Search results for: Aristotle on being is not a genus

Authors: Howaida I. Abd-Alla, Omnia Kutkat, Yassmin Moatasim, Magda T. Ibrahim, Marwa A. Mostafa, Mohamed GabAllah, Mounir M. El-Safty

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Artemisia judaica (known shih-balady), Azadirachta indica and Sophora tomentosa (known yellow necklace pod) are members of available medicinal plants well-known for their traditional medical use in Egypt which suggests that they probably harbor broad-spectrum antiviral, immunostimulatory and anti-inflammatory functions. Their ethyl acetate-dichloromethane (1:1, v/v) extracts were evaluated for the potential anti-Middle East respiratory syndrome-related coronavirus (anti-MERS-CoV) activity. Their cytotoxic activity was tested in Vero-E6 cells using 3-(4,-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method with minor modification. The plot of percentage cytotoxicity for each extract concentration has calculated the concentration which exhibited 50% cytotoxic concentration (TC50). A plaque reduction assay was employed using safe dose of extract to evaluate its effect on virus propagation. The highest inhibition percentage was recorded for the yellow necklace pod, followed by Shih-balady. The possible mode of action of virus inhibition was studied at three different levels viral replication, viral adsorption and virucidal activity. The necklace pod leaves have induced virucidal effects and direct effects on the replication of virus. Phytochemical investigation of the promising necklace pod led to the isolation and structure determination of nine compounds. The structure of each compound was determined by a variety of spectroscopic methods. Compounds 4-O-methyl sorbitol 1, 8-methoxy daidzin 6 and 6-methoxy apigenin-7-O-β-D-glucopyranoside 8 were isolated for the first time from the Sophora genus and the other six compounds were the first time that they were isolated from this species according to available works of literature. Generally, the highest anti-CoV 2 activity of S. tomentosa was associated with the crude ethanolic extract, indicating the possibility of synergy among the antiviral phytochemical constituents (1-9).

Keywords: coronavirus, MERS-CoV, mode of action, necklace pod, shih-balady

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Authors: Gerhardt Boukes, Maryna Van De Venter, Sharlene Govender

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Macrofungi have been used for the past two thousand years in Asian countries, and more recently in Western countries, for their medicinal properties. Biological activities include antimicrobial, antioxidant, anti-inflammatory, antidiabetic, anticancer and immunomodulatory to name a few. Several biologically active compounds have been identified and isolated. Macrofungal research in Africa is poorly documented and to the best of our knowledge non-existent. South Africa has a rich macrofungal biodiversity, which includes endemic and exotic macrofungal species. Ethanolic extracts of 13 macrofungal species, including mushrooms, bracket fungi and puffballs, were prepared and screened for cytotoxicity against a panel of seven cell lines, including A549 (human lung adenocarcinoma), HeLa (human cervical adenocarcinoma), HT-29 (human colorectal adenocarcinoma), MCF7 (human breast adenocarcinoma), MIA PaCa-2 (human pancreatic ductal adenocarcinoma), PC-3 (human prostate adenocarcinoma) and Vero (African green monkey kidney epithelial) cells using MTT. Cell lines were chosen according to the most prevalent cancer types affecting males and females in South Africa and globally, and the mutations they contain. Preliminary results have shown that three of the macrofungal genera, i.e. Fomitopsis, Gymnopilus and Pycnoporus, have shown cytotoxic activity, ranging between IC50 ~20 and 200 µg/mL. The molecular mechanism of action contributing to cell death investigated and being investigated include apoptosis (i.e. DNA cell cycle arrest, caspase-3 activation and mitochondrial membrane potential), autophagy (i.e. acridine orange and LC3B staining) and ER stress (i.e. thioflavin T staining and caspase-12) in the presence of melphalan, chloroquine and thapsigargin/tuncamycin as positive controls, respectively. The genus, Pycnoporus, has shown the best cytotoxicity of the three macrofungal genera. Future work will focus on the identification and isolation of novel active compounds and elucidating the mechanism/s of action.

Keywords: cancer, cytotoxicity, macrofungi, mechanism/s of action

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Authors: Lali Kutateladze, Tamar Urushadze, Tamar Dudauri, Besarion Metreveli, Nino Zakariashvili, Izolda Khokhashvili, Maya Jobava

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Lignocellulosic waste streams from agriculture, paper and wood industry are renewable, plentiful and low-cost raw materials that can be used for large-scale production of liquid and gaseous biofuels. As opposed to prevailing multi-stage biotechnological processes developed for bioconversion of cellulosic substrates to ethanol where high-cost cellulase preparations are used, Consolidated Bioprocessing (CBP) offers to accomplish cellulose and xylan hydrolysis followed by fermentation of both C6 and C5 sugars to ethanol in a single-stage process. Syntrophic microbial consortium comprising of anaerobic, thermophilic, cellulolytic, and saccharolytic bacteria in the genus Clostridia with improved ethanol productivity and high tolerance to fermentation end-products had been proposed for achieving CBP. 65 new strains of anaerobic thermophilic cellulolytic and saccharolytic Clostridia were isolated from different wetlands and hot springs in Georgia. Using new isolates, fermentation of mechanically pretreated wheat straw and corn stalks was done under oxygen-free nitrogen environment in thermophilic conditions (T=550C) and pH 7.1. Process duration was 120 hours. Liquid and gaseous products of fermentation were analyzed on a daily basis using Perkin-Elmer gas chromatographs with flame ionization and thermal detectors. Residual cellulose, xylan, xylose, and glucose were determined using standard methods. Cellulolytic and saccharolytic bacteria strains degraded mechanically pretreated herbaceous cellulosic wastes and fermented glucose and xylose to ethanol, acetic acid and gaseous products like hydrogen and CO2. Specifically, maximum yield of ethanol was reached at 96 h of fermentation and varied between 2.9 – 3.2 g/ 10 g of substrate. The content of acetic acid didn’t exceed 0.35 g/l. Other volatile fatty acids were detected in trace quantities.

Keywords: anaerobic bacteria, cellulosic wastes, Clostridia sp, ethanol

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Authors: Johanna Bapela, Heino Heyman, Marion Meyer

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Regardless of the significant advances accomplished in reducing the burden of malaria and other tropical diseases in recent years, malaria remains a major cause of mortality in endemic countries. This is especially the case in sub-Saharan Africa where 99% of the estimated global malaria deaths occurs on an annual basis. The emergence of resistant Plasmodium species and the lack of diversified chemotherapeutic agents provide the rationale for bioprospecting for antiplasmodial scaffolds. Crude extracts from twenty indigenous antimalarial plant species were screened for antimalarial activity and then subjected to 1H NMR-based metabolomic analysis. Ten plant extracts exhibited significant in vitro antiplasmodial activity (IC50 ≤ 5 µg/ml). The Principal Component Analysis (PCA) of the acquired 1H NMR spectra could not separate the analyzed plant extracts according to the detected antiplasmodial bioactivity. Application of supervised Orthogonal Projections to Latent Structures–Discriminant Analysis (OPLS-DA) to the 1H NMR profiles resulted in a discrimination pattern that could be correlated to bioactivity. A contribution plot generated from the OPLS-DA scoring plot illustrated the classes of compounds responsible for the observed grouping. Given the preliminary in vitro results, Tabernaemontana elegans Stapf. (Apocynaceae) and Vangueria infausta Burch. subsp. infausta (Rubiaceae) were subjected to further phytochemical investigations. Two indole alkaloids, dregamine and tabernaemontanine possessing antiplasmodial activity were isolated from T. elegans. Two compounds were isolated from V. infausta subsp. infausta and identified as friedelin (IC50 = 3.01 µg/ml) and morindolide (IC50 = 18.5 µg/ml). While these compounds have been previously identified, this is the first account of their occurrence in the genus Vangueria and their antiplasmodial activity. Based on the results of the study, metabolomics can be used to globally identify classes of plant secondary metabolites that are responsible for antiplasmodial activity.

Keywords: ethnopharmacology, Malaria, medicinal plants, metabolomics

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Authors: Chanya Jetsukontorn

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Dengue hemorrhagic fever is the most important Mosquito-borne disease and the major public health problem in Thailand. The most important vector is Aedes aegypti. Environmental factors such as temperature, relative humidity, and biting rate affect dengue virus infection. The most effective measure for prevention is controlling of vector mosquitoes. In addition, surveillance of field-caught mosquitoes is imperative for determining the natural vector and can provide an early warning sign at risk of transmission in an area. In this study, Aedes aegypti mosquitoes were collected in Amphur Muang, Phetchabun Province, Thailand. The mosquitoes were collected in the rainy season and the dry season both indoor and outdoor. During mosquito’s collection, the data of environmental factors such as temperature, humidity and breeding sites were observed and recorded. After identified to species, mosquitoes were pooled according to genus/species, and sampling location. Pools consisted of a maximum of 10 Aedes mosquitoes. 70 pools of 675 Aedes aegypti were screened with RT-PCR for flaviviruses. To confirm individual infection for determining True infection rate, individual mosquitoes which gave positive results of flavivirus detection were tested for dengue virus by RT-PCR. The infection rate was 5.93% (4 positive individuals from 675 mosquitoes). The probability to detect dengue virus in mosquitoes at the neighbour’s houses was 1.25 times, especially where distances between neighboring houses and patient’s houses were less than 50 meters. The relative humidity in dengue-infected villages with dengue-infected mosquitoes was significantly higher than villages that free from dengue-infected mosquitoes. Indoor biting rate of Aedes aegypti was 14.87 times higher than outdoor, and biting times of 09.00-10.00, 10.00-11.00, 11.00-12.00 yielded 1.77, 1.46, 0.68mosquitoes/man-hour, respectively. These findings confirm environmental factors were related to Dengue infection in Thailand. Data obtained from this study will be useful for the prevention and control of the diseases.

Keywords: Aedes aegypti, Dengue virus, environmental factors, one health, PCR

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Authors: Nurul Aliaa Idris, David A. Collings

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Phi thickenings are specialized secondary cell wall thickenings that are found in the cortex of the roots in a wide range of plant species, including orchids. The role of phi thickenings in the root is still under debate through research have linked environmental conditions, particularly abiotic stresses such as water stress, heavy metal stress and salinity to their induction in the roots. It has also been suggested that phi thickenings may act as a barrier to regulate solute uptake, act as a physical barrier against fungal hyphal penetration due to its resemblance to the Casparian strip and play a mechanical role to support cortical cells. We have investigated phi thickening function in epiphytic orchids of the genus Miltoniopsis through induction experiment against factors such as soil compaction and water stress. The permeability of the phi thickenings in Miltoniopsis was tested through uptake experiments using the fluorescent tracer dyes Calcofluor white, Lucifer yellow and Propidium iodide then viewed with wide-field or confocal microscopy. To test whether phi thickening may prevent fungal colonization in the root cell, fungal re-infection experiment was conducted by inoculating isolated symbiotic fungus to sterile in vitro Miltoniopsis explants. As the movement of fluorescent tracers through the apoplast was not blocked by phi thickenings, and as phi thickenings developed in the roots of sterile cultures in the absence of fungus and did not prevent fungal colonization of cortical cells, the phi thickenings in Miltoniopsis do not function as a barrier. Phi thickenings were found to be absent in roots grown on agar and remained absent when plants were transplanted to moist soil. However, phi thickenings were induced when plants were transplanted to well-drained media, and by the application of water stress in all soils tested. It is likely that phi thickenings stabilize the root cortex during dehydration. Nevertheless, the varied induction responses present in different plant species suggest that the phi thickenings may play several adaptive roles, instead of just one, depending on species.

Keywords: abiotic stress, Miltoniopsis, orchid, phi thickening

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Authors: Sergio Alejandro Cuevas, Catherine Etchebest, Fernando Luis Barroso Da Silva

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The flavivirus genus has several organisms responsible for generating various diseases in humans. Special in Brazil, Zika (ZIKV), Dengue (DENV) and Yellow Fever (YFV) viruses have raised great health concerns due to the high number of cases affecting the area during the last years. Diagnostic is still a difficult issue since the clinical symptoms are highly similar. The understanding of their common structural/dynamical and biomolecular interactions features and differences might suggest alternative strategies towards differential serological diagnostics and therapeutic intervention. Due to their immunogenicity, the primary focus of this study was on the ZIKV, DENV and YFV non-structural proteins 1 (NS1) protein. By means of computational studies, we calculated the main physical chemical properties of this protein from different strains that are directly responsible for the biomolecular interactions and, therefore, can be related to the differential infectivity of the strains. We also mapped the electrostatic differences at both the sequence and structural levels for the strains from Uganda to Brazil that could suggest possible molecular mechanisms for the increase of the virulence of ZIKV. It is interesting to note that despite the small changes in the protein sequence due to the high sequence identity among the studied strains, the electrostatic properties are strongly impacted by the pH which also impact on their biomolecular interactions with partners and, consequently, the molecular viral biology. African and Asian strains are distinguishable. Exploring the interfaces used by NS1 to self-associate in different oligomeric states, and to interact with membranes and the antibody, we could map the strategy used by the ZIKV during its evolutionary process. This indicates possible molecular mechanisms that can explain the different immunological response. By the comparison with the known antibody structure available for the West Nile virus, we demonstrated that the antibody would have difficulties to neutralize the NS1 from the Brazilian strain. The present study also opens up perspectives to computationally design high specificity antibodies.

Keywords: zika, biomolecular interactions, electrostatic interactions, molecular mechanisms

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Authors: Yuda Purwana Roswanjaya, Wouter Kohlen, Rene Geurts

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The Trema genus represents a group of fast-growing tropical tree species within the Cannabaceae. Interestingly, five species nested in this lineage -known as Parasponia- can establish rhizobium nitrogen-fixing root nodules, similar to those found in legumes. Parasponia and legumes use a conserved genetic network to control root nodule formation, among which are genes also essential for mycorrhizal symbiosis (the so-called common symbiotic pathway). However, Trema species lost several genes that function exclusively in nodulation, suggesting a loss-of the nodulation trait in Trema. Strikingly, in a Trema orientalis population found in Malaysian Borneo we identified a truncated SYMBIOSIS RECEPTOR KINASE (SYMRK) mutant allele lacking a large portion of the c-terminal kinase domain. In legumes this gene is essential for nodulation and mycorrhization. This raises the question whether Trema orientalis can still be mycorrhized. To answer this question, we established quantitative mycorrhization assay for Parasponia andersonii and Trema orientalis. Plants were grown in closed pots on half strength Hoagland medium containing 20 µM potassium phosphate in sterilized sand and inoculated with 125 spores of Rhizopagus irregularis (Agronutrion-DAOM197198). Mycorrhization efficiency was determined by analyzing the frequency of mycorrhiza (%F), the intensity of the mycorrhizal colonization (%M) and the arbuscule abundance (%A) in the root system. Trema orientalis RG33 can be mycorrhized, though with lower efficiency compared to Parasponia andersonii. From this we conclude that a functional SYMRK kinase domain is not essential for Trema orientalis mycorrhization. In ongoing experiments, we aim to investigate the role of SYMRK in Parasponia andersonii mycorrhization and nodulation. For this two Parasponia andersonii symrk CRISPR-Cas9 mutant alleles were created. One mimicking the TorSYMRKRG33 allele by deletion of exon 13-15, and a full Parasponia andersonii SYMRK knockout.

Keywords: endomycorrhization, Parasponia andersonii, symbiosis receptor kinase (SYMRK), Trema orientalis

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Authors: Muhammad Dildar Gogi, Muhammad Jalal Arif, Muhammad Junaid Nisar, Mubashir Iqbal, Waleed Afzal Naveed, Muhammad Ahsan Khan, Ahmad Nawaz, Muhammad Sufian, Muhammad Arshad, Amna Jalal

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Fruit flies of Bactrocera genus cause heavy losses in fruits and vegetables globally and insecticide-application for their control creates issues of ecological backlash, environmental pollution, and food safety. There is need to explore alternatives and food-baits application is considered safe for the environment and effective for fruit fly management. Present experiment was carried out to assess the attractancy of five phagostimulant-Mixtures (PHS-Mix) prepared by mixing banana-squash, mulberry, protein-hydrolysate and molasses with some phagostimulant-lure sources including beef extract, fish extract, yeast, starch, rose oil, casein and cedar oil in five different ratios i.e., PHS-Mix-1 (1 part of all ingredients), PHS-Mix-2 (1 part of banana with 0.75 parts of all other ingredients), PHS-Mix-3 (1 part of banana with 0.5 parts of all other ingredients), PHS-Mix-4 (1 part of banana with 0.25 parts of all other ingredients) and PHS-Mix-5 (1 part of banana with 0.125 parts of all other ingredients). These were evaluated in comparison with a standard (GF-120). PHS-Mix-4 demonstrated 40.5±1.3-46.2±1.6% AI for satiated flies (class-II i.e., moderately attractive) and 59.5±2.0-68.6±3.0% AI for starved flies (class-III i.e., highly attractive) for both B. dorsalis and B. zonata in olfactometric study while the same exhibited 51.2±0.53% AI (class-III i.e., highly attractive) for B. zonata and 45.4±0.89% AI (class-II i.e., moderately attractive) for B. dorsalis in field study. PHS-Mix-1 proved non-attractive (class-I) and moderately attractive (class-II) phagostimulant in olfactometer and field studies, respectively. PHS-Mix-2 exhibited moderate attractiveness for starved lots in olfactometer and field-lot in field studies. PHS-Mix-5 proved non-attractive to starved and satiated lots of B. zonata and B. dorsalis females in olfactometer and field studies. Overall PHS-Mix-4 proved better phagostimulant-mixture followed by PHS-Mix-3 which was categorized as class-II (moderately attractive) phagostimulant for starved and satiated lots of female flies of both species in olfactometer and field studies; hence these can be exploited for fruit fly management.

Keywords: attractive index, field conditions, olfactometer, Tephritid flies

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Authors: Hadiatullah, T. S. D. Desak Ketut, A. A. Ayu, A. N. Isna, D. P. Ririn

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Spirogyra sp. is genus of microalgae which has a high carbohydrate content that used as a best medium for bacterial fermentation to produce cellulose. This study objective to determine the effect of pulp banana in the fermented paper production process using Spirogyra sp. and characterizing of the paper product. The method includes the production of bacterial cellulose, assay of the effect fermented paper interpolating with banana pulp using Spirogyra sp., and the assay of paper characteristics include gram-mage paper, water assay absorption, thickness, power assay of tensile resistance, assay of tear resistance, density, and organoleptic assay. Experiments were carried out with completely randomized design with a variation of the concentration of sewage treatment in the fermented paper production interpolating banana pulp using Spirogyra sp. Each parameter data to be analyzed by Anova variance that continued by real difference test with an error rate of 5% using the SPSS. Nata production results indicate that different carbon sources (glucose and sugar) did not show any significant differences from cellulose parameters assay. Significantly different results only indicated for the control treatment. Although not significantly different from the addition of a carbon source, sugar showed higher potency to produce high cellulose. Based on characteristic assay of the fermented paper showed that the paper gram-mage indicated that the control treatment without interpolation of a carbon source and a banana pulp have better result than banana pulp interpolation. Results of control gram-mage is 260 gsm that show optimized by cardboard. While on paper gram-mage produced with the banana pulp interpolation is about 120-200 gsm that show optimized by magazine paper and art paper. Based on the density, weight, water absorption assays, and organoleptic assay of paper showing the highest results in the treatment of pulp banana interpolation with sugar source as carbon is 14.28 g/m2, 0.02 g and 0.041 g/cm2.minutes. The conclusion found that paper with nata material interpolating with sugar and banana pulp has the potential formulation to produce super-quality paper.

Keywords: cellulose, fermentation, grammage, paper, Spirogyra sp.

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Authors: H. Alijani, M. Jabari, S. Matroodi, H. Zolqarnein, A. Sharafi, I. Zamani

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Actinomycetes, Gram-positive bacteria, are interesting as a main producer of secondary metabolites and are important industrially and pharmaceutically. The marine environment is a potential source for new actinomycetes, which can provide novel bioactive compounds and industrially important enzymes. The aims of this study were to isolate and identify novel actinomycetes from Persian Gulf sediments and screen these isolates for the production of secondary metabolites, especially antibiotics, Using phylogenetic (16S rRNA gene sequence), morphological and biochemical analyses. 15 different actinomycete strains from Persian Gulf sediments at a depth of 5-10 m were identified. DNA extraction was done using Cinnapure DNA Kit. PCR amplification of 16S rDNA gene was performed using F27 and R1492 primers. Phylogenetic tree analysis was performed using the MEGA 6 software. Most of the isolated strains belong to the genus namely Streptomyces (14), followed by Nocardiopsis (1). Antibacterial assay of the isolates supernatant was performed using a standard disc diffusion assay with replication (n=3). The results of disk diffusion assay showed that most active strain against Proteus volgaris and Bacillus cereus was AMJ1 (16.46±0.2mm and 13.78±0.2mm, respectively), against Salmonella sp. AMJ7 was the most effective strain (10.13±0.2mm), and AMJ1 and AHA5 showed more inhibitory activity against Escherichia coli (8.04±0.02 mm and 8.2±0.03 ). The AMJ6 strain showed best antibacterial activity against Klebsiella sp. (8.03±0.02mm). Antifungal activity of AMJ2 showed that it was most active strain against complex (16.05±0.02mm) and against Aspergillus flavus strain AMJ1 was most active strain (16.4±0.2mm) and highest antifungal activity against Trichophyton mentagrophytes, Microsporum gyp serum and Candida albicans, were shown by AHA1 (21.03±0.02mm), AHA3 and AHA7 (18±0.03mm), AMJ6 (21.03±0.2mm) respectively. Our results revealed that the marine actinomycetes of Persian Gulf sediments were potent source of novel antibiotics and bioactive compounds and indicated that the antimicrobial metabolites were extracellular. Most of the secondary metabolites and antibiotics are extracellular in nature and extracellular products of actinomycetes show potent antimicrobial activities.

Keywords: antibacterial activity, antifungal activity, marine actinomycetes, Persian Gulf

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Authors: Sohan Sengupta, Arnab Pramanik, Abhrajyoti Ghosh, Maitree Bhattacharyya

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Newly emerged phyto-pathogens and multi drug resistance have been threating the world for last few decades. Actinomycetes, the most endowed group of microorganisms isolated from unexplored regions of the world may be the ultimate solution to these problems. Thus the aim of this study was to isolate several bioactive actinomycetes strains capable of producing antimicrobial secondary metabolite from Sundarbans, the only mangrove tiger land of the world. Fifty four actinomycetes were isolated and analyzed for antimicrobial activity against fifteen test organisms including three phytopathogens. Nine morphologically distinct and biologically active isolates were subjected to polyphasic identification study. 16s rDNA sequencing indicated eight isolates to reveal maximum similarity to the genus streptomyces, whereas one isolate presented only 93.57% similarity with Streptomyces albogriseolus NRRL B-1305T. Seventy-one carbon sources and twenty-three chemical sources utilization assay revealed their metabolic relatedness. Among these nine isolates three specific strains were found to have notably higher degree of antimicrobial potential effective in a broader range including phyto-pathogenic fungus. PCR base whole genome screen for PKS and NRPS genes, confirmed the occurrence of bio-synthetic gene cluster in some of the isolates for novel antibiotic production. Finally the strain SMS_SU21, which showed antimicrobial activity with MIC value of 0.05 mg ml-1and antioxidant activity with IC50 value of 0.242±0.33 mg ml-1 was detected to be the most potential one. True prospective of this strain was evaluated utilizing GC-MS and the bioactive compound responsible for antimicrobial activity was purified and characterized. Rare bioactive actinomycetes were isolated from unexplored heritage site. Diversity of the biosynthetic gene cluster for antimicrobial compound production has also been evaluated. Antimicrobial compound SU21-C has been identified and purified which is active against a broad range of pathogens.

Keywords: actinomycetes, sundarbans, antimicrobial, pks nrps, phyto-pathogens, GC-MS

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Authors: Kottaridi Klimentia, Demopoulos Vasilis, Sidiropoulos Anastasios, Ihara Diego, Nikolaidis Vasileios, Antonopoulos Dimitrios

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Aflatoxins are highly poisonous and carcinogenic compounds produced by species of the genus Aspergillus spp. that can infect a variety of agricultural foods, including dried figs. Biological and environmental factors, such as population, pathogenicity, and aflatoxinogenic capacity of the strains, topography, soil, and climate parameters of the fig orchards, are believed to have a strong effect on aflatoxin levels. Existing methods for aflatoxin detection and measurement, such as high performance liquid chromatography (HPLC), and enzyme-linked immunosorbent assay (ELISA), can provide accurate results, but the procedures are usually time-consuming, sample-destructive, and expensive. Predicting aflatoxin levels prior to crop harvest is useful for minimizing the health and financial impact of a contaminated crop. Consequently, there is interest in developing a tool that predicts aflatoxin levels based on topography and soil analysis data of fig orchards. This paper describes the development of a risk assessment tool for the contamination of aflatoxin on dried figs, based on the location and altitude of the fig orchards, the population of the fungus Aspergillus spp. in the soil, and soil parameters such as pH, saturation percentage (SP), electrical conductivity (EC), organic matter, particle size analysis (sand, silt, clay), the concentration of the exchangeable cations (Ca, Mg, K, Na), extractable P, and trace of elements (B, Fe, Mn, Zn and Cu), by employing machine learning methods. In particular, our proposed method integrates three machine learning techniques, i.e., dimensionality reduction on the original dataset (principal component analysis), metric learning (Mahalanobis metric for clustering), and k-nearest neighbors learning algorithm (KNN), into an enhanced model, with mean performance equal to 85% by terms of the Pearson correlation coefficient (PCC) between observed and predicted values.

Keywords: aflatoxins, Aspergillus spp., dried figs, k-nearest neighbors, machine learning, prediction

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Authors: Arun Joshi

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The present study deals with the report of Gangamopteriscyclopteroides from the Barakar Formation of Simlong Open Cast Mine, Rajmahal Area, Rajmahal Basin, Jharkhand, India. The genus Gangamopteriscomprises leaves which are simple, entire, symmetrical or asymmetrical, linear, lanceolate, elliptical, obovate in shape, apex broadly rounded, obtuse, acute, acuminate or mucronate, base petiolate or contracted, midrib absent. Median region occupied by subparallel veins with anastomoses of elongate or hexagonal outline. Secondary veins arise from median veins by repeated dichotomy, arched, bifurcating and anasotomosing network. The present work is significant as it represents the presence of Glossopteris flora (250- 290 ma) which is mainly responsible for the formation of coal. Coal is one of the major fuels for power production through thermal power plants. The Glossopteris flora is one of the major floras that occupied the southern continent during Carboniferous- Permian time. This southern continent is also known as Gondwana comprising Australia, South Africa, Antarctica, Madagascar and India. There is a vast geological reserve of coal with favorable stripping ratio available at the Simlong Block but the area comes under the most naxalite prone area and thus the mine has been running in an unplanned manner. It has got the potential of becoming a big project with higher capacity and is well suited for enhancing production which can be helpful in the economic growth of the country. Though, the present record is scanty, it shows the presence of Glossopteris flora responsible for the formation of coal in the Coalmine. However, there are fears of fossils disappearing from this area as the state government of Jharkhand has given out a mining lease in the area to private companies. Therefore, it is very necessary to study such coal forming vegetation and their systematic study from the area to generate a new palaeobotanical database, palaeoenvironmental interpretation, basinal correlation and for the understanding of evolutionary perspectives.

Keywords: Barakar formation, coal, Glossopteris flora, Gondwana, India, Naxalite, Rajmahal Basin

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Authors: Mukolwe D. Lubembe, Odongo O. David, Githaka Naftali, Kanduma Esther, Marinda Oosthuizen, Kgomotso P. Sibeko

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Theileriosis is a tick-borne disease of cattle caused by an apicomplexan protozoan parasite of the genus Theileria. In eastern and southern Africa, Theileria infections in cattle are caused by the species Theileria parva whose natural reservoir is the African buffalo (Syncerus caffer). Currently, East Coast Fever (ECF) caused by the cattle-derived Theileria parva is still a major problem in eastern Africa and some parts of southern Africa but not in South Africa following its eradication in the 1950s. However, Corridor disease (CD) caused by the buffalo-derived Theileria parva still remains a concern in South Africa. The diversity of Theileria parva in South Africa in comparison to other affected countries is poorly defined yet its known to be the survival strategy of this parasite. We assessed the antigenic diversity of Theileria parva isolates from Buffalo and cattle at the wildlife-livestock interface comparing samples from South Africa, Zimbabwe, Kenya, Tanzania, and Uganda. Antigenic epitopes of eight schizont antigen genes (Tp1, Tp3, Tp4, Tp5, Tp6, Tp7, Tp8 and Tp10) were amplified by PCR from genomic DNA extracted from blood samples collected from cattle and buffalo at the wildlife-livestock interface. Amplicons were purified and then sequenced on NGS platform. Full length open reading frames (ORFs) of two schizont antigen genes (Tp2 and Tp9) and one sporozoite antigen gene, p67 were also amplified from genomic DNA. Amplicons were then purified and cloned for sequencing. Analysis was based on sequence differences in the genes. Preliminary results show an extensively diverse population of Theileria parva circulating in buffalo and cattle populations at the wildlife-livestock interface. Diversity of the antigen genes contributes to the evasion of the immune system of the host by Theileria parva. This possess a concern in that, some of the Theileria parva populations may re-assort and become adapted to cattle to cause a form of theileriosis that is as fatal as ECF in areas where ECF was eradicated or is absent

Keywords: Theileria parva, east coast fever, corridor diseases, antigen genes, diversity

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Authors: Subhankar Kumar Sarkar

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Beetles of the subfamily Dynastinae under the family Scarabaeidae of the insect order Coleoptera are popularly known as ‘Rhinoceros beetles’ because of the characteristic horn borne by the males on their head. These horns are dedicated in mating battle against other males and have evolved as a result of phenotypic plasticity. Scarabaeidae is the largest of all families under Coleoptera and is composed of 11 subfamilies, of which the subfamily Dynastinae is represented by approximately 300 species. Some of these beetles have been reported to cause considerable damage to agriculture and forestry both in their larval and adult stages, while many of them are beneficial as they pollinate plants and recycle plant materials. Eastern Himalayas is regarded as one of the 35 biodiversity hotspot zones of the world and one of the four of India, which is exhibited by its rich and megadiverse tropical forests. However, our knowledge on the faunal diversity of these forests is very limited, particularly for the insect fauna. One such tropical forest of Eastern Himalayas is the ‘Buxa Tiger Reserve’ located between latitudes 26°30” to 26°55” North and Longitudes 89°20” to 89˚35” East of India and occupies an area of about 759.26 square kilometers. It is with this background an attempt has been made to explore the insect fauna of the forest. Insect sampling was carried out in each beat and range of Buxa Tiger Reserve in all the three seasons viz, Premonsoon, Monsoon, and Postmonsoon. Sample collections were done by sweep nets, hand picking technique and pit fall traps. UV light trap was used to collect the nocturnal insects. Morphological examinations of the collected samples were carried out with Stereozoom Binocular Microscopes (Zeiss SV6 and SV11) and were identified up to species level with the aid of relevant literature. Survey of the insect fauna of the forest resulted in the recognition of 76 scarab species, of which 8 belong to the subfamily dealt herein. Each of the 8 species represents a separate genus. The forest is dominated by the members of Xylotrupes gideon (Linnaeus) as is represented by highest number of individuals. The recorded taxa show about 12% endemism and are of mainly oriental in distribution. Premonsoon is the most favorable season for their occurrence and activity followed by Monsoon and Postmonsoon.

Keywords: Dynastinae, Scarabaeidae, diversity, Buxa Tiger Reserve

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Authors: Cha-ah C. N., Awah N. J., Mouiche M. M. M.

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Brucellosis is a neglected zoonotic disease of animals and man caused by bacteria of genus Brucella. Though eradicated in some parts of the world, it remains endemic in sub-Saharan Africa including Cameroon. The aim of this study was to contribute to the epidemiology of brucellosis in the North-West region of Cameroon by detecting the presence of anti-Brucella antibodies in bovine bulk milk as this serves as a route of transmission from animals to man. A cross sectional study was conducted to determine the prevalence of Brucella abortus antibodies in bovine bulk milk in the peri-urban zones of Bamenda. One hundred bulk milk samples were collected from 100 herds and tested by milk I-ELISA test. The conducted study revealed the presence of anti-Brucella abortus antibodies in bovine bulk milk. The study revealed that bovine brucellosis is widespread in animal production systems in this area. The animal infection pressure in these systems has remained strong due to movement of livestock in search of pasture, co-existence of animal husbandry, communal sharing of grazing land, concentration of animals around water points, abortions in production systems, locality of production systems and failure to quarantine upon introduction of new animals. The circulation of Brucella abortus antibodies in cattle farms recorded in the study revealed potential public health implication and suggest economic importance of brucellosis to the cattle industry in the Northwest region of Cameroon. The risk for re-emergence and transmission of brucellosis is evident as a result of the co-existence of animal husbandry activities and social-cultural activities that promote brucellosis transmission. Well-designed countrywide, evidence-based studies of brucellosis are needed. These could help to generate reliable frequency and potential impact estimates, to identify Brucella reservoirs, and to propose control strategies of proven efficacy.

Keywords: brucellosis, bulk milk, northwest region Cameroon, prevalence

Procedia PDF Downloads 147

Authors: Guna Raj Dhungana, Roshan Nepal, Apshara Parajuli, , Archana Maharjan, Shyam K. Mishra, Pramod Aryal, Rajani Malla

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Introduction: Klebsiella pneumoniae is a well-known opportunistic human pathogen, primarily causing healthcare-associated infections. The global emergence of carbapenemase-producing K. pneumoniaeis a major public health burden, which is often extensively multidrug resistant.Thus, because of the difficulty to treat these ‘superbug’ and menace and some term as ‘apocalypse’ of post antibiotics era, an alternative approach to controlling this pathogen is prudent and one of the approaches is phage mediated control and/or treatment. Objective: In this study, we aimed to isolate novel bacteriophage against carbapenemase-producing K. pneumoniaeand characterize for potential use inphage therapy. Material and Methods: Twenty lytic phages were isolated from river water using double layer agar assay and purified. Biological features, physiochemical characters, burst size, host specificity and activity spectrum of phages were determined. One most potent phage: Phage TU_Kle10O was selected and characterized by electron microscopy. Whole genome sequences of the phage were analyzed for presence/absence of virulent factors, and other lysin genes. Results: Novel phage TU_Kle10O showed multiple host range within own genus and did not induce any BIM up to 5th generation of host’s life cycle. Electron microscopy confirmed that the phage was tailed and belonged to Caudovirales family. Next generation sequencing revealed its genome to be 166.2 Kb. bioinformatical analysis further confirmed that the phage genome ‘did not’ contain any ‘bacterial genes’ within phage genome, which ruled out the concern for transfer of virulent genes. Specific 'lysin’ enzyme was identified phages which could be used as 'antibiotics'. Conclusion: Extensively multidrug resistant bacteria like carbapenemase-producing K. pneumoniaecould be treated efficiently by phages.Absence of ‘virulent’ genes of bacterial origin and presence of lysin proteins within phage genome makes phages an excellent candidate for therapeutics.

Keywords: bacteriophage, Klebsiella pneumoniae, MDR, phage therapy, carbapenemase,

Procedia PDF Downloads 190

Authors: M. Gogniashvili, I. Maisaia, A. Kotorashvili, N. Kotaria, T. Beridze

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Three types of plasmon (A, B and G) is typical for genus Triticum. In polyploid species - Triticum turgidum L. and Triticum aestivum L. plasmon B is detected. In the forthcoming paper, complete nucleotide sequence of chloroplast DNA of 11 representatives of Georgian wheat polyploid species, carrying plasmon B was determined. Sequencing of chloroplast DNA was performed on an Illumina MiSeq platform. Chloroplast DNA molecules were assembled using the SOAPdenovo computer program. All contigs were aligned to the reference chloroplast genome sequence using BLASTN. For detection of SNPs and Indels and phylogeny tree construction computer programs Mafft and Blast were used. Using Triticum aestivum L. subsp. macha (Dekapr. & Menabde) Mackey var. paleocolchicum Dekapr. et Menabde as a reference, 5 SNPs can be identified in chloroplast DNA of Georgian endemic polyploid wheat. The number of noncoding substitutions is 2, coding substitutions - 3. In comparison with reference DNA two - 38 bp and 56 bp inversions were observed in paleocolchicum subspecies. There were six 1 bp indels detected in Georgian polyploid wheats, all of them at microsatellite stretches. The phylogeny tree shows that subspecies macha, carthlicum and paleocolchicum occupy different positions. According to the simplified scheme based on SNP and indel data, the ancestral, female parent of the all studied polyploid wheat is unknown X predecesor, from which four lines were formed. 1 SNP and two inversions (38 bp and 56 bp) caused the formation of subsp. paleocolchicum. Three other lines are macha, durum and carthlicum lines. Macha line is further divided into two sublines (M_1 and M_4). Carthlicum line includes subsp.carthlicum and T.aestivum - C_1 - C_2 - A_1. One of the central question of wheat domestication is which people(s) participated in wheat domestication? It is proposed that the predecessors of Georgian peoples (Proto-Kartvelians) must be placed, on the evidence of archaic lexical and toponymic data, in the mountainous regions of the western and central part of the Little Caucasus (the Transcaucasian foothills) at least 4,000 years ago. One of the possibility to explain the ‘wheat puzzle’ is that Kartvelian speakers brought domesticated wheat species and subspecis from Fertile Crescent further north to South Caucasus.

Keywords: chloroplast DNA, sequencing, SNP, triticum

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Authors: Gulseren Kirbas, Mushap Kuru, Buket Bakir, Ebru Karadag Sari

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Capra (mountain goat) is a genus comprising nine species. The domestic goat (C. aegagrus hircus) is a subspecies of the wild goat that is domesticated. This study aimed to determine the anatomical structure of the salpinx and ovary of the Anatolian wild goat (C. aegagrus aegagrus). Animals that were taken to the Kafkas University Wildlife Rescue and Rehabilitation Center, Kars, Turkey, because of various reasons, such as traffic accidents and firearm injuries, were used in this study. The salpinges and ovaries of four wild goats of similar ages, which could not be rescued by the Center despite all interventions, were dissected. Measurements were taken from the right-left salpinx and ovary using digital calipers. The weights of each ovary and salpinx were measured using a precision scale (min: 0.0001 g − max: 220 g, code: XB220A; Precisa, Swiss). The histological structure of the tissues was examined after weighing the organs. The tissue samples were fixed in 10% formaldehyde for 24 h. Then a routine procedure was applied, and the tissues were embedded in paraffin. Mallory’s modified triple staining was used to demonstrate the general structure of the salpinx. The salpinx was found to consist of three different regions (infundibulum, ampulla, and isthmus). These regions consisted of tunica mucosa, tunica muscularis, and tunica serosa. The prismatic epithelial cells were observed in the lamina epithelialis of tunica mucosa in every region, but the prismatic fimbrae cells occurred most in the infundibulum. The ampulla was distinguished by its many mucosal folds. It was the longest region of the salpinx and was joined to the isthmus via the ampullary–isthmus junction. Isthmus was the caudal end of the salpinx joined to the uterus and had the thickest tunica muscularis compared with the other regions. The mean length of the ovary was 13.22 ± 1.27 mm, width was 8.46 ± 0.88 mm, the thickness was 5.67 ± 0.79 mm, and weight was 0.59 ± 0.17 g. The average length of the salpinx was 58.11 ± 14.02 mm, width was 0.80 ± 0.22 mm, the thickness was 0.41 ± 0.01 mm, and weight was 0.30 ± 0.08 g. In conclusion, the Anatolian wild goat, which is included in wildlife diversity in Turkey, has been disappearing due to illegal and uncontrolled hunting as well as traffic accidents in recent years. These findings are believed to contribute to the literature.

Keywords: Anatolian wild goat, anatomy, ovary, salpinx

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Authors: Sabrina Sebbane, Alina Elena Parvu

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Introduction: Santolina rosmarinifolia is a plant of the Santolina genus, a family made of medicinal plants widely used. Some of the Santolina species have been proven to have potent anti-inflammatory and anti-oxidant effects. However, no in vivo study has been made to demonstrate this in Santolina rosmarinifolia. The aim of our study is to experimentally evaluate the potential anti-inflammatory and anti-oxidant effects of Santolina rosmarinifolia plant extracts on acute inflammation in rats. These effects are defined by measuring the modifications on nitric oxide, reactive oxygen species and anti-oxidant response in serum. Materials and Methods: Rats were divided into 5 groups (n=6). Three groups were given Santolina rosmarinifolia extract by gavage in different concentrations(100%, 50%, 25%) for a week. Inflammation was induced by i.m injection of turpentine oil on the 8th day. One group was only given turpentine oil and the fifth group acted as control and was given only saline solution. Blood was collected and serum separated. Global tests were used to measure the oxidative stress, total oxidative status (TOS), total antioxidant reactivity (TAR) and the modified method of Griess assay to measure NO synthesis. Malondilaldehyde (MDA) and thiols levels were also assessed. Results: Santolina rosmarinifolia did not significantly change the TOS levels (p > 0.05). Santolina rosmarinifolia 25% and 50% decreased significantly the TAR levels (p < 0.001). Santolina 100% didn't have a significant effect on TAR (p > 0.05). All concentrations of Santolina rosmarinifolia increased the oxidative stress index (OSI) significantly(p < 0.05). Santolina rosmarinifolia 100% significantly decreased NO synthesis (p value < 0.05). In the diluted Santolina groups, no significant effect on NO synthesis was observed. In the groups treated with Santolina 100% and Santolina rosmarinifolia 50%, thiols concentration were significantly higher compared to the inflammation group (p < 0.02). A higher stimulatory effect was found in the Santolina 25% group (p value < 0.05). MDA levels were not significantly modified by the administration of Santolina rosmarinifolia (p > 0.05). Conclusion: All three solutions of Santolina rosmarinifolia had no important effect on oxidant production. However, Santolina rosmarinifolia solutions had a positive effect by increasing the thiols concentration in the serum of the models. The sum of all the effects produced by the administration of Santolina did not show a significant decrease of nitro-oxidative stress. Further experiments including smaller concentrations of Santolina rosmarinifolia will be made. Santolina rosmarinifolia should also be tested as a curative treatment.

Keywords: inflammation, MDA, nitric oxide, santolina rosmarinifolia, thiols, TAR, TOS

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Authors: Sunghee Nam

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Entomopathogenic fungi is a Cordyceps species that is isolated from dead silkworm and cicada. Fungi on cicadas were described in old Chinese medicinal books and from ancient times, vegetable wasps and plant worms were widely known to have active substance and have been studied for pharmacological use. Among many fungi belonging to the genus Cordyceps, Cordyceps sinensis have been demonstrated to yield natural products possessing various biological activities and many bioactive components. Generally, It is commonly used to replenish the kidney and soothe the lung, and for the treatment of fatigue. Due to their commercial and economic importance, the demand for Cordyceps has been rapidly increased. However, a supply of Cordyceps specimen could not meet the increasing demand because of their sole dependence on field collection and habitat destruction. Because it is difficult to obtain many insect hosts in nature and the edibility of host insect needs to be verified in a pharmacological aspect. Recently, this setback was overcome that P. tenuipes was able to be cultivated in a large scale using silkworm as host. Pharmacological effects of P. tenuipes cultured on silkworm such as strengthening immune function, anti-fatigue, anti-tumor activity and controlling liver etc. have been proved. They are widely commercialized. In this study, we attempted to establish a method for stable growth inhibition of P. tenuipes on silkworm hosts and an optimal condition for synnemata formation. To determine optimum culturing conditions, temperature and light conditions were varied. The length and number of synnemata was highest at 25℃ temperature and 100~300 lux illumination. On an average, the synnemata of wild P. tenuipes measures 70 ㎜ in length and 20 in number; those of the cultured strain were relatively shorter and more in number. The number of synnemata may have increased as a result of inoculating the host with highly concentrated conidia, while the length may have decreased due to limited nutrition per individual. It is not able that changes in light illumination cause morphological variations in the synnemata. However, regulation of only light and temperature could not produce stromata like perithecia, asci, and ascospores.

Keywords: optimization of culture conditions of paecilomyces tenuipes, entomopathogenic fungi optimization of culture conditions of paecilomyces tenuipes, entomopathogenic fungi silkworm larva, bombyx mori

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Authors: Danutė Jablonskytė-Raščė, Vidas Damanauskas

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The wheat (Triticum) genus is divided into many species, of which only two are widely distributed in the world - common wheat (Triticum aestivum L.) and durum wheat (Triticum durum Desf.). Common (soft) wheat is the most common type of wheat in the world and the most suitable for the harsh climate of Lithuania, but the grains have lower protein content and poorer nutritional properties. Durum wheat is characterized by a high protein content of the grain, but it is a crop of warmer climates grown in southern countries, Italy, Spain, the United States, Egypt, etc. Today's important issue is food, its resources and quality. The research focuses on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the warming climate conditions. Climatic conditions change the distribution of fungi and their hosts. Plants that have grown in our climate for many years have adapted to the use of fungicides, so the aim is to study cereal varieties grown in warmer climates and compare them with our country's varieties, studying their nutritional value and the spread of fungal diseases. The field experiments of different varieties of wheat were conducted at Joniškėlis Experimental Station of the Lithuanian Research Centre for Agriculture and Forestry in 2023. The soil of the experimental site was Endocalcari-Endohypogleyic Cambisol (CMg-n-w-can). The research was designed to identify the resistance to leaf diseases and the nutritional value of various wheat varieties. This research aims to focus on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the conditions of the warming climate. The study found that hot and humid summer weather led to the spread of foliar diseases in wheat. Tan spot (Pyrenophora tritici-repentis) is mostly spread in wheat crops. This disease had an average prevalence of 86.90%. The wheat crop was sparse, so this year was unfavorable for the spread of powdery mildew (Blumeria graminis). Dry weather prevailed during the period of flowering of cereals, which prevented the spread of ear diseases. Examining the qualitative indicators of grain, it was found that durum wheat had the best parameters.

Keywords: varieties, wheat, leaf disease, grain quality

Procedia PDF Downloads 46

Authors: Engin Berber, Nurettin Canakoglu, Ibrahim Sozdutmaz, Merve Caliskan, Shaikh Terkis Islam Pavel, Hazel Yetiskin, Aykut Ozdarendeli

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Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus in the family Bunyaviridae, genus Nairovirus. The CCHFV genome consists of three molecules of negative-sense single-stranded RNA, each encapsulated separately. The virion particle contains viral RNA polymerase (L segment), surface glycoproteins Gn and Gc (Msegment), and a nucleocapsid protein NP (S segment). CCHF is characterized by high case mortality, occurring in Asia, Africa, the Middle East and Eastern Europe. Clinical CCHF was first recognized in Turkey in 2002. The numbers of CCHF cases have gradually increased in Turkey making the virus a public health concern. Between 2002 and 2014, more than 8000 the CCHF cases have been reported in Turkey and mortality rate is around 5%. So, Turkey is one of the countries where the epidemy has become spread to the wider geography and the biggest outbreaks of CCHF have occurred in the world. We have recently developed an inactivated cell-culture based vaccine against CCHF. We have showed that the Balb/c mice immunized with the CCHF vaccine induced the high level of neutralizing antibodies. In this study, we aimed to determine the immunodominant regions of nucleoprotein (NP) CCHFV Kelkit06 strain which stimulate T cells. For this purpose, pools of overlapping NP were used for an IFN- γ ELISPOT assay. Balb/c mice were divided into two groups for the experiment. Two groups (n = 10 each) were immunized via the intraperitoneal route with 5, or 10μg of the cell culture-based vaccine. The control group (n = 6) was mock immunized with PBS. Booster injections with the same formulation were given on days 21 and 42 after the first immunization. The higher reactivity against the CCHFV NP pools 31-40 and 80-90 was determined in the two dose groups. In order to analyze the vaccine-induced T cell responses in Balb/c mice immunized with varying doses of the vaccine, we have been also currently working on CD4+, CD8+ and CD3 + T cells by flow cytometry.

Keywords: Crimean-Congo hemorrhagic fever virus, immunodominant regions of NP, T cell response, vaccine

Procedia PDF Downloads 346

Authors: Amanda J. Burridge, Keith J. Edwards, Paul A. Wilkinson, Tom Batstone, Erik H. Murchie, Lorna McAusland, Ana Elizabete Carmo-Silva, Ivan Jauregui, Tracy Lawson, Silvere R. M. Vialet-Chabrand

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The rate of genetic progress in wheat production must be improved to meet global food security targets. However, past selection for domestication traits has reduced the genetic variation in modern wheat cultivars, a fact that could severely limit the future rate of genetic gain. The genetic variation in agronomically important traits for the wild relatives and progenitors of wheat is far greater than that of the current domesticated cultivars, but transferring these traits into modern cultivars is not straightforward. Between the elite cultivars of wheat, photosynthetic capacity is a key trait for which there is limited variation. Early screening of wheat wild relative and progenitors has shown differences in photosynthetic capacity and efficiency not only between wild relative species but marked differences between the accessions of each species. By identifying wild relative accessions with improved photosynthetic traits and characterising the genetic variation responsible, it is possible to incorporate these traits into advanced breeding programmes by wide crossing and introgression programmes. To identify the potential variety of photosynthetic capacity and efficiency available in the secondary and tertiary genepool, a wide scale survey was carried out for over 600 accessions from 80 species including those from the genus Aegilops, Triticum, Thinopyrum, Elymus, and Secale. Genotype data were generated for each accession using a ‘Wheat Wild Relative’ Single Nucleotide Polymorphism (SNP) genotyping array composed of 35,000 SNP markers polymorphic between wild relatives and elite hexaploid wheat. This genotype data was combined with phenotypic measurements such as gas exchange (CO₂, H₂O), chlorophyll fluorescence, growth, morphology, and RuBisCO activity to identify potential breeding material with enhanced photosynthetic capacity and efficiency. The data and associated analysis tools presented here will prove useful to anyone interested in increasing the genetic diversity in hexaploid wheat or the application of complex genotyping data to plant breeding.

Keywords: wheat, wild relatives, pre-breeding, genomics, photosynthesis

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Authors: Aleya Ferdausi, Meriel Jones, Anthony Halls

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The Amaryllidaceae genus Narcissus contains secondary metabolites, which are important sources of bioactive compounds such as pharmaceuticals indicating that their biological activity extends from the native plant to humans. Transcriptome analysis (RNA-seq) is an effective platform for the identification and functional characterization of candidate genes as well as to identify genes encoding uncharacterized enzymes. The biotechnological production of secondary metabolites in plant cell or organ cultures has become a tempting alternative to the extraction of whole plant material. The biochemical pathways for the production of secondary metabolites require primary metabolites to undergo a series of modifications catalyzed by enzymes such as cytochrome P450s, methyltransferases, glycosyltransferases, and acyltransferases. Differential gene expression analysis of Narcissus was obtained from two conditions, i.e. field and in vitro callus. Callus was obtained from modified MS (Murashige and Skoog) media supplemented with growth regulators and twin-scale explants from Narcissus cv. Carlton bulb. A total of 2153 differentially expressed transcripts were detected in Narcissus bulb and in vitro callus, and 78.95% of those were annotated. It showed the expression of genes involved in the biosynthesis of alkaloids were present in both conditions i.e. cytochrome P450s, O-methyltransferase (OMTs), NADP/NADPH dehydrogenases or reductases, SAM-synthetases or decarboxylases, 3-ketoacyl-CoA, acyl-CoA, cinnamoyl-CoA, cinnamate 4-hydroxylase, alcohol dehydrogenase, caffeic acid, N-methyltransferase, and NADPH-cytochrome P450s. However, cytochrome P450s and OMTs involved in the later stage of Amaryllidaceae alkaloids biosynthesis were mainly up-regulated in field samples. Whereas, the enzymes involved in initial biosynthetic pathways i.e. fructose biphosphate adolase, aminotransferases, dehydrogenases, hydroxyl methyl glutarate and glutamate synthase leading to the biosynthesis of precursors; tyrosine, phenylalanine and tryptophan for secondary metabolites were up-regulated in callus. The knowledge of probable genes involved in secondary metabolism and their regulation in different tissues will provide insight into the Narcissus plant biology related to alkaloid production.

Keywords: narcissus, callus, transcriptomics, secondary metabolites

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Authors: Nilgün Öztürk, Hakan Sabahtin Ali, Hülya Tuba Kıyan

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The genus Chenopodium belongs to Amaranthaceae, is represented by approximately 250 species in the world and 15 species and three subspecies in Turkey. Chenopodium species are traditionally used to treat chest and abdominal pain, shortness of breath, cough and neurological disorders. Chenopodium quinoa Willd. (Quinoa) is native to Andes region of South America (especially Peru and Bolivia) and cultivated in many countries include also Turkey in the world nowadays. The seeds of quinoa are rich in protein, and the phytochemical composition consists of antioxidant substances such as polyphenolic compounds, flavonoids, vitamins, and minerals; anticancer and neuroprotective compounds such as tocotrienols; anti-inflammatory compounds such as carotenoids and anthocyanins and also saponins and starch. Food products of quinoa such as quinoa cereal bar, pasta and cornflakes are used in the diet made during many disorders like obesity, cardiovascular disorder, hypertension and Celiac disease. Also quinoa seems to have antimicrobial, anti-inflammatory and cholesterol-lowering properties because of its bioactive compounds. In this present study, the aqueous ethanolic extracts of the seeds of three different coloured genotypes of quinoa were investigated for their antioxidant activities using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, ferrous ion-chelating effect, ferric-reducing antioxidant power, ABTS radical cation decolorization assays and total phenolic contents using Folin-Ciocalteu assay. Among the three genotypes of quinoa; the aqueous ethanolic extract of the red genotype had the highest total phenolic content (83.54 ± 2.12 mg gallic acid/100 g extract) whereas the extract of the white genotype had the lowest total phenolic content (70.66 ± 0.25 mg gallic acid/100 g). According to the antioxidant activity results; the extracts showed moderate reducing power effect whereas weak ABTS radical cation decolorization and ferrous ion-chelating effect and also too weak DPPH radical scavenging activity when compared to the positive standards.

Keywords: amaranthaceae, antioxidant activity, Chenopodium quinoa willd., total phenolic content

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Authors: T. Urushadze, R. Khvedelidze, L. Kutateladze, M. Jobava, T. Burduli, T. Alexidze

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There is an increasing interest in reliable, wasteless, ecologically friendly technologies. About 40% of enzymes produced all over the world are used for production of syrups with high concentration of glucose-fructose. One of such technologies complies obtaining fermentable sugar glucose from raw materials containing starch by means of amylases. In modern alcohol-producing factories this process is running in two steps, involving two enzymes of different origin: bacterial α-amylase and fungal glucoamylase, as generally fungal amylases are less thermostable as compared to bacterial amylases. Selection of stable and operable at 700С and higher temperatures enzyme preparation with both α- and glucoamylase activities will allow conducting this process in one step. S. Durmishidze Institute of Biochemistry and Biotechnology owns unique collection of mycelial fungi, isolated from different ecological niches of Caucasus. As a result of screening our collection 39 strains poducing amylases were revealed. Most of them belong to the genus Aspergillus. Optimum temperatures of action of selected amylases from three producers were estableshed to be within the range 67-80°C. A. niger B-6 showed higher α-amylase activity at 67°C, and glucoamylase activity at 62°C, A. niger 6-12 showed higher α-amylase activity at 72°C, and glucoamylase activity at 65°C, Aspergillus niger p8-3 showed higher activities at 82°C and 70°C, for α-amylase and glucoamylase activities, respectively. Exhaustive hydrolysis process of starch solutions of different concentrations (3, 5, 15, and 30 %) with cultural liquid and technical preparation of Aspergillus niger p8-3 enzyme was studied. In case of low concentrations exhaustive hydrolysis of starch lasts 40–60 minutes, in case of high concentrations hydrolysis takes longer time. 98, 6% yield of glucose can be reached at incubation during 12 hours with enzyme cultural liquid and 8 hours incubation with technical preparation of the enzyme at gradual increase of temperature from 50°C to 82°C during the first 20 minutes and further decrease of temperature to 70°C. Temperature setting for high yield of glucose and high hydrolysis (pasteurizing), optimal for activity of these strains is the prerequisite to be able to carry out hydrolysis of starch to glucose in one step, and consequently, using one strain, what will be economically justified.

Keywords: amylase, glucose hydrolisis, stability, starch

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Authors: Nisha Singh, Munish Puri, Collin Barrow, Deepak Tuli, Anshu S. Mathur

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The biological conversion of lignocellulosic biomass to ethanol is a promising strategy to solve the present global crisis of exhausting fossil fuels. The existing bioethanol production technologies have cost constraints due to the involvement of mandate pretreatment and extensive enzyme production steps. A unique process configuration known as consolidated bioprocessing (CBP) is believed to be a potential cost-effective process due to its efficient integration of enzyme production, saccharification, and fermentation into one step. Due to several favorable reasons like single step conversion, no need of adding exogenous enzymes and facilitated product recovery, CBP has gained the attention of researchers worldwide. However, there are several technical and economic barriers which need to be overcome for making consolidated bioprocessing a commercially viable process. Finding a natural candidate CBP organism is critically important and thermophilic anaerobes are preferred microorganisms. The thermophilic anaerobes that can represent CBP mainly belong to genus Clostridium, Caldicellulosiruptor, Thermoanaerobacter, Thermoanaero bacterium, and Geobacillus etc. Amongst them, Clostridium thermocellum has received increased attention as a high utility CBP candidate due to its highest growth rate on crystalline cellulose, the presence of highly efficient cellulosome system and ability to produce ethanol directly from cellulose. Recently with the availability of genetic and molecular tools aiding the metabolic engineering of Clostridium thermocellum have further facilitated the viability of commercial CBP process. With this view, we have specifically screened cellulolytic and xylanolytic thermophilic anaerobic ethanol producing bacteria, from unexplored hot spring/s in India. One of the isolates is a potential CBP organism identified as a new strain of Clostridium thermocellum. This strain has shown superior avicel and xylan degradation under unoptimized conditions compared to reported wild type strains of Clostridium thermocellum and produced more than 50 mM ethanol in 72 hours from 1 % avicel at 60°C. Besides, this strain shows good ethanol tolerance and growth on both hexose and pentose sugars. Hence, with further optimization this new strain could be developed as a potential CBP microbe.

Keywords: Clostridium thermocellum, consolidated bioprocessing, ethanol, thermophilic anaerobes

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

Abstract:

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

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

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