益母草抗菌蛋白LJAMP2的表达及抗菌活性分析

LJAMP2是一种分离自益母草(Leonurus japonicus Houtt.)种子的非专一性脂转移蛋白类抗菌蛋白.将编码成熟的LJAMP2蛋白的基冈(GenBank Accession No.AY971513)插入到巴斯德毕赤酵母(Pichia pas-toris)表达载体pPIC9K中;重组质粒pPIC9K-LJAMP2经线性化后,用电转化法导入毕赤酵母菌株GS115(his-/mut+)中.转化子用含1.75 mg/mL G418的YPD平板筛选获得多拷贝Mut+转化子.将1%甲醇诱导LJAMP2表达后的发酵液,经进一步浓缩、凝胶过滤脱盐、CM FF阳离子交换色谱和反相HPLC进行纯化,获得纯度90%以上的LJAMP2菌株诱导24 h即有LJAMP2的表达,诱导168 h表达量达到最高;表达的LJAMP2蛋白对黑曲霉(Aspergillus niger)、玉米小斑病菌(Bipolaris maydis)、镰刀菌(Fusarium graminearum)真菌表现出抑制活性.     

An Experimental Study on the Treatment and Prevention of Shimao Zheng Fleece-Eating) in Sheep and Goats in the Haizi Area of Akesai County in China

A series of experimental trials on methods of treatment and prevention of shimao zheng fleece-eating) were carried out using various sulphates. The results confirmed the hypothesis that this is a local nutritional and metabolic disease, caused mainly by a sulphur deficiency, that could be prevented or cured with supplementary sulphur in the diet. In field studies, the clinical signs virtually disappeared within 14 days of the animals receiving medicated pellets. Over the same period, the concentrations of sulphur in the blood, liver and spleen returned to normal and the pathological changes in the skin improved. The condition of the controls became worse in each of these respects over this time. However, the pathological changes in the musculature were not alleviated by the treatment.     

Innate immunity and host defense peptides in veterinary medicine

Recent years have witnessed a surge in interest directed at innate immune mechanisms. Proper conceptualization of the key elements of innate immunity, however, is still a work in progress, because most research in immunology traditionally has been focused on components of the acquired immune response. The question of why an animal stays healthy in a world filled with many dangers is perhaps as interesting as why it sometimes surrenders to disease. Consequently, studies with an increased focus on inborn mechanisms of animal host defense may help further the development of appropriate preventative and therapeutic measures in veterinary medicine. Host defense peptides (HDPs) are central effector molecules of innate immunity, and are produced by virtually all living species throughout the plant and animal kingdoms. These gene-encoded peptides play a central role in multiple, clinically relevant disease processes. Imbalances in the expression of HDPs can lead to overt pathology in different organ systems and cell types in all species studied. In addition, HDPs are an ancient group of innate chemical protectors, which are now evaluated as model molecules for the development of novel natural antibiotics and immunoregulatory compounds. This review provides an overview of HDPs and is aimed at veterinary practitioners as well as basic researchers with an interest in comparative immunology involving small and large animal species.     

Multidrug-resistant E. coli and enterobacter extraintestinal infection in 37 dogs

BACKGROUND: Extraintestinal infections caused by multidrug-resistant (MDR) Escherichia coli and Enterobacter are becoming more common in veterinary medicine. OBJECTIVE: To generate hypotheses for risk factors for dogs acquiring extraintestinal infection caused by MDR E. coli and Enterobacter, describe antimicrobial resistance profiles and analyze treatment and clinical outcomes. ANIMALS: Thirty-seven dogs diagnosed with extraintestinal infection caused by MDR E. coli and Enterobacter spp. between October 1999 and June 2006. METHODS: Retrospective case series assembled from hospital records data, including clinical history before 1st MDR isolation and treatment outcome. Identity and antimicrobial susceptibility profiles were confirmed by standard microbiological techniques for 57 isolates. RESULTS: Most dogs had an underlying disease condition (97%), received prior antimicrobial treatment (87%), were hospitalized for >or =3 days (82%), and had a surgical intervention (57%). The urinary tract was the most common infection site (62%), and urinary catheterization, bladder stasis, or both were common among dogs (24%). Some dogs were treated with high doses of co-amoxyclavulanate (n = 14) and subsequently recovered even though the isolates showed in vitro resistance to this antimicrobial. Other dogs were successfully treated with chloramphenicol (n = 11) and imipenem (n = 2). CONCLUSION AND CLINICAL IMPORTANCE: Predisposing disease condition, any prior antimicrobial use rather than a specific class of antimicrobial, duration of hospitalization, and type of surgical procedure might be risk factors for acquiring MDR extraintestinal infections. Whereas culture and sensitivity results can indicate use of last-resort antimicrobials such as imipenem for MDR infections, some affected dogs can recover after administration of high doses of co-amoxyclavulanate.     

Antimicrobial-Resistant Enterococcus faecium and Enterococcus faecalis Isolated From Healthy Thoroughbred Racehorses in Japan

In this study, the occurrence of antimicrobial-resistant (AMR) enterococci was evaluated in Thoroughbred (TB) racehorses in Japan. Fecal samples were collected from 212 healthy TB racehorses at the Miho and Ritto Training Centers of the Japan Racing Association from March 2017 to August 2018. Isolation and identification were performed by enterococcus selective medium and confirmed to the species using MALDI-TOF MS. Enterococcus faecium and E. faecalis isolates were subjected to antimicrobial susceptibility test against 11 antimicrobials by minimum inhibitory concentration based on recommendation from Clinical and Laboratory Standards Institute guidelines. Among 583 enterococcus isolates, E. faecium and E. faecalis were identified for 48.2% (281/583) and 7.4% (43/583), respectively. One isolate that was representing E. faecium (153 isolates) and E. faecalis (31 isolates) from each sample was selected for antimicrobial susceptibility test. The highest rate of resistance for E. faecium isolates was observed against enrofloxacin (57.5%; 88/153), followed by streptomycin (32.0%; 49/153), kanamycin (18.3%; 28/153), gentamycin (5.9%; 9/153), erythromycin (5.9%; 9/153), and oxytetracycline (4.6%; 7/153). For E. faecium isolates, the highest resistance was observed against streptomycin (90.3%; 28/31), followed by kanamycin (41.9%; 13/31), gentamycin (29.0%; 9/31), lincomycin (9.7%; 3/31), oxytetracycline (6.5%; 2/31), erythromycin (6.5%; 2/31), tylosin (6.5%; 2/31), enrofloxacin (6.5%; 2/31), and chloramphenicol (3.2%; 1/31). The results indicated that enrofloxacin and aminoglycosides were highly resistant among tested antimicrobials. Continuous monitoring studies are useful to increase the awareness of the potential for AMR bacteria to arise from imprudent use of antimicrobials in TB racehorses in Japan.     

山东主要刺参养殖区幼参肠道抗生素耐药菌及耐药基因分布特征

为解析目前刺参(Apostichopus japonicus)苗种携带耐药菌及耐药基因现状,本研究从烟台、威海、青岛3个刺参主要养殖区的6家养殖场采集了幼参,对其肠道内容物中常用抗生素的耐药菌数量、占比和种类进行检测。利用荧光定量PCR技术,对样本中4类抗生素的7种耐药基因分布情况进行分析。结果显示,所检测的6个采样点中均有6种抗生素耐药菌的检出,从耐药菌占比来看,耐药菌占比最高的种类是乙酰甲喹、萘啶酸和四环素耐药菌,占比分别为0.05%~40.06%、2.16%~39.94%和0.06%~23.15%,氟苯尼考、庆大霉素和链霉素耐药菌的占比均不高,占比范围为0.01%~4.15%。由所分离到的98株耐药菌的鉴定结果可以看出,可培养的抗生素耐药菌分为4门5纲30属,主要集中在弧菌属、芽孢杆菌属和嗜冷杆菌属,占检出率的15.30%、13.27%和12.25%。基于属水平的不同抗生素耐药菌种类的分布统计结果显示,各采样点耐药菌种类差异较大,而且弧菌属、芽孢杆菌属和嗜冷杆菌属中均存在同一菌属中有耐多种抗生素的情况。对6个样本7种抗生素耐药基因的丰度检测结果显示,同一类抗生素的不同耐药基因的含量差异显著,除氨基糖苷类抗生素耐药基因aadA的相对拷贝数比例和链霉素以及庆大霉素耐药菌占比之间存在显著相关性(P<0.05)外,其他抗生素耐药基因的丰度与耐药菌占比之间相关性不显著(P>0.05)。研究表明,苗期刺参中存在一定的携带耐药菌和耐药基因的风险。     

昆虫免疫致敏响应研究综述

从昆虫免疫致敏识别、免疫致敏响应、免疫致敏响应分子机制、免疫致敏传递机制等方面对昆虫免疫致敏进行综述,同时对农业害虫抗性治理和农药合理运用等进行了展望。     

黄花蒿烟草醋液对三线镰刀菌生长的抑制作用

充分利用黄花蒿烟草秸秆制成秸秆混合醋液,并初步研究其对三线镰刀菌生长的影响。分离纯化真菌并对菌种进行传统形态学和分子学鉴定,利用干馏法制取黄花蒿烟草秸秆醋液,设置浓度梯度观察黄花蒿烟草秸秆醋液对三线镰刀菌2株菌的生长影响。鉴定获得的其中2株菌种均为三线镰刀菌,且在黄花蒿烟草秸秆醋液50%浓度下,生长抑制率的相对校正率近似为100%,达到完全被抑制的效果。研究结果为黄花蒿烟草秸秆潜在的利用价值研究及开发提供技术参考。     

Antimicrobial activity of bioactive compounds and leaf extracts in Jatropha tanjorensis

Objectives

Jatropha tanjorensis was investigated scientifically to generate evidence for the efficacies reported in traditional systems and the results are given here.

Methods

Different concentrations of the solvent extracts of leaves and four isolated compounds were tested against human pathogenic microorganisms such as gram-positive bacteria of Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermis, gram-negative bacteria of Aeromonas hydrophila, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Proteus vulgaris, Salmonella paratyphi, Salmonella paratyphi A, Vibrio alcaligenes, Vibrio cholerae and fungi of Aspergillus fumigatus, Candida albicans, Microsporum gypseum and Trichophyton rubrum by agar-well diffusion and disk diffusion methods.

Results

In agar-well diffusion method, maximum activity was recorded in a concentration-dependent manner. The extracts recorded activity against bacteria such as 17–26, 15–25 and 13–24 mm to methanol extract and 15–24, 14–23 and 12–22 mm to chloroform extract at 50, 25 and 12.5 mg/ml respectively and fungi such as 9–15 mm to A. fumigatus and 5–16 mm to T. rubrum. Maximum activity was 30–46, 27–43 and 17–40 mm to friedelin and 23–46, 28–44 and 18–41 mm to R (+) 4-hydroxy-2-pyrrolidinone against bacteria and 12–37, 8–34 and 31–33 mm to friedelin and 12–40, 11–35 and 10–33 mm to R (+) 4-hydroxy-2-pyrrolidinone against fungi at 10, 5 and 2.5 mg respectively.

Conclusions

The present study concludes that friedelin, β-amyrin, stigmasterol and R (+) 4-hydroxy-2-pyrrolidinone present in the methanol extract could be responsible for the broad spectrum of antimicrobial activity and provide scientific evidence.