Search results for: entomopathogenic fungi silkworm larva

Authors: Ahmed M. Haddad, Hadeel S. El-Shaal, Gadallah M. Abu-Elreesh

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Some of filamentous fungi can be used for biodiesel production as they are able to accumulate high amounts of intracellular lipids when grown at stress conditions. Aspergillus fumigatus sp. was isolated from Nile delta soil in Egypt. The fungus was primarily screened for its capacity to accumulate lipids using Nile red staining assay. The fungus could accumulate more than 20% of its biomass as lipids when grown at optimized minimal medium. After lipid extraction, we could use fungal cell debris to remove some heavy metals from contaminated waste water. The fungal cell debris could remove Cd, Cr, and Zn with absorption efficiency of 73%, 83.43%, and 69.39% respectively. In conclusion, the Aspergillus fumigatus isolate may be considered as a promising biodiesel producer, and its biomass waste can be further used for bioremediation of wastewater contaminated with heavy metals.

Keywords: biodiesel, bioremediation, fungi, heavy metals, lipids, oleaginous

Procedia PDF Downloads 194

Authors: Elin Julianti, Marlia Singgih, Masayoshi Arai, Jianyu Lin, Masteria Yunovilsa Putra, Muhammad Azhari, Agnia S. Muharam

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The search for a source of new medicinal compounds with anticancer activity from natural products has become important to resolve the ineffectiveness problem of pancreatic cancer therapy. Fungal marine microorganisms are prolific sources of bioactive natural products. In this present study, the ethyl acetate extract of cultured broth of Trichoderma longibrachiatum marine sponge-derived fungi exhibited selective cytotoxicity against human pancreatic carcinoma PANC-1 cells cultured under glucose-deficient conditions (IC50 = 98,4 µg/mL). The T. longibrachiatum was fermented by the static method at room temperature for 60 days. The culture broth was extracted using ethyl acetate by liquid-liquid extraction method. The liquid-liquid extraction was conducted toward the ethyl extract by using 90% MeOH-H₂O and n-|Hexane as a solvent. The extract of 90% MeOH-H₂O was fractionated by liquid extraction using by C₁₈ reversed-phase vacuum flash chromatography using mixtures of MeOH-H₂O, from 50:50 to 100:0, and 1% TFA MeOH as the eluents to yield six fractions. The fraction 2 (MeOH-H2O, 70:30) and fraction 3 (MeOH-H2O, 80:20) showed moderate cytotoxicity with IC50 value of 119.3 and 274.7 µg/mL, respectively. Fraction 4 (MeOH-H₂O, 90:10) showed the highest cytotoxicity activity with IC₅₀value of < 10 µg/mL. The chemical compounds of the fractions that are responsible for cytotoxic activity are potent for further investigation.

Keywords: cytotoxic activity, trichoderma longibrachiatum, marine-derived fungi, PANC-1 cell line

Procedia PDF Downloads 258

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 316

Authors: Hai-Hong Gu, Yan-Jun Ai, Zheng Zhou

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Vanadium and titanium are rare metals with superior properties and are important resources in aerospace, aviation, and military. The vanadium/titanium magnetite are mostly ultra-lean ores, and a large number of tailings has been produced in the exploitation process. The tailings are characterized by loose structure, poor nutrient, complex composition and high trace metal contents. Returning green manure has been shown to not only increase plant biomass and soil nutrients but also change the bioavailability of trace metals and the microbial community structure. Fungi play an important role in decomposing organic matter and increasing soil fertility, and the application of organic matter also affects the community structure of fungi. The effects of green manure plants, alfalfa (Medicago sativa L.), returned to the tailings in situ on community structure of fungi, nutrients and bioavailability of trace metals in vanadium/titanium magnetite tailings were investigated in a pot experiment. The results showed that the fungal community diversity and richness were increase after alfalfa green manure returned in situ. The dominant phyla of the fungal community were Ascomycota, Basidiomycota and Ciliophora, especially, the phyla Ciliophora was rare in ordinary soil, but had been found to be the dominant phyla in tailings. Meanwhile, the nutrient properties and various trace metals may shape the microbial communities by affecting the abundance of fungi. It was found that the plant growth was stimulated and the available N and organic C were significantly improved in the vanadium/titanium magnetite tailing with the long-term returning of alfalfa green manure. Moreover, the DTPA-TEA extractable Cd and Zn concentrations in the vanadium/titanium magnetite tailing were reduced by 7.72%~23.8% and 8.02%~24.4%, respectively, compared with those in the non-returning treatment. The above results suggest that the returning of alfalfa green manure could be a potential approach to improve fungal community structure and restore mine tailing ecosystem.

Keywords: fungal community, green manure returning, vanadium/titanium magnet tailings, trace metals

Procedia PDF Downloads 37

Authors: Shivankar Agrawal, Sunil Kumar Deshmukh, Colin Barrow, Alok Adholeya

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A variety of genetic and environmental factors cause various cosmetics and dermatological problems. There are already claimed drugs available in market for treating these problems. However, the challenge remains in finding more potent, environmental friendly, causing minimal side effects and economical cosmeceuticals. This leads to an increased demand for natural cosmeceutical products in the last few decades. Plant derived ingredients are limited because plants either contain toxic metabolites, grow too slow or seasonal harvesting is a problem. To identify new bioactive cosmetics ingredients of marine microbial bioresource, we screened 35 marine fungi isolated from marine samples collected from Andaman Island and west coast of India. Fungal crude extracts were investigated for their antityrosinase, antioxidant and antibacterial activities for the purpose of identifying anti-aging, skin-whitening and anti-acne biomolecule with the potential in cosmetics. In the tyrosinase inhibition and 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assays, two fungal extracts, including “P2”, Talaromyces stipitatus and “D4”, Aspergillus terreus showed high inhibitory activity at 1mg/mL for tyrosinase inhibition and 0.5mg/mL for DPPH scavenging. The in vitro antimicrobial activity was investigated by the agar well diffusion method. In the tyrosinase inhibition assay, 8 extracts showed significant antibacterial activity against bacteria causing skin and wound infection in humans. In the course of systematic screening program for bioactive marine fungi, strain “D5” was found to be most potent strain with MIC value of 1mg/mL, which was morphologically identified as Simplicillium lamellicola. The effects of the most active crude extracts against their susceptible test microorganisms were also investigated by SEM analysis. Further investigations will focus on purification and characterization major active components responsible for these activities.

Keywords: antioxidant, antimicrobial activity, tyrosinase, cosmeceuticals, marine fungi

Procedia PDF Downloads 256

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 223

Authors: Sean T. S. Wei, Joy D. Van Nostrand, Annapoorna Maitrayee Ganeshram, Stephen B. Pointing

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McMurdo Dry Valleys are a largely ice-free polar desert protected by international treaty as an Antarctic special managed area. The terrestrial landscape is dominated by oligotrophic mineral soil with extensive rocky outcrops. Several environmental stresses: low temperature, lack of liquid water, UV exposure and oligotrophic substrates, restrict the major biotic component to microorganisms. The bacterial diversity and the putative physiological capacity of microbial communities of quartz rocks (hypoliths) and soil of a maritime-influenced Dry Valleys were interrogated by two metagenomic approaches: 454 pyro-sequencing and Geochp DNA microarray. The most abundant phylum in hypoliths was Cyanobacteria (46%), whereas in solils Actinobacteria (31%) were most abundant. The Proteobacteria and Bacteriodetes were the only other phyla to comprise >10% of both communities. Carbon fixation was indicated by photoautotrophic and chemoautotrophic pathways for both hypolith and soil communities. The fungi accounted for polymer carbon transformations, particularly for aromatic compounds. The complete nitrogen cycling was observed in both communities. The fungi in particular displayed pathways related to ammonification. Environmental stress response pathways were common among bacteria, whereas the nutrient stress response pathways were more widely present in bacteria, archaea and fungi. The diversity of bacterialphage was also surveyed by Geochip. Data suggested that different substrates supported different viral families: Leviviridae, Myoviridae, Podoviridae and Siphoviridiae were ubiquitous. However, Corticoviridae and Microviridae only occurred in wetter soils.

Keywords: Antarctica, hypolith, soil, dry valleys, geochip, functional diversity, stress response

Procedia PDF Downloads 421

Authors: Mohammad Abdollahi

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Tomato (Lycopersicon esculentum Mill.) is one of the most popular, widely grown and the second most important vegetable crop, after potatoes. Nematodes have been identified as one of the major pests affecting tomato production throughout the world. The most destructive nematodes are the genus Meloidogyne. Most widespread and devastating species of this genus are M. incognita, M. javanica, and M. arenaria. These species can cause complete crop loss under adverse growing conditions. There are several potential methods for management of the root knot nematodes. Although the chemicals are widely used against the phytonematodes, because of hazardous effects of these compounds on non-target organisms and on the environment, there is a need to develop other control strategies. Nowadays, non-chemical measures are widely used to control the plant parasitic nematodes. Biocontrol of phytonematodes is an important method among environment-friendly measures of nematode management. There are some soil-inhabiting fungi that have biocontrol potential on phytonematodes, which can be used in nematode management program. The fungus Metarhizium anisopliae, originally is an entomopathogenic bioagent. Biocontrol potential of this fungus on some phytonematodes has been reported earlier. Recently, use of organic soil amendments as well as the use of bioagents is under special attention in sustainable agriculture. This research aimed to reduce the pesticide use in control of root-knot nematode, Meloidogyne javanica in tomato. The effects of M. anisopliae IMI 330189 and different levels of oak tree debris on M. javanica were determined. The combination effect of the fungus as well as the different rates of soil amendments was determined. Pots were filled with steam pasteurized soil mixture and the six leaf tomato seedlings were inoculated with 3000 second stage larvae of M. javanica/kg of soil. After eight weeks, plant growth parameters and nematode reproduction factors were compared. Based on the results of our experiment, combination of M. anisopliae IMI 330189 and oak debris caused more than 90% reduction in reproduction factor of nematode, at the rates of 100 and 150 g/kg soil (P ≤ 0.05). As compared to control, the reduction in number of galls was 76%. It was 86% for nematode reproduction factor, showing the significance of combined effect of both tested agents. Our results showed that plant debris can increase the biological activity of the tested bioagent. It was also proved that there was no adverse effect of oak debris, which potentially has antimicrobial activity, on antagonistic power of applied bioagent.

Keywords: biological control, nematode management, organic soil, Quercus branti, root knot nematode, soil amendment

Procedia PDF Downloads 141

Authors: Aya Tanaka, Mariko Era, Manami Masuda, Yui Okuno, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives— Fungi and bacteria are present in a wide range of natural environments. They are breed in the foods such as vegetables and fruit, causing corruption and deterioration of these foods in some cases. Furthermore, some species of fungi and bacteria are known to cause food intoxication or allergic reactions in some individuals. To prevent fungal and bacterial contamination, various fungicides and bactericidal have been developed that inhibit fungal and bacterial growth. Fungicides and bactericides must show high antifungal and antibacterial activity, sustainable activity, and a high degree of safety. Therefore, we focused on the fatty acid salt which is the main component of soap. We focused on especially C10K and C12K. This study aimed to find the effectiveness of the fatty acid salt as antimicrobial agents for food safety. Materials and Methods— Cladosporium cladosporioides NBRC 30314, Penicillium pinophilum NBRC 6345, Aspergillus oryzae (Akita Konno store), Rhizopus oryzae NBRC 4716, Fusarium oxysporum NBRC 31631, Escherichia coli NBRC 3972, Bacillus subtilis NBRC 3335, Staphylococcus aureus NBRC 12732, Pseudomonas aenuginosa NBRC 13275 and Serratia marcescens NBRC 102204 were chosen as tested fungi and bacteria. Hartmannella vermiformis NBRC 50599 and Acanthamoeba castellanii NBRC 30010 were chosen as tested amoeba. Nine fatty acid salts including potassium caprate (C10K) and laurate (C12K) at 350 mM and pH 10.5 were used as antifungal activity. The spore suspension of each fungus (3.0×10⁴ spores/mL) or the bacterial suspension (3.0×10⁵ or 3.0×10⁶ or 3.0×10⁷ CFU/mL) was mixed with each of the fatty acid salts (final concentration of 175 mM). Samples were counted at 0, 10, 60, and 180 min by plating (100 µL) on potato dextrose agar or nutrient agar. Fungal and bacterial colonies were counted after incubation for 1 or 2 days at 30 °C. Results— C10K was antifungal activity of 4 log-unit incubated time for 10 min against fungi other than A. oryzae. C12K was antifungal activity of 4 log-unit incubated time for 10 min against fungi other than P. pinophilum and A. oryzae. C10K and C12K did not show high anti-yeast activity. C10K was antibacterial activity of 6 or 7 log-unit incubated time for 10 min against bacteria other than B. subtilis. C12K was antibacterial activity of 5 to 7 log-unit incubated time for 10 min against bacteria other than S. marcescens. C12K was anti-amoeba activity of 4 log-unit incubated time for 10 min against H. vermiformis. These results suggest C10K and C12K have potential in the field of food safety.

Keywords: food safety, microbes, antimicrobial, fatty acid salts

Procedia PDF Downloads 457

Authors: Shameel Pervez, Saad Aziz Durrani, Ibatsam Khokhar

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Pectinase is an enzyme that breaks down pectin, a compound responsible for structural integrity of the plant. Pectin is difficult to break down mechanically and the cost is very high, that is why many industries including food industries use pectinase enzyme produced by microbes for pectin breakdown. Apple (Malus domestica) is an important fruit in terms of market value. Every year, millions of apples are wasted due to post-harvest rot caused by fungi. Fungi are natural decomposers of our ecosystem and are infamous for post-harvest rot of apple fruit but at the same time they are prized for their high production of valuable extracellular enzymes such as pectinase. In this study, fungi belonging to different genus were isolated from rotten apples. Rotten samples of apple were picked from different markets of Lahore. After surface sterilization, the rotten parts were cut into small pieces and placed onto MEA media plates for three days. Afterwards, distinct colonies were picked and purified by sub-culturing. The isolates were identified to genus level through the study of basic colony morphology and microscopic features. The isolates were then subjected to screening for pectinase activity on MS media to compare pectinase production and were then subsequently tested for pathogenic activity through wound suspension method to evaluate the pathogenic activity of isolates in comparison with their pectinolytic activity. A total of twelve fungal strains were isolates from rotten apples. They were belonging to genus Penicillium, Alternaria, Paecilomyces and Rhizopus. Upon screening for pectinolytic activity, isolates Pen 1, Pen 4, and Rz showed high pectinolytic activity and were further subjected to DNA isolation and partial sequencing for species identification. The results of partial sequencing were combined with in-depth study of morphological features revealing Pen 1 as Penicillium janthinellum, Pen 4 as Penicillium griseofulvum, and Rz as Rhizopus microsporus. Pathogenic activity of all twelve isolates was evaluated. Penicillium spp. were highly pathogenic and destructive and same was the case with Paecilomyces sp. and Rhizopus sp. However, Alternaria spp. were found to be more consistent in their pathogenic activity, on all types of apples.

Keywords: apple, pectinase, fungal pathogens, penicillium, rhizopus

Procedia PDF Downloads 28

Authors: Minh-Phuong Ngoc Bui, Abdennour Abbas

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Microbial spoilage of food/drug has been a constant nuisance and an unavoidable problem throughout history that affects food/drug quality and safety in a variety of ways. A simple and rapid test of fungi and bacteria in food/drugs and environmental clinical samples is essential for proper management of contamination. A number of different techniques have been developed for detection and enumeration of foodborne microorganism including plate counting, enzyme-linked immunosorbent assay (ELISA), polymer chain reaction (PCR), nucleic acid sensor, electrical and microscopy methods. However, the significant drawbacks of these techniques are highly demand of operation skills and the time and cost involved. In this report, we introduce a rapid method for detection of bacteria and fungi in food/drug products using a specific interaction between a reducing agent (tris(2-carboxylethyl)phosphine (TCEP)) and the microbial surface proteins. The chemical reaction was transferred to a transduction system using gold nanoplates-enhanced chemiluminescence. We have optimized our nanoplates synthetic conditions, characterized the chemiluminescence parameters and optimized conditions for the microbial assay. The new detection method was applied for rapid detection of bacteria (E.coli sp. and Lactobacillus sp.) and fungi (Mucor sp.), with limit of detection as low as single digit cells per mL within 10 min using a portable luminometer. We expect our simple and rapid detection method to be a powerful alternative to the conventional plate counting and immunoassay methods for rapid screening of microorganisms in food/drug products.

Keywords: microorganism testing, gold nanoplates, chemiluminescence, reducing agents, luminol

Procedia PDF Downloads 269

Authors: Samuel E. Okere, Anthony E. Ataga

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Introduction: The microbial ecology of Pleurotus osteratus and Pleurotus tuber–regium spent mushroom substrate (SMS) were characterized to determine other ways of its utilization. Materials and Methods: The microbiological properties of the spent mushroom substrate were determined using standard methods. This study was carried out at the Microbiology Laboratory University of Port Harcourt, Rivers State, Nigeria. Results: Quantitative microbiological analysis revealed that Pleurotus osteratus spent mushroom substrate (POSMS) contained 7.9x10⁵ and 1.2 x10³ cfu/g of total heterotrophic bacteria and total fungi count respectively while Pleurotus tuber-regium spent mushroom substrate (PTSMS) contained 1.38x10⁶ and 9.0 x10² cfu/g of total heterotrophic bacteria count and total fungi count respectively. The fungi species encountered from Pleurotus tuber-regium spent mushroom substrate (PTSMS) include Aspergillus and Cladosporum species, while Aspergillus and Penicillium species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). However, the bacteria species encountered from Pleurotus tuber-regium spent mushroom substrate include Bacillus, Acinetobacter, Alcaligenes, Actinobacter, and Pseudomonas species while Bacillus, Actinobacteria, Aeromonas, Lactobacillus and Aerococcus species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). Conclusion: Therefore based on the findings from this study, it can be concluded that spent mushroom substrate contain microorganisms that can be utilized both in bioremediation of oil-polluted soils as they contain important hydrocarbon utilizing microorganisms such as Penicillium, Aspergillus and Bacillus species and also as sources of plant growth-promoting rhizobacteria (PGPR) such as Pseudomonas and Bacillus species which can induce resistance on plants. However, further studies are recommended, especially to molecularly characterize these microorganisms.

Keywords: characterization, microorganisms, mushroom, spent substrate

Procedia PDF Downloads 126

Authors: Gabriel Roveda-Hoyos, Margarita Ramirez-Gomez, Adrian Perez, Diana Paola Serralde

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The objective of this study was to determine the variability of arbuscular mycorrhizal fungi (HFMA) communities associated with wild agraz plants (Vaccinium meridionale Swartz) in the Colombian Andes. This species is one of the most promising fruits within the genus Vaccinium because of the high content of anthocyanins and antioxidants in its fruits, and like other species of the Ericaceae family, it depends on the association with HFM for its development in the natural environment. In this study, the presence of mycorrhizae in wild communities of V. meridionale was evaluated, and their relationship with the edaphic and climatic conditions of the study area was analyzed. Sampling was conducted in the rural area of the municipalities of Raquira, and Chiquinquira, Chia, and Tabio in the departments of Cundinamarca and Boyaca, Colombia. Seven sites were selected, and in each site, 5 plants were randomly selected, root and soil samples were taken from each plant in the rhizosphere zone for the quantification of colonization and the presence of spores. The samples were collected on different soils, taxonomic orders Entisols, Inceptisols, and Alfisols, located at altitudes between 2,600 and 3,000 above sea level in the Eastern Cordillera of Colombia. The physicochemical characteristics of the soil were compared with the density of spores and the percentage of presence of mycorrhizae in the roots and variables with the morphometric and physiological characteristics of the plants. Four types of mutual associations were found: arbuscular mycorrhizae, ectendomycorrhiza, ericoid mycorrhizae, and endophytic septate fungi. The main results obtained show a predominance of spores of the genera Glomus and Acaulsopora, in most of the soils analyzed. The spore density of Glomeromycete fungi in the soil varied considerably between the different sites; it was higher ( > 50 spores/g of dry soil) in soil samples with lower bulk density and higher content of organic matter; in these soils a higher cation exchange capacity was found, as well as of nitrogen, calcium, magnesium, manganese and zinc concentration. It can be concluded that Vaccinium meridionale is able to establish in a natural way, association with HFMA.

Keywords: Ericaceae, Arbuscular mycorrhizae, Andes, soils, Glomus sp.

Procedia PDF Downloads 148

Authors: C. Girometta, S. Babbini, R. M. Baiguera, D. S. Branciforti, M. Cartabia, D. Dondi, M. Pellegrini, A. M. Picco, E. Savino

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Interest on wood decay fungi (WDF) has been increasing in the last year's thanks to the potentiality of this kind of fungi; research on new WDF strains has increased as well thus pointing out the key role of the culture collections. One of the most recent biotechnological application of WDF is the development of novel materials from natural or recycled resources. Based on different combinations of fungal species, substrate, and processing treatment involved (e.g. heat pressing), it is possible to achieve a wide variety of materials with different features useful for many industrial applications: from packaging to thermal and acoustic insulation. In comparison with the conventional ones, these materials represent a 100% natural and compostable alternative involving low amounts of energy in the production process. The purpose of the present work was to isolate and select WDF strains able to colonize and degrade different plant wastes thus producing a fungal biomass shapeable to achieve bio-composite materials. Strains were selected within the mycological culture collection of Pavia University (MicUNIPV, over 300 strains of WDF). The selected strains have been investigated with regards their ability to colonize and degrade plant residues from the local major cultivations (e.g. poplar, alfalfa, maize, rice, and wheat) and produce the fungal biomass. The degradation of the substrate was assessed by Thermogravimetric analysis (TGA) and Fourier Transform Infrared Spectroscopy (FTIR). Chemical characterization confirmed that TGA and FTIR are complementary techniques able to provide quality-quantitative information on compositional and structural variation that occurs during the transformation from the substrate to the bio-composite material. This pilot study provides a fundamental step to tune further applications in fungus-residues composite biomaterials.

Keywords: bio-composite material, lignocellulosic residues, sustainable materials, wood decay fungi

Procedia PDF Downloads 110

Authors: Gloria M. Ntuba-Jua, Afui M. Mih, Eneke E. T. Bechem

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Prunus africana (Hook. F.) Kalkman, commonly known as Pygeum or African cherry belongs to the Rosaceae family. It is a medium to large, evergreen tree with a spreading crown of 10 to 20 m. It is used by the traditional medical practitioners for the treatment of over 45ailments in Cameroon and sub-Sahara Africa. In modern medicine, it is used in the treatment of benign prostrate hyperplasia (BPH), prostate gland hypertrophy (enlarged prostate glands). This is possible because of its ability to produce some secondary metabolites which are believed to have bioactivity against these ailments. The ready international market for the sale of Prunus bark, uncontrolled exploitation, illegal harvesting using inappropriate techniques and poor timing of harvesting have contributed enormously to making the plant endangered. It is known to harbor a large number of endophytic fungi with the potential to produce similar secondary metabolites as the parent plant. Alternative sourcing of medicinal principles through endophytic fungi requires succinct knowledge of the endophytic fungi. This will serve as a conservation measure for Prunus africana by reducing dependence on Prunus bark for such metabolites. This work thus sought to compare the production of some major secondary metabolites produced by P. africana and some of its associated endophytic fungi. The leaves and stem bark of the plant from different provenances were soaked in methanol for 72 hrs to yield the methanolic crude extract. The phytochemical screening of the methanolic crude extracts using different standard procedures revealed the presence of tannins, flavonoids, terpenoids, saponins, phenolics and steroids. Pure cultures of some predominantly isolated endophyte species from the difference Prunus provenances such as Curvularia sp, and Morphospecies P001 were also grown in Potato Dextrose Broth (PDB) for 21 days and later extracted with Methylene dichloride (MDC) solvent after 24hrs to produce crude culture extracts. Qualitative assessment of crude culture extracts showed the presence of tannins, terpenoids, phenolics and steroids particularly β-Sitosterol, (a major bioactive metabolite) as did the plant tissues. Qualitative analysis by thin layer chromatography (TLC) was done to confirm and compare the production of β-Sitosterol (as marker compounds) in the crude extracts of the plant and endophyte. Samples were loaded on TLC silica gel aluminium barked plate (Kieselgel 60 F254, 0.2 mm, Merck) using acetone/hexane, (3.0:7.0) solvent system. They were visualized under an ultra violet lamp (UV254 and UV360). TLC revealed that leaves had a higher concentration of β-sitosterol in terms of band intensity than stem barks from the different provenances. The intensity of β-sitosterol bands in the culture extracts of endophytes was comparable to the plant extracts except for Curvularia sp (very minute) whose band was very faint. The ability of these fungi to make β-sitosterol was confirmed by TLC analysis with the compound having chromatographic properties (retention factor) similar to those of β-sitosterol standard. The ability of these major endophytes to produce secondary metabolites similar to the host has therefore been demonstrated. There is, therefore, the potential of developing the in vitro production system of Prunus secondary metabolites thereby enhancing its conservation.

Keywords: Caneroon, endophytic fungi, Prunus africana, secondary metabolite

Procedia PDF Downloads 192

Authors: M. Mehani, N. Salhi, T. Valeria, S. Ladjel

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Red River Gum (Eucalyptus camaldulensis) is a tree of the genus Eucalyptus widely distributed in Algeria and in the world. The value of its aromatic secondary metabolites offers new perspectives in the pharmaceutical industry. This strategy can contribute to the sustainable development of our country. Preliminary tests performed on the essential oil of Eucalyptus camendulensis showed that this oil has antibacterial activity vis-à-vis the bacterial strains (Enterococcus feacalis, Enterobacter cloaceai, Proteus microsilis, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa) and antifungic (Fusarium sporotrichioide and Fusarium graminearum). The culture medium used was nutrient broth Muller Hinton. The interaction between the bacteria and the essential oil is expressed by a zone of inhibition with diameters of MIC indirectly expression of. And we used the PDA medium to determine the fungal activity. The extraction of the aromatic fraction (essentially oil- hydrolat) of the fresh aerian part of the Eucalyptus camendulensis was performed by hydrodistillation. The average essential oil yield is 0.99%. The antimicrobial and fungal study of the essential oil and hydrosol showed a high inhibitory effect on the growth of pathogens.

Keywords: essential oil, Eucalyptus camendulensis, bacteria and fungi, red river gum

Procedia PDF Downloads 202

Authors: Lyudmila V. Ignatova, Yelena V. Brazhnikova, Togzhan D. Mukasheva, Ramza Zh. Berzhanova, Anel A. Omirbekova

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Endophytic microorganisms are presented in plants of different families growing in the foothills and piedmont plains of Trans-Ili Alatau. It was found that the maximum number of endophytic micromycetes is typical to the Fabaceae family. The number of microscopic fungi in the roots reached (145.9±5.9)×103 CFU/g of plant tissue; yeasts - (79.8±3.5)×102 CFU/g of plant tissue. Basically, endophytic microscopic fungi are typical for underground parts of plants. In contrast, yeasts more infected aboveground parts of plants. Small amount of micromycetes is typical to inflorescence and fruits. Antagonistic activity of selected micromycetes against Fusarium graminearum, Cladosporium sp., Phytophtora infestans and Botrytis cinerea phytopathogens was detected. Strains with a broad, narrow and limited range of action were identified. For further investigations Rh2 and T7 strains were selected, they are characterized by a broad spectrum of fungicidal activity and they formed the large inhibition zones against phytopathogens. Active antagonists are attributed to the Rhodotorula mucilaginosa and Beauveria bassiana species.

Keywords: endophytic micromycetes, fungicidal activity, prevalence, plants

Procedia PDF Downloads 294

Authors: S. Zenia, L. Bitta, O. Bouhamam, H. Brines, M. Boudriaa, F. Haddadj, F. Marniche, A. Milla, H. Saadi, A. Smai

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The bee Apis mellifera intermissa is a major social insect, in addition to its honey production, it is a pillar of our biodiversity. Several living organisms can come into contact with it: bacteria, viruses, protozoa, fungi, mites, and insects. In Algeria, many beekeepers have reported unusual mortality of local bees, loss of foragers and significant losses of their livestock. Despite the presence of a varied honey-bearing flora and a favourable Mediterranean climate, honey production remains low. This phenomenon can be attributed to the excess winter mortality, but also to the increasing difficulties that beekeepers face in maintaining healthy bee colonies, particularly bee diseases and their transmission facilitated by trade and beekeeping practices. Our survey is based on a questionnaire composed of several parts. The results obtained show that the disease that most affects bees according to beekeepers is varroa mite with 93% followed by fungi with 26%. The most replied enemy of bees is the false ringworm with 73%, followed by the bee-eater with 63%. Our goal is to determine the causes of this low production in two areas: Bejaia and Tizi-Ouzou.

Keywords: diseases, Apis mellifera L., varroa, European foulbrood

Procedia PDF Downloads 136

Authors: Eusèbe Gnonlonfoun, Xavier Framboisier, Michel Fick, Emmanuel Rondags

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Pathogenic fungi represent a generic problem for cereals, including barley, as they can produce a number of thermostable toxic metabolites such as mycotoxins that contaminate plants and food products, leading to serious health issues for humans and animals and causing significant losses in global food production. In addition, mycotoxins represent a significant technological concern for the malting and brewing industries, as they may affect the quality and safety of raw materials (barley and malt) and final products (beer). Moreover, this situation is worsening due to the highly variable climatic conditions that favor microbial development and the societal desire to reduce the use of phytosanitary products, including fungicides. In this complex environmental, regulatory and economic context for the French barley-malt-beer industry, this project aims to develop an innovative biocontrol process by using technological bacteria, isolated from infection-resistant barley cultures, that are able to reduce the development of spoilage fungi and the associated mycotoxin production. The experimental approach consists of i) coculturing bacterial and pathogenic fungal strains in solid and liquid media to access the growth kinetics of these microorganisms and to evaluate the impact of these bacteria on fungal growth and mycotoxin production; then ii) the results will be used to carry out a micro-malting process in order to develop the aforementioned process, and iii) the technological and sanitary properties of the generated barley malts will finally be evaluated in order to validate the biocontrol process developed. The process is expected to make it possible to guarantee, with controlled costs, an irreproachable hygienic and technological quality of the malt, despite the increasingly complex and variable conditions for barley production. Thus, the results will not only make it possible to maintain the dominant world position of the French barley-malt chain but will also allow it to conquer emerging markets, mainly in Africa and Asia. The use of this process will also contribute to the reduction of the use of phytosanitary products in the field for barley production while reducing the level of contamination of malting plant effluents. Its environmental impact would therefore be significant, especially considering that barley is the fourth most-produced cereal in the world.

Keywords: barley, pathogenic fungi, mycotoxins, malting, bacterial biocontrol

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Authors: Mosaab A. Omar, Mohammad Saleh Al-Aboody

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In the present study, examination of 442 faecal samples was performed: 171 from cattle, 128 from buffaloes and 143 from sheep. During the period from May, 2014 to April, 2015, fecal examination showed the infection rate with abomasal nematodes was 30% in cattle, 22.6% in buffaloes, and 31.4% in sheep. Fecal culture gave results of 47.5%, 30%, and 50.3% in cattle, buffaloes and sheep respectively. Seasonal infection with abomasal nematodes as shown by faecal culture in cattle, reveals the highest infection rate is in summer (55.9%), followed by spring (54.1%), autumn (50%), and winter (33.3%). Cooperia spp. is the most prevalent larva in both cattle and buffaloes; Strongyloides papillosus is the most predominant one in sheep. Here we introduce the first study of abomasal worms infection in ruminants in Qena, Egypt. The prevalence is found to be so high among the all examined animals, that we recommend that the authorities apply suitable control programs.

Keywords: haemonchus, ostertagia, seasonal dynamics, floatation

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Authors: Zhanara Suleimenova, Raushan Blieva, Aigerim Zhakipbekova, Inkar Tapenbayeva, Zhanar Narmuratova

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Green processes are based on innovative technologies that do not negatively affect the environment. Industrial enzymes originated from biological systems can effectively contribute to sustainable development through being isolated from microorganisms which are fermented using primarily renewable resources. Many widespread microorganisms secrete a significant amount of biocatalysts into the environment, which greatly facilitates the task of their isolation and purification. The ability to control the enzyme production through the regulation of their biosynthesis and the selection of nutrient media and cultivation conditions allows not only to increase the yield of enzymes but also to obtain enzymes with certain properties. In this regard, large potentialities are embedded in immobilized cells. Enzyme production technology in a secreted active form enabling industrial application on an economically feasible scale has been developed. This method is based on the immobilization of enzyme producers on a solid career. Immobilizing has a range of advantages: decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, the ability of various enzymes' simultaneous production, etc. Design of proposed equipment gives the opportunity to increase the activity of immobilized cell culture filtrate comparing to free cells, growing in periodic culture conditions. Such technology allows giving a 10-times raise in culture productivity, to prolong the process of fungi cultivation and periods of active culture liquid generation. Also, it gives the way to improve the quality of filtrates (to make them more clear) and exclude time-consuming processes of recharging fermentative vials, that require manual removing of mycelium.

Keywords: industrial enzymes, immobilization, submerged fermentation, microscopic fungi

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Authors: Sehrish Iftikhar, Ahmad Ali Shahid, Kiran Nawaz, Waheed Anwar

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Discovery and development of new compounds require intense studies in chemistry, biochemistry. Numerous experiments under laboratory-, greenhouse- and field conditions can be performed to select suitable candidates and to understand their full potential. Novel fungicides are fundamental to combat plant diseases. Fusarium solani is important plant pathogen. New broad spectrum foliar fungicides against complex II were designed in this study. Complex II, namely succinate dehydrogenase (SDH), or succinate quinone oxidoreductase (SQR) is a multi-subunit enzyme at the crossroads of TCA and ETC at the inner mitochondrial membrane. The need for new and innovative fungicides is driven by resistance management, regulatory hurdles and increasing customer expectations amongst others. Fungicidal activity was assessed for the effect on mycelial growth and spore germination of the fungi using fungicide amended media assay. In mycelial growth assay compounds C10 and C6 were highly active against all the isolates. The compounds C1 and C10 were found most potent in spore germination test. It fully proved that the SDHIs designed in this paper displayed as good inhibitory effects against Fusarium solani.

Keywords: Wilt, Fusarium, SDH, antifungal

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Authors: Rumaisa Shahid, Saad Aziz Durrani, Shameel Pervez, Ibatsam Khokhar

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Food waste is a prevalent issue in Pakistan, with over 40 percent of food discarded annually. Despite their decay, rotting fruits retain residual nutritional value consumed by microorganisms, notably fungi and bacteria. Fungi, preferred for their extracellular enzyme release, are gaining prominence, particularly for pectinase production. This enzyme offers several advantages, including clarifying juices by breaking down pectic compounds. In this study, three Aspergillus flavus isolates derived from decomposed fruits and manure were selected for pectinase production. The primary aim was to isolate fungi from diverse waste sources, identify the isolates and assess their capacity for pectinase production. The identification was done through morphological characteristics with the help of Light microscopy and Scanning Electron Microscopy (SEM). Pectinolytic potential was screened using pectin minimal salt agar (PMSA) medium, comparing clear zone diameters among isolates. Identification relied on morphological characteristics. Optimizing substrate (lemon and orange peel powder) concentrations, pH, temperature, and incubation period aimed to enhance pectinase yield. Spectrophotometry enabled quantitative analysis. The temperature was set at room temperature (28 ºC). The optimal conditions for Aspergillus flavus strain AF1(isolated from mango) included a pH of 5, an incubation period of 120 hours, and substrate concentrations of 3.3% for orange peels and 6.6% for lemon peels. For AF2 and AF3 (both isolated from soil), the ideal pH and incubation period were the same as AF1 i.e. pH 5 and 120 hours. However, their optimized substrate concentrations varied, with AF2 showing maximum activity at 3.3% for orange peels and 6.6% for lemon peels, while AF3 exhibited its peak activity at 6.6% for orange peels and 8.3% for lemon peels. Among the isolates, AF1 demonstrated superior performance under these conditions, comparatively.

Keywords: pectinase, lemon peel, orange peel, aspergillus flavus

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Authors: Ali M. Zaid, Muftah Mayouf, Yahya Said Farouj

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The effect of post emergence herbicides on soil micro-flora and nitrogen fixing bacteria was studied in pea field. Pea (Pisum sativum) was grown and treated with one or a mixture of two of several herbicides 2 weeks after sowing. Soil samples were collected 2 weeks after herbicides application. Average number of colony forming units per gram of soil of bacteria, actinomycetes and fungi were determined. Average number of nodules per plant was obtained at the end of the growing season. The results of the study showed MCPB, Bentazon, MCPB+Fluozifop-p-butyl, Bentazon+Fluozifop-p-butyl, Metribuzin, Flouzifop-p-butyl+Metribuzin, Cycloxydin, and Sethoxydin increased the population of soil fungi, with 4 to 10 times compared with the control. The herbicides used showed no significant effects on nitrogen fixing bacteria. The effects of herbicides on soil bacteria and actinomycetes were different. The study showed the use of herbicides could influence the biological balance of soil microflora, which has an important role in soil fertility and microbial ecosystem.

Keywords: herbicides, post emergence, nitrogen fixing bacteria, environmental systems

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Authors: Eszter J. Tóth, Alexandra Hoffmann, Csaba Vágvölgyi, Tamás Papp

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Members of the genera Curvularia and Bipolaris are closely related melanin producing filamentous fungi; both of them have the teleomorph states in genus Cochliobolus. While Bipolaris species infect only plants and may cause serious agriculture damages, some Curvularia species was recovered from opportunistic human infections. The human pathogenic species typically cause phaeohyphomycoses, i.e. mould infections caused by melanised fungi, which can manifest as invasive mycoses with frequent involvement of the central nervous system in immunocompromised patients or as local infections (e.g. keratitis, sinusitis, and cutaneous lesions) in immunocompetent people. Although their plant-fungal interactions have been intensively studied, there is only little information available about the human pathogenic feature of these fungi. The aim of this study was to investigate the neutrophil granulocytes’ response to hyphal forms of Curvularia and Bipolaris in comparison with the response to Aspergillus. In the present study Curvularia lunata SZMC 23759 and Aspergillus fumigatus SZMC 23245 both isolated from human eye infection, and Bipolaris zeicola BRIP 19582b isolated from plant leaf were examined. Neutrophils were isolated from heparinised venous blood of healthy donors with dextran sedimentation followed by centrifugation over Ficoll and hypotonic lysis of erythrocytes. Viability and purity of the cells were checked with trypan blue and Wright staining, respectively. Infection of neutrophils was carried out with germinated conidia in a ratio of 5:1. Production of hydrogen peroxide, superoxide anion, and nitrogen monoxide was measured both intracellularly and extracellularly in response to the germinated spores with or without the supernatant and after serum treatment. ROS and NOS production of neutrophils in interaction with the three fungi were compared. It is already known that Aspergillus species induce ROS production of neutrophils only after serum treatment. Although, in case of C. lunata, serum opsonisation also induced an intensive production of reactive species, lower level of production was measured in the lack of serum as well. After interaction with the plant pathogenic B. zeicola, amount of reactive species found to be similar with and without serum treatment. The presence of germination supernatant decreased the reactive species production in case of each fungus. Interaction with Curvularia, Bipolaris and Aspergillus species induced different response of neutrophils. It seems that recognition of C. lunata and B. zeicola is independent of serum opsonisation, albeit it increases the level of the produced reactive species in response for C. lunata. The study was supported by the grant LP2016-8/2016.

Keywords: Curvularia, neutrophils, NOS, ROS, serum opsonisation

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Authors: Zaida Perez-Bassart, Berta Polanco-Estibalez, Maria Jose Fabra, Amparo Lopez-Rubio, Antonio Martinez-Abad

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Mushroom production has enormously increased in recent years, not only as food products but also for applications in pharmaceuticals, nutraceuticals, and cosmetics. Consequently, interest in its chemical composition, nutritional value, and therapeutic properties has also increased. Fungi are rich in bioactive compounds such as polysaccharides, polyphenols, glycopeptides, and ergosterol, of great medicinal value, but within polysaccharides, β-glucans are the most prominent molecules. They are formed by D-glucose monomers, linked by β-glucosidic bonds β-(1,3) with side chains linked by β-(1,6) bonds. The number and position of the β-(1,6) branches strongly influence the arrangement of the tertiary structure, which, together with the molecular weight, determine the different attributed bioactivities (immunostimulating, anticancer, antimicrobial, prebiotic, etc.) and physico-chemical properties (solubility, bioaccessibility, viscosity or emulsifying). On the other hand, there is a growing interest in the study of fungi as an alternative source of chitin obtained from the by-products of the fungal industry. In this work, a cascade extraction process using aqueous neutral and alkaline treatments was carried out for Grifola frondosa and Lentinula edodes, and the compositional analysis and functional properties of each fraction were characterized. Interestingly, the first fraction obtained by using aqueous treatment at room temperature was the richest in polysaccharides, proteins, and polyphenols, thus obtaining a greater antioxidant capacity than in the other fractions. In contrast, the fractions obtained by alkaline treatments showed a higher degree of β-glucans purification compared to aqueous extractions but a lower extraction yield. Results revealed the different structural recalcitrance of β-glucans, preferentially linked to proteins or chitin depending on the fungus type, which had a direct impact on the functionalities and bioactivities of each fraction.

Keywords: fungi, mushroom, β-glucans, chitin

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Authors: Birhane Atnafu, Chemeda Abedeta Garbaba, Fikre Lemessa, Abdi Mohammed, Alemayehu Chala

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Essential oil is a bio-pesticide plant product used as an alternative to pesticides in managing plant pests, including fungal pathogens. Thus, the current study aims to investigate the chemical composition and antifungal activities of essential oils (EO) extracted from three aromatic plants i.e., Thymus vulgaris, Coriandrum sativum, and Cymbopogon martini. The leaf parts of those selected plants were collected from the Jimma area and their essential oil was extracted by hydro-distillation method in a Clevenger apparatus. The chemical composition of selected plant essential oil was analyzed by using Gas chromatography-mass spectrometry (GC/MS) and their inhibitory effects were tested in vitro on toxigenic fungi isolated from maize kernel. Chemical analysis results revealed the presence of 32 compounds in C. sativum with Hexanedioic acid, bis (2-ethylhexyl) ester (46. 9%), 2-Decenal, (E)- (12.6), and linalool (8.3%) being the dominant ones. T. vulgaris essential oils constituted 25 compounds, of which thymol (34.4%), o-cymene (17.5%), and Gamma-Terpinene (16.8%) were the major components. Twenty-five compounds were detected in C. martinii of which geraniol (51.4%), Geranyl acetate (14.5%), and Trans – ß-Ocimene (11.7%) were dominant. The EOs of the tested plants had very high antifungal activity (up to 100% efficacy) against Aspergillus flavus, Aspergillus niger, Fusarium graminearum and Fusarium verticillioides in vitro and on maize grains. The antifungal activities of these essential oils were dependent on the major components such as thymol, hexanedioic acid, bis (2-ethylhexyl) ester, and geraniol. The study affirmed the potential of these essential oils controlling as bio-fungicides to manage the effects of potentially toxigenic fungi associated with maize under post-harvest stages. This can reduce the consequences of the health impacts of the mold and toxigenic compounds produced in maize.

Keywords: bio-activity, bio-pesticides, maize, mycotoxin

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Authors: Aboul-Nasr Amal, Sabry Soraya, Sabra Mayada

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The effects of phosphorus amendments and arbuscular mycorrhizal (AM) fungi Glomus intraradices on the sweet basil (Ocimum basilicum L.), chemical composition and percent of volatile oil, and metal accumulation in plants and its availability in soil were investigated in field experiment at two seasons 2012 and 2013 under contaminated soil with Pb and Cu. The content of essential oil and shoot and root dry weights of sweet basil was increased by the application of mineral phosphorus as compared to control. Inoculation with AM fungi reduced the metal concentration in shoot, recording a lowest value of (33.24, 18.60 mg/kg) compared to the control (46.49, 23.46 mg/kg) for Pb and Cu, respectively. Availability of Pb and Cu in soil were decreased after cultivation in all treatments compared to control. However, metal root concentration increased with the inoculation, with highest values of (30.15, 39.25 mg/kg)compared to control (22.01, 33.57mg/kg) for Pb and Cu, respectively. The content of linalool and methyl chavicol in basil oil was significantly increased in all treatments compared to control. We can thus conclude that the AM-sweet basil symbiosis could be employed as an approach to bioremediate polluted soils and enhance the yield and maintain the quality of volatile oil of sweet basil plants.

Keywords: arbuscular mycorrhizal fungus, heavy metals, sweet basil, oil composition

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Authors: Jacqueline T. Madaure, Phatu W. Mashela

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Interactive effects of cucurbitacin-containing phytonematicides and biocontrol agents on growth and nematode suppression on tomato (Solanum lycopersicum) had not been documented. The objective of this study was to determine the interactive effects of Nemafric-BL phytonematicide, Trichoderma harzianum and Steinernema feltiae on growth of tomato plants and suppression of root-knot (Meloidogyne species) nematodes. A 2x2x2 trial was conducted using tomato cv. ‘HTX’ on a field infested with Meloidogyne species. The treatments were applied at commercial rates. At 56 days after treatments, interactions were significant (P ≤ 0.05) for selected plant variables, without significant interactions on nematode variables. In conclusion, results of the current study did not support the combination of the test products for nematode suppression, except that some combinations improved plant growth.

Keywords: cucumis africanus, cucurbitacin b, ethnobotanicals, entomopathogenic nematodes, natural enemies, plant extracts

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Authors: Mohammad A. M. Kewisha

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Xerophilic fungi , which are responsible for many cases of biodeterioration monuments, have been known as an interesting source of antimicrobial compounds. Sixty nine fungal strains, isolated from different localities and species inside Egyptian museums, were screened for antimicrobial activity against some bacterial species and unicellular fungi. The most potent antimicrobial activity was obtained by Asperigillus fumigatus which was identified by ITS4 ……. and showed activity against Staphylococcus aureus with 20 mm and C. albicans with18 mm of inhibition zone. Different parameters were optimized to enhance this activity. The culture grown under stationary conditions for 8 days at 30°C and pH 8 gave the best antimicrobial activity. Moreover, both starch and yeast extract showed the most suitable carbon and nitrogen sources, respectively. The antimicrobial compound was purified and subjected to spectroscopic characterization, which revealed that the antimicrobial compound might be 5,7 ethoxy, 4\,5\ methoxy isorhamnetin -3- O- galactoside. This study suggests that Aspergillus fumagates as a potential candidate offering a better scope for the production, purification and isolation of broad-spectrum antimicrobial compounds. These findings will facilitate the scale-up and further purification to ascertain the compounds responsible for antimicrobial activity, which can be exploited for the treatment of biodeterioration monuments and pharmaceutical applications.

Keywords: antimicrobial activity, asperigillus fumigatus, Identification by ITS4, Staphylococcus aureus, C.albicans

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