Search results for: entomopathogenic fungi

Authors: Patarasuda Chaisupa, R. Clay Wright

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The selection of appropriate biomolecular targets is a crucial aspect of biopharmaceutical development. The Auxin-Inducible Degron Degradation (AID) technology has demonstrated remarkable potential in efficiently and rapidly degrading target proteins, thereby enabling the identification and acquisition of drug targets. The AID system also offers a viable method to deplete specific proteins, particularly in cases where the degradation pathway has not been exploited or when the adaptation of proteins, including the cell environment, occurs to compensate for the mutation or gene knockout. In this study, we have engineered an improved AID system tailored to deplete proteins of interest. This AID construct combines the auxin-responsive E3 ubiquitin ligase binding domain, AFB2, and the substrate degron, IAA17, fused to the target genes. Essential genes of fungi with the lowest percent amino acid similarity to human and plant orthologs, according to the Basic Local Alignment Search Tool (BLAST), were cloned into the AID construct in S. cerevisiae (AID-tagged strains) using a modular yeast cloning toolkit for multipart assembly and direct genetic modification. Each E3 ubiquitin ligase and IAA17 degron was fused to a fluorescence protein, allowing for real-time monitoring of protein levels in response to different auxin doses via cytometry. Our AID system exhibited high sensitivity, with an EC50 value of 0.040 µM (SE = 0.016) for AFB2, enabling the specific promotion of IAA17::target protein degradation. Furthermore, we demonstrate how this improved AID system enhances quantitative functional studies of various proteins in fungi. The advancements made in auxin-inducible protein degradation in this study offer a powerful approach to investigating critical target protein viability in fungi, screening protein targets for drugs, and regulating intracellular protein abundance, thus revolutionizing the study of protein function underlying a diverse range of biological processes.

Keywords: synthetic biology, bioengineering, molecular biology, biotechnology

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Authors: Ezeonu, Chukwuma Stephen, Onwurah, Ikechukwu Noel Emmanuel

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Pure strains of Aspergillus fumigatus (AF), aspergillus niger (AN), aspergillus oryzae (AO), trichophyton mentagrophyte (TM), trichophyton rubrum (TR) and Trichophyton soudanense (TS) were isolated from decomposing rice husk. Freshly processed rice husk in Mandle’s medium were heat pre-treated using an autoclave at 121oC for 20 minutes. The isolated fungi as monoculture and di-culture combinations were inoculated into each of the pre-treated rice husk with the exception of two controls. Seven days hydrolysis was followed by estimation of carbohydrate, reducing sugar and non-reducing sugar. Fungal treated rice husks were left to ferment for 7 days with introduction of both baker’s and palm wine yeast. The result obtained in the work gave the highest carbohydrate (20.53 ± 2.73 %) from rice husks treated with TS + TR di-culture. The highest soluble reducing sugar (2.66 ± 0.14 %) was obtained from rice husk treated with TM. The highest soluble nonreducing sugar (18.08 ± 2.61 %) was from AF. The introduction of yeasts from palm wine gave the highest bio-ethanol (12.82 ± 0.39 %) from AO. The highest bio-ethanol (6.60 ± 0.10 %) from baker's yeast fermentation was in AO + TS treated rice husk. There was increased availability of sugar and moderate yield of bio-ethanol, especially from palm wine yeast.

Keywords: fungi, rice husk, carbohydrate, reducing sugar, non-reducing sugar, ethanol, fermentation

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Authors: Musa Filibus Gugu

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An investigation was carried out on the effect of garlic powder extract on fungi associated with Irish potato rot in Bokkos, Plateau State, Nigeria. Diseased Irish potatoes were randomly collected from three markets in the study location and fungal species isolated. Isolated fungal species were Fusarium culmorum, Fusarium oxysporum, and Pytophthora infestans. Frequency of occurrence for Fusarium culmorum, Fusarium oxysporum, and Pytophthora infestans was 10%, 34%, and 56%, respectively, using sabauraud dextrose agar, after incubation for 4-7 days. Treatment of Pytophthora infestans with garlic powder extract at concentrations of 0.5g/ml, 0.4g/ml, 0.3gml, 0.2g/ml and 0.1g/ml showed 100%, 92%, 68%, 32% and 10% inhibition zones, respectively. Fusarium culmorum showed 100%, 90%, 40%, 9% and 0% inhibition zones when treated with garlic powder extract at concentrations of 0.5g/ml, 0.4g/ml, 0.3gml, 0.2g/ml and 0.1g/ml, respectively. Garlic powder extract concentrations of 0.5g/ml, 0.4g/ml, 0.3gml, 0.2g/ml and 0.1g/ml showed 100%, 98%, 55%, 30%, 0% inhibition zones, respectively on Fusarium oxysporum. Hence, Restriction of the radial growth of the fungal colonies suggests a good antifungal effect of garlic extract. This can be integrated into the treatment of fungal diseases of Irish potato in Bokkos, Nigeria, as this will help to reduce the indiscriminate use of fungicides, especially in an environment with a struggling economy.

Keywords: fungal rot, garlic extract, inhibition zone, Irish potato

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Authors: Tamar Khardziani, Violeta Berikashvili, Mariam Rusitashvili, Eva Kachlishvili, Vladimir Elisashvili, Mikheil Asatiani

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Last years, the search for natural sources of novel and effective antifungal substances became a scientific and technological challenge. In the present research, thirty basidiomycetes isolated from various ecological niches of Georgia and belonging to different taxonomic groups were screened for their antifungal activities against pathogenic fungi such as Aspergillus, Fusarium, and Guignardia bidwellii. Among mushroom tested, several potential producers of antifungal substances have been revealed, such as Schizophyllum commune, Lentinula edodes, Ganoderma abietinum, Fomes fomentarius, Hericium erinaceus, and Trametes versicolor. For mushroom cultivation and expression of antifungal potential, submerged and solid-state fermentations of different plant raw materials were performed and various approaches and strategies have been exploited. Sch. commune appeared as a most promising producer of antifungal compounds. It was established that among different agro-industrial wastes, the presence of mandarin juice production waste in a nutrient medium, causing the significant increase of antifungal activity Sch. commune (growth inhibition: Aspergillus – 59 %, Fusarium – 55 %, G. bidwellii – 78 %, after 3, 2 and 4 days of cultivation, respectively). Besides this, Sch. commune demonstrate similar antifungal activities in the presence of glucose, glycerol, maltose, mannitol, and xylose, and growth inhibition of Fusarium ranged in 41 % - 49 % during 6 days of cultivation. Inhibition of Aspergillus growth inhibition varied in 27 % - 36 %, and inhibition of G. bidwellii was in the range 49 % - 61 %, respectively. Sch. commune under solid-state fermentation of mandarin peels at 13 days of cultivation demonstrates powerful growth inhibition of pathogenic fungi (growth inhibition: Aspergillus – 50 %, Fusarium – 61 %, G. bidwellii – 68 %, after 3, 4, and 4 days of cultivation, respectively) as well as at 20 days old mushroom (growth inhibition: Aspergillus – 41 %, Fusarium – 54 %, G. bidwellii – 66 %, after 3 days of cultivation). It was established that Sch. commune was effective as a producer of antifungal compounds in submerged as well as in solid-state fermentation. Finally, performed study confirms that the higher basidiomycetes possess antifungal potential, which strongly depends on the physiological factors of growth. Acknowledgments: The work was implemented with the financial support of fundamental science project FR-19-3719 by the Shota Rustaveli National Science Foundation of Georgia.

Keywords: antifungal potential, higher basidiomycetes, pathogenic fungi, submerged and solid-state fermentation

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Authors: Ana Tsokilauri

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The uppermost factor in the regulation of lignin-cellulose activity of decaying white rot or free rot are the substances serving as carbon and nitrogen nutrition of microorganisms and are considered as the most important factor of generative activity of white rot. The research object was Basidiomycete Fungi, peculiar and common in Georgia, and the separation of 10 of them as pure crops. The unidentified pure crops have tasted in order to be determined the potential of synthesis of lignin-degrading enzymes and the substrate of optimal lignocellulose growth. One of the most important aspects of the research conducted on Basidiomycetes was the use of specific lignocellulosic residues for selecting Fungi as a substrate of their growth. In order to increase lignocellulose with the help of substrate, crops were selected from the screening stage that showed good laccase activity. (Dusheti 1; Dusheti 10; Fshavi 5; Fshavi1; Fshavi 8; Fshavi 32; Manglisi 26; Sabaduri20; Dusheti 7; Sabaduri 1 ), Among the selected cultures, the crops with good laccase activity against the following substances, in particular: Dusheti 1- in this case, the rate of enzymatic activity on bran substrate was -105,6 u/ml, mandarin-96,4 u/ml. In case of corn - 102,9 u/ml. In case of Dusheti 7- the indicators were as follows: bananas-121,7 u/ml, mandarin-125,4 u/ml, corn - 117,1 u/ml. In the case of Sanaduri 32, the laccase activity was as follows: pomegranate- 101,2 u/ml. As a result of conducted experiments, the synthesis and activity rates of enzymes depending on plant substrates varied within a fairly wide range, which is still being under research.

Keywords: Lignocellulosic substrate, Basidiomycetes, white-rot basidiomycetes, Laccase

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Authors: Yui Okuno, Mariko Era, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Introduction: Fungus and mite are known as allergens that cause an allergic disease for example asthma bronchiale and allergic rhinitis. Cladosporium cladosporioides is one of the most often detected fungi in the indoor environment and causes pollution and deterioration. Dermatophagoides farinae is major mite allergens indoors. Therefore, the creation of antifungal agents with high safety and the antifungal effect is required. Fatty acid salts are known that have antibacterial activities. This report describes the effects of fatty acid salts against Cladosporium cladosporioides NBRC 30314 and Dermatophagoides farinae. Methods: Potassium salts of 9 fatty acids (C4:0, C6:0, C8:0, C10:0, C12:0, C14:0, C18:1, C18:2, C18:3) were prepared by mixing the fatty acid with the appropriate amount of KOH solution to a concentration of 175 mM and pH 10.5. The antifungal method, the spore suspension (3.0×104 spores/mL) was mixed with a sample of fatty acid potassium (final concentration of 175 mM). Samples were counted at 0, 10, 60, 180 min by plating (100 µL) on PDA. Fungal colonies were counted after incubation for 3 days at 30 °C. The MIC (minimum inhibitory concentration) against the fungi was determined by the two-fold dilution method. Each fatty acid salts were inoculated separately with 400 µL of C. cladosporioides at 3.0 × 104 spores/mL. The mixtures were incubated at the respective temperature for each organism for 10 min. The tubes were then contacted with the fungi incubated at 30 °C for 7 days and examined for growth of spores on PDA. The acaricidal method, twenty D. farinae adult females were used and each adult was covered completely with 2 µL fatty acid potassium for 1 min. The adults were then dried with filter paper. The filter paper was folded and fixed by two clips and kept at 25 °C and 64 % RH. Mortalities were determained 48 h after treatment under the microscope. D. farina was considered to be dead if appendages did not move when prodded with a pin. Results and Conclusions: The results show that C8K, C10K, C12K, C14K was effective to decrease survival rate (4 log unit) of the fatty acids potassium incubated time for 10 min against C. cladosporioides. C18:3K was effective to decrease 4 log unit of the fatty acids potassium incubated time for 60 min. Especially, C12K was the highest antifungal activity and the MIC of C12K was 0.7 mM. On the other hand, the fatty acids potassium showed no acaricidal effects against D. farinae. The activity of D. farinae was not adversely affected after 48 hours. These results indicate that C12K has high antifungal activity against C. cladosporioides and suggest the fatty acid potassium will be used as an antifungal agent.

Keywords: fatty acid salts, antifungal effects, acaricidal effects, Cladosporium cladosporioides, Dermatophagoides farinae

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Authors: Lena Payati, Maria Kazou, Effie Tsakalidou

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Table olives are one of the most popular fermented vegetables worldwide, which along with olive oil, have a crucial role in the world economy. They are highly appreciated by the consumers for their characteristic taste and pleasant aromas, while several health and nutritional benefits have been reported as well. Until recently, microbial biogeography, i.e., the study of microbial diversity over time and space, has been mainly associated with wine. However, nowadays, the term 'terroir' has been extended to other crops and food products so as to link the geographical origin and environmental conditions to quality aspects of fermented foods. Taking the above into consideration, the present study focuses on the microbial fingerprinting of the most important olive varieties of Greece with the state-of-the-art amplicon-based metagenomics analysis. Towards this, in 2019, 61 samples from 38 different olive varieties were collected at the final stage of ripening from 13 well spread geographical regions in Greece. For the metagenomics analysis, total DNA was extracted from the olive samples, and the 16S rRNA gene and ITS DNA region were sequenced and analyzed using bioinformatics tools for the identification of bacterial and yeasts/fungal diversity, respectively. Furthermore, principal component analysis (PCA) was also performed for data clustering based on the average microbial composition of all samples from each region of origin. According to the composition, results obtained, when samples were analyzed separately, the majority of both bacteria (such as Pantoea, Enterobacter, Roserbergiella, and Pseudomonas) and yeasts/fungi (such as Aureobasidium, Debaromyces, Candida, and Cladosporium) genera identified were found in all 61 samples. Even though interesting differences were observed at the relative abundance level of the identified genera, the bacterial genus Pantoea and the yeast/fungi genus Aureobasidium were the dominant ones in 35 and 40 samples, respectively. Of note, olive samples collected from the same region had similar fingerprint (genera identified and relative abundance level) regardless of the variety, indicating a potential association between the relative abundance of certain taxa and the geographical region. When samples were grouped by region of origin, distinct bacterial profiles per region were observed, which was also evident from the PCA analysis. This was not the case for the yeast/fungi profiles since 10 out of the 13 regions were grouped together mainly due to the dominance of the genus Aureobasidium. A second cluster was formed for the islands Crete and Rhodes, both of which are located in the Southeast Aegean Sea. These two regions clustered together mainly due to the identification of the genus Toxicocladosporium in relatively high abundances. Finally, the Agrinio region was separated from the others as it showed a completely different microbial fingerprinting. However, due to the limited number of olive samples from some regions, a subsequent PCA analysis with more samples from these regions is expected to yield in a more clear clustering. The present study is part of a bigger project, the first of its kind in Greece, with the ultimate goal to analyze a larger set of olive samples of different varieties and from different regions in Greece in order to have a reliable olives’ microbial biogeography.

Keywords: amplicon-based metagenomics analysis, bacteria, microbial biogeography, olive microbiota, yeasts/fungi

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Authors: Ayda Maadani Mallak, Amir lakzian, Elham Khodaverdi, Gholam Hossein Haghnia

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The use of pharmaceuticals and personal care products such as antibiotics and antibacterials has been increased in recent years. Since the major part of consumed compounds remains unchanged in the wastewater treatment plant, they will easily find their way into the human food chain following the land use of sewage sludge (SS). Biological treatment of SS is one the most effective methods for expunging contaminants. White rot fungi, due to their ligninolytic enzymes, are extensively used to degrade organic compounds. Among all three different morphological forms and growth patterns of filamentous fungi (mycelia, clumps, and pellets), fungal pellet formation has been the subject of interest in industrial bioprocesses. Therefore this study was aimed to investigate the uptake of tetracycline (TC) and triclosan (TCS) by radish plant (Raphanus sativus) from soil amended with untreated and pretreated SS by P. ostreatus fungal pellets under greenhouse conditions. The experimental soil was amended with 1) Contaminated SS with TC at a concentration of 100 mgkg-1 and pretreated by fungal pellets, 2) Contaminated SS with TC at 100 mgkg-1 and untreated with fungal pellets, 3) Contaminated SS with TCS at a concentration of 50 mgkg-1 and pretreated by fungal pellets, 4) contaminated SS with TCS at 50 mgkg-1 and untreated with fungal pellets. An uncontaminated and untreated SS-amended soil also was considered as control treatment. An AB SCIEX 3200 QTRAP LC-MS/MS system was used in order to analyze the concentration of TC and TCS in plant tissues and soil medium. Results of this study revealed that the presence of TC and TCS in SS-amended soil decreased the radish biomass significantly. The reduction effect of TCS on dry biomass of shoot and root was 39 and 45% compared to controls, whereas for TC, the reduction percentage for shoot and root was 27 and 40.6%, respectively. However, fungal treatment of SS by P. ostreatus pellets reduced the negative effect of both compounds on plant biomass remarkably, as no significant difference was observed compared to control treatments. Pretreatment of SS with P. ostreatus also caused a significant reduction in translocation factor (concentration in shoot/root), especially for TC compound up to 32.3%, whereas this reduction for TCS was less (8%) compared to untreated SS. Generally, the results of this study confirmed the positive effect of using fungal pellets in SS amendment to decrease TC and TCS uptake by radish plants. In conclusion, P. ostreatus fungal pellets might provide future insights into bioaugmentation to remove antibiotics from environmental matrices.

Keywords: antibiotic, fungal pellet, sewage sludge, white-rot fungi

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Authors: Stavros Palavouzis, Alexandra Triantafyllopoulou, Aliki Tzima, Epaminondas Paplomatas

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Mating-type genes are present in most filamentous fungi, even though teleomorphs for all species have not been recorded. Our study tries to explore the effect of different growth conditions on the expression of MAT genes in Neofusicoccum mediterraneum. As such, selected isolates were grown in potato dextrose broth or in water agar supplemented with pine needles under a 12 h photoperiod, as well as in constant darkness. Mycelia and spores were collected at different time points, and RNA extraction was performed, with the extracted product being used for cDNA synthesis. New primers for MAT gene expression were designed while qPCR results are underway. The second part of the study involved the isolation and cloning in a selected pGEM-T vector of the Botryosphaeria dothidea MAT1 1 1 and MAT1 2 1 mating genes, including flanking regions. As a next step, the genes were amplified using newly designed primers with engineered restriction sites. Amplicons were excised and subsequently sub-cloned in appropriate binary vectors. The constructs were afterward inserted into Agrobacterium tumefaciens and utilized for Agrobacterium-mediated transformation (ATMT) of Neofusicoccum mediterraneum. At the same time, the transformation of a Verticillium dahliae tomato race 1 strain (70V) was performed as a control. While the procedure was successful in regards to V. dahliae, transformed strains of N. mediterraneum could not be obtained. At present, a new transformation protocol, which utilizes a combination of protoplast and Agro transformation, is being evaluated.

Keywords: anamorph, heterothallism, perithecia, pycnidia, sexual stage

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Authors: Sarunpron Khruengsai, Patcharee Pripdeevech

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This study aimed to investigate the fumigation activities of the volatile compounds produced by Trichoderma spp. against Fusarium oxysporum and F. proliferatum fungi that cause significant rot in fresh chilies. Two Trichoderma spp. were isolated from the leaves of Schefflera leucantha grown in Thailand and later identified as T. afroharzianum MFLUCC19-0090 and T. afroharzianum MFLUCC19-0091. Both in vitro and in vivo dual culture volatile assays were used to study the effects of the produced volatile compounds on mycelial growth. In vitro results showed that the volatile compounds produced by T. afroharzianum MFLUCC19-0090 significantly inhibited the growth of F. oxysporum, while the volatile compounds produced by T. afroharzianum MFLUCC19-0091 significantly inhibited the growth of F. proliferatum. The effectiveness of Trichoderma-derived volatile compounds in inhibiting the mycelial growth of the selected pathogens in the inoculated, fresh chili samples was further demonstrated in vivo. The volatile profiles of both Trichoderma spp. were characterized using gas chromatography-mass spectrometry. Seventy-three volatile compounds were detected from both strains. Among the major volatile compounds detected, phenyl ethyl alcohol was found to possess the strongest antifungal activity against both pathogens. The results support the possibility of using volatile compounds produced by T. afroharzianum MFLUCC19-0090 and T. afroharzianum MFLUCC19-0091 as alternative fumigants for preventing Fusarium rot of fresh chilies during the post-harvest period.

Keywords: antifungal activity, biocontrol, endophytic fungi, post-harvest

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Authors: Yaara Oppenheimer-Shaanan, Nimrod Nachmias, Marina Campos Rocha, Neta Schlezinger, Noam Dotan, Asaf Levy

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Fusarium oxysporum, a fungus that attacks a broad range of plants and can cause infections in humans, operates across different kingdoms. This pathogen encounters varied conditions, such as temperature, pH, and nutrient availability, in plant and human hosts. The Fusarium oxysporum species complex, pervasive in soils globally, can affect numerous plants, including key crops like tomatoes and bananas. Controlling Fusarium infections can involve biocontrol agents that hinder the growth of harmful strains. Our research developed a computational method to identify toxin domains within a vast number of microbial genomes, leading to the discovery of nine distinct toxins capable of killing bacteria and fungi, including Fusarium. These toxins appear to function as enzymes, causing significant damage to cellular structures, membranes and DNA. We explored biological control using bacteria that produce polymorphic toxins, finding that certain bacteria, non-pathogenic to plants, offer a safe biological alternative for Fusarium management, as they did not harm macrophage cells or C. elegans. Additionally, we elucidated the 3D structures of two toxins with their protective immunity proteins, revealing their function as unique DNases. These potent toxins are likely instrumental in microbial competition within plant ecosystems and could serve as biocontrol agents to mitigate Fusarium wilt and related diseases.

Keywords: microbial toxins, antifungal, Fusarium oxysporum, bacterial-fungal intreactions

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Authors: Mun Yee Chan, Sin Ming Goh, Lisa Gaik Ai Ong

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Approximately 10,000 different types of dyes and pigments are being used in various industrial applications yearly, which include the textile and printing industries. However, these dyes are difficult to degrade naturally once they enter the aquatic system. Their high persistency in natural environment poses a potential health hazard to all form of life. Hence, there is a need for alternative dye removal strategy in the environment via bioremediation. In this study, fungi laccase is investigated via commercial agar dyes plates and submerged fermentation to explore the application of fungi laccase in textile dye wastewater treatment. Two locally isolated basidiomycetes were screened for laccase activity using media added with commercial dyes such as 2, 2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), guaiacol and Remazol Brillant Blue R (RBBR). Isolate TBB3 (1.70±0.06) and EL2 (1.78±0.08) gave the highest results for ABTS plates with the appearance of greenish halo on around the isolates. Submerged fermentation performed on Isolate TBB3 with the productivity 3.9067 U/ml/day, whereas the laccase activity for Isolate EL2 was much lower (0.2097 U/ml/day). As isolate TBB3 showed higher laccase production, it was subjected to molecular characterization by DNA isolation, PCR amplification and sequencing of ITS region of nuclear ribosomal DNA. After being compared with other sequences in National Center for Biotechnology Information (NCBI database), isolate TBB3 is probably from species Trametes hirsutei. Further research work can be performed on this isolate by upscale the production of laccase in order to meet the demands of the requirement for higher enzyme titer for the bioremediation of textile dyes.

Keywords: bioremediation, dyes, fermentation, laccase

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Authors: Vanda Ferreira Ribeiro, Daiane Tomacheski, Douglas Naue Simões, Michele Pitto, Ruth Marlene Campomanes Santana

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Indoor environments, such as car cabins and public transportation vehicles are places where users are subject to air quality. Microorganisms (bacteria, fungi, yeasts) enter these environments through windows, ventilation systems and may use the organic particles present as a growth substrate. In addition, atmospheric pollutants can act as potential carbon and nitrogen sources for some microorganisms. Compounds base SEBS copolymers, poly(styrene-b-(ethylene-co-butylene)-b-styrene, are a class of thermoplastic elastomers (TPEs), fully recyclable and largely used in automotive parts. Metals, such as cooper and silver, have biocidal activities and the production of the SEBS compounds by melting blending with these agents can be a good option for producing compounds for use in plastic parts of ventilation systems and automotive air-conditioning, in order to minimize the problems caused by growth of pathogenic microorganisms. In this sense, the aim of this work was to evaluate the effect of copper microparticles as antimicrobial agent in compositions based on SEBS/PP/oil/calcite. Copper microparticles were used in weight proportion of 0%, 1%, 2% and 4%. The compounds were prepared using a co-rotating double screw extruder (L/D ratio of 40/1 and 16 mm screw diameter). The processing parameters were 300 rpm of screw rotation rate, with a temperature profile between 150 to 190°C. SEBS based TPE compounds were injection molded. The compounds emission were characterized by gravimetric fogging test. Compounds were characterized by physical (density and staining by contact), mechanical (hardness and tension properties) and rheological properties (melt volume rate – MVR). Antibacterial properties were evaluated against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) strains. To avaluate the abilities toward the fungi have been chosen Aspergillus niger (A. niger), Candida albicans (C. albicans), Cladosporium cladosporioides (C. cladosporioides) and Penicillium chrysogenum (P. chrysogenum). The results of biological tests showed a reduction on bacteria in up to 88% in E.coli and up to 93% in S. aureus. The tests with fungi showed no conclusive results because the sample without copper also demonstrated inhibition of the development of these microorganisms. The copper addition did not cause significant variations in mechanical properties, in the MVR and the emission behavior of the compounds. The density increases with the increment of copper in compounds.

Keywords: air conditioner, antimicrobial, cooper, SEBS

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Authors: Carlos E. Rodríguez-Rodríguez, Karla Ruiz-Hidalgo, Kattia Madrigal-Zúñiga, Juan Salvador Chin-Pampillo, Mario Masís-Mora, Elizabeth Carazo-Rojas

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One of the main causes of contamination linked to agricultural activities is the spillage and disposal of pesticides, especially during the loading, mixing or cleaning of agricultural spraying equipment. One improvement in the handling of pesticides is the use of biopurification systems (BPS), simple and cheap degradation devices where the pesticides are biologically degraded at accelerated rates. The biologically active core of BPS is the biomixture, which is constituted by soil pre-exposed to the target pesticide, a lignocellulosic substrate to promote the activity of ligninolitic fungi and a humic component (peat or compost), mixed at a volumetric proportion of 50:25:25. Considering the known ability of lignocellulosic fungi to degrade a wide range of organic pollutants, and the high amount of lignocellulosic waste used in biomixture preparation, the bioaugmentation of biomixtures with these fungi represents an interesting approach for improving biomixtures. The present work aimed at evaluating the effect of the bioaugmentation of rice husk based biomixtures with the fungus Trametes versicolor in the removal of the insectice/nematicide carbofuran (CFN) and to optimize the composition of the biomixture to obtain the best performance in terms of CFN removal and mineralization, reduction in formation of transformation products and decrease in residual toxicity of the matrix. The evaluation of several lignocellulosic residues (rice husk, wood chips, coconut fiber, sugarcane bagasse or newspaper print) revealed the best colonization by T. versicolor in rice husk. Pre-colonized rice husk was then used in the bioaugmentation of biomixtures also containing soil pre-exposed to CFN and either peat (GTS biomixture) or compost (GCS biomixture). After spiking with 10 mg/kg CBF, the efficiency of the biomixture was evaluated through a multi-component approach that included: monitoring of CBF removal and production of CBF transformation products, mineralization of radioisotopically labeled carbofuran (14C-CBF) and changes in the toxicity of the matrix after the treatment (Daphnia magna acute immobilization test). Estimated half-lives of CBF in the biomixtures were 3.4 d and 8.1 d in GTS and GCS, respectively. The transformation products 3-hydroxycarbofuran and 3-ketocarbofuran were detected at the moment of CFN application, however their concentration continuously disappeared. Mineralization of 14C-CFN was also faster in GTS than GCS. The toxicological evaluation showed a complete toxicity removal in the biomixtures after 48 d of treatment. The composition of the GCS biomixture was optimized using a central composite design and response surface methodology. The design variables were the volumetric content of fungally pre-colonized rice husk and the volumetric ratio compost/soil. According to the response models, maximization of CFN removal and mineralization rate, and minimization in the accumulation of transformation products were obtained with an optimized biomixture of composition 30:43:27 (pre-colonized rice husk:compost:soil), which differs from the 50:25:25 composition commonly employed in BPS. Results suggest that fungal bioaugmentation may enhance the performance of biomixtures in CFN removal. Optimization reveals the importance of assessing new biomixture formulations in order to maximize their performance.

Keywords: bioaugmentation, biopurification systems, degradation, fungi, pesticides, toxicity

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Authors: Zhazira Shemsheyeva, Zhanara Suleimenova, Olga Shemshura, Gulnaz Mombekova, Zhanar Rakhmetova

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Bacteria that colonize plant roots and promote plant growth are referred to as plant growth-promoting rhizobacteria (PGPR). They not only provide nutrients to the plants (direct plant growth promotion) and protect plants against the phytopathogens (indirect plant growth promotion) but also increase the soil fertility. Indirectly PGPRs improve the plant growth by becoming a biocontrol agent for a fungal pathogen. The antifungal activities of the PGPrhizobacteria were assayed against different species of phytopathogenic fungi such as Fusarium tricinctum, Fusarium oxysporum, Sclerotiniasclerotiorum, and Botrytis cinerea. Pseudomonas putidaSM-1, Azotobacter sp., and Bacillus thuringiensis AKS/16 strains have been used in experimental tests on growth inhibition of phytopathogenic fungi infecting Kidney beans. Agar well diffusion method was used in this study. Diameters of the zones of inhibition were measured in millimeters. It was found that Bacillus thuringiensis AKS/16 strain showed the lowest antifungal activity against all fungal pathogens tested. Zones of inhibition were 15-18 mm. In contrast, Pseudomonas putida SM-1 exhibited good antifungal activity against Fusarium oxysporum and Fusarium tricinctum by producing 29-30 mm clear zones of inhibition. The moderate inhibitory effect was shown by Azotobacter sp. against all fungal pathogens tested with zones of inhibition from24 to 26 mm. In summary, Pseudomonas putida SM-1 strain demonstrated the potential of controlling root rot diseases in kidney beans.

Keywords: PGPR, pseudomonas putida, kindey beans, antifungal activity

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Authors: Marzieh Fotovvat, Farzaneh Najafi, Ramazan Ali Khavari-Nejad, Daryush Talei, Farhad Rejali

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Salvia leriifolia Benth. is one of the valuable and perennial medicinal plants of the Lamiaceae family, geographically growing in the south and tropical regions of Khorassan and Semnan provinces in Iran. In recent years, several medicinal properties such as antimicrobial, antifungal, anti-diabetic, analgesic, and anti-inflammatory effects have been reported from this plant. The use of elicitors such as chitosan and Arbuscular mycorrhizal fungi (AMF) symbiosis are the main methods for increasing the production of secondary metabolites, growth, and physiological factors in plants. The main aim of this study was to investigate the effects of foliar spraying applications by chitosan and/or the contribution of AMF (Glomus interaradices) on some growth factors and chlorophyll content of S. leriifolia under glasshouse conditions. The sterilized seeds were germinated by placing them into a cocopeat. After one month, seedlings that were in the 2-4 leaf stage were transferred to plastic pots (garden soil and pumice at 2:1) with or without mycorrhizal fungi. Chitosan (0, 50, 100, 200, and 400 mg L-1) was sprayed four times in the fourth month of the vegetative period. The results showed that fresh leaf weight, fresh root weight, root height, and chlorophyll content could change in the plant treated with chitosan and AMF symbiosis. So that the highest chlorophyll content and fresh weight of roots and leaves were observed in the interaction of chitosan and G. interaradices. In general, by optimizing the chitosan concentration and the use of appropriate AMF symbiosis, it is possible to improve the growth and quality of the medicinal plant S. leriifolia.

Keywords: chitosan, chlorophyll, growth factors, mycorrhiza

Procedia PDF Downloads 83

Authors: Aya Tanaka, Mariko Era, Shiho Sakai, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives: Aspergillus niger and Aspergillus oryzae are used as koji fungi in the spot of the brewing. Since koji-muro (room for making koji) was a low level of airtightness, microbial contamination has long been a concern to the alcoholic beverage production. Therefore, we focused on the fatty acid salt which is the main component of soap. Fatty acid salts have been reported to show some antibacterial and antifungal activity. So this study examined antimicrobial activities against Aspergillus and Bacillus spp. This study aimed to find the effectiveness of the fatty acid salt in koji-muro as antimicrobial agents. Materials & Methods: A. niger NBRC 31628, A. oryzae NBRC 5238, A. oryzae (Akita Konno store) and Bacillus subtilis NBRC 3335 were chosen as tested. Nine fatty acid salts including potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K) and linolenate (C18:3K) at 350 mM and pH 10.5 were used as antimicrobial activity. FASs and spore suspension were prepared in plastic tubes. The spore suspension of each fungus (3.0×104 spores/mL) or the bacterial suspension (3.0×105 CFU/mL) was mixed with each of the fatty acid salts (final concentration of 175 mM). The mixtures were incubated at 25 ℃. Samples were counted at 0, 10, 60, and 180 min by plating (100 µL) on potato dextrose agar. Fungal and bacterial colonies were counted after incubation for 1 or 2 days at 30 ℃. The MIC (minimum inhibitory concentration) is defined as the lowest concentration of drug sufficient for inhibiting visible growth of spore after 10 min of incubation. MICs against fungi and bacteria were determined using the two-fold dilution method. Each fatty acid salt was separately inoculated with 400 µL of Aspergillus spp. or B. subtilis NBRC 3335 at 3.0 × 104 spores/mL or 3.0 × 105 CFU/mL. Results: No obvious change was observed in tested fatty acid salts against A. niger and A. oryzae. However, C12K was the antibacterial effect of 5 log-unit incubated time for 10 min against B. subtilis. Thus, C12K suppressed 99.999 % of bacterial growth. Besides, C10K was the antibacterial effect of 5 log-unit incubated time for 180 min against B. subtilis. C18:1K, C18:2K and C18:3K was the antibacterial effect of 5 log-unit incubated time for 10 min against B. subtilis. However, compared to saturated fatty acid salts to unsaturated fatty acid salts, saturated fatty acid salts are lower cost. These results suggest C12K has potential in the field of koji-muro. It is necessary to evaluate the antimicrobial activity against other fungi and bacteria, in the future.

Keywords: Aspergillus, antimicrobial, fatty acid salts, koji-muro

Procedia PDF Downloads 555

Authors: Salama A. Ouf, Amera A. El-Adly, Abdelaleam H. Mohamed

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Dermatophytosis is the infection of keratinized tissues such as hair, nail and the stratum corneum of the skin by dermatophytic fungi. Infection is generally cutaneous and restricted to the non-living cornified layers because of the inability of the fungi to penetrate the deeper tissues or organs of immunocompetent hosts. In Saudi Arabia, Onychomycosis is the most frequent infection (40.3%), followed by tinea capitis (21.9%), tinea pedis (16%), tinea cruris (15.1%), and tinea corporis (6.7%). Several azole compounds have been tried to control dermatophytic infection, however, the azole-containing medicines may interfere with the activity of hepatic microsomal enzymes, sex and thyroid hormones, and testosterone biosynthesis. In this research, antibody-conjugated nanoparticles stimulated by cold plasma and laser were evaluated in vitro against some dermatophytes isolated from the common types of tinea. Different types of nanomaterials were tested but silver nanoparticles (AgNPs) were proved to be most effective against the dermatophytes under test. The use of cold plasma coupled with antibody-conjugated nano-particles has severe impact on dermatophytes where the inhibition of growth, spore germination keratinase activity was more than 88% in the case of Trichophyton rubrum, T. violaceum, Microsprum canis and M. gypseum. Complete inhibition of growth for all dermatophytes was brought about by the interaction of conjugated nanoparticles, with cold plasma and laser treatment. The in vivo test with inoculated guinea pigs achieved promising results where the recovery from the infection reached 95% in the case of M. canis –inoculated pigs treated with AgNPs pretreated with cold plasma and laser.

Keywords: cold plasma, dermatophytes, laser, silver nanoparticles

Procedia PDF Downloads 367

Authors: Hyuk Sung Yoon, In-Lee Choi, Mohammad Zahirul Islam, Jun Pill Baek, Ho-Min Kang

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The strawberry ‘Maehyang’ cultivated in South Korea has been increased to export to Southeast Asia. The degradation of quality often occurs in strawberries during short export period. Botrytis cinerea has been known to cause major damage to the export strawberries and the disease was caused during shipping and distribution. This study was conducted to find out the sterilized effect of chlorine dioxide(ClO2) gas and high concentration of CO2 gas injection for ‘Maehyang’ strawberry and it was packaged with oxygen transmission rate (OTR) films. The strawberry was harvested at 80% color changed stage and packaged with OTR film and perforated film (control). The treatments were a MAP used by with 20,000 cc·m-2·day·atm OTR film and gas injection in packages. The gas type of ClO2 and CO2 were injected into OTR film packages, and treatments were 6 mg/L ClO2, 15% CO2, and they were combined. The treated strawberries were stored at 3℃ for 30 days. Fresh weight loss rate was less than 1% in all OTR film packages but it was more than 15% in a perforated film treatment that showed severe deterioration of visual quality during storage. Carbon dioxide concentration within a package showed approximately 15% of the maximum CO2 concentration in all treatments except control until the 21st day, it was the tolerated range of maximum CO2 concentration of strawberry in recommended CA or MA conditions. But, it increased to almost 50% on the 30th day. Oxygen concentration showed a decrease down to approximately 0% in all treatments except control for 25 days. Ethylene concentration was shown to be steady until the 17th day, but it quickly increased on the 17th day and dropped down on the final storage day (30th day). All treatments did not show any significant differences in gas treatments. Firmness increased in CO2 (15%) and ClO2 (6mg/L) + CO2 (15%) treatments during storage. It might be the effect of high concentration CO2 known by reducing decay and cell wall degradation. The soluble solid decreased in all treatments during storage. These results were caused to use up the sugar by the increase of respiration during storage. The titratable acidity showed a similarity in all treatments. Incidence of fungi was 0% in CO2 (15%) and ClO2 (6mg/L)+ CO2 (15%), but was more than 20% in a perforated film treatment. Consequently, The result indicates that Chloride Dioxide(ClO2) and high concentration of CO2 inhibited fungi growth. Due to the fact that fresh weight loss rate and incidence of fungi were lower, the ClO2(6mg/L)+ CO2(15%) prove to be most efficient in sterilization. These results suggest that Chloride Dioxide (ClO2) and high concentration of CO2 gas injection treatments were an effective decontamination technique for improving the safety of strawberries.

Keywords: chloride dioxide, high concentration of CO2, modified atmosphere condition, oxygen transmission rate films

Procedia PDF Downloads 339

Authors: L. Kutateldze, T. Urushadze, R. Khvedelidze, N. Zakariashvili, I. Khokhashvili, T. Sadunishvili

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Microbial cellulase/xylanase have shown their potential application in various industries including pulp and paper, textile, laundry, biofuel production, food and feed industry, brewing, and agriculture. Extremophilic micromycetes and their enzymes that are resistant to critical values of temperature and pH, and retaining enzyme activity for a long time are of great industrial interest. Among strains of microscopic fungi from the collection of S. Durmishidze Institute of Biochemistry and Biotechnology, strains isolated from different ecological niches of Southern Caucasus-active producers of cellulase/xylanase have been selected by means of screening under deep cultivation conditions. Extremophilic micromycetes and their enzymes that are resistant to critical values of temperature and pH, and retaining enzyme activity for a long time are of great industrial interest. Among strains of microscopic fungi from the collection of S. Durmishidze Institute of Biochemistry and Biotechnology, strains isolated from different ecological niches of Southern Caucasus-active producers of cellulase/xylanase have been selected by means of screening under deep cultivation conditions. Representatives of the genera Aspergillus, Penicillium and Trichoderma are outstanding by relatively high activities of these enzymes. Among the producers were revealed thermophilic strains, representatives of the genus Aspergillus-Aspergillus terreus, Aspergillus versicolor, Aspergillus wentii, also strains of Sporotrichum pulverulentum and Chaetomium thermophile. As a result of optimization of cultivation media and conditions, activities of enzymes produced by the strains have been increased by 4 -189 %. Two strains, active producers of cellulase/xylanase – Penicillium canescence E2 (mesophile) and Aspergillus versicolor Z17 (thermophile) were chosen for further studies. Cellulase/xylanase enzyme preparations from two different genera of microscopic fungi Penicillium canescence E2 and Aspergillus versicolor Z 17 were obtained with activities 220 U/g /1200 U/g and 125 U/g /940 U/g, correspondingly. Main technical characteristics were as follows: the highest enzyme activities were obtained for mesophilic strain Penicillium canescence E2 at 45-500C, while almost the same enzyme activities were fixed for the thermophilic strain Aspergillus versicolor Z 17 at temperature 60-65°C, exceeding the temperature optimum of the mesophile by 150C. Optimum pH of action of the studied cellulase/xylanases from mesophileic and thermophilic strains were similar and equaled to 4.5-5.0 It has been shown that cellulase/xylanase technical preparations from selected strains of Penicillium canescence E2 and Aspergillus versicolor Z17 hydrolyzed cellulose of untreated wheat straw to reducible sugars by 46-52%, and to glucose by 22-27%. However the thermophilic enzyme preparations from the thermophilic A.versicolor strains conducted the process at 600C higher by 100C as compared to mesophlic analogue. Rate of hydrolyses of the pretreated substrate by the same enzyme preparations to reducible sugars and glucose conducted at optimum for their action 60 and 500C was 52-61% and 29-33%, correspondingly. Thus, maximum yield of glucose and reducible sugars form untreated and pretreated wheat straw was achieved at higher temperature (600C) by enzyme preparations from thermophilic strain, which gives advantage for their industrial application.

Keywords: cellulase/xylanase, cellulose hydrolysis, microscopic fungi, thermophilic strain

Procedia PDF Downloads 288

Authors: Ju Young Kim, Mohammad Zahirul Islam, Mahmuda Akter Mele, Su Jeong Han, Hyuk Sung Yoon, In-Lee Choi, Ho-Min Kang

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Strawberry and tomato fruits were harvested at the red ripens maturity stage in the Republic of Korea. The fruits were dipped in fungi solution and afterwards were sterilized with sodium hypochlorite (NaOCl) and chlorine dioxide (ClO2) gas. Some fruits were dipped in 150μL/L NaOCl solution for 10 minutes, and others were treated with 5μL/L ClO2 gas for 12 hours and packed with 20,000 cc OTR (oxygen transmission rate) film, the rest were packed in 10,000 cc OTR film inserted with 5μL/L ClO2 gas. 5μL/L ClO2 gas insert treatment showed the lowest carbon dioxide and ethylene, and the highest oxygen concentration was on the final storage day (15th day) in both strawberry and tomato fruits. Tomato fruits showed the lowest fresh weight loss in 5μL/L ClO2 gas insert treatment. The visual quality as well as shelf life showed the highest in 5μL/L ClO2 gas insert treatment of both strawberry and tomato fruits. In addition, the fungal incidence of strawberry and tomato fruits were the most suppressed in 5μL/L ClO2 gas insert treatment. 5μL/L ClO2 gas insert treatment showed higher firmness and soluble solids in both strawberry and tomato fruits. So, 5μL/L ClO2 gas insert treatment may be useful to prevent the fungal incidence as well as retaining the postharvest quality, and increase the shelf life of strawberry and tomato fruits for long term storage. This study was supported by Export Promotion Technology Development Program (314027-03), IPET, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea.

Keywords: chlorine dioxide, ethylene, fungi, sodium hypochlorite

Procedia PDF Downloads 366

Authors: Digby Wrede, Stephen Gray, Syed Hussainy

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Microalgae are extremely useful organisms but notoriously hard to harvest. The use of fungal pellets has been found to be an efficient way to flocculate numerous species of algae. However, only the flocculation of single species of algae has been investigated. Algae are generally found in complex communities in the environment comprising of numerous species of algae ranging from simple single cell algae such as Chlorella to more complex or communal algae such as Dictyosphaerium. This study investigated the flocculation capabilities of Aspergillus oryzae to flocculate four species of algae; Chlorella vulgaris, Scenedesmus quadricauda, Scenedesmus acuminatus and Dictyosphaerium sp., and the algal communities in four different types of domestic effluent from a lagoon-based treatment plant; primary effluent, secondary effluent and the high rate algal pond effluent at a natural and at a lowered pH level. Spectrophotometry was used to measure the changes in algal population. C. vulgaris, S. acuminatus and S. quadricauda, had over 90% reduction of algal in suspension after 24 hours. Dictyosphaerium sp. showed a little to no removal after 24 hours. The primary, secondary, and natural pH level HRAP had roughly a 50% removal after 24 hours, the HRAP which was grown at a lower pH level had over a 90% removal after 24 hours. pH has been shown previously to affect fungal flocculation. Fungal and algae pellets have been shown to be able to treat wastewater and can be converted to biofuels in a very similar method to how algae are currently converted. The mixture of both fungi and algae has also been shown to provide a higher yield of oils then separately and are able to more efficiently treat wastewater then algae or fungi by themselves.

Keywords: algae harvesting, Aspergillus oryzae, fungal flocculation, wastewater treatment

Procedia PDF Downloads 164

Authors: Baba John, Abdullahi Mohammed

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The aim of this research is to compare the analysis of biodegradation on polythene and plastics buried in fadama soil amended with Organic and Inorganic fertilizer. Physico- chemical properties of the samples were determined. Bacteria and Fungi implicated in the biodegradation were identified and enumerated. Physico- chemical properties before the analysis indicated pH range of the samples from 4.28 – 5.80 , While the percentage Organic carbon and Organic matter was highest in cow dung samples with 3.89% and 6.69% respectively. The total Nitrogen percentage was recorded to be highest in Chicken dropping (0.68), while the availability of Phosphorus (P), Sodium (Na), Pottasium (K), and Magnessium (mg) was recorded to be highest in F – soil (Control), with values to be 37ppm, 1.63 Cmolkg-1, 0.35 Cmolkg-1 and 1.18 Cmolkg-1 respectively, except for calcium which was recorded to be highest in Cow dung (5.80 Cmolkg-1). However, physico – chemical properties of the samples after analysis indicated pH range of 4.6 – 5.80, Percentage Organic carbon and Organic matter was highest in Fadama soil mixed with fertilizer, having 0.7% and 1.2% respectively. Total Percentage Nitrogen content was found to be highest (0.56) in Fadama soil mixed with poultry dropping. Availability of Sodium (Na), Pottasium (K), and Calcium (Ca) was recorded to be highest in Fadama Soil mixed with Cow dung with values to be 0.64 Cmolkg-1, 2.07 Cmolkg-1 and 3.36 Cmolkg-1 respectively. The percentage weight loss of polythene and plastic bags after nine months in fadama soil mixed with poultry dropping was 11.9% for polythene and 6.0% for plastics. Weight loss in fadama soil mixed with cow dung was 18.1% for polythene and 4.7% for plastics. Weight loss of polythene and plastic in fadama soil mixed with fertilizer (NPK) was 7.4% for polythene and 3.3% for plastics. While, the percentage weight loss of polythene and plastics after nine months of burial in fadama soil (control) was 3.5% and 0.0% respectively. The bacteria species isolated from Fadama soil, organic and inorganic fertilizers before amendments include: S. aureus, Micrococcus sp, Streptococcus. pyogenes, Psuedomonas aeruginosa Bacillus subtilis and Bacillus cereus. The fungi species include: Aspergillus niger, Aspergillus fumigatus, Aspergillus flavus, Fusarium sp, Mucor sp Penicillium sp and Candida sp. The bacteria species isolated and characterized after nine months of seeding include: S. aureus, Micrococcus sp, S. pyogenes, P. aeruginosa and B. subtilis. The fungi species are: A. niger A. flavus, A. fumigatus, Mucor sp, Penicillium sp and Fusarium sp. The result of this study indicated that plastic materials can be degraded in the fadama soil irrespective of whether the soil is amended or not. The Period of composting also has a significant impact on the rate at which polythene and plastics are degraded.

Keywords: Fadama, fertilizer, plastic and polythene, biodegradation

Procedia PDF Downloads 544

Authors: Maria P. Geneva, Ira V. Stancheva, Marieta G. Hristozkova, Roumiana D. Vasilevska-Ivanova, Mariana T. Sichanova, Janet R. Mincheva

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Hyssopus officinalis L., Lamiaceae, commonly called hyssop, is an aromatic, semi-evergreen, woody-based, shrubby perennial plant. Hyssop is a good expectorant and antiviral herb commonly used to treat respiratory conditions such as influenza, sinus infections, colds, and bronchitis. Most of its medicinal properties are attributed to the essential oil of hyssop. The study was conducted to evaluate the influence of inoculation with arbuscular mycorrhizal fungi of in vitro propagated hyssop plants on the: activities of antioxidant enzymes superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase; accumulation of non-enzymatic antioxidants total phenols and flavonoid, water-soluble soluble antioxidant metabolites expressed as ascorbic acid; the antioxidant potential of hyssop methanol extracts assessed by two common methods: free radical scavenging activity using free stable radical (2,2-diphenyl-1-picrylhydrazyl, DPPH• and ferric reducing antioxidant power FRAP in flowers and leaves. The successfully adapted to field conditions in vitro plants (survival rate 95%) were inoculated with arbuscular mycorrhizal fungi (Claroideoglomus claroideum, ref. EEZ 54). It was established that the activities of enzymes with antioxidant capacity (superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase) were significantly higher in leaves than in flowers in both control and mycorrhized plants. In flowers and leaves of inoculated plants, the antioxidant enzymes activity were lower than in non-inoculated plants, only in SOD activity, there was no difference. The content of low molecular metabolites with antioxidant capacity as total phenols, total flavonoids, and water soluble antioxidants was higher in inoculated plants. There were no significant differences between control and inoculated plants both for FRAP and DPPH antioxidant activity. According to plant essential oil content, there was no difference between non-inoculated and inoculated plants. Based on our results we could suggest that antioxidant capacity of in vitro propagated hyssop plant under conditions of cultivation is determined by the phenolic compounds-total phenols and flavonoids as well as by the levels of water-soluble metabolites with antioxidant potential. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.

Keywords: antioxidant enzymes, antioxidant metabolites, arbuscular mycorrhizal fungi, Hyssopus officinalis L.

Procedia PDF Downloads 328

Authors: Serap Gedikli, Hakan Çakmak, M. Buğra Güldiken, Duygu Yalnızoğlu

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Air, which is necessary for living things to survive; while it carries some useful substances in it, it can also carry foreign particles of different sizes that may be harmful to the health. All airborne organic substances of biological origin, including bacteria, fungi, fungal spores, viruses, pollen, and their components, are called "bioaerosols". Nowadays, everyone spends most of their time in closed areas such as home, workplace, school, etc. Although it is known that outdoor air pollution affects health, it is not known that indoor air pollution has harmful effects in terms of health. In this study, indoor air microbial load and SARS-CoV-2 virus cleaning efficiency of Froumann brand air cleaners were studied. This work in 300 m³, 600 m³, and 1000 m³ completely closed areas without any air circulation with Froumann N80, N90, and N100 air-cleaning devices. Analyzes were performed for both areas at 60 minutes before and after the device was operated using a particle measuring device (Particles Plus 7302) and an air sampler (Mas-100 ECO). The measurements were taken by placing the test equipment 1.5-2 m away from the air cleaner. At the same time, the efficiency of the HEPA filter was evaluated by taking samples from the air outlet point of the HEPA filter using the air sampling device (Mas-100 ECO) after the device was started. Nutrient agar and malt agar are used as total mesophilic bacteria and total fungi. The number of colony-forming units per m³ (cfu/m³) was calculated by counting colonies in Petri dishes after incubation for 48 hours at 37°C for bacteria and 72 hours at 30°C for fungi. The change in the number of colonies and the decrease in the microbial load was calculated as a percentage value. SARS-CoV-2 activity analysis studies were carried out by İnönü University Microbiology Department in accordance with the World Health Organization regulations. Finally, the HEPA filter in the devices used was taken and kept under a certain temperature and humidity, and the change in the microbial load on it was monitored over a 6-month period. At the end of the studies, a 91%-94% reduction was determined in the total mesophilic bacteria count of Frouman brand N80, N90, and N100 model air cleaners. A decrease of 94%-96% was detected in the total number of yeast/molds. HEPA filter efficiency was evaluated, and at the end of the analysis, 98% of the bacterial load and approximately 100% of yeast/mold load at the HEPA filter air outlet point were decreased. According to the SARS- CoV-2 analysis results, when the device is operating at the medium airflow level 3, it can filter virus-carrying aerosols by 99%. As a result, it was determined that the Froumann model air cleaner was effective in controlling and reducing the microbial load in the indoor air.

Keywords: HEPA filter, indoor air quality, microbial load, SARS-CoV-2

Procedia PDF Downloads 205

Authors: Sangyeop Shin, D. C. M. Kulatunga, S. H. S. Dananjaya, Chamilani Nikapitiya, Jehee Lee, Mahanama De Zoysa

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Saprolegniasis is one of the most devastating fungal diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated fungi belong to the member of S. parasitica, supported by its typical fungal features including cotton-like whitish mycelium, zoospores (primary and secondary) and phylogenetic analysis with internal transcribed spacer (ITS) region. Pathogenicity of isolated S. parasitica was developed in embryo, larvae, juvenile and adult zebrafish as a disease model. Up regulation of host genes encoding ZfTnf-α, Zfc-Rel, ZfIl-12, ZfLyz-c, Zfβ-def, and ZfHsp-70 was identified in zebrafish larvae after experimental challenge of S. parasitica showing the host immune responses against the S. parasitica. Survival of the juveniles upon fungal infection might be due to the increased immune protection in the host. Investigation of antifungal susceptibility of S. parasitica with natural lawsone (2-hydroxy-1,4-naphthoquinone) revealed the minimum inhibitory concentration (MIC) and percentage inhibition of radial growth (PIRG %) as 200 µg/mL and 31.8%, respectively. Lawsone was able to change the membrane permeability, and cause irreversible damage and disintegration to the cellular membranes of S. parasitica which might have effect on fungi growth inhibition. Moreover, the mycelium exposed to lawsone (MIC level) changed the transcriptional responses of S. parasitica genes. Overall results indicate that lawsone could be a potential and novel anti-S. parasitica agent for controlling S. parasitica infection.

Keywords: host-pathogen interactions, lawsone, rainbow trout, Saprolegnia parasitica, Saprolegniasis, zebrafish

Procedia PDF Downloads 252

Authors: Ahmed Samy Elnoby

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Ailanthus altissima native to Asia which belongs to the family Simaroubaceae was subjected to phytochemical screening and biological investigations. Phytochemical screening revealed the presence of carbohydrates, tannins, sterols, flavonoids and traces of saponins. In addition, quantitative determination of phenolics and flavonoid content were performed. The antimicrobial activity of methanolic extract of the leaves was determined against gram-positive, gram-negative bacteria in addition to fungi using a modified Kirby-Bauer disc diffusion method that was compared with standard discs ampicillin which acts as an antibacterial agent and amphotericin B which acts as an antifungal agent. A high potency was observed against gram-positive bacteria mainly staphylococcus aureus, gram-negative bacteria mainly Escherichia coli and showed no potency against fungi mainly Aspergillus flavus and candida albicans. On the other hand, the antioxidant activity of the extract was determined by 1, 1-diphenyl-2- diphenyl-2-picryl-hydrazil (DPPH). A very low potency was shown by using DPPH for the antioxidant effect so IC50 = 0 ug/ml, IC90 =0 ug /ml and remark gave 47.2 % at 100 ug/ml which is very weak. Cytotoxic activity was determined by using MTT assay (3-4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide) against MCF7 (Human Caucasian breast adenocarcinoma) cell line. A moderate potency was shown by using MCF7 cell line for cytotoxic effect so LC50= 90.2 ug/ml, LC90=139.9 ug/ml and the remark gave 55.2% at 100 ug/ml which is of moderate activity so, Ailanthus altissima can be considered to be a promising antimicrobial agent from natural origin.

Keywords: Ailanthus altissima, TLC, HPLC, anti-microbial activity, antifungal activity, antioxidant, cytotoxic activity

Procedia PDF Downloads 175

Authors: Howaida I. Abd-Alla, Amal Z. Hassan, Maha Soltan, Atef G. Hanna, Mounir M. El-Safty

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Acokanthera oblongifolia (Apocynaceae) is used for treatment of several infection diseases and is a well-known cardiac glycoside-containing plant. The infusion of their leaves is gargled to treat tonsillitis and is used medicinally to treat snakebites. The total cardiac glycosides content in the leaves was determined by referring to gitoxigenin as a reference compound. Two triterpenes, lup-20(29)-en-3β-ol (1) and oleanolic acid (2); two cardenolides, acovenoside A (3) and acobioside A (4) were isolated from the ethyl acetate extract. Their structures were determined on the basis of spectral analysis. Major constituents isolated from this species were evaluated for cytotoxicity against normal lung cell line (Wi38) and antimicrobial activities against Gram-positive (two strains) and Gram-negative bacteria (four strains), yeast-like fungi (two strains) and fungi (five strains). The minimum inhibitory concentration (MIC) of the compounds was determined using broth microdilution method. Their viral inhibitory effects against avian influenza virus type A (AI-H5N1) and Newcastle disease virus (NDV) in specific pathogen free (SPF) embryonated chicken eggs (ECE), chicken embryo fibroblasts (CEF) and Vero cells were evaluated. The cardenolide (3) showed viral inhibitory effects against AI-H5N1 and NDV in SPF ECE. The two cardenolides isolated have shown potent cytotoxicity against Vero cells. Compound (3) showed potent anti-Gram-negative bacteria activity. These results suggested that acovenoside A might be promising for future antiviral and antimicrobial drug design.

Keywords: Acokanthera, AI-H5N1, Cardenolides, NDV, SPF-ECE, VERO, Wi38 , Microbe

Procedia PDF Downloads 179

Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

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In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: trichoderma spp., meloidogyne javanica, Fusarium oxysporum f.sp.radicis lycopersici, biocontrol

Procedia PDF Downloads 279

Authors: Deepanwita Deka, Dhruva Kumar Jha

Abstract:

Endophytes are the microbes that colonize living, internal tissues of plants without causing any immediate, overt negative effects. Endophytes are rich source of therapeutic substances like antimicrobial, anticancerous, herbicidal, insecticidal, immunomodulatory compounds. Brucea mollis, commonly known as Quinine in Assam, belonging to the family Simaroubaceae, is a shrub or small tree, recorded as endangered species in North East India by CAMP survey in 2003. It is traditionally being used as antimalarial and antimicrobial agent and has antiplasmodial, cytotoxic, anticancer, diuretic, cardiovascular effect etc. Being endangered and medicinal; this plant may host certain noble endophytes which need to be studied in depth. The aim of the present study was isolation and identification of potent endophytic fungi from Brucea mollis, an endangered medicinal plant, to protect it from extinction due to over use for medicinal purposes. Aseptically collected leaves, barks and roots samples of healthy plants were washed and cut into a total of 648 segments of about 2 cm long and 0.5 cm broad with sterile knife, comprising 216 segments each from leaves, barks and roots. These segments were surface sterilized using ethanol, mercuric chloride (HgCl2) and aqueous solution of sodium hypochlorite (NaClO). Different media viz., Czapeck-Dox-Agar (CDA, Himedia), Potato-Dextrose-Agar (PDA, Himedia), Malt Extract Agar (MEA, Himedia), Sabourad Dextrose Agar (SDA, Himedia), V8 juice agar, nutrient agar and water agar media and media amended with plant extracts were used separately for the isolation of the endophytic fungi. A total of 11 fungal species were recovered from leaf, bark and root tissues of B. mollis. The isolates were screened for antimicrobial, antioxidant and enzymatic activities using certain protocols. Cochliobolus geniculatus was identified as the most dominant species. The mycelia sterilia (creamy white) showing highest inhibitory activity against Candida albicans (MTCC 183) was induced to sporulate using modified PDA media. The isolate was identified as Geosmithia pallida. The internal transcribed spacer of rDNA was sequenced for confirmation of the taxonomic identity of the sterile mycelia (creamy white). The internal transcribed spacer r-DNA sequence was submitted to the NCBI (KU693285) for the first time from India. G. pallida and Penicillium showed highest antioxidant activity among all the isolates. The antioxidant activity of G. pallida and Penicillium didn’t show statistically significant difference (P˃0.05). G. pallida, Cochliobolus geniculatus and P. purpurogenum respectively showed highest cellulase, amylase and protease activities. Thus, endopytic fungal isolates may be used as potential natural resource of pharmaceutical importance. The endophytic fungi, Geosmithia pallida, may be used for synthesis of pharmaceutically important natural products and consequently can replace plants hitherto used for the same purpose. This study suggests that endophytes should be investigated more aggressively to better understand the endophyte biology of B. mollis.

Keywords: Antimicrobial activity, antioxidant activity, Brucea mollis, endophytic fungi, enzyme activity, Geosmithia pallida

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