Search results for: AM Fungi

Authors: V. Shapaval, N. K. Afseth, D. Tzimorotas, A. Kohler

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Globally there is a dramatic increase in the demand for food, energy, materials and clean water since natural resources are limited. As a result, industries are looking for ways to reduce rest materials and to improve resource efficiency. Microorganisms have a high potential to be used as bio factories for the production of primary and secondary metabolites that represent high-value bio-products (enzymes, polyunsaturated fatty acids, bio-plastics, glucans, etc.). In order to find good microbial producers, to design suitable substrates from food rest materials and to optimize fermentation conditions, rapid analytical techniques for quantifying target bio products in microbial cells are needed. In the EU project FUST (R4SME, Fp7), we have developed a fully automated high-throughput FUST system based on micro-cultivation and FTIR spectroscopy that facilitates the screening of microorganisms, substrates and fermentation conditions for the optimization of the production of different high-value metabolites (single cell oils, bio plastics). The automated system allows the preparation of 100 samples per hour. Currently, The FUST system is in use for screening of filamentous fungi in order to find oleaginous strains with the ability to produce polyunsaturated fatty acids, and the optimization of cheap substrates, derived from food rest materials, and the optimization of fermentation conditions for the high yield of single cell oil.

Keywords: FTIR spectroscopy, FUST system, screening, biotechnology

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Authors: O. A. Babalola, A. F Adekunle, F. Oladeji, A. T. Osungbade, O. A. Akinlaja

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Optimizing microbiological activities in an organic crop production system is crucial to the realization of optimum growth and development of the crops. Field and pot experiments were conducted to assess soil microbial activities, growth and yield of tomato varieties in response to 4 rates of composted plant and animal residues. The compost rates were 0, 5, 10 and 20 t ha-1, and improved Ibadan and Ibadan local constituted the varieties. Fungi population, microbial biomass nitrogen, cellulase and proteinase activities were significantly higher (P≤ 0.05) at the rhizosphere of the local variety than that of improved variety. This led to a significantly higher number of branches, plant height, leaf area, number of fruits and less days to maturity in the local variety. Furthermore, compost-amended soil had significantly higher microbial populations, microbial biomass N, P and C, enzyme activities, soil N, P and organic carbon than control, but amendment of 20 t ha-1 gave significantly higher values than other compost rates. Consequently, growth parameters and tissue N significantly increased in all compost treatments while dry matter yield and weight of fruits were significantly higher in soil amended with 20 t ha-1. Correlation analysis showed that microbial activities at 6 weeks after transplanting (6 WAT) were more consistently and highly correlated with growth and yield parameters. It was concluded that microbial activities could be optimized to improve the yield of the two tomato varieties in an organic production system, through the application of compost, particularly at 20 t ha-1.

Keywords: compost, microbial activities, microbial contribution, tomato growth and yield

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Authors: Ubani Onyedikachi, Atagana Harrison Ifeanyichukwu, Thantsha Mapitsi Silvester

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Components of oil sludge (PAHs) are known cytotoxic, mutagenic and potentially carcinogenic compounds also bacteria and fungi have been found to degrade PAHs to innocuous compounds. This study is aimed at measuring the effect of pig, cow, horse and poultry manures on the degradation in selected PAHs present in oil sludge. Soil spiked with oil sludge was co-composted differently with each manure in a ratio of 2:1 (w/w) spiked soil: manure and wood-chips in a ratio of 2:1 (w/v) spiked soil: wood-chips. Control was set up similar as the one above but without manure. The mixtures were incubated for 10 months at room temperature. Compost piles were turned weekly and moisture level was maintained at between 50% and 70%. Moisture level, pH, temperature, CO2 evolution and oxygen consumption were measured monthly and the ash content at the end of experimentation. Highest temperature reached was 27.5 °C in all compost heaps, pH ranged from 5.5 to 7.8 and CO2 evolution was highest in poultry manure at 18.78μg/dwt/day. Microbial growth and activities were enhanced; bacteria identified were Bacillus, Arthrobacter and Staphylococcus species. Percentage reduction in PAHs was measured using automated soxhlet extractor with Dichloromethane coupled with gas chromatography/mass spectrometry (GC/MS). Results from PAH measurements showed reduction between 77% and 99%. Co-composting of spiked soils with animal manures enhanced the reduction in PAHs.

Keywords: animal manures, bioremediation, co-composting, oil refinery sludge, PAHs

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Authors: R. Sarada Jayalakshmi Devi B. Bhaskar, S. Khayum Ahammed, T. N. V. K. V. Prasad

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Use of bioagents and biofungicides is safe to manage the plant diseases and to avoid human health hazards which improves food security. Myconanotechnology is the study of nanoparticles synthesis using fungi and their applications. The present work reports on preparation, characterization and antifungal activity of biogenic silver nanoparticles produced by the fungus Trichoderma sp. which was collected from groundnut rhizosphere. The culture filtrate of Trichoderma sp. was used for the reduction of silver ions (Ag+) in AgNO3 solution to the silver (Ag0) nanoparticles. The different ages (4 days, 6 days, 8 days, 12 days, and 15 days) of culture filtrates were screened for the synthesis of silver nanoparticles. Synthesized silver nanoparticles were characterized using UV-Vis spectrophotometer, particle size and zeta potential analyzer, Fourier Transform Infrared Spectrophotometer (FTIR) and Transmission Electron Microscopy. Among all the treatments the silver nitrate solution treated with six days aged culture filtrate of Trichoderma sp. showed the UV absorption peak at 440 nm with maximum intensity (0.59) after 24 hrs incubation. The TEM micrographs showed the spherical shaped silver nanoparticles with an average size of 30 nm. The antifungal activity of silver nanoparticles against Aspergillus niger causing collar rot disease in groundnut and aspergillosis in humans showed the highest per cent inhibition at 100 ppm concentration (74.8%). The results points to the usage of these mycogenic AgNPs in agriculture to control plant diseases.

Keywords: groundnut rhizosphere, Trichoderma sp., silver nanoparticles synthesis, antifungal activity

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Authors: Gabriel S. De Oliveira, Patricia P. Adriani, Christophe Moriseau, Bruce D. Hammock, Felipe S. Chambergo

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Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have high biotechnological interest for the drug design and chemistry transformation for industries. In this study, we describe the identification of substrates and inhibitors of epoxide hydrolase enzyme from the filamentous fungus Trichoderma reesei (TrEH), and these inhibitors showed the fungal growth inhibitory activity. We have used the cloned enzyme and expressed in E. coli to develop the screening in the library of fluorescent substrates with the objective of finding the best substrate to be used in the identification of good inhibitors for the enzyme TrEH. The substrate (3-phenyloxiranyl)-acetic acid cyano-(6-methoxy-naphthalen-2-yl)-methyl ester showed the highest specific activity and was chosen for the next steps of the study. The inhibitors screening was performed in the library with more than three thousand molecules and we could identify the 6 best inhibitors. The IC50 of these molecules were determined in nM and all the best inhibitors have urea or amide in their structure, because It has been recognized that these groups fit well in the hydrolase catalytic pocket of the epoxide hydrolases. Then the growth of T. reesei in PDA medium containing these TrEH inhibitors was tested, and fungal growth inhibition activity was demonstrated with more than 60% of inhibition of fungus growth in the assay with the TrEH inhibitor with the lowest IC50. Understanding how this EH enzyme from T. reesei responds to inhibitors may contribute for the study of fungal metabolism and drug design against pathogenic fungi.

Keywords: epoxide hydrolases, fungal growth inhibition, inhibitor, Trichoderma reesei

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Authors: Sadia Roshan, Kulsoom Sughra, Shazia Shamas, Shamaila Irum, Haleema Sadia

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To evaluate synergistic medicinal effects of Withania somnifera (Ashwaghandha) and Murraya koenigii (Curry pata) in vitro. Antimicrobial activity was determined using the disc diffusion method against five bacterial and two fungal strains. The antioxidant activity was evaluated by the DPPH assay. The antidiabetic activity was evaluated by alpha-glucosidase inhibition assay and alpha-amylase inhibition assay. Synergistic antibacterial activity was observed against all the strains of bacteria, either Gram-positive or Gram-negative and fungi under study conditions. The maximum antibacterial activity was displayed by combined extract against E. coli i.e. 26±0.4mm. Maximum antifungal activity was shown by combined extract against Aspergillus niger, i.e., 17.3±0.5mm. The antioxidant activity of the combined extract was also significant. Alpha-glucosidase inhibition and alpha-amylase inhibition assays also showed synergism. Results indicate that Withania somnifera and Murraya koengii have medicinal properties. The combined extract of both plants is more potent than their individual extracts, suggesting that these can work in synergism. The research suggests that different plant extracts could be used in combination to increase their medicinal activities by many folds, thus giving an insight into future use of herbal medication.

Keywords: withania somnifera, murraya koenigii, antimicrobial activity, gram-positive bacetria, gram-negative bacteria

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Authors: Munir Ashraf, Shagufta Riaz

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Personal protective equipment (PPE) and bioactive textiles are highly important for the health care of front line hospital workers, patients, and the general population to be safe from highly infectious diseases. This was even more critical in the wake of COVID-19 outbreak. Most of the medical textiles are inactive against various viruses and bacteria, hence there is a need to wash them frequently to avoid the spread of microorganisms. According to survey conducted by the world health organization, more than 500 million people get infected from hospitals, and more than 13 million died due to these hospitals’ acquired deadly diseases. The market available PPE are though effective against the penetration of pathogens and to kill bacteria but, they are not breathable and active against different viruses. Therefore, there was a great need to develop textiles that are not only effective against bacteria, fungi, and viruses but also are comfortable to the medical personnel and patients. In the present study, waterproof breathable, and biologically active textiles were developed using antiviral and antibacterial nanomaterials. These nanomaterials like TiO₂, ZnO, Cu, and Ag were immobilized at the surface of cotton fabric by using different silane coupling agents and electroless deposition that they retained their functionality even after 30 industrial laundering cycles. Afterwards, the treated fabrics were coated with a waterproof breathable film to prevent the permeation of liquid droplets, any particle or microorganisms greater than 80 nm. The developed cotton fabric was highly active against bacteria and viruses. The good durability of nanomaterials at the cotton surface after several industrial washing cycles makes this fabric an ideal candidate for bioactive textiles used in the medical field.

Keywords: antibacterial, antiviral, cotton, durable

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Authors: Victor Iorungwa Gwa, Ebenezer Jonathan Ekefan

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Potential of Trichoderma harzianum for biological control of postharvest fungal rot of white yam (Dioscorea rotundata Poir) tubers in storage was studied. Pathogenicity test revealed the susceptibility of healthy looking yam tubers to Aspergillus niger, Botryodiplodia theobromae, and Fusarium oxysporum f. sp. melonganae after fourteen days of inoculation. Treatments comprising A. niger, B. theobromae, and F. oxysporum each paired with T. harzianum and were arranged in completely randomized design and stored for five months. Experiments were conducted between December 2015 and April 2016 and December 2016 and April 2017. Results showed that tubers treated with the pathogenic fungi alone caused mean percentage rot of between 6.67 % (F. oxysporum) and 22.22 % (A. niger) while the paired treatments produced only between 2.22 % (T. harzianum by F. oxysporum) and 6.67 % (T. harzianum by A. niger). In the second year of storage, mean percentage rot was found to be between 13.33 % (F. oxysporum) and 28.89 % (A. niger) while in the paired treatment rot was only between 6.67 % (F. oxysporum) and 8.89% (A. niger). Tubers treated with antagonist alone produced 0.00 % and 2.22 % in the first and second year, respectively. Result revealed that there was a significant difference (P ≤ 0.05) in mean percentage rot between the first year and the second year except where B. theobromae was inoculated alone, A. niger and T. harzianum paired and B. theobromae and T. harzianum paired. The most antagonised fungus in paired treatment for both years was F. oxysporum f. sp. melonganae, while the least antagonised, was A. niger and B. theobromae. It is, therefore, concluded that T. harzianum has potentials to control rot causing pathogens of yam tubers in storage. This can compliment or provide better alternative ways of reducing rot in yam tubers than by the use of chemical fungicides which are not environmentally friendly.

Keywords: biological control, fungal rot, postharvest, Trichoderma harzianum, white yam

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Authors: Akida Mulyaningtyas, Wahyudi Budi Sediawan

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One decisive stage in bioethanol production from plant biomass is the hydrolysis of lignocellulosic materials into simple sugars such as glucose. The produced glucose is then fermented into ethanol. This stage is popularly done in biological method by using cellulase that is produced by certain fungi. As it is known, glucose is the main source of nutrition for most microorganisms. Therefore, cutting cellulose into glucose is actually an attempt of microorganism to provide nutrition for itself. So far, this phenomenon has received less attention while it is necessary to identify the quantity of sugar consumed by the microorganism. In this study, we examined the phenomenon of sugar consumption by microorganism on lignocellulosic hydrolysis. We used oil palm empty fruit bunch (OPEFB) as the source of lignocellulose and Aspergillus niger as cellulase-producing fungus. In Indonesia, OPEFB is plantation waste that is difficult to decompose in nature and causes environmental problems. First, OPEFB was pretreated with 1% of NaOH at 170 oC to destroy lignin that hindered A.niger from accessing cellulose. The hydrolysis was performed by growing A.niger on pretreated OPEFB in solid state to minimize the possibility of contamination. The produced glucose was measured every 24 hours for 9 days. We analyzed the kinetics of both reactions, i.e., hydrolysis and glucose consumption, simultaneously. The constants for both reactions were assumed to follow the Monod equation. The results showed that the reaction constant of glucose consumption (μC) was higher than of cellulose hydrolysis (μH), i.e., 11.8 g/L and 0.62 g/L for glucose consumption and hydrolysis respectively. However, in general, the reaction rate of hydrolysis is greater than of glucose consumption since the cellulose concentration as substrate in hydrolysis is much higher than glucose as substrate in the consumption reaction.

Keywords: Aspergillus niger, bioethanol, hydrolysis, kinetics

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Authors: Wendu Admasu, Assefa Sintayehu, Alemu Gezahgne, Zewdu Terefework

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Eucalyptus is the most widely planted forest tree species in the world. In Ethiopia, pathogenic fungi pose an increasing threat to Eucalyptus species. Due to limited research, there is insufficient information on the associated diseases and pathogens. This study investigated Eucalyptus diseases, the extent of their damage, and the causal fungal pathogens. A Eucalyptus disease survey was conducted in the Eucalyptus forestry areas of Ethiopia during the growth years 2019/20 and 2020/21. Disease assessment and sampling were carried out in eighteen plantations at nine locations. E. camaldulensis was the most dominant species planted in the surveyed areas. The field study shows a high incidence and severity of canker diseases. Diseased stem and branch samples were collected, cultured on malt extract agar media and studied. The results of morphological and ITS sequence analysis confirmed that the fungal species Neofusicoccum parvum, Lasiodiplodia theobromae, and Aplosporella hesperidica caused the observed canker symptoms. This is the first report of Lasiodiplodia theobromae and Aplosporella hesperidica causing diseases in Eucalyptus plants in Ethiopia. Changes in global climate and environmental factors, such as altitude, are believed to have a strong impact on the susceptibility of Eucalyptus plants to diseases. Strict quarantine practices and continuous monitoring of pathogenic and endophytic fungal species associated with Eucalyptus trees are issued to be prioritized to effectively control and manage the disease.

Keywords: Neofusicoccum, Lasiodiplodia, Aplosporella, pathogenicity, phylogeny, severity

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Authors: Howaida I. Abd-Alla, Nagwa M. M. Shalaby

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The fruit rind of Fortunella margarita (Nagami Kumkuat) was investigated for its chemical constituents. Thirteen metabolites were obtained and classified into, a sterol; β-sitosterol (1) and twelve phenolic compounds, three coumarins; xanthotoxin (2), isopimpinellin (3), umbelliferone (4), nine flavonoids of O-glycosides of flavone; apigenin-7-O-β-D-glucopyranoside (5), apigenin-7-O-rhamnoglucoside (rhoifolin) (6), C-glycosides; vitexin (7), vicenin II (8), and the methoxylated; 6-methoxyapigenin-7-methyl ether (9) and tangeretin (10) as well as flavanones class; naringenin (11), liquiritigenin (12), hesperdin (hesperetin-7-rhamnoglucoside) (13). All compounds were identified for the first time in F. margarita except compound (8). The major glycosides 5, 6, and 13 and total crude extract showed potential antiviral activity against live Newcastle disease virus vaccine strains (Komarov and LaSota) and live infectious bursitis viruses vaccine strain D78 replication in VERO cell cultures and on specific pathogen-free embryonated chicken eggs. Antiviral inhibitory concentration fifty (IC50), cytotoxic concentration fifty (CC50), and therapeutic index (TI) were calculated. In addition, the extract and compounds 7 and 13 showed marked antimicrobial activity against different strains of fungi, Gram-positive and negative bacteria, including some foodborne pathogens of animal origin, caused human disease. These results suggested that the extract of F. margarita may be considered potentially useful as a source of natural antiviral and antimicrobial agents. It can be used as an ingredient for functional food and/or pharmaceuticals.

Keywords: antimicrobial, antiviral, Fortunella margarita, Nagami Kumkuat, phenolic secondary metabolites

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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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Authors: Tereza Branyšová, Martina Kračmarová, Kateřina Demnerová, Michal Ďurovič, Hana Stiborová

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Microbial deterioration threatens all objects of cultural heritage, including audio-visual materials. Fungi are commonly known to be the main factor in audio-visual material deterioration. However, although being neglected, bacteria also play a significant role. In addition to microbial contamination of materials, it is also essential to analyse air as a possible contamination source. This work aims to identify bacterial species in the archives of the Czech Republic that occur on audio-visual materials as well as in the air in the archives. For sampling purposes, the smears from the materials were taken by sterile polyurethane sponges, and the air was collected using a MAS-100 aeroscope. Metagenomic DNA from all collected samples was immediately isolated and stored at -20 °C. DNA library for the 16S rRNA gene was prepared using two-step PCR and specific primers and the concentration step was included due to meagre yields of the DNA. After that, the samples were sent to the University of Fairbanks, Alaska, for Illumina MiSeq sequencing. Subsequently, the analysis of the sequences was conducted in R software. The obtained sequences were assigned to the corresponding bacterial species using the DADA2 package. The impact of air contamination and the impact of different photosensitive layers that audio-visual materials were made of, such as gelatine, albumen, and collodion, were evaluated. As a next step, we will take a deeper focus on air contamination. We will select an appropriate culture-dependent approach along with a culture-independent approach to observe a metabolically active species in the air. Acknowledgment: This project is supported by grant no. DG18P02OVV062 of the Ministry of Culture of the Czech Republic.

Keywords: cultural heritage, Illumina MiSeq, metagenomics, microbial identification

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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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Authors: Kyung-Tae Lee, Li Li Wang, Kwang-Woo Jung, Yong-Sun Bahn

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Microtubules are involved in mechanical support, cytoplasmic organization as well as in a number of cellular processes by interacting with diverse microtubule-associated proteins (MAPs), such as plus-end tracking proteins, motor proteins, and tubulin-folding cofactors. A common feature of these proteins is the presence of a cytoskeleton-associated protein-glycine-rich (CAP-Gly) domain, which is evolutionarily conserved and generally considered to bind to α-tubulin to regulate functions of microtubules. However, there has been a dearth of research on CAP-Gly proteins in fungal pathogens, including Cryptococcus neoformans, which causes fatal meningoencephalitis globally. In this study, we identified five CAP-Gly proteins encoding genes in C. neoformans. Among these, Cgp1, encoded by CNAG_06352, has a unique domain structure that has not been reported before in other eukaryotes. Supporting the role of Cpg1 in microtubule-related functions, we demonstrate that deletion or overexpression of CGP1 alters cellular susceptibility to thiabendazole, a microtubule destabilizer, and Cgp1 is co-localized with cytoplasmic microtubules. Related to the cellular functions of microtubules, Cgp1 also governs maintenance of membrane stability and genotoxic stress responses. Furthermore, we demonstrate that Cgp1 uniquely regulates sexual differentiation of C. neoformans with distinct roles in the early and late stage of mating. Our domain analysis reveals that the CAP-Gly domain plays major roles in all the functions of Cgp1. Finally, the cgp1Δ mutant is attenuated in virulence. In conclusion, this novel CAP-Gly protein, Cgp1, has pleotropic roles in regulating growth, stress responses, differentiation and pathogenicity of C. neoformans.

Keywords: human fungal pathogen, CAP-Glycine protein, microtubule, meningoencephalitis

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

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Lignin, a heterogeneous three-dimensional biopolymer, is one of the building blocks of lignocellulosic biomass. Due to its limited chemical reactivity, lignin is currently processed as a low-value by-product in pulp and paper mills. Among various industrial lignins, Kraft lignin represents a major source of by-products generated during the widely employed pulping process across the pulp and paper industry. Therefore, valorization of Kraft lignin holds great potential as this would provide a readily available source of aromatic compounds for various industrial applications. Microbial degradation is well known for using both highly specific ligninolytic enzymes secreted by microorganisms and mild operating conditions compared with conventional chemical approaches. In this study, the degradation of Indulin AT lignin was assessed by comparing the effects of Basidiomycetous fungi (Coriolus versicolour and Trametes gallica) and Actinobacteria (Mycobacterium sp. and Streptomyces sp.) to two commercial laccases, T. versicolour ( ≥ 10 U/mg) and C. versicolour ( ≥ 0.3 U/mg). After 54 days of cultivation, the extent of microbial degradation was significantly higher than that of commercial laccases, reaching a maximum of 38 wt% degradation for C. versicolour treated samples. Lignin degradation was further confirmed by thermal carbon analysis with a five-step temperature protocol. Compared with commercial laccases, a significant decrease in char formation at 850ºC was observed among all microbial-degraded lignins with a corresponding carbon percentage increase from 200ºC to 500ºC. To complement the carbon analysis result, chemical characterization of the degraded products at different stages of the delignification by microorganisms and commercial laccases was performed by Pyrolysis-GC-MS.

Keywords: lignin, microbial degradation, pyrolysis-GC-MS, thermal carbon analysis

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Authors: Hanane Boutaj, Abdelilah Meddich, Said Wahbi, Zainab El Alaoui-Talibi, Allal Douira, Abdelkarim Filali-Maltouf, Cherkaoui El Modafar

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The present work aims to investigate the effect of arbuscular mycorrhizal fungi (AMF) in improving the olive tree resistance to Verticillium wilt caused by Verticillium dahliae. Inoculated plants with a mycorrhizal autochthonous consortium 'Rhizolive consortium' and pure strain 'Glomus irregulare' were infected after three months with V. dahliae. The improving of olive tree resistance was determined through disease severity, incidence, and defoliation. On the other hand, the defense mechanisms of olive plants were evaluated through lignin content, phenylalanine ammonia lyase (PAL) activity, and polyphenol content. The results revealed that both AMF significantly (p < 0.05) reduced disease development and the rate of defoliation in infected olive plants. Moreover, the contents of lignin were boosted after mycorrhizal inoculation in both the roots and the stems of olive plants, which remained significantly (p < 0.001) higher after the 90th days of V. dahliae inoculation. PAL activity was increased after V. dahliae inoculation in the stems of 'Rhizolive consortium' treatment that were 17 times higher than those in the roots of olive plants. The polyphenol content in the stems was about twice higher than those in the roots. The reduction of disease severity was accompanied by increased levels of lignin content, PAL activity, and polyphenol content, particularly in the stems of olive plants, indicating the strengthening of the olive plant immune system against V. dahliae.

Keywords: olive tree, Mycorrhizal autochthonous consortium, Glomus irregulare, Verticillium dahliae, defense mechanisms

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Authors: Uditi Dhakad, Baldev Ram, Chaman K. Jadon, R. K. Yadav, D. L. Yadav, Pratap Singh, Shalini Meena

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A field experiment was conducted during Kharif 2021 at Agricultural Research Station, Kota, to evaluate the effect of different post-emergence herbicides applied to soybean [Glycine max (L.) Merrill] on soil enzymes activity viz. dehydrogenase, phosphatase, and urease. The soil of the experimental site was clay loam (vertisols) in texture and slightly alkaline in reaction with 7.7 pH. The soil was low in organic carbon (0.49%), medium in available nitrogen (210 kg/ha), phosphorus (23.5 P2O5 kg/ha), and high in potassium (400 K2O kg/ha) status. The results elucidated that no significant adverse effect on soil dehydrogenase, urease, and phosphatase activity was determined with the application of post-emergence herbicides over the untreated control. Two hands weeding at 20 and 40 DAS registered maximum dehydrogenase enzyme activity (0.329 μgTPF/g soil/d) closely followed by herbicides mixtures and sole herbicide while pre-emergence application of pendimethalin + imazethapyr 960 g a.i./ha and pendimethalin 1.0 kg a.i./ha significantly reduced dehydrogenase enzyme activity compared to control. Urease enzyme activity was not much affected under different weed control treatments and weedy checks. The treatments were found statistically non-significant, and values ranged between 1.16-1.25 μgNH4N/g soil/d. Phosphatase enzyme activity was also not influenced significantly due to various weed control treatments. Though maximum phosphatase enzyme activity (30.17 μgpnp/g soil/hr) was observed under two-hand weeding, followed by fomesafen + fluazifop-p-butyl 220 g a.i./ha. Herbicidal weed control measures did not influence the total bacteria, fungi, and actinomycetes population.

Keywords: dehydrogenase, phosphatase, post-emergence, soil enzymes, urease.

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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

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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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Authors: Ikpesu, Thomas Ohwofasa, Babtunde Ilesanmi

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The effects of anthropogenic activities on fish and fisheries products in Niger Delta water bodies were investigated. The rivers were selected based on their close proximity to contaminants and dredging activities. Three stations were chosen per river. The stations chosen to depicting downstream and upstream stations were visited and samples collected on monthly basis. The down streams stations are the polluted and heavily dredged sites, where the upstream station is far, without any evidence of pollution or human activities. During these periods, the fishes of the same species were collected and analyzed for morphological and physiological changes, after which they were returned back to the rivers. The physico-chemicals parameters of these stations were also taken. Morphological changes such as skin ulcerations and other lesions, as well as fungi infections were observed in the down streams fishes. The fish in up streams look healthier and bigger (though the age could not be affirmed) than the downstream fishes. The physico-chemical parameters between the up streams and down streams stations vary significantly (p < 0.01). These anthropogenic effects must have interfere with the normal migration pattern of these fishes, because there were changes in the composition of population and species diversity in the samples sites, with the upstream having true species diversity. The release of pollutants into the water in the Niger Delta areas may triggers off naturally occurring bio toxicity cycles and other fish poisoning. There is risk of biomagnifications of these poisons along the tropic level. This makes the normally valuable food resource dangerous for human consumption and thereby instances of human death caused by such poisoning.

Keywords: anthropogenic, dredging, fisheries, niger delta, pollution, rivers

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Authors: Q. Rzina, M. Lahrouni, S. Rida, N. Saadaoui, Y. Almossaid, K. Oufdou, K. Fares

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Many organic solid wastes are produced in the world. Poultry manure (PM), municipal organic wastes (MOW) and sugar beet lime sludge (LS) are produced in large quantities in Morocco. The co-composting of these organic wastes was investigated. The recycling and the valorization of such wastes is environmentally and economically beneficial especially for PM which is known source of bacterial pathogens. The aerobic biodegradation process was carried out by using three windrows of variable compositions: C1 prepared without LS (only MOW were composted with PM), C2 prepared from MOW plus PM and10% LS; and the last one C3 from MOW plus PM and 20% LS. The main process physico-chemical parameters (temperature, pH, humidity and C/N) and microbiological populations (mesophilic and thermophilic flora, total coliform, fecal coliform, Streptococci, Staphylococcus aureus and mesophilic fungi) were monitored over three months to ascertain the compost maturity and to ensure the compost hygienic aspect. The final products were characterized by their relatively high organic matter content, and low C/N ratio of 10.6-10.9. The organic matter degradation was reached approximately 59% for C2 and C3. In addition, the monitoring of the microbial population showed that the produced composts are mature and hygienic. The agronomic valorization of the final composts was tested on radish plant with tree level of composts and poultry manure without composting. The primary results of field trial showed a growth of radish plant biomass and root development without any phytotoxicity detected which reflects the quality of the composts produced. As for poultry manure it allowed to have a better results than other composts because of its readily available nitrogen.

Keywords: compost, municipal organic wastes, poultry manure, radish crop, sugar beet lime sludge

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Authors: Muhammad Adeel Arshad, Faiz-Ul Hassan, Yanfen Cheng

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According to FAO, enteric methane (CH4) production is about 44% of all greenhouse gas emissions from the livestock sector. Ruminants produce CH4 as a result of fermentation of feed in the rumen especially from roughages which yield more CH4 per unit of biomass ingested as compared to concentrates. Efficient ruminal fermentation is not possible without abating CO2 and CH4. Methane abatement strategies are required to curb the predicted rise in emissions associated with greater ruminant production in future to meet ever increasing animal protein requirements. Ecology of ruminal methanogenesis and avenues for its mitigation can be identified through various genomic and transcriptomic techniques. Programs such as Hungate1000 and the Global Rumen Census have been launched to enhance our understanding about global ruminal microbial communities. Through Hungate1000 project, a comprehensive reference set of rumen microbial genome sequences has been developed from cultivated rumen bacteria and methanogenic archaea along with representative rumen anaerobic fungi and ciliate protozoa cultures. But still many species of rumen microbes are underrepresented especially uncultivable microbes. Lack of sequence information specific to the rumen's microbial community has inhibited efforts to use genomic data to identify specific set of species and their target genes involved in methanogenesis. Metagenomic and metatranscriptomic study of entire microbial rumen populations offer new perspectives to understand interaction of methanogens with other rumen microbes and their potential association with total gas and methane production. Deep understanding of methanogenic pathway will help to devise potentially effective strategies to abate methane production while increasing feed efficiency in ruminants.

Keywords: Genome sequences, Hungate1000, methanogens, ruminal fermentation

Procedia PDF Downloads 139

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

Procedia PDF Downloads 153

Authors: Asmuddin Natsir, A. Mujnisa, Syahriani Syahrir, Marhamah Nadir, Nurul Purnomo

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Bacteria, protozoa, Archaea, and fungi are the dominant microorganisms found in the rumen ecosystem that has an important role in converting feed ingredients into components that can be digested and utilized by the livestock host. This study was conducted to assess the diversity of rumen bacteria of bali cattle raised under traditional farming condition. Three adult bali cattle were used in this experiment. The rumen fluid samples from the three experimental animals were obtained by the Stomach Tube method before the morning feeding. The results of study indicated that the Illumina sequencing was successful in identifying 301,589 sequences, averaging 100,533 sequences, from three rumen fluid samples of three cattle. Furthermore, based on the SILVA taxonomic database, there were 19 kinds of phyla that had been successfully identified. Of the 19 phyla, there were only two dominant groups across the three samples, namely Bacteroidetes and Firmicutes, with an average percentage of 83.68% and 13.43%, respectively. Other groups such as Synergistetes, Spirochaetae, Planctomycetes can also be identified but in relatively small percentage. At the genus level, there were 157 sequences obtained from all three samples. Of this number, the most dominant group was Prevotella 1 with a percentage of 71.82% followed by 6.94% of Christencenellaceae R-7 group. Other groups such as Prevotellaceae UCG-001, Ruminococcaceae NK4A214 group, Sphaerochaeta, Ruminococcus 2, Rikenellaceae RC9 gut group, Quinella were also identified but with very low percentages. The sequencing results were able to detect the presence of 3.06% and 3.92% respectively for uncultured rumen bacterium and uncultured bacterium. In conclusion, the results of this experiment can provide an opportunity for a better understanding of the rumen bacterial diversity of the bali cattle raised under traditional farming condition and insight regarding the uncultured rumen bacterium and uncultured bacterium that need to be further explored.

Keywords: 16S rRNA sequencing, bali cattle, rumen microbial diversity, uncultured rumen bacterium

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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

Procedia PDF Downloads 147

Authors: Dimphna Ezikanyi, Gloria Sakwari

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Airborne pollen and fungal spores are major triggers of allergies, and their abundance and seasonality depend on plant responses to climatic and meteorological variables. A survey of seasonal prevalence of airborne pollen and fungal spores in Nsukka, Enugu, South- East Nigeria and relationship to climatic variables were carried out from Jan-June, 2017. The aim of the study was to access climate change and variability over time in the area and their accrued influence on modern pollen and spores rain. Decadal change in climate was accessed from variables collected from meteorological centre in the study area. Airborne samples were collected monthly using a modified Tauber-like pollen samplers raised 5 ft above ground level. Aerosamples collected were subjected to acetolysis. Dominant pollen recorded were those of Poaceae, Elaeis guinensis Jacq. and Casuarina equisetifolia L. Change in weather brought by onset of rainfall evoked sporulation and dispersal of diverse spores into ambient air especially potent allergenic spores with the spores of Ovularia, Bispora, Curvularia, Nigrospora, Helminthosporium preponderant; these 'hydrophilic fungi' were abundant in the rainy season though in varying quantities. Total fungal spores correlated positively with monthly rainfall and humidity but negatively with temperature. There was a negative though not significant correlation between total pollen count and rainfall. The study revealed a strong influence of climatic variables on abundance and spatial distribution of pollen and fungal spores in the ambient atmosphere.

Keywords: allergy, fungal spores, pollen, weather parameters

Procedia PDF Downloads 176

Authors: Sameen Tariq, Saira Bano, Sayyada Ghufrana Nadeem

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The Ocean, which covers over 70% of the world's surface, is wealthy in biodiversity as well as a rich wellspring of microorganisms with huge potential. The oceanic climate is home to an expansive scope of plants, creatures, and microorganisms. Marine microbial networks, which incorporate microscopic organisms, infections, and different microorganisms, enjoy different benefits in biotechnological processes. Samples were collected from marine environments, including soil and water samples, to cultivate the uncultured marine organisms by using Zobell’s medium, Sabouraud’s dextrose agar, and casein media for this purpose. Following isolation, we conduct microscopy and biochemical tests, including gelatin, starch, glucose, casein, catalase, and carbohydrate hydrolysis for further identification. The results show that more gram-positive and gram-negative bacteria. The isolation process of marine organisms is essential for understanding their ecological roles, unraveling their biological secrets, and harnessing their potential for various applications. Marine organisms exhibit remarkable adaptations to thrive in the diverse and challenging marine environment, offering vast potential for scientific, medical, and industrial applications. The isolation process plays a crucial role in unlocking the secrets of marine organisms, understanding their biological functions, and harnessing their valuable properties. They offer a rich source of bioactive compounds with pharmaceutical potential, including antibiotics, anticancer agents, and novel therapeutics. This study is an attempt to explore the diversity and dynamics related to marine microflora and their role in biofilm formation.

Keywords: marine microorganisms, ecosystem, fungi, biofilm, gram-positive, gram-negative

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

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A variety of genetic and environmental factors cause various cosmetics and dermatological problems. There are already claimed drugs available in market for treating these problems. However, the challenge remains in finding more potent, environmental friendly, causing minimal side effects and economical cosmeceuticals. This leads to an increased demand for natural cosmeceutical products in the last few decades. Plant derived ingredients are limited because plants either contain toxic metabolites, grow too slow or seasonal harvesting is a problem. The research work carried out in this project aims at isolation, characterization of marine fungal secondary metabolite and evaluating their potential use in future cosmetic skin care products. We have isolated and purified 35 morphologically different fungal isolates from various marine habitats of the India. These isolates have been functionally characterized for anti-tyrosinase, antioxidant and anti-acne activities. For molecular characterization, the Internal Transcribed spacer (ITS) region of 15 functionally active marine fungal isolates was amplified using universal primers, ITS1 and ITS4 and sequenced. Out of 15 marine fungal isolates crude extract of strains D4 (Aspergillus terreus) and P2 (Talaromyces stipitatus) showed 70% and 57% tyrosinase inhibition at 1mg/mL respectively. Strain D5 (Simplicillium lamellicola) has showed significant inhibition against Propionibacterium acnes and Staphylococcus epidermidis. In addition, all these strains also displayed DPPH- radical scavenging activity and may be utilized as skin cosmeceutical applications. Purification and characterization of crude extracts for identification of active lead molecule is under process.

Keywords: anti-acne, anti-tyrosinase, cosmeceutical, marine fungi

Procedia PDF Downloads 277

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

Procedia PDF Downloads 62