Search results for: fungal
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 347

Search results for: fungal

107 Cytotoxic Activity of Marine-derived Fungi Trichoderma Longibrachiatum Against PANC-1 Cell Lines

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

Abstract:

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

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106 Isolation and Identification Fibrinolytic Protease Endophytic Fungi from Hibiscus Leaves in Shah Alam

Authors: Mohd Sidek Ahmad, Zainon Mohd Noor, Zaidah Zainal Ariffin

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Fibrin degradation is an important part in prevention or treatment of intravascular thrombosis and cardiovascular diseases. Plasmin like fibrinolytic enzymes has given new hope to patient with cardiovascular diseases by treating fibrin aggregation related diseases with traditional plasminogen activator which have many side effects. Various researches involving wide range of sources for production of fibrinolytic proteases, from bacteria, fungi, insects and fermented foods. But few have looked into endophytic fungi as a potential source. Sixteen (16) endophytic fungi were isolated from Hibiscus sp. leaves from six different locations in Shah Alam, Selangor. Only two endophytic fungi, FH3 and S13 showed positive fibrinolytic protease activities. FH3 produced 5.78cm and S13 produced 4.48cm on Skim Milk Agar after 4 days of incubation at 27°C. Fibrinolytic activity was observed; 3.87cm and 1.82cm diameter clear zone on fibrin plate of FH3 and S13 respectively. 18srRNA was done for identification of the isolated fungi with positive fibrinolytic protease. S13 had the highest similarity (100%) to that of Penicillium citrinum strain TG2 and FH3 had the highest similarity (99%) to that of Fusarium sp. FW2PhC1, Fusarium sp. 13002, Fusarium sp. 08006, Fusarium equiseti strain Salicorn 8 and Fungal sp. FCASAn-2. Media composition variation showed the effects of carbon nitrogen on protein concentration, where the decrement of 50% of media composition caused drastic decrease in protease of FH3 from 1.081 to 0.056 and also S13 from 2.946 to 0.198.

Keywords: isolation, identification, fibrinolytic protease, endophytic fungi, Hibiscus leaves

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105 Contribution to the Production of Phenazine Antibiotics Effect Type Compounds by Some Strains of Pseudomonas spp.fluorescent

Authors: Nacéra Benoussaid, Lehalali Meriem, Benchabane Messaoud

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Our work focuses on the production of compound antibiotic effect of volatile nature namely hydrogen cyanide and the production and identification of molecules phénazinique by some strains of fluorescent Pseudomonas spp isolated from the rhizosphere of some trees for a possible use as bio pesticides antifungal effect and/or antibiotic. We tested the production of hydrogen cyanide of 21 strains of Pseudomonas spp. fluorescent among them 19 strains (90, 47%) showed a positive cyanogenesis.The antagonism test executed in vitro showed that Pseudomonas strains have a higher anti fungal effect relative to their antibacterial effect with diameters of inhibition zones up to 3, 9 cm recorded by the strain F48 against Coleosporiumsp compared with recorded results against bacteria with a maximum inhibition of 1, 26 cm among this antagonistic strain.Three strains were selected by testing for producing phénazines namely PI9, BB9 and F20. The effect of the antimicrobial activity was performed on different culture media (GN, King B, ISP2 and PDA). The results of our study allowed us to retain the King B medium as ideal medium for the production of secondary metabolite. The produced phenazinique compounds was extracted from various organic solvents, and after the results of antibiographie against germs - targets, the extracts of ethyl acetate gave the best results compared to dichloromethane and hexane.The Analysis of these compounds of antibiotic phenazinique effect within layer chromatography (CCM) and high performance liquid chromatography( HPLC) indicate that both strains PI9 and F20 are productive of phenazine-1-carboxylic acid (PCA). The BB9 strain is suspected to be productive of another phenazinique compound.

Keywords: Pseudomonas ssp. fluorescents, antagonism in vitro, secondary metabolite, phenazines, biopesticide.

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104 Assessment of Bio-Control Quality of Ethanolic Extracts of Some Tropical Plants on Fruit Rot Pathogens of Pineapple Fruits in Ado Ekiti

Authors: J. Y. Ijato, A. Adewumi, H. O Yakubu, O. O. Olajide, B. O. Ojo, B. A. Adanikin

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Post-harvest fruit rot pathogens are one of the major factors that are responsible for food security challenges in developing countries like Nigeria. These pathogens also cause fruit food poisoning. Biocidal effects of ethanolic extracts of Khaya grandifoliola, Hyptis suaveolens, Zingiber officinale, Calophyllum inophyllum, Datura stramonium on the mycelia growth of fungal rot pathogens of pineapple fruit was investigated, the ethanolic extracts of these test plants exhibited high significant inhibitory effects on the rot pathogens, the highest ethanolic extract inhibition of Zingiber officinale was on Aspergillus flavus (38.40%) at 1.0g/ml while the least inhibitory effect was on Aspergillus fumigatus (23.10%) at 1.0g/ml, the highest ethanol extract inhibition of Datura stramonium was on Aspergillus tubingensis (24.00%) at 1.0g/ml while the least inhibitory effect was 10.00% on Colletotrichum fruticola at 1.0g/ml, the highest ethanol extract inhibition of Calophyllum inophyllum was on Trichoderma harzianum (18.50%) at 1.0g/ml while the least inhibitory effect was on Aspergillus flavus (15.00%) at 1.0g/ml, the highest ethanol extract inhibition of Hyptis suaveolens was on Aspergillus fumigatus (35.00%) at 1.0g/ml while the least inhibitory effect was on Aspergillus niger (20.00%) at 1.0g/ml, the highest ethanol extract inhibition of Khaya grandifoliola was on Aspergillus flavus (35.00%) at 1.00g/ml while the least inhibitory effect was on Aspergillus fumigates (22.00%) at 1.0g/ml, the antifungal capacity of these test plant extracts on rot causing fungi on pineapple fruit reveals the possibility of their use by farmers and fruit traders as alternative to chemical fungicide that portends great threat to human and environmental health.

Keywords: fruit rot, pathogens, plant extracts, pineapple, food poisoning

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103 Production of Mycelial Biomass, Exopolysaccharide, and Enzyme during Solid-State Fermentation of Plant Raw Materials by Medicinal Mushrooms

Authors: Tamar Khardziani, Violeta Berikashvili, Amrosi Chkuaseli, Vladimir Elisashvili

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The main objectives of this proposal are to develop low-cost, innovative, and competitive technologies for the production of mycelial biomass of medicinal mushrooms as a natural food supplement for poultry. To fulfill this task, industrial strains of Lentinus edodes, Ganoderma lucidum, and Pleurotus ostreatus were used in this study. The solid-state fermentation (SSF) of wheat grains, wheat bran, and soy flour was performed in flasks and bags. Among nine mushroom strains, P. ostreatus 2191 appeared to be the most productive in protein biomass accumulation in the SSF of wheat bran. All mushrooms produced exopolysaccharide with the highest yield of 5-8 mg/mL depending on fungal strain and growth substrate. Supplementation of medium with 1% glycerol and 2-4% peptone favored mushroom growth and protein accumulation. Among inorganic nitrogen sources, KNO₃ also provided high biomass and protein production. The SSF of all growth substrates was accompanied by the secretion of cellulase and xylanase activities. The highest CMCase activity (12-13 U/g) was revealed in the cultivation of P. ostreatus 2191 using wheat bran as a growth substrate and ammonium sulfate or yeast extract as a nitrogen source, whereas the highest xylanase activity was detected in the fermentation of soy flour supplemented with peptone. Acknowledgments: This work was supported by the Shota Rustaveli National Science Foundation of Georgia (Grant number STEM-22-2077).

Keywords: mushrooms, plant raw materials, fermentation, biomass protein, cellulase

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102 Culturable Microbial Diversity of Agave Artisanal Fermentations from Central Mexico

Authors: Thalía Moreno-García Malo, Santiago Torres-Ríos, María G. González-Cruz, María M. Hernández-Arroyo, Sergio R. Trejo-Estrada

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Agave atrovirens is the main source of agave sap, the raw material for the production of pulque, an artisanal fermented beverage, traditional since prehispanic times in the highlands of central Mexico. Agave sap is rich in glucose, sucrose and fructooligosaccharides, and strongly differs from agave syrup from A. tequilana, which is mostly a high molecular weight fructan. Agave sap is converted into pulque by a highly diverse microbial community which includes bacteria, yeast and even filamentous fungi. The bacterial diversity has been recently studied. But the composition of consortia derived from directed enrichments differs sharply from the whole fermentative consortium. Using classical microbiology methods, and selective liquid and solid media formulations, either bacterial or fungal consortia were developed and analyzed. Bacterial consortia able to catabolize specific prebiotic saccharides were selected and preserved for future developments. Different media formulations, selective for bacterial genera such as Bifidobacterium, Lactobacillus, Pediococcus, Lactococcus and Enterococcus were also used. For yeast, specific media, osmotic pressure and unique carbon sources were used as selective agents. Results show that most groups are represented in the enrichment cultures; although very few are recoverable from the whole consortium in artisanal pulque. Diversity and abundance vary among consortia. Potential bacterial probiotics obtained from agave sap and agave juices show tolerance to hydrochloric acid, as well as strong antimicrobial activity.

Keywords: Agave, pulque, microbial consortia, prebiotic activity

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101 Biosynthesis, Characterization and Interplay of Bacteriocin-nanoparticles to Combat Infectious Drug Resistant Pathogens

Authors: Asma Ansari, Afsheen Aman, Shah Ali Ul Qader

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In the past few years, numerous concerns have been raised against increased bacterial resistance towards effective drugs and become a debated issue all over the world. With the emergence of drug resistant pathogens, the interaction of natural antimicrobial compounds and antibacterial nanoparticles has emerged as a potential candidate for combating infectious diseases. Microbial diversity in the biome provides an opportunity to screen new species which are capable of producing large number of antimicrobial compounds. Among these antimicrobial compounds, bacteriocins are highly specific and efficient antagonists. A combination of bacteriocin along with nanoparticles could prove to be more potent due to broadened antibacterial spectrum with possibly lower doses. In the current study, silver nanoparticles were synthesized through biological reduction using various isolated bacterial, fungal and yeast strains. Spectroscopy and scanning electron microscopy (SEM) was performed for the confirmation of nanoparticles. Bacteriocin was characterized and purified to homogeneity through gel permeation chromatography. The estimated molecular weight of bacteriocin was 10 kDa. Amino acid analysis and N-terminal sequencing revealed the novelty of the protein. Then antibacterial potential of silver nanoparticles and broad inhibitory spectrum bacteriocin was determined through agar well diffusion assay. These synthesized bacteriocin-Nanoparticles exhibit a good potential for clinical applications as compared to bacteriocin alone. This combination of bacteriocin with nanoparticles will be used as a new sort of biocide in the field of nano-proteomics. The advancement of nanoparticles-mediated drug delivery system will open a new age for rapid eradication of pathogens from biological systems.

Keywords: BAC-IB17, multidrug resistance, purification, silver nanoparticles

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100 Preliminary Studies on the Potentials of Bambara nut (Voandzeia substerranea) and Pigeon pea (Cajanus cajan) as Imitation Milk

Authors: Onuoha Gideon

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The preliminary studies on the potentials of Bambara nut and pigeon pea as imitation milk were investigated. Bambara nut and Pigeon pea milk were produced from two separate unit operations; Bambara nut seed was cooked, dehulled, milled and strained to milk (BCM) and another batch was toasted at moderate temperature, dehulled, milled and strained to milk (BTM). Pigeon pea seed was cooked, dehulled, milled and strained to milk (PCM) and another batch was toasted at moderate temperature, dehulled, milled and strained to milk (PTM). The result of the proximate analysis on the milk samples on wet basis showed that the protein content ranged from 28.56 – 26.77, the crude fibre ranged from 6.28 – 1.85, the ash content ranged from 5.22 – 1.17, the fat content ranged from 2.71 – 1.12, the moisture content ranged from 95.93 – 93.83, the carbohydrate content ranged from 67.62 – 58.83. The functional analysis on the milk samples showed that emulsification capacity ranged from 43.21 – 38.66, emulsion stability ranged from 34.10 – 25.00, the specific gravity ranged from 997.50 – 945.00, the foaming capacity ranged from 3,500 to 2,250, the measurement of viscosity ranged from 0.017 – 0.007, the pH range from 5.55 – 5.25, the measurement of dispersibility range from 11.00 – 7.00, the total soluble solid ranged from 4.00 to 1.75, the total titratable acidity ranged from 0.314 – 0.328. The sensory evaluation report showed that in terms of flavor, sample BCM and PCM value were significantly different from sample BTM and PTM. In terms of colour, sample BCM showed a significant difference from samples BTM, PCM and PTM. In term of texture, sample BCM was significantly different from samples BTM, PCM and PTM. The general acceptability shows that sample BCM was significantly different from other the samples and was the most accepted. The microbial analysis indicated that the microbial load increases with time. Bacterial count ranged from 1.3 x 105 – 1.20 x 106 to 1.6 x 105 – 1.06 x 106, fungal count ranged from 4.0 x 105 – 8.0 x 105 to 4.0 x 105 – 7.0 x 105. The studies showed that BCM was the most preferred.

Keywords: imitation milk, Bambara nut, Pigeon pea, proximate composition

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99 Beneficiation of Pulp and Paper Mill Sludge for the Generation of Single Cell Protein for Fish Farming

Authors: Lucretia Ramnath

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Fishmeal is extensively used for fish farming but is an expensive fish feed ingredient. A cheaper alternate to fishmeal is single cell protein (SCP) which can be cultivated on fermentable sugars recovered from organic waste streams such as pulp and paper mill sludge (PPMS). PPMS has a high cellulose content, thus is suitable for glucose recovery through enzymatic hydrolysis but is hampered by lignin and ash. To render PPMS amenable for enzymatic hydrolysis, the PPMS waspre-treated to produce a glucose-rich hydrolysate which served as a feed stock for the production of fungal SCP. The PPMS used in this study had the following composition: 72.77% carbohydrates, 8.6% lignin, and 18.63% ash. The pre-treatments had no significant effect on lignin composition but had a substantial effect on carbohydrate and ash content. Enzymatic hydrolysis of screened PPMS was previously optimized through response surface methodology (RSM) and 2-factorial design. The optimized protocol resulted in a hydrolysate containing 46.1 g/L of glucose, of which 86% was recovered after downstream processing by passing through a 100-mesh sieve (38 µm pore size). Vogel’s medium supplemented with 10 g/L hydrolysate successfully supported the growth of Fusarium venenatum, conducted using standard growth conditions; pH 6, 200 rpm, 2.88 g/L ammonium phosphate, 25°C. A maximum F. venenatum biomass of 45 g/L was produced with a yield coefficient of 4.67. Pulp and paper mill sludge hydrolysate contained approximately five times more glucose than what was needed for SCP production and served as a suitable carbon source. We have shown that PPMS can be successfully beneficiated for SCP production.

Keywords: pulp and paper waste, fungi, single cell protein, hydrolysate

Procedia PDF Downloads 162
98 Saponins vs Anthraquinones: Different Chemicals, Similar Ecological Roles in Marine Symbioses

Authors: Guillaume Caulier, Lola Brasseur, Patrick Flammang, Pascal Gerbaux, Igor Eeckhaut

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Saponins and quinones are two major groups of secondary metabolites widely distributed in the biosphere. More specifically, triterpenoid saponins and anthraquinones are mainly found in a wide variety of plants, bacteria and fungi. In the animal kingdom, these natural organic compounds are rare and only found in small quantities in arthropods, marine sponges and echinoderms. In this last group, triterpenoid saponins are specific to holothuroids (sea cucumbers) while anthraquinones are the chemical signature of crinoids (feather stars). Depending on the species, they present different molecular cocktails. Despite presenting different chemical properties, these molecules share numerous similarities. This study compares the biological distribution, the pharmacological effects and the ecological roles of holothuroid saponins and crinoid anthraquinones. Both of them have been defined as allomones repelling predators and parasites (i.e. chemical defense) and have interesting pharmacological properties (e.g. anti-bacterial, anti-fungal, anti-cancer). Our study investigates the chemical ecology of two symbiotic associations models; between the snapping shrimp Synalpheus stimpsonii associated with crinoids and the Harlequin crab Lissocarcinus orbicularis associated with holothuroids. Using behavioral experiments in olfactometers, chemical extractions and mass spectrometry analyses, we discovered that saponins and anthraquinones present a second ecological role: the attraction of obligatory symbionts towards their hosts. They can, therefore, be defined as kairomones. This highlights a new paradigm in marine chemical ecology: Chemical repellents are attractants to obligatory symbionts because they constitute host specific chemical signatures.

Keywords: anthraquinones, kairomones, marine symbiosis, saponins, attractant

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97 Genome-Wide Identification and Characterization of MLO Family Genes in Pumpkin (Cucurbita maxima Duch.)

Authors: Khin Thanda Win, Chunying Zhang, Sanghyeob Lee

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Mildew resistance locus o (Mlo), a plant-specific gene family with seven-transmembrane (TM), plays an important role in plant resistance to powdery mildew (PM). PM caused by Podosphaera xanthii is a widespread plant disease and probably represents the major fungal threat for many Cucurbits. The recent Cucurbita maxima genome sequence data provides an opportunity to identify and characterize the MLO gene family in this species. Total twenty genes (designated CmaMLO1 through CmaMLO20) have been identified by using an in silico cloning method with the MLO gene sequences of Cucumis sativus, Cucumis melo, Citrullus lanatus and Cucurbita pepo as probes. These CmaMLOs were evenly distributed on 15 chromosomes of 20 C. maxima chromosomes without any obvious clustering. Multiple sequence alignment showed that the common structural features of MLO gene family, such as TM domains, a calmodulin-binding domain and 30 important amino acid residues for MLO function, were well conserved. Phylogenetic analysis of the CmaMLO genes and other plant species reveals seven different clades (I through VII) and only clade IV is specific to monocots (rice, barley, and wheat). Phylogenetic and structural analyses provided preliminary evidence that five genes belonged to clade V could be the susceptibility genes which may play the importance role in PM resistance. This study is the first comprehensive report on MLO genes in C. maxima to our knowledge. These findings will facilitate the functional analysis of the MLOs related to PM susceptibility and are valuable resources for the development of disease resistance in pumpkin.

Keywords: Mildew resistance locus o (Mlo), powdery mildew, phylogenetic relationship, susceptibility genes

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96 An Advanced Method of Plant Preservation and Colour Retention of Herbarium Specimens

Authors: Abduraheem K., Suboohi Nasrin

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Herbaria are specimens of preserved plants, which are very delicate and cellulosic in nature. While these collections are very useful for the enrichment of knowledge and are considered as natural heritage of our entire world, it is very important to preserve and conserve them. The significance is not only to prevent the herbaria from the deterioration of biological agencies but also to preserve its colours and retain natural colour. Colour is not only characteristic of a plant, but it can also help to identify closely related species or to distinguish a plant from a collection of herbaria. Keeping this in mind, a selective solution has been prepared for the conservation and preservation of herbarium in the present study. In this, the quantity of all the selected chemicals, i.e., formaldehyde and copper sulphate was kept constant, and the solution was prepared by dissolving it in distilled water by increasing the amount of picric acid (1, 2, 3, 4, and 5 ml). Fresh specimens of roses and bougainvillea were washed with distilled water and kept in the above solution for 10 to 15 minutes at room temperature. After 10 minutes, the specimen was removed from the solution, dried with the help of paper, and then pressed under the plant press. Blotting sheets were used to absorb the moisture content and were changed every 2 to 3 days to protect against fungal growth. The results revealed that all solutions had insecticidal properties and protected the herbarium specimen against pests. While in the case of colour retention, solution-1 and 2 were not satisfactory colour preservation, and solutions-3 and 5 maintained the colour of rose and bougainvillea leaves for 15 to 20 days and for a month, respectively. After that, the colour begins to fade, and the process is faster in rose leaves than in bougainvillea. And it was also observed that the colour of young leaves started to fade before that of older leaves. When the leaves of rose and bougainvillea are treated with Solution-4, then the colour of rose leaves is maintained for six months.

Keywords: solutions, colour retention, preservation and conservation, leaves of roses and bougainvillea

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95 Rhizosphere Microbiome Involvement in the Natural Suppression of Soybean Cyst Nematode in Disease Suppressive Soil

Authors: M. Imran Hamid, Muzammil Hussain, Yunpeng Wu, Meichun Xiang, Xingzhong Liu

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The rhizosphere microbiome elucidate multiple functioning in the soil suppressiveness against plant pathogens. Soybean rhizosphere microbial communities may involve in the natural suppression of soybean cyst nematode (SCN) populations in disease suppressive soils. To explore these ecological mechanisms of microbes, a long term monoculture suppressive soil were taken into account for further investigation to test the disease suppressive ability by using different treatments. The designed treatments are as, i) suppressive soil (S), ii) conducive soil (C), iii) conducive soil mixed with 10% (w/w) suppressive soil (CS), iv) suppressive soil treated at 80°C for 1 hr (S80), and v) suppressive soil treated with formalin (SF). By using an ultra-high-throughput sequencing approach, we identified the key bacterial and fungal taxa involved in SCN suppression. The Phylum-level investigation of bacteria revealed that Actinobacteria, Bacteroidetes, and Proteobacteria in the rhizosphere soil of soybean seedlings were more abundant in the suppressive soil than in the conducive soil. The phylum-level analysis of fungi in rhizosphere soil indicated that relative abundance of Ascomycota was higher in suppressive soil than in the conducive soil, where Basidiomycota was more abundant. Transferring suppressive soil to conducive soil increased the population of Ascomycota in the conducive soil by lowering the populations of Basidiomycota. The genera, such as, Pochonia, Purpureocillium, Fusarium, Stachybotrys that have been well documented as bio-control agents of plant nematodes were far more in the disease suppressive soils. Our results suggested that the plants engage a subset of functional microbial groups in the rhizosphere for initial defense upon nematode attack and protect the plant roots later on by nematodes to response for suppression of SCN in disease-suppressive soils.

Keywords: disease suppressive soil, high-throughput sequencing, rhizosphere microbiome, soybean cyst nematode

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94 Antimicrobial Activity of Biosynthesized Silver Nanoparticles Using Different Bacteria

Authors: Malalage Mudara Peiris

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Objectives of the study are: the biosynthesis of silver nanoparticles (AgNPs) using Escherichia coli, Acinetobacter baumannii and Staphylococcus aureus, characterization of silver nanoparticles and determination of antimicrobial activity against E. coli, P. aeruginosa, S. aureus, MRSA, and C. Albicans. Methods: E. coli (ATCC 25922), A. baumanii (clinical strain), S. aureus (clinical strain) cultured in nutrient broth medium were used for biosynthesis of AgNPs. Culture conditions (AgNO3 concentration, pH, incubation time and temperature) were optimized. Characterization of synthesized NPs was done by UV-Visible spectroscopy. The antimicrobial activity of the synthesized NPs was studied using the good diffusion assay against E. coli, S. aureus, MRSA (Methicillin-resistant Staphylococcus aureus), P. aeruginosa and C. Albicans. Results: All the selected bacteria produced silver nanoparticles at alkaline pH above 0.3 g/L AgNO3 concentration. The optimum reaction temperature was 60oC. According to the UV-Visible spectroscopy, the maximum absorbance was found to be around 420 - 430 nm indicating the presence of AgNPs. According to the good diffusion results, AgNPs produced by S. aureus resulted in the larger zone of inhibition (ZOI) against the selected pathogens, while AgNPs produced by E. coli showed comparatively smaller ZOI. In general, biosynthesized AgNPs were highly effective against gram-negative bacteria compared to gram-positive bacterial and fungal species. Conclusions: Green AgNPs produced by each bacterium show antimicrobial activity against the selected pathogens. AgNPs produced by S. aureus are the most effective NPs among tested AgNPs, while AgNPs produced by E. coli are the least effective. Further characterization of NPs is required to study the physical properties of silver NPs.

Keywords: green nanotechnology, silver nanoparticles, bacteria, antimicrobial activity

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93 Combined Use of Microbial Consortia for the Enhanced Degradation of Type-IIx Pyrethroids

Authors: Parminder Kaur, Chandrajit B. Majumder

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The unrestrained usage of pesticides to meet the burgeoning demand of enhanced crop productivity has led to the serious contamination of both terrestrial and aquatic ecosystem. The remediation of mixture of pesticides is a challenging affair regarding inadvertent mixture of pesticides from agricultural lands treated with various compounds. Global concerns about the excessive use of pesticides have driven the need to develop more effective and safer alternatives for their remediation. We focused our work on the microbial degradation of a mixture of three Type II-pyrethroids, namely Cypermethrin, Cyhalothrin and Deltamethrin commonly applied for both agricultural and domestic purposes. The fungal strains (Fusarium strain 8-11P and Fusarium sp. zzz1124) had previously been isolated from agricultural soils and their ability to biotransform this amalgam was studied. In brief, the experiment was conducted in two growth systems (added carbon and carbon-free) enriched with variable concentrations of pyrethroids between 100 to 300 mgL⁻¹. Parameter optimization (pH, temperature, concentration and time) was done using a central composite design matrix of Response Surface Methodology (RSM). At concentrations below 200 mgL⁻¹, complete removal was observed; however, degradation of 95.6%/97.4 and 92.27%/95.65% (in carbon-free/added carbon) was observed for 250 and 300 mgL⁻¹ respectively. The consortium has been shown to degrade the pyrethroid mixture (300 mg L⁻¹) within 120 h. After 5 day incubation, the residual pyrethroids concentration in unsterilized soil were much lower than in sterilized soil, indicating that microbial degradation predominates in pyrethroids elimination with the half-life (t₁/₂) of 1.6 d and R² ranging from 0.992-0.999. Overall, these results showed that microbial consortia might be more efficient than single degrader strains. The findings will complement our current understanding of the bioremediation of mixture of Type II pyrethroids with microbial consortia and potentially heighten the importance for considering bioremediation as an effective alternative for the remediation of such pollutants.

Keywords: bioremediation, fungi, pyrethroids, soil

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92 Allele Mining for Rice Sheath Blight Resistance by Whole-Genome Association Mapping in a Tail-End Population

Authors: Naoki Yamamoto, Hidenobu Ozaki, Taiichiro Ookawa, Youming Liu, Kazunori Okada, Aiping Zheng

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Rice sheath blight is one of the destructive fungal diseases in rice. We have thought that rice sheath blight resistance is a polygenic trait. Host-pathogen interactions and secondary metabolites such as lignin and phytoalexins are likely to be involved in defense against R. solani. However, to our knowledge, it is still unknown how sheath blight resistance can be enhanced in rice breeding. To seek for an alternative genetic factor that contribute to sheath blight resistance, we mined relevant allelic variations from rice core collections created in Japan. Based on disease lesion length on detached leaf sheath, we selected 30 varieties of the top tail-end and the bottom tail-end, respectively, from the core collections to perform genome-wide association mapping. Re-sequencing reads for these varieties were used for calling single nucleotide polymorphisms among the 60 varieties to create a SNP panel, which contained 1,137,131 homozygous variant sites after filitering. Association mapping highlighted a locus on the long arm of chromosome 11, which is co-localized with three sheath blight QTLs, qShB11-2-TX, qShB11, and qSBR-11-2. Based on the localization of the trait-associated alleles, we identified an ankyryn repeat-containing protein gene (ANK-M) as an uncharacterized candidate factor for rice sheath blight resistance. Allelic distributions for ANK-M in the whole rice population supported the reliability of trait-allele associations. Gene expression characteristics were checked to evaluiate the functionality of ANK-M. Since an ANK-M homolog (OsPIANK1) in rice seems a basal defense regulator against rice blast and bacterial leaf blight, ANK-M may also play a role in the rice immune system.

Keywords: allele mining, GWAS, QTL, rice sheath blight

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91 A High Content Screening Platform for the Accurate Prediction of Nephrotoxicity

Authors: Sijing Xiong, Ran Su, Lit-Hsin Loo, Daniele Zink

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The kidney is a major target for toxic effects of drugs, industrial and environmental chemicals and other compounds. Typically, nephrotoxicity is detected late during drug development, and regulatory animal models could not solve this problem. Validated or accepted in silico or in vitro methods for the prediction of nephrotoxicity are not available. We have established the first and currently only pre-validated in vitro models for the accurate prediction of nephrotoxicity in humans and the first predictive platforms based on renal cells derived from human pluripotent stem cells. In order to further improve the efficiency of our predictive models, we recently developed a high content screening (HCS) platform. This platform employed automated imaging in combination with automated quantitative phenotypic profiling and machine learning methods. 129 image-based phenotypic features were analyzed with respect to their predictive performance in combination with 44 compounds with different chemical structures that included drugs, environmental and industrial chemicals and herbal and fungal compounds. The nephrotoxicity of these compounds in humans is well characterized. A combination of chromatin and cytoskeletal features resulted in high predictivity with respect to nephrotoxicity in humans. Test balanced accuracies of 82% or 89% were obtained with human primary or immortalized renal proximal tubular cells, respectively. Furthermore, our results revealed that a DNA damage response is commonly induced by different PTC-toxicants with diverse chemical structures and injury mechanisms. Together, the results show that the automated HCS platform allows efficient and accurate nephrotoxicity prediction for compounds with diverse chemical structures.

Keywords: high content screening, in vitro models, nephrotoxicity, toxicity prediction

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90 Improving the Feeding Value of Straws with Pleurotus Ostreatus

Authors: S. Hussain, N. Ahmad, S. Alam, M. Bezabhi, W. H. Hendriks, P. Yu, J. W. Cone

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The high content of lignin in cell walls is the major limiting factor in the digestion and utilisation of cereal crop residues by ruminants. The aim of this study was to evaluate the effectiveness of the white rot fungus, Pleurotus ostreatus (P. ostreatus), to degrade lignin and to enhance the rumen degradability of maize stover, rice straw, wheat straw and their mixture in equal proportion on a dry-matter (DM) basis. Four samples of each substrate were incubated aerobically in triplicate with P. ostreatus for 0 (Control), 21, 28 and 35 days under solid-state conditions (temperature, 24 ͦ C; humidity, 70± 5%). The changes in chemical composition, DM and nutrient losses, and rumen fermentation characteristics using in vitro DM digestibility (DMD) and the in vitro gas production (GP) technique were measured. The results showed that incubation with P. ostreatus decreased (P < 0.001) the contents of neutral detergent fibre and lignin with a concomitant increase (P < 0.001) in the contents of ash and crude protein. The losses of nutrients differed (P < 0.001) among the straw types, with rice straw and maize stover showing the largest (P < 0.05) lignin degradation compared to wheat and mixed straws. The DMD and 72-h cumulative GP increased (P < 0.001) consistently with increasing fungal incubation period and for all substrates the highest values of DMD and GP were measured after 35 days of incubation with P. ostreatus. The lignin degradation was strongly associated with hemicellulose degradation (r = 0.71) across the various straws. Results of the present study demonstrated that incubation of low-quality crop residues with P. ostreatus under solid-state conditions upgrades their feeding value by reducing the content of lignin and increasing the content of crude protein and ruminal degradation.

Keywords: crop residues, lignin degradation, maize stovers, wheat straws, white rot fungi

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89 Fungi Associated with Decline of Kikar (Acacia nilotica) and Red River Gum (Eucalyptus camaldulensis) in Faisalabad

Authors: I. Ahmad, A. Hannan, S. Ahmad, M. Asif, M. F. Nawaz, M. A. Tanvir, M. F. Azhar

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During this research, a comprehensive survey of tree growing areas of Faisalabad district of Pakistan was conducted to observe the symptoms, spectrum, occurrence and severity of A. nilotica and E. camaldulensis decline. Objective of current research was to investigate specific fungal pathogens involved in decline of A. nilotica and E. camaldulensis. For this purpose, infected roots, bark, neck portion, stem, branches, leaves and infected soils were collected to identify associated fungi. Potato dextrose agar (PDA) and Czepak dox agar media were used for isolations. Identification of isolated fungi was done microscopically and different fungi were identified. During survey of urban locations of Faisalabad, disease incidence on Kikar and Eucalyptus was recorded as 3.9-7.9% and 2.6-7.1% respectively. Survey of Agroforest zones of Faisalabad revealed decline incidence on kikar 7.5% from Sargodha road while on Satiana and Jhang road it was not planted. In eucalyptus trees, 4%, 8% and 0% disease incidence was observed on Jhang road, Sargodha road and Satiana road respectively. The maximum fungus isolated from the kikar tree was Drechslera australiensis (5.00%) from the stem part. Aspergillus flavus also gave the maximum value of (3.05%) from the bark. Alternaria alternata gave the maximum value of (2.05%) from leaves. Rhizopus and Mucor spp. were recorded minimum as compared to the Drechslera, Alternaria and Aspergillus. The maximum fungus isolated from the Eucalyptus tree was Armillaria luteobubalina (5.00%) from the stem part. The other fungi isolated were Macrophamina phaseolina and A. niger.

Keywords: decline, frequency of mycoflora, A. nilotica and E. camaldulensis, Drechslera australiensis, Armillaria luteobubalina

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88 Detection and Expression of Peroxidase Genes in Trichoderma harzianum KY488466 and Its Response to Crude Oil Degradation

Authors: Michael Dare Asemoloye, Segun Gbolagade Jonathan, Rafiq Ahmad, Odunayo Joseph Olawuyi, D. O. Adejoye

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Fungi have potentials for degrading hydrocarbons through the secretion of different enzymes. Crude oil tolerance and degradation by Trichoderma harzianum was investigated in this study with its ability to produce peroxidase enzymes (LiP and MnP). Many fungal strains were isolated from rhizosphere of grasses growing on a crude oil spilled site, and the most frequent strain based on percentage incidence was further characterized using morphological and molecular characteristics. Molecular characterization was done through the amplification of Ribosomal-RNA regions of 18s (1609-1627) and 28s (287-266) using ITS1 and ITS4 combinations and it was identified using NCBI BLAST tool. The selected fungus was also subjected to an in-vitro tolerance test at crude oil concentrations of 5, 10, 15, 20 and 25% while 0% served as control. In addition, lignin peroxidase genes (lig1-6) and manganese peroxidase gene (mnp) were detected and expressed in this strain using RT-PCR technique, its peroxidase producing activities was also studied in aliquots (U/ml). This strain had highest incidence of 80%, it was registered in NCBI as Trichoderma harzianum asemoJ KY488466. The strain KY488466 responded to crude oil concentrations as it increase, the dose inhibition response percentage (DIRP) increased from 41.67 to 95.41 at 5 to 25 % crude oil concentrations. All the peroxidase genes are present in KY488466, and expressed with amplified 900-1000 bp through RT-PCR technique. In this strain, lig2, lig4 and mnp genes were over-expressed, lig 6 was moderately expressed, while none of the genes was under-expressed. The strain also produced 90±0.87 U/ml lignin peroxidase and 120±1.23 U/mil manganese peroxidase enzymes in aliquots. These results imply that KY488466 can tolerate and survive high crude oil concentration and could be exploited for bioremediation of oil-spilled soils, the produced peroxidase enzymes could also be exploited for other biotechnological experiments.

Keywords: crude oil, enzymes, expression, peroxidase genes, tolerance, Trichoderma harzianum

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87 Distribution Frequency, Ecology, and Economic Utility of Coprophilous Mushrooms (Agaricales, Basidiomycota) in Punjab, India

Authors: Amandeep Kaur, N. S. Atri, Munruchi Kaur

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Herbivorous dung is a special substrate for the growth of fungi. Fungi growing thereon are known as coprophilous. These fungi are amongst the most abundant taxa in the ecosystem, which regulate the decomposition of dung organic matter, nutrient dynamics and maintenance of ecological balance on the earth. The coprophilous fungi represent a diverse group of saprobes, including taxa from most major fungal groups belonging to Zygomycota, Ascomycota and Basidiomycota. The present work, however, has been focused on the basidiomycetous coprophilous mushrooms belonging to the order Agaricales. The research work includes the results of eco-taxonomic studies of coprophilous mushrooms in Punjab, India, on the basis of a survey of dung localities of the state. The mushrooms were collected growing as saprobes on dung of various domesticated and wild herbivorous animals in pastures, grasslands, zoos, and on dung heaps in villages, etc. The present study observed the frequency of distribution of coprophilous mushrooms in different taxonomic categories in different regions of the state in various seasons on different dung types along with their growing habit. The paper also discusses their economic utility as edible, inedible, poisonous, medicinal and hallucinogenic species. The study has shown that animal dung is a good niche for the growth of mushrooms. However, the natural habitats with dung deposits are getting destroyed because of different developmental activities. Livestock in agriculture-based societies like Punjab state in India should be managed in a manner that favors their grazing in the wild places and thereby the growth of coprophilous mushrooms so that a significant role in ecological balance on the earth is established.

Keywords: herbivorous dung, psychoactive, seasonal availability, taxo-ecology

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86 Investigation of the Effect of Anaerobic Digestate on Antifungal Activity and in Different Parameters of Maize

Authors: Nazia Zaffar, Alam Khan, Abdul Haq, Malik Badshah

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Pakistan is an agricultural country. The increasing population leads to an increase in demand for food. A large number of crops are infected by different microbes, and nutrient deficiency of soil adversely affects the yield of crops. Furthermore, the use of chemical fertilizers like Nitrogen, Phosphorus, Potassium (NPK) Urea, and Diammonium phosphate (DAP) and pesticides have environmental consequences. Therefore, there is an urgent need to explore alternative renewable and sustainable biofertilizers. Maize is one of the top growing crops in Pakistan, but it has low yield compared to other countries due to deficiency of organic matter, widespread nutrients deficiency (phosphorus and nitrogen), unbalanced use of fertilizers and various fungal diseases. In order to get rid of all these disadvantages, Digestate emerged as a win-win opportunity for the control of a few plant diseases and a replacement for the chemical fertilizers. The present study was designed to investigate the effect of Anerobic digestate on Antifungal Activity and in different parameters of Maize. The antifungal activity, minimum inhibitory concentration (MIC), and minimum fungicidal concentration (MFC) against selected phytopathogens (Colletotrichum coccodis, Pythium ultimum, Phytophthora capsci, Rhizoctonia solani, Bipolaris oryzae and Fusarium Fujikuroi) were determined by microtiter plate method. The effect of various fertilizers in different growth parameters height, diameter, chlorophyll, leaf area, biomass, and yield were studied in field experiments. The extracts from anaerobic digestate have shown antifungal activity against selected phytopathogens, the highest activity was noted against P. ultimum, the MIC activity was high in case of P. ultimum and B. oryzae. The present study concludes that anaerobic digestate have a positive effect on maize growth and yield as well as an antifungal activity which can be potentially a good biofertilizer.

Keywords: anaerobic digestate, antifungal activity, MIC, phytopathogens

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85 Seed Dressing and Foliar Spray of Green Bean (Phaseolus vulgaris L.) with Essential Oils and Disinfectants for Suppressing Root Rot and Wilt Incidence under Field Conditions

Authors: N. S. El-Mougy, M. M. Abdel-Kader, H. M. Abouelnasr

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The efficacy of two essential oils applied as a bean seed dressing followed by seedlings foliar spray with four commercial disinfectants against root rot and wilt incidence was evaluated under field conditions. The essential oils, grape seed and peppermint oils and the disinfectants, Gold, Revarest, Klenva, Malva were applied. Chitosan and the fungicide Topsin-M were used as comparison treatment. Essential oils individually or combined with disinfectants were applied as a bean seed dressing. Furthermore, emerged bean plants were sprayed with the same treatments. Under laboratory conditions, growth inhibition effect was observed for the isolated, tested fungi R. solani and F. oxysporum when exposed to essential oils individually or combined with disinfectants. A high inhibitor effect was recorded for peppermint followed by grape seed oils. Concentrations of 1% and 2% of chitosan as well as Topsin M at 400 ppm showed complete reduction (100%) in the two fungal growths. Under field conditions, the obtained results showed that the applied treatments of chitosan had a superior effect on root rot and wilt disease incidence compared with other tested treatments. It was found that seed coating treatment provides good protection of emerged green bean seeds against the root pathogens attack compared with the fungicide and control treatments. Also, the application of seed dressing with essential oils accompanied by seedling spray demonstrated similar results. It was observed that essential oils had an enhancing effect against disease incidence when combined with disinfectants compared with their application. The obvious yield increase was significantly higher in all applied treatments than in fungicide and control.

Keywords: bean, disinfectants, essential oils, root rot, wilt

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84 In Vitro Effects of Azadirachta indica Leaves Extract Against Albugo Candida, the Causative Agent of White Blisters Disease of Brassica Oleraceae L., Var. Italica

Authors: Affiah D. U., Katuri I. P., Emefiene M. E., Amienyo C. A.

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Broccoli (Brassica oleraceae L., var. italica) is one of the most important vegetables that is high in nutrients and bioactive compounds. It easily grown on a wide range of soil types and is adaptable to many different climatic conditions. This study was carried out within Jos North and environs in vitro to evaluate Neem (Azadirachta indica) leaves extract against Albugo candida, the causative agent of white blisters disease of broccoli. Through the survey, prevalence and incidence were accessed and a fluffy white growth symptom on the underside of leaves was also observed on the field. Infected leaves samples were collected from three different farms namely: Farin Gada, Naraguta, and Juth and the organism associated with the disease was isolated. Pathogenicity test carried out revealed the fungal isolate Albugo candida to be responsible for the disease. Antimicrobial susceptibility test was performed using agar well diffusion method to determine the minimum inhibitory concentrations of two extract of Azadirachta indica leaves against the organism. Ethanolic extract had the highest antifungal activities of 3.30±0.21 - 17.61± 0.11 while aqueous extract had the least antifungal activities of 0.00±0.00 - 13.23±0.12. The minimum inhibitory concentration of aqueous was 100 mg/ml while its minimum fungicidal concentration was at 200 mg/ml. For ethanol, the minimum inhibitory concentration was 50 mg/ml while its minimum fungicidal concentration was 100 mg/ml. Plants being less toxic in usage over synthetic or inorganic chemicals makes them easy to handle, easily accessible and renewable. Due to the biosafety of plant extracts and its availability since the plant-based extracts of the two different solvents were found to be effective against the test organism hence, it is recommended for in-depth research to make it readily available for control of other pathogens and pests.

Keywords: antifungal, biocontrol, broccoli, fungi

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83 Response of Post-harvest Treatments on Shelf Life, Biochemical and Microbial Quality of Banana Variety Red Banana

Authors: Karishma Sebastian, Pavethra A., Manjula B. S., K. N. Satheeshan, Jenita Thinakaran

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Red Banana is a popular variety of banana with strong market demand. Its ripe fruits are less resistant to transportation, complicating logistics. Moreover, as it is a climacteric fruit, its post-harvest shelf life is limited. The current study aimed to increase the postharvest shelf life of Red Banana fruits by adopting different postharvest treatments. Fruit bunches of Red Banana were harvested at the mature green stage, separated into hands, precooled, subjected to 12 treatments, and stored in Corrugated Fibre Board boxes till the end of shelf life under ambient conditions. Fruits coated with 10% bee wax + 0.5% clove oil (T₄), fruits subjected to coating with 10% bee wax and packaging with potassium permanganate (T₉), and fruits dipped in hot water at 50°C for 10 minutes and packaging with potassium permanganate (T₁₁) registered the highest shelf life of 18.67 days. The highest TSS of 26.33°Brix was noticed in fruits stored with potassium permanganate (T₈) after 12.67 days of storage, and lowest titratable acidity of 0.19%, and the highest sugar-acid ratio of 79.76 was noticed in control (T₁₂) after 11.33 days of storage. Moreover, the highest vitamin C content (7.74 mg 100 g⁻¹), total sugar content (18.47%), reducing sugar content (15.49%), total carotenoid content (24.13 µg 100 g-¹) was noticed in treatments T₇ (hot water dipping at 50 °C for 10 minutes) after 17.67 days, T₁₀ (coating with 40% aloe vera extract and packaged with potassium permanganate) after 13.33 days, T₄ (coating with 10% bee wax + 0.5% clove oil) after 18.67 days and T₉ (coating with 10% bee wax + potassium permanganate) after 18.67 days of storage respectively. Furthermore, the lowest fungal and bacterial counts were observed in treatments T₂ (dipping in 30ppm sodium hypochlorite solution), T₇ (hot water dipping at 50 °C for 10 minutes), T₉ (coating with 10% bee wax + potassium permanganate), and T₁₀ (coating with 40% aloe vera extract + potassium permanganate).

Keywords: bee wax, post-harvest treatments, potassium permanganate, Red Banana, shelf life

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82 Serum MicroRNA and Inflammatory Mediators: Diagnostic Biomarkers for Endometritis in Arabian Mares

Authors: Sally Ibrahim, Mohamed Hedia, Mohamed Taqi, Mohamed Derbala, Karima Mahmoud, Youssef Ahmed, Sayed Ismail, Mohamed El-Belely

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The identification and quantification of serum microRNA (miRNA) from mares with endometritis might serve as useful and implementable clinical biomarkers for the early diagnosis of endometiritis. Aims of the current study were (I) to study the expression pattern of eca-miR-155, eca-miR-223, eca-miR-17, eca-miR-200a, and eca-miR-205, and (II) to determine the levels of interleukin 6 (IL-6), prostaglandins (PGF₂α and PGE₂), in the serum of Arabian mares with healthy and abnormal uterine status (endometritis). This study was conducted on 80 Arabian mares (4-14 years old). Mares were divided into 48 sub-fertile mares suspected of endometritis and 32 fertile at stud farms. The criteria for mares to be enrolled in the endometritis group were that they had been bred three or more times unsuccessfully in the breeding season or had a history of more than one year of reproductive failure. In addition, two or more of the following criteria on a checklist were present: abnormal clinical findings, transrectal ultrasonographic uterine examination showed abnormal fluid in the uterus (echogenic or ≥2 cm in diameter), positive endometrial cytology; and bacterial and/or fungal growth. Serum samples were collected for measuring IL-6, PGF₂α, and PGE₂ concentrations, as well as serum miRNA isolation and quantitative real-time PCR. Serum concentrations of IL-6, PGE₂, and PGF₂α were higher (P ≤ 0.001) in mares with endometritis compared to the control healthy ones. The expression profile of eca-miR-155, eca-miR-223, eca-miR-17, eca-miR-200a, and eca-miR-205 increased (P≤0.001) in mares with endometritis compared to the control ones. To the best of our knowledge, this is the first study that revealed that serum miRNA and serum inflammatory mediators (IL-6, PGE₂, and PGF₂α) could be used as non-invasive gold standard biomarkers, and therefore might be served as an important additional diagnostic tool for endometritis in Arabian mares. Moreover, estimation of the serum concentrations of serum miRNA, IL-6, PGE₂, and PGF₂α is a promising recommended tool during the breeding soundness examination in mares.

Keywords: Arabian Mares, endometritis, inflammatory mediators, serum miRNA

Procedia PDF Downloads 139
81 Production of Vermiwash from Medicinal Plants and Its Potential Use as Fungicide against the Alternaria Alternata (fr.) Keissl. Affecting Cucumber (Cucumis sativus L.) in Guyana

Authors: Abdullah Ansari, Sinika Rambaran, Sirpaul Jaikishun

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Vermiwash could be used to enhance plant productivity and resistance to some harmful plant pathogens, as well as provide benefit through the disposal of waste matter. Alternaria rot caused by the fungus Alternaria alternata (Fr.) Keissl., is a common soil-borne pathogen that results in postharvest fruit rot of cucumbers, peppers and other cash crops. The production and distribution of Cucumis sativus L. (cucumber) could be severely affected by Alternaria rot. Fungicides are the traditional treatment however; they are not only expensive but can also cause environmental and health problems. Vermiwash was prepared from various medicinal plants (Ocimum tenuiflorum L. {Tulsi}, Azadirachta indica A. Juss. {neem}, Cymbopogon citratus (DC. ex Nees) Stapf. {lemon grass} and Oryza sativa L. {paddy straw} and applied, in vitro, to A. alternata to investigate their effectiveness as organic alternatives to traditional fungicides. All of the samples of vermiwash inhibited the growth of A. alternata. The inhibitive effects on the fungus appeared most effective when A. indica and O. tenuiflorum were used in the production of the vermiwash. Using the serial dilution method, vermiwash from O. tenuiflorum showed the highest percent of inhibition (93.2%), followed by C. citratus (74.7%), A. indica (68.7%), O. sativa, combination, and combination without worms. Using the sterile disc diffusion method, all of the samples produced zones of inhibition against A. alternata. Vermiwash from A. indica produced a zone of inhibition, averaging 15.3mm, followed by O. tenuiflorum (14.0mm), combination without worms, combination, C. citratus and O. sativa. Nystatin produced a zone of inhibition of 10mm. The results indicate that vermiwash is not simply an organic alternative to more traditional chemical fungicides, but it may in fact be a better and more effective product in treating certain fungal plant infections, particularly A. alternata.

Keywords: vermiwash, earthworms, soil, bacteria, alternaria alternata, antifungal, antibacterial

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80 Investigating the Potential Use of Unsaturated Fatty Acids as Antifungal Crop Protective Agents

Authors: Azadeh Yasari, Michael Ganzle, Stephen Strelkov, Nuanyi Liang, Jonathan Curtis, Nat N. V. Kav

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Pathogenic fungi cause significant yield losses and quality reductions to major crops including wheat, canola, and barley. Toxic metabolites produced by phytopathogenic fungi also pose significant risks to animal and human health. Extensive application of synthetic fungicides is not a sustainable solution since it poses risks to human, animal and environmental health. Unsaturated fatty acids may provide an environmentally friendly alternative because of their direct antifungal activity against phytopathogens as well as through the stimulation of plant defense pathways. The present study assessed the in vitro and in vivo efficacy of two hydroxy fatty acids, coriolic acid and ricinoleic acid, against the phytopathogens Fusarium graminearum, Pyrenophora tritici-repentis, Pyrenophora teres f. teres, Sclerotinia sclerotiorum, and Leptosphaeria maculans. Antifungal activity of coriolic acid and ricinoleic acid was evaluated using broth micro-dilution method to determine the minimum inhibitory concentration (MIC). Results indicated that both ricinoleic acid and coriolic acid showed antifungal activity against phytopathogens, with the strongest inhibitory activity against L. maculans, but the MIC varied greatly between species. An antifungal effect was observed for coriolic acid in vivo against pathogenic fungi of wheat and barley. This effect was not correlated to the in vitro activity because ricinoleic acid with equivalent in vitro antifungal activity showed no protective effect in vivo. Moreover, neither coriolic acid nor ricinoleic acid controlled fungal pathogens of canola. In conclusion, coriolic acid inhibits some phytopathogens in vivo and may have the potential to be an effective crop protection agent.

Keywords: coriolic acid, minimum inhibitory concentration, pathogenic fungi, ricinoleic acid

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79 Biological Methods to Control Parasitic Weed Phelipanche ramosa L. Pomel in the Field Tomato Crop

Authors: F. Lops, G. Disciglio, A. Carlucci, G. Gatta, L. Frabboni, A. Tarantino, E. Tarantino

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Phelipanche ramosa L. Pomel is a root holoparasitic weed plant of many cultivations, particularly of tomato (Lycopersicum esculentum L.) crop. In Italy, Phelipanche problem is increasing, both in density and in acreage. The biological control of this parasitic weed involves the use of living organisms as numerous fungi and bacteria that can infect the parasitic weed, while it may improve the crop growth. This paper deals with the biocontrol with microorganism, including Arbuscular mycorrhizal (AM) fungi and fungal pathogens as Fusarium oxisporum spp. Colonization of crop roots by AM fungi can provide protection of crops against parasitic weeds because of a reduction in their seed germination and attachment, while F. oxisporum, isolated from diseased broomrape tubercles, proved to be highly virulent on P. ramosa. The experimental trial was carried out in open field at Foggia province (Apulia Region, Southern Italy), during the spring-summer season 2016, in order to evaluate the effect of four biological treatments: AM fungi and Fusarium oxisporum applied in the soil alone or combined together, and Rizosum Max® product, compared with the untreated control, to reduce the P. ramosa infestation in processing tomato crop. The principal results to be drawn from this study under field condition, in contrast of those reported previously under laboratory and greenhouse conditions, show that both AM fungi and F. oxisporum do not provide the reduction of the number of emerged shoots of P. ramosa. This can arise probably from the low efficacy seedling of the agent pathogens for the control of this parasite in the field. On the contrary, the Rizosum Max® product, containing AM fungi and some rizophere bacteria combined with several minerals and organic substances, appears to be most effective for the reduction of P. ramosa infestation.

Keywords: Arbuscular mycorrhized fungi, biocontrol methods, Phelipanche ramosa, tomato crop

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78 Carbon Sequestration and Carbon Stock Potential of Major Forest Types in the Foot Hills of Nilgiri Biosphere Reserve, India

Authors: B. Palanikumaran, N. Kanagaraj, M. Sangareswari, V. Sailaja, Kapil Sihag

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The present study aimed to estimate the carbon sequestration potential of major forest types present in the foothills of Nilgiri biosphere reserve. The total biomass carbon stock was estimated in tropical thorn forest, tropical dry deciduous forest and tropical moist deciduous forest as 14.61 t C ha⁻¹ 75.16 t C ha⁻¹ and 187.52 t C ha⁻¹ respectively. The density and basal area were estimated in tropical thorn forest, tropical dry deciduous forest, tropical moist deciduous forest as 173 stems ha⁻¹, 349 stems ha⁻¹, 391 stems ha⁻¹ and 6.21 m² ha⁻¹, 31.09 m² ha⁻¹, 67.34 m² ha⁻¹ respectively. The soil carbon stock of different forest ecosystems was estimated, and the results revealed that tropical moist deciduous forest (71.74 t C ha⁻¹) accounted for more soil carbon stock when compared to tropical dry deciduous forest (31.80 t C ha⁻¹) and tropical thorn forest (3.99 t C ha⁻¹). The tropical moist deciduous forest has the maximum annual leaf litter which was 12.77 t ha⁻¹ year⁻¹ followed by 6.44 t ha⁻¹ year⁻¹ litter fall of tropical dry deciduous forest. The tropical thorn forest accounted for 3.42 t ha⁻¹ yr⁻¹ leaf litter production. The leaf litter carbon stock of tropical thorn forest, tropical dry deciduous forest and tropical moist deciduous forest found to be 1.02 t C ha⁻¹ yr⁻¹ 2.28 t⁻¹ C ha⁻¹ yr⁻¹ and 5.42 t C ha⁻¹ yr⁻¹ respectively. The results explained that decomposition percent at the soil surface in the following order.tropical dry deciduous forest (77.66 percent) > tropical thorn forest (69.49 percent) > tropical moist deciduous forest (63.17 percent). Decomposition percent at soil subsurface was studied, and the highest decomposition percent was observed in tropical dry deciduous forest (80.52 percent) followed by tropical moist deciduous forest (77.65 percent) and tropical thorn forest (72.10 percent). The decomposition percent was higher at soil subsurface. Among the three forest type, tropical moist deciduous forest accounted for the highest bacterial (59.67 x 105cfu’s g⁻¹ soil), actinomycetes (74.87 x 104cfu’s g⁻¹ soil) and fungal (112.60 x10³cfu’s g⁻¹ soil) population. The overall observation of the study helps to conclude that, the tropical moist deciduous forest has the potential of storing higher carbon content as biomass with the value of 264.68 t C ha⁻¹ and microbial populations.

Keywords: basal area, carbon sequestration, carbon stock, Nilgiri biosphere reserve

Procedia PDF Downloads 132