Search results for: entomopathogenic fungi

Authors: Amandeep Kaur, Ns Atri, Munruchi Kaur

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Mushrooms have shaped our environment in ways that we are only beginning to understand. The weather patterns, topography, flora and fauna of Punjab state in India create favorable growing conditions for thousands of species of mushrooms, but the complete region was unexplored when it comes to coprophilous mushrooms growing on herbivorous dung. Coprophilous mushrooms are the most specialized fungi ecologically, which germinate and grow directly on different types of animal dung or on manured soil. In the present work, the diversity of coprophilous mushrooms' of Family Psathyrellaceae of the order Agaricales is explored, their relationship to the human world is sketched out, and their supreme significance to life on this planet is revealed. During the investigation, different dung localities from 16 districts of Punjab state have been explored for the collection of material. The macroscopic features of the collected mushrooms were documented on the Field key. The hand cut sections of the various parts of carpophore, such as pileus, gills, stipe and the basidiospores details, were studied microscopically under different magnification. Various authentic publications were consulted for the identification of the investigated taxa. The classification, authentic names and synonyms of the investigated taxa are as per the latest version of Dictionary of Fungi and the MycoBank. The present work deals with the taxonomy of 81 collections belonging to 39 species spread over 05 coprophilous genera, namely Psathyrella, Panaeolus, Parasola, Coprinopsis, and Coprinellus of family Psathyrellaceae. In the text, the investigated taxa have been arranged as they appear in the key to the genera and species investigated. In this work, have been thoroughly examined for their macroscopic, microscopic, ecological, and chemical reaction details. The authors dig deeper to give indication of their ecology and the dung type where they can be obtained. Each taxon is accompanied by a detailed listing of its prominent features and an illustration with habitat photographs and line drawings of morphological and anatomical features. Taxa are organized as per their status in the keys, which allow easy recognition. All the taxa are compared with similar taxa. The study has shown that dung is an important substrate which serves as a favorable niche for the growth of a variety of mushrooms. This paper shows an insight what short-lived coprophilous mushrooms can teach us about sustaining life on earth!

Keywords: abundance, basidiomycota, biodiversity, seasonal availability, systematics

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Authors: Vladimíra Kňazovická, Mária Dovičičová, Miroslava Kačániová, Margita Čanigová

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The aim of the study was to test the storage of bee pollen at room temperature and in cold store, and to describe microorganisms originated from it. Fresh bee pollen originating in West Slovakia was collected in May 2010. It was tested for presence of particular microbial groups using dilution plating method, and divided into two parts with different storage (in cold store and at room temperature). Microbial analyses of pollen were repeated after one year of storage. Several bacterial strains were isolated and tested using Gram staining, for catalase and fructose-6-phosphate-phosphoketolase presence, and by rapid ID 32A (BioMérieux, France). Micromycetes were identified at genus level. Fresh pollen contained coliform bacteria, which were not detected after one year of storage in both ways. Total plate count (TPC) of aerobes and anaerobes and of yeasts in fresh bee pollen exceeded 5.00 log CFU/g. TPC of aerobes and anaerobes decreased below 2.00 log CFU/g after one year of storage in both ways. Count of yeasts decreased to 2.32 log CFU/g (at room temperature) and to 3.66 log CFU/g (in cold store). Microscopic filamentous fungi decreased from 3.41 log CFU/g (fresh bee pollen) to 1.13 log CFU/g (at room temperature) and to 1.89 log CFU/g (in cold store). In fresh bee pollen, 12 genera of micromycetes were identified in the following order according to their relative density: Penicillium > Mucor > Absidia > Cladosporium, Fusarium > Alternaria > Eurotium > Aspergillus, Rhizopus > Emericella > Arthrinium and Mycelium sterilium. After one year at room temperature, only three genera were detected in bee pollen (Penicillium > Aspergillus, Mucor) and after one year in cold store, seven genera were detected (Mucor > Penicillium, Emericella > Aspergillus, Absidia > Arthrinium, Eurotium). From the plates designated for anaerobes, eight colonies originating in fresh bee pollen were isolated. Among them, a single yeast isolate occurred. Other isolates were G+ bacteria, with a total of five rod shaped. In three out of these five, catalase was absent and fructose-6-phosphate-phosphoketolase was present. Bacterial isolates originating in fresh pollen belonged probably to genus Bifidobacterium or relative genera, but their identity was not confirmed unequivocally. In general, cold conditions are suitable for maintaining the natural properties of foodstuffs for a longer time. Slight decrease of microscopic fungal number and diversity was recorded in cold temperatures compared with storage at room temperature.

Keywords: bacteria, bee product, microscopic fungi, biosystems engineering

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Authors: Nuha Mansour Alhazmi, Magda Mohamed Aly, Fardus M. Bokhari, Ahmed Bahieldin, Sherif Edris

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Out of 30 fungal isolates, 2 new isolates were proven to degrade poly-β-hydroxybutyrate (PHB). Enzyme assay for these isolates indicated the optimal environmental conditions required for depolymerase enzyme to induce the highest level of biopolymer degradation. The two isolates were basically characterized at the morphological level as Trichoderma asperellum (isolate S1), and Aspergillus fumigates (isolate S2) using standard approaches. The aim of the present study was to characterize these two isolates at the molecular level based on the highly diverged rRNA gene(s). Within this gene, two domains of the ribosome large subunit (LSU) namely internal transcribed spacer (ITS) and 26S were utilized in the analysis. The first domain comprises the ITS1/5.8S/ITS2 regions ( > 500 bp), while the second domain comprises the D1/D2/D3 regions ( > 1200 bp). Sanger sequencing was conducted at Macrogen (Inc.) for the two isolates using primers ITS1/ITS4 for the first domain, while primers LROR/LR7 for the second domain. Sizes of the first domain ranged between 594-602 bp for S1 isolate and 581-594 bp for S2 isolate, while those of the second domain ranged between 1228-1238 bp for S1 isolate and 1156-1291 for S2 isolate. BLAST analysis indicated 99% identities of the first domain of S1 isolate with T. asperellum isolates XP22 (ID: KX664456.1), CTCCSJ-G-HB40564 (ID: KY750349.1), CTCCSJ-F-ZY40590 (ID: KY750362.1) and TV (ID: KU341015.1). BLAST of the first domain of S2 isolate indicated 100% identities with A. fumigatus isolate YNCA0338 (ID: KP068684.1) and strain MEF-Cr-6 (ID: KU597198.1), while 99% identities with A. fumigatus isolate CCA101 (ID: KT877346.1) and strain CD1621 (ID: JX092088.1). Large numbers of other T. asperellum and A. fumigatus isolates and strains showed high level of identities with S1 and S2 isolates, respectively, based on the diversity of the first domain. BLAST of the second domain of S1 isolate indicated 99 and 100% identities with only two strains of T. asperellum namely TR 3 (ID: HM466685.1) and G (ID: KF723005.1), respectively. However, other T. species (ex., atroviride, hamatum, deliquescens, harzianum, etc.) also showed high level of identities. BLAST of the second domain of S2 isolate indicated 100% identities with A. fumigatus isolate YNCA0338 (ID: KP068684.1) and strain MEF-Cr-6 (ID: KU597198.1), while 99% identities with A. fumigatus isolate CCA101 (ID: KT877346.1) and strain CD1621 (ID: JX092088.1). Large numbers of other A. fumigatus isolates and strains showed high level of identities with S2 isolate. Overall, the results of molecular characterization based on rRNA diversity for the two isolates of T. asperellum and A. fumigatus matched those obtained by morphological characterization. In addition, ITS domain proved to be more sensitive than 26S domain in diversity profiling of fungi at the species level.

Keywords: Aspergillus fumigates, Trichoderma asperellum, PHB, degradation, BLAST, ITS, 26S, rRNA

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Authors: M. Lamamra, H. Laouer, A. Adjaoud, Sahli Farida

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Essential oils can have significant antimicrobial activities and can successfully replace antibiotics that show their ineffectiveness against resistant germs. The chemical composition of the essential oil obtained by hydrodistillation from the aerial part of Daucus aristidis (Apiaceae) at the pre-flowering stage was investigated for the first time, by GC and GC-MS and evaluated for in vitro antimicrobial activity by the disk diffusion method. The Main components of D. aristidis oil were α-pinene (20.13%), cedrol (20.11%), and E- asarone (18.53%). The oil exhibited an antibacterial activity against almost strains tested except for Klebsiella pneumoniae ATCC 700603 K6 and Enterococcus faecalis ATCC 49452, the oil of D. aristidis had no activity against all fungi tested.

Keywords: α-pinene, antimicrobial activity, Daucus aridtidis, essential oil

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Authors: Leonardo Parra, Daniel Martínez, Jorge Pizarro, Fernando Ortega, Manuel Chacón-Fuentes, Andrés Quiroz

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Control of Dalaca pallens or blackworms, one of the most important hypogeous pest in grassland in southern Chile, is based on the use of broad-spectrum insecticides such as organophosphates and pyrethroids. However, the rapid development of insecticide resistance in field populations of this insect and public concern over the environmental impact of these insecticides has resulted in the search for other control methods. Specifically, the use of endophyte fungi for controlling pest has emerged as an interesting and promising strategy. Endophytes from ryegrass (Lolium perenne), establish a biotrophic relationship with the host, defined as mutualistic symbiosis. The plant-fungi association produces alkaloids where peramine is the main toxic substance against Listronotus bonariensis, the most important epigean pest of ryegrass. Nevertheless, the effect of peramina on others pest insects, such as D. pallens, to our knowledge has not been studied, and also its possible metabolization in the body of the larvae. Therefore, we addressed the following research question: Do larvae of D. pallens store peramine after consumption of ryegrass endophyte infected (E+)? For this, specimens of blackworms were fed with ryegrass plant of seven experimental lines and one commercial cultivar endophyte free (E-) sown at the Instituto de Investigaciones Agropecuarias Carillanca (Vilcún, Chile). Once the feeding period was over, ten larvae of each treatment were examined. Individuals were dissected, and their gut was removed to exclude any influence of remaining material. The rest of the larva's body was dried at 60°C by 24-48 h and ground into a fine powder using a mortar. 25 mg of dry powder was transferred to a microcentrifuge tube and extracted in 1 mL of a mixture of methanol:water:formic acid. Then, the samples were centrifuged at 16,000 rpm for 3 min, and the supernatant was colected and injected in the liquid chromatography of high resolution (HPLC). The results confirmed the presence of peramine in the larva's body of D. pallens. The insects that fed the experimental lines LQE-2 and LQE-6 were those where peramine was present in high proportion (0.205 and 0.199 ppm, respectively); while LQE-7 and LQE-3 obtained the lowest concentrations of the alkaloid (0.047 and 0.053 ppm, respectively). Peramine was not detected in the insects when the control cultivar Jumbo (E-) was tested. These results evidenced the storage and metabolism of peramine during consumption of the larvae. However, the effect of this alkaloid present in 'future ryegrass cultivars' (LQE-2 and LQE-6) on the performance and survival of blackworms must be studied and confirmed experimentally.

Keywords: blackworms, HPLC, alkaloid, pest

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Authors: Falana Yetunde Olaitan, Ijah U. J. J, Solebo Shakirat O.

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Genetically modified crops are already phenomenally successful and are grown worldwide in more than eighteen countries on more than 67 million hectares. Nigeria, in October 2018, approved Bacillus thuringiensis (Bt) cotton and maize; therefore, the need to carry out environmental risk assessment studies. A total of 15 4L octagonal ceramic pots were filled with 4kg of soil and placed on the bench in 2 rows of 10 pots each and the 3rd row of 5 pots, 1st-row pots were used to plant GM cotton seeds, while the 2nd-row pots were used for non-GM cotton seeds and the 3rd row of 5 pots served as control, all in the screen house. Soil samples for metagenomic DNA extraction were collected at random and at the monthly interval after planting at a distance of 2mm from the plant’s root and at a depth of 10cm using a sterile spatula. Soil samples for physicochemical analysis were collected before planting and after harvesting the GM and non-GM crops as well as from the control soil. The DNA was extracted, quantified and sequenced; Sample 1A (DNA from GM cotton Soil at 1st interval) gave the lowest sequence read with 0.853M while sample 2B (DNA from GM cotton Soil at 2nd interval) gave the highest with 5.785M, others gave between 1.8M and 4.7M. The samples treatment were grouped into four, Group 1 (GM cotton soil from 1 to 3 intervals) had between 800,000 and 5,700,000 strains of microbes (SOM), Group 2 (non GM cotton soil from 1 to 3 intervals) had between 1,400,600 and 4,200,000 SOM, Group 3 (control soil) had between 900,000 and 3,600,000 SOM and Group 4 (initial soil) had between 3,700,000 and 4,000,000 SOM. The microbes observed were predominantly bacteria (including archaea), fungi, dark matter alongside protists and phages. The predominant bacterial groups were the Terrabacteria (Bacillus funiculus, Bacillus sp.), the Proteobacteria (Microvirga massiliensis, sphingomonas sp.) and the Archaea (Nitrososphaera sp.), while the fungi were Aspergillus fischeri and Fusarium falciforme. The comparative analysis between groups was done using JACCARD PERMANOVA beta diversity analysis at P-value not more than 0.76 and there was no significant pair found. The pH for initial, GM cotton, non-GM cotton and control soil were 6.28, 6.26, 7.25, 8.26 and the percentage moisture was 0.63, 0.78, 0.89 and 0.82, respectively, while the percentage Nitrogen was observed to be 17.79, 1.14, 1.10 and 0.56 respectively. Other parameters include, varying concentrations of Potassium (0.46, 1,284.47, 1,785.48, 1,252.83 mg/kg) and Phosphorus (18.76, 17.76, 16.87, 15.23 mg/kg) were recorded for the four treatments respectively. The soil consisted mainly of silt (32.09 to 34.66%) and clay (58.89 to 60.23%), reflecting the soil texture as silty – clay. The results were then tested with ANOVA at less than 0.05 P-value and no pair was found to be significant as well. The results suggest that the GM crops have no significant effect on microbial ecology and physicochemical properties of the soil and, in turn, no direct or indirect effects on human health.

Keywords: genetically modified crop, microbial ecology, physicochemical properties, metagenomics, DNA, soil

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Authors: Jovie Esquivias Nicolas, Ron Aldrin Lontoc Austria, Crisabelle Belleza Bautista, Mariane Chiho Espinosa Bundalian, Owwen Kervy Del Rosario Castillo, Mary Angelyn Mercado Dela Cruz, Heinrich Theraja Recana De Luna, Chriscell Gipanao Eustaquio, Desiree Laine Lauz Gilbas, Jordan Ignacio Legaspi, Larah Denise David Madrid, Charles Linelle Malapote Mendoza, Hazel Maxine Manalad Reyes, Carl Justine Nabora Saberdo, Claire Mae Rendon Santos

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In response to the global race in discovering the next advanced sustainable material that will reduce our ecological footprint, the researchers aimed to create a masonry unit which is competent in physical edifices and other constructional facets. From different proven researches, mycelium has been concluded that when dried can be used as a robust and waterproof building material that can be grown into explicit forms, thus reducing the processing requirements. Hypothesizing inclusive measures to attest fungi’s impressive structural qualities and absorbency, the researchers projected to perform comparative analyses in creating mycelium bricks from mushroom spores of G. lucidum. Three treatments were intended to classify the most ideal concentration of clay and substrate fixings. The substrate bags fixed with 30% clay and 70% mixings indicated highest numerical frequencies in terms of full occupation of fungal mycelia. Subsequently, sorted parts of white portions from the treatment were settled in a thermoplastic mold and burnt. Three proportional concentrations of cultivated substrate and cement were also prioritized to gather results of variation focused on the weights of the bricks in the Water Absorption Test and Durability Test. Fungal inoculums with solutions of cement showed small to moderate amounts of decrease and increase in load. This proves that the treatments did not show any significant difference when it comes to strength, efficiency and absorption capacity. Each of the concentration is equally valid and could be used in supporting the worldwide demands of creating numerous bricks while also taking into consideration the recovery of our nature.

Keywords: mycelium, fungi, fungal mycelia, durability test, water absorption test

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Authors: Judit Perjessy, Zsolt Zalan, Ferenc Hegyi, Eniko Horvath-Szanics, Krisztina Takacs, Andras Nagy, Adel Klupacs, Erika Koppany-Szabo, Zhirong Wang, Kaituo Wang, Muying Du, Jianquan Kan

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The post-harvest diseases cause great economic losses in the fruit and vegetables; the prevention of these deterioration has great importance. Against the fungi, which cause most of the diseases, are extensively used the fungicides. However, there are increasing consumer concerns over the presence of pesticide residues in food. An alternative and in recent years, increasingly studied method for the prevention of the diseases is biocontrol, where antagonistic microorganisms are used for the control of fungi. The genera of Lactobacillus is well known and extensively studied, but its applicability as biocontrol agents in post-harvest preservation of fruit and vegetables is poorly investigated. However these bacteria can be found on the surface of the plants and have great antimicrobial activity. In our study we have investigated the chitinase activity, the antifungal effect and the applicability of several Lactobacillus strains to select potential biocontrol agents. We investigated the determination of the environmental parameters of a gene (encoding chitinase) expression and we also investigated the relationship between actual antifungal activity and potential chitinase activity. Mixed cultures were also developed to enhance the antifungal activity and determined the optimal mold spore and bacteria concentration ratio for the appropriate efficacy. Five Lactobacillus strains (L. acidophilus N2, L. delbrueckii subsp. bulgaricus B397, L. sp. 2231, L. sake subsp. sake 2471, L. buchneri 1145) possess chitinase-coding gene from the 43 investigated Lactobacillus strains. Proteins with similar molecular weight and separation properties like bacterial chitinases were detected from these strains, which also possess chitin-binding property. Nevertheless, they were inactive, lacks the chitinolytic activity. In point of the cumulative activity of inhibition, our results showed that certain strains were statistically significant in a positive direction compared to other strains, e.g., L. rhamnosus VT1 and L. Casey 154 have shown great general antifungal effect against 11 molds from the genera Penicillium and Botrytis and isolated from spoiled fruit and vegetables. Also, some mixed cultures (L. rhamnosus VT1 - L. Plantarum 299v) showed significant antifungal effects against the indigenous molds on the surface of apple fruit during the industrial storage experiment. Thus, they could be promising for post-harvest biopreservation.

Keywords: biocontrol, chitinase, Lactobacillus, post-harvest

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Authors: In-Lee Choi, Sung Mi Hong, Min Jae Jeong, Jun Pill Baek, Ho-Min Kang

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Gomchwi (Ligularia fischeri) is grown in the wetland of the deep mountains in Korea and East Asia and has properties that are, inflammation control, whitening, antimutagenic and antigenotoxic. Jangajji is a type of pickle in Korean fermented food which is made by pickling or marinating vegetables in a sauce, such as soy sauce, chili pepper paste, soybean paste, or diluted vinegar for a long period of time. Handaeri-gomchwi jangajii is generally packed a film that has very low or no gas permeability in the Korean domestic market, so packages have a risk of swelling or bursting as a result of internal gas generation during storage or sale This study was conducted to improve secure distribution of Handaeri-gomchwi (Ligularia fischeri var. spiciformis Nakai) Jangajji using laser ablation OTR (oxygen transmission rate) films. Handaeri-gomchwi cultivated in Yangu, Gangwon province, Republic of Korea (Ligularia fischeri var. spiciformis Nakai) was processed in to Jangajji using soy sauce. They were packed by different OTR films, and were stored for 90 days in 7℃(10,000 cc, 20,000 cc, 40,000 cc and 80,000 cc O2/m²• day • atm), 20 days in 20℃ (10,000 cc, 30,000 cc, 70,000 cc and 100,000 cc) and compared with the control film(PP film, 1,300cc). The fresh weight loss, carbon dioxide, oxygen, and ethylene concentrations of Handaeri–gomchwi packages were measured during storage. On the final day of storage, incidence rate of fungi, pH, salinity, firmness, and off-flavor were measured. The fresh weight loss rate of Handaeri–gomchwi was less than 2.0% in 10,000cc OTR films at two different storage periods and temperatures. At 80,000cc(7℃) and 100,000cc(20℃), carbon dioxide contents were 2.0% and 6.4% respectively, whereas the control treatment had the highest concentration. Which was 35%(20℃) and 15%(7℃) , that resulted the packages to swell during storage. The control treatment Showed the lowest oxygen concentration at 2.5% in 7℃ and 0.8% in 20℃. Packages in 7℃ (0.3-1.7μL/L) showed very lower ethylene concentration than in 20℃(10-25μL/L), they also had no significant relation. On the final storage day, fungi were found in every film at both temperatures, except the 10,000cc, as oxygen permeability increased so did the pH, while the salinity decreased. Firmness and off-flavor Showed the best results at 10,000cc in both temperatures best result at 10,000cc in both temperature. Following the results, 10,000cc film is the most reasonable treat in storing Handaeri–gomchwi. For it had a suitable oxygen transmission rate, which prevents billowing, and maintained good qualities in both temperatures.

Keywords: carbon dioxide, Korean pickle, marketable, oxygen

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Authors: Raphael C. Zero, Marita V. Cardozo, Thiago A. S. S. Rocha, Mariana T. Kihara, Fernando A. Ávila, Fabrício S. Oliveira

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There are several fixative and preservative solutions for use on cadavers, many of them using formaldehyde as the fixative or anatomical part preservative. In some countries, such as Brazil, this toxic agent has been increasingly restricted. The objective of this study was to microbiologically identify and quantify the key agents in tanks containing 96GL ethanol or sodium chloride solutions, used respectively as fixatives and preservatives of cat cadavers. Eight adult cat corpses, three females and five males, with an average weight of 4.3 kg, were used. After injection via the external common carotid artery (120 ml/kg, 95% 96GL ethyl alcohol and 5% pure glycerin), the cadavers were fixed in a plastic tank with 96GL ethanol for 60 days. After fixing, they were stored in a 30% sodium chloride aqueous solution for 120 days in a similar tank. Samples were collected at the start of the experiment - before the animals were placed in the ethanol tanks, and monthly thereafter. The bacterial count was performed by Pour Plate Method in BHI agar (Brain Heart Infusion) and the plates were incubated aerobically and anaerobically for 24h at 37ºC. MacConkey agar, SPS agar (Sulfite Polymyxin Sulfadizine) and MYP Agar Base were used to isolate the microorganisms. There was no microbial growth in the samples prior to alcohol fixation. After 30 days of fixation in the alcohol solution, total aerobic and anaerobic (<1.0 x 10 CFU/ml) were found and Pseudomonas sp., Staphylococcus sp., Clostridium sp. were the identified agents. After 60 days in the alcohol fixation solution, total aerobes (<1.0 x 10 CFU/ml) and total anaerobes (<2.2 x 10 CFU/mL) were found, and the identified agents were the same. After 30 days of storage in the aqueous solution of 30% sodium chloride, total aerobic (<5.2 x 10 CFU/ml) and total anaerobes (<3.7 x 10 CFU/mL) were found and the agents identified were Staphylococcus sp., Clostridium sp., and fungi. After 60 days of sodium chloride storage, total aerobic (<3.0 x 10 CFU / ml) and total anaerobes (<7.0 x 10 CFU/mL) were found and the identified agents remained the same: Staphylococcus sp., Clostridium sp., and fungi. The microbiological count was low and visual inspection did not reveal signs of contamination in the tanks. There was no strong odor or purification, which proved the technique to be microbiologically effective in fixing and preserving the cat cadavers for the four-month period in which they are provided to undergraduate students of University of Veterinary Medicine for surgery practice. All experimental procedures were approved by the Municipal Legal Department (protocol 02.2014.000027-1). The project was funded by FAPESP (protocol 2015-08259-9).

Keywords: anatomy, fixation, microbiology, small animal, surgery

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Authors: Brahim Mazian, Anne Bergeret, Jean-Charles Benezet, Sandrine Bayle, Luc Malhautier

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Hemp fibers are increasingly used as reinforcements in polymer matrix composites due to their competitive performance (low density, mechanical properties and biodegradability) compared to conventional fibres such as glass fibers. However, the huge variation of their biochemical, physical and mechanical properties limits the use of these natural fibres in structural applications when high consistency and homogeneity are required. In the hemp industry, traditional processes termed field retting are commonly used to facilitate the extraction and separation of stem fibers. This retting treatment consists to spread out the stems on the ground for a duration ranging from a few days to several weeks. Microorganisms (fungi and bacteria) grow on the stem surface and produce enzymes that degrade pectinolytic substances in the middle lamellae surrounding the fibers. This operation depends on the weather conditions and is currently carried out very empirically in the fields so that a large variability in the hemp fibers quality (mechanical properties, color, morphology, chemical composition…) is resulting. Nonetheless, if controlled, retting might be favorable for good properties of hemp fibers and then of hemp fibers reinforced composites. Therefore, the present study aims to investigate the influence of controlled retting within a designed environmental chamber (lab-scale pilot unit) on the quality of the hemp fibres harvested at the seed maturity growth stage. Various assessments were applied directly on fibers: color observations, morphological (optical microscope), surface (ESEM), biochemical (gravimetry) analysis, spectrocolorimetric measurements (pectins content), thermogravimetric analysis (TGA) and tensile testing. The results reveal that controlled retting leads to a rapid change of color from yellow to dark grey due to development of microbial communities (fungi and bacteria) at the stem surface. An increase of thermal stability of fibres due to the removal of non-cellulosic components along retting is also observed. A separation of bast fibers to elementary fibers occurred with an evolution of chemical composition (degradation of pectins) and a rapid decrease in tensile properties (380MPa to 170MPa after 3 weeks) due to accelerated retting process. The influence of controlled retting on the biocomposite material (PP / hemp fibers) properties is under investigation.

Keywords: controlled retting, hemp fibre, mechanical properties, thermal stability

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Authors: M. N. Alo, U. C. C. Egbule, J. O. Orji, C. J. Aneke

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Microbiological analysis of soil from Onu-Ebonyi Izzi local government area of Ebonyi State, Nigeria contaminated with inorganic fertilizer was carried out with a view to determine the effect of the fertilizer on the microbial flora of the soil. soil samples were analyzed for microbial burden. the result showed that the following organisms were isolated with their frequency of their occurrence as follows:pseudomonas species (33.3%) and aspergillus species (54.4%) had the highest frequncy of occurence in the whole sample of batches, while streptococcus species had 6.0% and Geotrichum species (5.3%) had the least and other predominant microorganism isolated: bacillus species,staphylococcus species and vibrio species, Escherichia species, rhzizopus species, mucor species and fusaruim species. From the result, it could be concluded that the soil was contaminated and this could affect adversely the fertility of the soil .

Keywords: soil, bacteria, fungi, inorganic fertilizer, Onu- Ebonyi

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Authors: P. Chaijak, M. Lertworapreecha, C. Sukkasem

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Extracellular laccases are copper-containing microbial enzymes with many industrial biotechnological applications. This study evaluated the ability of nutrients in coconut coir to enhance the yield of extracellular laccase of Galactomyces reesii IFO 10823 and develop a co-culture between this yeast and other filamentous fungi isolated from the fungus comb of Macrotermes sp. The co-culture between G. reesii IFO 10823 and M. indicus FJ-M-5 (G3) gave the highest activity at 580.20 U/mL. When grown in fermentation media prepared from coconut coir and distilled water at 70% of initial moisture without supplement addition, G3 produced extracellular laccase of 113.99 U/mL.

Keywords: extracellular laccase, production, yeast, natural inducer

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Authors: C. Recchia, M. Bourel, B. Recchia

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Microorganisms (viruses, bacteria, fungi) are responsible for most communicable diseases, threatening human health. For domestic use, chemical agents are often criticized because of their potential dangerousness, and natural solutions are needed. Application of the “dry microfine steam” (DMS) technology was tested on a selection of common pathogens (SARS-CoV-2, enterovirus EV-71, human coronavirus 229E, E. coli, S. aureus, C. albicans), on different innate surfaces, for 5 to 10 seconds. Quantification of the remaining pathogens was performed, and the reduction rates ranged from 99.8% (S. aureus on plastic) to over 99.999%. DMS showed high efficacy in the elimination of common microorganisms and could be seen as a natural alternative to chemical agents to improve domestic hygiene.

Keywords: steam, SARS-CoV-2, bactericidal, virucidal, fungicidal, sterilization

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Authors: Sulamain Alharbi, Mohammad Khiyami, Reda Amasha, Bassam Al-Johny, Hesham Khalil, Milton Wainwrigh

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Nematodes, Rotifers and Tardigrades (NRT) are extreme-tolerant invertebrates which can survive long periods of stasis brought about by extreme drying and cold. They can also resist the effects of UV radiation, and as a result could act as vehicles for the panspermic transfer of microorganisms. Here we show that NRT contain a variety of bacteria and fungi within their bodies in which environment they could be protected from the extremes of the space and released into new cosmic environments. Diatoms were also shown to contain viable alga and Escherichia coli and so could also act as panspermic vehicles for the transfer of these and perhaps other microbes through space. Although not studied here, NRT, and possibly diatoms, also carry protozoa and viruses within their bodies and could act as vehicles for the panspermic transfer of an even wider range of microbes than shown here.

Keywords: extromophiles, diatoms, panspermia, survival in space

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Authors: Abdol-Hassan Doulah

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In this study, antimicrobial activity of essential oil and ethyl acetate and ether extracts of S. nemorosa were examined against some species of bacteria and fungi. The essential oil of the aerial part of S. nemorosa was examined by GC and GC-MS. In the essential oil of S. nemorosa 26 Compounds have been identified. 2-Nonanone (44.09 %), 2-Undecanone (33.79 %), E-Caryophyllene (3.74 %) and 2-Decanone (2.89 %) were the main components of the essential oil. The essential oil analysis showed greatest antimicrobial activity against Staphylococcus epidermidis (5.3 μg/ml) and S. cerevisiae (9.3 μg/ml). The ethyl acetate showed greatest antimicrobial activity against Bacillus subtilis (106.7 μg/ml), Candida albicans (5.3 μg/ml) and ether extract showed greatest antimicrobial activity against Klebseilla pneumoniae (10.7 μg/ml) and Saccharomyces cerevisiae (10.7 μg/ml). In conclusion, we suggest that the antimicrobial activity of S. nemorosa may be due to its content of germacrene and linalool.

Keywords: antibacterial activity, antifungal activity, Salvia nemorosa L., essential oils, biological activity

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Authors: Sulaiman B. Ali Alharbi, Bassam H. Mashat, Naif Abdullah Al-Harbi, Milton Wainwright, Abeer S. Aloufi, Sulamain Alnaimat

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Bismuth salicylate was found to inhibit the growth of a range of bacteria and yeast, Candida albican. In general the growth of bacteria did not result in the increase in bismuth solubilisation, in contrast, bismuth solubilisation increased following the growth of C. albicans. A significant increase in the biomass (dry weight) of Aspergillus niger and Aspergillus oryzae occurred in vitro when these fungi were grown in the presence of bismuth salicylate. Biomass increase occurred over a range of bismuth compound additions, which in the case of A. oryzae was associated with the increase in the solubilisation of the insoluble bismuth compounds.

Keywords: bacterial inhibition, fungal growth stimulation, medical uses of bismuth, yeast inhibition

Procedia PDF Downloads 342

Authors: F. Pal-Fam, S. Keszthelyi

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Phytosanitary condition of sunflower (Helianthus annuus L.) was endangered by several phytopathogenic agents, mainly microfungi, such as Sclerotinia sclerotiorum, Diaporthe helianthi, Plasmopara halstedtii, Macrophomina phaseolina and so on. There are more agrotechnical and chemical technologies against them, for instance, tolerant hybrids, crop rotations and eventually several in-crop chemical treatments. There are different fungicide treatment methods in sunflower in Hungarian agricultural practice in the quest of obtaining healthy and economic plant products. Besides, there are many choices of useable active ingredients in Hungarian sunflower protection. This study carried out into the examination of the effect of five different fungicide active substances (found on the market) and three different application modes (early; late; and early and late treatments) in a total number of 9 sample plots, 0.1 ha each other. Five successive vegetation periods have been investigated in long term, between 2013 and 2017. The treatments were: 1)untreated control; 2) boscalid and dimoxystrobin late treatment (July); 3) boscalid and dimoxystrobin early treatment (June); 4) picoxystrobin and cyproconazole early treatment; 5) picoxystrobin and cymoxanil and famoxadone early treatment; 6) picoxystrobin and cyproconazole early; cymoxanil and famoxadone late treatments; 7) picoxystrobin and cyproconazole early; picoxystrobin and cymoxanil and famoxadone late treatments; 8) trifloxystrobin and cyproconazole early treatment; and 9) trifloxystrobin and cyproconazole both early and late treatments. Due to the very different yearly weather conditions different phytopathogenic fungi were dominant in the particular years: Diaporthe and Alternaria in 2013; Alternaria and Sclerotinia in 2014 and 2015; Alternaria, Sclerotinia and Diaporthe in 2016; and Alternaria in 2017. As a result of treatments ‘infection frequency’ and ‘infestation rate’ showed a significant decrease compared to the control plot. There were no significant differences between the efficacies of the different fungicide mixes; all were almost the same effective against the phytopathogenic fungi. The most dangerous Sclerotinia infection was practically eliminated in all of the treatments. Among the single treatments, the late treatment realised in July was the less efficient, followed by the early treatments effectuated in June. The most efficient was the double treatments realised in both June and July, resulting 70-80% decrease of the infection frequency, respectively 75-90% decrease of the infestation rate, comparing with the control plot in the particular years. The lowest yield quantity was observed in the control plot, followed by the late single treatment. The yield of the early single treatments was higher, while the double treatments showed the highest yield quantities (18.3-22.5% higher than the control plot in particular years). In total, according to our five years investigation, the most effective application mode is the double in-crop treatment per vegetation time, which is reflected by the yield surplus.

Keywords: fungicides, treatments, phytopathogens, sunflower

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Authors: Nilüfer Yıldız Varan

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The antimicrobial activities of chitosan against various bacteria and fungi are well known, and the antimicrobial activity of chitosan depends on pH. This study investigates the antimicrobial activity at different pH levels. Nylon 6,6 fabrics were treated with different chitosan solutions. Additionally, samples were treated also in basic conditions to see the antimicrobial activities. AATCC Test Method 100 was followed to evaluate the antimicrobial activity using Staphylococcus aureus ATCC 6538 test inoculum. The pH of the chitosan solutions was controlled below 6.5 since chitosan shows its antimicrobial activity only in acidic conditions because of its poor solubility above 6.5. In basic conditions, the samples did not show any antimicrobial activity. It appears from SEM images that the bonded chitosan in the structures exists. In acidic media (ph < 6.5), all samples showed antimicrobial activity. No correlation was found between pH levels and antimicrobial activity in acidic media.

Keywords: chitosan, nylon 6, 6, crosslinking, pH stability, antimicrobial

Procedia PDF Downloads 219

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

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

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

Procedia PDF Downloads 523

Authors: Berk Kılıç, Ömer Aydın, Kerem Mestani, Defne Uzun

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In agriculture and farming, pesticides are frequently used to kill off or fend off any pests (bugs, bacteria, fungi, etc.). However, traditional pesticides have proven to have harmful effects on both the environment and the human body, such as hazards in the endocrine, neurodevelopmental, and reproductive systems. This experiment aims to test the effectiveness of organic/bio-pesticides (environmentally friendly pesticides) compared to traditional pesticides. Black pepper and garlic will be used as biopesticides in this experiment. The results support that organic farming applying organic pesticides operates through non-toxic mechanisms, offering minimal threats to human well-being and the environment. Consequently, consuming organic produce can significantly diminish the dangers associated with pesticide intake. In this study, method is introduced to reduce pesticide-related risks by promoting organic farming techniques within organic/bio-pesticide usage.

Keywords: pesticide, garlic, black pepper, bio-pesticide

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Authors: H. Nacef, L. Larous

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Salin soils occupy about 7% of land area; they are characterized by unsuitable physical conditions for the growth of living organisms. However, researches showed that some microorganisms especially fungi are able to grow and adapt to such extreme conditions; it is due to their ability to develop different physiological mechanisms in their adaptation. The aim of this study is to identify qualitatively the osmolytes that the biotechnological important fungus A. oryzae accumulated and/or produced in its adaptation, which they were detected by Thin-layer chromatography technique (TLC) using several systems, from different media (Wheat brane, MNM medium and MM medium). The results showed that The moderately halotolerant fungus A. oryzae, accumulates mixture of molecules, containing polyols and sugars , some amino acids in addition to some molecules which were not defined. Wheat bran was the best medium for the extraction of these molecules, where the proportion was 85.71%, followed by MNM medium 64.28%, then the minimum medium MM 14.28%. Properties of osmolytes are becoming increasingly useful in molecular biology, agriculture pharmaceutical, medicinal, and biotechnological interests.

Keywords: salinity, aspergillus oryzae, halo tolerance, osmolytes, compatible solutes

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Authors: Ivana Brzonova, Asina Fnu, Alena Kubatova, Evguenii Kozliak, Yun Ji

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Bio-decomposition of lignin by Basidiomycetes in the presence of dimethyl sulfoxide (DMSO) was investigated. The addition of 3-5 vol% DMSO to lignin aqueous media significantly increased the lignin solubility based on UV absorbance. After being dissolved in DMSO, the thermal evolution profile also changed significantly, yielding more high-MW organic carbon at the expense of recalcitrant elemental carbon. Medical fungi C. versicolor, G. lucidum and P. pulmonarius, were observed to grow on the lignin in media containing up to 15 vol. % DMSO. Further detailed product characterization by chromatographic methods corroborated these observations, as more low-MW phenolic products were observed with DMSO as a co-solvent. These results may be explained by the high solubility of lignin in DMSO; thus, the addition of DMSO to the medium increases the lignin availability for microorganisms. Some of these low-MW phenolic products host a big potential to be used in medicine. No significant inhibition of enzymatic activity (laccase, MnP, LiP) was observed by the addition of up to 3 vol% DMSO.

Keywords: basidiomycetes, bio-degradation, dimethyl sulfoxide, lignin

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Authors: Indriyani Nur, Yusnaini

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Lobster cultures have failed because of mortalities associated with parasitic and fungal infections. Monitoring of spawned eggs and larva of bamboo lobsters, Panulirus versicolour, and ornate lobsters, P. ornatus, in a hatchery, was conducted in order to characterize fungal and parasitic diseases of eggs and larva. One species of protozoan parasite (Vorticella sp.) was identified from larvae while two species of fungi (Lagenidium sp. and Haliphthoros sp.) were found on eggs. Furthermore, adult lobsters cultured in floating net cage had burning-like diseases on their pleopod, uropod, and telson. Histopathological samples were collected for parasite and tissue changes. There were two parasites found to infect lobsters on external body and gill which are Octolasmis sp. and Oodinium sp. Histopathology showed tissue changes which are necrosis on hepatopancreas, necrosis in the gills and around the uropods and telson.

Keywords: fungal, histopathology, lobster, parasite, infection

Procedia PDF Downloads 295

Authors: Azhar Alhasawi, Vasu D. Appanna

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Chitin, the second most abundant bio-polymer in nature after cellulose, composed of β (1→4) linked N-acetylglucosamine (GlcNAc), is a major structural component in the cell walls of fungi and the shells of crustaceans. Chitin and its derivatives are gaining importance of economic value due to its biological activity and its industrial and biomedical applications. There are several methods to hydrolyze chitin to NAG, but they are typically expensive and environmentally unfriendly. Chitinase which catalyzes the breakdown of chitin to NAG has received much attention owing to its various applications in biotechnology. The presented research examines the ability of the versatile soil microbe, Pseudomonas fluorescens grown in chitin medium to produce chitinase and a variety of value-added products under abiotic stress. We have found that with high pH, Pseudomonas fluorescens enable to metabolize chitin more than with neutral pH and the overexpression of chitinase was also increased. P-dimethylaminobenzaldehyde (DMAB) assay for NAG production will be monitored and a combination of sodium dodecyl polyacrylamide gels will be used to monitor the proteomic and metabolomic changes as a result of the abiotic stress. The bioreactor of chitinase will also be utilized.

Keywords: Pseudomonas fluorescens, chitin, DMAB, chitinase

Procedia PDF Downloads 355

Authors: V. Ivanov, J. Chu, V. Stabnikov

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Low cost, sustainable, and environmentally friendly microbial cements, grouts, polysaccharides and bioplastics are useful in construction and geotechnical engineering. Construction-related biotechnologies are based on activity of different microorganisms: urease-producing, acidogenic, halophilic, alkaliphilic, denitrifying, iron- and sulphate-reducing bacteria, cyanobacteria, algae, microscopic fungi. The bio-related materials and processes can be used for the bioaggregation, soil biogrouting and bioclogging, biocementation, biodesaturation of water-satured soil, bioencapsulation of soft clay, biocoating, and biorepair of the concrete surface. Altogether with the most popular calcium- and urea based biocementation, there are possible and often are more effective such methods of ground improvement as calcium- and magnesium based biocementation, calcium phosphate strengthening of soil, calcium bicarbonate biocementation, and iron- or polysaccharide based bioclogging. The construction-related microbial biotechnologies have a lot of advantages over conventional construction materials and processes.

Keywords: ground improvement, biocementation, biogrouting, microorganisms

Procedia PDF Downloads 229

Authors: Mohamed E. Khalifa, Adil A. Gobouri

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

Keywords: thiophene, 2-aminothiazole, compounds, antioxidant, antitumor, antimicrobial

Procedia PDF Downloads 344

Authors: Leonardo Parra, Manuel Chacón-Fuentes, Andrés Quiroz

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One of the most important factors in beef and dairy production in the world as well as also in Chile, is related to the correct choice of cultivars or mixtures of forage grasses and legumes to ensure high yields and quality of grassland. However, a great problem is the persistence of the grasses as a result of the action of different hypogeous as epigean pests. The complex insect pests associated with grassland include white grubs (Hylamorpha elegans, Phytoloema herrmanni), blackworm (Dalaca pallens) and Argentine stem weevil (Listronotus bonariensis). In Chile, the principal strategy utilized for controlling this pest is chemical control, through the use of synthetic insecticides, however, underground feeding habits of larval and flight activity of adults makes this uneconomic method. Furthermore, due to problems including environmental degradation, development of resistance and chemical residues, there is a worldwide interest in the use of alternative environmentally friendly pest control methods. In this sense, in recent years there has been an increasing interest in determining the role of endophyte fungi in controlling epigean and hypogeous pest. Endophytes from ryegrass (Lolium perenne), establish a biotrophic relationship with the host, defined as mutualistic symbiosis. The plant-fungi association produces a “cocktail of alkaloids” where peramine is the main toxic substance present in endophyte of ryegrass and responsible for damage reduction of L. bonariensis. In the last decade, few studies have been developed on the effectiveness of new ryegrass cultivars carriers of endophyte in controlling insect pests. Therefore, the aim of this research is to provide knowledge concerning to evaluate the alkaloid content, such as peramine and Lolitrem B, present in new experimental lines of ryegrass and feasible to be used in grasslands of southern Chile. For this, during 2016, ryegrass plants of six experimental lines and two commercial cultivars sown at the Instituto de Investigaciones Agropecuarias Carrillanca (Vilcún, Chile) were collected and subjected to a process of chemical extraction to identify and quantify the presence of peramine and lolitrem B by the technique of liquid chromatography of high resolution (HPLC). The results indicated that the experimental lines EL-1 and EL-3 had high content of peramine (0.25 and 0.43 ppm, respectively) than with lolitrem B (0.061 and 0.19 ppm, respectively). Furthermore, the higher contents of lolitrem B were detected in the EL-4 and commercial cultivar Alto (positive control) with 0.08 and 0.17 ppm, respectively. Peramine and lolitrem B were not detected in the cultivar Jumbo (negative control). These results suggest that EL-3 would have potential as future cultivate because it has high content of peramine, alkaloid responsible for controlling insect pest. However, their current role on the complex insects attacking ryegrass grasslands should be evaluated. The information obtained in this research could be used to improve control strategies against hypogeous and epigean pests of grassland in southern Chile and also to reduce the use of synthetic pesticides.

Keywords: HPLC, Lolitrem B, peramine, pest

Procedia PDF Downloads 242

Authors: M. A. El-Khateeb

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The growth of the olive oil production in Saudi Arabia peculiarly in Al Jouf region in recent years has been accompanied by an increase in the discharge of associated processing waste. Olive mill waste is produced throughout the extraction of oil from the olive fruit using the traditional mill and press process. Deterioration of the environment due to olive mill disposal wastes is a serious problem. When olive mill waste disposed into the soil, it affects soil quality, soil micro flora, and also toxic to plants. The aim of this work is to isolate microorganism (bacterial or fungal strains) from OMW capable of degrading phenols. Olive mill wastewater, olive mill waste and soil (beside oil production mill) contaminated with olive waste were used for isolation of phenol tolerant microorganisms. Four strains (two fungal and two bacterial) were isolated from olive mill waste. The isolated strains were Candida tropicalis and Phanerochaete chrysosporium (fungal strains) and Bacillus sp. and Rhodococcus sp. (bacterial strains). These strains were able to degrade phenols and could be used for bioremediation of olive mill waste.

Keywords: bioremediation, bacteria, fungi, Sakaka

Procedia PDF Downloads 363

Authors: Cherity A. Ysquierdo, Pia U. Olafson, Donald B. Thomas

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Musca domestica L. were collected from cattle diagnosed with bovine ringworm to evaluate the potential of the house fly to disseminate Trichophyton verrucosum E. Bodin, a fungal dermatophyte that is the causative agent for ringworm in cattle. Fungal isolates were cultured from 45 individual flies on supplemented Sabouraud dextrose agar, and isolates were identified using morphological and microscopic approaches. Each isolate was further identified by PCR amplification of the ribosomal DNA locus with fungal specific primers and subsequent amplicon sequencing. No T. verrucosum were identified using these approaches. However, 36 different fungal species representing 17 genera were cultured from these flies, including several allergenic and pathogenic species. Several species within the fungal orders Hypocreales, Microascales, Onygenales, Saccharomycetales, Xylaniales, and Agaricales were observed for the first time on house flies. The most frequent fungus recovered was Cladosporium cladosporoides, which is known to be a ubiquitous, airborne allergen.

Keywords: bovine ringworm, Cladosporium, dermatophyte, Musca domestica

Procedia PDF Downloads 193