Surplus spermine does not protect salmon cells against lipopolysaccharide (LPS)‐induced stress,but excess spermine is eliminated through spermidine/spermine N1‐acetyltransferase (SSAT)

The present trials tested the efficiency of surplus spermine to reduce inflammation and oxidative stress following LPS‐induced stress using an in vitro model of head kidney and liver cells isolated from Atlantic salmon. Spermine did not protect cells from LPS‐induced inflammatory response at either 0.3, 0.6 or 0.9 mM. However, as the gene expression of spermidine/spermine N1‐acetyltransferase (SSAT) increased with increasing spermine concentration, we addressed possible oxidative effects of the increased SSAT using its activator DENSPM or inhibitor of polyamine oxidation of the acetylated polyamines using MDL72527 at a spermine concentration of 0.6 mM. There was no significant effect of DENSPM, but MDL72527 decreased gene expression of GPX‐3 (p = .04), while gene expression of catalase and MnSOD was unaffected by treatment (p = .30 and p = .48, respectively). In conclusion, spermine did not protect cells from LPS‐provoked inflammation. The higher the spermine concentration, the more SSAT producing acetylated spermine occurred. Inhibiting the acetylated polyamine oxidases by MDL72527 improved oxidation status as expected due to a lower endogenous production of H₂O₂ by polyamine and acetylated polyamine oxidases. Probably care should be taken using polyamines or arginine as functional ingredients to avoid any increased oxidation within cells.     

CpG-DNA的功能及应用前景

在细菌及病毒的 DNA中普遍存在的以及人工合成的非甲基化的 Cp G基序能通过 Toll样受体 9能直接活化 B细胞 ,诱导其增殖并抑制其凋亡 ;能促进免疫球蛋白和 IL -6、IL -1 0以及 IL -1 2等细胞因子分泌 ;增加 MHCII类分子和 B7共刺激分子的表达 ;激活单核巨噬细胞及树突状细胞分泌各种 Th1型细胞因子和趋化因子 ;促进淋巴细胞增殖 ,诱发体液和细胞介导的免疫反应。Cp G-DNA还可以间接活化细胞毒性T细胞 ,促进 Ig G2 a及 Ig M的优势表达 ,从而转换免疫反应的类型 (Th 到 Th ) ,因此 Cp G-DNA作为一类有希望的免疫增强剂在增强疫苗免疫效果和抗感染免疫中都有潜在的应用前景。     

Effects of dietary supplementation of a commercial prebiotic Previda® on survival,growth, immune responses and gut microbiota of Pacific white shrimp,Litopenaeus vannamei

A 35‐day feeding trial was conducted to evaluate growth, bacterial populations of the gastrointestinal tract (GIT) and immune responses of Litopenaeus vannamei fed diets containing the commercial prebiotic Previda^®. Diets were formulated to contain Previda^® at 0, 0.2, 0.5, 1.0 or 1.6 g kg^?1 by weight. At the end of the study, differences in weight gain and survival among treatments were not significant (P > 0.05), but denaturing gradient gel electrophoresis analysis revealed that the microbial communities in the GIT changed significantly with the inclusion of dietary Previda^® at different levels. Previda^® was therefore able to selectively modify the microbial communities in the shrimp's GIT. Although individual bacterial species were not identified, enteric populations in shrimp fed the prebiotic at similar levels of inclusion were genetically similar. In addition, shrimp fed Previda^® at 1.6 g kg^?1 responded significantly (P < 0.05) better immunologically with respect to hemocyte phagocytic capacity, haemolymph protein, hyaline cell counts and haemolymph glucose compared with shrimp fed the basal diet. Although shrimp were not exposed to virulent pathogens in this study, the observed upregulation of some of imm‐une responses upon prebiotic supplementation indicates that an improved outcome of such challenges may be anticipated in Previda^®‐fed shrimp under commercial conditions.     

Enzyme treatment of Trypanosoma danilewskyi (Laveran and Mesnil) increases its susceptibility to lysis by the alternative complement pathway of goldfish, Carassius auratus (L.)

This study examined whether in vitro-cultured Trypanosoma danilewskyi were susceptible to lysis in the presence or absence of anti-parasite antibodies and complement. Cultured trypanosomes were resistant to lysis by either immune or non-immune goldfish serum. However, trypanosomes treated with the proteolytic enzyme trypsin, which destroys surface proteins of the parasites, became susceptible to lysis when exposed to either immune or non-immune goldfish serum. The lysis by goldfish serum was dependent on the presence of heat-labile factors and occurred at 4 and 20 degrees C. The lysis was also dependent on the presence of Mg(2+) ions but not Ca(2+) ions. Furthermore, treatment of the parasites with different sialidases did not enhance their susceptibility to lysis by goldfish serum. Trypsinized parasites regained resistance to lysis after at least 6-h cultivation in the absence of trypsin and the restoration of full resistance was observed after 24-h cultivation. The resistance to lysis was abrogated when the protein synthesis inhibitor, puromycin, was added to the cultures. These results suggest that trypsinized trypanosomes were susceptible to lysis by goldfish complement (alternative pathway) and that protective surface proteins of the parasite were required for the resistance of normal trypanosomes to lysis.     

昆虫免疫进化的研究进展

昆虫在长期的进化过程中发展出了一套独特的免疫系统,由细胞免疫和体液免疫组成。随着分子生物学研究技术的发展,人们发现昆虫具有复杂的免疫系统,在病原体识别、信号转导途径及效应机制等方面与哺乳动物均高度相似,这些防御机制有着共同的进化起源。越来越多的研究表明昆虫的免疫具有特异性,不仅有免疫记忆,还能改变自身的免疫细胞去对付不同类型的微生物。     

Effects of dietary inactivated Pediococcus pentosaceus on growth performance,feed utilization and blood characteristics of red sea bream,Pagrus major juvenile

A feeding trial was carried out to determine effects of dietary supplements of inactivated Pediococcus pentosaceus (PP) on growth, feed utilization and blood characteristics of red sea bream. Five diets containing different concentrations [0 (PP0), 1.6 × 10¹⁰ (PPI), 1.6 × 10¹¹ (PPII), 1.6 × 10¹² (PPIII) and 3.2 × 10¹² (PPIV) cells g^?1] of PP were fed to fish for 56 days. At the end of the feeding trial, the obtained results revealed a significant increase (P < 0.05) in weight gain, specific growth rate and skin mucus secretion in all PP‐supplemented groups when compared with control group. Moreover, fish fed PPIII and PPIV diets resulted in significantly higher final body weight, feed efficiency ratio and bactericidal activity than those of fish fed PP0 diet (P < 0.05). Further, fish fed the PPIII diet also had a higher feed intake, protein efficiency retention, haematocrit, mucus lysozyme activity (LA), plasma and serum proteins than those of PP0 fed group (P < 0.05). Similarly, fish fed PPIV diet resulted in a significant increase in serum LA, peroxidase activity and tolerance against low‐salinity stress test (P < 0.05). Considering these promising results, we suggest that the supplementation of PP Prep improves growth and health conditions of Pagrus major.     

Competition of infectious hypodermal and haematopoietic necrosis virus (IHHNV) with white spot syndrome virus (WSSV) for binding to shrimp cellular membrane

Infectious hypodermal and haematopoietic necrosis virus (IHHNV) and white spot syndrome virus (WSSV) are two widespread shrimp viruses. The interference of IHHNV on WSSV was the first reported case of viral interference that involved crustacean viruses and has been subsequently confirmed. However, the mechanisms underlying the induction of WSSV resistance through IHHNV infection are practically unknown. In this study, the interference mechanisms between IHHNV and WSSV were studied using a competitive ELISA. The binding of WSSV and IHHNV to cellular membrane of Litopenaeus vannamei was examined. The results suggested that there existed a mutual competition between IHHNV and WSSV for binding to receptors present on cellular membrane of L. vannamei and that the inhibitory effects of WSSV towards IHHNV were more distinct than those of IHHNV towards WSSV.     

灭活菌苗免疫的南方鲇外周血液细胞免疫指标的变化

以嗜水气单胞菌(Aeromonashydrophila)灭活菌苗为免疫原,以平均体重为86 20g人工饲养的健康南方(Silu rusmeridionalis)为实验对象,通过腹腔注射免疫原(浓度为1 0×108/mL)免疫,剂量为0 2mL/尾,并分别在免疫后的第1、2、4、7、14、21、28天对实验鱼尾静脉采血,进行血液生理指标和血液中吞噬细胞活性的测定。结果显示,嗜水气单胞菌灭活菌苗(FKC)可以增加南方外周血液白细胞数量并引起不同种类白细胞组成比例的变化,提高吞噬细胞的吞噬活性。在免疫后第1、2、4、7、14、21、28天,南方外周血中白细胞数量呈明显上升趋势,其中嗜中性粒细胞、单核细胞、淋巴细胞变化趋势更为明显;而且吞噬细胞活性大为提高,吞噬百分比较对照组平均升高了17 66%,吞噬指数比对照组平均升高了1 16,在免疫后第4天吞噬百分比和吞噬指数均达到最高值,吞噬百分比为48 40%,吞噬指数为4 82;免疫后第21天,吞噬百分比和吞噬指数开始下降,但仍高于对照组。由此表明,使用疫苗后,鱼类非特异细胞免疫在初期(尤其是在第2～7天)明显增强,这是鱼用疫苗在免疫早期保护作用的主要机制之一。     

发酵饲料对中华绒螯蟹幼蟹生长、抗氧化、免疫和蛋白代谢的影响

为研究发酵饲料配伍配合饲料投喂对中华绒螯蟹幼蟹生长性能、抗氧化、免疫机能和代谢的影响,实验分别制备了5组饲料:对照组以幼蟹配合饲料为基础饲料(F0),各实验组在配合饲料基础上额外添加日投喂量5%、10%、15%和20%发酵饲料,并依次命名为F5、F10、F15和F20。150只均重无显著差异的中华绒螯蟹随机分5组,每组设3个重复,每重复10只蟹,分别饲喂对应组饲料,每天饱食投喂1次,养殖周期59 d。在养殖结束后采集血淋巴和肝胰腺以测定抗氧化指标、免疫指标和蛋白代谢酶活性。结果发现,额外投喂发酵饲料后,各组幼蟹的增重率和特定生长率均较对照组显著升高。随着额外投喂发酵饲料量的增加,血淋巴中过氧化氢酶含量先上升后下降,其中F5、F10和F15组较对照组显著升高。各处理组酸性磷酸酶活性高于对照组,但差异不显著。各组总蛋白和球蛋白含量随着发酵饲料额外投喂量提高呈现先升高后降低的趋势,但是各组间差异不显著。此外,额外添加日投喂量20%比例以下的发酵饲料,可以提升肝胰腺中胰蛋白酶、胃蛋白酶、谷草转氨酶、谷丙转氨酶活性。研究表明,额外投喂发酵饲料可以促进中华绒螯蟹幼蟹的生长,还可以提高其抗氧化能力、免疫机能和蛋白代谢能力。