Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 56

Search results for: Aspergillus flavus

56 Solid-State Bioconversion of Pineapple Residues into Kojic Acid by Aspergillus flavus: A Prospective Study

Authors: S. Nurashikin, E. Z. Rusley, A. Husaini

Abstract:

Kojic acid is an organic acid that is widely used as an ingredient for dermatological products, precursor for flavor enhancer and also as anti-inflammatory drug. The present study was undertaken to test the feasibility of pineapple residues as substrate for kojic acid production by Aspergillus flavus Link 44-1 via solid-state fermentation. The effect of initial moisture content, pH and incubation time on kojic acid fermentation was investigated. The best initial moisture content for kojic acid production from pineapple residues was observed at 70% (v/w) whereas initial culture pH 2.5 was identified to give high production of kojic acid. The optimal range of incubation time was identified between 8 and 14 days of incubation which corresponded to highest range of kojic acid produced. The results from this study pronounce the promising usability of pineapple residues as alternative substrate for kojic acid production by A. flavus Link 44-1.

Keywords: Aspergillus flavus, kojic acid, pineapple residues, solid state fermentation.

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55 Isolation and Screening of Fungi for Aerobic Delignification and Reduction of AOX of Pulp and Paper Mill Effluent

Authors: N. Lokeshwari, G Srinikethan, S. G. Joshi, I. Shasikala, B. Srikanth, Bashirahmed, L. Sushma

Abstract:

Water pollution is a major concern for the pulp and paper industry due to the large quantities of effluents generated. Biodegradation of industrial Lignin and AOX by a fungal isolate identified as Aspergillus flavus, white rot fungi which was isolated from Pulp and Paper effluent was studied in batch flask system with industrial effluent and synthetic solution. The flasks were operated at temperature 32°C at 200rpm for eight days in continuous mode. The average overall pH, Temperature, DO, C.O.D, T.D.S, T.S.S, Lignin, AOX were up to 4.56, 32oC, 4.2mg/l, 104mg/l, 6000 mg/l, 4000mg/l, 575.5mg/l, 2195 mg/l respectively after treatment. The Aspergillus flavus sp was the most effective in the biodegradation of Lignin of pulp industry for 94% at 480nm, AOX for 62% at 510nm and of chemical oxygen demand levels for 45% after 8 days of incubation. The optimal conditions found were 4 pH and 32oC temperature for lignin and AOX degradation.

Keywords: Aspergillus flavus, Lignin, Optimal conditions, Quantification studies.

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54 Control of Aspergillus flavus Growth in Tomato Paste by Cinnamomum zeylanicum and Origanum vulgare L. Essential Oils

Authors: F. Kalantari, M. Barzegar, Z. Hamidi-Esfahani

Abstract:

This study was conducted to evaluate the antifungal activities of Cinnamomum zeylanicum and Origanum vulgare L. essential oil against Aspergillus flavus in culture media and tomato paste. 200 ppm of cinnamon and 500 ppm of oregano completely inhibited A. flavus growth in culture media, while in tomato paste 300 ppm of cinnamon and 200 ppm of oregano had the same effect. Test panel evaluations revealed that samples with 100 and 200 ppm cinnamon were acceptable. The results may suggest the potential use of Cinnamomum zeylanicum essential oil as natural preservative in tomato paste.

Keywords: Antimicrobial, Food safety, GC/MS, Natural preservative

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53 Inhibitory Effect of Helichrysum arenarium Essential Oil on the Growth of Food Contaminated Microorganisms

Authors: Ali Mohamadi Sani

Abstract:

The aim of this study was to determine the antimicrobial effect of Helichrysum arenarium L. essential oil in "in-vitro" condition on the growth of seven microbial species including Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Saccharomyces cereviciae, Candida albicans, Aspergillus flavus and Aspergillus parasiticus using micro-dilution method. The minimum inhibitory concentration (MIC) and minimum bactericidal or fungicidal concentration (MBC, MFC) were determined for the essential oil at ten concentrations. Finally, the sensitivity of tested microbes to essential oil of H. arenarium was investigated. Results showed that Bacillus subtilis (MIC=781.25 and MBC=6250 µg/ml) was more resistance than two other bacterial species. Among the tested yeasts, Saccharomyces cereviciae (MIC=97.65 and MFC=781.25 µg/ml) was more sensitive than Candida albicans while among the fungal species, growth of Aspergillus parasiticus inhibited at lower concentration of oil than the Aspergillus flavus. The extracted essential oil exhibited the same MIC value in the liquid medium against all fungal strains (48.82 µg/ml), while different activity against A. flavus and A. parasiticus was observed in this medium with MFC values of 6250 and 390.625µg/ml, respectively. The results of the present study indicated that Helichrysum arenarium L essential oil had significant (P<0.05) antimicrobial activity; therefore, it can be used as a natural preservation to increase the shelf life of food products.

 

Keywords: Helichrysum arenarium, Antimicrobial agent, Essential oil, MIC.

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52 Effect of Entomopathogenic Fungi on the Food Consumption of Acrididae Species

Authors: S. Kumar, R. Sultana

Abstract:

This study was conducted to evaluate the effect of Aspergillus species on acridid populations which are major agricultural pests of rice, sugarcane, wheat, maize and fodder crops in Pakistan. Three and replicates i.e. Aspergillus flavus, A. fumigatus and A. niger, excluding the control, were held under laboratory conditions. It was observed that consumption faecal production of acridids was significantly reduced after the pathogenic application of Aspergillus. In the control replicate, the mortality ratio for stage (N4-N6) was maximum on day 2nd i.e. [F10.7 = 18.33, P < 0.05] followed by [F4.20 = 07.85, P < 0.05] and [F3.77 = 06.11, P < 0.05] on 4th and 3rd day, respectively. Similarly, it was a minimum i.e. [F0.48 = 84.65, P < 0.05] on the 1st day. It was also noted that faecal production of Acridid nymphs was not significantly affected when treated with conidial concentration in H2O formulation; however, it was significantly reduced after the contamination with conidial concentration in oil. The high morality of acridids after contamination of Aspergillus supports their use as bio-control agent for reducing pest population. The present study recommends that exploration and screening must be conducted to provide additional pathogens for evaluation as potential biological control against grasshoppers and locusts.

Keywords: Acridid, agriculture, Aspergillus, formulation, Grasshoppers.

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51 Antimicrobial Activity and Phytochemicals Screening of Jojoba (Simmondsia chinensis) Root Extracts and Latex

Authors: Ferial M. Abu-Salem, Hayam M. Ibrahim

Abstract:

Plants are rich sources of bioactive compounds. In this study the photochemical screening of hexane, ethanolic and aqueous extracts of roots and latex of jojoba (Simmondsia chinensis) plant revealed the presence of saponins, tannins, alkaloids, steroids and glycosides. Ethanolic extract was found to be richer in these metabolites than hexane, aqueous extracts and latex. The extracts and latex displayed effective antimicrobial activity against Salmonella typhimurium, Bacillus cereus, Clostridium perfringens, Staphylococcus aureus, Escherichia coli, Candida albicans and Aspergillus flavus. The increase in volume of the extracts and latex caused more activity, as shown by zones of inhibition. Candida albicans growth was inhibited only by hexane extract. Jojoba latex was not effective against Candida albicans at 0.1 and 0.5 ml extracts concentration but showed 5mm zone of inhibition at (1.0 ml). Lower volume (0.1ml) of latex encouraged Aspergillus flavus growth, while at (1.00 ml) reduced its mycelial growth. Thus, jojoba root extracts and latex can be of potential natural antimicrobial agents.

Keywords: Antimicrobial activity, Jojoba (Simmondsia chinensis), latex, photochemical, root Extracts.

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50 Kinetic Studies on Microbial Production of Tannase Using Redgram Husk

Authors: S. K. Mohan, T. Viruthagiri, C. Arunkumar

Abstract:

Tannase (tannin acyl hydrolase, E.C.3.1.1.20) is an important hydrolysable enzyme with innumerable applications and industrial potential. In the present study, a kinetic model has been developed for the batch fermentation used for the production of tannase by A.flavus MTCC 3783. Maximum tannase activity of 143.30 U/ml was obtained at 96 hours under optimum operating conditions at 35oC, an initial pH of 5.5 and with an inducer tannic acid concentration of 3% (w/v) for a fermentation period of 120 hours. The biomass concentration reaches a maximum of 6.62 g/l at 96 hours and further there was no increase in biomass concentration till the end of the fermentation. Various unstructured kinetic models were analyzed to simulate the experimental values of microbial growth, tannase activity and substrate concentration. The Logistic model for microbial growth , Luedeking - Piret model for production of tannase and Substrate utilization kinetic model for utilization of substrate were capable of predicting the fermentation profile with high coefficient of determination (R2) values of 0.980, 0.942 and 0.983 respectively. The results indicated that the unstructured models were able to describe the fermentation kinetics more effectively.

Keywords: Aspergillus flavus, Batch fermentation, Kinetic model, Tannase, Unstructured models.

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49 Immobilization of Aspergillus awamori 1-8 for Subsequent Pectinase Production

Authors: Zh. B. Suleimenova, Zh. K. Rakhmetova, R. K. Blieva, A. E. Nurlybayeva

Abstract:

The overall objective of this research is a strain improvement technology for efficient pectinase production. A novel cells cultivation technology by immobilization of fungal cells has been studied in long time continuous fermentations. Immobilization was achieved by using of new material for absorption of stores of immobilized cultures which was for the first time used for immobilization of microorganisms. Effects of various conditions of nitrogen and carbon nutrition on the biosynthesis of pectolytic enzymes in Aspergillus awamori 1-8 strain were studied. Proposed cultivation technology along with optimization of media components for pectinase overproduction led to increased pectinase productivity in Aspergillus awamori 1-8 from 7 to 8 times. Proposed technology can be applied successfully for production of major industrial enzymes such as α-amylase, protease, collagenase etc.

Keywords: Aspergillus awamori, immobilization, pectolytic enzymes.

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48 Production of Cellulases by Aspergillus Heteromorphus from Wheat Straw under Submerged Fermentation

Authors: Anita Singh, Namita Singh, Narsi R. Bishnoi

Abstract:

To investigate the production of cellulases from Aspergillus heteromorphus, submerged fermentation was performed using wheat straw as substrate. Optimization of saccharification conditions like pH, temperature and time were studied. Highest reducing sugar was released on 5th day at 5 pH, 30° C temperature. When A. heteromorphous was grown on wheat straw in submerged fermentation after 5 days incubation at 30 ° C, 3.2 IU/ml and 83 IU/ml, filter paper activity and CMCase activity respectively.

Keywords: Aspergillus heteromorphus, Wheat Straw, Submerged Fermentation, Production of Cellulases

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47 Fungal Leaching of Hazardous Heavy Metals from a Spent Hydrotreating Catalyst

Authors: R. Mafi Gholami, S. M. Borghei, S. M. Mousavi

Abstract:

In this study, the ability of Aspergillus niger and Penicillium simplicissimum to extract heavy metals from a spent refinery catalyst was investigated. For the first step, a spent processing catalyst from one of the oil refineries in Iran was physically and chemically characterized. Aspergillus niger and Penicillium simplicissimum were used to mobilize Al/Co/Mo/Ni from hazardous spent catalysts. The fungi were adapted to the mixture of metals at 100-800 mg L-1 with increments in concentration of 100 mg L-1. Bioleaching experiments were carried out in batch cultures. To investigate the production of organic acids in sucrose medium, analyses of the culture medium by HPLC were performed at specific time intervals after inoculation. The results obtained from Inductive coupled plasma-optical emission spectrometry (ICP-OES) showed that after the one-step bioleaching process using Aspergillus niger, maximum removal efficiencies of 27%, 66%, 62% and 38% were achieved for Al, Co, Mo and Ni, respectively. However, the highest removal efficiencies using Penicillium simplicissimum were of 32%, 67%, 65% and 38% for Al, Co, Mo and Ni, respectively

Keywords: Aspergillus niger, Bioleaching, Heavy metals, Penicillium simplicissimum, Spent catalyst

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46 Fungi Associated with Decline of Kikar (Acacia nilotica) and Red River Gum (Eucalyptus camaldulensis) in Faisalabad

Authors: I. Ahmad, A. Hannan, S. Ahmad, M. Asif, M. F. Nawaz, M. A. Tanvir, M. F. Azhar

Abstract:

During this research, a comprehensive survey of tree growing areas of Faisalabad district of Pakistan was conducted to observe the symptoms, spectrum, occurrence and severity of A. nilotica and E. camaldulensis decline. Objective of current research was to investigate specific fungal pathogens involved in decline of A. nilotica and E. camaldulensis. For this purpose, infected roots, bark, neck portion, stem, branches, leaves and infected soils were collected to identify associated fungi. Potato dextrose agar (PDA) and Czepak dox agar media were used for isolations. Identification of isolated fungi was done microscopically and different fungi were identified. During survey of urban locations of Faisalabad, disease incidence on Kikar and Eucalyptus was recorded as 3.9-7.9% and 2.6-7.1% respectively. Survey of Agroforest zones of Faisalabad revealed decline incidence on kikar 7.5% from Sargodha road while on Satiana and Jhang road it was not planted. In eucalyptus trees, 4%, 8% and 0% disease incidence was observed on Jhang road, Sargodha road and Satiana road respectively. The maximum fungus isolated from the kikar tree was Drechslera australiensis (5.00%) from the stem part. Aspergillus flavus also gave the maximum value of (3.05%) from the bark. Alternaria alternata gave the maximum value of (2.05%) from leaves. Rhizopus and Mucor spp. were recorded minimum as compared to the Drechslera, Alternaria and Aspergillus. The maximum fungus isolated from the Eucalyptus tree was Armillaria luteobubalina (5.00%) from the stem part. The other fungi isolated were Macrophamina phaseolina and A. niger.

Keywords: Decline, frequency of mycoflora, A. nilotica, E. camaldulensis, Drechslera australiensis, Armillaria luteobubalina.

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45 Kinetic Study of Gluconic Acid Batch Fermentation by Aspergillus niger

Authors: Akbarningrum Fatmawati, Rudy Agustriyanto, Lindawati

Abstract:

Gluconic acid is one of interesting chemical products in industries such as detergents, leather, photographic, textile, and especially in food and pharmaceutical industries. Fermentation is an advantageous process to produce gluconic acid. Mathematical modeling is important in the design and operation of fermentation process. In fact, kinetic data must be available for modeling. The kinetic parameters of gluconic acid production by Aspergillus niger in batch culture was studied in this research at initial substrate concentration of 150, 200 and 250 g/l. The kinetic models used were logistic equation for growth, Luedeking-Piret equation for gluconic acid formation, and Luedeking-Piret-like equation for glucose consumption. The Kinetic parameters in the model were obtained by minimizing non linear least squares curve fitting.

Keywords: Aspergillus niger, fermentation, gluconic acid, kinetic.

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44 Antidiabetic and Antioxidative Activities of Butyrolactone I from Aspergillus terreus MC751

Authors: Rizna Triana Dewi, Sanro Tachibana, Ahmad Darmawan

Abstract:

The bioassay-guided isolation and purification of an ethyl acetate extract of Aspergillus terreus MC751 led to the characterization of butyrolactone I as an antidiabetic and antioxidant. The antidiabetic activity of butyrolactone I was evaluated by α- glucosidase and α-amylase inhibition assays. Butyrolactone I demonstrated significant concentration-dependent, mixed-type inhibitory activity against yeast α-glucosidase with an IC50 of 54μM. However, the compound exhibited less activity against rat intestinal α-glucosidase and α-amylase. This is the first report on α-glucosidase inhibitory activity of butyrolactone I. The antioxidative activity of butyrolactone I was evaluated based on scavenging effects on 1,1- diphenyl-2-picrylhydrazyl (DPPH) (IC50 =51 μM) and hydrogen peroxide (IC50= 141 μM) radicals as well as a reducing power assay. The results suggest that butyrolactone I is a promising antidiabetic as well as antioxidant and should be considered for clinical trials.

Keywords: Aspergillus terreus MC751, antidiabetic, antioxidant, Butyrolactone I.

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43 Using Submerge Fermentation Method to Production of Extracellular Lipase by Aspergillus niger

Authors: Masoumeh Ghasemi, Afshin Farahbakhsh, Arman Farahbakhsh, Ali Asghar Safari

Abstract:

In this study, lipase production has been investigated using submerge fermentation by Aspergillus niger in Kilka fish oil as main substrate. The Taguchi method with an L9 orthogonal array design was used to investigate the effect of parameters and their levels on lipase productivity. The optimum conditions for Kilka fish oil concentration, incubation temperature and pH were obtained 3 gr./ml 35°C and 7, respectively. The amount of lipase activity in optimum condition was obtained 4.59IU/ml. By comparing this amount with the amount of productivity in the olive oil medium based on the cost of each medium, it was that using Kilka fish oil is 84% economical. Therefore Kilka fish oil can be used as an economical and suitable substrate in the lipase production and industrial usages.

Keywords: Lipase, Aspergillus niger, Kilka Fish oil, Submerge Fermentation method.

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42 In vitro Biological Activity of Some Synthesized Monoazo Heterocycles Based On Thiophene and Thiazolyl-Thiophene Analogues

Authors: M. E. Khalifa, A. A. Gobouri

Abstract:

Potential synthesis of a series of 3-amino-4-arylazothiophene derivatives from reaction of 2-cyano-2-phenylthiocarbamoyl acetamide and the appropriate α-halogenated reagents, followed by coupling with different aryl diazonium salts (Japp-Klingemann reaction), and another series of 5-arylazo-thiazol-2-ylcarbamoyl-thiophene derivatives from base-catalyzed intramolecular condensation of 5-arylazo-2-(N-chloroacetyl)amino-thiazole with selected b-keto compounds (Thorpe-Ziegler reaction) was performed. The biological activity of the two series was studied in vitro. Their versatility for pharmaceutical purposes was reported, where they displayed remarkable activities against selected pathogenic microorganisms; Bacillus subtilis, Staphylococcus aureus (Gram positive bacteria), Escherichia coli, Pseudomonas aeruginosa (Gram negative bacteria), and Aspergillus flavus, Candida albicans (fungi) with various degrees related to their chemical structures.

Keywords: 2-Aminothiazole, antimicrobial, monoazo compounds, thiophene, pathogenic microorganisms.

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41 Effects of Xylanase and Cellulase Production during Composting of EFB and POME using Fungi

Authors: Dayana Amira R., Roshanida A.R., Rosli M.I.

Abstract:

Empty Fruit Bunches (EFB) and Palm Oil Mill Effluent (POME) are two main wastes from oil palm industries which contain rich lignocellulose. Degradation of EFB and POME by microorganisms will produce hydrolytic enzyme which will degrade cellulose and hemicellulose during composting process. However, normal composting takes about four to six months to reach maturity. Hence, application of fungi into compost can shorten the period of composting. This study identifies the effect of xylanase and cellulase produced by Aspergillus niger and Trichoderma virens on composting process using EFB and POME. The degradation of EFB and POME indicates the lignocellulolytic capacity of Aspergillus niger and Trichoderma virens with more than 7% decrease in hemicellulose and more than 25% decrease in cellulose for both inoculated compost. Inoculation of Aspergillus niger and Trichoderma virens also increased the enzyme activities during the composting period compared to the control compost by 21% for both xylanase and cellulase. Rapid rise in the activities of cellulase and xylanase was observed by Aspergillus niger with the highest activities of 14.41 FPU/mg and 3.89 IU/mg, respectively. Increased activities of cellulase and xylanase also occurred in inoculation of Trichoderma virens with the highest activities obtained at 13.21 FPU/mg and 4.43 IU/mg, respectively. Therefore, it is evident that the inoculation of fungi can increase the enzyme activities hence effectively degrading the EFB and POME.

Keywords: EFB, cellulase, POME, xylanase

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40 Bioconversion of Oranges Wastes for Pectinase Production Using Aspergillus niger under Solid State Fermentation

Authors: N. Hachemi, A. Nouani, A. Benchabane

Abstract:

The influence of cultivation factors such as content of ammonium sulfate, glucose and water in the culture medium and particle size of dry orange waste, on their bioconversion for pectinase production was studied using complete factorial design. A polygalacturonase (PG) was isolated using ion exchange chromatography under gradient elution 0-0,5 m/l NaCl (column equilibrate with acetate buffer pH 4,5), subsequently by sephadex G75 column chromatography was applied and the molecular weight was obtained about 51,28 KDa. Purified PG enzyme exhibits a pH and temperature optima of activity at 5 and 35°C respectively. Treatment of apple juice by purified enzyme extract yielded a clear juice, which was competitive with juice yielded by pure Sigma Aldrich Aspergillus niger enzyme.

Keywords: Bioconversion, orange wastes, optimization, pectinase.

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39 Effect of Euphorbia Pulcherrima Leaf and Inflorescence Extract on Various Cytomorphological Parameters of Aspergillus fumigatus

Authors: Arti Goel, Kanika Sharma

Abstract:

Microorganisms can be removed, inhibited or killed by physical agents, physical processes or chemical agents but they have their inherent disadvantages such as increased resistance against antibiotics etc. Since, plants have endless ability to synthesize aromatic substances which act as the master agents for plant defense mechanisms against microorganisms, insects and herbivores. Thus, secondary metabolites or phytochemicals obtained from plants can be used as agents of disease control nowadays. In the present study effect of different concentrations of acetone fraction of leaves and alcohol fraction of inflorescence of Euphorbia pulcherrima on various cytomorphological parameters i.e. cell number, mycelium width, conidial size, conidiophore size etc. of Aspergillus fumigatus has been studied. Change in mycelium/ hyphal cell width, conidium size, conidiophore size etc. was measured with the help of a previously calibrated oculometer. To study effect on morphology, fungal mycelium along with conidiophore and conidia were stained with cotton blue and mounted in lactophenol and observed microscopically. Inhibitory action of the acetone extract of Euphorbia pulcherrima leaf on growth of Aspergillus fumigatus was investigated. Control containing extract free medium supported profuse growth of the fungus. Although decrease in growth was observed even at 3.95μg/ml but significant inhibition of growth was started at7.81μg/ml concentration of the extract. Complete inhibition was observed at 15.62μg/ml and above. Microscopic examination revealed that at 3.95, 7.81 and 15.62μg/ml extract concentration hyphal cell width was found to be increased from 1.44μm in control to 3.86, 5.24 and 8.98 μm respectively giving a beaded appearance to the mycelium. Vesicle size was reduced from 24.78x20.08μm (control) to 11.34x10.06μm at 3.95μg/ml concentration. At 7.81 and 15.62μg/ml concentration no phialides and sterigmata were observed. Inhibitory action of the alcohol extract of inflorescence on the growth of Aspergillus fumigatus was also studied. Control containing extract free medium supported profuse growth of the fungus. Although decrease in growth was observed even at 3.95μg/ml but complete inhibition was observed at 62.5μg/ml and above. Microscopic examination revealed that hyphal cell width of Aspergillus fumigatus was found to be increased from 1.67μm in control to 5.84μm at MIC i.e. at 62.5μg/ml. Vesicle size was reduced from 44.76x 24.22μm (control) to 11.36x 6.80μm at 15.62μg/ml concentrations. At 31.25 μg/ml and 62.5μg/ml concentration no phialides and sterigmata was found. Spore germination was completely found to be inhibited at 3.95μg/ml concentration. Similarly 92.87% reduction in vesicle size was observed at 15.62μg/ml concentration. It is evident from the results that plant extracts inhibit fungal growth and this inhibition is concentration dependent. 

Keywords: Antimicrobial Activity, Aspergillus fumigatus, cytomorphology, Euphorbia pulcherrima.

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38 Isolation and Molecular Identification of Two Fungal Strains Capable of Degrading Hydrocarbon Contaminants on Saudi Arabian Environment

Authors: Amr A. El Hanafy, Yasir Anwar, Saleh A. Mohamed, Saleh Mohamed Saleh Al-Garni, Jamal S. M. Sabir, Osama A. H. Abu Zinadah, Mohamed Morsi Ahmed

Abstract:

In the vicinity of red sea about 15 fungi species were isolated from oil contaminated sites. On the basis of aptitude to degrade the crude oil and DCPIP assay, two fungal isolates were selected amongst 15 oil degrading strains. Analysis of ITS-1, ITS-2 and amplicon pyrosequencing studies of fungal diversity revealed that these strains belong to Penicillium and Aspergillus species. Two strains that proved to be the most efficient in degrading crude oil was Aspergillus niger (54%) and Penicillium commune (48%) Subsequent to two weeks of cultivation in BHS medium the degradation rate were recorded by using spectrophotometer and GC-MS. Hence, it is cleared that these fungal strains has capability of degradation and can be utilize for cleaning the Saudi Arabian environment.

Keywords: Fungal strains, hydrocarbon contaminants, molecular identification, biodegradation, GC-MS.

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37 Cloning and Over Expression of an Aspergillus niger XP Phytase Gene (phyA) in Pichia pastoris

Authors: Ngo Thanh Xuan, Mai Thi Hang, Vu Nguyen Thanh

Abstract:

A. niger XP isolated from Vietnam produces very low amount of acidic phytase with optimal pH at 2.5 and 5.5. The phytase production of this strain was successfully improved through gene cloning and expression. A 1.4 - kb DNA fragment containing the coding region of the phyA gene was amplified by PCR and inserted into the expression vector pPICZαA with a signal peptide α- factor, under the control of AOX1 promoter. The recombined plasmid was transformed into the host strain P. pastoris KM71H and X33 by electroporation. Both host strains could efficiently express and secret phytase. The multicopy strains were screened for over expression of phytase. All the selected multicopy strains of P. pastoris X33 were examined for phytase activity, the maximum phytase yield of 1329 IU/ml was obtained after 4 days of incubation in medium BMM. The recombinant protein with MW of 97.4 KW showed to be the only one protein secreted in the culture broth. Multicopy transformant P. pastoris X33 supposed to be potential candidate for producing the commercial preparation of phytase.

Keywords: Aspergillus niger XP, cloning, over expression, Pichia pastoris, phyA , phytase.

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36 Microbial Assessment of Fenugreek Paste during Storage and Antimicrobial Effect of Greek Clover, Trigonella foenum-graecum

Authors: Zerrin Erginkaya, Gözde Konuray

Abstract:

In this study, antimicrobial effect of Greek clover was determined with usage of MIC (minimum inhibition concentration) and agar diffusion method. Moreover, pH, water activity and microbial change were determined during storage of fenugreek paste. At first part of our study, microbial load of spices was evaluated. Two different fenugreek pastes were produced with mixing of Greek clover, spices, garlic and water. Fenugreek pastes were stored at 4 °C. At the second part, antimicrobial effect of Greek clover was determined on Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Debaryomyces hansenii, Aspergillus parasiticus, Candida rugosa, Mucor spp., when the concentrations of Greek clover were 8%, 12% and 16%. According to the results obtained, mould growth was determined at 15th and 30th days of storage in first and second fenugreek samples, respectively. Greek clover showed only antifungal effect on Aspergillus parasiticus at previously mentioned concentrations.

Keywords: Antimicrobial, fenugreek, Greek clover, minimum inhibition concentration.

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35 Quantitative Structure Activity Relationship and Insilco Docking of Substituted 1,3,4-Oxadiazole Derivatives as Potential Glucosamine-6-Phosphate Synthase Inhibitors

Authors: Suman Bala, Sunil Kamboj, Vipin Saini

Abstract:

Quantitative Structure Activity Relationship (QSAR) analysis has been developed to relate antifungal activity of novel substituted 1,3,4-oxadiazole against Candida albicans and Aspergillus niger using computer assisted multiple regression analysis. The study has shown the better relationship between antifungal activities with respect to various descriptors established by multiple regression analysis. The analysis has shown statistically significant correlation with R2 values 0.932 and 0.782 against Candida albicans and Aspergillus niger respectively. These derivatives were further subjected to molecular docking studies to investigate the interactions between the target compounds and amino acid residues present in the active site of glucosamine-6-phosphate synthase. All the synthesized compounds have better docking score as compared to standard fluconazole. Our results could be used for the further design as well as development of optimal and potential antifungal agents.

Keywords: 1, 3, 4-Oxadiazole, QSAR, Multiple linear regression, Docking, Glucosamine-6-Phosphate Synthase.

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34 The Evaluation of Antioxidant and Antimicrobial Activities of Essential Oil and Aqueous, Methanol, Ethanol, Ethyl Acetate and Acetone Extract of Hypericum scabrum

Authors: A. Heshmati, M. Y Alikhani, M. T. Godarzi, M. R. Sadeghimanesh

Abstract:

Herbal essential oil and extracts are a good source of natural antioxidants and antimicrobial compounds. Hypericum is one of the potential sources of these compounds. In this study, the antioxidant and antimicrobial activity of essential oil and aqueous, methanol, ethanol, ethyl acetate and acetone extract of Hypericum scabrum was assessed. Flowers of Hypericum scabrum were collected from the surrounding mountains of Hamadan province and after drying in the shade, the essential oil of the plant was extracted by Clevenger and water, methanol, ethanol, ethyl acetate and acetone extract was obtained by maceration method. Essential oil compounds were identified using the GC-Mass. The Folin-Ciocalteau and aluminum chloride (AlCl3) colorimetric method was used to measure the amount of phenolic acid and flavonoids, respectively. Antioxidant activity was evaluated using DPPH and FRAP. The minimum inhibitory concentration (MIC) and the minimum bacterial/fungicide concentration (MBC/MFC) of essential oil and extracts were evaluated against Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Salmonella typhimurium, Aspergillus flavus and Candida albicans. The essential oil yield of was 0.35%, the lowest and highest extract yield was related to ethyl acetate and water extract. The most component of essential oil was α-Pinene (46.35%). The methanol extracts had the highest phenolic acid (95.65 ± 4.72 µg galic acid equivalent/g dry plant) and flavonoids (25.39 ± 2.73 µg quercetin equivalent/g dry plant). The percentage of DPPH radical inhibition showed positive correlation with concentrations of essential oil or extract. The methanol and ethanol extract had the highest DDPH radical inhibitory. Essential oil and extracts of Hypericum had antimicrobial activity against the microorganisms studied in this research. The MIC and MBC values for essential oils were in the range of 25-25.6 and 25-50 μg/mL, respectively. For the extracts, these values were 1.5625-100 and 3.125-100 μg/mL, respectively. Methanol extracts had the highest antimicrobial activity. Essential oil and extract of Hypericum scabrum, especially methanol extract, have proper antimicrobial and antioxidant activity, and it can be used to control the oxidation and inhibit the growth of pathogenic and spoilage microorganisms. In addition, it can be used as a substitute for synthetic antioxidant and antimicrobial compounds.

Keywords: Antimicrobial, antioxidant, extract, hypericum.

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33 A Risk Assessment Tool for the Contamination of Aflatoxins on Dried Figs based on Machine Learning Algorithms

Authors: Kottaridi Klimentia, Demopoulos Vasilis, Sidiropoulos Anastasios, Ihara Diego, Nikolaidis Vasileios, Antonopoulos Dimitrios

Abstract:

Aflatoxins are highly poisonous and carcinogenic compounds produced by species of the genus Aspergillus spp. that can infect a variety of agricultural foods, including dried figs. Biological and environmental factors, such as population, pathogenicity and aflatoxinogenic capacity of the strains, topography, soil and climate parameters of the fig orchards are believed to have a strong effect on aflatoxin levels. Existing methods for aflatoxin detection and measurement, such as high-performance liquid chromatography (HPLC), and enzyme-linked immunosorbent assay (ELISA), can provide accurate results, but the procedures are usually time-consuming, sample-destructive and expensive. Predicting aflatoxin levels prior to crop harvest is useful for minimizing the health and financial impact of a contaminated crop. Consequently, there is interest in developing a tool that predicts aflatoxin levels based on topography and soil analysis data of fig orchards. This paper describes the development of a risk assessment tool for the contamination of aflatoxin on dried figs, based on the location and altitude of the fig orchards, the population of the fungus Aspergillus spp. in the soil, and soil parameters such as pH, saturation percentage (SP), electrical conductivity (EC), organic matter, particle size analysis (sand, silt, clay), concentration of the exchangeable cations (Ca, Mg, K, Na), extractable P and trace of elements (B, Fe, Mn, Zn and Cu), by employing machine learning methods. In particular, our proposed method integrates three machine learning techniques i.e., dimensionality reduction on the original dataset (Principal Component Analysis), metric learning (Mahalanobis Metric for Clustering) and K-nearest Neighbors learning algorithm (KNN), into an enhanced model, with mean performance equal to 85% by terms of the Pearson Correlation Coefficient (PCC) between observed and predicted values.

Keywords: aflatoxins, Aspergillus spp., dried figs, k-nearest neighbors, machine learning, prediction

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32 Novel Dual Stage Membrane Bioreactor for the Continuous Remediation of Electroplating Wastewater

Authors: B. A. Q. Santos, S. K. O. Ntwampe, G. Muchatibaya

Abstract:

In this study, the designed dual stage membrane bioreactor (MBR) system was conceptualized for the treatment of cyanide and heavy metals in electroplating wastewater. The design consisted of a primary treatment stage to reduce the impact of fluctuations and the secondary treatment stage to remove the residual cyanide and heavy metal contaminants in the wastewater under alkaline pH conditions. The primary treatment stage contained hydrolyzed Citrus sinensis (C. sinensis) pomace and the secondary treatment stage contained active Aspergillus awamori (A. awamori) biomass, supplemented solely with C. sinensis pomace extract from the hydrolysis process. An average of 76.37%, 95.37%, 93.26 and 94.76% and 99.55%, 99.91%, 99.92% and 99.92% degradation efficiency for total cyanide (T-CN), including the sorption of nickel (Ni), zinc (Zn) and copper (Cu) were observed after the first and second treatment stages, respectively. Furthermore, cyanide conversion by-products degradation was 99.81% and 99.75 for both formate (CHOO-) and ammonium (NH4 +) after the second treatment stage. After the first, second and third regeneration cycles of the C. sinensis pomace in the first treatment stage, Ni, Zn and Cu removal achieved was 99.13%, 99.12% and 99.04% (first regeneration cycle), 98.94%, 98.92% and 98.41% (second regeneration cycle) and 98.46 %, 98.44% and 97.91% (third regeneration cycle), respectively. There was relatively insignificant standard deviation detected in all the measured parameters in the system which indicated reproducibility of the remediation efficiency in this continuous system.

Keywords: Aspergillus awamori, Citrus sinensis pomace, electroplating wastewater remediation, membrane bioreactor.

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31 Inulinase Immobilization on Functionalized Magnetic Nanoparticles Prepared with Soy Protein Isolate Conjugated Bovine Serum Albumin for High Fructose Syrup Production

Authors: Homa Torabizadeh, Mohaddeseh Mikani

Abstract:

Inulinase from Aspergillus niger was covalently immobilized on magnetic nanoparticles (MNPs/Fe3O4) covered with soy protein isolate (SPI/Fe3O4) functionalized by bovine serum albumin (BSA) nanoparticles. MNPs are promising enzyme carriers because they separate easily under external magnetic fields and have enhanced immobilized enzyme reusability. As MNPs aggregate simply, surface coating strategy was employed. SPI functionalized by BSA was a suitable candidate for nanomagnetite coating due to its superior biocompatibility and hydrophilicity. Fe3O4@SPI-BSA nanoparticles were synthesized as a novel carrier with narrow particle size distribution. Step by step fabrication monitoring of Fe3O4@SPI-BSA nanoparticles was performed using field emission scanning electron microscopy and dynamic light scattering. The results illustrated that nanomagnetite with the spherical morphology was well monodispersed with the diameter of about 35 nm. The average size of the SPI-BSA nanoparticles was 80 to 90 nm, and their zeta potential was around −34 mV. Finally, the mean diameter of fabricated Fe3O4@SPI-BSA NPs was less than 120 nm. Inulinase enzyme from Aspergillus niger was covalently immobilized through gluteraldehyde on Fe3O4@SPI-BSA nanoparticles successfully. Fourier transform infrared spectra and field emission scanning electron microscopy images provided sufficient proof for the enzyme immobilization on the nanoparticles with 80% enzyme loading.

Keywords: High fructose syrup, inulinase immobilization, functionalized magnetic nanoparticles, soy protein isolate.

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30 Synthesis, Physicochemical Characterization and Study of the Antimicrobial Activity of Chlorobutanol

Authors: N. Hadhoum, B. Guerfi, T. M. Sider, Z. Yassa, T. Djerboua, M. Boursouti, M. Mamou, F. Z. Hadjadj Aoul, L. R. Mekacher

Abstract:

Introduction and objectives: Chlorobutanol is a raw material, mainly used as an antiseptic and antimicrobial preservative in injectable and ophthalmic preparations. The main objective of our study was the synthesis and evaluation of the antimicrobial activity of chlorobutanol hemihydrates. Material and methods: Chlorobutanol was synthesized according to the nucleophilic addition reaction of chloroform to acetone, identified by an infrared absorption using Spectrum One FTIR spectrometer, melting point, Scanning electron microscopy and colorimetric reactions. The dosage of carvedilol active substance was carried out by assaying the degradation products of chlorobutanol in a basic solution. The chlorobutanol obtained was subjected to bacteriological tests in order to study its antimicrobial activity. The antibacterial activity was evaluated against strains such as Escherichia coli (ATCC 25 922), Staphylococcus aureus (ATCC 25 923) and Pseudomonas aeroginosa (ATCC = American type culture collection). The antifungal activity was evaluated against human pathogenic fungal strains, such as Candida albicans and Aspergillus niger provided by the parasitology laboratory of the Hospital of Tizi-Ouzou, Algeria. Results and discussion: Chlorobutanol was obtained in an acceptable yield. The characterization tests of the product obtained showed a white and crystalline appearance (confirmed by scanning electron microscopy), solubilities (in water, ethanol and glycerol), and a melting temperature in accordance with the requirements of the European pharmacopoeia. The colorimetric reactions were directed towards the presence of a trihalogenated carbon and an alcohol function. The spectral identification (IR) showed the presence of characteristic chlorobutanol peaks and confirmed the structure of the latter. The microbiological study revealed an antimicrobial effect on all strains tested (Sataphylococcus aureus (MIC = 1250 µg/ml), E. coli (MIC = 1250 µg/ml), Pseudomonas aeroginosa (MIC = 1250 µg/ml), Candida albicans (MIC =2500 µg/ml), Aspergillus niger (MIC =2500 µg/ml)) with MIC values close to literature data. Conclusion: Thus, on the whole, the synthesized chlorobutanol satisfied the requirements of the European Pharmacopoeia, and possesses antibacterial and antifungal activity; nevertheless, it is necessary to insist on the purification step of the product in order to eliminate the maximum impurities.

Keywords: Antimicrobial agent, bacterial and fungal strains, chlorobutanol, MIC.

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29 Synthesis, Physicochemical Characterization and Study of the Antimicrobial Activity of Chlorobutanol

Authors: H. Nadia, G. Bahdja, S. Thili Malha, Y. Zahoua, D. Taoufik, B. Mourad, M. Marzouk, F. Z. Hadjadj Aoul, L. R. Mekacher

Abstract:

Introduction and objectives: Chlorobutanol is a raw material, mainly used as an antiseptic and antimicrobial preservative in injectable and ophthalmic preparations. The main objective of our study was the synthesis and evaluation of the antimicrobial activity of chlorobutanol hemihydrates. Material and methods: Chlorobutanol was synthesized according to the nucleophilic addition reaction of chloroform to acetone, identified by an infrared absorption using Spectrum One FTIR spectrometer, melting point, Scanning electron microscopy and colorimetric reactions. The dosage of Carvedilol active substance was carried out by assaying the degradation products of chlorobutanol in a basic solution. The chlorobutanol obtained was subjected to bacteriological tests in order to study its antimicrobial activity. The antibacterial activity was evaluated against strains such as Escherichia coli (ATCC 25 922), Staphylococcus aureus (ATCC 25 923) and Pseudomonas aeroginosa (ATCC = American type culture collection). The antifungal activity was evaluated against human pathogenic fungal strains, such as Candida albicans and Aspergillus niger provided by the parasitology laboratory of the Hospital of Tizi-Ouzou, Algeria. Results and discussion: Chlorobutanol was obtained in an acceptable yield. The characterization tests of the product obtained showed a white and crystalline appearance (confirmed by scanning electron microscopy), solubilities (in water, ethanol and glycerol), and a melting temperature in accordance with the requirements of the European pharmacopoeia. The colorimetric reactions were directed towards the presence of a trihalogenated carbon and an alcohol function. The spectral identification (IR) showed the presence of characteristic chlorobutanol peaks and confirmed the structure of the latter. The microbiological study revealed an antimicrobial effect on all strains tested (Sataphylococcus aureus (MIC = 1250 µg/ml), E. coli (MIC = 1250 µg/ml), Pseudomonas aeroginosa (MIC = 1250 µg/ml), Candida albicans (MIC =2500 µg/ml), Aspergillus niger (MIC =2500 µg/ml)) with MIC values close to literature data. Conclusion: Thus, on the whole, the synthesized chlorobutanol satisfied the requirements of the European Pharmacopoeia, and possesses antibacterial and antifungal activity; nevertheless it is necessary to insist on the purification step of the product in order to eliminate the maximum impurities.

Keywords: Antimicrobial agent, bacterial and fungal strains, chlorobutanol, MIC.

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28 Potential of Agro-Waste Extracts as Supplements for the Continuous Bioremediation of Free Cyanide Contaminated Wastewater

Authors: Seteno K. O. Ntwampe, Bruno A. Q. Santos

Abstract:

Different agricultural waste peels were assessed for their suitability to be used as primary substrates for the bioremediation of free cyanide (CN-) by a cyanide-degrading fungus Aspergillus awamori isolated from cyanide containing wastewater. The bioremediated CN- concentration were in the range of 36 to 110 mg CN-/L, with Orange (C. sinensis) > Carrot (D. carota) > Onion (A. cepa) > Apple (M. pumila), being chosen as suitable substrates for large scale CN- degradation processes due to: 1) the high concentration of bioremediated CN-, 2) total reduced sugars released into solution to sustain the biocatalyst, and 3) minimal residual NH4- N concentration after fermentation. The bioremediation rate constants (k) were 0.017h-1 (0h < t < 24h), with improved bioremediation rates (0.02189h-1) observed after 24h. The averaged nitrilase activity was ~10 U/L.

Keywords: Agricultural waste, Bioremediation, Cyanide, Wastewater.

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27 Feasibility Study on Vanillin Production from Jatropha curcas Stem Using Steam Explosion as a Pretreatment

Authors: Pilanee Vaithanomsat, Waraporn Apiwatanapiwat

Abstract:

Jatropha curcas stem was analyzed for chemical compositions: 19.11% pentosan, 42.99% alphacellulose and 24.11% lignin based on dry weight of 100-g raw material. The condition to fractionate cellulose, hemicellulose and lignin in J. curcas stem using steam explosion was optimized. The procedure started from cutting J. curcas stem into small pieces and soaked in water for overnight. After that, they were steam exploded at 214 °C and 21 kg/cm2 for 5 min. The obtained hydrolysate contained 1.55 g/L ferulic acid which after that was used as substrate for vanillin production by Aspergillus niger and Pycnoporus cinnabarinus in one-step process. The maximum 0.65 g/L of vanillin were obtained with the conversion rate of 45.2% based on the initial ferulic acid.

Keywords: Vanillin, production, Jatropha curcas stem, steam explosion.

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