Fungi are frequently infected with viruses called mycoviruses. Some mycoviruses have shown potential as biocontrol agents because they can weaken the virulence of fungal plant pathogens. Intensive research has been conducted on the diversity of viruses infecting plant pathogens and the functions of such viruses. In contrast, the diversity of viruses infecting fungi that are not plant pathogens is less well understood. We previously identified a mycovirus of the family Partitiviridae, Aspergillus sulphureus partitivirus 1 (AsuPV1), from an Aspergillus sulphureus strain isolated from fermented dried bonito. In the present study, we conducted a detailed phylogenetic analysis of AsuPV1 and determined its molecular characteristics. The phylogenetic analysis revealed that AsuPV1 belongs to the proposed subgroup I within the genus Gammapartitivirus. Virus particles of AsuPV1 were isolated by density gradient centrifugation, and molecular analyses indicated that each of its three genome segments is encapsulated in capsids composed of the putative viral coat protein. AsuPV1 is the second virus with a tri-segmented genome to be found in a member of Gammapartitivirus subgroup I, after Penicillium stoloniferum virus F. A phenotypic comparison between the parental A. sulphureus strain and isogenic virus-free isolates indicated that AsuPV1 enhances conidium production in its host. This study provides insight into the diversity and molecular characteristics of partitiviruses with a tri-segmented genome.

Trichoderma asperellam is a fungal species that is frequently utilized in the biological control of plant pathogenic fungi.The creation of analytical enzymes enhances its ability to control biological infections.It has been highlighted that physical traits alone are insufficient for identifying these fungal species.As a result, the study's goal was to use molecular markers to identify Trichoderma species and assess their antagonistic effectiveness against plant pathogens.T. asperellam have antagonistic and physiologically controlled activity, which inhibited the pathogenic fungus to the greatest extent possible in a dual culture approach.T. asperellam had the highest inhibition (62.7%), followed by T. harzianum (56.0%), T. koeningii (56.0%),Aspergillus niger (52.5%),Aspergillus sulphureus (48.5%),Penicillium sp.(48.41%), and Aspergillus flavus (47.3%).

The physicochemical and bacteriological quality of borehole, bottle and sachet water sold in Anchor University and within Ayobo community Lagos state, Nigeria was investigated Microbiological analysis was carried out using standard microbial procedure to ensure that the water is microbiologically safe. It was screened for the presence of coliforms and other pathogenic microorganisms. The total heterotrophic bacterial count for bottle, sachet and borehole water are 16.50-123.50 x 103CFU/ml, 65.00-73.00 x 103CFU/ml and 0.00-72.00 x 103CFU/ml respectively while the total heterotrophic fungal count for bottle, sachet and borehole water are 5.00-54.50 x 103CFU/ml, 11.00-27.50 x 103CFU/ml and 6.00-16.16.00 x 103CFU/ml respectively. Most probable number was determined using membrane filtration method and it ranged from 15MPN/100ml, 22MPN/100ml, and 27MPN/100 ml for bottle, sachet and borehole water respectively. The mean total coliform per 100ml ranged from 22-30 x 103CFU/ml while fecal coliform ranged from 4-11 x 103CFU/ml. The isolated organisms were Salmonella paratyphi, Shigella flexneri, Proteus vulgaris, Pseudomonas aeroginosa, Escherichia coli, Citrobacter freudii, Salmonella paratyphi, Aspergillus candidus, Aspergillus niger, Aspergillus sulphureus and Penicillium corylophilum. Most of the bacteria isolated showed multidrug resistance to Augmentin, Gentamycin, Pefloxacin, Tarivid, Streptomycin, Septrin, Chloramphenicol and Amoxacillin and showed susceptibility to Ciprofloxacin. The study therefore concludes that these water samples do not meet the WHO standards for potable water; hence they can be potential sources of waterborne diseases.

Characterization of a novel GH10 xylanase with a carbohydrate binding module from Aspergillus sulphureus and its synergistic hydrolysis activity with cellulase

Research into radionuclide leaching behavior in the treatment of low-level radioactive waste solidification has been a widespread concern. Bioleaching of uranium was carried out using Aspergillus sulphureus isolated from El-Sella ore material. This area is located in the southern eastern desert of Egypt. The maximum bioleaching of U(VI) was found to be 83.1 % at these optimum conditions: 9 days incubation period, 0.5% pulp density, 30 °C incubation temperature at shaking speed 175 rpm. Glucose and NH4Cl were considered the best carbon and nitrogen sources for fungal growth through direct bioleaching process. A. sulphureus exhibits good potential in generating varieties of organic acids which effective for bioleaching of uranium. It produced 45.2 and 17 µg/ml from gallic and ellagic acids, respectively in the culture filtrate under the optimum conditions. This work represented the environmental technology for leaching of uranium where this is method is more save and low cost.

In Cameroon, palm oil is extensively used in its crude form for food. The present study was carried out to access stakeholders’ perception of spoilage and the microbial quality of crude palm oil in the North-west Region of Cameroon.  A random survey was carried out on 148 stakeholders of the crude palm oil marketing channel about their perception of crude palm oil spoilage. 79 samples were collected from different market sites in the region. Handlings of crude palm oil by the stakeholders were unhygienic and they had poor knowledge of the causes and reasons associated with its spoilage. The microbial contaminants isolated were Salmonella sp., Escherichia coli, Staphylococcus aureus, yeast, and Aspergillus niger, Aspergillus sulphureus and Aspergillus versicolor. The estimated overall bacterial load ranged from 17.14 × 104 to 36.41 ×104cfu/ml. The bacterial load of crude palm oil samples from each market was far above the minimum acceptable range stipulated by NAFDAC. There is need for these stakeholders to be educated on the health implications and risks associated with palm oil production and post-production handling.    Key words: Crude palm oil, spoilage, microbiological quality, Northwest Region of Cameroon.

This study was done on a piece of archaeological leather document preserved in Egyptian museum storeroom under No.JE90809 SR 5/13677, excavated 1966. The leather document showed different aspects of deterioration types. Different analytical methods were used for identifying the components of the leather document to explain its mechanism of deterioration process. Analytical methods were used: visual assessment, documentation process by high resolution camera and scanner in multispectral imaging (MSI), diagnostic examinations using Multi Spectral Imaging (VIS-UV-IR), isolation and identification of different types of fungi, investigation of the surface morphology and animal type using digital light microscope (Dino light), stereo microscope (S.M.) and polarizing microscope (P.M.), identification of ink binder, amino acids degradation and tanning materials by Fourier transformation infrared spectrophotometry (FTIR) it is a qualitative analysis to give general information concerning collagen, identification of inks by X-ray diffraction (XRD), measuring the thickness by micro meter. In addition to suggest a new method for Museum show for the archaeological leather document as the preparation of a new method of exhibition is considered one of the most important goal of this study, because the current method of exhibition is considered one of the most important reason for the deterioration of the studied document. The results revealed thatthe microscopic examinations of samples clarify that the type of skin used was sheep in comparison with the standard samples. Also it explained the deformation of the appearance, contaminations from stains and dusts, and damages caused by physical factors. X-ray diffraction results showed that the ink used was made of carbon black ink. In addition the results showed that thickness measurement numbers were different which give indicate to somehow the manufacturing process was not so perfect. Also the results of FTIR proved the degradation of the collagen in the archaeological leather document, the binding agent was Arabic gum and the leather was tanned with vegetable tanning. Isolation and identification of micro-organisms clarified the most dominant fungi isolated from the archaeological leather document were: Aspergillus niger, Aspergillus sulphureus, Aspergillus versicolor, Aspergillus sydowii, Penicillium chrysogenum, Penicillium islandicum, Alternaria alternate, Aspergillus flavus,Aspergillus terreus. Finally, the storage of the leather document was very poor which led to different aspects of deterioration. That prompted the author to suggest a new method for exhibition for the archaeological leather document. ملخص البحث باللغة العربية قطعة من الجلد الأثري محفوظة في مخزن المتحف المصري برقم JE90809 5/13677 ، تم اکتشافها عام 1966 ، وتعانى الوثيقة الجلدية مظاهر التلف المختلفة. ترکز هذه الدراسة على استخدام التقنيات التحليلية الحديثة لتحديد مکونات المخطوطة لشرح عمليات التلف. استخدمت فى الدراسة التقييم البصري ، وعملية التوثيق بواسطة کاميرا عالية الدقة والماسح الضوئي في التصوير متعدد الأطياف (MSI) ، والفحوص التشخيصية باستخدام التصوير متعدد الأطياف (VIS-UV-IR) ، وعزل وتحديد الفطريات ، والتحقيق في الشکل السطحي و نوع الحيوان باستخدام أنواع مختلفة من المجاهر: المجهر الضوئي الرقمي، المجهر الاستريو (SM) ، المجهر الاستقطابي (PM) ، ايضا تم تحديد نوع الحبر ، ومواد الدباغة بواسطة التحليل الطيفي بالأشعة تحت الحمراء (FTIR) ، وتم تحديدنوع الحبر بواسطة حيود الأشعة السينية (XRD) ، وتم قياس السمک للقطعة. بالإضافة إلى اقتراح طريقة جديدة للعرض المتحفى للوثيقة حيث أن طريقة العرض الحالية هي إلى حد ما السبب الذي تسبب في تلف القطعة. وقد أوضحت نتائج الفحوصات للعينات أن نوع الجلد المستخدم هو الأغنام مقارنة بالعينات القياسية. کما أظهرت نتائج حيود الأشعة السينية أن الحبر الأسود المستخدم هو الحبر الکربونى. بالإضافة إلى ذلک ، أظهرت النتائج اختلافا في أرقام نتائج قياس السمک للقطعة مما يدل بطريقة أو بأخرى على أن عملية التصنيع لم تکن مثالية. کما أثبتت نتائج FTIR تلف وتدهور الکولاجين في الوثيقة الجلدية ، وکان الوسيط الصمغ العربي والجلد المدبوغ تم دباغته بالتانين (الدباغة النباتية). أوضح عزل الکائنات الحية الدقيقة أن الفطريات المعزولة من هي: Aspergillus niger و Aspergillus sulphureus و Aspergillus versicolor و Aspergillus sydowii و Penicillium chrysogenum و Penicillium islandicum و Alternaria Alternus و Aspergillgillus flavus. وأخيرا ، کان تخزين المخطوطة رديئا للغاية مما أدى إلى جوانب مختلفة من التدهور. ودفع ذلک المؤلفين إلى اقتراح طريقة جديدة للعرض المتحفى للوثيقة الجلدية الأثرية.

Bed linen is clearly recognized as a potential reservoir for microorganisms and could be a vector of disease transmission. The present study was aimed at isolating and characterizing bacteria and fungi from different kinds of bed linen of student in Ekiti State University hostels. Pour plate method was used for the enumeration of total bacteria count from the posterior and anterior end of the bed linen. The average bacteria count for the anterior and posterior end was 7.46 and 7.16 Log10 CFU/ml respectively. The most dominant microbial species were bacteria and these were mostly found in the environment and on human skin. The bacteria isolated were Staphylococcus aureus, Staphylococcus epidermidis, Proteus mirabilis, Enterobacter aerogenes, Bacillus cereus, Escherichia coli and Klebsiella aerogenes. Bacillus cereus had the highest frequency of occurrence (25%), followed by Staphylococcus aureus (15%), Escherichia coli (15%), Klebsiella aerogenes (15%), Staphylococcus epidermis (15%), Enterobacter aerogenes (10%) and Proteus mirabilis (5%). The fungi isolates were Aspergillus sulphureus, Aspergillus fumigatus, Aspergillus niger, Aspergillus flavus and Fusarium spp. Antibiotics susceptibility test was carried out on the bacteria isolates with gram negative bacteria showing resistance to Cotrimoxazole and gram positive bacteria showing resistance to Amplicillin. Most of the bacteria isolates have multiple antibiotics resistance. The bacterial isolates were susceptible to Ciprofloxacin. Plasmid profiling was also done with Escherichia coli having three high molecular weight plasmids, Bacillus cereus and Enterobacter aerogenes do not possess plasmid. The identified species are suspected to be opportunistic pathogens for human, representing a risk for people with weakened immune system, especially in cases of super-infection.

Wheat and chickpea are considered the most important sources of food and energy and stored for longer period of time in the rural areas in the farmers sheds as seed for the next crop and as market commodity to sale. Both of these crops are liable to many mycoflora which not only deteriorate their quality but also leads toward the crop failure during the coming season. A number of seed borne pathogens viz., Aspergills niger, Aspergillus flavus, Aspergillus sulphureus, Aspergillus nidulense, Rhizopus stolonifer, Penicillium spp. and Drechslera spp, were isolated from wheat seed samples while Aspergillus niger, Aspergillus flavus, Bipolaris spp., Penicillium spp. and Rhizopus stolonifer were isolated from seed samples of chickpea. Various broad spectrum fungicides were evaluated against the isolated mycoflora which showed significant result. Score, Topsin-M and ridomil gold to be the most significant fungicides at all doses against the isolated mycoflora.

Thirty polymorphic simple sequence repeat (SSR) markers have been developed and characterized from RAPD product in Aspergillus niger in order to protect its natural resources. Polymorphisms of these RAPD-SSR markers were evaluated in a natural population of 25 strains collected from Shandong in China. The number of alleles (Na) per locus varied from 2 to 9. Observed (HO) and expected (HE) heterozygosities ranged from 0.56 to 0.96, and from 0.52 to 0.88, respectively. All the RAPD-SSR loci conformed to Hardy–Weinberg equilibrium after Bonferroni correction. They have showed sufficient level of polymorphisms to estimate the genetic diversity, population structure and species conservation in the A. niger. Five additional fungus species, Penicillium chrysogenum, Penicillium citrinum, Aspergillus flavus, Aspergillus ochratoxin and Aspergillus sulphureus were assessed for cross-species amplification. Three of the five species showed at least three polymorphic loci. In addition, eight loci were found to be polymorphic in at least one species.

Aspergillus fresenii and Aspergillus sulphureus produce ochratoxin A (OTA), which is a secondary metabolite of Aspergillus and Penicillium species, with nephrotoxic effects and potential carcinogenic activity. The aim of this study was to determine the effects of temperature (20, 30, and 37°C), water activity (0.82, 0.86, 0.90, 0.94, and 0.98aw), incubation period (5, 10, 15, and 30days) on fungal growth, and OTA production by A. fresenii and A. sulphureus on Nyjer oil seeds. There was no fungal growth at 0.82aw. The two fungal species were able to produce OTA from the fifth day of incubation from 0.86 to 0.98aw and temperature 20 to 37°C. Aspergillus fresenii produced the highest concentration of OTA (643μg/kg) at 0.90aw and 37°C within 15days, while A. sulphureus produced the highest level of OTA (724μg/kg) at 0.98aw and 20°C within 10days. The optimum water activity and temperature for the growth of both fungi were similar at 0.94aw and at 30°C. There was statistically significant difference between the levels of OTA production among the two fungi. Overall, A. sulphureus produced significantly higher levels of OTA (p < 0.05). Higher temperature (37°C) and 0.90-0.94aw were optimal for OTA production by A. fresenii. Our results show that Nyjer seeds can support the growth of A. fresenii and A. sulphureus and OTA production, and the two species had similar temperature and water activity requirements for growth but different requirements for OTA production on Nyjer seeds.

Five new prenylated indole alkaloids, 17-hydroxynotoamide D (1), 17-O-ethylnotoamide M (2), 10-O-acetylsclerotiamide (3), 10-O-ethylsclerotiamide (4), and 10-O-ethylnotoamide R (5) were isolated from a co-culture of marine-derived fungi Aspergillus sulphureus KMM 4640 and Isaria felina KMM 4639. The structures of 1-5 were determined by detailed analysis of spectroscopic data and by comparison with related known compounds. The absolute configurations of 1-5 were determined by time-dependent density functional theory (TD-DFT) calculations of ECD spectra. Compound 2 is able to inhibit the colony formation of human prostate cancer cells 22Rv1 at non-cytotoxic concentration of 10 μM.

ObjectiveThis study was designed to investigate the effects of an Aspergillus sulphureus xylanase expressed in Pichia pastoris on the growth performance, nutrient digestibility and gut microbes in weanling pigs.MethodsA total of 180 weanling pigs (initial body weights were 8.47±1.40 kg) were assigned randomly to 5 dietary treatments. Each treatment had 6 replicates with 6 pigs per replicate. The experimental diets were wheat based with supplementation of 0, 500, 1,000, 2,000, and 4,000 U xylanase/kg. The experiment lasted 28 days (early phase, d 0 to 14; late phase, d 15 to 28).ResultsIn the early phase, compared to the control, average daily gain (ADG) was higher for pigs fed diets supplemented with xylanase and there was a quadratic response in ADG (p<0.05). In the entire phase, ADG was higher for the pigs fed 1,000 or 2,000 U/kg xylanase compared to the control (p<0.05). The gain to feed ratio was higher for pigs fed diets supplemented with 1,000 or 2,000 U/kg xylanase compared to the control (p<0.05). Increasing the amount of xylanase improved the apparent total tract digestibility of dry matter, crude protein, neutral detergent fiber, calcium, and phosphorus during both periods (p<0.05). Xylanase supplementation (2,000 U/kg) decreased the proportion of Lachnospiraceae (by 50%) in Firmicutes, but increased Prevotellaceae (by 175%) in Bacteroidetes and almost diminished Enterobacteriaceae (Escherichia-Shigella) in Proteobacteria.ConclusionXylanase supplementation increased growth performance and nutrient digestibility up to 2,000 U/kg. Supplementation of xylanase (2,000 U/kg) decreased the richness of gut bacteria but diminished the growth of harmful pathogenic bacteria, such as Escherichia-Shigella, in the colon.

As a feed additive, xylanase has been widely applied in the feed of monogastric animals, which contains multiple plant polysaccharides. However, during feed manufacture, the high pelleting temperatures challenge wild-type xylanases. The aim of this study was to improve the thermostability of Aspergillus sulphureus acidic xylanase. According to the predicted protein structure, a series of disulphide bridges and proline substitutions were created in the xylanase by PCR, and the mutants were expressed in Pichia pastoris. Enzyme properties were evaluated following chromatographic purification. All the recombinant enzymes showed optima at pH 3.0 and 50 °C or 55 °C and better resistance to some chemicals except for CuSO4. The specific activity of the xylanase was decreased by introduction of the mutations. Compared to the wild-type enzyme, a combined mutant, T53C-T142C/T46P, with a disulphide bond at 53–142 and a proline substitution at 46, showed a 22-fold increase of half-life at 60 °C. In a 10-L fermentor, the maximal xylanase activity of T53C-T142C/T46P reached 1,684 U/mL. It was suggested that the T53C-T142C/T46P mutant xylanase had excellent thermostability characteristics and could be a prospective additive in feed manufacture.

Context: Sida acuta Burm.f. (Malvaceae) extracts are reported to have applications against malaria, diuretic, antipyretic, nervous and urinary diseases. No fungal endophytes of S. acuta are reported. Objective: Isolation, identification and evaluation of antibacterial, antioxidant, anticancer and haemolytic potential of fungal endophytes from the ethnomedcinal plant S. acuta. Materials and methods: Sida acuta stem segments were placed on PDA medium to isolate endophytic fungi. The fungus was identified by genomic DNA analysis and phylogenetic tree was constructed using ITS sequences (GenBank) to confirm species. The antibacterial efficacy of Aspergillus sulphureus MME12 ethyl acetate extract was tested against Gram-positive and Gram-negative pathogenic bacteria. DPPH free radical scavenging activity, anticancer and DNA fragmentation against EAC cells, and direct haemolytic activity (100–500 μg/mL) using human erythrocytes were determined. Results and discussion: The ethyl acetate extract of A. sulphureus (Fresen.) Wehmer (Trichocomaceae) demonstrated significant antibacterial potential against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Salmonella typhi compared to streptomycin. MIC against test pathogens was in the range of 15.6–62.5 μg/mL. The antioxidant results revealed significant RSA from 12.43% to 62.02% (IC50 = 350.4 μg/mL, p ≤ 0.05). MME12 offered considerable inhibition of EAC proliferation (23% to 84%, IC50 = 216.7 μg/mL, p ≤ 0.05) supported by DNA fragmentation studies. The extract also offered insignificant haemolysis (5.6%) compared to Triton X-100. Conclusions: A single endophytic fungus, A. sulphureus MME12 was isolated and identified using molecular profiling. The above-mentioned findings support the pharmacological application of A. sulphureus MME12 extract and demand for purification of the active principle(s).

New diorcinol J (1) and four known diorcinols B-E (2-5) were isolated from the EtOAc extract of a co-culture of marine isolates of the fungi Aspergillus sulphureus KMM 4640 and Isaria felina KMM 4639. The structure of diorcinol J was established using NMR spectroscopy and high-resolution mass spectrometry. The cytotoxicity of the isolated compounds and their ability to enhance expression of heat shock protein Hsp70 in Ehrlich ascites carcinoma cells were investigated.

Growth regulating activity of metabolites of terrestrial plants and marine organisms – water, alcohol and lipidic extracts, polysaccharides, diterpene, triterpene and steroidal glycosides, alkaloids, pigments, decumbenones and B,B'-triketones were studied. It was defined that some of the studied substances are capable to stimulate, inhibit or be inactive in relation to the main root of seedlings of agricultural plants. Extract and triterpene glycoside of hederagenin cauloside C, isolated from Caulophyllum robustum Maxim., and monosulfated cucumariosides group A 2, isolated from of the Far Eastern edible holothurian Cucumaria japonica, inhibit the growth of seedlings the main root of Cucumis sativus. Among of decumbenones A (1), B (2) and C (3), isolated from the marine-derived strain of the fungus Aspergillus sulphureus, decumbenone A (1) stimulates the growth of seedling roots of Triticum aestivum and Zea mays . Among of alkaloids 1-9, isolated from the marine fungus Aspergillus fumigatus, of alkaloid 3 stimulated the growth of seedling roots of ?rysa sativa and Fagopyrum esculentum . The water extracts of F. esculentum (of the red-stem population) at a concentration of 10 -8 g/ml stimulated productivity of one plant F. esculentum by 67% and content of rutin in F. esculentum plants by 20% more than in the control.

A new decaline derivative, decumbenone C (1) along with four known compounds, decumbenones A (2) and B (3), diorcinol (4), and brevianamide F (5) were isolated from the marine fungus Aspergillus sulphureus KMM 4640. Decumbenone C shows potent cytotoxic activity against SK-MEL-5 human melanoma cells with IC(50) values of 0.9 μM.

Two chimeric genes, XynA-Bs-Glu-1 and XynA-Bs-Glu-2, encoding Aspergillus sulphureus β-xylanase (XynA, 26 kDa) and Bacillus subtilis β-1,3-1,4-glucanase (Bs-Glu, 30 kDa), were constructed via in-fusion by different linkers and expressed successfully in Pichia pastoris. The fusion protein (50 kDa) exhibited both β-xylanase and β-1,3-1,4-glucanase activities. Compared with parental enzymes, the moiety activities were decreased in fermentation supernatants. Parental XynA and Bs-Glu were superior to corresponding moieties in each fusion enzymes because of lower Kn higher kcat. Despite some variations, common optima were generally 50°C and pH 3.4 for the XynA moiety and parent, and 40°C and pH 6.4 for the Bs-Glu counterparts. Thus, the fusion enzyme XynA-Bs-Glu-1 and XynA-Bs-Glu-2 were bifunctional.

The effect of decumbenones A (1), B (2) and C (3) from the marine-derived strain of the fungus Aspergillus sulphureus on the growth of seedling roots of buckwheat, wheat, barley and corn at the concentration range 10﹣5 - 10﹣18 M was studied. It was shown that decumbenone B had a stimulatory effect on the growth of seedling roots of buckwheat, decumbenone A—on the growth of seedling roots of spring soft wheat, decumbenone C—on the growth of seedling roots of spring barley, decumbenone A, B and C —on the growth of seedling roots of corn. The stimulatory effect for some substances was shown at ultra-low concentrations 10﹣12 - 10﹣18 M. It is possible to recommend decumbenones A, B and C for studying in field conditions as growth factors of buckwheat, wheat, barley and corn.