Antimicrobial activity of polyphenolic compounds from Spirulina against food-borne bacterial pathogens

Food-borne drug-resistant bacteria have adverse impacts on both food manufacturers and consumers. Disillusionment with the efficacy of current preservatives and antibiotics for controlling food-borne pathogens, especially drug-resistant bacteria, has led to a search for safer alternatives from natural sources. Spirulina have been recognized as a food supplement, natural colorant, and enriched source of bioactive secondary metabolites. The main objectives of this study were to isolate polyphenolic compounds from Spirulina and analyze their antibacterial potential against drug-resistant food-borne bacterial pathogens. We found that fraction B of methanol extract contained a high quantity of polyphenols exhibiting broad spectrum antimicrobial effects against drug-resistant food-borne bacterial pathogens. Potential secondary metabolites, such as benzophenone, dihydro-methyl-phenylacridine, carbanilic acid, dinitrobenzoate, propanediamine, isoquinoline, piperidin, oxazolidin, and pyrrolidine, were identified by gas chromatography and mass spectrophotometry (GCMS). These metabolites are active against both gram-positive and gram-negative pathogens. Our work suggests that phenolic compounds from Spirulina provide a natural and sustainable source of food preservatives for future use.     

Antifungal activity of Moroccan medicinal plants against citrus sour rot agent Geotrichum candidum

Aims: The aim of this work was to find an alternative to the chemical fungicides currently used in the control of Geotrichum candidum, the causal agent of citrus sour rot. Methods and Results: Minimal inhibition concentrations (MIC) and minimal fungicidal concentrations (MFC) were determined using agar dilution method. The methanol extracts of Cistus villosus, Ceratonia siliqua and Halimium umbellatum exhibited strong antifungal activity with MIC values ranged between 0·156 and 1·25 mg ml^?1, and MFC values ranged between 2·5 and 5 mg ml^?1. Incidence of sour rot was lowered to 0·00, 3·33 and 11·66% when mandarin fruit was treated with C. villosus, C. siliqua and H. umbellatum methanol extracts at 50 mg ml^?1, respectively, compared with 95% in the control. Conclusions: Cistus villosus, C. siliqua and H. umbellatum methanol extracts successfully reduced the disease incidence caused by G. candidum, and no phytotoxic effects were recorded on citrus fruit. Significance and Impact of the Study: These findings suggest that C. villosus, C. siliqua and H. umbellatum plants may be useful and effective agents for control of citrus sour rot. Such natural products therefore represent a sustainable alternative to the use of synthetic fungicides.     

Morphological and molecular characterisation of Lasiodiplodia theobromae from various banana cultivars causing crown rot disease in fruits

Twelve isolates of Lasiodiplodia theobromae were collected from 12 different commercial banana cultivars belonging to various banana growing regions of Tamil Nadu, India. The results revealed that most of the L. theobromae isolates exhibited significant differences in morphology, colour and spore size. However, all 12 isolates took 3–4 days invariably to cover the 90 mm Petri plates. Among the 12 isolates, the isolates LT3a from Robusta and LT10c from dwarf Cavendish produced abundant pycnidia. These isolates were also analysed through Random amplified polymorphic DNA (RAPD) markers for genetic diversity. Polymerase chain reaction amplification of total genomic DNA of the 12 isolates of L. theobromae with 14 random primers generated numerous polymorphisms among the isolates, while many of the intense bands were shared among majority of the isolates. Cluster analysis also indicated a high degree of genetic variability within L. theobromae isolates from different banana cultivars. The 12 isolates were separated into three clusters namely A, B and C. The maximum similarity index of 80% was recorded between the isolates LT7d from Karpuravalli and LT10c from dwarf Cavendish. The least similarity index of 29.41% was recorded with the isolates LT1f isolated from Poovan and LT5a isolated from Virupakshi. Within the species of L. theobromae the genetic variability was high and it underlines and validates existence of significant intra-specific diversity in isolates of L. theobromae infecting different banana cultivars.     

Fungicidal activity of essential oils of Cinnamomum zeylanicum (L.) and Syzygium aromaticum (L.) Merr et L.M. Perry against crown rot and anthracnose pathogens isolated from banana

AIMS: To develop a post-harvest treatment system against post-harvest fungal pathogens of banana using natural products. METHODS AND RESULTS: Colletotrichum musae was isolated and identified as the causative agent responsible for anthracnose peel blemishes while three fungi, namely Lasiodiplodia theobromae, C. musae and Fusarium proliferatum, were identified as causative agents responsible for crown rot. During the liquid bioassay, cinnamon [Cinnamomum zeylanicum (L.)] leaf, bark and clove [Syzygium aromaticum (L.)] oils were tested against the anthracnose and crown rot pathogens. The test oils were fungistatic and fungicidal against the test pathogens within a range of 0.03-0.11% (v/v). CONCLUSIONS: Cinnamon and clove essential oils could be used as antifungal agents to manage post harvest fungal diseases of banana. SIGNIFICANCE AND IMPACT OF THE STUDY: Cinnamon and clove essential oil could be used as alternative post-harvest treatments on banana. Banana treated with essential oil is chemically safe and acceptable to consumers. Benomyl (Benlate), which is currently used to manage fungal pathogens, can cause adverse health effects and could be replaced with volatile essential oils.     

Efficacy of leaf extracts of some medicinal plants against disease development in banana

Treatment of infested banana fruits by aqueous leaf extracts of some medicinal plants, viz. Calotropis procera (Ait) R.Br., Vitex negundo Linn., Lantana camara Linn., Azadirachta indica Linn., Ficus religiosa Linn., Ocimum sanctum Lin., Thuja orientalis Linn., Argemone mexicana Linn., Achyranthes aspera Linn., Datura fastuosa Linn. and Ricinus communis Linn. exhibited considerable control of disease development. Of 11 leaf extracts, those of A. indica and O. sanctum were most effective, and showed minimum per cent loss in fruit weight. The appearance of the first disease symptom was also delayed in treated fruits as compared with untreated ones.     

Establishment of embryogenic cell suspension cultures and Agrobacterium-mediated transformation in an important Cavendish banana cv. Robusta (AAA)

Establishment of an efficient protocol for regeneration and genetic transformation is required in banana for the incorporation of useful traits. Therefore an efficient method has been developed for somatic embryogenesis, plant regeneration and transformation of Cavendish banana cultivar Robusta (AAA). Embryogenic cell suspension culture (ECS) was established using immature male flowers. Percentage appearance of embryogenic callus and distinct globular embryos was 10.3 and 11.1, respectively. ECS obtained was cocultivated under different cocultivation conditions with Agrobacterium tumefaciens strain EHA105 harboring pCAMBIA 1301 plant expression vector. Up to 30 transgenic plants/50 mg settled cell volume (SCV) was obtained with cocultivation in semisolid medium whereas no transgenics could be obtained with parallel experiments carried out in liquid medium. Histochemical GUS assay in different tissues of putatively transformed plants demonstrated expression of uidA gene. Among the putatively transformed plants obtained, a set of 4 were confirmed by PCR analysis and stable integration of the transgene by Southern analysis. GUS specific activity measured by a MUG (4-methylumbelliferyl-β-d-glucuronide) based flourometric assay revealed increase in transient GUS expression in semisolid as well as liquid cocultivation with centrifugation. This is the first report showing somatic embryogenesis and Agrobacterium tumefaciens mediated transformation using embryogenic cell suspension cultures in an important Cavendish banana cultivar Robusta. The present protocol will make possible agronomic improvement of this important commercially grown cultivar by introduction of disease resistance characteristics and antisense-mediated delayed fruit ripening strategies. Further, it will also assist in functional characterization of new gene or promoter elements isolated from this or other cultivars of banana.     

Antimicrobial activity of Thymus longicaulis C. Presl essential oil against respiratory pathogens

Thymus longicaulis C. Presl is a small aromatic perennial herb used as a traditional remedy for cold, flu and cough. Composition of the essential oil of T. longicaulis from Croatia and its in vitro antimicrobial activity against the most common respiratory pathogens were evaluated. The yield of essential oil obtained by hydrodistillation from aerial plant parts was 1.2%. According to the GC-MS analysis, a total of forty one compounds (99%) were identified. Thymol (46.3%), ??-terpinene (16.2%), thymyl methyl ether (11.4%), and p-cymene (9.4%) were the main components. Antimicrobial activity of the essential oil against six clinically isolated bacterial and yeast strains was determined using standard disc agar diffusion method and microdilution broth assay. The essential oil exhibited antimicrobial activity towards all tested respiratory pathogens. The most sensitive strains were Haemophilus influenzae and Streptococcus pneumoniae(MIC=0.78 mg/mL), while Staphylococcus aureus was the most resistant (MIC>25.00 mg/mL). Our results indicate that T. longicaulis essential oil could be effective against clinically relevant respiratory pathogens which have the ability to develop resistance to antimicrobial drugs.     

Antimicrobial activities of 26 Yunnan plants extracts against clinical multi-drug resistant pathogens in vitro

研究26种云南滇东南产植物80％乙醇提取物的体外抗耐药菌作用.制备植物的醇提物,用琼脂打孔法对其进行抗菌活性的筛选.通过微量稀释法测定提取物的最低抑菌浓度[minimum inhibitory concentration(MIC)]、最低杀细菌浓度[minimum bactericidal concentration(MBC)]及最低杀真菌浓度[minimum fungicidal concentration(MFC)].结果表明:26种提取物中有9种对金黄色葡萄球菌(简称金葡菌)、大肠埃希菌、铜绿假单胞菌、白色念珠菌有不同程度的抑制活性;其中以红花荷、马蹄参、百花贝母兰、二色大包兰和光序肉实树提取物对耐甲氧西林金葡菌的抑菌作用最强,其MIC/MBC(mg/L)范围分别为512/2048～＞2048,512/2048～＞2048,256-512/2048～＞2048,512～1024/1024～＞2048和512～1024/＞2048.另外,肋果茶提取物对铜绿假单胞菌及白色念珠菌有较好的抑制作用,其对铜绿假单胞菌及其耐药菌株的MIC/MBC为512～2048/＞2048mg/L;对白色念珠菌及其耐药菌株的MIC/MFC均为2048/＞2048 mg/L.26种植物提取物对大肠埃希菌的抑制作用均较弱.     

26种滇产植物提取物的体外抗耐药菌作用研究

研究26种云南滇东南产植物80%乙醇提取物的体外抗耐药菌作用。制备植物的醇提物,用琼脂打孔法对其进行抗菌活性的筛选。通过微量稀释法测定提取物的最低抑菌浓度[minimum inhibitory concentration(MIC)]、最低杀细菌浓度[minimum bactericidal concentration(MBC)]及最低杀真菌浓度[minimum fungicidal concentration(MFC)]。结果表明:26种提取物中有9种对金黄色葡萄球菌(简称金葡菌)、大肠埃希菌、铜绿假单胞菌、白色念珠菌有不同程度的抑制活性;其中以红花荷、马蹄参、百花贝母兰、二色大包兰和光序肉实树提取物对耐甲氧西林金葡菌的抑菌作用最强,其MIC/MBC(mg/L)范围分别为512/2048～>2048,512/2048～>2048,256-512/2048～>2048,512～1024/1024～>2048和512～1024/>2048。另外,肋果茶提取物对铜绿假单胞菌及白色念珠菌有较好的抑制作用,其对铜绿假单胞菌及其耐药菌株的MIC/MBC为512～2048/>2048mg/L;对白色念珠菌及其耐药菌株的MIC/MFC均为2048/>2048mg/L。26种植物提取物对大肠埃希菌的抑制作用均较弱。     

Insecticidal activity of five Chinese medicinal plants against Plutella xylostella L. larvae

Twenty-five extracts of five Chinese medicinal plant species were screened for insecticidal activity against the diamondback moth, Plutella xylostella L., larvae by a leaf dipping bioassay method. The ethyl acetate, ethanol, and acetone extracts of Veratrum nigrum L. root and rhizome, the ethyl acetate extract of Phytolacca americana L. root, and the petroleum ether extract of Pseudolarix kaempferi Gord. root bark were effective against P. xylostella. Among them, the ethyl acetate extract of V. nigrum showed the strongest insecticidal activity against the second and third instar larvae of P. xylostella, with LC₅₀ values of 225 and 335 ppm, respectively. The other extracts gave little or no insecticidal activity against P. xylostella. These results indicated that V. nigrum may be a promising naturally occurring agent for P. xylostella larval control, and that the organic solvent used for plant extraction can affect its insecticidal activity.     

Evaluation of in vitro antifungal activity of medicinal plants against phytopathogenic fungi

Fourteen medicinal plants belonging to 13 families were collected and extracted with petroleum ether (PE), chloroform, methanol and water to yield 60 crude extracts. Using agar diffusion method, these extracts were evaluated for antifungal activity on the growth of five phytopathogenic fungi. Among all the extracts tested, PE, chloroform and methanol extracts of Piper betle L. and PE and chloroform extracts of Allamanda cathartica exhibited promising antifungal activity. Minimum inhibitory concentration (MIC) values of the above promising extracts were determined using broth dilution technique and observed that chloroform extract of P. betle L. exhibited the least MIC value ranging from 280 to 1130 μg ml^?1. In this study, we report chloroform extract of P. betle L. to be thermally stable even when steam sterilised for the first time and that it could be stored at 4°C with almost no change in its activity for a period of 180 days.     

甘草提取物对四种牙周常见致病菌的抑菌作用

目的 体外评价甘草提取物对牙龈卟啉单胞菌、中间普氏菌、具核梭杆菌和伴放线放线杆菌四种牙周常见致病菌的抑制效果。方法 以牙龈卟啉单胞菌、中间普氏菌、具核梭杆菌和伴放线放线杆菌四种牙周常见致病菌作为供试菌,采用液体稀释法,考察甘草提取物对这四种细菌的最小抑菌浓度（MIC）和最小杀菌浓度（MBC）;并采用不同浓度的甘草提取物溶液,绘制甘草提取物对四种牙周致病菌的时间‒杀菌曲线。结果 甘草提取物对牙龈卟啉单胞菌、中间普氏菌、具核梭杆菌和伴放线放线杆菌的MIC值分别为1.50、1.50、0.75和1.50 mg/mL,MBC值分别为6、3、3和3 mg/mL。当甘草提取物达到对四种细菌的MBC值时,对于牙龈卟啉单胞菌、中间普氏菌、伴放线放线杆菌可在2 h后可达到杀菌效果,对于具核梭杆菌可在4 h后达到杀菌效果。结论 甘草提取物对以上四种牙周常见致病菌具有良好的抑菌及杀菌作用。     

Antimicrobial, antitumor and antileishmania screening of medicinal plants from Guinea-Bissau.

Following an ethnobotanical search carried out in Guinea-Bissau, eighteen extracts derived from sixteen medicinal species were screened for antimicrobial, antitumor and antileishmania activity. Significant antitumor activity was found for Holarrhena floribunda against KB (squamous carcinoma), SK-Mel 28 (melanoma), A 549 (lung carcinoma) and MDA-MB 231 (mamma carcinoma) cell lines, with corresponding IC50 values of 7.9, 9.0, 3.4 and 9.9 micrograms/ml. Khaya senegalensis and Anthostema senegalense exhibited a significant activity against Leishmania donovani with IC50 values of 9.8 and 9.1 micrograms/ml, respectively. Most of the extracts showed week or moderate antibacterial and antifungal activity, with MIC values in the range 0.25-1.0 mg/ml. Active extracts were submitted to bioassay-guided fractionation, and the IC50 and MIC of the active fractions were determined.     

Antimicrobial activity of chemically and biologically synthesized silver nanoparticles against some fish pathogens

Pathogens isolated from fish appear to possess considerable antimicrobial resistance and represent a problem for the economy and public health. Natural antimicrobial substitutes to traditional antibiotics represent an essential tool in the fight against antibiotic resistance. Nanotechnology has shown considerable potential in different research fields, and the antimicrobial properties of silver nanoparticles are known. Silver has been used for medical purposes since ancient times because of its bactericidal properties, and the highly reactive surfaces of silver nanoparticles (AgNPs) indicate that they might have a function in antimicrobial applications. This work aimed to study the antimicrobial properties of biologically produced AgNPs from Origanum vulgare leaves compared to chemically produced AgNPs. Both types were characterized by UV–vis spectrophotometry, TEM, and dynamic light scattering and tested against three bacterial strains (Streptococcus agalactiae, and Aeromonas hydrophila, both isolated from Nile tilapia and Vibrio alginolyticus, isolated from sea bass) and three fungal strains (Aspergillus flavus, Fusarium moniliforme, and Candida albicans, all isolated from Nile tilapia). Disk diffusion test and evaluation of ultrastructure changes of tested microorganisms treated with AgNPs by transmission electron microscopy were performed. Moreover, the hemolytic properties of AgNPs were studied on chicken and goat red blood cells. The results obtained declare that the green biological production of silver nanoparticles is safer and more effective than the chemical one; moreover, AgNPs have interesting dose-dependent antimicrobial properties, with better results for biologically produced ones; their effectiveness against tested bacterial and fungal strains opens the way to their use to limit fish diseases, increase economy and improve human health.     

Antimicrobial activity of the marine alkaloids haminol and pulo'upone and related compounds

The marine alkaloids haminol A, haminol B and pulo'upone as well as 17 related compounds (twelve 2-substituted pyridine derivatives, four 3-substituted ones and one analogue of the bicyclic terminus of pulo'upone) were tested for antimicrobial activity against a panel of six microbes (Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis, Candida albicans and Saccharomyces cerevisiae) using the paper disc agar diffusion method. Six compounds were tested also against the mold Aspergillus niger. Some of the compounds displayed noteworthy antimicrobial activity, only one congener being completely devoid of activity. Nearly all compounds had activity against B. cereus and S. epidermidis. The growth of E. coli, C albicans and S. cerevisiae was also distinctly inhibited by many compounds. In contrast, most compounds were inactive or had minimal activity against P. aeruginosa. Interestingly, most of the compounds tested against the opportunistic pathogen A. niger were active, one of them having noteworthy inhibitory potency.     

Antimicrobial activity of silymarin mediated zinc oxide and hydroxy apatite nanoparticles against oral pathogens

The nanoparticles such as hydroxyapatite, zinc oxide, titanium dioxide and zirconium nanoparticles have application in dentistry. Therefore, it is of interest to document the antimicrobial activity of silymarin mediated zinc oxide and hydroxy apatite nanoparticles against oral pathogens. Hence, we synthesized hydroxyapatie and zinc oxide nanoparticles with silymarin and characterized by UV-visible spectrophotometer. Data shows that silymarin mediated HAP and ZnO nanoparticles have antimicrobial activity against oral pathogens such as Pseudomonas sp, Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis and Candida albicans.     

Construction of banana bunchy top nanovirus-DNA-3 encoding the coat protein gene and its introducing into banana plants cv. Williams

Banana (Musa sp.) is considered as one of the most important fruit crops worldwide as well as in Egypt. The main goal of this study was to construct the open reading frame (ORF) of banana bunchy top nanovirus (BBTV)-DNA-3 that encodes the viral coat protein (cp) gene for banana transformation. The previously sequenced BBTV-G-DNA-3-ORF that cloned into plasmid pH1 was used as a template for PCR amplification using two specific primers containing Bam H1 site. A new plasmid called pRHA1 containing the amplified ORF under the control of maize polyubiquitin (ubi) promoter was created. The bar gene (herbicide-resistance gene as a selectable marker) cassette (bar gene, Cauliflower mosaic caulimovirus (CaMV) 35S promoter and nos terminator) was released from plasmid pAB6 using Hind III-digestion and subcloned into the Hind III-digested plasmid pRHA1 to create the plasmid pRHA2 via the microprojectile bombardment transformation system. The plasmid pRHA2 was successfully introduced in the applied banana cultivar. Leaf painting test was conducted to confirm the expression of the bar gene in the putative transformed banana lines. The presence and expression of BBTV-G-cp gene were also detected using some molecular (polymerase chain reaction and dot blot using a cold DNA probe) and serological (ELISA and western blot) techniques, respectively, in the obtained transgenic banana lines.     

Antiviral activity of berberine and related compounds against human cytomegalovirus

Berberine chloride (1) and the structurally related compounds were assessed for the anti-human cytomegalovirus (HCMV) activity using the plaque assay. The anti-HCMV activity (IC(50) 0.68 microM) of 1 was equivalent to that (IC(50) 0.91 microM) of ganciclovir (GCV). The mechanism of action by which 1 inhibits the replication of HCMV is presumed to be different from that of GCV; 1 would interfere with intracellular events after virus penetration into the host cells and before viral DNA synthesis.