生物活性肽调节断奶仔猪肠道健康

断奶是养猪业中最具挑战性的阶段,对仔猪生长性能及之后的发育有着重要的影响。断奶对仔猪的肠道发育和健康带来了极大的挑战,可能会导致采食量减少、生长速率降低、发病率和死亡率增加。采取有效的营养策略可改善仔猪肠道健康和最大限度地提高断奶仔猪的产量。本文综述了生物活性肽通过仔猪肠道免疫、肠道微生物稳态及肠-脑轴的调节仔猪肠道健康,为加深生物活性肽的开发和利用提供参考。     

Structure and function of antimicrobial peptide penaeidin-5 from the black tiger shrimp Penaeus monodon

The gene for penaeidin-5, an antimicrobial peptide comprising 55 amino acids, was isolated from the hemocyte of black tiger shrimp (Penaeus monodon). RT-PCR expression tests revealed that penaeidin-5 was produced in hemocytes, gills, the intestine and muscle. Western blot analysis confirmed the panaeidin-5 was aboundantin hemocytes, the intestine and hemolymph. Immunohistochemistry revealedpenaeidin-5 in the cuticle and gills that are considered primary defense barriers. The deduced amino acid sequence of penaeidin-5 included a proline-rich N-terminal domain and a carboxyl-domain that contained six cysteine residues. Circular dichrosim analysis revealed an -helix in its secondary structure and the predicted 3D structure indicated two-disulfide bridges in the -helix. Based on the sequence of penaeidin-5 peptide cDNA, synthetic penaeidin-5 was prepared to carry out functional tests. The synthetic peptide had efficient bacteriostatic and bactericidal activity against Aerococcus viridans, and also inhibited the growth of two filamentous fungi, Fusarium pisi and Fusarium oxysporum. To measure penaeidin-5 in vivo, black tiger shrimp were challenged with Vibrio alginolyticus and A. viridans. At 3 h post-challenge, penaeidin-5 was induced and bacterial numbers decreased significantly by 12 h and 24 h.     

小肽制品对断奶仔猪生长性能和免疫机能的影响及机理研究

本研究以杜大长早期断奶仔猪为对象,分2次试验探讨了小肽制品对早期断奶仔猪的添加效果。试验1设计4个小肽制品的添加水平(0‰、2‰、4‰、6‰),探讨其对仔猪生长性能的影响,以此选择最佳添加量。试验2以试验1选出的最佳水平为添加量,研究小肽制品对仔猪生长性能和部分血液生理生化指标的影响,进一步探讨小肽作用的某些机理。试验1结果:添加2‰小肽制品的试验仔猪与对照组相比,日采食量和日增重分别提高了15.5%和17.3%(P<0.05);而随着添加量的增加,试验仔猪的生长性能呈下降趋势。故确定小肽制品的最佳添加量为2‰。试验2结果:以添加2‰小肽制品的日粮饲喂仔猪,试验仔猪的日采食量和日增重分别增加了15.7%(P<0.01)和18.1%(P<0.01),料重比和腹泻率分别减少了2.1%(P<0.05)和54.8%(P<0.05),这与试验1结果基本一致;血清总蛋白浓度和球蛋白浓度分别增加了45.8%(P<0.01)和19.4%(P<0.05),尿素氮浓度减少了39.6%(P<0.05),白球比降低了33.9%(P<0.05),谷草转氨酶、谷丙转氨酶、碱性磷酸酶分别增加了28.2%(P<0.05)、26.6%(P<0.05)和39.9%(P<0.01),乳酸脱氢酶活性降低了22.1%(P<0.05)。以上结果显示,小肽制品能显著提高蛋白质的沉积率,提高断奶仔猪的生长性能,增强其免疫机能。     

团头鲂血清抗病毒半乳糖凝集素样蛋白的分离与鉴定

将草鱼出血病病毒(GCHV)的结构蛋白与Sepherose-4B偶联制备亲和柱，经亲和层析从团头鲂血清中分离到一种蛋白，能凝集兔红细胞，其血凝性依赖于β巯基乙醇的存在。红细胞凝集抑制实验证实该蛋白与D半乳糖有最高的亲和力，其次为甘露糖。梯度PAGE测得其分子量约为240000，SDS梯度PAGE证实该蛋白由分子量为15000的单个亚基组成，亚基之间没有共价连接。以上特性与半乳糖凝集素相符，故称其为团头鲂抗病毒半乳糖凝集素样蛋白。抗体阻断试验证实，此种凝集素样蛋白是团头鲂血清抗病毒及红细胞凝集活性的主要成分。N末段的氨基酸序列为Lys-Val-Asn-Leu-Asp-Glu-Lys-Cys-Pro-Phe，检索GenBank，未发现与这一段序列同源的蛋白质。     

Influence of enteric helminths on the distribution of intestinal endocrine cells belonging to the diffuse endocrine system in brown trout,Salmo trutta L

The presence of intestinal helminths in the alimentary canal of brown trout, Salmo trutta L., can alter the number of cells that synthesize modulatory peptides. A total of 167 brown trout were collected from tributaries of the River Brenta (northern Italy), of which 119 (71.3%) specimens were infected with enteric helminths, 28 with the acanthocephalan Pomphorhynchus laevis Müller, 1776 with intensity of infection ranging from 1 to 162 (18.57 +/- 30.79) worms per host and 67 fish with the cestode Cyathocephalus truncatus Pallas, 1781. Intensity of infection with C. truncatus ranged from 1 to 85 (6.87 +/- 12.59) per fish. In 24 fish there were concurrent infections of both species of helminths. The caecal and middle regions of the intestine were the most heavily parasitized. Immunohistochemical tests showed a decrease in endocrine cells (ECs) of the diffuse endocrine system (DES) positive to gastrin, cholecystokinin-8, bombesin and secretin antisera in the intestine of the infected trout. The number of ECs immunoreactive to anti-glucagon serum did not show differences in the digestive tract of uninfected brown trout and in conspecifics parasitized with P. laevis. The density of cells containing glucagon-like material was low in the fish parasitized with C. truncatus. The results suggest that endoparasitic helminths induce alterations in the DES of infected S. trutta.     

日本鳗鲡脾脏抗菌肽抗菌活性检测方法的比较

本试验旨在建立一种在日本鳗鲡脾脏抗菌肽分离纯化过程中简便、灵敏的检测其抗菌活性的方法。试验以日本鳗鲡脾脏分子质量小于10 ku的蛋白质为抗菌肽样品,选择琼脂板孔穴扩散法和微孔液体培养法2种传统方法及微量液体培养法为检测方法,比较这3种方法在抗菌肽对3种鳗鲡常见致病菌株(迟钝爱德华菌、气单胞菌、嗜水气单胞菌)抗菌活性检测中的灵敏度和优缺点。结果表明:与琼脂板孔穴扩散法及微孔液体培养法相比,微量液体培养法简便、易于观察结果,具有最高的灵敏度且样品用量最少,适合在鳗鲡脾脏抗菌肽分离纯化过程中跟踪检测抗菌活性,尤其适合分离纯化后期获得微量单一样品抗菌活性的检测。由此得出,微量液体培养法适用于检测日本鳗鲡脾脏抗菌肽的抗菌活性。     

抗菌肽的作用机理及其在养殖中的应用进展

抗菌肽是生物体内诱导产生的一类具有抗菌作用的生物活性肽,具有广谱的抗菌活性以及其他重要的生物学功能,被用作饲料添加剂广泛应用于畜禽生产中。文章综述了抗菌肽的理化特性、作用机理、生物学功能及其在养殖中的应用现状。     

凡纳滨对虾在微囊藻毒素(MC-LR)诱导下3种抗菌肽基因的表达量分析

蓝藻水华可产生大量藻毒素,其中以微囊藻毒素危害最大。研究表明微囊藻毒素可影响包括虾类在内的多种水生生物的生长和繁殖,亦可导致虾类免疫系统受损。本研究通过实时荧光定量PCR(qRT-PCR)技术检测了在微囊藻毒素MC-LR作用下,凡纳滨对虾(Litopenaeus vannamei)体液免疫中3种主要的抗菌肽基因(Penaeidin 3、Crustin和ALF)的表达量变化情况,发现Penaeidin 3和ALF受到MC-LR影响较大,Crustin基因表达量虽有所上调,但变化不大。同时,通过再次注射鳗弧菌(Vibrio anguillarum)对3种抗菌肽在MC-LR影响下抗菌能力的检测,发现3种抗菌肽表现出相对一致的变化,即表达量均在前期下调,4 h后逐渐上调,在MC-LR处理组中的表达量低于对照组的趋势。结果表明MC-LR可能通过直接或间接的方式影响对虾3种抗菌肽基因的表达,对免疫系统造成损伤,致使对虾抗病能力减弱;这一结果表明,长期发生铜绿微囊藻水华的虾池对虾存活率下降,可能与微囊藻毒素影响对虾抗菌肽基因的表达有关。     

Vasonatrin peptide attenuates injuries of dopaminergic neurons induced by 1-methyl-4-phenylpyridinium

AIM：To investigate the neuroprotective effects of vasonatrin peptide (VNP) on the injury of dopaminergic neurons induced by 1-methyl-4-phenylpyridinium (MPP+). METHODS：Cultured dopaminergic neurons from the mouse ventral mesencephalon were exposed to MPP+, and the effects of VNP on the neurotoxicity of MPP+ were eva-luated by cell viability analysis and immunofluorescence staining. Various kinds of agonists and antagonists were used to clarify the mechanism underlying the effects of VNP. RESULTS：MPP+ caused injuries in the dopaminergic neurons. VNP significantly increased the viability, axon number and axon length of the dopaminergic neurons. The MPP+-induced depolymerization of β-tubulin Ⅲ was also attenuated by the treatment with VNP. In addition, VNP significantly increased the intracellular levels of cGMP. These effects of VNP were mimicked by 8-Br-cGMP (a cell-permeable analog of cGMP), whereas inhibited by HS-142-1 ［the antagonist of the particulate guanylyl cyclase-coupled natriuretic peptide receptors (NPR)］, or KT-5823 ［a cGMP-dependent protein kinase (PKG) inhibitor］. CONCLUSION：VNP attenuates the neurotoxicity of MPP+ via guanylyl cyclase-coupled NPR/cGMP/PKG pathway, indicating that VNP might be a new effective reagent in the treatment of neural degeneration of dopaminergic neurons in Parkinson disease.     

抗菌肽基因异源表达系统研究进展

抗菌肽是机体天然免疫系统的重要组成部分,因具有抗菌谱广、活性稳定、不易产生耐药性等优点而在食品、饲料、农业、医药领域具有重要的应用价值.由于化学合成的成本较高,利用基因工程技术构建高效的生物表达系统成为一个新的途径.综述了抗菌肽在细菌、酵母、植物、动物等表达系统中异源表达的研究进展,为抗菌肽的研究应用提供理论依据.