Isolation of Bacteria with Antifungal Activity against the Phytopathogenic Fungi Stenocarpella maydis and Stenocarpella macrospora

Stenocarpella maydis and Stenocarpella macrospora are the causal agents of ear rot in corn, which is one of the most destructive diseases in this crop worldwide. These fungi are important mycotoxin producers that cause different pathologies in farmed animals and represent an important risk for humans. In this work, 160 strains were isolated from soil of corn crops of which 10 showed antifungal activity against these phytopathogens, which, were identified as: Bacillus subtilis, Pseudomonas spp., Pseudomonas fluorescens, and Pantoea agglomerans by sequencing of 16S rRNA gene and the phylogenetic analysis. From cultures of each strain, extracellular filtrates were obtained and assayed to determine antifungal activity. The best filtrates were obtained in the stationary phase of B. subtilis cultures that were stable to the temperature and extreme pH values; in addition they did not show a cytotoxicity effect against brine shrimp and inhibited germination of conidia. The bacteria described in this work have the potential to be used in the control of white ear rot disease.     

20种植物提取物对植物病原细菌的抑制活性

[方法]以水稻白叶枯病菌(Xanthomonas oryzae pv.oryzae)、桃树细菌性穿孔病菌[Xanthomonus campestrispv.pruni(Smith) Dye.]为供试菌,对20种植物提取液进行抑菌活性筛选;采用液-液萃取法初步分离石榴皮、五倍子的抑菌活性成分.[结果]石榴皮、五倍子对2种病原细菌抑制效果最好;石榴皮的抑菌活性成分主要存在于正丁醇萃取层,五倍子乙酸乙酯萃取物的抑菌活性成分较好.[结论]石榴皮对2种病原细菌的抑制活性最好,其正丁醇萃取物是下一步研究的重点.     

黄花蒿(Artemisia annua L.)提取物对两种病原真菌的生物活性

以石油醚Ⅰ(30℃～60℃)、石油醚Ⅱ(60℃～90℃)、乙醇和丙酮等4种溶剂对黄花蒿的根、茎、叶进行初步提取,采用生长速率法测定不同提取物对玉米小斑病菌、棉花枯萎病菌的抑菌活性。结果表明,黄花蒿提取物对玉米小斑病菌的生物活性好于棉花枯萎病菌;黄花蒿叶的提取物抑菌效果最好,根的抑菌效果最差;叶的石油醚(60℃～90℃)提取物对玉米小斑病菌EC50为156.32mg/L;而叶的丙酮提取物对玉米小斑病菌的EC50为82.37mg/L。     

Remodeling of Lipid A in Pseudomonas syringae pv. phaseolicola In Vitro

Pseudomonas species infect a variety of organisms, including mammals and plants. Mammalian pathogens of the Pseudomonas family modify their lipid A during host entry to evade immune responses and to create an effective barrier against different environments, for example by removal of primary acyl chains, addition of phosphoethanolamine (P-EtN) to primary phosphates, and hydroxylation of secondary acyl chains. For Pseudomonas syringae pv. phaseolicola (Pph) 1448A, an economically important pathogen of beans, we observed similar lipid A modifications by mass spectrometric analysis. Therefore, we investigated predicted proteomes of various plant-associated Pseudomonas spp. for putative lipid A-modifying proteins using the well-studied mammalian pathogen Pseudomonas aeruginosa as a reference. We generated isogenic mutant strains of candidate genes and analyzed their lipid A. We show that the function of PagL, LpxO, and EptA is generally conserved in Pph 1448A. PagL-mediated de-acylation occurs at the distal glucosamine, whereas LpxO hydroxylates the secondary acyl chain on the distal glucosamine. The addition of P-EtN catalyzed by EptA occurs at both phosphates of lipid A. Our study characterizes lipid A modifications in vitro and provides a useful set of mutant strains relevant for further functional studies on lipid A modifications in Pph 1448A.     

Antifungal Activity of Essential Oil and Extracts of <Emphasis Type="Italic">Piper chaba</Emphasis> Hunter Against Phytopathogenic Fungi

The efficacy of the essential oil and various leaf extracts of Piper chaba Hunter was evaluated for controlling the growth of some important phytopathogenic fungi. The hydrodistilled essential oil was analysed by GC–MS. Fifty-three compounds representing 95.1% of the total oil were identified, of which α-humulene (16.4%), caryophyllene oxide (12.2%), viridiflorol (8.1%), globulol (7.4%), β-selinene (7.1%), spathulenol (6.2%), (E)-nerolidol (5.1%), linalool (4.5%) and 3-pentanol (3.5%) were the major compounds. The oil (1,000 ppm) and organic extracts (1,500 ppm) of P. chaba Hunter displayed a great potential of antifungal activity as a growth inhibition against the tested phytopathogenic fungi, Fusarium oxysporum KACC 41083, Phytophthora capsici KACC 40157, Colletotrichum capsici KACC 410978, Fusarium solani KACC 41092 and Rhizoctonia solani KACC 40111 in the range of 55.1–70.3% and minimum inhibitory concentration ranging from 125 to 500 μg/ml. The oil had also a strong detrimental effect on spore germination of all the tested fungi along with concentration as well as time-dependent kinetic inhibition of P. capsici. The results obtained from this study may contribute to the development of new antifungal agents to protect crops from fungal diseases.     

Image-Based In Vitro Screening Reveals the Trypanostatic Activity of Hydroxymethylnitrofurazone against Trypanosoma cruzi

Hydroxymethylnitrofurazone (NFOH) is a therapeutic candidate for Chagas disease (CD). It has negligible hepatotoxicity in a murine model compared to the front-line drug benznidazole (BZN). Here, using Trypanosoma cruzi strains that express bioluminescent and/or fluorescent reporter proteins, we further investigated the in vitro and in vivo activity of NFOH to define whether the compound is trypanocidal or trypanostatic. The in vitro activity was assessed by exploiting the fluorescent reporter strain using wash-out assays and real-time microscopy. For animal experimentation, BALB/c mice were inoculated with the bioluminescent reporter strain and assessed by highly sensitive in vivo and ex vivo imaging. Cyclophosphamide treatment was used to promote parasite relapse in the chronic stage of infection. Our data show that NFOH acts by a trypanostatic mechanism, and that it is more active than BZN in vitro against the infectious trypomastigote form of Trypanosoma cruzi. We also found that it is more effective at curing experimental infections in the chronic stage, compared with the acute stage, a feature that it shares with BZN. Therefore, given its reduced toxicity, enhanced anti-trypomastigote activity, and curative properties, NFOH can be considered as a potential therapeutic option for Chagas disease, perhaps in combination with other trypanocidal agents.     

Antiviral Activity of Two Marine Carotenoids against SARS-CoV-2 Virus Entry In Silico and In Vitro

The marine carotenoids fucoxanthin and siphonaxanthin are powerful antioxidants that are attracting focused attention to identify a variety of health benefits and industry applications. In this study, the binding energy of these carotenoids with the SARS-CoV-2 Spike-glycoprotein was predicted by molecular docking simulation, and their inhibitory activity was confirmed with SARS-CoV-2 pseudovirus on HEK293 cells overexpressing angiotensin-converting enzyme 2 (ACE2). Siphonaxanthin from Codium fragile showed significant antiviral activity with an IC50 of 87.4 μM against SARS-CoV-2 pseudovirus entry, while fucoxanthin from Undaria pinnatifida sporophyll did not. The acute toxicities were predicted to be relatively low, and pharmacokinetic predictions indicate GI absorption. Although further studies are needed to elucidate the inhibition of viral infection by siphonaxanthin, these results provide useful information in the application of these marine carotenoids for the treatment and prevention of COVID-19.     

Antibacterial Activity of Copper Nanoparticles against Xanthomonas campestris pv. vesicatoria in Tomato Plants

Copper-based bactericides have appeared as a new tool in crop protection and offer an effective solution to combat bacterial resistance. In this work, two copper nanoparticle products that were previously synthesized and evaluated against major bacterial and fungal pathogens were tested on their ability to control the bacterial spot disease of tomato. Growth of Xanthomonas campestris pv. vesicatoria, the causal agent of the disease, was significantly suppressed by both nanoparticles, which had superior function compared to conventional commercial formulations of copper. X-ray fluorescence spectrometry measurements in tomato leaves revealed that bioavailability of copper is superior in the case of nanoparticles compared to conventional formulations and is dependent on synthesis rather than size. This is the first report correlating bioavailability of copper to nanoparticle efficacy.     

Activity of Oritavancin and Its Synergy with Other Antibiotics against Mycobacterium abscessus Infection In Vitro and In Vivo

Background: Pulmonary disease caused by Mycobacterium abscessus (M. abscessus) spreads around the world, and this disease is extremely difficult to treat due to intrinsic and acquired resistance of the pathogen to many approved antibiotics. M. abscessus is regarded as one of the most drug-resistant mycobacteria, with very limited therapeutic options. Methods: Whole-cell growth inhibition assays was performed to screen and identify novel inhibitors. The IC₅₀ of the target compounds were tested against THP-1 cells was determined to calculate the selectivity index, and then time–kill kinetics assay was performed against M. abscessus. Subsequently, the synergy of oritavancin with other antibiotics was evaluated by using checkerboard method. Finally, in vivo efficacy was determined in an immunosuppressive murine model simulating M. abscessus infection. Results: We have identified oritavancin as a potential agent against M. abscessus. Oritavancin exhibited time-concentration dependent bactericidal activity against M. abscessus and it also displayed synergy with clarithromycin, tigecycline, cefoxitin, moxifloxacin, and meropenem in vitro. Additionally, oritavancin had bactericidal effect on intracellular M. abscessus. Oritavancin significantly reduced bacterial load in lung when it was used alone or in combination with cefoxitin and meropenem. Conclusions: Our in vitro and in vivo assay results indicated that oritavancin may be a viable treatment option against M. abscessus infection.     

植物病原物对杀菌剂的抗药性机制概述

就植物病原物对几大类杀菌剂的抗药性机制进行了介绍 ,强调了抗药性机制研究的重要性。     

苦槛蓝内生真菌不同培养条件代谢产物抗植物病原菌活性及LC-MS研究

利用滤纸片法和LC-MS分别对比了100多种苦槛蓝内生菌固液培养条件,高盐与无盐培养条件、单培养及共培养条件下抗菌活性及代谢产物成分的差异,报道了其中部分筛选出的改变相应培养条件后代谢产物活性显著增加及成分明显变化的菌株的实验数据.     

粤蛇葡萄及几种杀菌药剂对柑橘溃疡病菌的抑茵活性

采用带药平板涂布法测定了粤蛇葡萄甲醇提取物及5种杀菌药剂对柑橘溃疡病菌的抑菌活性.结果表明:粤蛇葡萄茎和叶提取物对柑橘溃疡病菌均有抑菌活性,最低抑菌质量浓度(MIC)分别为4、2 g/L;杀菌药剂60%甲硫·异菌脲WP、72%农用链霉素SP、硫酸铜晶体、53.8%氢氧化铜WG和64%吧霜灵WP对柑橘溃疡病菌的最低抑菌浓度分别为10、2.5、0.625、0.3125、0.1563 g/L.采用液-液萃取法对粤蛇葡萄的抑菌活性成分进行了初步分离,结合活性测定,发现抑菌活性成分主要存在于茎和叶提取物的乙酸乙酯萃取物中.     

Design and Pharmacodynamics of Recombinant Fungus Defensin NZL with Improved Activity against Staphylococcus hyicus In Vitro and In Vivo

Staphylococcus hyicus is recognized as a leading pathogen of exudative epidermitis in modern swine industry. Antimicrobial peptides are attractive candidates for development as potential therapeutics to combat the serious threats of the resistance of S. hyicus. In this study, a series of derivatives were designed based on the NZ2114 template with the aim of obtaining peptides with more potent antimicrobial activity through changing net positive charge or hydrophobicity. Among them, a variant designated as NZL was highly expressed in Pichia pastoris (P. pastoris) with total secreted protein of 1505 mg/L in a 5-L fermenter and exhibited enhanced antimicrobial activity relative to parent peptide NZ2114. Additionally, NZL could kill over 99% of S. hyicus NCTC10350 in vitro within 8 h and in Hacat cells. The results of membrane permeabilization assay, morphological observations, peptide localization assay showed that NZL had potent activity against S. hyicus, which maybe kill S. hyicus through action on the cell wall. NZL also showed an effective therapy in a mouse peritonitis model caused by S. hyicus, superior to NZ2114 or ceftriaxone. Overall, these findings can contribute to explore a novel potential candidate against S. hyicus infections.     

植物乳杆菌凝集素的纯化及其体外抗HIV活性

目的纯化植物乳杆菌(L.plantarum PCL)凝集素,并检测其体外抗HIV活性。方法采用Sephadex G-25凝胶脱盐层析和HITrap Q Sepharose XL离子交换层析纯化L.plantarum PCL凝集素;测定纯化产物的兔红细胞凝集素活性、纯度及荧光光谱;采用MTT法检测其对C8166淋巴细胞的毒性作用,以光镜检查其对HIV-1ⅢB诱导C8166细胞形成合胞体的抑制作用,并计算TI值。结果纯化的L.plantarum PCL凝集素经尿素-SDS-PAGE,可见相对分子质量约85000的单一条带;荧光光谱可见在380nm附近有一个发射强度较大的凝集素特征荧光发射峰;L.plantarum PCL凝集素对C8166细胞的IC50为2.239μg/ml,对HIV-1ⅢB诱导C8166细胞合胞体形成的EC50为0.079μg/ml,TI值为28.34。结论已成功纯化了L.plantarum PCL凝集素,并具有抗HIV活性。     

Synthesis and Antifungal Activity of Some Organotin(IV) Carboxylates

Six diorganotin(IV) carboxylates prepared by reacting diorganotin(IV) dichlorides with the respective silver carboxylate have been tested for antifungal activity against Aspergillus. niger, Aspergilluus flavus and Pencillium. citrinum in Sabourand dextrose broth. The compounds generally exhibit greater fungitoxicity than the diorganotin(IV) dichlorides and the carboxylic acids from which they were synthesized. In keeping with the generally accepted notion that the organotin moiety plays an important role in deciding the antifungal activity of an organotin compound, the diphenyltin(IV) compounds were more active than their di-n-butyltin(IV) analogues. However, the order of increasing fungitoxicity of the compounds parallels that of the uncomplexed carboxylic acids. The implications of the results are discussed.     

Chemical Composition and in Vitro Antifungal Activity Screening of the Allium ursinum L. (Liliaceae)

The objective of the study was to summarize the methods for isolating and identifying natural sulfur compounds from Allium ursinum (ramson) and to discuss the active constituents with regard to antifungal action. Using chromatographic techniques, the active constituents were isolated and subsequently identified. Analyses by high-performance liquid chromatography (HPLC) suggested that these compounds were sulfur constituents, with a characteristic absorbance at 250 nm. Gas chromatography-mass spectrometry (GC-MS) analyses allowed the chemical structures of the isolated constituents to be postulated. We adopted the same methods to identify the health-giving profiling of ramsons and the effects are thought to be primarily derived from the presence and breakdown of the alk(en)ylcysteine sulphoxide, alliin and its subsequent breakdown to allicin (sulfur-compounds of ramson) in connection with antifungal action. The aim of the study was the characterization of the chemical composition of ramsons and the testing of the action of the in vitro extracts, on different strains of Candida albicans. The main goal was to highlight the most efficient extracts of Allium ursinum that can provide long-term antifungal activity without remissions. The extracts from Allium ursinum plants, inhibited growth of Candida spp. cells at concentrations ranging from 0.5 to 4.0 mg/mL, while that of adherent cells at concentrations ranging from 1.0 to > 4.0 mg/mL, depending on the yeast and plant species.     

Anti-Inflammatory Activity of Mulberry Leaf Flavonoids In Vitro and In Vivo

Mulberry (Morus alba L.) is a flowering tree traditionally used in Chinese herbal medicine. Mulberry leaf flavonoids (MLFs) have been reported to exert important anti-inflammatory and antioxidant properties. The purpose of this study was to select the MLF with the best anti-inflammatory and antioxidative activities from MLFs eluted by different ethanol concentrations (30%, 50%, and 75%) and explore its pharmacological properties. Three types of MLFs inhibited the production of nitric oxide (NO), prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and inflammatory cytokines in lipopolysaccharide (LPS)-induced RAW 264.7 cells. All MLFs boosted the antioxidative capacity by decreasing the reactive oxygen species (ROS) production and the scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals and improving the metal ion chelating activity and reducing power. The results revealed that the MLFs eluted by 30% ethanol exhibited the best anti-inflammatory and antioxidative activities. A nontargeted metabolomic analysis was used to analyze 24 types of differential flavonoids between the MLFs. Quercetin, kaempferol, and their derivatives in 30%MLF were more abundant than the other two MLFs. Furthermore, we evaluated the pharmacological activities of 30%MLF in dextran sodium sulfate (DSS)-induced ulcerative colitis (UC) mice. The 30%MLF could alleviate the clinical symptoms, reduce the secretion of inflammatory cytokines, and inhibit the activation of the inflammatory pathway in DSS-induced colitis mice. This study will provide valuable information for the development of MLFs eluted by 30% ethanol as a functional food.     

Synthesis, In vitro Antifungal and Antitumour Activity of Some Triorganotin(IV) N,C,N-Chelates

The in vitro antifungal activity of compounds 1-3 ({[(CH3)2NCH2]2C6H3}R2SnX; (where X=Cl, R=n-Bu for 1, X=Br, R=n-Bu for 2 and x=PF6, R=n=Bu for 3)) was estimated with the help of a modified microdilution format of the M27-A guidelines and was compared with in vitro activity of their diphenyltin(IV) analogues 4 and 5 (where X=Br, R=Ph for 4 and X=PF6, R=Ph for 5), and of drugs currently in clinical use (ketoconazole, fluconazole and amphotericin B). It was found that in coordinating solvents the more soluble derivative 2 is less active than the phenyl one (4), and compounds 1 and 3 are even inactive.In this paper, the in vitro antitumour activity of ionic diphenyltin(IV) complexes 4 and 5 against seven tumoural cell lines of human origin is also reported. The preparation and characterization (H1, C13 and Sn119 NMR spectroscopy and electrospray ionization mass spectrometry) of the novel compound 3 is mentioned too.     

化香果提取物体外抑菌活性研究

研究了化香果提取物对水产常见9种致病菌嗜水气单胞菌（Aeromonas hydrophila）、温和气单胞菌（Aeromonas sobria）、豚鼠气单胞菌（Aeromonas caviae）、维氏气单胞菌（Aeromonas veronii）、迟钝爱德华菌（Edwardsiella tarda）、副溶血弧菌（Vibrio parahaemolyticus）、鼠疫耶尔森氏菌（Yersinia pestis）、大肠杆菌（Escherichia coli）和金黄色葡萄球菌（Staphylococcus aureus）的抑菌活性。研究结果表明：化香果提取物对9种细菌均具有抑制作用,最小抑菌浓度为4.50 g/L。对每种细菌而言,抑菌效果随化香果提取物质量浓度的增加而增强,其中对大肠杆菌的抑制能力最强,IC₅₀值为0.035 g/L;金黄色葡萄球菌次之,IC₅₀值为0.053 g/L。对雌雄小鼠急性经口毒性试验结果表明化香果提取物对小鼠的急性经口半数致死量（LD₅₀值）分别为5 010和7 940 mg/kg体质量,表明化香果提取物属于实际无毒级。