Search results for: ammonia oxidizing bacteria

Authors: Jesús M. Martín, Francisco Corona, Dolores Hidalgo

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Landfills present long-term threats to soil, air, groundwater and surface water due to the formation of greenhouse gases (methane gas and carbon dioxide) and leachate from decomposing garbage. The composition of leachate differs from site to site and also within the landfill. The leachates alter with time (from weeks to years) since the landfilled waste is biologically highly active and their composition varies. Mainly, the composition of the leachate depends on factors such as characteristics of the waste, the moisture content, climatic conditions, degree of compaction and the age of the landfill. Therefore, the leachate composition cannot be generalized and the traditional treatment models should be adapted in each case. Although leachate composition is highly variable, what different leachates have in common is hazardous constituents and their potential eco-toxicological effects on human health and on terrestrial ecosystems. Since leachate has distinct compositions, each landfill or dumping site would represent a different type of risk on its environment. Nevertheless, leachates consist always of high organic concentration, conductivity, heavy metals and ammonia nitrogen. Leachate could affect the current and future quality of water bodies due to uncontrolled infiltrations. Therefore, control and treatment of leachate is one of the biggest issues in urban solid waste treatment plants and landfills design and management. This work presents a treatment model that will be carried out "in-situ" using a cost-effective novel technology that combines solar evaporation/condensation plus forward osmosis. The plant is powered by renewable energies (solar energy, biomass and residual heat), which will minimize the carbon footprint of the process. The final effluent quality is very high, allowing reuse (preferred) or discharge into watercourses. In the particular case of this work, the final effluents will be reused for cleaning and gardening purposes. A minority semi-solid residual stream is also generated in the process. Due to its special composition (rich in metals and inorganic elements), this stream will be valorized in ceramic industries to improve the final products characteristics.

Keywords: forward osmosis, landfills, leachate valorization, solar evaporation

Procedia PDF Downloads 198

Authors: Piyush Sharma, Devi Prasad Sharma, Sulabh Bansal

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Diseases are fairly common in fruits and vegetables because of the changing climatic and environmental circumstances. Crop diseases, which are frequently difficult to control, interfere with the growth and output of the crops. Accurate disease detection and timely disease control measures are required to guarantee high production standards and good quality. In India, apples are a common crop that may be afflicted by a variety of diseases on the fruit, stem, and leaves. It is fungi, bacteria, and viruses that trigger the early symptoms of leaf diseases. In order to assist farmers and take the appropriate action, it is important to develop an automated system that can be used to detect the type of illnesses. Machine learning-based image processing can be used to: this research suggested a system that can automatically identify diseases in apple fruit and apple plants. Hence, this research utilizes the hybrid SVM-RBN model. As a consequence, the model may produce results that are more effective in terms of accuracy, precision, recall, and F1 Score, with respective values of 96%, 99%, 94%, and 93%.

Keywords: fruit plant disease, crop disease, machine learning, image processing, SVM-RBN

Procedia PDF Downloads 55

Authors: Mahdi Asghari Ozma

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Background and Objective:Maclurapomifera (Rafin.) Schneider, known as osage orange, is a north american native plant which has multiple applications in herbal medicine. The extract of this plant has many therapeutic effects, including antimicrobial, anti-tumor, anti-inflammation, etc., that discussed in this study. Materials and Methods: For this study, the keywords "Maclurapomifera", "osage orange, ""herbal medicine ", and "plant extract" in the databases PubMed and Google Scholar between 2002 and 2021 were searched, and 20 articles were chosen, studied and analyzed. Results: Due to the increased resistance of microbes to antibiotics, the need for antimicrobial plants is increasing. Maclurapomifera is one of the plants with antimicrobial properties that can affect all microbes, especially Gram-negative bacteria, and fungi. This plant also has anti-tumor, anti-inflammatory, anti-oxidant, anti-aging, antiviral, anti-fungal, anti-ulcerogenic, anti-diabetic, and anti-nociceptive effects, which can be used as a substance with many amazing therapeutic applications. Conclusion: These results suggest that the extract of Maclurapomifera can be used in clinical medicine as a remedial agent, which can be substituted for chemical drugs or help them in the treatment of diseases.

Keywords: maclura pomifera, osage orange, herbal medicine, plant extract

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Authors: Palash Soni, Vivek Kumar Gaba, Shubhankar Bhowmick, Bidyut Mazumdar

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Refrigeration technology is a fast developing field at the present era since it has very wide application in both domestic and industrial areas. It started from the usage of simple ice coolers to store food stuffs to the present sophisticated cold storages along with other air conditioning system. A variety of techniques are used to bring down the temperature below the ambient. Adsorption refrigeration technology is a novel, advanced and promising technique developed in the past few decades. It gained attention due to its attractive property of exploiting unlimited natural sources like solar energy, geothermal energy or even waste heat recovery from plants or from the exhaust of locomotives to fulfill its energy need. This will reduce the exploitation of non-renewable resources and hence reduce pollution too. This work is aimed to develop a model for a solar adsorption refrigeration system and to simulate the same for different operating conditions. In this system, the mechanical compressor is replaced by a thermal compressor. The thermal compressor uses renewable energy such as solar energy and geothermal energy which makes it useful for those areas where electricity is not available. Refrigerants normally in use like chlorofluorocarbon/perfluorocarbon have harmful effects like ozone depletion and greenhouse warming. It is another advantage of adsorption systems that it can replace these refrigerants with less harmful natural refrigerants like water, methanol, ammonia, etc. Thus the double benefit of reduction in energy consumption and pollution can be achieved. A thermodynamic model was developed for the proposed adsorber, and a universal MATLAB code was used to simulate the model. Simulations were carried out for a different operating condition for the silicagel-methanol working pair. Various graphs are plotted between regeneration temperature, adsorption capacities, the coefficient of performance, desorption rate, specific cooling power, adsorption/desorption times and mass. The results proved that adsorption system could be installed successfully for refrigeration purpose as it has saving in terms of power and reduction in carbon emission even though the efficiency is comparatively less as compared to conventional systems. The model was tested for its compliance in a cold storage refrigeration with a cooling load of 12 TR.

Keywords: adsorption, refrigeration, renewable energy, silicagel-methanol

Procedia PDF Downloads 201

Authors: Arifur Rahman, Ardeshir Amirkhani, Honghua Hu, Mark Molloy, Karen Vickery

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Introduction and Objectives: Staphylococcus aureus and coagulase-negative staphylococci comprises approximately 65% of infections associated with medical devices and are well known for their biofilm formatting ability. Biofilm-related infections are extremely difficult to eradicate owing to their high tolerance to antibiotics and host immune defences. Currently, there is no efficient method for early biofilm detection. A better understanding to enable detection of biofilm specific proteins in vitro and in vivo can be achieved by studying planktonic and different growth phases of biofilms using a proteome analysis approach. Our goal was to construct a reference map of planktonic and biofilm associated proteins of S. aureus. Methods: S. aureus reference strain (ATCC 25923) was used to grow 24 hours planktonic, 3-day wet biofilm (3DWB), and 12-day wet biofilm (12DWB). Bacteria were grown in tryptic soy broth (TSB) liquid medium. Planktonic growth was used late logarithmic bacteria, and the Centres for Disease Control (CDC) biofilm reactor was used to grow 3 days, and 12-day hydrated biofilms, respectively. Samples were subjected to reduction, alkylation and digestion steps prior to Multiplex labelling using Tandem Mass Tag (TMT) 10-plex reagent (Thermo Fisher Scientific). The labelled samples were pooled and fractionated by high pH RP-HPLC which followed by loading of the fractions on a nanoflow UPLC system (Eksigent UPLC system, AB SCIEX). Mass spectrometry (MS) data were collected on an Orbitrap Elite (Thermo Fisher Scientific) Mass Spectrometer. Protein identification and relative quantitation of protein levels were performed using Proteome Discoverer (version 1.3, Thermo Fisher Scientific). After the extraction of protein ratios with Proteome Discoverer, additional processing, and statistical analysis was done using the TMTPrePro R package. Results and Discussion: The present study showed that a considerable proteomic difference exists among planktonic and biofilms from S. aureus. We identified 1636 total extracellular secreted proteins, of which 350 and 137 proteins of 3DWB and 12DWB showed significant abundance variation from planktonic preparation, respectively. Of these, simultaneous up-regulation in between 3DWB and 12DWB proteins such as extracellular matrix-binding protein ebh, enolase, transketolase, triosephosphate isomerase, chaperonin, peptidase, pyruvate kinase, hydrolase, aminotransferase, ribosomal protein, acetyl-CoA acetyltransferase, DNA gyrase subunit A, glycine glycyltransferase and others we found in this biofilm producer. On the contrary, simultaneous down-regulation in between 3DWB and 12DWB proteins such as alpha and delta-hemolysin, lipoteichoic acid synthase, enterotoxin I, serine protease, lipase, clumping factor B, regulatory protein Spx, phosphoglucomutase, and others also we found in this biofilm producer. In addition, we also identified a big percentage of hypothetical proteins including unique proteins. Therefore, a comprehensive knowledge of planktonic and biofilm associated proteins identified by S. aureus will provide a basis for future studies on the development of vaccines and diagnostic biomarkers. Conclusions: In this study, we constructed an initial reference map of planktonic and various growth phase of biofilm associated proteins which might be helpful to diagnose biofilm associated infections.

Keywords: bacterial biofilms, CDC bioreactor, S. aureus, mass spectrometry, TMT

Procedia PDF Downloads 164

Authors: Shagufta Jabeen, Faez J. Firdaus Abdullah, Zunita Zakaria, Nurulfiza M. Isa, Yung C. Tan, Wai Y. Yee, Abdul R. Omar

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Pasteurella multocida (PM) is an important veterinary opportunistic pathogen particularly associated with septicemic pasteurellosis, pneumonic pasteurellosis and hemorrhagic septicemia in cattle and buffaloes. P. multocida serotype A has been reported to cause fatal pneumonia and septicemia. Pasteurella multocida subspecies multocida of serotype A Malaysian isolate PMTB2.1 was first isolated from buffaloes died of septicemia. In this study, the genome of P. multocida strain PMTB2.1 was sequenced using third-generation sequencing technology, PacBio RS2 system and analyzed bioinformatically via de novo analysis followed by in-depth analysis based on comparative genomics. Bioinformatics analysis based on de novo assembly of PacBio raw reads generated 3 contigs followed by gap filling of aligned contigs with PCR sequencing, generated a single contiguous circular chromosome with a genomic size of 2,315,138 bp and a GC content of approximately 40.32% (Accession number CP007205). The PMTB2.1 genome comprised of 2,176 protein-coding sequences, 6 rRNA operons and 56 tRNA and 4 ncRNAs sequences. The comparative genome sequence analysis of PMTB2.1 with nine complete genomes which include Actinobacillus pleuropneumoniae, Haemophilus parasuis, Escherichia coli and five P. multocida complete genome sequences including, PM70, PM36950, PMHN06, PM3480, PMHB01 and PMTB2.1 was carried out based on OrthoMCL analysis and Venn diagram. The analysis showed that 282 CDs (13%) are unique to PMTB2.1and 1,125 CDs with orthologs in all. This reflects overall close relationship of these bacteria and supports the classification in the Gamma subdivision of the Proteobacteria. In addition, genomic distance analysis among all nine genomes indicated that PMTB2.1 is closely related with other five Pasteurella species with genomic distance less than 0.13. Synteny analysis shows subtle differences in genetic structures among different P.multocida indicating the dynamics of frequent gene transfer events among different P. multocida strains. However, PM3480 and PM70 exhibited exceptionally large structural variation since they were swine and chicken isolates. Furthermore, genomic structure of PMTB2.1 is more resembling that of PM36950 with a genomic size difference of approximately 34,380 kb (smaller than PM36950) and strain-specific Integrative and Conjugative Elements (ICE) which was found only in PM36950 is absent in PMTB2.1. Meanwhile, two intact prophages sequences of approximately 62 kb were found to be present only in PMTB2.1. One of phage is similar to transposable phage SfMu. The phylogenomic tree was constructed and rooted with E. coli, A. pleuropneumoniae and H. parasuis based on OrthoMCL analysis. The genomes of P. multocida strain PMTB2.1 were clustered with bovine isolates of P. multocida strain PM36950 and PMHB01 and were separated from avian isolate PM70 and swine isolates PM3480 and PMHN06 and are distant from Actinobacillus and Haemophilus. Previous studies based on Single Nucleotide Polymorphism (SNPs) and Multilocus Sequence Typing (MLST) unable to show a clear phylogenetic relatedness between Pasteurella multocida and the different host. In conclusion, this study has provided insight on the genomic structure of PMTB2.1 in terms of potential genes that can function as virulence factors for future study in elucidating the mechanisms behind the ability of the bacteria in causing diseases in susceptible animals.

Keywords: comparative genomics, DNA sequencing, phage, phylogenomics

Procedia PDF Downloads 182

Authors: F. Z. Aliyat, B. Ben Messaoud, L. Nassiri, E. Bouiamrine, J. Ibijbijen

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The present study aims to assess the biological nitrogen fixation in the soybean tested in different Moroccan soils combined with the rhizobial inoculation. These effects were evaluated by the plant growth mainly by the aerial biomass production, total nitrogen content and the proportion of the nitrogen fixed. This assessment clearly shows that the inoculation with bacteria increases the growth of soybean. Five different soils and a control (peat) were used. The rhizobial inoculation was performed by applying the peat that contained a mixture of 2 strains Sinorhizobium fredii HH103 and Bradyrhizobium. The biomass, the total nitrogen content and the proportion of nitrogen fixed were evaluated under different treatments. The essay was realized at the greenhouse the Faculty of Sciences, Moulay Ismail University. The soybean has shown a great response for the parameters assessed. Moreover, the best response was reported by the inoculated plants compared to non- inoculated and to the absolute control. Finally, good production and the best biological nitrogen fixation present an important ecological technology to improve the sustainable production of soybean and to ensure the increase of the fertility of soils.

Keywords: biological nitrogen fixation, inoculation, rhizobium, soybean

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Authors: Aashutosh Garg, Ankur Rajpal, A. A. Kazmi

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The efficiency of an on-site wastewater treatment system named Johkasou was evaluated based on its pollutant removal efficiency over 10 months. This system was installed at IIT Roorkee and had a capacity of treating 7 m3/d of sewage water, sufficient for a group of 30-50 people. This system was fed with actual wastewater through an equalization tank to eliminate the fluctuations throughout the day. Methanol and ammonium chloride was added into this equalization tank to increase the Chemical Oxygen Demand (COD) and ammonia content of the influent. The outlet from Johkasou is sent to a tertiary unit consisting of a Pressure Sand Filter and an Activated Carbon Filter for further treatment. Samples were collected on alternate days from Monday to Friday and the following parameters were evaluated: Chemical Oxygen Demand (COD), Biochemical Oxygen Demand (BOD), Total Suspended Solids (TSS), and Total Nitrogen (TN). The Average removal efficiency for Chemical Oxygen Demand (COD), Biochemical Oxygen Demand (BOD), Total Suspended Solids (TSS), and Total Nitrogen (TN) was observed as 89.6, 97.7, 96, and 80% respectively. The cost of treating the wastewater comes out to be Rs 23/m3 which includes electricity, cleaning and maintenance, chemical, and desludging costs. Tests for the coliforms were also performed and it was observed that the removal efficiency for total and fecal coliforms was 100%. The sludge generation rate is approximately 20% of the BOD removal and it needed to be removed twice a year. It also showed a very good response against the hydraulic shock load. We performed vacation stress analysis on the system to evaluate the performance of the system when there is no influent for 8 consecutive days. From the result of stress analysis, we concluded that system needs a recovery time of about 48 hours to stabilize. After about 2 days, the system returns again to original conditions and all the parameters in the effluent become within the limits of National Green Tribunal (NGT) standards. We also performed another stress analysis to save the electricity in which we turned the main aeration blower off for 2 to 12 hrs a day and the results showed that we can turn the blower off for about 4-6 hrs a day and this will help in reducing the electricity costs by about 25%. It was concluded that the Johkasou system can remove a sufficient amount of all the physiochemical parameters tested to satisfy the prescribed limit set as per Indian Standard.

Keywords: on-site treatment, domestic wastewater, Johkasou, nutrient removal, pathogens removal

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Authors: Serdar Yaman, Hanzade Haykiri-Acma

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Biomass is a CO₂-neutral fuel that is renewable and sustainable along with having very huge global potential. Efficient use of biomass in power generation and production of biomass-based biofuels can mitigate the greenhouse gasses (GHG) and reduce dependency on fossil fuels. There are also other beneficial effects of biomass energy use such as employment creation and pollutant reduction. However, most of the biomass materials are not capable of competing with fossil fuels in terms of energy content. High moisture content and high volatile matter yields of biomass make it low calorific fuel, and it is very significant concern over fossil fuels. Besides, the density of biomass is generally low, and it brings difficulty in transportation and storage. These negative aspects of biomass can be overcome by thermal pretreatments that upgrade the fuel property of biomass. That is, torrefaction is such a thermal process in which biomass is heated up to 300ºC under non-oxidizing conditions to avoid burning of the material. The treated biomass is called as biochar that has considerably lower contents of moisture, volatile matter, and oxygen compared to the parent biomass. Accordingly, carbon content and the calorific value of biochar increase to the level which is comparable with that of coal. Moreover, hydrophilic nature of untreated biomass that leads decay in the structure is mostly eliminated, and the surface properties of biochar turn into hydrophobic character upon torrefaction. In order to investigate the effectiveness of torrefaction process on biomass properties, several biomass species such as olive milling residue (OMR), Rhododendron (small shrubby tree with bell-shaped flowers), and ash tree (timber tree) were chosen. The fuel properties of these biomasses were analyzed through proximate and ultimate analyses as well as higher heating value (HHV) determination. For this, samples were first chopped and ground to a particle size lower than 250 µm. Then, samples were subjected to torrefaction in a horizontal tube furnace by heating from ambient up to temperatures of 200, 250, and 300ºC at a heating rate of 10ºC/min. The biochars obtained from this process were also tested by the methods applied to the parent biomass species. Improvement in the fuel properties was interpreted. That is, increasing torrefaction temperature led to regular increases in the HHV in OMR, and the highest HHV (6065 kcal/kg) was gained at 300ºC. Whereas, torrefaction at 250ºC was seen optimum for Rhododendron and ash tree since torrefaction at 300ºC had a detrimental effect on HHV. On the other hand, the increase in carbon contents and reduction in oxygen contents were determined. Burning characteristics of the biochars were also studied using thermal analysis technique. For this purpose, TA Instruments SDT Q600 model thermal analyzer was used and the thermogravimetric analysis (TGA), derivative thermogravimetry (DTG), differential scanning calorimetry (DSC), and differential thermal analysis (DTA) curves were compared and interpreted. It was concluded that torrefaction is an efficient method to upgrade the fuel properties of biomass and the biochars from which have superior characteristics compared to the parent biomasses.

Keywords: biochar, biomass, fuel upgrade, torrefaction

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Authors: Mayoorika Shukla, Pramila Jakhar, Tejendra Dixit, I. A. Palani, Vipul Singh

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Biosensors are analytical devices with wide range of applications in biological, chemical, environmental and clinical analysis. It comprises of bio-recognition layer which has biomolecules (enzymes, antibodies, DNA, etc.) immobilized over it for detection of analyte and transducer which converts the biological signal into the electrical signal. The performance of biosensor primarily the depends on the bio-recognition layer and therefore it has to be chosen wisely. In this regard, nanostructures of metal oxides such as ZnO, SnO2, V2O5, and TiO2, etc. have been explored extensively as bio-recognition layer. Recently, ZnO has the attracted attention of researchers due to its unique properties like high iso-electric point, biocompatibility, stability, high electron mobility and high electron binding energy, etc. Although there have been many reports on usage of ZnO as bio-recognition layer but to the authors’ knowledge, none has ever observed correlation between optical properties like defect suppression and biosensing capability of the sensor. Here, ZnO nanorods (ZNR) have been synthesized by a low cost, simple and low-temperature hydrothermal growth process, over Platinum (Pt) coated glass substrate. The ZNR have been synthesized in two steps viz. initially a seed layer was coated over substrate (Pt coated glass) followed by immersion of it into nutrient solution of Zinc nitrate and Hexamethylenetetramine (HMTA) with in situ addition of KMnO4. The addition of KMnO4 was observed to have a profound effect over the growth rate anisotropy of ZnO nanostructures. Clustered and powdery growth of ZnO was observed without addition of KMnO4, although by addition of it during the growth, uniform and crystalline ZNR were found to be grown over the substrate. Moreover, the same has resulted in suppression of defects as observed by Normalized Photoluminescence (PL) spectra since KMnO4 is a strong oxidizing agent which provides an oxygen rich growth environment. Further, to explore the correlation between defect suppression and biosensing capability of the ZNR Glucose oxidase (Gox) was immobilized over it, using physical adsorption technique followed by drop casting of nafion. Here the main objective of the work was to analyze effect of defect suppression over biosensing capability, and therefore Gox has been chosen as model enzyme, and electrochemical amperometric glucose detection was performed. The incorporation of KMnO4 during growth has resulted in variation of optical and charge transfer properties of ZNR which in turn were observed to have deep impact on biosensor figure of merits. The sensitivity of biosensor was found to increase by 12-18 times, due to variations introduced by addition of KMnO4 during growth. The amperometric detection of glucose in continuously stirred buffer solution was performed. Interestingly, defect suppression has been observed to contribute towards the improvement of biosensor performance. The detailed mechanism of growth of ZNR along with the overall influence of defect suppression on the sensing capabilities of the resulting enzymatic electrochemical biosensor and different figure of merits of the biosensor (Glass/Pt/ZNR/Gox/Nafion) will be discussed during the conference.

Keywords: biosensors, defects, KMnO4, ZnO nanorods

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Authors: Frank Boris Feutmba Keutchou, Saurelle Fabienne Bieghan Same, Verelle Elsa Fogang Pokam, Charles Ursula Metapi Meikeu, Angel Marilyne Messop Nzomo, Ousman Tamgue

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Infectious diseases are one of the most important causes of morbidity and mortality worldwide. These diseases are caused by micro-pathogenic organisms, such as bacteria, viruses, parasites, and fungi. Heritable changes in gene expression that do not involve changes to the underlying DNA sequence are referred to as epigenetics. Emerging evidence suggests that epigenetic mechanisms are important in the emergence and progression of infectious diseases. Pathogens can manipulate host epigenetic machinery to promote their own replication and evade immune responses. The Human Genome Project has provided new opportunities for developing better tools for the diagnosis and identification of target genes. Several epigenetic modifications, such as DNA methylation, histone modifications, and non-coding RNA expression, have been shown to influence infectious disease outcomes. Understanding the epigenetic mechanisms underlying infectious diseases may result in the progression of new therapeutic approaches focusing on host-pathogen interactions. The goal of this study is to show how different infectious agents interact with host cells after infection.

Keywords: epigenetic, infectious disease, micro-pathogenic organism, phenotype

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Authors: Muhammad Arslan, Michael A. Kaminski, Bassam S. Tawabini, Fabrizio Frontalini

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We conducted a survey of living benthic foraminifera in a relatively unpolluted site of Bahrain in the Arabian Gulf, with the aim of determining the seasonal variability in their populations, as well as various environmental parameters that affect their distribution. The maximum standing crop was observed during winter, with highest population of rotaliids, followed by a peak in miliolids. The highest population is attributed to an increasing number juveniles observed along the depth transect. A strong correlation between sediment grain size and the foraminiferal population indicates that juveniles were most abundant on coarser sandy substrate and less abundant on fine substrate. In spring, the total living population decreased, and lowest values are observed in the summer. The population started to increase again in the autumn with highest juveniles/adult ratios. Moreover, results of relative abundance and species consistency show that Ammonia is found to be consistent from the shallowest to the deepest station, whereas miliolids start appearing in the deeper stations. The average numbers of Peneroplis and Elphidium also increases along the depth transect. Environmental characterization reveals that although the site is subjected to eutrophication caused by nitrates and sulfates, pollution caused by hydrocarbons and heavy metals is not significant. The assessment of 63 heavy metals showed that none of the metals had concentrations that exceed internationally accepted norms [the devised level of Effect Range-Low], with the exception of strontium. The lack of a significant environmental effect of heavy metals is confirmed by a Foraminiferal Deformities Index value of less than 2%. Likewise, no hydrocarbon contamination was detected in the water or sediment samples. Lastly, observations of cytoplasmic streaming and pseudopodial activity in Petri dishes suggest that the foraminiferal population is not stressed. We conclude that the site in Bahrain is not yet adversely affected by human development, and therefore can provide baseline information for future comparison and assessment of foraminiferal assemblages in contaminated zones of the Arabian Gulf.

Keywords: Arabian Gulf, benthic foraminifera, standing crop, Western Bahrain

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Authors: Haruna Saidu, Sulaiman Mohammed, Abdulkarim Ali Deba, Shaza Eva Mohamad

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Determining the diversity of the indigenous microorganisms in Palm Oil Mill Effluent (POME) could allow their wider application for the treatment of recalcitrant agro-based wastewater discharge into the environment. Many research studies mainly determined the efficiency of microorganism or their co-cultivation with microalgae for enhanced treatment of wastewater, suggesting a limited emphasis on the application of microbial diversity. In this study, the microorganism was cultured in POME for a period of 15 days using microalgae as a source of carbon. Pyrosequencing analysis reveals a diversity of microbial community in 20% (v/v) culture than the control experiment. Most of the bacterial species identified in POME belong to the families of Bacillaceae, Paenibacillaceae, Enterococcaceae, Clostridiaceae, Peptostreptococcaceae, Caulobacteraceae, Enterobacteriaceae, Moraxellaceae, and Pseudomonadaceae. Alpha (α) diversity analysis reveals the high composition of the microbial community of 52 in both samples. Beta (β) diversity index indicated the occurrence of similar species of microorganisms in unweighted uni fra than the weighted uni fra of both samples. It is therefore suggested that bacteria found in these families could have a potential for synergistic treatment of high-strength wastewater generated from the palm oil industry.

Keywords: diversity, microorganism, wastewater, pyrosequencing, palm oil mill effluent

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Authors: Boudjelal Amel

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Arisarum vulgare, a member of the Araceae family, is an herbaceous perennial widely distributed in the Mediterranean region. A. vulgare is recognized for its medicinal properties and holds significant traditional importance in Algeria for the treatment of various human ailments, including pain, infections, inflammation, digestive disorders, skin problems, eczema, cancer, wounds, burns and gynecological diseases. Despite its extensive traditional use, scientific exploration of A. vulgare remains limited. The study aims to investigate for the first time the therapeutic potential of A. vulgare ethanolic extract obtained by ultrasound-assisted extraction. The chemical composition of the extract was determined by LC-MS/MS analysis. For in vitro phytopharmacological evaluation, several assays, including DPPH, ABTS, FRAP and reducing power, were employed to evaluate the antioxidant activity. The antibacterial activity was assessed againt Escherichia coli, Salmonella typhimurium, Staphylococus aureus, Enterococcus feacium by disk diffusion and microdilution methods. The possible inhibitory activity of ethanolic extract was analyzed against the cholinesterases enzymes (AChE and BChE). The DNA protection activity of A. vulgare ethanolic extract was estimated using the agarose gel electrophoresis method. The capacities of the extract to protect plasmid DNA (pBR322) from the oxidizing effects of H2O2 and UV treatment were evaluated by their DNA-breaking forms. The in vivo wound healing potential of a traditional ointment containing 5% of A. vulgare ethanolic extract was also investigated. The LC-MS/MS profiling of the extract revealed the presence of various bioactive compounds, including naringenin, chlorogenic, vanillic, cafeic, coumaric acids, trans-cinnamic and trans ferrulic acids. The plant extract presented considerable antioxidant potential, being the most active for Reducing power (0,07326±0.001 mg/ml) and DPPH (0.14±0.004 mg/ml). The extract showed the highest inhibition zone diameter against Enterococcus feacium (36±0.1 mm). The ethanolic extract of A. vulgare suppressed the growth of Staphylococus aureus, Escherichia coli and Salmonella typhimurium according to the MIC values. The extract of the plant significantly inhibited both AChE and BChE enzymes. DNA protection activity of the A. vulgare extract was determined as 90.41% for form I and 51.92% for form II. The in vivo experiments showed that 5% ethanolic extract ointment accelerated the wound healing process. The topical application of the traditional formulation enhanced wound closure (95,36±0,6 %) and improved histological parameters in the treated group compared to the control groups. The promising biological properties of Arisarum vulgare revealed that the plant could be appraised as a potential origin of bioactive molecules having multifunctional medicinal uses.

Keywords: arisarum vulgare, LC-MS/MS, antioxidant activity, antimicrobial activity, cholinesterases enzymes inhibition, dna-damage activity, in vivo wound healing

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Authors: Kinza Khan, Dad Muhmmad, Asif Saleem, Nadia Mukhtar, Tahir Yaqub

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Ethomedicines are more commonly used in underdeveloped and developing countries. These medicines are sometimes more potent in controlling microbial infections than conventional medicines. Medicinal plants have been common practice to cure many diseases for centuries. Murraya koenigii is one of these plants and is commonly used in South Asian countries as a flavoring agent in food. To evaluate its anti-microbial activity, six different bacterial strains (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhi, Bacillus cereus and Klebsiella pneumonia were used. N-hexane extract of Murraya koenigii leaves shows maximum activity against Bacillus cereus. Acetone extract of Murraya koenigii shoots showed more efficient activity against Pseudomonas aeruginosa Dichloromethane extracts showed maximum activity against Bacillus cereus. Ethanol extract exhibited maximum activity against Pseudomonas aeruginosa and Klebsiella pneumoniae. The methanol extract of Murraya koenigii shoots displayed maximum antibacterial activity against Bacillus cereus. Antifungal activity Ethanol extract was more effective against Candida albicans.

Keywords: ethnomedicines, bacteria, fungi, murraya koenigii, antimicrobial activity

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Authors: Mahnoor Khan, Bashir Ahmad, Khizar Hayat, Saad Ahmad Khan, Laiba Ahmad, Shumaila Bashir, Abid Ali Khan

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The objective of this study was to synthesize the smallest possible size of ZnO NPs using a modified wet chemical synthesis method and to prepare core shell using polyethylene glycol (PEG) as shell material. Advanced and sophisticated techniques were used to confirm the synthesis, size, and shape of these NPs. Rounded, clustered NPs of size 5.343 nm were formed. Both the plain and core shell NPs were tested against MDR bacteria (E. cloacae, E. amnigenus, Shigella, S. odorifacae, Citrobacter, and E. coli). Both of the NPs showed excellent antibacterial properties, whereas E. cloacae showed maximum zone of inhibition of 16 mm, 27 mm, and 32 mm for 500 μg/ml, 1000 μg/ml, and 1500 μg/ml, respectively for plain ZnO NPs and 18 mm, 28 mm and 35 mm for 500 μg/ml, 1000 μg/ml and 1500 μg/ml for core shell NPs. These NPs were also biocompatible on human red blood cells showing little hemolysis of only 4% for 70 μg/ml for plain NPs and 1.5% for 70 μg/ml for core shell NPs. Core shell NPs were highly biocompatible because of the PEG. Their therapeutic effect as photosensitizers in photodynamic therapy (PDT) for cancer treatment was also monitored. The cytotoxicity of ZnO and PEG-ZnO was evaluated using MTT assay. Our results demonstrated that these NPs could generate ROS inside tumor cells after irradiation which in turn initiates an apoptotic pathway leading to cell death hence proving to be an effective candidate for PDT.

Keywords: ZnO, hemolysis, cytotoxiciy assay, photodynamic therapy, antibacterial

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Authors: Riccardo Zanni, Maria Galvez-Llompart, Ramon Garcia-Domenech, Jorge Galvez

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Biopesticides, such as naturally occurring chemicals, pheromones, fungi, bacteria and insect predators are often a winning choice in crop protection because of their environmental friendly profile. They are considered to have lower toxicity than traditional pesticides. After almost a century of pesticides use, resistances to traditional insecticides are wide spread, while those to bioinsecticides have raised less attention, and resistance management is frequently neglected. This seems to be a crucial mistake since resistances have already occurred for many marketed biopesticides. With an eye to the future, we present here a selection of new natural occurring chemicals as potential bioinsecticides. The molecules were selected using a consolidated mathematical paradigm called molecular topology. Several QSAR equations were depicted and subsequently applied for the virtual screening of hundred thousands molecules of natural origin, which resulted in the selection of new potential bioinsecticides. The most innovative aspect of this work does not only reside in the importance of the identification of new molecules overcoming biopesticides’ resistances, but on the possibility to promote shared knowledge in the field of green chemistry through this unique in silico discipline named molecular topology.

Keywords: green chemistry, QSAR, molecular topology, biopesticide

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Authors: Mehani Mouna, Boukhari Nadjet, Ladjal Segni

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The aim of our study is to determine the antimicrobial effect of essential oils of two plants Cotula cinerea and Chamomilla recutita on some pathogenic bacteria. It is a medicinal plant used in traditional therapy. Essential oils have many therapeutic properties. In herbal medicine, they are used for their antiseptic properties against infectious diseases of fungal origin, against dermatophytes, those of bacterial origin. Essential oils have many therapeutic properties. In herbal medicine, they are used for their antiseptic properties against infectious diseases of fungal origin, against dermatophytes, those of bacterial origin. Humans use plants for thousands of years to treat various ailments, in many developing countries; much of the population relies on traditional doctors and their collections of medicinal plants to cure them. The test adopted is based on the diffusion method on solid medium (Antibiogram), this method allows to determine the susceptibility or resistance of an organism according to the sample studied. Our study reveals that the essential oil of the plants Cotula cinerea and Chamomilla recutita have a different effect on the resistance of germs.

Keywords: antibiogram, Chamomilla recutita, Cotula cinerea, essential oil, microorganism

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Authors: K. Karapetyan, F. Tkhruni, A. Aghajanyan, T. S. Balabekyan, L. Arstamyan

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Introduction: Globalization of the food supply has created the conditions favorable for the emergence and spread of food-borne and especially dangerous pathogens (EDP) in developing countries. The fresh-cut fruit and vegetable industry is searching for alternatives to replace chemical treatments with biopreservative approaches that ensure the safety of the processed foods product. Antimicrobial compounds of lactic acid bacteria (LAB) possess bactericidal or bacteriostatic activity against intestinal pathogens, spoilage organisms and food-borne pathogens such as Listeria monocytogenes, Staphylococcus aureus and Salmonella. Endemic strains of LAB were isolated. The strains, showing broad spectrum of antimicrobial activity against food spoiling microorganisms, were selected. The genotyping by 16S rRNA sequencing, GS-PCR, RAPD PCR methods showed that they were presented by Lactobacillus rhamnosus109, L.plantarum 65, L.plantarum 66 and Enterococcus faecium 64 species. LAB are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB strains were isolated from different dairy products from rural households from the highland regions of Armenia. Serially diluted samples were spread on MRS (Merck, Germany) and hydrolyzed milk agar (1,2 % w/v). Single colonies from each LAB were individually inoculated in liquid MRS medium and incubated at 37oC for 24 hours. Culture broth with biomass was centrifuged at 10,000 g during 20 min for obtaining of cell free culture broth (CFC). The antimicrobial substances from CFC broth were purified by the combination of adsorption-desorption and ion-exchange chromatography methods. Separation of bacteriocins was performed using a HPLC method on "Avex ODS" C18 column. Mass analysis of peptides recorded on the device API 4000 in the electron ionization mode. The spot-on-lawn method on the test culture plated in the solid medium was applied. The antimicrobial activity is expressed in arbitrary units (AU/ml). Results. Purification of CFC broth of LAB allowed to obtain partially purified antimicrobial preparations which contains bacteriocins with broad spectrum of antimicrobial activity. Investigation of their main biochemical properties shown, that inhibitory activity of preparations is partially reduced after treatment with proteinase K, trypsin, pepsin, suggesting a proteinaceous nature of bacteriocin-like substances containing in CFC broth. Preparations preserved their activity after heat treatment (50-121 oC, 20 min) and were stable in the pH range 3–8. The results of SDS PAAG electrophoresis show that L.plantarum 66 and Ent.faecium 64 strains have one bacteriocin (BCN) with maximal antimicrobial activity with approximate molecular weight 2.0-3.0 kDa. From L.rhamnosus 109 two BCNs were obtained. Mass spectral analysis indicates that these bacteriocins have peptide bonds and molecular weight of BCN 1 and BCN 2 are approximately 1.5 kDa and 700 Da. Discussion: Thus, our experimental data shown, that isolated endemic strains of LAB are able to produce bacteriocins with high and different inhibitory activity against broad spectrum of microorganisms of different taxonomic group, such as Salmonella sp., Esherichia coli, Bacillus sp., L.monocytogenes, Proteus mirabilis, Staph. aureus, Ps. aeruginosa. Obtained results proved the perspectives for use of endemic strains in the preservation of foodstuffs. Acknowledgments: This work was realized with financial support of the Project Global Initiatives for Preliferation Prevention (GIPP) T2- 298, ISTC A-1866.

Keywords: antimicrobial activity, bacteriocins, endemic strains, food safety

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Authors: Hayyan I Al Taweil

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Background: The most common of bacterium in kitchen sponges and cutting surfaces which can play a task within the cross-contamination of foods, fomites and hands by foodborne pathogens. Aims and Objectives: This study investigated the incidence of bacterium in kitchen Sponge, and cutting surfaces. Material and methods: a complete of twenty four kitchen Sponges were collected from home kitchens and therefore the numbers of mesotrophic microorganism, coliform microorganism, E. coli, Salmonella, genus {pseudomonas|bacteria genus} and staphylococci in every kitchen Sponges were determined. Microbiological tests of all sponges for total mesophilic aerobic microorganism, S. aureus, Pseudomonas, Salmonella spp., and E. coli were performed on days 3, 7, and 14 by sampling. The sponges involved in daily use in kitchens countenosely with the dishwasher detergent a minimum of doubly daily Results: Results from the overall mesophilic aerobic microorganism, indicate a major increase within the variety of log CFU/ml. the amount of E. coli was reduced, Salmonella spp. was stabled, S. aureus was enhanced from the sponges throughout fourteen days. Genus Pseudomonas was enhanced and was the dominant micro flora within the sponges throughout fourteen days.

Keywords: Kitchen Sponges, Microbiological Contamination, Disinfection; cutting surface; , Cross-Contamination

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Authors: Hui Zhu, Yuan Wang, Shibo Kuang, Aibing Yu

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The inertial difference induced by the microfluidics inside a curved micro-channel has great potential to provide a fast, inexpensive, and portable solution to the separation of micro- and sub-micro particles in many applications such as aerosol collections, airborne bacteria and virus detections, as well as particle sortation. In this work, the separation behaviors of different sized particles inside a reported curved micro-channel have been studied by a combined approach of computational fluid dynamics for gas and discrete element model for particles (CFD-DEM). The micro-channel is operated by controlling the gas flow rates at all of its branches respectively used to load particles, introduce gas streams, collect particles of various sizes. The validity of the model has been examined by comparing by the calculated separation efficiency of different sized particles against the measurement. On this basis, the separation mechanisms of the inertial microfluidic separator are elucidated in terms of the interactions between particles, between particle and fluid, and between particle and wall. The model is then used to study the effect of feed solids concentration on the separation accuracy and efficiency. The results obtained from the present study demonstrate that the CFD-DEM approach can provide a convenient way to study the particle separation behaviors in micro-channels of various types.

Keywords: CFD-DEM, inertial effect, microchannel, separation

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Authors: Mabtho Moreroa-Monyelo, Grace Ijoma, Rosina Nkuna, Tonderayi Matambo

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A moving bed biofilm reactor (MBBR) was used for the bioremediation of high strength chemical oxygen demand (COD) Fisher-Tropsch (FT) wastewater. The aerobic MBBR system was operated over 60 days. For metagenomics profile assessment of the targeted 16S sequence of bacteria involved in the bioremediation of the chemical compounds, sludge samples were collected every second day of operation. Parameters such as pH and COD were measured daily to compare the system efficiency as the changedin microbial diversity progressed. The study revealed that pH was a contributing factor to microbial diversity, which further affected the efficiency of the MBBR system. The highest COD removal rate of 86.4% was achieved at pH 8.3. It was observed that when there was more, A higher bacterial diversity led to an improvement in the reduction of COD. Furthermore, an OTUof 4530 was obtained, which were divided into 12 phyla, 27 classes, 44 orders, 74 families, and 138 genera across all sludge samples from the MBBR. A determination of the relative abundance of microorganisms at phyla level indicates that the most abundant phylum on day it was Firmicutes (50%); thereafter, the most abundant phylum changed toProteobacteria.

Keywords: biodegradation, fischer-tropsch wastewater, metagenomics, moving bed biofilm reactor

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Authors: Abhipsa Mohanty, Harit Jha

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The world's energy demands are continuing to rise, resulting in a worldwide energy crisis and environmental pollution. Because of finite, declining supply and environmental damage, reliance on fossil fuels is unsustainable. As a result, experts are concentrating on alternative, renewable, and carbon-free energy sources. Energy sources that are both environmentally and economically sustainable are required. Microbial fuel cells (MFCs) have recently received a lot of attention due to their low operating temperatures and ability to use a variety of biodegradable substrates as fuel. There are single-chamber MFCs as well as traditional MFCs with anode and cathode compartments. Bioelectricity is produced when microorganisms actively catabolize substrate. MFCs can be used as a power source in small devices like biosensors. Understanding of its components, microbiological processes, limiting variables, and construction designs in MFC systems must be simplified, and large-scale systems must be developed for them to be cost-effective as well as increase electricity production. The purpose of this research was to review current microbiology knowledge in the field of electricity. The manufacturing process, the materials, and procedures utilized to construct the technology, as well as the applications of MFC technology, are all covered.

Keywords: bio-electricity, exoelectrogenic bacteria, microbial fuel cells, soil microorganisms

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Authors: E. Osazee, L. U. Bashir

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This study was conducted at the Department of Biological Sciences, Usmanu Danfodiyo University, Sokoto, Nigeria, to determine the effects of different weights of cow dung on indigenous bacterial isolates in remediation of crude oil contaminated soils. The soil (1kg) was contaminated with 20g of crude oil and this was treated with three (40g, 80g and 120g) weights of cow dung. The soils were amended after two weeks of crude oil contamination. Soil samples were collected from the plastic bags for microbiological analyses. The isolates were cultured to test their ability to grow on crude oil. The ability of the isolates to utilize the crude oil was determined using media dilution technique. Bacteria such as Proteus mirabilis, Bacillus lacterosporus, Morganella morganii, Serratia marcescens and Bacillus alvei were isolated. The variables measured were heterotrophic bacterial populations, hydrocarbon utilizing bacterial populations and the percentage of crude oil degraded in the soils. Data collected were subjected to analysis of variance (ANOVA). Results obtained indicated that all the different weights of cow dung showed appreciable effect in crude oil decontamination. Based on the findings of the experiments, it could be deduced that 120g of cow dung promoted higher degradation of hydrocarbons. Thus, it should be recommended for remediation of crude oil contaminated soil in the study area.

Keywords: crude oil, cow dung, amendment, bioremediation, decontamination

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Authors: Wei Tian, Jie Liang, Hammad Naveed

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Beta-barrel membrane proteins are found in the outer membrane of gram-negative bacteria, mitochondria, and chloroplasts. They carry out diverse biological functions, including pore formation, membrane anchoring, enzyme activity, and bacterial virulence. In addition, beta-barrel membrane proteins increasingly serve as scaffolds for bacterial surface display and nanopore-based DNA sequencing. Due to difficulties in experimental structure determination, they are sparsely represented in the protein structure databank and computational methods can help to understand their biophysical principles. We have developed a novel computational method to predict the 3D structure of beta-barrel membrane proteins using evolutionary coupling (EC) constraints and a reduced state space. Combined with an empirical potential function, we can successfully predict strand register at > 80% accuracy for a set of 49 non-homologous proteins with known structures. This is a significant improvement from previous results using EC alone (44%) and using empirical potential function alone (73%). Our method is general and can be applied to genome-wide structural prediction.

Keywords: beta-barrel membrane proteins, structure prediction, evolutionary constraints, reduced state space

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Authors: Ilknur Tuncer, Nihayet Bizsel

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The scarcity of research in bacterial diversity and antimicrobial resistance in coastal environments as in Turkish coasts leads to difficulties in developing efficient monitoring and management programs. In the present study, biogeochemical analysis of sediments and antimicrobial susceptibility analysis of bacteria in Izmir Bay, eastern Aegean Sea under high anthropogenic pressure were aimed in summer period when anthropogenic input was maximum and at intertidal zone where the first terrigenious contact occurred for aquatic environment. Geochemical content of the intertidal zone of Izmir Bay was firstly illustrated such that total and organic carbon, nitrogen and phosphorus contents were high and the grain size distribution varied as sand and gravel. Bacterial diversity and antibiotic resistance were also firstly given for Izmir Bay. Antimicrobially assayed isolates underlined the multiple resistance in the inner, middle and outer bays with overall 19% high MAR (multiple antibiotic resistance) index. Phylogenetic analysis of 16S rRNA gene sequences indicated that 67 % of isolates belonged to the genus Bacillus and the rest included the families Alteromonadaceae, Bacillaceae, Exiguobacteriaceae, Halomonadaceae, Planococcaceae, and Staphylococcaceae.

Keywords: bacterial phylogeny, multiple antibiotic resistance, 16S rRNA genes, Izmir Bay, Aegean Sea

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Authors: Rupa Rani, Vipin Kumar

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Endosulfan, an organochlorine pesticide is of environmental concern due to its apparent persistence and toxicity. It has been reported as contaminants in soil, air, and water and is bioaccumulated and magnified in ecosystems. The combined use of microorganisms and plants has great potential for remediating soil contaminated with organic compounds such as pesticides. The objective of this study was to evaluate whether the bacterial inoculation influences plant growth promotion, endosulfan degradation in soil and endosulfan accumulation in different plant parts. Lycopersicon esculentum L. (Tomato) was grown in endosulfan spiked soil and inoculated with endosulfan tolerant bacterial strains. Endosulfan residues from different parts of plants and soil were extracted and estimated by using gas chromatograph equipped with 63Ni electron capture detector (GC-ECD). The inoculation of bacterial strains into the soil with plants showed a beneficial effect on endosulfan degradation and plant biomass production. Maximum endosulfan (90%) degradation was observed after 120 days of bacterial inoculation in the soil. Furthermore, there was significantly less endosulfan accumulation in roots and shoots of bacterial strains inoculated plants as compared to uninoculated plants. The results show the effectiveness of inoculated endosulfan tolerant bacterial strains to increase the remediation of endosulfan contaminated soil.

Keywords: organochlorine pesticides, endosulfan, degradation, plant-bacteria partnerships

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Authors: Laura Norfolk, Georgina Zimbitas, Jan Sefcik, Sarah Staniland

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Magnetite nanoparticles (MNP) have been an area of increasing research interest due to their extensive applications in industry, such as in carbon capture, water purification, and crucially, the biomedical industry. The use of MNP in the biomedical industry is rising, with studies on their effect as Magnetic resonance imaging contrast agents, drug delivery systems, and as hyperthermic cancer treatments becoming prevalent in the nanomaterial research community. Particles used for biomedical purposes must meet stringent criteria; the particles must have consistent shape and size between particles. Variation between particle morphology can drastically alter the effective surface area of the material, making it difficult to correctly dose particles that are not homogeneous. Particles of defined shape such as octahedral and cubic have been shown to outperform irregular shaped particles in some applications, leading to the need to synthesize particles of defined shape. In nature, highly homogeneous MNP are found within magnetotactic bacteria, a unique bacteria capable of producing magnetite nanoparticles internally under ambient conditions. Biomineralisation proteins control the properties of the MNPs, enhancing their homogeneity. One of these proteins, Mms6, has been successfully isolated and used in vitro as an additive in room-temperature co-precipitation reactions (RTCP) to produce particles of defined mono-dispersed size & morphology. When considering future industrial scale-up it is crucial to consider the costs and feasibility of an additive, as an additive that is not readily available or easily synthesized at a competitive price will not be sustainable. As such, additives selected for this research are inspired by the functional groups of biomineralisation proteins, but cost-effective, environmentally friendly, and compatible with scale-up. Diethylenetriamine (DETA), triethylenetetramine (TETA), tetraethylenepentamine (TEPA), and pentaethylenehexamine (PEHA) have been successfully used in RTCP to modulate the properties of particles synthesized, leading to the formation of octahedral nanoparticles with no use of organic solvents, heating, or toxic precursors. By extending this principle to a fluidic system, ongoing research will reveal whether the amine additives can also exert morphological control in an environment which is suited toward higher particle yield. Two fluidic systems have been employed; a peristaltic turbulent flow mixing system suitable for the rapid production of MNP, and a macrofluidic system for the synthesis of tailored nanomaterials under a laminar flow regime. The presence of the amine additives in the turbulent flow system in initial results appears to offer similar morphological control as observed under RTCP conditions, with higher proportions of octahedral particles formed. This is a proof of concept which may pave the way to green synthesis of tailored MNP on an industrial scale. Mms6 and amine additives have been used in the macrofluidic system, with Mms6 allowing magnetite to be synthesized at unfavourable ferric ratios, but no longer influencing particle size. This suggests this synthetic technique while still benefiting from the addition of additives, may not allow additives to fully influence the particles formed due to the faster timescale of reaction. The amine additives have been tested at various concentrations, the results of which will be discussed in this paper.

Keywords: bioinspired, green synthesis, fluidic, magnetite, morphological control, scale-up

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Authors: M. J. Norashirene, Y. Zakiah, S. Nurdiana, I. Nur Hilwani, M. H. Siti Khairiyah, M. J. Muhamad Arif

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In this study, six bacterial isolates of a slightly thermophilic organism from the Sg. Klah hot spring, Malaysia were successfully isolated and designated as M7T55D1, M7T55D2, M7T55D3, M7T53D1, M7T53D2 and M7T53D3 respectively. The bacterial isolates were screened for their cellulose hydrolytic ability on Carboxymethlycellulose agar medium. The isolated bacterial strains were identified morphologically, biochemically and molecularly with the aid of 16S rDNA sequencing. All of the bacteria showed their optimum growth at a slightly alkaline pH of 7.5 with a temperature of 55°C. All strains were Gram-negative, non-spore forming type, strictly aerobic, catalase-positive and oxidase-positive with the ability to produce thermostable cellulase. Based on BLASTn results, bacterial isolates of M7T55D2 and M7T53D1 gave the highest homology (97%) with similarity to Tepidimonas ignava while isolates M7T55D1, M7T55D3, M7T53D2 and M7T53D3 showed their closest homology (97%-98%) with Tepidimonas thermarum. These cellulolytic thermophiles might have a commercial potential to produce valuable thermostable cellulase.

Keywords: cellulase, cellulolytic, thermophiles, 16S rDNA gene

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Authors: Danyi Wang, Andrew Schriefer, Dennis O'Rourke, Brajendra Kumar, Yang Liu, Fei Zhong, Juergen Scheuenpflug, Zheng Feng

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Purpose: It has been recognized that the microbiome plays critical roles in disease pathogenesis, including cancer, autoimmune disease, and multiple sclerosis. To develop a clinical-grade assay for exploring microbiome-derived clinical biomarkers across disease areas, a two-phase approach is implemented. 1) Identification of the optimal sample preparation reagents using pre-mixed bacteria and healthy donor stool samples coupled with proprietary Sigma-Aldrich® bioinformatics solution. 2) Exploratory analysis of patient samples for enabling precision medicine. Study Procedure: In phase 1 study, we first compared the 16S sequencing results of two ATCC® microbiome standards (MSA 2002 and MSA 2003) across five different extraction kits (Kit A, B, C, D & E). Both microbiome standards samples were extracted in triplicate across all extraction kits. Following isolation, DNA quantity was determined by Qubit assay. DNA quality was assessed to determine purity and to confirm extracted DNA is of high molecular weight. Bacterial 16S ribosomal ribonucleic acid (rRNA) amplicons were generated via amplification of the V3/V4 hypervariable region of the 16S rRNA. Sequencing was performed using a 2x300 bp paired-end configuration on the Illumina MiSeq. Fastq files were analyzed using the Sigma-Aldrich® Microbiome Platform. The Microbiome Platform is a cloud-based service that offers best-in-class 16S-seq and WGS analysis pipelines and databases. The Platform and its methods have been extensively benchmarked using microbiome standards generated internally by MilliporeSigma and other external providers. Data Summary: The DNA yield using the extraction kit D and E is below the limit of detection (100 pg/µl) of Qubit assay as both extraction kits are intended for samples with low bacterial counts. The pre-mixed bacterial pellets at high concentrations with an input of 2 x106 cells for MSA-2002 and 1 x106 cells from MSA-2003 were not compatible with the kits. Among the remaining 3 extraction kits, kit A produced the greatest yield whereas kit B provided the least yield (Kit-A/MSA-2002: 174.25 ± 34.98; Kit-A/MSA-2003: 179.89 ± 30.18; Kit-B/MSA-2002: 27.86 ± 9.35; Kit-B/MSA-2003: 23.14 ± 6.39; Kit-C/MSA-2002: 55.19 ± 10.18; Kit-C/MSA-2003: 35.80 ± 11.41 (Mean ± SD)). Also, kit A produced the greatest yield, whereas kit B provided the least yield. The PCoA 3D visualization of the Weighted Unifrac beta diversity shows that kits A and C cluster closely together while kit B appears as an outlier. The kit A sequencing samples cluster more closely together than both the other kits. The taxonomic profiles of kit B have lower recall when compared to the known mixture profiles indicating that kit B was inefficient at detecting some of the bacteria. Conclusion: Our data demonstrated that the DNA extraction method impacts DNA concentration, purity, and microbial communities detected by next-generation sequencing analysis. Further microbiome analysis performance comparison of using healthy stool samples is underway; also, colorectal cancer patients' samples will be acquired for further explore the clinical utilities. Collectively, our comprehensive qualification approach, including the evaluation of optimal DNA extraction conditions, the inclusion of positive controls, and the implementation of a robust qualified bioinformatics pipeline, assures accurate characterization of the microbiota in a complex matrix for deciphering the deep biology and enabling precision medicine.

Keywords: 16S rRNA sequencing, analytical validation, bioinformatics pipeline, metagenomics

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