In this study, we employed a combination of genome mining and heteronuclear single quantum coherence (HSQC)-based small molecule accurate recognition technology (SMART) technology to search for fernane-type triterpenoids. Initially, potential endophytic fungi were identified through genome mining. Subsequently, fine fractions containing various fernane-type triterpenoids were selected using HSQC data collection and SMART prediction. These triterpenoids were then obtained through targeted isolation and identification. Finally, their antifungal activity was evaluated. As a result, three fernane-type triterpenoids, including two novel compounds, along with two new sesquiterpenes and four known compounds were isolated from one potential strain, Diaporthe discoidispora. Their structures were elucidated through analysis of high-resolution electrospray ionization mass spectrometry (HR-ESI-MS) and nuclear magnetic resonance (NMR) spectroscopic data. The absolute configurations were determined using single-crystal X-ray diffraction analysis and electron capture detector (ECD) analysis. Compound 3 exhibited moderate antifungal activity against Candida albicans CMCC 98001 and Aspergillus niger.
Triterpenes/isolation & purification* ; Antifungal Agents/isolation & purification* ; Molecular Structure ; Candida albicans/drug effects* ; Ascomycota/genetics* ; Magnetic Resonance Spectroscopy ; Aspergillus niger/drug effects* ; Genome, Fungal ; Microbial Sensitivity Tests

OBJECTIVE: A strain of Aspergillus niger (A. niger), capable of releasing bound phenolic acids from wheat bran, was isolated. This strain was identified by gene sequence identification. The antioxidant and anti-inflammatory capacity of ferulic acid released from wheat bran by this A. niger strain (FA-WB) were evaluated. METHODS: Molecular identification techniques based on PCR analysis of specific genomic sequences were conducted; antioxidant ability was examined using oxygen radical absorbance capacity (ORAC), cellular antioxidant activity (CAA) assays, and erythrocyte hemolysis assays. RAW264.7 cells were used as a model to detect anti-inflammatory activity. RESULTS: The filamentous fungal isolate was identified to be A. niger. ORAC and CAA assay showed that FA-WB had better antioxidant activity than that of the ferulic acid standard. The erythrocyte hemolysis assay results suggested that FA-WB could attenuate AAPH-induced oxidative stress through inhibition of reactive oxy gen species (ROS) generation. FA-WB could significantly restore the AAPH-induced increase in intracellular antioxidant enzyme activities to normal levels as well as inhibit the intracellular malondialdehyde formation. TNF-a, IL-6, and NO levels indicated that FA-WB can inhibit the inflammation induced by lipopolysaccharide (LPS). CONCLUSION Ferulic acid released from wheat bran by a new strain of A. niger had good anti-inflammatory activity and better antioxidant ability than standard ferulic acid.
Animals ; Anti-Inflammatory Agents ; metabolism ; pharmacology ; Antioxidants ; metabolism ; pharmacology ; Aspergillus niger ; genetics ; isolation & purification ; metabolism ; Coumaric Acids ; metabolism ; pharmacology ; DNA, Fungal ; analysis ; Dietary Fiber ; microbiology ; Erythrocytes ; drug effects ; metabolism ; Fermentation ; Hep G2 Cells ; Humans ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; pharmacology ; Mice ; RAW 264.7 Cells ; Sheep ; Tumor Necrosis Factor-alpha ; metabolism

Cockroaches are abundant in Nigeria and are seen to harbour an array of pathogens. Environmental and sanitary conditions associated with demographic/socio-economic settings of an area could contribute to the prevalence of disease pathogens in cockroaches. A total of 246 cockroaches (Periplaneta americana) in urban (Benin, n=91), semi-urban (Ekpoma, n=75) and rural (Emuhi, n=70) settings in Edo State, Nigeria were collected within and around households. The external body surfaces and alimentary canal of these cockroaches were screened for bacterial, fungal, and parasitological infections. Bacillus sp. and Escherichia coli were the most common bacteria in cockroaches. However, Enterococcus faecalis could not be isolated in cockroaches trapped from Ekpoma and Emuhi. Aspergillus niger was the most prevalent fungus in Benin and Ekpoma, while Mucor sp. was predominant in Emuhi. Parasitological investigations revealed the preponderance of Ascaris lumbricoides in Benin and Emuhi, while Trichuris trichura was the most predominant in Ekpoma. The prevalence and burden of infection in cockroaches is likely to be a reflection of the sanitary conditions of these areas. Also, cockroaches in these areas making incursions in homes may increase the risk of human infections with these disease agents.
Animals ; Ascaris lumbricoides/isolation & purification ; Aspergillus niger/isolation & purification ; Bacillus/isolation & purification ; Cockroaches/*microbiology/*parasitology ; Escherichia coli/isolation & purification ; Nigeria ; Sanitation ; Trichuris/isolation & purification

OBJECTIVETo evaluate the feasibility of PCR/reverse line blot hybridization (RLB) assay in the detection and identification of clinical pathogens in fungal sinusitis (FS).

METHODSTwenty-six formalin-fixed and paraffin-embedded tissues and 8 fresh tissues of FS were collected from Beijing Tongren Hospital, Capital Medical University from May 2009 to February 2010. Pathological examination, fungal culture and ITS2 region sequencing were carried out. The DNA of all samples was extracted by standard procedure and fungal universal primers ITS3 and ITS4 were used for PCR amplification of all tissues. Then the amplified products were used for RLB with five fungal species-specific probes. The results of PCR/RLB were compared with ITS region sequencing, fungal culture and pathological examination.

RESULTSFor the biopsy tissues, fungal cultures were positive in 14 cases (41.2%); pathologic examination demonstrated fungal hyphae in all cases; ITS2 region sequencing was successful in 16 cases (47.1%); PCR/RLB showed A. flavus in 14 cases, A. fumigatus in 10 cases, A. niger in four cases, A. nidulans in one case, A. flavus and A. fumigatus in three cases, and A. fumigatus and A. niger in two cases.

CONCLUSIONSThe PCR/RLB assay is suitable for rapid and accurate detection and identification of the pathogenic fungus of FS. Compared with the conventional fungal culture and microscopy, pathologic examination and DNA sequencing, the PCR/RLB has the advantages of more economy, time saving, and higher sensitivity, specificity and throughput.

Aspergillus ; classification ; genetics ; isolation & purification ; Aspergillus flavus ; genetics ; isolation & purification ; Aspergillus fumigatus ; genetics ; isolation & purification ; Aspergillus niger ; genetics ; isolation & purification ; DNA Primers ; DNA, Fungal ; genetics ; Humans ; Mycoses ; diagnosis ; microbiology ; Nucleic Acid Hybridization ; methods ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Sinusitis ; diagnosis ; microbiology

Streptomyces S24 has broad spectrum against Aspergillus spp. in food and feed, such as Aspergillus flavus, Aspergillus niger and Asperegillus alutacells. The objective of this study was to improve the production of antifungal substances produced by S24 and to test their inhibitory effects on Aspergillus flavus. By using one-factor-at-a-time experiment and orthogonal design method, we optimized the fermentation medium. The composition of an optimized medium for the production of antifungal substances contained (g/L): starch soluble, 10; Glucose, 40; yeast extract, 8; soybean powder, 24; KH2PO4 4; and CaCO3 0.8. As a result, the productivity of antifungal substances could reach to 10 235.45 microg/mL, and this value was 2.81 times higher than that of initial medium before optimization. Additionally, inhibitory effects of the products on Aspergillus flavus were analyzed. Antagonistic tests indicated that the antifungal substances greatly inhibited mycelium growth and spores germination of Aspergillus flavus. We observed through microscope that the mycelia grew abnormally, such as contorting, bulging, vacuole increasing and the cytoplasmic contents inside effusing and the spores appeared unusual, such as gathering, deforming, cytoplasmic contents inside effusing and fracturing.
Adsorption ; Antifungal Agents ; chemistry ; isolation & purification ; pharmacology ; Aspergillus flavus ; drug effects ; growth & development ; Aspergillus niger ; drug effects ; growth & development ; Culture Media ; chemistry ; Food Contamination ; prevention & control ; Streptomyces ; chemistry ; growth & development ; metabolism

Streptomyces S24 has broad spectrum resistance to the Aspergillus in food and feed, such as Aspergillus flavus, Aspergillus niger, Asperegillus alutacells and so on. We studied the adsorption and desorption properties of antifungal substance from Streptomyces S24 on macroporous resins, screened the best elution solution and also investigated some physical and chemical characters of antifungal substance by determining the antifugal activity using oxford plate assay system. According to the analysis results, AB-8 resin offered the best adsorption and desorption capacity for antifungal substance and its saturated absorption capacity was 7.0822 x 10(4) microg/g, the optimal elution solution was 85% acetone and the dynamic desorption rate could reach 93.82%. The antifungal substance was stable to heat and alkali, not sensitive to organic solvents, and sensitive to ultraviolet rays and acid. Based on its ultraviolet spectrometry, the antifungal substance was identified as heptaene macrolide antibiotic.
Adsorption ; Antifungal Agents ; chemistry ; isolation & purification ; pharmacology ; Aspergillus ; drug effects ; Aspergillus flavus ; drug effects ; Aspergillus niger ; drug effects ; Streptomyces ; chemistry ; metabolism

A lipase from Aspergillus niger F044 was purified to homogeneity using ammonium sulfate precipitation, dialysis, DEAE-Sepharose Fast Flow anion exchange chromatography and Sephadex G-75 gel filtration chromatography. This purification protocol resulted in a 73.71-fold purification of lipase with 33.99% final yield, and the relative molecular weight of the enzyme was determined to be approximately 35-40kD using SDS-PAGE. The optimum pH and temperature for lipolytic activity of the lipase was 7.0 and 45 degrees C , respectively. It was extremely stable at 60 degrees C and retained 98.70% of its original activity for 30min. The stability declined rapidly as soon as the temperature rose over 65 degrees C . The lipase was highly stable in the pH range from 2.0 to 9.0 for 4h. Ca2+ and Mg2+ ions stimulated lipolytic activity, whereas Mn2+ , Fe2+ and Zn2+ ions caused inhibition. The values of Km and Vmax calculated from the Lineweaver-Burk plot using pNPP as hydrolysis substrate were 7.37mmol/L and 25.91 micromol/(min x mg), respectively. The N-terminal sequence of the lipase was Ser/Glu/His-Val-Ser-Thr-Ser-Thr-Leu-Asp-Glu-Leu-Gln-Leu-Phe-Ala-Gln, which is highly homogeneity with that of lipase, as reported by Torossian.
Amino Acid Sequence ; Aspergillus niger ; enzymology ; Chromatography, Gel ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Enzyme Stability ; Fungal Proteins ; chemistry ; isolation & purification ; metabolism ; Hydrogen-Ion Concentration ; Lipase ; chemistry ; isolation & purification ; metabolism ; Molecular Sequence Data ; Molecular Weight ; Sequence Analysis, Protein ; Temperature

OBJECTIVETo investigate the contents of glycyrrhizic acid in hejian decoction (mixed the traditional Chinese herbs together, then boiling them with water) and the fenjian decoction (boiling the single traditional Chinese herb with water separately, then mixed the abstracts) of Sanaotang (composed of Ephedra sinica, Prunus armeniaca and Glycyrrhiza uralensis) and to compare with their anti-bacterial activities in vitro.

METHODA HPLC method was established with a Agilent Zorbax SB-C18 column (4. 6 mm x 250 mm, 5 microm), a mobile phase of acetonitrile-0.2% acetic acid solution (35:65), a flow rate of 1 mL x min(-1) and a detection wavelength of 254 nmn in order to determine the contents of glycyrrhizic acid minimal bacterial inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) antagonized the common bacteria in different decoctions were rieasured in vitro by employing dilution method.

RESULTThe average content of glycyrrhizic acid of the hejian decoction was higher than that of the fenjian decoction. The hejian decoction could display the inhibitory bactericidal activity to Aeruginosus bacillus, but the fenjian decoction could not. And to Staphylococcus aureus, the inhibitory bactericidal activity the hejian decoction was slightly stronger than that of the fenjian decoction.

CONCLUSIONComparing with that of the fenjian decoction, the content of glycyrrhizic acid of the hejian decoction was higher and the anti-bacterial activities was stronger.

Anti-Bacterial Agents ; analysis ; pharmacology ; Antifungal Agents ; analysis ; pharmacology ; Aspergillus niger ; drug effects ; growth & development ; Bacteria ; drug effects ; growth & development ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ephedra sinica ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Glycyrrhizic Acid ; analysis ; pharmacology ; Hot Temperature ; Microbial Sensitivity Tests ; Plants, Medicinal ; chemistry ; Prunus ; chemistry ; Pseudomonas aeruginosa ; drug effects ; growth & development ; Staphylococcus aureus ; drug effects ; growth & development

OBJECTIVETo establish the validation method and criteria for counting bacteria and fungi in microbial limit test which is described in the Pharmacopeia of China (ChP) 2005.

METHODAccording to the method set up for validation, the tested microorganisms with known counts were added to samples followed by the determination of the recovery.

RESULTWith different preparing method for testing samples, the recoveries for the tested microorganisms in testing samples were found to be over 70%.

CONCLUSIONValidation method for counting contaminated bacteria and fungi in drugs is recommended to follow the method established in this paper. The recovery for tested microorganisms should be not less than 70%.

Aspergillus niger ; isolation & purification ; Bacillus subtilis ; isolation & purification ; Bacteria ; isolation & purification ; Candida albicans ; isolation & purification ; Colony Count, Microbial ; methods ; Drug Contamination ; Drugs, Chinese Herbal ; standards ; Escherichia coli ; isolation & purification ; Fungi ; isolation & purification ; Staphylococcus aureus ; isolation & purification

OBJECTIVETo investigate the removal of cadmium from aqueous solution by waste fungal biomass of Aspergillus niger, originated from citric acid fermentation industry.

METHODSBatch adsorption test was used to study the biosorption equilibrium and isotherm. The Cd2+ concentration was measured with atomic adsorption spectrophotometer (AAS) HITACHI 180-80.

RESULTSThe biosorption achieved equilibrium within 30 min. The adsorption isotherm could be described by Freundlich adsorption model, and the constants K(F) and 1/n were determined to be 2.07 and 0.18, respectively, and the correlation efficiency was 0.97. The optimal pH for Cd adsorption was 6.0. The cadmium-laden biomass could be effectively regenerated using 0.1 N HCl.

CONCLUSIONThe waste biomass of Aspergillus niger, a by-product of fermentation industry, is a potential biosorbent for the removal of cadmium from aqueous solution.

Adsorption ; Aspergillus niger ; Biomass ; Cadmium ; isolation & purification ; Citric Acid ; Hydrogen-Ion Concentration ; Industrial Waste ; Waste Disposal, Fluid ; Water Pollutants, Chemical ; isolation & purification ; Water Purification ; methods