Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.     

肽在瘤胃内的代谢和利用

微生物蛋白降解作用形成的多肽经过一系列肽酶最终转化为氨基酸,其速度取决于多肽的化学组成和N端结构.大多数情况下,瘤胃肽在栖瘤胃拟杆菌分泌的二肽基肽酶作用下,从多肽N未端释放二肽而不是游离氨基酸.通过抑制栖瘤胃拟杆菌的二肽基肽酶活性或该菌对肽吸收率可以减慢混合瘤胃群落对肽的水解速率.肽和氨基酸刺激瘤胃微生物的生长,并且对许多种菌的最佳生长是必需的.纤维分解菌是个例外.当然,细菌蛋白质合成时利用氨与肽和氨基酸的比例也取决于它们的浓度.     

抗菌肽与抗生素的体外抗菌效果比较

应用由 Hancock实验室改进的微量肉汤稀释法测定了 cecropin P1、cecropin A、magainin 2、defensin 1、bactenecin、lactoferricin和 indolicidin等 7种抗菌肽和盐酸金霉素、去甲万古霉素、土霉素、强力霉素等 4种抗生素的体外抗菌活性 ,并且使用薄层琼脂糖孔穴扩散法比较了抗菌肽中 cecropin P1和 cecropin A与土霉素和呋喃唑酮对大肠杆菌 K88的抑菌效果。结果发现 ,抗菌肽与抗生素一样对几种革兰氏阳性菌和阴性菌都有不同程度的抗菌效果。其中 cecropin A、cecropin P1和 defensin1对大肠杆菌的 2个菌株 ATCC2 5 92 2和 K88的抗菌活性高于抗生素 ,defensin1是各种抗菌肽中对金黄色葡萄球菌抗菌效果最好的。抗生素对试验用金黄色葡萄球菌的抑制活性总体上要好于抗菌肽。与抗生素相比 ,cecropin A、cecropin P1和 indolicidin对猪霍乱沙门氏菌和鼠伤寒沙门氏菌具有更好的抗菌效果。另外 ,抗菌肽的抑菌圈边缘十分清晰 ,而抗生素的整个抑菌圈都比较模糊且界线不明。因此 ,从抗菌效果方面考虑 ,抗菌肽可以代替抗生素用于疾病的预防和治疗     

A review of antimicrobial peptides: defensins and related cationic peptides

Cationic antimicrobial peptides are present throughout the plant and animal kingdoms and bear striking structural and functional similarities across species lines. They provide primitive, nonspecific means of combating a variety of bacteria, fungi, enveloped viruses, and protozoa. Some are also cytotoxic against host cells, including neoplastic cells. Cationic antimicrobial peptides may play various roles in inflammation and tissue repair. Antimicrobial peptides are found in epithelial tissues regularly exposed to microbial attack as well as in cells whose primary function is defense against potential pathogens. They constitute an important part of the nonoxidative antimicrobial arsenal of leukocytes. They are preformed and/or readily synthesized when the cells are stimulated by exposure to pathogens. They exert their effects directly by inserting into membranes of target cells and forming ion channels which increase membrane permeability; however, antimicrobial peptides can also act as opsonins to facilitate phagocytosis. Resistance to defensins is a virulence factor for organisms such as Salmonella sp. The study of cationic antimicrobial peptides is increasing our understanding of innate immunity, inflammation, and the pathogenesis of genetic diseases such as specific granule disease in humans. Therapeutic applications of antimicrobial peptides are currently under investigation.     

昆虫抗菌肽生物学活性及其应用前景

昆虫抗菌肽是昆虫免疫后存在于血淋巴中的一类活性肽 ,具有广谱的抗菌、抗病毒、抑制肿瘤的生物活性 ,具有很高的应用潜力。本文主要介绍昆虫抗菌肽的类型、生理活性、基因的克隆与表达及在动物科学中的应用前景     

世界动物卫生组织抗生素耐药性控制策略研究

世界动物卫生组织(OIE)是动物卫生领域的权威国际组织。通过研究OIE相关标准和策略,为我国进一步加强兽用抗生素管理,更好地防治抗生素耐药性风险提供参考意见。     

Effects of oral orbifloxacin on fecal coliforms in healthy cats: a pilot study

The study objective was to determine the effect of oral orbifloxacin (ORB) on antimicrobial susceptibility and composition of fecal coliforms in cats. Nine cats were randomized to two groups administered a daily oral dose of 2.5 and 5.0 mg ORB/kg for 7 days and a control group (three cats per group). Coliforms were isolated from stool samples and were tested for susceptibilities to ORB and 5 other drugs. ORB concentration in feces was measured using high-performance liquid chromatography (HPLC). The coliforms were undetectable after 2 days of ORB administration, and their number increased in most cats after termination of the administration. Furthermore, only isolates of Escherichia coli were detected in all cats before administration, and those of Citrobacter freundii were detected after termination of the administration. E. coli isolates exhibited high ORB susceptibility [Minimum inhibitory concentration (MIC), ≤0.125 µg/ml] or relatively low susceptibility (MIC, 1−2 µg/ml) with a single gyrA mutation. C. freundii isolates largely exhibited intermediate ORB susceptibility (MIC, 4 µg/ml), in addition to resistance to ampicillin and cefazolin, and harbored qnrB, but not a gyrA mutation. HPLC revealed that the peaks of mean concentration were 61.3 and 141.0 µg/g in groups receiving 2.5 and 5.0 mg/kg, respectively. Our findings suggest that oral ORB may alter the total counts and composition of fecal coliform, but is unlikely to yield highly fluoroquinolone-resistant mutants of E. coli and C. freundii in cats, possibly because of the high drug concentration in feces.     

Outpacing the resistance tsunami: Antimicrobial stewardship in equine medicine,an overview

Antimicrobial stewardship (AMS) is the term increasingly used to describe the multiple approaches needed to sustain the efficacy of antimicrobial drugs in the face of the increasing development and spread of antimicrobial resistance in bacterial pathogens, and the global crisis in medicine that it is engendering. The concept and the practices associated with AMS continue to evolve but the general approach is a dynamic and multifaceted one of continuous improvement based on reducing, improving, monitoring and evaluating the use of antimicrobials so as to preserve their future efficacy and to protect human and animal health. Using many equine examples, this basic overview discusses the multiple and interacting elements of AMS: Practice guidelines, infection control and prevention, clinical microbiology, resistance and use surveillance, dosage, pharmacokinetics and pharmacodynamics, regulation, education and owner compliance, leadership, coordination and measurement. There have been impressive advances in recent years in reporting and analysis of AMR in horses, in the scrutiny and assessment of how antimicrobial drugs are used in horses and in identification of areas for improvement including dosing, surgical prophylaxis, infection control, development of practice standards and the use of clinical microbiology. Antimicrobial stewardship is taking shape as we start to see the emergence of evidence-based recommendations but far more is required. Containing and even rolling back AMR will need the continued engagement of practitioners, equine national and international practitioner organisations, researchers and educators in the academic community, horse owners, regulators and others.     

油菜籽来源抗氧化物质研究进展

油菜籽中含有多种具有抗氧化活性的物质，为抵御机体的氧化损伤起到了积极作用。本文综述了油菜籽来源的诸多抗氧化活性成分及抗氧化活性的评价方法。油菜籽主要的抗氧化成分有肽类、多酚类、多糖类、生育酚和其它抗氧化成分，其中的肽类主要有三种产生方式：非消化酶酶解、消化酶酶解和发酵。针对油菜籽来源的活性物质抗氧化活性的评价方法，包括体外测定方法（化学检测）和细胞内抗氧化实验和动物实验。综述还探讨了油菜籽抗氧化活性成分研究中存在的问题和解决方法，旨在为油菜籽活性成分的开发和利用提供参考。