Identifying and Characterizing a Novel Peritrophic Matrix Protein (MdPM-17) Associated With Antibacterial Response From the Housefly,Musca domestica (Diptera: Muscidae)

Peritrophic matrix/membrane (PM) critically prevents the midgut of insects from external invasion by microbes. The proteins in the peritrophic membrane are its major structural components. Additionally, they determine the formation and function of this membrane. However, the role of PM proteins in immune regulation is unclear. Herein, we isolated a novel PM protein (MdPM-17) from Musca domestica larvae. Further, the function of MdPM-17 in regulating host innate immunity was identified. Results showed that the cDNA of MdPM-17 full is 635 bp in length. Moreover, it consists of a 477-bp open reading frame encoding 158 amino acid residues. These amino acid residues are composed of two Chitin-binding type-2 domain (ChtBD2) and 19 amino acids as a signal peptide. Moreover, tissue distribution analysis indicates that MdPM-17 was enriched expressed in midgut, and moderate levels in the fat body, foregut, and malpighian tubule. Notably, MdPM-17 recombinant protein showed high chitin-binding capacity, thus belongs to the Class III PM protein group. MdPM-17 protein silencing via RNA interference resulted in the expression of antimicrobial peptide (defensin, cecropins, and diptericin) genes, and this occurred after oral inoculation with exogenous microbes Escherichia coli (Enterobacteriales:Enterobacteriaceae), Staphylococcus aureus (Bacillales:Staphylococcaceae), and Candida albicans (Endomycetales:Saccharomycetaceae)). Therefore, all the antimicrobial peptide (AMP) gene expression levels are high in MdPM-17-depleted larvae during microbial infection compared to controls. Consequently, these findings indicate that MdPM-17 protein is associated with the antibacterial response from the housefly.     

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。     

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.     

Effects of antimicrobial peptides on growth,morphology of foregut villi and related genes mRNA expression in the common carp (Cyprinus carpio)

A 60‐day feeding trial was conducted to examine the effects of different levels (0, 100, 200, 400 and 600 mg/kg) of antimicrobial peptides on growth, protease activity of foregut, the morphology of foregut villi and related genes mRNA expression level in the common carp (Cyprinus carpio). The results showed that the feed of antimicrobial peptides promote common carp growth, and the optimal dosage of antimicrobial peptides is 200–333 mg/kg in the common carp feed. The protease activity of 200 and 400 mg/kg groups were significantly higher than the control and other groups (p < 0.05). The foregut villus height with 100, 200 and 400 mg/kg antimicrobial peptide groups were significantly higher than control group (p < 0.05). The crypt depth of 200 and 400 mg/kg antimicrobial peptide groups were significantly lower than control group (p < 0.05). The ratio of villus height and crypt depth of 100, 200 and 400 mg/kg antimicrobial peptide groups were significantly higher than control group (p < 0.05). The ratio with 600 mg/kg group was significantly lower than the control group (p < 0.05). The IGF‐I gene expression level of 200 mg/kg and 400 mg/kg groups were significantly higher than the control group and 600 mg/kg group (p < 0.05). The IL‐1β gene expression level of 100 mg/kg and 200 mg/kg groups were significantly higher than the control group (p < 0.05). These results indicated up‐regulation of growth and immune related genes in antimicrobial peptides fed common carp. Correlation analysis showed that IGF‐I mRNA and IL‐1β mRNA were positively correlated with SGR. IL‐1β mRNA and FCR were significantly negative correlated. It indicated that growth and immune gene common regulated the growth of the carp under antimicrobial peptides intervention. In conclusion, antimicrobial peptides can improve growth and related genes mRNA expression in the common carp. Further studies using molecular biological technique or immunologic methods are required to conclude that antimicrobial peptides are beneficial in common carp.     

Effects of antimicrobial peptides on growth,feed utilization,serum biochemical indices and disease resistance of juvenile shrimp,Litopenaeus vannamei

This experiment was conducted to assess the effects of antimicrobial peptides at different levels (Diet 1 (0%), Diet 2 (0.1%), Diet 3 (0.2%), Diet 4 (0.4%), Diet 5 (0.6%) and Diet 6 (1%)), on growth, serum biochemical indices and antioxidant effect, feed utilization and disease resistance in Litopeneaus vannamei. There were four replicates in each group in the experiment (mean weight = 0.21 ± 0.00 g) and also fed with their respective diets for 8 weeks. Growth performance compared with the control group (0%) significantly increased at first and then decreased among treatment groups (p < .05) whereas the survival rate ranged from 78% to 96%. No significant differences were observed in terms of moisture, crude protein and ash content, but there was a significant increase in crude lipid (p < .05). In serum, acid phosphatase, alkaline phosphatase, polyphenol oxidase, triglyceride, glucose and total cholesterol changed as compared with 0%. Total antioxidant capacity, glutathione peroxidase, catalase, superoxide dismutase and malondialdehyde were also different from 0%. Disease resistance was increased in shrimp among treatment groups with 0.4% recording the lowest mortality percentage of 37% after the challenge test. The results from the present study suggested that supplementation of AMP at 0.4% in shrimp diet can improve growth performance, antioxidants activities and innate immune response of Pacific whiteleg shrimp.     

一种安全有效提高虹鳟苗种成活率的技术

为安全有效地提高虹鳟苗种成活率,应用1种生物制剂——抗菌肽对虹鳟苗种进行浸泡处理。通过比较苗种成活率,检测SOD、ALP、CAT、IL、IFN、TNF等6种免疫相关酶表达量,采用扫描电镜检测苗种体表细菌情况,确定抗菌肽浸泡对虹鳟成活率的影响。试验结果显示,自破膜期开始,用40%抗菌肽浸泡10次,试验组6种免疫相关酶表达量明显高于对照组,试验组苗种的成活率高达95%以上,对照组苗种的成活率则低于30%。电镜观察结果显示,试验组仔鱼体表细菌数量显著减少。试验结果表明,用抗菌肽浸泡法能显著提高虹鳟苗种成活率。     

养殖大菱鲆肠道中大菱鲆弧菌的分离鉴定及药敏试验

2014年11月,大连地区某大菱鲆养殖场发生病害并伴有死亡,主要症状为吻部下颌明显出血,腹腔积水,肝脏充血,肠道出血有白便。从患病大菱鲆肠道中分离出1株可在硫代硫酸盐—柠檬酸盐—胆盐—蔗糖琼脂培养基上生长的菌株HZ-C1,人工回接感染试验证明其对健康大菱鲆具有较强致病性。通过形态学观察、16SrDNA序列同源性分析及生理生化试验确定菌株HZ-C1为大菱鲆弧菌。药敏检测结果显示,大菱鲆弧菌HZ-C1对氯霉素、丁胺卡那及头孢曲松等11种抗生素敏感,但对青霉素、四环素及诺氟沙星等15种抗生素耐药。本研究为大连地区养殖大菱鲆细菌性病害防治提供了一定病原学依据。     

Assessment of the Antibacterial Effect of Chitosan Coated with Heracleum persicum Oil on Rainbow Trout Quality

Today, due to the rapid spoilage of fish, the use of natural preservatives is a priority over those of synthetic varieties. Also, the natural antimicrobial effects of essential oils can help to increase shelf life. There are very few studies concerning the use of essential oils in this regard. In the present study, the antibacterial effect of chitosan coated with Heracleum persicum oil was investigated on the quality of rainbow trout. The control and the coated fish samples were analyzed periodically by generalized estimating equation (GEE) for total viable count (TVC), psychrophilic bacteria, total volatile base nitrogen (TVB-N), polyunsaturated fatty acids (PUFA), texture, taste, odor, and overall acceptance parameters. The results showed that by increasing the amount of chitosan, TVC, psychrophilic bacteria, and TVB-N decreased while PUFA, texture, taste, odor, and overall acceptance parameters increased.     

牛角瓜根的抑菌活性研究

以牛角瓜根为实验材料,采用索氏提取法,以4种不同溶剂(石油醚、氯仿、乙酸乙酯、甲醇)对牛角瓜的根进行提取,并采用纸片琼脂扩散法对粗提物的抑菌活性进行了评价,供试菌包括白色念珠菌等6种真菌和金黄色葡萄球菌等7种细菌。结果表明:石油醚粗提物对枯草芽孢杆菌、白色念珠菌、黑曲霉菌、大肠埃希氏菌都表现出良好的抑菌活性,且抑菌效果接近或优于阳性对照氟康唑,对尖孢镰刀菌和鲍曼不动杆菌也表现出良好的抑菌活性;氯仿粗提物对白色念珠菌及粪链球菌表现出良好的抑菌活性;乙酸乙酯粗提物对粪链球菌表现出最好的抑菌活性;甲醇粗提物的抑菌活性较弱。     

Antimicrobial Susceptibility and Characterization of Outer Membrane Proteins of Aeromonas hydrophila Isolated in China

Aeromonas hydrophila isolates from clinical cases （n=43） were tested against 8 antimicrobial agents and typed by outer membrane protein （OMP） pattern by using sodium dodecyl sulfate gel electrophoresis. All isolates were resistant to ampicillin （MICs, ≥16 μg mL-1） and sulfamonomethoxine （MICs≥64 μg mLl）, but susceptible to norfloxacin （MICs,≤0.5 μg mL-1）. There was a high incidence of resistance to erythromycin （90.70%） and tylosin （93.02%）, while a low incidences of resistance to ciprofloxacin （2.33%）, enrofloxacin （2.33%） and florfenicol （4.65%）. Six different outer membrane protein patterns were found among 34 isolates by analyzing proteins in the range of 22 to 50 kDa, other than 9 isolates with their respective profiles. The strains with the similar OMP profiles had similar resistances. Compared with the other strains from the same OMP patterns, NB-1, A.Pun and MR-1 had lacked the proteins in the range of 30 to 45 kDa and their resistance to florfenicol substantially increased. It is speculated that the outer membrane protein changes might correlate with decreased susceptibility to florfenicol in the three strains. Some strains which showed completely identical OMP types had a little difference in their resistance to fluoroquinolones, indicating that there might be other factors that were involved in the antimicrobial resistance of A. hydrophila.