fed HFFO, LFFO or LFCO, particularly at 3 °C. The results are consistent with Atlantic salmon being able to incorporate both the FA and fatty alcohol components of WE into tissue lipids but, overall, the effects of environmental temperature on tissue lipids were more pronounced in fish fed the CO diets than FO diets.