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

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

Preparation and in vivo Assessment of Nystatin-Loaded Solid Lipid Nanoparticles for Topical Delivery against Cutaneous Candidiasis

Solid lipid nanoparticles (SLNs) have gained great attention for the topical treatment of skin associated fungal infection as they facilitate the skin penetration of loaded drugs. Our work deals with the preparation of nystatin loaded solid lipid nanoparticles (NystSLNs) using the hot homogenization and ultrasonication method. The prepared NystSLNs were characterized in terms of entrapment efficiency, particle size, zeta potential, transmission electron microscopy, differential scanning calorimetry, rheological behavior and in vitro drug release. A stability study for 6 months was performed. A microbiological study was conducted in male rats infected with Candida albicans, by counting the colonies and examining the histopathological changes induced on the skin of infected rats. The results showed that SLNs dispersions are spherical in shape with particle size ranging from 83.26±11.33 to 955.04±1.09 nm. The entrapment efficiencies are ranging from 19.73±1.21 to 72.46±0.66% with zeta potential ranging from -18.9 to -38.8 mV and shear-thinning rheological Behavior. The stability studies done for 6 months showed that nystatin (Nyst) is a good candidate for topical SLN formulations. A least number of colony forming unit/ ml (cfu/ml) was recorded for the selected NystSLN compared to the drug solution and the commercial Nystatin® cream present in the market. It can be fulfilled from this work that SLNs provide a good skin targeting effect and may represent promising carrier for topical delivery of Nyst offering the sustained release and maintaining the localized effect, resulting in an effective treatment of cutaneous fungal infection.

Levels of Some Antinutritional Factors in Tempeh Produced From Some Legumes and Jojobas Seeds

Three legumes i.e. soybean, kidney bean and mung bean, and jojoba seed as an oil seed were processed into tempeh, a fermented food. Changes in phytic acid, total phenols and trypsin inhibitor were monitored during the pretreatments (soaking, soaking– dehulling, washing and cooking) and fermentation with Rhizopus oligosporus. Soaking was found to reduce total phenol and trypsin inhibitor levels in soybean, kidney bean and mung bean. However, phytic acid was reduced by soaking in kidney bean and mung bean. Cooking was the most effective in reducing the activity of trypsin inhibitor. During fermentation, a slight increase in the level of trypsin inhibitor was noticed in soybean. Phytic acid and total phenols were decreased during fermentation in soybean, kidney bean but mung bean faild to form tempeh because the antifungal activity of herein a protein in mung bean, which exerts both chitinase activity and antifungal activity against a variety of fungal species. On the other hand, solid- state fermentation of jojoba seeds was not effective in reducing their content from cyanogenic glycosides (simmondsin).

Influence of Fermentation Conditions on Humic Acids Production by Trichoderma viride Using an Oil Palm Empty Fruit Bunch as the Substrate

Humic acids (HA) were produced by a Trichoderma viride strain under submerged fermentation in a medium based on the oil palm empty fruit bunch (EFB) and the main variables of the process were optimized by using response surface methodology. A temperature of 40°C and concentrations of 50g/L EFB, 5.7g/L potato peptone and 0.11g/L (NH4)2SO4 were the optimum levels of the variables that maximize the HA production, within the physicochemical and biological limits of the process. The optimized conditions led to an experimental HA concentration of 428.4±17.5 mg/L, which validated the prediction from the statistical model of 412.0mg/L. This optimization increased about 7–fold the HA production previously reported in the literature. Additionally, the time profiles of HA production and fungal growth confirmed our previous findings that HA production preferably occurs during fungal sporulation. The present study demonstrated that T. viride successfully produced HA via the submerged fermentation of EFB and the process parameters were successfully optimized using a statistics-based response surface model. To the best of our knowledge, the present work is the first report on the optimization of HA production from EFB by a biotechnological process, whose feasibility was only pointed out in previous works.

Development of Efficient Fungal Biomass-Degrading Enzyme Mixtures for Saccharification of Local Lignocellulosic Feedstock

Conversion of lignocellulosic biomass is the basis process for production of fuels, chemicals and materials in the sustainable biorefinery industry. Saccharification of lignocellulosic biomass is an essential step which produces sugars for further conversion to target value-added products e.g. bio-ethanol, bio-plastic, g-valerolactone (GVL), 5-hydroxymethylfuroic acid (HMF), levulinic acid, etc. The goal of this work was to develop an efficient enzyme for conversion of biomass to reducing sugar based on crude fungal enzyme from Chaetomium globosum BCC5776 produced by submerged fermentation and evaluate its activity comparing to a commercial Acremonium cellulase. Five local biomasses in Thailand: rice straw, sugarcane bagasse, corncobs, corn stovers, and palm empty fruit bunches were pretreated and hydrolyzed with varying enzyme loadings. Saccharification of the biomass led to different reducing sugar levels from 115 mg/g to 720 mg/g from different types of biomass using cellulase dosage of 9 FPU/g. The reducing sugar will be further employed as sugar feedstock for production of ethanol or commodity chemicals. This work demonstrated the use of promising enzyme candidate for conversion of local lignocellulosic biomass in biorefinery industry.

Preparation of Tempeh Spore Powder by Freeze Drying

Study production of tempeh inoculums powder by freeze-drying comparison with dry at 50°C and the sun bask for developing efficient tempeh inoculums for tempeh producing. Rhizopus oligosporus in PDA slant cultures was incubated at 30oC for 3-5 days until spores and mycelium. Preparation spores suspension with sterilized water and then count the number of started spores. Fill spores suspension in Rice flour and soy flour, mixed with water (in the ratio 10: 7), which is steamed and sterilized at 121°C 15min. Incubated at room temperature for 4 days, count number of spores. Then take the progressive infection and full spore dough to dry at 50°C, sun bask, and lyophilize. Grind to powder. Then pack in plastic bags, stored at 5°C. To investigate quality of inoculums which use different methods, tempeh was fermented every 4 weeks for 24 weeks of the experiment. The result found that rice flour is not suitable to use as raw material in the production of powdered spores.  Fungi can growth rarely. Less number of spores and requires more time than soy flour. For drying method, lyophilization is the least possible time. Samples from this method are very hard and very dark and harder to grind than other methods. Drying at 50°C takes longer time than lyophilization but can also set time use for drying. Character of the dry samples is hard solid and brown color, but can be grinded easier. The sun drying takes the longest time, can’t determine the exact time. When the spore powder was used to fermented tempeh immediately, product has similar characters as which use spores that was fresh prepared. The tempeh has normal quality. When spore powder stored at low temperature, tempeh from storage spore in weeks 4, 8 and 12 is still normal. Time spending in production was close to the production of fresh spores. After storage  spores for 16 and 20 weeks, tempeh is still normal but growth and sporulation were take longer time than usual (about 6 hours). At 24 week storage, fungal growth is not good, made tempeh looks inferior to normal color, also smell and texture.

Antimicrobial Effect of Essential Oil of Plant Schinus molle on Some Bacteria Pathogens

Humans use plants for thousands of years to treat various ailments, in many developing countries; much of the population relies on traditional doctors and their collections of medicinal plants to cure them. Essential oils have many therapeutic properties. In herbal medicine, they are used for their antiseptic properties against infectious diseases of fungal origin, against dermatophytes, those of bacterial origin. The aim of our study is to determine the antimicrobial effect of essential oils of the plant Schinus molle on some pathogenic bacteria. It is a medicinal plant used in traditional therapy. Essential oils have many therapeutic properties. In herbal medicine, they are used for their antiseptic properties against infectious diseases of fungal origin, against dermatophytes, those of bacterial origin. The test adopted, is based on the diffusion method on solid medium (Antibiogram), this method allows to determine the susceptibility or resistance of an organism according to the sample studied. Our study reveals that the essential oil of the plant Schinus molle has a different effect on the resistance of germs: for Pseudomonas aeruginosa strain is a moderately sensitive with an inhibition zone of 10mm, further Enterobacter, Escherichia coli and Proteus are strains that represent a high sensitivity, a zone of inhibition equal to 14.66 mm.

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

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.

Microbial Evaluation of Geophagic and Cosmetic Clays from Southern and Western Nigeria: Potential Natural Nanomaterials

Geophagic and cosmetic clays are among potential nanomaterial which occur naturally and are of various forms. The use of these nanoclays is a common practice in both rural and urban areas mostly due to tradition and medicinal reasons. These naturally occurring materials can be valuable sources of nanomaterial by serving as nanocomposites. The need to ascertain the safety of these materials is the motivation for this research. Physical Characterization based on the hue value and microbiological qualities of the nanoclays were carried out. The Microbial analysis of the clay samples showed considerable contamination with both bacteria and fungi with fungal contaminants taking the lead. This observation may not be unlikely due to the ability of fungi species to survive harsher growth conditions than bacteria. ‘Atike pupa’ showed no bacterial growth. The clay with the largest bacterial count was Calabash chalk (Igbanke), while that with the highest fungal count was ‘Eko grey’. The most commonly isolated bacteria in this study were Clostridium spp. and Corynebacterium spp. while fungi included Aspergillus spp. These results are an indication of the need to subject these clay materials to treatments such as heating before consumption or topical usage thereby ascertaining their safety.

Antimicrobial Activity of Girardinia heterophylla

In the present study an attempt has been made to prepare the crude extracts of leaves and stem of ‘Girardinia heterophylla’ by using various solvents like petroleum ether, ethanol and double distilled water. The samples were given the code NGLS 1, NGLS 2, NGLS 3 and NGSS 1, NGSS 2 and NGSS 3 respectively. All the extracts were used to study their antimicrobial activity against gram positive bacteria e.g. Bacillus subtilis, gram negative bacteria e.g. E. coli and K. pneumonia and antifungal activity against Aspergillus niger. The results of the antimicrobial activity showed that all the crude extracts of the plant possesses antibacterial activity. Maximum antibacterial activity was shown by NGLS 2, NGLS 3 and NGSS 3 against K. pneumonia. The growth of fungus A. niger was also inhibited by all the crude extracts. Maximum inhibition was shown by NGSS 2 followed by NGSS 1.

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

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. 

Use of Multiple Linear Regressions to Evaluate the Influence of O3 and PM10 on Biological Pollutants

Exposure to ambient air pollution has been linked to a number of health outcomes, starting from modest transient changes in the respiratory tract and impaired pulmonary function, continuing to restrict activity/reduce performance and to the increase emergency rooms visits, hospital admissions or mortality. The increase of allergenic symptoms has been associated with air contaminants such as ozone, particulate matter, fungal spores and pollen. Considering the potential relevance of crossed effects of nonbiological pollutants and airborne pollens and fungal spores on allergy worsening, the aim of this work was to evaluate the influence of non-biological pollutants (O3 and PM10) and meteorological parameters on the concentrations of pollen and fungal spores using multiple linear regressions. The data considered in this study were collected in Oporto which is the second largest Portuguese city, located in the North. Daily mean of O3, PM10, pollen and fungal spore concentrations, temperature, relative humidity, precipitation, wind velocity, pollen and fungal spore concentrations, for 2003, 2004 and 2005 were considered. Results showed that the 90th percentile of the adjusted coefficient of determination, P90 (R2aj), of the multiple regressions varied from 0.613 to 0.916 for pollen and from 0.275 to 0.512 for fungal spores. O3 and PM10 showed to have some influence on the biological pollutants. Among the meteorological parameters analysed, temperature was the one that most influenced the pollen and fungal spores airborne concentrations. Relative humidity also showed to have some influence on the fungal spore dispersion. Nevertheless, the models for each pollen and fungal spore were different depending on the analysed period, which means that the correlations identified as statistically significant can not be, even so, consistent enough.

Antibacterial and Antifungal Activity Assesment of Nigella Sativa Essential Oils

Antifungal activities of ether and methanolic extracts of volatiles oils of Nigella Sativa seeds were tested against pathogenic bacterias and fungies strains.The volatile oil were found to have significant antifungal and antibacterial activities compare to tetracycline, cefuroxime and ciprofloxacin positive controls.The ether and methanolic esxtracts were compared to each other for antifungal and antibacterial activities and ether extracts showed stonger activity than methanolic one.

Bioprocessing of Proximally Analyzed Wheat Straw for Enhanced Cellulase Production through Process Optimization with Trichodermaviride under SSF

The purpose of the present work was to study the production and process parameters optimization for the synthesis of cellulase from Trichoderma viride in solid state fermentation (SSF) using an agricultural wheat straw as substrates; as fungal conversion of lignocellulosic biomass for cellulase production is one among the major increasing demand for various biotechnological applications. An optimization of process parameters is a necessary step to get higher yield of product. Several kinetic parameters like pretreatment, extraction solvent, substrate concentration, initial moisture content, pH, incubation temperature and inoculum size were optimized for enhanced production of third most demanded industrially important cellulase. The maximum cellulase enzyme activity 398.10±2.43 μM/mL/min was achieved when proximally analyzed lignocellulosic substrate wheat straw inocubated at 2% HCl as pretreatment tool along with distilled water as extraction solvent, 3% substrate concentration 40% moisture content with optimum pH 5.5 at 45°C incubation temperature and 10% inoculum size.

Effect of Butachlor on the Microbial Population of Direct Sown Rice

Field experiments were conducted at Annamalai University Experimental Farm, Department of Agronomy; to device suitable weed control measures for direct seeded puddled rice and to study the effect of the weed control measures on the soil microbial population. The treatments comprised of incorporation of pressmud @ 6.25 t ha-1 and application of herbicide butachlor @1.5 kg a. i. ha- 1 with and without safener 4 days after sowing (DAS), 8 DAS alone and also in conjunction with hand weeding at 30 DAS. Hand weeding twice and a weedy check were also maintained. At maximum tillering stage, the population of bacteria was significantly reduced by butachlor application. The injury to microbes caused by herbicide disappeared with the advancement of crop's age and at flowering stage of crop, there was no significant difference among the treatments. The fungal and actinomycetes population remained unaltered by weed control treatments at both the stages of observation.

Pt(IV) Complexes with Polystrene-bound Schiff Bases as Antimicrobial Agent: Synthesis and Characterization

Novel polystrene-bound Schiff bases and their Pt(IV) complexes have been prepared from condensation reaction of polystyrene-A-NH2 with 2-hydroxybenzaldehyde and 5-fluoro-3- bromo-2-hydroxybenzaldehyde. The structures of Pt(IV) complexes with polystyrene including Schiff bases have been determined by elemental analyses, magnetic susceptibility, IR, 1H-NMR, UV-vis, TG/DTA and AAS. The antibacterial and antifungal activities of the synthesized compounds have been studied by the well-diffusion method against some selected microorganisms: (Bacillus cereus spp., Listeria monocytogenes 4b, Micrococcus luteus, Staphylococcus aureus, Staphylococcus epidermis, Brucella abortus, Escherichia coli, Pseudomonas putida spp., Shigella dysenteria type 10, Salmonella typhi H).

Evaluation of Antifungal Potential of Cenchrus pennisetiformis for the Management of Macrophomina phaseolina

Macrophomina phaseolina is a devastating soil-borne fungal plant pathogen that causes charcoal rot disease in many economically important crops worldwide. So far, no registered fungicide is available against this plant pathogen. This study was planned to examine the antifungal activity of an allelopathic grass Cenchrus pennisetiformis (Hochst. & Steud.) Wipff. for the management of M. phaseolina isolated from cowpea [Vigna unguiculata (L.) Walp.] plants suffering from charcoal rot disease. Different parts of the plants viz. inflorescence, shoot and root were extracted in methanol. Laboratory bioassays were carried out using different concentrations (0, 0.5, 1.0, …, 3.0 g mL-1) of methanolic extracts of the test allelopathic grass species to assess the antifungal activity against the pathogen. In general, extracts of all parts of the grass exhibited antifungal activity. All the concentrations of methanolic extracts of shoot and root significantly reduced fungal biomass by 20–73% and 40–80%, respectively. Methanolic shoot extract was fractionated using n-hexane, chloroform, ethyl acetate and n-butanol. Different concentrations of these fractions (3.125, 6.25, …, 200 mg mL-1) were analyzed for their antifungal activity. All the concentrations of n-hexane fraction significantly reduced fungal biomass by 15–96% over corresponding control treatments. Higher concentrations (12.5–200 mg mL-1) of chloroform, ethyl acetate and n-butanol also reduced the fungal biomass significantly by 29–100%, 46–100% and 24–100%, respectively.

Inhibition of the Growth of Pathogenic Candida spp. by Salicylhydroxamic Acid

Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth each of six Candida spp. was inhibited significantly by ~13 mM SHAM or 2 mM cyanide, albeit to differing extents. In C. dubliniensis, C. krusei and C. tropicalis the rate of O2 uptake was inhibited by 18-36% by 25 mM SHAM, but this had little or no effect on C. glabrata, C. guilliermondii or C. parapsilosis. Although SHAM substantially inhibited the growth of Candida spp., it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of Candida spp. infection.

Antimicrobial Effect of Essential oil of Plant Trigonella focnum greacum on some Bacteria Pathogens

The plant world is the source of many medicines. Recently, researchers have estimated that there are approximately 400,000 plant species worldwide, of which about a quarter or a third have been used by societies for medicinal purposes. The human uses of plants for thousands of years to treat various ailments, in many developing countries, much of the population trust in traditional doctors and their collections of medicinal plants to treat them. Essential oils have many therapeutic properties. In herbal medicine, they are used for their antiseptic properties against infectious diseases of fungal origin, against dermatophytes, those of bacterial origin. The aim of our study is to determine the antimicrobial effect of essential oils of the plant Trigonella focnum greacum on some pathogenic bacteria, it is a medicinal plant used in traditional therapy. The test adopted is based on the diffusion method on solid medium (Antibiogram), this method determines the sensitivity or resistance of a microorganism vis-à-vis the extract studied. Our study reveals that the essential oil of the plant Trigonella focnum greacum has a different effect on the resistance of germs. For staphiloccocus Pseudomonnas aeroginosa and Krebsilla, are moderately sensitive strains, also Escherichia coli and Candida albicans represents a high sensitivity. By against Proteus is a strain that represents a weak sensitivity.

Antifungal Activity of Silver Colloidal Nanoparticles against Phytopathogenic Fungus (Phomopsis sp.) in Soybean Seeds

Among the many promising nanomaterials with antifungal properties, metal nanoparticles (silver nanoparticles) stand out due to their high chemical activity. Therefore, the aim of this study was to evaluate the effect of silver nanoparticles (AgNPs) against Phomopsis sp. AgNPs were synthesized by silver nitrate reduction with sodium citrate and stabilized with ammonia. The synthesized AgNPs have further been characterized by UV/Visible spectroscopy, Biophysical techniques like Dynamic light scattering (DLS) and Scanning Electron Microscopy (SEM). The average diameter of the prepared silver colloidal nanoparticles was about 52 nm. Absolute inhibitions (100%) were observed on treated with a 270 and 540 µg ml-1 concentration of AgNPs. The results from the study of the AgNPs antifungal effect are significant and suggest that the synthesized silver nanoparticles may have an advantage compared with conventional fungicides.