Search results for: fungal endophyte

Authors: Gómez S. Jennifer, Méndez V. Camila, Moncayo M. Diana, Vega M. Lizeth

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The cape gooseberry is a climacteric fruit; Colombia is one of the principal exporters in the world. The environmental condition of temperature and relative moisture decreases the titratable acidity and pH. These conditions and fruit maturation result in the fungal proliferation of Botrytis cinerea disease. Plastic packaging for fresh cape gooseberries was used for mechanical damage protection but created a suitable atmosphere for fungal growth. Beta-cyclodextrins are currently implemented as coatings for the encapsulation of hydrophobic compounds, for example, with bioactive compounds from essential oils such as cinnamaldehyde, which has a high antimicrobial capacity. However, it is a volatile substance. In this article, the casting method was used to obtain a polylactic acid (PLA) polymer film containing the beta-cyclodextrin-cinnamaldehyde inclusion complex, generating an insert that allowed the controlled release of the antifungal substance in packed cape gooseberries to decrease contamination by Botrytis cinerea in a latent state during storage. For the encapsulation technique, three ratios for the cinnamaldehyde: beta-cyclodextrin inclusion complex were proposed: (25:75), (40:60), and (50:50). Spectrophotometry, colorimetry in L*a*b* coordinate space and scanning electron microscopy (SEM) were made for the complex characterization. Subsequently, two ratios of tween and water (40:60) and (50:50) were used to obtain the polylactic acid (PLA) film. To determine mechanical and physical parameters of colourimetry in L*a*b* coordinate space, atomic force microscopy and stereoscopy were done to determine the transparency and flexibility of the film; for both cases, Statgraphics software was used to determine the best ratio in each of the proposed phases, where for encapsulation it was (50:50) with an encapsulation efficiency of 65,92%, and for casting the ratio (40:60) obtained greater transparency and flexibility that permitted its incorporation into the polymeric packaging. A liberation assay was also developed under ambient temperature conditions to evaluate the concentration of cinnamaldehyde inside the packaging through gas chromatography for three weeks. It was found that the insert had a controlled release. Nevertheless, a higher cinnamaldehyde concentration is needed to obtain the minimum inhibitory concentration for the fungus Botrytis cinerea (0.2g/L). The homogeneity of the cinnamaldehyde gas phase inside the packaging can be improved by considering other insert configurations. This development aims to impact emerging food preservation technologies with the controlled release of antifungals to reduce the affectation of the physico-chemical and sensory properties of the fruit as a result of contamination by microorganisms in the postharvest stage.

Keywords: antifungal, casting, encapsulation, postharvest

Procedia PDF Downloads 46

Authors: Gunjan Srivastava, Shamila Kalia

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Dalbergia sissoo Roxb., (Family- Leguminosae; Subfamily- Papilionoideae), is an economically and ecologically important tree species having medicinal value. Of the rich complex of insect fauna, ten have been recognized as potential pests of nurseries and plantations. Present study was conducted to explore an effective ecofriendly control of Dichomeris eridantis Meyrick, an important defoliator pest of D. sissoo. Health and environmental concerns demanded devising a bio-intensive pest management strategy and employing ecofriendly measures. In the present laboratory bioassay two entomopathogenic fungi Acremonium perscinum and Beauveria bassiana were tested and compared for evaluating the efficacy of their seven different concentrations (besides control) against the 3rd, 4th and 5th instar larvae of D. eridantis, on the basis of mean percent mortality data recorded and tabulated for seven days after treatment application. Analysis showed that both treatments vary significantly among themselves. Also, variations amongst instars and duration with respect to their mortality were highly significant (p < .001). All their interactions were found to vary significantly. B. bassiana at 0.25x107 spores / ml spore concentration caused maximum mean percent mortality (62.38%) followed by mean percent mortality at its 0.25x106 spores / ml concentration (56.67%). Mean percent mortality at maximum spore concentration (0.054x107 spores / ml) and next highest spore concentration (0.054 x106 spores / ml) due to A. perscinum treatment were far less effective (mean percent mortality of 45.40% and 31.29%, respectively). At 168 hours mean percent mortality of larval instars due to both fungal treatment applications reached its maximum (52.99%) whereas, at 24 hours mean percent mortality remained least (5.70%). In both cases, treatments were most effective against 3rd instar larvae and least effective against 5th instar larvae. A comparative acccount of efficacy of B. bassiana and A. perscinum on the 3rd, 4th and 5th instar larvae of D. eridantis on 5th, 6th and 7th post treatment observation days after their application, on the basis of their median lethal concentrations (LC50) proved B. bassiana to be more potential microbial pathogen of the two fungal microbes, for all the three instars (3rd, 4th and 5th) of D. eridantis, on all the three days (5th, 6th and 7th post observation days after application of both treatments). Percent mortality of D. eridantis increased in a dose dependent manner. Koch’s Postulates tested positive, thus confirming the pathogenicity of B. bassiana against the larval instars of D. eridantis. LC90 values of 0.280x1011 spores/ml, 0.301x108 spores/ml and 0.262x108 spores/ml concentrations of B. bassiana were standardized which can effectively cause mortality of all the larval instars of D. eridantis in the field after 5th, 6th and 7th day of their application, respectively. Therefore, these concentrations can be safely used in nurseries as well as plantations of D. sissoo for effective control of D. eridantis larvae.

Keywords: Acremonium perscinum, Beauveria bassiana, Dalbergia sissoo, Dichomeris eridantis

Procedia PDF Downloads 201

Authors: Ramdane Gaouir

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Farmers in the two regions of Tizi-Ouzou, DBK and Tadmait, face different crop pests, namely, fungal diseases, insect attack as well as different types of deficiencies. To fight against all these threats, they resort to the use of a wide variety of chemicals. Our survey carried out in these two regions, aims to identify the different types of pesticides used, the method of management of the packaging generated by these phytosanitary products as well as the evaluation of the effect of these products on the farmers' health and the environment. The results obtained highlighted the use of two types of pesticides with a very wide application, namely fungicides and insecticides. The two most widely identified chemical families are neonicotoids and organophosphates, which are among the most toxic and persistent pesticides in the environment. The management method of packaging and the application of products by spraying reflect the lack of training and the unawareness of farmers vis-à-vis the impact of pesticides on their health and on the environment.

Keywords: farmers, crops, pesticides, fungicides, neonicotinoids, organochlorines

Procedia PDF Downloads 53

Authors: N. Bakradze, T. Dumbadze, N. Gagelidze, L. Amiranashvili, A. D. L. Batako

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Cereals are considered as a strategic product in human life and it demand is increasing with the growth of world population. There is always shortage of cereals in various areas of the globe. For example, Georgia own production meets only 15-20% of the demand for grain, despite the fact that the country is considered one of the main centers of wheat origin. In Georgia, there are 14 types of wheat and more than 150 subspecies, and 40 subspecies of common wheat. Increasing wheat production is important for the country. One of the ways to solve the problem is to develop and implement new, environmentally and economically acceptable technologies. Such technologies include pre-sowing treatment of seed with a laser and associative nitrogen-fixing of the Azospirillum brasilensse bacteria. In the region there are Dika and Lomtagora which are among the most common in Georgia. Dika is a frost-resistant wheat, with a high ability to adapt to the environment, resistant to falling and it is sown in highlands. Dicka excellent properties are due to its strong immunity to fungal diseases; Dicka grains are rich in protein and lysine. Lomtagora 126 differs with its winter and drought resistance, and, it has a great ability to germinate. Lomtagora is characterized by a strong root system and a high budding capacity. It is an early variety, fall-resistant, easy to thresh and suitable for mechanized harvesting with large and red grains. The plant is moderately resistant to fungal diseases. This paper presents some preliminary experimental results where, a continuous CO2 laser at a power of 25-40 W/cm2 was used to radiate grains at a flow rate of 10-15 cm/sec. The treatment was carried out on grains of the Triticum aestivum L. var. of Lutescens (local variety name - Lomtagora 126), and Triticum carthlicum Nevski (local variety name - Dika). Here the grains were treated with Azospirillum brasilensse isolate (108-109 CFU / ml), which was isolated from the rhizosphere of wheat. It was observed that the germination of the wheat was not significantly influenced by either laser or bacteria treatment. In the case of the variety Lomtagora 126, when irradiated at an angle of 90°, it slightly improved the growth within 38 days of sawing, and in the case of irradiation at an angle of 90°+1, by 23%. The treatment of seeds with Azospirillum brazilense in both irradiated and non-irradiated variants led to an improvement in the growth of ssprouts. However, in the case of treatment with azospiril alone - by 22%, and with joint treatment of seeds with azospiril and irradiation - by 29%. In the case of the Dika wheat, the irradiation only led to an increase in growth by 8-9%, and the combine treatment of seeds with azospiril and irradiation - by 10-15%, in comparison with the control. Thus, the combine treatment of wheat of different varieties provided the best effect on the growth. Acknowledgment: This work was supported by Shota Rustaveli National Science Foundation of Georgia (SRNSFG) (Grant number CARYS 19-573)

Keywords: laser treatment, Azospirillum brasilensse, seeds, wheat varieties, Lomtagora, Dika

Procedia PDF Downloads 114

Authors: Abdulmajeed Bashir Mlitan, Ethan Hack

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Soil samples were collected from and around Alkomos cement factory, Alkomos town, Libya. Soil physiochemical properties were determined. In addition, olive leaves were scanned for their fungal content. This work can conclude that the results obtained for the examined physiochemical characteristics of soil in the area studied prove that cement dust from the Alkomos cement factory in Libya has had a significant impact on the soil. The affected soil properties are pH and total calcium content. These characteristics were found to be higher than those in similar soils from the same area. The increment of soil pH in the same area may be a result of precipitation of cement dust over the years. Different responses were found in each season and each site. For instance, the dominance of fungi of soil and leaves was lowest at 100 m from the factory and the evenness and diversity increased at this site compared to the control area and 250 m from the factory.

Keywords: pollution, soil microbial, alkomos, Libya

Procedia PDF Downloads 581

Authors: H. Heydari, B. Konuklugil, P. Proksch

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Marine-derived fungi can produce interesting bioactive secondary metabolites that can be considered the potential for drug development. Turkey is a country of a peninsula surrounded by the Black Sea at the north, the Aegean Sea at the west, and the Mediterranean Sea at the south. Despite the approximately 8400 km of coastline, studies on marine secondary metabolites and their biological activity are limited. In our ongoing search for new natural products with different bioactivities produced by the marine-derived fungi, we have investigated secondary metabolites of Turkish collection of the marine sea slug (Peltodoris atromaculata) associated fungi Hypocrea nigricans collected from Seferihisar in the Egean sea. According to the author’s best knowledge, no study was found on this fungal species in terms of secondary metabolites. Isolated from ethyl acetate extract of the culture of Hypocrea nigricans were (isodihydroauroglaucin,tetrahydroauroglaucin and dihydroauroglaucin. The structures of the compounds were established based on an NMR and MS analysis. Structural elucidation of another isolated secondary metabolite/s continues.

Keywords: Hypocrea nigricans, isolation, marine fungi, secondary metabolites

Procedia PDF Downloads 132

Authors: Abdulaziz Bashir Kutawa1, 2, *, Khairulmazmi Ahmad 1, 3, Mohd Zobir Hussein 4, Asgar Ali 5, * Mohd Aswad Abdul Wahab1, Amara Rafi3, Mahesh Tiran Gunasena1, 6, Muhammad Ziaur Rahman1, 7, Md Imam Hossain1, And Syazwan Afif Mohd Zobir1

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Various abiotic and biotic stresses have an impact on rice production all around the world. The most serious and prevalent disease in rice plants, known as rice blast, is one of the major obstacles to the production of rice. It is one of the diseases that has the greatest negative effects on rice farming globally, the disease is caused by a fungus called Magnaporthe grisae. Since nanoparticles were shown to have an inhibitory impact on certain types of fungus, nanotechnology is a novel notion to enhance agriculture by battling plant diseases. Utilizing nanocarrier systems enables the active chemicals to be absorbed, attached, and encapsulated to produce efficient nanodelivery formulations. The objectives of this research work were to determine the efficacy and mode of action of the nanofungicides (in-vitro) and in field conditions (in-vivo). Ionic gelation method was used in the development of the nanofungicides. Using the poisoned media method, the synthesized agronanofungicides' in-vitro antifungal activity was assessed against M. grisae. The potato dextrose agar (PDA) was amended in several concentrations; 0.001, 0.005, 0.01, 0.025, 0.05, 0.1, 0.15, 0.20, 0.25, 0.30, and 0.35 ppm for the nanofungicides. Medium with the only solvent served as a control. Every day, mycelial growth was measured, and PIRG (percentage inhibition of radial growth) was also computed. Every day, mycelial growth was measured, and PIRG (percentage inhibition of radial growth) was also computed. Based on the results of the zone of inhibition, the chitosan-hexaconazole agronanofungicide (2g/mL) was the most effective fungicide to inhibit the growth of the fungus with 100% inhibition at 0.2, 0.25, 0.30, and 0.35 ppm, respectively. Then followed by carbendazim analytical fungicide that inhibited the growth of the fungus (100%) at 5, 10, 25, 50, and 100 ppm, respectively. The least were found to be propiconazole and basamid fungicides with 100% inhibition only at 100 ppm. The scanning electron microscope (SEM), confocal laser scanning microscope (CLSM), and transmission electron microscope (TEM) were used to study the mechanisms of action of the M. grisae fungal cells. The results showed that both carbendazim, chitosan-hexaconazole, and HXE were found to be the most effective fungicides in disrupting the mycelia of the fungus, and internal structures of the fungal cells. The results of the field assessment showed that the CHDEN treatment (5g/L, double dosage) was found to be the most effective fungicide to reduce the intensity of the rice blast disease with DSI of 17.56%, lesion length (0.43 cm), DR of 82.44%, AUDPC of 260.54 Unit2, and PI of 65.33%, respectively. The least treatment was found to be chitosan-hexaconazole-dazomet (2.5g/L, MIC). The usage of CHDEN and CHEN nanofungicides will significantly assist in lessening the severity of rice blast in the fields, increasing output and profit for rice farmers.

Keywords: chitosan, hexaconazole, disease incidence, and magnaporthe grisae

Procedia PDF Downloads 39

Authors: W. Lakhdari, B. Doumendji-Mitich, A. Dahliz, S. Doumendji, Y. Bouchikh, R. M'lik, H. Hammi, A. Soud

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This work was done in order to fight against Brachytrupes megacephalus, a major pest in the Algerian oasis and promote one aspect of biological control against it. He wears a hand on the isolation and identification of indigenous fungi on imagos of this insect harvested in the station of INRAA Touggourt and secondly, the study of the pathogenicity of these strains fungal on this orthoptère adults. The results obtained showed the presence of six different species of entomopathogenic fungi, it is: Aspergillus flavus, Fusarium sp, Beauveria bassiana, Penicillium sp, Metharizium anisopliae and Aspergillus Niger. The pathogenicity test using fungi Beauveria bassiana strains and Metharizium anisopliae. On adult of B. megacephalus highlights the effectiveness of these strains of predatory adults, with a mortality rate approaching 100% after 11 days.

Keywords: biological control, brachytrupes megacephalus, entomopathogenic fungi, Southeastern Algeria

Procedia PDF Downloads 386

Authors: Blessing Atim Aderibigbe

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A series of amine/ester-linked hybrid compounds containing pharmacophores, such as ursolic acid, oleanolic acid, ferrocene and bisphosphonates, were synthesized in an attempt to develop potent antibacterial and anticancer agents. Their structures were analyzed and confirmed using Nuclear Magnetic Resonance, Fourier Transform Infrared Spectroscopy, and mass spectroscopy. All the synthesized hybrid compounds were evaluated for their antibacterial activities against eleven selected bacterial strains using a serial dilution method. Some of the compounds displayed significant antibacterial activity against most of the bacterial and fungal strains. In addition, the in vitro cytotoxicity of these compounds was also performed against selected cancer cell lines. Some of the compounds were also found to be more active than their parent compounds, revealing the efficacy of designing hybrid molecules using plant-based bioactive agents.

Keywords: ursolic acid, hybrid drugs, oleanolic acid, bisphosphonates

Procedia PDF Downloads 52

Authors: Payam Rasoulnia, Seyyed Mohammad Mousavi

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Heavy fuel oil firing power plants produce huge amounts of ashes as solid wastes, which seriously need to be managed and processed. Recycling precious metals of V and Ni from these oil-fired ashes which are considered as secondary sources of metals recovery, not only has a great economic importance for use in industry, but also it is noteworthy from the environmental point of view. Vanadium is an important metal that is mainly used in the steel industry because of its physical properties of hardness, tensile strength, and fatigue resistance. It is also utilized in oxidation catalysts, titanium–aluminum alloys and vanadium redox batteries. In the present study bioleaching of vanadium and nickel from an oil-fired ash sample was conducted using Aspergillus niger fungus. The experiments were carried out using spent-medium bioleaching method in both Erlenmeyer flasks and also bubble column bioreactor, in order to compare them together. In spent-medium bioleaching the solid waste is not in direct contact with the fungus and consequently the fungal growth is not retarded and maximum organic acids are produced. In this method the metals are leached through biogenic produced organic acids present in the medium. In shake flask experiments the fungus was cultured for 15 days, where the maximum production of organic acids was observed, while in bubble column bioreactor experiments a 7 days fermentation period was applied. The amount of produced organic acids were measured using high performance liquid chromatography (HPLC) and the results showed that depending on the fermentation period and the scale of experiments, the fungus has different major lixiviants. In flask tests, citric acid was the main produced organic acid by the fungus and the other organic acids including gluconic, oxalic, and malic were excreted in much lower concentrations, while in the bioreactor oxalic acid was the main lixiviant and it was produced considerably. In Erlenmeyer flasks during 15 days fermentation of Aspergillus niger, 8080 ppm citric acid and 1170 ppm oxalic acid was produced, while in bubble column bioreactor over 7 days of fungal growth, 17185 ppm oxalic acid and 1040 ppm citric acid was secreted. The leaching tests using the spent-media obtained from both of fermentation experiments, were performed at the same conditions of leaching duration of 7 days, leaching temperature of 60 °C and pulp density up to 3% (w/v). The results revealed that in Erlenmeyer flask experiments 97% of V and 50% of Ni were extracted while using spent medium produced in bubble column bioreactor, V and Ni recoveries were achieved to 100% and 33%, respectively. These recovery yields indicate that in both scales almost total vanadium can be recovered, while nickel recovery was lower. With help of the bioreactor spent-medium nickel recovery yield was lower than that of obtained from the flask experiments, which it could be due to precipitation of some values of Ni in presence of high levels of oxalic acid existing in its spent medium.

Keywords: Aspergillus niger, bubble column bioreactor, oil-fired ash, spent-medium bioleaching

Procedia PDF Downloads 209

Authors: Adferotimi Banso

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A laboratory investigation was conducted to determine the effect of Justica flava and Aframonium melegueta on the growth of Aspergillus niger, Rhizopus stolonifer and Fusarium species in sweet orange and mango juices. Aqueous extract (3%v/v) of Justica flava and Aframonium melegueta reduced the growth of the fungi, a combination of 2% (v/v) each of Justica flava and Aframonium melegueta extracts reduced the growth better. Partial purification of aqueous extracts of Justica flava and Aframonium melegueta showed that ethyl acetate fraction of the extracts exhibited the highest level of inhibition of growth of the test fungi compared with diethyl ether and n-hexane fractions. The results suggest that extracts of Justica flava and Aframonium melegueta may be important substitutes for conventional chemical preservatives in the processing of fruit juices.

Keywords: aqueous, fraction, mango, orange, purification, sweet

Procedia PDF Downloads 317

Authors: Reenu Yadav, S. K. Yadav

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Herbal therapy has been used for daily oral health care to prevent, treat or cure oral conditions from halitosis to periodontal diseases. The importance of mouth and teeth cleanliness has been recognized from the earliest days of civilization to the 21st century. In the present study, leaves and seeds of Cordia obliqua and barks and twigs of Beautea monosperma, which is used traditionally for oral diseases was evaluated for its antimicrobial activity. The antimicrobial activity tests indicated that the methanolic extract exhibited stronger activities against the commonly encountered oral bacterial and fungal pathogens. The mouthwash formulation prepared and it is compared with marketed formulation HiOra. The results indicated that the herbal mouthwash could inhibit the growth of oral pathogens and may prevent plaque and other periodontal diseases caused by dental pathogens.

Keywords: herbal mouthwash, bio medicine, life sciences, herbal extracts

Procedia PDF Downloads 313

Authors: Abdelaziz Wided, Khiat Nawel, Khiat Inssaf

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The present study was initiated to: Determine burned forest-inhabiting fungi in Zouagha, Terri Beinène, Mila and study the antagonistic activity of Trichoderma sp against Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp. 18 fungal strains were isolated from Soil samples taken from the forest Zouagha (Burned) in the region Mila representing 6 genera: Trichoderma sp et Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp, Rhizopus sp. The tests of dual culture method on culture medium (PDA) against Trichoderma sp et Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp revealed that: Trichoderma sp could reduce l mycelium grouth of Fusarium sp23.13%, Penicillium sp33.13%, Rhizoctoniasp33.75 %and Alternaria sp 38.31% in comparaison with the witness after 6 days at room temperature. The strains of Fusarium sp ,Penicillium sp, Rhizoctonia sp et Alternaria sp showed differences sensibility to the antagoniste.

Keywords: isolation, identification, molds, burned soil of zouagha, antagonism, trichoderma sp

Procedia PDF Downloads 224

Authors: Sahira N. Muslim, Alaa N. Mohammed, Saba Saadoon Khazaal, Batool Kadham Salman, Israa M. S. AL-Kadmy, Sraa N. Muslim, Ahmed S. Dwaish, Sawsan Mohammed Kareem, Sarah N. Aziz, Ruaa A. Jasim

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Tannase has wide applications in food, beverage, brewing, cosmetics and chemical industries and one of the major applications of tannase is the production of gallic acid. Gallic acid is used for manufacturing of trimethoprim. In the present study, a local fungal strain of Penicillium expansum A₄ isolated from spoilt apple samples gave the highest production level of tannase. Tannase was partially purified with a recovery yield of 92.52% and 6.32 fold of purification by precipitation using ammonium sulfate at 50% saturation. Tannase led to increased antimicrobial activity of ceftazidime against Pseudomonas aeruginosa and S. aureus and had a synergism effect at low concentrations of ceftazidime, and thus, tannase may be a useful adjuvant agent for the treatment of many bacterial infections in combination with ceftazidime.

Keywords: ceftazidime, Penicillium expansum, tannase, antimicrobial activity

Procedia PDF Downloads 700

Authors: Jamal Elzwai, Kofi Aidoo, Alan Candlish

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Production of OTA was detected after 24hr by Aspergillus ochraceus isolate whereas at 36hr for A. carbonarius isolate and Penicillium verrucosum IMI 285522 and 60hr for A. ochraceus CBS 588.68. Highest OTA level was produced by A. carbonarius isolate followed by A. ochraceus CBS 588.68, Penicillium verrucosum IMI 285522 and finally A. ochraceus isolate. Glucosamine content of barley sample before fermentation was found to be negligible and remained almost constant during the incubation time. Glucosamine content started to increase at 12 hours after incubation with A. ochraceus isolate, A. carbonarius isolate and A. ochraceus CBS 588.68, and after 12 hours with P. verrucosum IMI 285522. Highest glucosamine content, as a result of increase in fungal biomass, was produced by A. ochraceus CBS 588.68 followed by A. ochraceus isolate, A. carbonarius isolate, and finally by P. verrucosum IMI 285522. It appears that there is a correlation between OTA synthesis and glucosamine content with A. ochraceus isolate, A. carbonarius isolate and A. ochraceus CBS 588.68 but not with P. verrucosum IMI 285522.

Keywords: chitin, barley, Ochratoxin A, Aspergiluus ochraceus, A. carbonarius, Penicillium verrucosum

Procedia PDF Downloads 400

Authors: Hye-Hyun Lee, Hyo-Kon Chun, Kyung-Hee Kim Kim, Mi-Hee Kim, Saet-Byul Chu, Sang-Jong Lee, Seung-Hyeop Lee, Seung-Won Yi

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Some chemicals such as malachite green, methylene blue, and copper sulfate had been used frequently as disinfectants controlling fungal infection in aquaculture. However, their carcinogenicity, mutagenicity and teratogenicity were reported in mammals. After their accumulation in food fish and its consumers was confirmed, concerns about public health has resulted in enhanced monitoring and increased demand for eco-friendly treatments. Therefore, this study aimed to evaluate safety to fish and efficacy of sulfur solution processed by effective microorganisms (EM-PSS) against Saprolegnia parasitica, for use of a potential aquatic fungicidal disinfectant. The natural sulfur purchased from Kawah Ijen volcano, East Java, Indonesia was processed by the liquid mixture consisting of following twelve effective microorganisms (Rapha-el®; Lbiotech, Jeonnam, Korea), Lactobacillus parafarraginis, L. paracasei, L. harbinensis, L. buchneri, L. perolens, L. rhamnosus, L. vaccinostercus, Acetobacter lovaniensis, A. peroxydans, Pichia fermentans, Candida ethanolica, Saccharomycopsis schoenii isolated from fermentation process of oriental medicinal herbs including green tea, privet, and puer tea. The material was applied to in vitro antifungal activity test for Saprolegnia parasitica using agar dilution method. In addition, an acute toxicity test was performed on carp (Cyprinus carpio), eel (Anguilla japonica), and mud fish (Misgurnus mizolepis) for 96 hours. After three species of fish (n=15) were accustomed to experimental water environment for three days, the EM-PSS was added to each tank as final concentrations to be 0 to 500 ppm. The fish were taken into necropsy, and the histological sections of the gill, liver, and spleen were counter-stained with hematoxylin and eosin (H-E). And hence, no observed effect concentration (NOEC) of the solution was used for taking a medicinal bath for mudfish infected by Saprolegnia parasitica in practice. The result of in vitro antifungal activity test showed the growth inhibition of the fungus at 100 ppm, which and the lower concentrations occurred no fatal case in any fish species tested until the end of the examination. The 125 ppm of the solution, however, resulted in 13.3 %, 13.3 %, and 6.3 % of mortality in carp, eel, and mudfish, respectively. But both 250 and 500 ppm of the solution leaded lethality to all population of each fish species within 24 hours. Besides, H-E staining also showed no specific evidence for toxicity in fish at lesser than 100 ppm of EM-PSS. On the other hand, as a result of field application of the solution, no growth of fungal mycelium was found in fish bodies from gross observation 5 days post treatment. In conclusion, 100ppm of EM-PSS resulted in inhibition and treatment of Saprolegnia parasitica infection. In addition, the use of EM-PSS lower than 100 ppm is safe for fish. Therefore, EM-PSS could be used as aquatic fungicide, and also may be possible to be a potential eco-friendly disinfectant in aquaculture.

Keywords: antifungal activity, effective microorganism, toxicity, saprolegnia, processed sulfur solution

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Authors: Aitor Barbero-López, Soumaya Chibily, Gerhard Scheepers, Thomas Grahn, Martti Venäläinen, Antti Haapala

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Wood decay is addressed continuously within the wood industry through use and development of wood preservatives. The increasing awareness on the negative effects of many chemicals towards the environment is causing political restrictions in their use and creating more urgent need for research on green alternatives. This paper discusses some of the possible natural extracts for wood preserving applications and compares the analytical methods available for testing their behavior and efficiency against decay fungi. The results indicate that natural extracts have interesting chemical constituents that delay fungal growth but vary in efficiency depending on the chemical concentration and substrate used. Results also suggest that presence and redistribution of preservatives in wood during exposure trials can be assessed by spectral imaging methods although standardized methods are not available. This study concludes that, in addition to the many standard methods available, there is a need to develop new faster methods for screening potential preservative formulation while maintaining the comparability and relevance of results.

Keywords: analytics, methods, preservatives, wood decay

Procedia PDF Downloads 203

Authors: Lekha Logu

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The coordination behavior of the newly synthesized Schiff base ligands, 4-bromo-2-((p-tolyl imino) methyl) phenol obtained by condensing para-toluidine with 5-bromo salicylaldehyde and N-(3,4-dichloro benzylidene)-4-methylbenzenamine obtained by condensing Para-toluidine with 3,4-dichloro benzaldehyde in ethanolic medium has been explored in this current study. The synthesized Schiff’s base ligands were complexed with lanthanide nitrate salts yielding [LnL(NO3)2(H2O)2]NO3, (Ln=Pr, Sm). Elemental analysis, conductance measurement, and spectral techniques like Nuclear Magnetic Resonance (NMR), Ultraviolet-visible (UV-Vis) and Fourier Transform Infrared (FTIR) have been used to characterize Schiff’s base ligands and their lanthanide metal complexes. An attempt has been made on these complexes for their antimicrobial activity against the gram-positive and gram-negative bacterial species like Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Klebsiella pneumonia and fungal species like Canadida and Aspergillus.

Keywords: lanthanide complexes, Schiff's base, antimicrobial assay, synthesis, characterization

Procedia PDF Downloads 42

Authors: Dipali Nagaonkar, Mahendra Rai

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Nanotechnology has experienced significant developments in engineered nanomaterials in the core-shell arrangement. Nanomaterials having nanolayers of silver and gold are of primary interest due to their wide applications in catalytical and biomedical fields. Further, mycosynthesis of nanoparticles has been proved as a sustainable synthetic approach of nanobiotechnology. In this context, we have synthesized silver and silver (core)-gold (shell) bimetallic nanoparticles using a fungal extract of Fusarium graminearum by sequential reduction. The core-shell deposition of nanoparticles was confirmed by the red shift in the surface plasmon resonance from 434 nm to 530 nm with the aid of the UV-Visible spectrophotometer. The mean particle size of Ag and Ag-Au nanoparticles was confirmed by nanoparticle tracking analysis as 37 nm and 50 nm respectively. Quite polydispersed and spherical nanoparticles are evident by TEM analysis. These mycosynthesized bimetallic nanoparticles were tested against some pathogenic bacteria and Candida sp. The antimicrobial analysis confirmed enhanced anticandidal and antibacterial potential of bimetallic nanoparticles over their monometallic counterparts.

Keywords: bimetallic nanoparticles, core-shell arrangement, mycosynthesis, sequential reduction

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Authors: Seyed Reza Aghili, Tahereh Shokohi, Shirin Sadat Hashemi Fesharaki, Mohammad Ali Boroumand, Bahar Salmanian

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Introduction: Intravenous catheter-associated fungal infection as nosocomial infection continue to be a deep problem among hospitalized patients, decreasing quality of life and adding healthcare costs. The capacity of catheters in the spread of candidemia in heart failure patients is obvious. The aim of this study was to evaluate the prevalence and genetic identification of Candida species in heart disorder patients. Material and Methods: This study was conducted in Tehran Hospital of Cardiology Center (Tehran, Iran, 2014) during 1.5 years on the patients hospitalized for at least 7 days and who had central or peripheral vein catheter. Culture of catheters, blood and skin of the location of catheter insertion were applied for detecting Candida colonies in 223 patients. Identification of Candida species was made on the basis of a combination of various phenotypic methods and confirmed by sequencing the ITS1-5.8S-ITS2 region amplified from the genomic DNA using PCR and the NCBI BLAST. Results: Of the 223 patients samples tested, we identified totally 15 Candida isolates obtained from 9 (4.04%) catheter cultures, 3 (1.35%) blood cultures and 2 (0.90%) skin cultures of the catheter insertion areas. On the base of ITS region sequencing, out of nine Candida isolates from catheter, 5(55.6%) C. albicans, 2(22.2%) C. glabrata, 1(11.1%) C. membranifiaciens and 1 (11.1%) C. tropicalis were identified. Among three Candida isolates from blood culture, C. tropicalis, C. carpophila and C. membranifiaciens were identified. Non-candida yeast isolated from one blood culture was Cryptococcus albidus. One case of C. glabrata and one case of Candida albicans were isolated from skin culture of the catheter insertion areas in patients with positive catheter culture. In these patients, ITS region of rDNA sequence showed a similarity between Candida isolated from the skin and catheter. However, the blood samples of these patients were negative for fungal growth. We report two cases of catheter-related candidemia caused by C. membranifiaciens and C. tropicalis on the base of genetic similarity of species isolated from blood and catheter which were treated successfully with intravenous fluconazole and catheter removal. In phenotypic identification methods, we could only identify C. albicans and C. tropicalis and other yeast isolates were diagnosed as Candida sp. Discussion: Although more than 200 species of Candida have been identified, only a few cause diseases in humans. There is some evidence that non-albicans infections are increasing. Many risk factors, including prior antibiotic therapy, use of a central venous catheter, surgery, and parenteral nutrition are considered to be associated with candidemia in hospitalized heart failure patients. Identifying the route of infection in candidemia is difficult. Non-albicans candida as the cause of candidemia is increasing dramatically. By using conventional method, many non-albicans isolates remain unidentified. So, using more sensitive and specific molecular genetic sequencing to clarify the aspects of epidemiology of the unknown candida species infections is essential. The positive blood and catheter cultures for candida isolates and high percentage of similarity of their ITS region of rDNA sequence in these two patients confirmed the diagnosis of intravenous catheter-associated candidemia.

Keywords: catheter-associated infections, heart failure patient, molecular genetic sequencing, ITS region of rDNA, Candidemia

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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

Abstract:

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: A. K. Tursunova, O. V. Chebonenko, A. Zh. Amirkulova, A. O. Abaildayev, O. A. Sapko, Y. M. Dyo, A. Sh. Utarbaeva

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Fusarium solani and its variants cause root and stem rot of plants. Dry rot is the most common disease of potato tubers during storage. The causative agents of fusariosis in contact with plants behave as antagonists, growth stimulants or parasites. The diversity of host-parasite relationships is explained by the parasite’s ability to produce a wide spectrum of biologically active compounds including toxins, enzymes, oligosaccharides, antibiotic substances, enniatins and gibberellins. Many of these metabolites contribute to the creation of compatible relations; others behave as elicitors, inducing various protective responses in plants. An important part of the strategy for developing plant resistance against pathogens is the activation of protein synthesis to produce protective ‘pathogenesis-related’ proteins. The family of PR-proteins known to confer the most protective response is chitinases (EC 3.2.1.14, Cht) and β-1,3-glucanases (EC 3.2.1.39, Glu). PR-proteins also include a large multigene family of peroxidases (EC 1.11.1.7, Pod), and increased activity of Pod and expression of the Pod genes leads to the development of resistance to a broad class of pathogens. Despite intensive research on the role of PR-proteins, the question of their participation in the mechanisms of formation of the F.solani–S.tuberosum pathosуstem is not sufficiently studied. Our aim was to investigate the effect of different classes of F. solani metabolites on the activity of chitinase, β-1,3-glucanases and peroxidases in tubers of Solanum tuberosum. Metabolite culture filtrate (CF) and cytoplasmic components were fractionated by extraction of the mycelium with organic solvents, salting out techniques, dialysis, column chromatography and ultrafiltration. Protein, lipid, carbohydrate and polyphenolic fractions of fungal metabolites were derived. Using enzymatic hydrolysis we obtained oligo glycans from fungal cell walls with different molecular weights. The activity of the metabolites was tested using potato tuber discs (d = 16mm, h = 5mm). The activity of PR-proteins of tubers was analyzed in a time course of 2–24 hours. The involvement of the analysed metabolites in the modulation of both early non-specific and late related to pathogenesis reactions was demonstrated. The most effective inducer was isolated from the CF (fraction of total phenolic compounds including naphtazarins). Induction of PR-activity by this fraction was: chitinase - 340-360%, glucanase - 435-450%, soluble forms of peroxidase - 400-560%, related forms of peroxidase - 215-237%. High-inducing activity was observed by the chloroform and acetonitrile extracts of the mycelium (induction of chitinase and glucanase activity was 176-240%, of soluble and bound forms of peroxidase - 190-400%). The fraction of oligo glycans mycelium cell walls of 1.2 kDa induced chitinase and β-1,3-glucanase to 239-320%; soluble forms and related peroxidase to 198-426%. Oligo glycans cell walls of 5-10 kDa had a weak suppressor effect - chitinase (21-25%) and glucanase (25-28%) activity; had no effect on soluble forms of peroxidase, but induced to 250-270% activity related forms. The CF polysaccharides of 8.5 kDa and 3.1 kDa inhibited synchronously the glucanase and chitinase specific response in step (after 24 hours at 42-50%) and the step response induced nonspecific peroxidase activity: soluble forms 4.8 -5.2 times, associated forms 1.4-1.6 times.

Keywords: fusarium solani, PR-proteins, peroxidase, solanum tuberosum

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Authors: Sami Michael Ayodele

Abstract:

Cordyceps are members of the fungi family Cordycepitaceae which are formally in Clavicipitaceae. They parasitize the larvae and adults of different susceptible arthropods. They are highly priced and have been reported to be highly medicinal. Cordyceps have been used for centuries in traditional medicine in China and other Asian countries. Survey studies were carried out to determine the presence of Cordyceps and another entomopathogenic fungal floral in North Central Nigeria for two consecutive years. Thirty samples were collected, twenty-eight were identified, and two were unidentified. Most identified species belong to Ophiocordyceps species (20samples), Cordyceps species (4samples) and Isaria species (4samples). The species identified were: Cordyceps pseudomilitaris, Cordyceps tuberculata, Cordyceps cylindrical, Ophiocordyceps nutans, O. criminals, O. oxycephala, O. kniphofioides and Isaria sinclairii. The morphological and microscopic features of the collected and identified species were similar to those reported in other countries. This is the first official report on the presence of Cordyceps species in Nigeria. Further collections from different ecological regions of Nigeria will show the richness of the floral diversity of these entomopathogenic and medicinal mushrooms in Nigeria.

Keywords: cordyceps, entomopathogenic, medicinal, North Central, Nigeria

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Authors: Noura Kichou, Manel Tafergguenit, Nabila Ghechtouli, Zakia Hank

Abstract:

The aim of this work is to synthesize, characterize and evaluate the biological activity of two Ligands : glyoxime and dimethylglyoxime, and their metal Ni(II) chelates. The newly chelates were characterized by elemental analysis, IR, EPR, nuclear magnetic resonances (1H and 13C), and biological activity. The antibacterial and antifungal activities of the ligands and its metal complexes were screened against bacterial species (Staphylococcus aureus, Bacillus subtilis, and Escherichia coli) and fungi (Candida albicans). Ampicillin and amphotericin were used as references for antibacterial and antifungal studies. The activity data show that the metal complexes have a promising biological activity comparable with parent free ligand against bacterial and fungal species. A structural, energetic, and electronic theoretical study was carried out using the DFT method, with the functional B3LYP and the gaussian program 09. A complete optimization of geometries was made, followed by a calculation of the frequencies of the normal modes of vibration. The UV spectrum was also interpreted. The theoretical results were compared with the experimental data.

Keywords: glyoxime, dimetylglyoxime, nickel, antibacterial activity

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Authors: Noura Kichou, Manel Tafergguenit, Nabila Ghechtouli, Zakia Hank

Abstract:

The aim of this work is to synthesize, characterize and evaluate the biological activity of two Ligands: glyoxime and dimethylglyoxime, and their metal Ni(II) chelates. The newly chelates were characterized by elemental analysis, IR, EPR, nuclear magnetic resonances (1H and 13C), and biological activity. The antibacterial and antifungal activities of the ligands and its metal complexes were screened against bacterial species (Staphylococcus aureus, Bacillus subtilis, and Escherichia coli) and fungi (Candida albicans). Ampicillin and amphotericin were used as references for antibacterial and antifungal studies. The activity data show that the metal complexes have a promising biological activity comparable with parent free ligand against bacterial and fungal species. A structural, energetic, and electronic theoretical study was carried out using the DFT method, with the functional B3LYP and the gaussian program 09. A complete optimization of geometries was made, followed by a calculation of the frequencies of the normal modes of vibration. The UV spectrum was also interpreted. The theoretical results were compared with the experimental data.

Keywords: glyoxime, dimetylglyoxime, nickel, antibacterial activity

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Authors: Rima Zakzouk, Noriko Hiraishi, Mohamed Mahdi Alshahni, Koichi Makimura, Junji Tagami

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Background and Purpose: Silver diamine fluoride (SDF) is used to prevent and arrest dental caries. The aim of this study is to evaluate the effect of SDF on reducing Candida albicans adhesion on dentin. Materials and Methods: Bovine dentin disks (6×6 mm) were cut by Isomet and polished using grit silicon carbide papers down to 2000 in order to obtain flat dentin surfaces. Samples were divided into two groups. The first group (SDF group) was treated with 38% SDF for 3 min, while the other group (control group) did not undergo SDF treatment. All samples were exposed to C. albicans suspension, washed after 6 hours incubation at 30 °C before to be tested using XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) and real time PCR approaches. Statistical analyses of the results were performed at the significance level α = 0.05. Results: SDF inhibited C. albicans adhesion onto dentin. A significant difference was found between the SDF and control groups in both XTT and real time PCR tests. Conclusion: Using SDF to arrest the caries, could inhibit the Candida growth on dentin.

Keywords: silver diamine fluoride, dentin, real time PCR, XTT

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Authors: Seonggil Kim, Jeonghun Nam, Chae Seung Lim

Abstract:

Rapid diagnosis of fungal infections is critical for rapid antifungal therapy. However, it is difficult to detect extremely low concentration fungi in blood sample. To address the limitation, separation and concentration of fungi in blood sample are required to enhance the sensitivity of PCR analysis. In this study, we demonstrated a sheathless separation and concentration of fungi, candida cells using a viscoelastic fluid. To validate the performance of the device, microparticle mixture (2 and 13 μm) was used, and those particles were successfully separated based on the size difference at high flow rate of 100 μl/min. For the final application, successful separation of the Candida cells from the white blood cells (WBCs) was achieved. Based on the viscoelastic lateral migration toward the equilibrium position, Candida cells were separated and concentrated by center focusing, while WBCs were removed by patterning into two streams between the channel center and the sidewalls. By flow cytometric analysis, the separation efficiency and the purity were evaluated as ~99% and ~ 97%, respectively. From the results, the device can be the powerful tool for detecting extremely rare disease-related cells.

Keywords: candida cells, concentration, separation, viscoelastic fluid

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Authors: Paiwan Buachan, Maneekarn Namsa-Aid, Suchada Jongrungruangchok, Foengchat Jarintanan, Wanlaya Uthaisang-Tanechpongtamb

Abstract:

Lung cancer or pulmonary carcinoma is the uncontrolled growth of abnormal cells in one or both of the lungs. These abnormal cells can spread to other organs of the body through lymphatic system or bloodstream which is called metastatic stage that leading cause of cancer death. Terrein (C₈H₁₀O₃; MW= 154.06 kDa) is a secondary bioactive fungal metabolite, which was isolated from the Aspergillus terreus. In this study, we investigated the effects of terrein on the inhibition of human lung cancer cell proliferation and metastasis. The A549 human non-small cell lung cancer cell line was used as a model. Terrein significantly inhibited lung cancer cell proliferation measuring by a colorimetric MTT assay (IC₅₀ 0.32 mM) and significantly inhibited metastatic processes including migration, invasion, and adhesion that determined by wound healing assay, transwell assay, and adhesion assay, respectively. These findings indicate that terrein could be a potential therapeutic agent for lung cancer.

Keywords: terrein, lung cancer, anticancer, antimetastatic

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Authors: Abdallah Habbab, Khaled Sekkoum, Nasser Belboukhari

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Objective: This study is designed to examine the chemical composition, antioxidant and antibacterial activities of the essential oil extracted from leaves of Vitex agnus-castus. Methods: The essential oils of dry leaves of Vitex agnus-castus L. were obtained by hydro-distillation, afforded oil in the yield of 5.5% and their volatile constituents were identified by GC/MS. Antioxidant activity of the sample was determined by test system DPPH. Antifungal activity was tested against three fungal strains (Aspergillus flavus, Penicillium escpansum and Aspergillus ochraceus) by direct contact method. Results: Forty-three compounds were identified, representing 98.02% of the oil. Major components of the oil were 1,8-cineole (18.27 %), caryophyllene (8.60 %), N-(M-fluorophenyl)-maleimide (6.30 %), (+)-epi-bicyclosesquiphellandrene (6.00 %), terpinen-4-ol (5.57 %), pyrrolo (3,2,1-jk) carbazole (5.43 %), caryophyllene oxide (4.79 %), and phenol (4.09 %). Conclusion: The chemical constituents in the essential oil from the locally grown Vitex agnus-castus were identified. Therefore, the essential oil of Vitex agnus-castus is an active candidate which would be used as antioxidant, or antifungal agent in new drugs preparation for therapy of diseases.

Keywords: Vitex agnus-castus, essential oil, GC/MS, DPPH, 1, 8-cineole

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Authors: Jessica P. Fiorotti, Maria C. Freitas, Caio J. B. Coutinho-Rodrigues, Mariana G. Camargo, Emily S. Mesquita, Amanda R. C. Corval, Ricardo O. B. Bitencourt, Allan F. Marciano, Diva D. Spadacci-Morena, Patricia S. Golo, Isabele C. Angelo, Vania R. E. P. Bittencourt

Abstract:

The bovine tick, Rhipicephalus microplus, is an arthropod of great importance in veterinary medicine leading to anemia, weight loss, animals' leather depreciation and also acting as a vector of many pathogens. In this way, the parasitism causes a loss of 3.24 billion dollars per year in Brazil. Knowingly, entomopathogenic fungi act as natural controller of some arthropods, acting mainly by active penetration through the cuticle. However, it can also act on the hemolymph and through the production of mycotoxins. Hemocytes are responsible for the cellular immune response and participate in the processes of phagocytosis, nodulation and encapsulation and may undergo changes when challenged by pathogens. The aim of the present study was to evaluate changes in R. microplus hemocytes after inoculation of Metarhizium robertsii using transmission electron microscopy. The isolate ARSEF 2575 and 200 engorged R. microplus females were used. The groups were divided into control, in which the females were inoculated with 5 μL of sterile distilled water solution and 0.1% Tween 80, and a group inoculated with 5 μL of fungal suspension at the concentration of 10⁷ conidia mL⁻¹. The experiment was performed in duplicate and each group contained 50 females. Twenty-four hours after fungal inoculation, hemolymph was collected through the cuticle dorsal surface perforation of the tick females. After collection, the hemolymph samples were centrifuged at 500 x g for 3 minutes at 4 °C, the plasma was discarded and the hemocyte pellet was resuspended in 50 μl PBS. The suspension material was fixed in 2% glutaraldehyde in Millonig buffer for three hours. After fixation, the material was centrifuged at 500 x g for 3 minutes, the supernatant was discarded and the cells were resuspended in a wash solution. Subsequently, the cells were post-fixed with 1% osmium tetroxide in phosphate buffer for one hour at room temperature and dehydrated in increasing concentrations of ethanol, and then embedded in Epon resin. The ultrathin sections were examined under the LEO EM 906E transmission electron microscopy at 80kV. The ultrastructural results revealed that.in control group, the cells were considered intact, in which the granulocytes were observed with granules of different electrodensities, intact mitochondria and cytoplasm without vacuolization. In addition, granulocytes showed plasma membrane projections similar to pseudopodia. Plasmatocytes presented as irregularly shaped cells, with the eccentric nucleus, agranular cytoplasm and some cells presented pseudopodia. Nevertheless, in the group exposed to the fungus, most of the cells presented in degeneration. The granulocytes found had fewer granules in the cytoplasm and more vacuoles. Plasmatocytes, after treatment, presented many vacuoles also in the cytoplasm and the lysosomes presented great amount of electrodense material in their interior. Thus, the results suggest that the fungus has a depressant action in the immune system of the tick, not only by the cell degranulation, but also suggesting that this leads to morphological changes in the hemocytes and may even trigger processes such as phagocytosis.

Keywords: bovine tick, cellular defense, entomopathogenic fungi, immune response

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