Challenge with 17α-ethinylestradiol (EE2) during early development persistently impairs growth,differentiation, and local expression of IGF-I and IGF-II in immune organs of tilapia

The enormous expansion of world-wide aquaculture has led to increasing interest in the regulation of fish immune system. Estrogen has recently been shown to inhibit the endocrine (liver-derived) and autocrine/paracrine local insulin-like growth factor-I system in fish. In order to address the potential actions of estrogen on the IGF system in immune organs, tilapia were fed with 17α-ethinylestradiol (EE2)-enriched food from 10 to 40 days post fertilization (DPF) to induce functional feminization, an approach commonly used in aquaculture. EE2-treated and control fish were sampled at 75 and 165 DPF. The expression levels of ER-α, IGF-I, IGF-II and growth hormone receptor (GH-R) mRNA in spleen and head kidney were determined by real-time PCR and the expressing sites of IGF-I mRNA identified by in situ hybridisation. Ratios of spleen length and weight to body length and weight were determined. At 165 DPF, the length (4.9% vs. 7.6%) and weight (0.084% vs. 0.132%) ratios were significantly lowered in EE2-treated fish and number and size of the melanomacrophage centres were considerably reduced. At 75 DPF, both in spleen and head kidney of EE2-treated fish the expression levels of IGF-I and IGF-II mRNA were markedly diminished. The suppression was more pronounced for IGF-I (spleen: ?12.071-fold; head kidney: ?8.413-fold) than for IGF-II (spleen: ?4.102-fold; head kidney: ?1.342-fold). In agreement, clearly fewer leucocytes and macrophages in head kidney and spleen of EE2-treated fish contained IGF-I mRNA as shown by in situ hybridisation. ER-α mRNA expression in spleen was increased at 75 DPF but unchanged in head kidney. GH-R gene expression showed a mild upregulation at 165 DPF in both tissues. Thus, exposure to EE2 during early development affected distinctly the IGF system in tilapia immune organs. It led to lasting impairment of spleen growth and differentiation that can be attributed to an interaction of EE2 with IGF-I and, less pronouncedly, IGF-II. Especially, the impairment of spleen and melanomacrophage centres might interfere with the antigen presentation capacity of the immune system and, thus, alter susceptibility to infection.     

The effects of splenectomy and subsequent starvation on the storage of haemosiderin by the melano-macrophages of rainbow trout Salmo gairdneri Richardson

In clinically normal, cachectic and diseased teleost fish, haemosiderin is observed mostly in the melano-macrophage centres of the spleen and to a much lesser extent in the centres of the kidney and liver. When rainbow trout that had been splenectomised were starved, accumulation of haemosiderin was diverted to the kidney melano-macrophages.     

Induction of enhanced alkaline phosphatase activity in the melano-macrophage centres of Oreochromis aureus (Steindachner) through starvation and vaccination

Employing histochemical methods, alkaline phosphatase activity was studied in the melano-macrophage centres of the spleen of the cichlid fish Oreochromis aureus (Steindachner). Enzyme activity was observed to be very low in normal fish. Prolonged starvation induced an enhanced enzyme response. Starvation followed by antigenic stimulation through an intraperitoneal injection of a bacterial vaccine further elevated the levels of alkaline phosphatase activity. The marked response of the pigment-bearing macrophages to bacterial antigen provides further evidence of the lymphoreticular nature of these pigmented cell aggregates. The association of alkaline phosphatase activity and lipofuscin (the most common pigment in fish melano-macrophage centres) with phagocytic cells has been documented in higher animals including     

Stories about acyl chains

The effect of dietary polyunsaturated fatty acids and alpha-tocopherol supplementation on erythrocyte lipid peroxidation and immunocompetent cells in mice was studied comparatively using seven dietary oils (15% oil/diet, w/w) including fish oil rich in eicosapentaenoic acid (EPA, 20:5, n-3) and docosahexaenoic acid (DHA, 22:6, n-3). A 43% increase in spleen weight, about twice as many spleen cells and no change in the subpopulations of spleen cells, as well as a significant depression of mitogen-induced blastogenesis of both T and B cells in the spleen were observed in mice fed fish oil for 30 days in comparison with soybean oil diet-fed mice. In the fish oil diet-fed mice, membranous lipid hydroperoxide (hydroperoxides of phosphatidylcholine and phosphatidylethanolamine) accumulation as a marker of oxidative senescence in red blood cells (RBC) was 2.7-3.5 times higher than that in mice fed soybean oil, although there was no difference in the plasma phosphatidylcholine hydroperoxide concentration. In spite of the supplementation of alpha-tocopherol to up to 10 times the level in the basal diet, the degeneration of spleen cells and the stimulated oxidative senescence of RBC found by the fish oil feeding could not be prevented. The results suggest that oral intake of excess polyunsaturated fatty acids, i.e. EPA and DHA, in a fish oil diet can lead to acceleration of membrane lipid peroxidation resulting in RBC senescence linked to the lowering of immune response of spleen cells, and that supplementation of alpha-tocopherol as antioxidant does not always effectively prevent such oxidative degeneration as observed in spleen cells and RBC in vivo.     

Evidence for melano-macrophage centres of teleost as evolutionary precursors of germinal centres of higher vertebrates: an immunohistochemical study

The melano-macrophage centres (MMCs) of the haemolymphopoietic organs of teleost fish trap and retain antigens and are closely associated with immunoglobulin-secreting cells. The hypothesis that they are the phylogenetic precursors of the germinal centres of higher vertebrates has been questioned due to their apparent lack of organising cells. In this study the immunoreactivity of MMC cells from spleen and kidney of the teleosts Cyprinus carpio, Odontesthes bonariensis and Solea senegalensis to CNA-42, an antibody usually employed for labelling follicular dendritic cells of higher vertebrates was investigated. Free melano-macrophages and MMCs in the spleens of all three species were labelled by the antibody. This finding adds new evidence to the hypothesis that an evolutionary relationship exists between the MMCs of fish and the germinal centres of many birds and mammals.     

Dietary omega-3 fatty acids enhance the B1 but not the B2 cell immune response in mice with antigen-induced peritonitis

The effects of omega-3 fatty acids on the adaptive immune response have mainly been analysed in vitro with varying results. How omega-3 fatty acids affect the adaptive immune response in vivo is largely unknown. This study examined the effects of dietary fish oil on the adaptive immune response in antigen-induced inflammation in mice, focusing on its effects on B cells and B cell subsets. Mice were fed a control diet with or without 2.8% fish oil, immunized twice with methylated BSA (mBSA) and peritonitis induced by intraperitoneal injection of mBSA. Serum, spleen and peritoneal exudate were collected prior to and at different time points after induction of peritonitis. Serum levels of mBSA-specific antibodies were determined by ELISA and the number of peritoneal and splenic lymphocytes by flow cytometry. The levels of germinal center B cells and IgM⁺, IgG⁺ and CD138⁺ cells in spleen were evaluated by immunoenzyme staining. Mice fed the fish oil diet had more peritoneal B1 cells, more IgM⁺ cells in spleen and higher levels of serum mBSA-specific IgM antibodies compared with that in mice fed the control diet. However, dietary fish oil did not affect the number of peritoneal B2 cells, splenic IgG⁺ or CD138⁺ cells or serum levels of mBSA-specific IgG antibodies in mice with mBSA-induced peritonitis. These results indicate that dietary fish oil can enhance the adaptive immune response, specifically the B1 cell response, which may lead to better protection against secondary infection as well as improvement in reaching homeostasis following antigenic challenge.     

Fish oil diet affects on oxidative senescence of red blood cells linked to degeneration of spleen cells in mice

The effect of dietary polyunsaturated fatty acids and α-tocopherol supplementation on erythrocyte lipid peroxidation and immunocompetent cells in mice was studied comparatively using seven dietary oils (15% oil/diet, w/w) including fish oil rich in eicosapentaenoic acid (EPA, 20:5, n–3) and docosahexaenoic acid (DHA, 22:6, n–3). A 43% increase in spleen weight, about twice as many spleen cells and no change in the subpopulations of spleen cells, as well as a significant depression of mitogen-induced blastogenesis of both T and B cells in the spleen were observed in mice fed fish oil for 30 days in comparison with soybean oil diet-fed mice. In the fish oil diet-fed mice, membranous lipid hydroperoxide (hydroperoxides of phosphatidylcholine and phosphatidylethanolamine) accumulation as a marker of oxidative senescence in red blood cells (RBC) was 2.7–3.5 times higher than that in mice fed soybean oil, although there was no difference in the plasma phosphatidylcholine hydroperoxide concentration. In spite of the supplementation of α-tocopherol to up to 10 times the level in the basal diet, the degeneration of spleen cells and the stimulated oxidative senescence of RBC found by the fish oil feeding could not be prevented. The results suggest that oral intake of excess polyunsaturated fatty acids, i.e. EPA and DHA, in a fish oil diet can lead to acceleration of membrane lipid peroxidation resulting in RBC senescence linked to the lowering of immune response of spleen cells, and that supplementation of α-tocopherol as antioxidant does not always effectively prevent such oxidative degeneration as observed in spleen cells and RBC in vivo.     

家蝇作为饲料添加剂对清远良种鸡肉质和风味的影响

测定蝇蛆和蝇蛹作为饲料添加剂对清远良种鸡肉质和风味的影响,为家蝇Musca domestica L.作为添加饲料改善鸡的肉质和风味,以及代替鱼粉成为新的蛋白质来源提供数据。150只清远鸡雏鸡随机等分成5组,以鱼粉或商业饲料加入不同比例的家蝇幼虫或蝇蛹作为添加饲料加以饲养。结果显示,饲喂不同食料的清远鸡的增重、脾重和法氏囊重影响差异不显著,但饲喂蝇蛆和蝇蛹的鸡群的死亡率明显低于仅饲喂鱼粉或商业饲料的鸡群;饲喂蝇蛆或蝇蛹的鸡肉的系水力(用失水率表示)和风味评价明显提高,氨基酸含量差异显著。说明家蝇幼虫作为动物蛋白可提高鸡的抗疾病能力及改善鸡肉风味。     

Evaluation of immunostimulatory and growth promoting effect of seed fractions of Achyranthes aspera in common carp Cyprinus carpio and identification of active constituents

Immunostimulatory and growth promoting properties of Achyranthes aspera seeds were studied with larvae of common carp Cyprinus carpio. Four experimental diets were prepared using raw (D1) and alcohol (D2), petroleum ether (D3) and 50% aqueous alcohol (D4) extracts of A. aspera seeds. Diet without seed served as control (D5). Fish were fed with test/control diet for 30 days and then immunized with 10 μl of c-RBC. Blood samples were collected 7 days after immunization. Survival (93 ± 3%) of fish was significantly (P < 0.05) higher in D1 diet fed group compared to others. Highest specific growth rate was found in fish fed with diet D2. Significantly (P < 0.05) higher levels of serum protein and albumin were found in D1 and D3 compared to others. Highest serum globulin level was found in D1, which was followed by D3, D2, D4 and D5. Hemagglutination titer level was 5-18 folds higher in diet D3 fed fish compared to others. SGOT and SGPT levels were significantly (P < 0.05) higher in control group compared to the treated groups. Myeloperoxidase activity was significantly (P < 0.05) higher in D1 (2.513 ± 0.27 λ 450 nm) and D3 (2.38 ± 0.07 λ 450 nm) diets fed groups compared to others. The best performance of fish was found in raw A. aspera seeds incorporated diet fed group and the active constituents were identified as ecdysterone and two essential fatty acids linolenic acid and oleic acid.     

Dietary fish oil increases the proportion of a specific neutrophil subpopulation in blood and total neutrophils in peritoneum of mice following endotoxin-induced inflammation

Omega-3 polyunsaturated fatty acids may have beneficial effects in inflammation, where neutrophil migration and activation are of importance. The effects of dietary fish oil on neutrophil numbers and subpopulations in healthy mice and mice with endotoxin-induced inflammation were determined. Mice were fed a control diet with or without 2.8% fish oil, and half of them were injected intraperitoneally with endotoxin. Blood, peritoneal lavage, bone marrow and spleen were collected. Expression of cell surface molecules was analyzed by flow cytometry, and chemokine concentrations were determined by enzyme-linked immunosorbent assay. Dietary fish oil did not alter the proportion of total neutrophils in blood but increased the proportion of a specific subpopulation of neutrophils 48 h following endotoxin administration. This subpopulation of neutrophils expressed higher levels of CD11b, Ly6G and major histocompatibility complex-II, suggesting a different role for these neutrophils in the inflammatory response. Dietary fish oil did not affect neutrophil numbers in the peritoneum of healthy mice, but 12 h after endotoxin administration, there were fewer neutrophils in the peritoneum of mice fed the fish oil diet than in mice fed the control diet. However, 48 h after endotoxin administration, mice fed the fish oil diet had more neutrophils in peritoneum than mice fed the control diet. These results indicate that, although dietary fish oil may delay recruitment of neutrophils from blood to the peritoneum early in inflammation, it has the potential to increase the number of peritoneal neutrophils later, which may be of benefit as impaired neutrophil migration and activation have been associated with immunosuppression late in inflammation.     

Ligula intestinalis (L.) (Cestoda: Pseudophyllidea): plerocercoid-induced changes in the spleen and pronephros of roach, Rutilus rutilus (L.), and gudgeon, Gobio gobio (L.)

The effects of the plerocercoid of Ligula intestinalis (L.) (Cestoda: Pseudophyllidea) on the major lymphoid organs, the spleen and pronephros, of roach, Rutilus rutilus (L.), and gudgeon, Gobio gobio (L.), are described. The spleen of ligulosed roach showed a significant decrease in weight. Differential cell counts suggested this was due to a reduction in erythrocytes, despite significant increases in macrophages and vacuolated granulocytes. The spleen of gudgeon, which consisted almost entirely of erythrocytes, showed a slight reduction in weight in ligulosed fish. In both roach and gudgeon this decrease was independent of parasite burden. Differential cell counts of the pronephros from ligulosed roach revealed a significant decrease in neutrophils and increase in vacuolated granulocytes. In the pronephros of gudgeon, however, cell counts were unaffected by ligulosis. Ultrastructural observations included an apparent disintegration of vacuolated granulocytes and increased pinocytic activity in specialized endothelial cells in the spleens of ligulosed roach. Also, melano-macrophage centres and melano-macrophages increased in the spleen of ligulosed roach and gudgeon, respectively. The marked changes in spleen weight and differential cell counts in ligulosed roach and lack of such changes in ligulosed gudgeon correlate with the differential response to the parasite by these two fish species.     

Synergies between vaccination and dietary arginine and glutamine supplementation improve the immune response of channel catfish against Edwardsiella ictaluri

Channel catfish was used to investigate the enhancement of vaccine efficacy following dietary supplementation with arginine (ARG, 4% of diet), glutamine (GLN, 2% of diet), or a combination of both. After vaccination against Edwardsiella ictaluri, humoral and cellular immune responses, along with lymphoid organ responses were evaluated. E.?ictaluri-specific antibody titers in plasma were higher (P?相似文献   

Daily patterns of mRNA expression of two core circadian regulatory proteins, Clock2 and Per1, and two appetite-regulating peptides, OX and NPY, in goldfish (Carassius auratus)

The aim of this study was to examine the daily hypothalamic mRNA expression profiles for two core circadian regulatory proteins, CLOCK2 and PER1, and for two neuropeptides that regulate wakefulness and food intake, OX and NPY, in goldfish. The profiles were determined for fish at different nutritional states (i.e. fed or unfed on sampling day) and held at different photoperiods (i.e. 16L:8D photoperiod vs. constant light LL). Our results show that under a 16L:8D photoperiod, both fed and unfed goldfish exhibit clear antiphasic daily rhythms of hypothalamic Clock2 and Per1 mRNA expression levels, whereas under LL, daily Clock2 rhythms are seen in both fed and unfed fish while significant rhythms of Per1 mRNA expression only persist in unfed fish. In fish held under 16L:8D, but not under LL, there was significantly higher Per1 expression in fed fish at feeding time than in unfed fish. Daily variations in hypothalamic OX mRNA expression levels with peaks observed prior to both feeding time and the onset of darkness, were displayed under a 16L:8D photoperiod, whereas exposure to LL resulted in lower expression levels with no significant daily variations. Fish held under LL, but not under 16L:8D, showed significant daily variations in NPY mRNA expression with a peak prior to feeding time. Taken together, our results suggest that the mRNA expression of both appetite-regulating and circadian proteins display daily variations and that these patterns can be affected by external cues such as feeding and photoperiod.     

鲤鱼和鲫鱼脾脏显微结构和亚显微结构的研究

鲤鱼和鲫鱼的脾脏中都没有小梁,其红髓和白髓是混合的。脾脏中都有椭圆体,其末端向脾髓开放。脾髓的纤维网眼中缺乏血管壁,椭圆体毛细血管中的血液可直接流入脾髓网状结缔组织的腔隙中。腹腔注射墨汁的鲫鱼,2小时后即可在椭圆体的巨噬细胞中看到碳粒的积累,可见脾脏的过滤作用主要在椭圆体中进行。在注射墨汁的鲫鱼脾脏中,黑色素巨噬细胞中心比未经注射鱼的脾脏明显,且数量增多,它和哺乳动物淋巴结和脾脏中的生发中心结构和功能都不相同。       

饲料中添加丝兰提取物对大菱鲆幼鱼生长和生理及水环境的影响

用丝兰提取物添加量为0.00% (D0)、0.05% (D1)、0.10% (D2)、0.20% (D3)和0.40% (D4)的实验饲料投喂大菱鲆幼鱼(42.200.06) g 60d, 研究其对大菱鲆幼鱼生长性能、体组成、血清免疫代谢指标及养殖水质指标的影响。结果显示, 在饲料中添加丝兰提取物对大菱鲆幼鱼增重率(WGR)、特定生长率(SGR)、饲料系数(FCR)、脏体比(VSI)、肝体比(HSI)、肥满度(CF)均无显著影响(P 0.05)。全鱼、肌肉及肝脏中水分、粗蛋白、粗脂肪含量变化无显著性差异(P 0.05)。D3组幼鱼血清溶菌酶显著高于其他各组(P 0.05), 而D4组各免疫指标明显低于对照组(P 0.05); D3和D4组幼鱼血清的谷丙转氨酶(ALT)、谷草转氨酶(AST)活力显著低于对照组。除A1-6h组外各实验组水体氨氮含量均显著低于对照组(P 0.05), 各实验组亚硝酸盐含量均显著高于对照组(P 0.05), 各实验组总氮、总磷、磷酸盐含量均与对照组无显著性差异(P 0.05)。研究证明, 在饲料中适量添加丝兰提取物能够显著增强大菱鲆幼鱼非特异性免疫能力, 并显著降低养殖水体氨氮含量, 对大菱鲆生长及体组成无显著影响。以非特异性免疫及水体氨氮含量为综合评定指标, 大菱鲆幼鱼饲料中丝兰提取物的适宜添加量为0.20%。       

Dietary fish oil reduces the acute inflammatory response and enhances resolution of antigen-induced peritonitis

Dietary n-3 polyunsaturated fatty acids (PUFA) influence the inductive phase of inflammation but less is known about their effects on the resolution phase. This study examined the effects of dietary fish oil on induction and resolution of antigen-induced inflammation in mice. Mice were fed a control diet with or without 2.8% fish oil, immunized twice with methylated BSA (mBSA) and inflammation induced by intraperitoneal injection of mBSA. Prior to and at different time points after mBSA administration, peritoneal cells were analyzed and expression of surface molecules determined by flow cytometry. Concentration of chemokines, cytokines and soluble cytokine receptors was determined by ELISA. Mice fed the fish oil diet had fewer peritoneal neutrophils, shorter resolution interval and lower levels of pro-inflammatory cytokines and chemokines than mice fed the control diet. In mice fed the fish oil diet there was an early peak in peritoneal levels of the immunosuppressive molecules sIL-6R and TGF-β, that was not seen in mice fed the control diet. In the resolution phase, peritoneal macrophages from mice fed the fish oil diet expressed more of the atypical chemokine receptor D6 and peritoneal TGF-β levels were higher than that in mice fed the control diet. Furthermore, in the late-resolution phase there were more peritoneal eosinophils and macrophages in mice fed the fish oil diet than in mice fed the control diet. These results demonstrate a suppressive effect of n-3 PUFA on the inductive phase of inflammation and indicate an enhancing effect of n-3 PUFA on resolution of inflammation.     

Effects of diets containing soybean meal on trypsin mRNA expression and activity in Atlantic salmon (Salmo salar L)

Atlantic salmon develop subacute enteritis in the distal intestine (DI) when fed diets containing soybean meal (SBM) at high levels, a condition accompanied by increased trypsin activity in the DI intestinal content compared to fish fed conventional fishmeal (FM) based diets. To further investigate the responses of Atlantic salmon to dietary SBM, we measured trypsin activity in intestinal contents, quantified pancreatic trypsin mRNA expression, surveyed trypsin mRNA expression in selected tissues and characterized active forms of trypsin in the intestinal wall and brain. Enzyme measurements showed that trypsin activity in the intestinal content of SBM fed fish was lower in the proximal segments of the intestine, but higher in the DI compared to FM fed fish. The difference in enzyme activity was not reflected in a differential expression of pancreatic trypsin mRNA between fish fed the different diets (FM or SBM). Trypsin mRNA was expressed in 18 different tissues (esophagus, stomach, pancreas, pyloric tissue, midintestine, distal intestine, liver, head kidney, kidney, heart, spleen, thymus, brain, eye, gills, gonads, muscle and skin) but was most prominently expressed in tissues of the gastrointestinal (GI) tract and brain. We report for the first time an upregulation of trypsin-like activity in the DI wall using an in-gel trypsin activity assay, as well as modulated activity in the brain of fish fed SBM. The increased activity in the DI wall may contribute to disease severity and higher trypsin activity in the intestinal content.     

Effect of fasting on nychthemeral concentrations of plasma growth hormone (GH), insulin-like growth factor I (IGF-I), and cortisol in channel catfish (Ictalurus punctatus)

This experiment was conducted to characterize the effect of fasting versus satiety feeding on plasma concentrations of GH, IGF-I, and cortisol over a nychthemeron. Channel catfish fingerlings were acclimated for two weeks under a 12L:12D photoperiod, then fed or fasted for 21 d. On day 21, blood samples were collected every 2 h for 24 h. Weight of fed fish increased an average of 66.2% and fasted fish lost 21.7% of body weight on average. Average nychthemeral concentrations of plasma GH were not significantly different between fed (24.7 ng/mL) and fasted (26.8 ng/mL) fish, but average nychthemeral IGF-I concentrations were higher in fed (23.4 ng/mL) versus fasted (17.8 ng/mL) fish. An increase in plasma IGF-I concentrations was observed in fasted fish 2 h after a peak in plasma GH, but not in fed fish. Average nychthemeral plasma cortisol concentrations were higher in fed (14.5 ng/mL) versus fasted (11.0 ng/mL) fish after 21 d. Significant fluctuations and a postprandial increase in plasma cortisol were observed in fed fish and there was an overall increase in plasma cortisol of both fasted and fed fish during the scotophase. The present experiment indicates little or no effect of 21-d fasting on plasma GH levels but demonstrates fasting-induced suppression of plasma IGF-I and cortisol levels in channel catfish.