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.     

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.     

兽用抗寄生虫药物新剂型及其新技术的研究进展

兽用抗寄生虫药物由于使用频繁,其给药技术研究一直备受重视.目前,兽用抗寄生虫药物新剂型和新技术的研究主要集中在缓释控释制剂和脂质体制剂,其次是透皮给药系统、微囊、微球、环糊精包合物、固体分散体等.但这些新剂型和新技术大多还处于研究阶段,在处方设计或制剂工艺方面都还存在一些问题,今后应加强这些新剂型的工艺和技术研究,以真正发挥这些新剂型的优点.     

鸡抗球虫药的体外筛选综述

抗鸡球虫药的筛选分为体内筛选和体外筛选。体外筛选具有简易、迅速、准确和经济的优点。本文在前人大量工作的基础上，总结了体外筛选的全过程，并分述了细胞培养筛选法和鸡胚培养筛选法。     

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.     

规模化蛋鸡场病原菌溯源与生物安全防控研究

细菌病严重威胁蛋鸡健康,以抗生素为主导的细菌病传统防控方式会导致病原菌耐药性严重、鸡蛋药物残留,影响蛋品安全。要保证蛋鸡产蛋期不用抗菌药物而又能生产出"无菌、无抗"的鸡蛋,是国内外技术难题。本团队以规模化蛋鸡场全封闭鸡舍为研究对象,基于PFGE、MLST、全基因组测序等技术,探明了蛋鸡病原菌的种类及基因组特征;对病原菌的耐药性研究表明,蛋鸡病原菌多重耐药严重,依靠药物防控的方式亟待转变;对规模化蛋鸡场细菌溯源研究结果表明,鸡舍生物媒介(鼠、苍蝇等)和非生物媒介(空气、饮水、饲料、鸡粪等)是蛋鸡病原菌的重要传染来源和传播途径;对规模化蛋鸡场生物安全量化评价结果表明,外部和内部生物安全权重高达92.2%。根据溯源和生物安全权重分析结果,提出了在规模化全封闭蛋鸡舍将蛋鸡细菌病从药物防控转变为生物安全防控的新策略;创新了规模化蛋鸡场细菌病防控的系统技术,实现蛋鸡规模养殖整个产蛋期不用抗生素,为生产"无菌、无抗"安全鸡蛋提供技术支撑。     

肉牛源细极链格孢菌的分离鉴定

为鉴定肉牛皮肤病病原及其致病性并筛选其敏感药物,本研究取患皮肤病肉牛病变部位皮屑、毛发和痂皮进行病原菌的分离,并对分离菌株进行了形态学鉴定、ITS序列分析、小鼠致病性试验和药敏试验。结果显示分离到1株形态特征与细极链格孢菌极其相似的菌株(PY2-1-2);经PCR扩增、测序和BLAST序列比对,结果显示分离株(PY2-1-2)的ITS基因序列与细极链格孢菌(MG975630.1)的ITS基因序列的相似性为99%,分离株PY2-1-2的ITS基因序列长度为543 bp,该片段包括ITS1、5.8S rDNA和ITS2的全部基因序列以及18S rDNA和28S rDNA的部分基因序列,提交GenBank(MH656780.1)。根据形态学结合ITS基因序列分析结果确定该分离株为细极链格孢菌。小鼠致病性试验结果显示该菌对小鼠有致病性。药敏试验结果显示:该菌对灰黄霉素、氟康唑、伊曲康唑、酮康唑的最小抑菌浓度(MIC)值分别为0.5μg/mL、4μg/mL、1μg/mL、1μg/mL。本研究为今后进一步探究细极链格孢菌引起的皮肤病诊治提供科学有效的参考。