Search results for: rapid microbial identification

Authors: Rajesh P.P., Md. Tabish Noori, Makarand M. Ghangrekar

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Methanogenic substrate loss is reported to be a major bottleneck in microbial fuel cell which significantly reduces the power production capacity and coulombic efficiency (CE) of microbial fuel cell (MFC). Nitroethane is found to be a potent inhibitor of hydrogenotrophic methanogens in rumen fermentation process. Influence of nitroethane pre-treated sewage sludge inoculum on suppressing the methanogenic activity and enhancing the electrogenesis in MFC was evaluated. MFC inoculated with nitroethane pre-treated anodic inoculum demonstrated a maximum operating voltage of 541 mV, with coulombic efficiency and sustainable volumetric power density of 39.85 % and 14.63 W/m3 respectively. Linear sweep voltammetry indicated a higher electron discharge on the anode surface due to enhancement of electrogenic activity while suppressing methanogenic activity. A 63 % reduction in specific methanogenic activity was observed in anaerobic sludge pre-treated with nitroethane; emphasizing significance of this pretreatment for suppressing methanogenesis and its utility for enhancing electricity generation in MFC.

Keywords: coulombic efficiency, methanogenesis inhibition, microbial fuel cell, nitroethane

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Authors: Shreya Borthakur, Adhar Sharma

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The North-east states of India are well known for their age-old practice of preparing alcoholic beer from rice and millet. They do so in a traditional way by sprinkling starter cake (inoculum) on cooked rice or millet after which the fermentation starts and eventually, forms the beer. This starter cake has a rich composition of different microbes and medicinal herbs along with the powdered rice dough or maize dough with rice bran. The starter cake microbial composition has an important role in determining the microbial succession and metabolic secretions as the fermentation proceeds from the early to its late stage, thus, giving the beer a unique aroma, taste, and other sensory properties of traditionally prepared beer. Here, We have worked on identifying and characterizing the microbial community in the starter cakes prepared by the Monpa and Galo tribes of Arunachal Pradesh. A total of 18 microbial strains have been isolated from the starter cake of Monpa tribe, while 10 microbial isolates in that of Galo tribe. A metagenomic approach was applied to enumerate the cultural and non-cultural microbes present in the starter cakes prepared by the Monpa and Galo tribes of Arunachal Pradesh. The findings of the mini-project lays foundation to understand the role of microbes present in the starter cake in the beer’s fermentation process and will aide in future research on re-formulating the starter cakes to prevent the early spoilage of the ready to consume beer as the traditional rice beer has a short shelf-life. The paper concludes with the way forward being controlled CRISPR-Cas9.

Keywords: fermentation, traditional beer, microbial succession, preservation, CRISPR-Cas, food microbiology

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Authors: Jayesh M. Sonawane, Prakash C. Ghosh

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Landfill leachate emerges as a promising feedstock for microbial fuel cells (MFCs). In the present investigation, direct air-breathing cathode-based MFCs are fabricated to investigate the potential of landfill leachate. Three MFCs that have different cathode areas are fabricated and investigated for 17 days under open circuit conditions. The maximum open circuit voltage (OCV) is observed to be as high as 1.29 V. The maximum cathode area specific power density achieved in the reactor is 1513 mW m^-2. Further studies are under progress to understand the origin of high OCV obtained from landfill leachate-based MFCs.

Keywords: microbial fuel cells, landfill leachate, air-breathing cathode, performance study

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Authors: Zeina Bourhane, Anders Lanzen, Christine Cagnon, Olfa Ben Said, Cristiana Cravo-Laureau, Robert Duran

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The anthropogenic pressure in coastal areas increases dramatically with the exploitation of environmental resources. Biomonitoring coastal areas are crucial to determine the impact of pollutants on bacterial communities in soils and sediments since they provide important ecosystem services. However, relevant biomonitoring tools allowing fast determination of the ecological status are yet to be defined. Microbial ecology approaches provide useful information for developing such microbial monitoring tools reporting on the effect of environmental stressors. Chemical and microbial molecular approaches were combined in order to determine microbial bioindicators for assessing the ecological status of soil and river ecosystems around the Ichkeul Lake (Tunisia), an area highly impacted by human activities. Samples were collected along soil/river/lake continuums in three stations around the Ichkeul Lake influenced by different human activities at two seasons (summer and winter). Contaminant pressure indexes (PI), including PAHs (Polycyclic aromatic hydrocarbons), alkanes, and OCPs (Organochlorine pesticides) contents, showed significant differences in the contamination level between the stations with seasonal variation. Bacterial communities were characterized by 16S ribosomal RNAs (rRNA) gene metabarcoding. Although microgAMBI indexes, determined from the sequencing data, were in accordance with contaminant contents, they were not sufficient to fully explain the PI. Therefore, further microbial indicators are still to be defined. The comparison of bacterial communities revealed the specific microbial assemblage for soil, river, and lake sediments, which were significantly correlated with contaminant contents and PI. Such observation offers the possibility to define a relevant set of bioindicators for reporting the effects of human activities on the microbial community structure. Such bioindicators might constitute useful monitoring tools for the management of microbial communities in coastal areas.

Keywords: bacterial communities, biomonitoring, contamination, human impacts, microbial bioindicators

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Authors: Flackson Tshuma, James Bennett, Pieter Andreas Swanepoel, Johan Labuschagne, Stephan van der Westhuizen, Francis Rayns

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Amongst other things, tillage and synthetic agrochemicals can be effective methods of seedbed preparation and pest control. Nonetheless, frequent and intensive tillage and excessive application of synthetic agrochemicals, such as herbicides and insecticides, can reduce soil microbial enzyme activity. A decline in soil microbial enzyme activity can negatively affect nutrient cycling and crop productivity. In this study, the effects of four tillage treatments; continuous mouldboard plough; shallow tine-tillage to a depth of about 75 mm; no-tillage; and tillage rotation (involving shallow tine-tillage once every four years in rotation with three years of no-tillage), and two rates of synthetic agrochemicals (standard: with regular application of synthetic agrochemicals; and reduced: fewer synthetic agrochemicals in combination with bio-chemicals/ or bio-stimulants) on soil microbial enzyme activity were investigated between 2018 and 2020 in a typical Mediterranean climate zone in South Africa. Four different bio-stimulants applied contained: Trichoderma asperellum, fulvic acid, silicic acid, and Nereocystis luetkeana extracts, respectively. The study was laid out as a complete randomised block design with four replicated blocks. Each block had 14 plots, and each plot measured 50 m x 6 m. The study aimed to assess the combined impact of tillage practices and reduced rates of synthetic agrochemical application on soil microbial enzyme activity in a dryland cropping system. It was hypothesised that the application of bio-stimulants in combination with minimum soil disturbance will lead to a greater increase in microbial enzyme activity than the effect of applying either in isolation. Six soil cores were randomly and aseptically collected from each plot for microbial enzyme activity analysis from the 0-150 mm layer of a field trial under a dryland crop rotation system in the Swartland region. The activities of four microbial enzymes, β-glucosidase, acid phosphatase, alkaline phosphatase and urease, were assessed. The enzymes are essential for the cycling of glucose, phosphorus, and nitrogen, respectively. Microbial enzyme activity generally increased with a reduction of both tillage intensity and synthetic agrochemical application. The use of the mouldboard plough led to the least (P<0.05) microbial enzyme activity relative to the reduced tillage treatments, whereas the system with bio-stimulants (reduced synthetic agrochemicals) led to the highest (P<0.05) microbial enzyme activity relative to the standard systems. The application of bio-stimulants in combination with reduced tillage, particularly no-tillage, could be beneficial for enzyme activity in a dryland farming system.

Keywords: bio-stimulants, soil microbial enzymes, synthetic agrochemicals, tillage

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Authors: Hana Barak, Alex Sivan, Ariel Kushmaro

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Microorganisms are the most diverse and abundant life forms on Earth and account for a large portion of the Earth’s biomass and biodiversity. To date though, our knowledge regarding microbial life is lacking, as it is based mainly on information from cultivated organisms. Indeed, microbiologists have borrowed from astrophysics and termed the ‘uncultured microbial majority’ as ‘microbial dark matter’. The realization of how diverse and unexplored microorganisms are, actually stems from recent advances in molecular biology, and in particular from novel methods for sequencing microbial small subunit ribosomal RNA genes directly from environmental samples termed next-generation sequencing (NGS). This has led us to use NGS that generates several gigabases of sequencing data in a single experimental run, to identify and classify environmental samples of microorganisms. In metagenomics sequencing analysis (both 16S and shotgun), sequences are compared to reference databases that contain only small part of the existing microorganisms and therefore their taxonomy assignment may reveal groups of unknown microorganisms or origins. These unknowns, or the ‘microbial sequences dark matter’, are usually ignored in spite of their great importance. The goal of this work was to develop an improved bioinformatics method that enables more complete analyses of the microbial communities in numerous environments. Therefore, NGS was used to identify previously unknown microorganisms from three different environments (industrials wastewater, Negev Desert’s rocks and water wells at the Arava valley). 16S rRNA gene metagenome analysis of the microorganisms from those three environments produce about ~4 million reads for 75 samples. Between 0.1-12% of the sequences in each sample were tagged as ‘Unassigned’. Employing relatively simple methodology for resequencing of original gDNA samples through Sanger or MiSeq Illumina with specific primers, this study demonstrates that the mysterious ‘Unassigned’ group apparently contains sequences of candidate phyla. Those unknown sequences can be located on a phylogenetic tree and thus provide a better understanding of the ‘sequences dark matter’ and its role in the research of microbial communities and diversity. Studying this ‘dark matter’ will extend the existing databases and could reveal the hidden potential of the ‘microbial dark matter’.

Keywords: bacteria, bioinformatics, dark matter, Next Generation Sequencing, unknown

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Authors: Anza-Vhudziki Mboyi, Ilunga Kamika, Maggy Momba

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The aim of this study was to ascertain the survival limit and capability of commonly found wastewater protozoan (Aspidisca sp, Trachelophyllum sp, and Peranema sp) and bacterial (Bacillus licheniformis, Brevibacillus laterosporus, and Pseudomonas putida) species to remove COD while exposed to commercial nanomaterials under varying pH conditions. The experimental study was carried out in modified mixed liquor media adjusted to various pH levels (pH 2, 7 and 10), and a comparative study was performed to determine the difference between the cytotoxicity effects of commercial zinc oxide (nZnO) and silver (nAg) nanomaterials (NMs) on the target wastewater microbial communities using standard methods. The selected microbial communities were exposed to lethal concentrations ranging from 0.015 g/L to 40 g/L for nZnO and from 0.015 g/L to 2 g/L for nAg for a period of 5 days of incubation at 30°C (100 r/min). Compared with the absence of NMs in wastewater mixed liquor, the relevant environmental concentration ranging between 10 µg/L and 100 µg/L, for both nZnO and nAg caused no adverse effects, but the presence of 20 g of nZnO/L and 0.65 g of nAg/L significantly inhibited microbial growth. Statistical evidence showed that nAg was significantly more toxic compared to nZnO, but there was an insignificant difference in toxicity between microbial communities and pH variations. A significant decrease in the removal of COD by microbial populations was observed in the presence of NMs with a moderate correlation of r = 0.3 to r = 0.7 at all pH levels. It was evident that there was a physical interaction between commercial NMs and target wastewater microbial communities; although not quantitatively assessed, cell morphology and cell death were observed. Such phenomena suggest the high resilience of the microbial community, but it is the accumulation of NMs that will have adverse effects on the performance in terms of COD removal.

Keywords: bacteria, biological treatment, chemical oxygen demand (COD) and nanomaterials, consortium, pH, protozoan

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Authors: A. Ribeiro, N. Valério, C. Vilarinho, J. Araujo, J. Carvalho

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Res2ValHUM project involves institutions from the Spanish Autonomous Region of Galicia and the north of Portugal (districts of Porto and Braga) and has as overall objectives of promotion of composting as an process for the correct managing of organic waste, valorization of compost in different fields or applications for the constitution of products with high added value, reducing of raw materials losses, and reduction of the amount of waste throw in landfills. Three main actions were designed to achieve the objectives: development of a management tool to improve collection and residue channeling for composting, sensibilization of the population for composting and characterization of the chemical and biological properties of compost and humic and fulvic substances to envisage high-value applications of compost. Here we present the cooperative activity of Galician and northern Portuguese institutions to valorize organic waste in both regions with common socio-economic characteristics and residue management problems. Results from the creation of the resource manage tool proved the existence of a large number of agricultural wastes that could be valorized. In the North of Portugal, the wastes from maize, oats, potato, apple, grape pomace, rye, and olive pomace can be highlighted. In the Autonomous Region of Galicia the wastes from maize, wheat, potato, apple, and chestnuts can be emphasized. Regarding the isolation and identification of microbial consortia from compost samples, results proved microorganisms belong mainly to the genus Bacillus spp. Among all the species identified in compost samples, Bacillus licheniformis can be highlighted in the production of humic and fulvic acids.

Keywords: agricultural wastes, Bacillus licheniformis, Bacillus spp., humic-acids, fulvic-acids

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Authors: Nurcan Cetinkaya, Nadide Hulya Ozdemir

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The aim of the study was to investigate the effect of Saccharomyces cerevisiae (SC) live yeast culture on microbial protein supply to the small intestine in Kivircik male yearlings when fed with different ratio of forage and concentrate diets. Four Kivircik male yearlings with permanent rumen canula were used in the experiment. . The treatments were allocated to a 4x4 Latin square design. Diet I consisted of 70% alfalfa hay and 30% concentrate, Diet II consisted of 30% alfalfa hay and 70% concentrate, Diet I and II were supplemented with a SC. Daily urine was collected and stored at -20°C until analysis. Calorimetric methods were used for the determination of urinary allantoin and creatinin levels. The estimated microbial N supply to small intestine for Diets I, I+SC, II and II+SC were 2.51, 2.64, 2.95 and 3.43 g N/d respectively. Supplementation of Diets I and II with SC significantly affected the allantoin levels in µmol/W0. 75 (p<0.05). Mean creatinine values in µmol/W0. 75 and allantoin:creatinin ratios were not significantly different among diets. In conclusion, supplementation with SC live yeast culture had a significant effect on urinary allantoin excretion and microbial protein supply to small intestine in Kivircik yearlings fed with high concentrate Diet II (P<0.05). Hence urinary allantoin excretion may be used as a tool for estimating microbial protein supply in Kivircık yearlings. However, further studies are necessary to understand the metabolism of Saccharomyces cerevisiae live yeast culture with different forage: concentrate ratio in Kıvırcık Yearlings.

Keywords: allantoin, creatinin, Kivircik yearling, microbial nitrogen, Saccharomyces cerevisia

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Authors: Khaoula Bensaida, Osama Eljamal

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The electrical energy generation through Microbial Fuel Cells (MFCs) using microorganisms is a renewable and sustainable approach. It creates truly an efficient technology for power production and wastewater treatment. MFC is an electrochemical device which turns wastewater into electricity. The most important part of MFC is microbes. Nano zero-valent Iron NZVI technique was successfully applied in degrading the chemical pollutants and cleaning wastewater. However, the use of NZVI for enhancing the current production is still not confirmed yet. This study aims to confirm the effect of these particles on the current generation by using MFC. A constructed microbial fuel cell, which utilizes domestic wastewater, has been considered for wastewater treatment and bio-electricity generation. The two electrodes were connected to an external resistor (200 ohms). Experiments were conducted in two steps. First, the MFC was constructed without adding NZVI particles (Control) while at a second step, nanoparticles were added with a concentration of 50mg/L. After 20 hours, the measured voltage increased to 5 and 8mV, respectively. To conclude, the use of zero-valent iron in an MFC system can increase electricity generation.

Keywords: bacterial growth, electricity generation, microbial fuel cell MFC, nano zero-valent iron NZVI.

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Authors: Ahmed A.D. Sarhan, Chong Feng Duan, Mum Wai Yip, M. Sayuti

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Rapid Prototyping (RP) technologies enable physical parts to be produced from various materials without depending on the conventional tooling. Fused Deposition Modeling (FDM) is one of the famous RP processes used at present. Tensile strength and compressive strength resistance will be identified for different sample structures and different layer orientations of ABS rapid prototype solid models. The samples will be fabricated by a FDM rapid prototyping machine in different layer orientations with variations in internal geometrical structure. The 0° orientation where layers were deposited along the length of the samples displayed superior strength and impact resistance over all the other orientations. The anisotropic properties were probably caused by weak interlayer bonding and interlayer porosity.

Keywords: building orientation, compression strength, rapid prototyping, tensile strength

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Authors: Varsha Shinde, Belle Damodara Shenoy

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Tarballs are crude oil remnants found in oceans after long term weathering process and are a global concern since several decades as potential marine pollutant. Being complicated in structure microbial remediation of tarballs in natural environment is a slow process. They are rich in high molecular weight alkanes and poly aromatic hydrocarbons which are resistant to microbial attack and other environmental factors, therefore remain in environment for long time. However, it has been found that many bacteria and fungi inhabit on tarballs for nutrients and shelter. Many of them are supposed to be oil degraders, while others are supposed to be getting benefited by byproducts formed during hydrocarbon metabolism. Thus tarballs are forming special interesting ecological niche of microbes. This work aimed to study diversity of bacteria and fungi from tarballs and to see their potential application in crude oil degradation. The samples of tarballs were collected from Betul beach of south Goa (India). Different methods were used to isolate culturable fraction of bacteria and fungi from it. Those were sequenced for 16S rRNA gene and ITS for molecular level identification. The 16S rRNA gene sequence analysis revealed the presence of 13 bacterial genera/clades (Alcanivorax, Brevibacterium, Bacillus, Cellulomonas, Enterobacter, Klebsiella, Marinobacter, Nitratireductor, Pantoea, Pseudomonas, Pseudoxanthomonas, Tistrella and Vibrio), while the ITS sequence analysis placed the fungi in 8 diverse genera/ clades (Aspergillus, Byssochlamys, Monascus, Paecilomyces, Penicillium, Scytalidium/ Xylogone, Talaromyces and Trichoderma). All bacterial isolates were screened for oil degradation capacity. Potential strains were subjected to crude oil degradation experiment for quantification. Results were analyzed by GC-MS-MS.

Keywords: bacteria, biodegradation, crude oil, diversity, fungi, tarballs

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Authors: Isaac Quiros-Fernandez, Angel Cid-Arregui

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Tumor-reactive T cell receptors (TCRs) are being subject of intense investigation since they offer great potential in adoptive cell therapies against cancer. However, the identification of tumor-specific TCRs has proven challenging, for instance, due to the limited expansion capacity of tumor-infiltrating T cells (TILs) and the extremely low frequencies of tumor-reactive T cells in the repertoire of patients and healthy donors. We have developed an approach for rapid identification and characterization of neoepitope-reactive TCRs from the T cell repertoire of healthy donors. CD8 T cells isolated from multiple donors are subjected to a first sorting step after staining with HLA multimers carrying the peptide of interest. The isolated cells are expanded for two weeks, after which a second sorting is performed using the same peptide-HLA multimers. The cells isolated in this way are then processed for single-cell sequencing of their TCR alpha and beta chains. Newly identified TCRs are cloned in appropriate expression vectors for functional analysis on Jurkat, NK92, and primary CD8 T cells and tumor cells expressing the appropriate antigen. We have identified TCRs specifically binding HLA-A2 presenting epitopes of tumor antigens, which are capable of inducing TCR-mediated cell activation and cytotoxicity in target cancer cell lines. This method allows the identification of tumor-reactive TCRs in about two to three weeks, starting from peripheral blood samples of readily available healthy donors.

Keywords: cancer, TCR, tumor antigens, immunotherapy

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Authors: Shubhangi R. Deshmukh, Anupam B. Soni

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Clean water and electricity are vital services needed in all communities. Bio-degradation of wastewater contaminants and desalination technologies are the best possible alternatives for the global shortage of fresh water supply. Osmotic microbial fuel cell (OMFC) is a versatile technology that uses microorganism (used for biodegradation of organic waste) and membrane technology (used for water purification) for wastewater treatment and energy generation simultaneously. This technology is the combination of microbial fuel cell (MFC) and forward osmosis (FO) processes. OMFC can give more electricity and clean water than the MFC which has a regular proton exchange membrane. FO gives many improvements such as high contamination removal, lower operating energy, raising high proton flux than other pressure-driven membrane technology. Lower concentration polarization lowers the membrane fouling by giving osmotic water recovery without extra cost. In this review paper, we have discussed the principle, mechanism, limitation, and application of OMFC technology reported to date. Also, we have interpreted the experimental data from various literature on the water recovery and electricity generation assessed by a different component of OMFC. The area of producing electricity using OMFC has further scope for research and seems like a promising route to wastewater treatment.

Keywords: forward osmosis, microbial fuel cell, osmotic microbial fuel cell, wastewater treatment

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Authors: Samira Berenji Ardestani, Hamid Reza Akhavan

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Gamma irradiation greatly reduces the potential microbiological risk of fresh fruits, resulting in improved microbial safety as well as extending their shelf life. The effects of 0.5-2 kGy gamma doses on some physicochemical, microbial and sensory properties of fresh barberry fruits (Berberis vulgaris) during refrigerated storage for 40 days were evaluated. The total anthocyanin and total phenolic contents of barberry fruits decreased in a dose-dependent manner immediately after irradiation and after subsequent storage. In general, it is recommended that, according to the effect of gamma radiation on physicochemical, microbial and sensorial characteristics, doses of 1.25-2 kGy could be used.

Keywords: antioxidant property, barberry fruit, chemical properties, gamma irradiation

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Authors: Karan R. Chavan, Srivats Gopalan, Kumudini V. Marathe

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In the current work, the comparison of degradation of malathion by single species, Pseudomonas Stutzeri, and Activated Sludge/Mixed Microbial Culture is studied in a Membrane Bioreactor. Various parameters were considered to study the effect of single species degradation compared to degradation by activated sludge. The experimental results revealed 85-90% reduction in the COD of the Malathion containing synthetic wastewater. Complete reduction of malathion was observed within 24 hours in both the cases. The critical flux was 10 LMH for both the systems. Fouling propensity, Cake and Membrane resistances were calculated thus giving an insight regarding the working of Membrane Bioreactor-based on single species and activated sludge.

Keywords: fouling, membrane bioreactor, mixed microbial culture, single species

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Authors: Rula D. Alshareef, Safa S. Alqathmi, Ghadah K. Alkhouldi, Reem O. Bagabas, Farheen B. Hasan

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Safety engineering is among the most crucial considerations in any work environment. Providing mentally, physically, and environmentally safe work conditions must be the top priority of any successful organization. Company X is a local paint production company in Saudi Arabia; in a month, the factory experienced two significant accidents, which indicates that workers’ safety is overlooked. The aim of the research is to examine the risks, assess the root causes and recommend control measures that will eventually contribute to providing a safe workplace. The methodology used is sectioned into three phases, risk identification, assessment, and finally, mitigation. In the identification phase, the team used Rapid Entire Body Assessment (REBA) and National Institute for Occupational Safety and Health Lifting Index (NIOSH LI) tools to holistically establish knowledge about the current risk posed to the factory. The physical hazards in the factory were assessed in two different operations, which are mixing and filling/packaging. For the risk assessment phase, the hazards were deeply analyzed through their severity and impact. Additionally, through risk mitigation, the Rapid Entire Body Assessment (REBA) score decreased from 11 to 7, and the National Institute for Occupational Safety and Health Lifting Index (NIOSH LI) has been reduced from 5.27 to 1.85.

Keywords: ergonomics, safety, workplace risks, hazards, awkward posture, fatigue, work environment

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Authors: Ansong Richard Omari, Yosei Oikawa, Yoshiharu Fujii, Dorothea Sonoko Bellingrath-Kimura

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Renewed interest in soil management aimed at improving the productive capacity of Sub Saharan Africa (SSA) soils has called for the need to analyse the long term effect of different fertilization systems on soil. This study was conducted in two agroecological zones (i.e., Guinea Savannah (GS) and Deciduous forest (DF)) of Ghana to evaluate the impacts of long term (> 5 years) fertilization schemes on soil chemical and microbial properties. Soil samples under four different fertilization schemes (inorganic, inorganic and organic, organic, and no fertilization) were collected from 20 farmers` field in both agroecological zones. Soil analyses were conducted using standard procedures. All average soil quality parameters except extractable C, potential mineralizable nitrogen and CEC were significantly higher in DF sites compared to GS. Inorganic fertilization proved superior in soil chemical and microbial biomass especially in GS zone. In GS, soil deterioration index (DI) revealed that soil quality deteriorated significantly (−26%) under only organic fertilization system whereas soil improvement was observed under inorganic and no fertilization sites. In DF, either inorganic or organic and inorganic fertilization showed significant positive effects on soil quality. The high soil chemical composition and enhanced microbial biomass in DF were associated with the high rate of inorganic fertilization.

Keywords: deterioration index, fertilization scheme, microbial biomass, tropical agroecological zone

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Authors: Uraib Sharaha, Ahmad Salman, Eladio Rodriguez-Diaz, Elad Shufan, Klaris Riesenberg, Irving J. Bigio, Mahmoud Huleihel

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Antimicrobial drugs have played an indispensable role in controlling illness and death associated with infectious diseases in animals and humans. However, the increasing resistance of bacteria to a broad spectrum of commonly used antibiotics has become a global healthcare problem. Many antibiotics had lost their effectiveness since the beginning of the antibiotic era because many bacteria have adapted defenses against these antibiotics. Rapid determination of antimicrobial susceptibility of a clinical isolate is often crucial for the optimal antimicrobial therapy of infected patients and in many cases can save lives. The conventional methods for susceptibility testing require the isolation of the pathogen from a clinical specimen by culturing on the appropriate media (this culturing stage lasts 24 h-first culturing). Then, chosen colonies are grown on media containing antibiotic(s), using micro-diffusion discs (second culturing time is also 24 h) in order to determine its bacterial susceptibility. Other methods, genotyping methods, E-test and automated methods were also developed for testing antimicrobial susceptibility. Most of these methods are expensive and time-consuming. Fourier transform infrared (FTIR) microscopy is rapid, safe, effective and low cost method that was widely and successfully used in different studies for the identification of various biological samples including bacteria; nonetheless, its true potential in routine clinical diagnosis has not yet been established. The new modern infrared (IR) spectrometers with high spectral resolution enable measuring unprecedented biochemical information from cells at the molecular level. Moreover, the development of new bioinformatics analyses combined with IR spectroscopy becomes a powerful technique, which enables the detection of structural changes associated with resistivity. The main goal of this study is to evaluate the potential of the FTIR microscopy in tandem with machine learning algorithms for rapid and reliable identification of bacterial susceptibility to antibiotics in time span of few minutes. The UTI E.coli bacterial samples, which were identified at the species level by MALDI-TOF and examined for their susceptibility by the routine assay (micro-diffusion discs), are obtained from the bacteriology laboratories in Soroka University Medical Center (SUMC). These samples were examined by FTIR microscopy and analyzed by advanced statistical methods. Our results, based on 700 E.coli samples, were promising and showed that by using infrared spectroscopic technique together with multivariate analysis, it is possible to classify the tested bacteria into sensitive and resistant with success rate higher than 90% for eight different antibiotics. Based on these preliminary results, it is worthwhile to continue developing the FTIR microscopy technique as a rapid and reliable method for identification antibiotic susceptibility.

Keywords: antibiotics, E.coli, FTIR, multivariate analysis, susceptibility, UTI

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Authors: Isabel Lopez-Oliva, Akshay Paropkari, Shweta Saraswat, Stefan Serban, Paola de Pablo, Karim Raza, Andrew Filer, Iain Chapple, Thomas Dietrich, Melissa Grant, Purnima Kumar

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Objective: We aimed to explicate the role of the subgingival microbiome in the causal link between rheumatoid arthritis (RA) and periodontitis (PD). Methods: Subjects with/without RA and with/without PD were randomized for treatment with scaling and root planing (SRP) or oral hygiene instructions. Subgingival biofilm, gingival crevicular fluid, and serum were collected at baseline and at 3- and 6-months post-operatively. Correlations were generated between 72 million 16S rDNA sequences, immuno-inflammatory mediators, circulating antibodies to oral microbial antigens, serum inflammatory molecules, and clinical metrics of RA. The dynamics of inter-microbial and host-microbial interactions were modeled using differential network analysis. Results: RA superseded periodontitis as a determinant of microbial composition, and DAS28 score superseded the severity of periodontitis as a driver of microbial assemblages (p=0.001, ANOSIM). RA subjects evidenced higher serum anti-PPAD (p=0.0013), anti-Pg-enolase (p=0.0031), anti-RPP3, anti- Pg-OMP and anti- Pi-OMP (p=0.001) antibodies than non-RA controls (with and without periodontitis). Following SRP, bacterial networks anchored by IL-1b, IL-4, IL-6, IL-10, IL-13, MIP-1b, and PDGF-b underwent ≥5-fold higher rewiring; and serum antibodies to microbial antigens decreased significantly. Conclusions: Our data suggest a circular relationship between RA and PD, beginning with an RA-influenced dysbiosis within the healthy subgingival microbiome that leads to exaggerated local inflammation in periodontitis and circulating antibodies to periodontal pathogens and positive correlation between severity of periodontitis and RA activity. Periodontal therapy restores host-microbial homeostasis, reduces local inflammation, and decreases circulating microbial antigens. Our data highlights the importance of integrating periodontal care into the management of RA patients.

Keywords: rheumatoid arthritis, periodontal, subgingival, DNA sequence analysis, oral microbiome

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Authors: Satya Eswari Jujjavarapu, Swasti Dhagat, Lata Upadhyay, Reecha Sahu

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Silver nanoparticles have a broad range of antimicrobial and antifungal properties ranging from soaps, pastes to sterilization and drug delivery systems. These can be synthesized by physical, chemical and biological methods; biological methods being the most popular owing to their non-toxic nature and reduced energy requirements. Microbial surfactants, produced on the microbial cell surface or excreted extracellularly are an alternative to synthetic surfactants for the production of silver nanoparticles. Hence, they are also called as green molecules. Microbial lipopeptide surfactants (biosurfactant) exhibit anti-tumor and anti-microbial properties and can be used as drug delivery agents. In this study, biosurfactant was synthesized by using a strain of acillus subtilis. The biosurfactant thus produced was analysed by emulsification assay, oil spilling test, and haemolytic test. Biosurfactant-mediated silver nanoparticles were synthesised by microwave irradiation of the culture supernatant and further characterized by UV–vis spectroscopy for a range of 400-600 nm. The UV–vis spectra showed a surface plasmon resonance vibration band at 410 nm corresponding to the peak of silver nanoparticles.

Keywords: biosurfactant, Bacillus subtilis, silver nano particle, lipopeptide

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Authors: Pratthana Ammaraphitak, Piyachon Ketsuwan, Rattapoom Prommana

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Electricity production from vermicompost liquid was investigated in microbial fuel cells (MFCs). The aim of this study was to determine the performance of vermicompost liquid as a biocatalyst for electricity production by MFCs. Chemical and physical parameters of vermicompost liquid as total nitrogen, ammonia-nitrogen, nitrate, nitrite, total phosphorus, potassium, organic matter, C:N ratio, pH, and electrical conductivity in MFCs were studied. The performance of MFCs was operated in open circuit mode for 7 days. The maximum open circuit voltage (OCV) was 0.45 V. The maximum power density of 5.29 ± 0.75 W/m² corresponding to a current density of 0.024 2 ± 0.0017 A/m² was achieved by the 1000 Ω on day 2. Vermicompost liquid has efficiency to generate electricity from organic waste.

Keywords: vermicompost liquid, microbial fuel cell, nutrient, electricity production

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Authors: Carlos Alberto Riveros, Edwin Fabián García, Javier Enrique Rivero

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The measured data obtained from sensors in continuous monitoring of civil structures are mainly used for modal identification and damage detection. Therefore when modal identification analysis is carried out the quality in the identification of the modes will highly influence the damage detection results. It is also widely recognized that the usefulness of the measured data used for modal identification and damage detection is significantly influenced by the number and locations of sensors. The objective of this study is the numerical implementation of two widely known optimum sensor placement methods in beam-like structures

Keywords: optimum sensor placement, structural damage detection, modal identification, beam-like structures.

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Authors: Popy Bora, L. C. Bora, A. Bhattacharya, Sehnaz S. Ahmed

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Microbial bioagents, viz., Pseudomonas fluorescens, Bacillus subtilis, and Trichoderma viride were assessed for their ability to suppress grey blight caused by Pestalotiopsis theae, a major disease of tea crop in Assam. The expression of defense-related phytochemicals due to the application of these bioagents was also evaluated. The individual bioagents, as well as their combinations, were screened for their bioefficacy against P. theae in vitro using nutrient agar (NA) as basal medium. The treatment comprising a combination of the three bioagents, P. fluorescens, B. subtilis, and T. viride showed significantly the highest inhibition against the pathogen. Bioformulation of effective bioagent combinations was further evaluated under field condition, where significantly highest reduction of grey blight (90.30%), as well as the highest increase in the green leaf yield (10.52q/ha), was recorded due to application of the bioformulation containing the three bioagents. The application of the three bioformulation also recorded an enhanced level of caffeine (4.15%) and polyphenols (22.87%). A significant increase in the enzymatic activity of phenylalanine ammonia-lyase, peroxidase and polyphenol oxidase were recorded in the plants treated with the microbial bioformulation of the three bioagents. The present investigation indicates the role of microbial agents in suppressing disease, inducing plant defense response, as well as improving the quality of tea.

Keywords: enzymatic activity, grey blight, microbial bioagents, Pestalotiopsis theae, phytochemicals, plant defense, tea

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Authors: Promil Mehra, Nanthi Bolan, Jack Desbiolles, Risha Gupta

Abstract:

Agricultural practices affect the soil physical and chemical properties, which in turn influence the soil microorganisms as a function of the soil biological environment. On the return to conventional tillage (CT) from continuing no-till (NT) cropping system, a very little information is available from the impact caused by the intermittent tillage on the soil biochemical properties from a short-term (2-year) study period. Therefore, the contribution made by different microorganisms (fungal, bacteria) was also investigated in order to find out the effective changes in the soil microbial activity under a South Australian dryland faring system. This study was conducted to understand the impact of microbial dynamics on the soil organic carbon (SOC) under NT and CT systems when treated with different levels of mulching (0, 2.5 and 5 t/ha). Our results demonstrated that from the incubation experiment the cumulative CO2 emitted from CT system was 34.5% higher than NT system. Relatively, the respiration from surface layer (0-10 cm) was significantly (P<0.05) higher by 8.5% and 15.8 from CT; 8% and 18.9% from NT system w.r.t 10-20 and 20-30 cm respectively. Further, the dehydrogenase enzyme activity (DHA) and microbial biomass carbon (MBC) were both significantly lower (P<0.05) under CT, i.e., 7.4%, 7.2%, 6.0% (DHA) and 19.7%, 15.7%, 4% (MBC) across the different mulching levels (0, 2.5, 5 t/ha) respectively. In general, it was found that from both the tillage system the enzyme activity and MBC decreased with the increase in depth (0-10, 10-20 and 20-30 cm) and with the increase in mulching rate (0, 2.5 and 5 t/ha). From the perspective of microbial stress, there was 28.6% higher stress under CT system compared to NT system. Whereas, the microbial activity of different microorganisms like fungal and bacterial activities were determined by substrate-induced inhibition respiration using antibiotics like cycloheximide (16 mg/gm of soil) and streptomycin sulphate (14 mg/gm of soil), by trapping the CO2 using an alkali (0.5 M NaOH) solution. The microbial activities were confirmed through platting technique, where it was that found bacterial activities were 46.2% and 38.9% higher than fungal activity under CT and NT system. In conclusion, it was expected that changes in the relative abundance and activity of different microorganisms (bacteria and fungi) under different tillage systems could significantly affect the C cycling and storage due to its unique structures and differential interactions with the soil physical properties.

Keywords: tillage, soil respiration, MBC, fungal-bacterial activity

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Authors: Elsayed Ahmed Elsayed, Azza Noor El-Deen, Mohamed A. Farid, Mohamed A. Wadaan

Abstract:

Recently, a great attention has been paid to the use of dietary carbohydrates as prebiotic functional foods. Among the new commercially available products, fructooligosaccharides (FOS), which are microbial produced from sucrose, have attracted special interest due to their valuable properties and, thus, have a great economic potential for the sugar industrial branch. They are non-cariogenic sweeteners of low caloric value, as they are not hydrolyzed by the gastro-intestinal enzymes, promoting selectively the growth of the bifidobacteria in the colon, helping to eliminate the harmful microbial species to human and animal health and preventing colon cancer. FOS has been also found to reduce cholesterol, phospholipids and triglyceride levels in blood. FOS has been mainly produced by microbial fructosyltransferase (FTase) enzymes. The present work outlines bioprocess optimization for different cultivation parameters affecting the production of FTase by Penicillium aurantiogriseum AUMC 5605. The optimization involves both traditional as well as fractional factorial design approaches. Additionally, the production process will be compared under batch and fed-batch conditions. Finally, the optimized process conditions will be applied to 5-L stirred tank bioreactor cultivations.

Keywords: prebiotics, fructooligosaccharides, optimization, cultivation

Procedia PDF Downloads 387

Authors: Kanchan Vishwakarma, Shivesh Sharma, Nitin Kumar

Abstract:

Soil is incoherent matter on Earth’s surface having organic and mineral content. The spatial variation of 4 soil enzyme activities and microbial biomass were assessed for two seasons’ viz. monsoon and winter along the latitudinal gradient in North-central India as the area of this study is fettered with respect to national status. The study was facilitated to encompass the effect of climate change, enzyme activity and biomass on nutrient cycling. Top soils were sampled from 4 sites in North-India. There were significant correlations found between organic C, N & P wrt to latitude gradient in two seasons. This distribution of enzyme activities and microbial biomass was consequence of alterations in temperature and moisture of soil because of which soil properties change along the latitude transect.

Keywords: latitude gradient, microbial biomass, moisture, soil, organic carbon, temperature

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Authors: Mathias Marquardt, Peter Dünow, Sandra Baßler

Abstract:

The paper describes the use of subspace based identification methods for auto tuning of a state space control system. The plant is an unstable but self balancing transport robot. Because of the unstable character of the process it has to be identified from closed loop input-output data. Based on the identified model a state space controller combined with an observer is calculated. The subspace identification algorithm and the controller design procedure is combined to a auto tuning method. The capability of the approach was verified in a simulation experiments under different process conditions.

Keywords: auto tuning, balanced robot, closed loop identification, subspace identification

Procedia PDF Downloads 380

Authors: M. S. Matlala, I. Ignatious

Abstract:

Environmental sustainability has taken the center stage in human life all over the world. Energy is the most essential component of our life. The conventional sources of energy are non-renewable and have a detrimental environmental impact. Therefore, there is a need to move from conventional to non-conventional renewable energy sources to satisfy the world’s energy demands. The study aimed at screening for microbial cellulolytic enzymes using a proteomic approach. The objectives were to screen for microbial cellulases with high specific activity and separate the cellulolytic enzymes using a combination of zymography and two-dimensional (2-D) gel electrophoresis followed by tryptic digestion, Matrix-assisted Laser Desorption Ionisation-Time of Flight (MALDI-TOF) and bioinformatics analysis. Fungal and bacterial isolates were cultured in M9 minimal and Mandel media for a period of 168 hours at 60°C and 30°C with cellobiose and Avicel as carbon sources. Microbial cells were separated from supernatants through centrifugation, and the crude enzyme from the cultures was used for the determination of cellulase activity, zymography, SDS-PAGE, and two-dimensional gel electrophoresis. Five isolates, with lytic action on carbon sources studied, were a bacterial strain (BARK) and fungal strains (VCFF1, VCFF14, VCFF17, and VCFF18). Peak cellulase production by the selected isolates was found to be 3.8U/ml, 2.09U/ml, 3.38U/ml, 3.18U/ml, and 1.95U/ml, respectively. Two-dimensional gel protein maps resulted in the separation and quantitative expression of different proteins by the microbial isolates. MALDI-TOF analysis and database search showed that the expressed proteins in this study closely relate to different glycoside hydrolases produced by other microbial species with an acceptable confidence level of 100%.

Keywords: cellulases, energy, two-dimensional gel electrophoresis, matrix-assisted laser desorption ionisation-time of flight, MALDI-TOF MS

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Authors: Tak-Chung Fu, Ying-Kit Hung, Fu-Lai Chung

Abstract:

In the field of time series data mining, the concept of the Perceptually Important Point (PIP) identification process is first introduced in 2001. This process originally works for financial time series pattern matching and it is then found suitable for time series dimensionality reduction and representation. Its strength is on preserving the overall shape of the time series by identifying the salient points in it. With the rise of Big Data, time series data contributes a major proportion, especially on the data which generates by sensors in the Internet of Things (IoT) environment. According to the nature of PIP identification and the successful cases, it is worth to further explore the opportunity to apply PIP in time series ‘Big Data’. However, the performance of PIP identification is always considered as the limitation when dealing with ‘Big’ time series data. In this paper, two distributed versions of PIP identification based on the Specialized Binary (SB) Tree are proposed. The proposed approaches solve the bottleneck when running the PIP identification process in a standalone computer. Improvement in term of speed is obtained by the distributed versions.

Keywords: distributed computing, performance analysis, Perceptually Important Point identification, time series data mining

Procedia PDF Downloads 433