Antibacterial and Antifungal Activities of Compounds Isolated from Gymnosporia Royleana (Celastraceae) / Actividades antibacterianas y antifúngicas de compuestos aislados de Gymnosporia Royleana (Celastraceae)

ABSTRACT Objective: In the present study, five new source compounds isolated from aerial parts of Gymnosporia royleana (G royleana) were screened for antibacterial and antifungal activities. Methods: Extraction from plant material was carried out using cold maceration technique. Isolation of pure compounds was accomplished through repeated column chromatography of different fractions obtained from crude extract and using silica gel as stationary phase. Their structures were established via advanced spectroscopic techniques along with the spectral data previously reported for these compounds. Dilution method was used for the evaluation of antimicrobial potential of the compounds against various microbial strains. Results: Among the tested compounds, Gymnosporin B displayed moderate antimicrobial activity against Escherichia Coli (E coli), Staphylococcus aureus (S aureus), Candida albicans (C albicans) and Aspergillus flavus (A flavus) [minimum inhibitory concentration (MIC) range; 32-64 μg/mL]. Similarly, Gymnosporin C also showed moderate activity against E coli and S aureus (MIC; 32 μg/mL each) as well as weak activity against C albicans and A flavus (MIC; 64 μg/mL each). However, Royaflavone showed moderate antibacterial activity against S aureus only (MIC; 32 μg/mL). Antimicrobial activity of the rest of the compounds was weak and negligible. Conclusion: The present study has provided fascinating results of antimicrobial activities of the isolated compounds. However, the broad antimicrobial spectrum of Gymnosporin B and Gymnosporin C demands for further exploration of these triterpenes, both on the basis of mechanism and quantitative structure-activity relationship.

RESUMEN Objetivo: En el presente estudio, cinco nuevos compuestos de origen aislados de partes aéreas de Gymnosporia royleana (G royleana) fueron tamizados en sus actividades antibacterianas y antifúngicas. Métodos: La extracción de material vegetal se realizó mediante la técnica de maceración en frío. El aislamiento de compuestos puros se logró a través de la cromatografía en columna repetida de diversas fracciones obtenidas del extracto crudo y usando gel de silicona como fase estacionaria. Sus estructuras fueron establecidas mediante técnicas espectroscópicas avanzadas junto con los datos espectrales previamente reportados para estos compuestos. El método de dilución fue usado para evaluar el potencial antimicrobiano de los compuestos contra diversas cepas microbianas. Resultados: Entre los compuestos sometidos a prueba, Gymnosporina B mostró una actividad antimicrobiana moderada contra Escherichia Coli (E coli), Staphylococcus aureus (S aureus), Candida Albicans (C albicans) y Aspergillus flavus (A flavus) [rango de concentración inhibitoria mínima (CIM); 32 - 64 μg/mL]. De manera similar, Gymnosporina C también mostró actividad moderada contra E coli y S aureus (CIM; 32 μg/mL cada uno) así como débil actividad frente a C albicans y A flavus (CIM; 64 μg/mL cada uno). Sin embargo, Royaflavone mostró actividad antibacteriana moderada sólo frente a S aureus (CIM; 32 μg/mL). La actividad antimicrobiana del resto de los compuestos fue débil e insignificante. Conclusión: El presente estudio ha proporcionado resultados interesantes acerca de las actividades antimicrobianas de los compuestos aislados. Sin embargo, el amplio espectro antimicrobiano de la Gymnosporina B y la Gymnosporina C exige una mayor exploración de estos triterpenos, tanto sobre la base del mecanismo como a partir de la relación cuantitativa estructura-actividad.

In vitro antifungal susceptibility of clinical and environmental isolates of Aspergillus fumigatus and Aspergillus flavus in Brazil

ABSTRACT The in vitro susceptibility of 105 clinical and environmental strains of Aspergillus fumigatus and Aspergillus flavus to antifungal drugs, such as amphotericin B, azoles, and echinocandins was evaluated by the broth microdilution method proposed by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Following the EUCAST-proposed breakpoints, 20% and 25% of the clinical and environmental isolates of A. fumigatus, respectively, were found to be resistant to itraconazole (Minimal Inhibitory Concentration, MIC > 2.0 mg/L). Voriconazole showed good activity against A. fumigatus and A. flavus strains, except for one clinical strain of A. fumigatus whose MIC was 4.0 mg/L. Posaconazole (≤0.25 mg/L) also showed appreciable activity against both species of Aspergillus, except for six A. fumigatus strains with relatively higher MICs (0.5 mg/L). The MICs for Amphotericin B ranged from 0.06 to 1.0 mg/L for A. fumigatus, but were much higher (0.5-8.0 mg/L) for A. flavus. Among the echinocandins, caspofungin showed a geometric mean of 0.078 and 0.113 against the clinical and environmental strains of A. flavus, respectively, but had elevated minimal effective concentrations (MECs) for seven of the A. fumigatus strains. Anidulafungin and micafungin exhibited considerable activity against both A. fumigatus and A. flavus isolates, except for one environmental isolate of A. fumigatus that showed an MEC of 1 mg/L to micafungin. Our study proposes that a detailed investigation of the antifungal susceptibility of the genus Aspergillus from different regions of Brazil is necessary for establishing a response profile against the different classes of antifungal agents used in the treatment of aspergillosis.

The use of chitosan in protecting wooden artifacts from damage by mold fungi

Background: Many buildings in Egypt e.g. museums, mosques and churches, do not possess controlled environments for minimizing the risks of damage of wooden artifacts due to the growth of fungi. Fungal damage usually appears as change in wood color, appearance of stains, and sometimes deformation of wooden surfaces. In this study we focused on the effect that some fungi exert on the properties of wooden artifacts and evaluated the effectiveness of different concentrations of chitosan on their protection against damage by mold fungi. Results: Samples were collected from different monuments and environments, and fungi growing on them were isolated and identified. The isolated Penicillium chrysogenum, Aspergillus flavus and /Aspergillus niger strains were used for the infestation of new pitch pine samples. The results revealed that the lightness of samples infected with any of the tested fungi decreased with increasing incubation times. XRD analysis showed that the crystallinity of incubated samples treated individually with the different concentrations of chitosan was lower than the crystallinity of infected samples. The crystallinity index measured by the first and the second method decreased after the first and second months but increased after the third and fourth months. This may due to the reducing of amorphous part by enzymes or acids produced by fungi in wooden samples. Conclusions: The growth of fungi on the treated wood samples decreased with increasing the concentration of chitosan. Hence, it was demonstrated that chitosan prevented fungal growth, and its use could be recommended for the protection of archeological wooden artifacts.

Copper induction and differential expression of laccase in Aspergillus flavus

Aspergillus flavus was isolated from soil and exhibited laccase activity under both constitutive and copper induced conditions. Spiking the medium with 1 mM copper sulfate resulted in an increase in the activity which reached 51.84 U/mL, a distinctive protein band was detected at 60 kDa. The extracellular enzyme was purified 81 fold using gel filtration chromatography and resulted in two different laccase fractions L1 and L2, the latter had a higher enzymatic activity which reached 79.57 U/mL and specific activity of 64.17 U/μg protein. The analysis of the spectrum of the L2 fraction showed a shoulder at 330 nm which is characteristic for T2/T3 copper centers; both copper and zinc were detected suggesting that this is an unconventional white laccase. Primers of laccase gene were designed and synthesized to recover specific gene from A. flavus. Sequence analysis indicated putative laccase (Genbank ID: JF683612) at the amino acid level suggesting a close identity to laccases from other genera containing the copper binding site. Decolorization of textile waste water under different conditions showed possible application in bioremediation within a short period of time. The effect of copper on A. flavus was concentration dependent.

Biosynthesis of silver nanoparticles using fresh extracts of Tridax procumbens Linn.

A simple and eco-friendly method for the synthesis of biogenic nanoparticles (NP’s) using an aqueous solution of T. procumbens fresh plant extract (leaf and stem) as a bioreductant is reported. The prepared biogenic nanoparticles were well characterized using U.V. visible spectroscopy, scanning electron microscopy, X-ray diffraction and Fourier-transform infrared spectroscopy. The particles were confirmed to be elemental crystal by X-ray diffraction. The potential applications of biosynthesized nanoparticles as antimicrobial (antibacterial and antifungal) against pathogens Escherichia coli, Vibrio cholerae, Aspergillus niger and Aspergillus flavus were demonstrated.

Drug susceptibility of aAspergillus flavus and A.fumigatus to itraconazole and amphotericin B

Aspergillosis is the most current causative agent of exogenous fungal nosocomial infection. This study was done to evaluate the drug susceptibility of Aspergillus flavus and A.fumigatus to itraconazole and amphotericin B. This Laboratory study was done on 25 Aspergillus fumigatus and 25 Aspergillus flavus species isolated from transplant's patients. Drug susceptibility test was done according to NCCLS M38-P document. Fungal suspensions of mentioned fungi were supplied with ranges 0.5-5x10[4] by spectrophotometer at 530 nm. Serial dilutions of drugs were supplied from 0.03125 to 16 microg/ml and MICs determined following 48h incubation at 35C. Obtained MICs ranges for Aspergillus fumigatus and Aspergillus flavus were 1-4 microg/ml and 0.5-4 micro g/ml for itraconazole, respectively while MICs ranges against Aspergillus fumigatus and Aspergillus flavus were 0.5-2 micro g/ml and 0.25-2 microg/ml for amphotericin B, respectively. Amphotericin B MICs were significantly lower than itraconazole [P<0.05]. Aspergillus flavus and A.fumigatus were susceptible to amphotericin B and itraconazole

Screening the antifungal activity of essential oils against decay fungi from palmyrah leaf handicrafts

BACKGROUND: The whitish tender leaves of Palmyrah are used for making handicrafts. The problem with these articles is discolouration with time and become more brittle due to fungal attack. This could be prevented by some protective coating. Instead of expensive and harmful chemicals we decided to test natural plant essential oils to control fungal attack. Palmyrah leaf article decay fungi were isolated from two different sites of Jaffna peninsula. In this investigation Antifungal Activity of different plant essential oils from neem (Azadirachta indica), castor (Ricinus communis), citronella (Cymbopogon sp) and camphor (Cinnamomum camphora) obtained from local market have been evaluated against isolated fungi. For screening of Antifungal activity, tests and controls were set to determine minimum inhibitory concentration (MIC) and Percentage of Growth Inhibition. RESULTS: Morphologically three different types of Palmyrah leaf decay fungi were isolated and characterized asAspergillus niger, Aspergillus flavus and Penicillium sp. Neem and castor oils have recorded no significant (0.05 > P) antifungal activity while citronella and camphor oils showed significantly different antifungal activity compared with control. Camphor oil and Citronella oil showed 100, 58.13% of average growth inhibition for A. niger. 96.38, 51.32% for A.flavus and 84.99, 72.76% forPenicillium sp respectively. Camphor oil showed highest percentage of growth inhibition at lowest minimum inhibitory concentration compared with citronella oil. Camphor oil was found to be highly antifungal and most effective against A niger, and A. flavus, compared with Penicillium sp and gave 100 percentage of growth inhibitions at 5, 1 and 15 ml/dl minimum inhibitory concentration respectively. CONCLUSION: Significantly higher broad-spectrum of antifungal activity was observed in camphor oil than other tested oils because it showed highest percentage of growth inhibition at lowest inhibitory concentration. Therefore it could be used for the development of new environmental friendly antifungal agent for the preservation of leafy handicrafts. Further formulation, field experiments are necessary to achieve this target.

In vitro activity of disinfectants against Aspergillus spp

Fungi of the Aspergillus genus are widespread and contaminate the environment. Thousands of conidia are released from each phialide and dispersed in the air every day. These fungi are considered important mycose-causing agents in hospitals. Due to this, research to determine prevalent fungi from the Aspergillus genus in hospital environments, and an adequate disinfection program in these areas is are needed. This study evaluated the susceptibility of Aspergillus spp. isolated from a veterinary environment against four disinfectants. Successive dilutions of disinfectants (log2) were used according to CLSI M38-A2 microdilution technique adapted to chemical agents against 18 isolates of this genus. After 72 hours of incubation, the Minimum Inhibiting Concentration and Minimum Fungicidal Concentration capable of inhibiting 50% and 90% of the isolates were determined. Chlorexidine-cetrimine, benzalconium chloride and a chlorophenol derivative proved to be effective against all isolates with a lower MIC than that suggested by the manufacturer, except for the A. flavus strain. Sodium hypochlorite was ineffective against three A. fumigatus, three A. flavus and one A. niger isolate. These results demonstrated that all studied disinfectants were effective against environmental isolates, with the exception of sodium hypochlorite, which showed lower effectiveness.

[Determine the minimum inhibitor concentration MIC of the essential oil of Rosmarinus officinalis on the growth of Aspergillus flavus]

Objective: Studing the antifungal effect of Rosemary oil on Aspergillus flavus. Methods: Eight Rosemarinus officinalis Samples were collected from the following areas: Qardahah, Tel Kalakh, Karah, Deir al-Ashayr, the Faculty of Agriculture, University of Damascus [two planted samples]. And two samples from Lebanon [Broumana]. Essential oil was extracted by salvin meek. and studied for its anti fungal growth on Aspergillus flavus by Determine the minimum inhibitor concentration MIC

Results: The MIC in 6 samples were either 40 or 80 in the remaining samples

Conclusion: This study shows that the oil of rosemary has an antifungal effect against Aspergillus flavus

antifungal, cytotoxic, antitermite and insecticidal activities of zizyphus jujube

Plants are very useful, self-generating machines, producing a variety of useful bioactive products. Keeping in view this idea, the crude methanolic extract and various fractions of Zizyphus jujuba were screened for antifungal, cytotoxic, antitermite and insecticidal activities. Low activity was shown by the crude methanolic extract [12%], n-hexane [9%], chloroform [20%] and ethyl acetate [14%] fraction against Penicillium notatum. Low activity was shown by the ft-hexane fraction against Aspergillus niger [10%] and Trichoderma harzianum [13%] and inactive against Aspergillus flavus, Fusarium oxysporum and Rhizopus stolonifer. The CHC1[3] fraction exhibited low activity of 10% against F. oxysporum while showing no activity against the rest of the test fungi. All the test samples were inactive against Rhizopus stolonifer. The crude methanolic extract was highly cytotoxic [73.33%] at the concentration of 1000 [microg/ml] while the rest of the test samples were low in toxicity at the same concentration. The crude methanolic extract of Zizyphus jujuba showed significant antitermite activity against Heterotermes indicola, among the test samples. Against Tribolium castaneum, Rhizopertha dominica and Callosbruchus analis the insecticidal activity was determined. All the test samples except rc-hexane showed low activity [20%] against T. castaneum. The w-hexane fraction showed low activity [20%] against R. dominica while the rest of the fractions were inactive against it. Low activity of 40% and 20% was shown by the chloroform and "-hexane fraction respectively against C. analis. The results of the present study revealed that the plant could be as potent source of cytotoxic drugs

[ in vitro antifungal effects of Aran extract [Hypericum Perforatum]

The aim of present study was to investigate the antifungal activity of Hypericum perforatum extract in vitro. Three extraction techniques were used [chloroform, acetone and methanol solvents] and the extracts were tested against Fusarium chlamydosporum, Aspergillus niger, Aspergillus fumigatm, Aspergillus flavus. Acetone extract of H. perforatum was the most active against the growth of F. chlamydosporum, A. niger, it caused 55% and 40% inhibition of fungus growth, respectively, when used in a concentration of 7.5 mg/ml from growth media. The fungus A. flavus was more sensitive to the chloroform extract of H. perforatum and the inhibition percentage was 53.84% at the extract concentration of 7.5 mg/ml from growth media. The concentration 7.5 mg/ml of culture media for all extracts was more effective than the concentration 2.5 mg/ml from growth media. It was concluded that the acetone extract of H. perforatum showed a broad spectrum and greatest activity against the fungi among extracts tested

Inhibition of Aspergillus flavus colonization and aflatoxin (AfB1) in peanut by methyleugenol.

Methyleugenol is naturally occurring substance in oils and fruits and in various foods as flavoring agent. Effect of this methyleugenol in inhibiting A. flavus colonization and aflatoxin production on peanut pods and kernels has been studied. Spray of methyleugenol (0.5%) on peanut pods and kernels checked the colonization of A. flavus and aflatoxin synthesis. This chemical can be used as both prophylactic or post infection spray on peanut pods before storage. It is the first report on the inhibition of A. flavus by methyleugenol on peanut.

[Effects of zataria multiflora boiss. essential oil on aspergillus flavus]

Molds are important spoiler of food material and have potential of mycotoxin production. The adverse side effects of chemical preservatives have caused that research focused on use of naturally occurring compound, especially essential oils [EOs], for inhibition of mold growth. Essential oils are natural compounds containing terpenic mixtures that have antimicrobial and antioxidant properties. Zataria multifloro Boiss., a domestic aromatic plant of Iran, has different effect such as antispasmodic and antibacterial. In this study effect of Z. multiflora Boiss. Essential oil on growth and spore production by Aspergillus flavus and its morphological alteration action were investigated. After extraction of essential oil with steam distillation method, it was analyzed by GC-MS. Evaluation of essential oil effect on growth and spore production carried out by agar dilution method. For SEM evaluation, mold cultured on PDA was fixed with osmium tetroxide and blocks of agar were coated with gold and observed by scanning electron microscope. Effect of EO on growth and spore production by A. flavus was significant in all concentrations studied. MIC and MFC values were 400 and 1000 ppm, respectively. SEM analysis showed excessive sporulation of mold culture in absence of EO while sporulation of culture containing EO was very sparse. Morphological changes such as irregular surface with many folds and distortion of hyphae were seen in cultures treated with EO. Inhibitory effect of Z. multiflora Boiss. Essential oil demonstrated in this study candidate this EO as a substitution of chemical preservatives

The efficacy of acetone in the sterilisation of ophthalmic instruments.

Acetone has been considered a quick, effective and less expensive chemical sterilising agent and continues to be used by ophthalmic surgeons, at least in developing countries. Its utility however has been questioned recently. This study was designed to assess the efficacy of acetone against Pseudomonas aeruginosa, Bacillus subtilis and Aspergillus flavus present on ophthalmic surgical instruments (forceps, sutures). The instruments were contaminated by immersion in standard suspensions of the organisms and thereafter were either unwashed (group-I), washed and dried (group-II) or only washed (group-III) before immersion in acetone. The exposure to acetone was kept at 3, 10 and 20 minutes in each group. The results showed that acetone could eliminate Pseudomonas (vegetative bacteria) after 10 minutes exposure in unwashed group and 3 minutes exposure in washed groups. It was ineffective against spore bearing bacteria (B. subtilis) and fungus (Aspergillus flavus) even after 20 minutes of exposure.

Inhibitory effect of boric acid on the development and aflatoxin production by aspergillus flavus and aspergillus parasiticus

El ácido bórico fué analizado en las concentraciones de 0,25 porciento; 0,5 porciento y 1,0 porciento, demostrando su eficacia al inhibir el crecimiento de Aspergillus flavus (NRRL 3251 y NRRL 5520) y Aspergillus parasiticus (UNBF A12 y NRRL 2999), como tambien la producción de aflatoxinas. Los resultados sugieren que este aditivo es eficiente en el control de hongos del grupo A. flavus inhibiendo el crecimiento de los mismos en la concentración máxima permitida por la legislación brasileña (0,5 porciento)

Control of zoopathogenic fungi in vitro by polyamine biosynthesis inhibitors.

Effect of inhibitors of polyamine (PA) biosynthesis, alpha-difluoromethylornithine (DFMO), methylglyoxal bis (guanylhydrazone)--MGBG and bis (cyclohexylammonium) sulphate (BCHA) on mycelial growth of three clinically important fungi-Trichophyton mentagrophytes, Microsporum gypseum and Aspergillus flavus was examined in vitro. All inhibitors at concentrations 1 to 50 mM produced greater inhibition of mycelial growth in all fungi tested in a dose-dependent manner. MGBG was the most effective inhibitor, and T. mentagrophytes was the most sensitive fungus to all inhibitors followed by M. gypseum and A. flavus. The results suggested that control of fungal diseases in animals and human beings with specific inhibitors of PA biosynthesis is possible.

Inhibition of aflatoxin production & growth of Aspergillus flavus by eugenol & onion & garlic extracts.

Efficacy of natural non-toxic materials including extracts of onion and garlic as well as eugenol was tested against aflatoxin production by A. flavus in liquid SMKY medium and in maize grains. Maximum inhibition in the mycelial growth occurred with garlic extract (61.94%), whereas inhibition of aflatoxin production was highest (60.44%) due to onion extract. Eugenol was most suitable for inhibiting aflatoxin production (60.35%) on maize grains.

Influence of sodium bisulfite as an antimicrobial additive compound on growth and nitrogen metabolism of Aspergillus flavus link

The effect of sodium bisulfite on growth and nitrogen metabolism of Aspergilus flavus link was investigated. The concentrations of 0.015, 0.030 and 0.045% were found to inhibit growth of the fungus by 22.1, 51 .3 and 73.4%, respectively. Toal nitrogen was inhibited by 33.1, 43.2 and 75.5% respectively. Protein was also decreased, the percentage of decrease was a factor of sodium bisulfite concentration. Nucleic acids [RNA and DNA] and its related enzymes [nuclease and ribonuclease T1] were inhibited at all experimental sodium bisulfite concentrations, except ribonuclease T1 which showed a significant increase at 0.015%. Total free amino acids were increased at all sodium bisulfite concenrations. The percentage of increases were 285. 7, 359.8 and 94.4% at 0.015,0.03 and 0.045% sodium bisulfite, respectively Methionine appeared at 0.03%, while praline, valine, isoleucine, leucine, tyrosine and phenyl alanine disappeared at 0.045%

Inibiçäo do crescimento de fungos do grupo Aspergillus flavus pelo extrato da semente de "grapefruit" / Growth inhibition moulds of the group Aspergillus flavus by grapefruit seed extract

Nove isolados e uma amostra padräo de fungos do grupo Aspergillus flavus foram avaliados pela sensibilidade ao extrato da semente de "grapefruit". Os fungos foram submetidos a crescimento em ágar contendo 33,3//de extrato de coco-da-bahia e níveis do antifúngico que variavam logaritmamente desde 80 até 5120 ppm. Todas as amostras foram sensíveis ao antifúngico em comportamento linear nas doses testadas, porém, o extrato comportou-se como fungiostático, sendo as doses calculadas para inibiçäo de 99//do crescimento muito elevadas. No entanto, algumas concentraçöes foram satisfatórias, como 5000 ppm (0,5//), que causaram reduçäo de 80//do crescimento na maioria dos isolados

Substâncias antifúngicas, inibidoras de Aspergillus flavus e de outras espécies fúngicas, isoladas de Agave sisalana (sisal) / Antifungal substances, inhibitor of Aspergillus flavus and other fungal species, isolated from Agave sisalana (sisal)

Foram isoladas e caracterizadas parcialmente substâncias antifúngicas a partir de folhas de Agave sisalan (sisal). A atividade inibidora do suco das folhas de sisal foi pronunciada contra Cryptococcus neoformans, Crebrothecium ashbyi e Saccharomyces cerevisiae e menos ativa contra Penicillium chrysogenum, Aspergillus flavus (produtor de aflatoxinas) e A. oryzae. Para o controle rotineiro dos métodos de isolamento do princípio ativo do sisal foi preferido o S. cerevisiae, devido à rapidez do crecimento, inocuidade e sensibilidade. Por meio de participaçäo em solventes e precipitaçöes, foi obtida a sisalanina complexa, que por meio de cromatografia em coluna, foi separada em sete glicosídios designados sisalaninas A, B, C, D, E, F e G. A sisalanina B é constituída de hecogenina, glicose, talactose e xilose, e a sisalanina D dos mesmos compostos e mais a ramnose. A primeira é cerca de quatro vezes mais ativa que a segunda e duas vezes e meia mais que a sisalanina complexa. As concentraçöes inibitórias mínimas da sisalina complexa em ppm determinadas contra fungos säo: A. flavus, 100; A, niger, 1.600; A oryzae, 100; A. parasiticus, 100; Candida albicans, 50; Rhodotorula glutinis, 100 e S.cerevisiae, 12