Antimicrobial susceptibility and enrofloxacin resistance of streptococcal bacteria from farmed Nile tilapia,Oreochromis niloticus (Linnaeus 1758) in Thailand

This study examined the antimicrobial susceptibility and mutation(s) in quinolone resistance‐determining regions (QRDRs) in streptococcal pathogens isolated from farmed Nile tilapia Oreochromis niloticus in Thailand. Surveillance of antimicrobial susceptibility in tilapia streptococcal pathogens reveals that Streptococcus agalactiae (n = 97) and Streptococcus iniae (n = 3) from diseased tilapia were susceptible to amoxicillin, florfenicol, sulfamethoxazole/trimethoprim and sulfadimethoxine/ormetoprim, however, only 78 isolates were susceptible to enrofloxacin. Twenty‐two enrofloxacin‐resistant S. agalactiae isolates were further examined for mutations in the QRDRs of gyrA, gyrB, parC and parE genes. Twenty isolates had single base pair changed in the gyrA sequence, C‐242‐T. Point mutations in gyrB, GC‐1135, 1136‐AA and T‐1466‐G, were identified in one isolate. All resistant isolates harboured a mutation in the parC gene, C‐236‐A, while no mutations were observed in the parE gene. The study represented mutations of gyrA and parC genes as marked modification of the enrofloxacin‐resistant S. agalactiae from farmed tilapia. This study is a primary report of the QRDRs mutations associated with fluoroquinolone resistance from streptococcal pathogen in the cultured fish. The phenotypic and genotypic characterization of enrofloxacin resistance S. agalactiae evident in this study has led to an improved regulation of antimicrobial use in Thai aquaculture.     

双酶分步水解制备棉籽多肽工艺条件优化

为高效制备棉籽多肽,采用Alcalase酶和Flavourzyme酶对棉籽蛋白进行双酶分步水解.在前期单因素试验的基础上,通过Plackett-Burrman设计对影响其多肽得率的相关因素进行评估并筛选出具有显著效应的3个因素,然后通过三因素三水平的 Box-Behnken 响应面分析法确定双酶分步水解棉籽蛋白的最优操作条件为:底物浓度(w/v)8.36%,先用Alcalase 酶在温度59.9℃、pH值8.0、酶用量25000 U/g的条件下水解150 min,再用Flavourzyme酶在温度50℃、pH值6.0、酶用量27430 U/g的条件下水解120 min,多肽得率可达到65.07%.     

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.     

鳕碎肉酶解物清除羟自由基作用研究

通过测定鳕碎肉酶解物对Fenton体系产生的羟自由基的清除效果,从海洋蛋白酶YS-894、海洋蛋白酶YS-80、胃蛋白酶和木瓜蛋白酶中,筛选出海洋蛋白酶YS-894作为酶解鳕碎肉制备具有较高清除羟自由基活性酶解物的水解酶;用正交试验L9(34)对该酶的水解条件进行优化,并确定了水解的最佳肉水比。最终工艺条件为40℃、酶解45 m in、pH 9.0、酶解质量分数0.25%、底物∶水=1∶3的条件下进行水解,水解物对羟自由基的清除效果较好,清除率为68.88%。     

Effects of oral administration of colostrum whey in peripartum goat on antimicrobial peptides in postpartum milk

The present study was conducted to examine whether colostrum supplementation in peripartum goats increases the antimicrobial peptides in their milk. Goats were orally administered 2 ml of colostrum whey products (colostrum group) or water (control group) daily, from 2 weeks before until 2 weeks after kidding. Body weights of mothers and kids were measured. Blood, milk, and fecal samples were collected from the mothers, and blood samples were collected from the kids. Concentrations of milk antimicrobial peptides (beta‐defensin, cathelicidin, lactoferrin, S100A7, lactoperoxidase, and immunoglobulin A [IgA]) were determined. IgA and nutritional parameters (glucose, total cholesterol, triglyceride, ketone bodies, and non‐esterified fatty acids) were also determined in the blood of mothers and kids. Milk IgA and lactoferrin concentrations were higher in the colostrum group than in the control group. Conversely, lower milk concentrations of S100A7 were observed in the colostrum group than that in the control group. Plasma IgA concentrations were higher for kids from the colostrum group than for those from the control group. These results suggest that oral administration of colostrum in pregnant goats increases IgA concentration in postpartum milk, which can subsequently improve the health of their kids.     

动物应用抗菌药的风险及防制对策

食品动物因大量应用抗菌药物产生耐药菌株，对动物和人类健康及生态环境造成危害，为此欧盟对多种促生长抗菌药颁布了禁令，导致了动物治疗用抗菌药的急剧增加及畜牧业经济效益的降低。本文就此展开讨论，强调必须开发新型抗菌药物，合理应用抗菌药，加强抗菌药药效学及药动学研究，健全食源性病原菌耐药性监测系统等。     

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。     

酵母多糖对空肠弯曲菌感染獭兔生产性能、载菌量和抗菌肽表达的影响

试验旨在研究酵母多糖对空肠弯曲菌感染獭兔生产性能、载菌量、盲肠和皮肤抗菌肽的影响。选取断奶獭兔180只,随机分为5组,每组6个重复,每个重复6只。对照组饲喂基础日粮,其他4组空肠弯曲菌感染,并于基础日粮中分别添加0、200、400、600 mg/kg酵母多糖。饲养试验起始日诱导空肠弯曲菌感染,饲养试验持续35 d。结果显示,空肠弯曲菌感染降低了獭兔结束体重、日增重和日采食量,提高了料重比（P < 0.05）,添加酵母多糖改善了这些指标（P < 0.05）,且日增重和料重比达到了对照组水平。酵母多糖降低了獭兔盲肠食糜、皮肤、肝脏、脾脏的空肠弯曲菌载菌量（P < 0.05）。空肠弯曲菌感染诱导内源性抗菌肽的Cathelicidin、Galectin-3和LEAP2基因表达量显著上调（P < 0.05）,对DEFA4基因无显著影响,而添加酵母多糖对基因表达量具有进一步上调作用（P < 0.05）。酵母多糖添加剂量与结束体重、日增重呈现线性和平方效应关系（P<0.05）;与皮肤、肝脏和脾脏载菌量呈现线性效应（P<0.05）;与盲肠和皮肤抗菌肽Cathelicidin和Galectin-3基因表达水平呈现线性效应（P<0.05）;与盲肠抗菌肽Galectin-3、LEAP2和DEFA4基因呈现平方效应（P<0.05）。表明日粮添加酵母多糖可以降低空肠弯曲菌感染獭兔组织载菌量,提高内源抗菌肽活性,不影响獭兔生产性能。     

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.     

大豆小分子肽增强小鼠免疫力的实验研究

为通过动物实验检测大豆小分子肽(分子量300～700 Da)对免疫系统的影响及其增强免疫力的作用,分别给受试组小鼠经口灌胃低、中、高剂量(分别为3、6、9 g.kg-1.d-1)的大豆小分子肽溶液30 d后,进行各项免疫指标的检测。结果显示:大豆小分子肽能促进小鼠脾淋巴细胞的转化和迟发型变态反应,提高抗体生成细胞数和血清溶血素水平,增强腹腔巨噬细胞的吞噬能力;但对小鼠的NK细胞活性,单核—巨噬细胞的碳廓清能力,体重及免疫器官重量均无显著影响。这表明,大豆小分子肽具有增强机体免疫力的作用。