Search results for: bacterial and fungal strains

Authors: Cora Fernandez Dacosta, John A. Posada, Andrea Ramirez

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The biodegradable family of polymers polyhydroxyalkanoates is an interesting substitute for convectional fossil-based plastics. However, the manufacturing and environmental impacts associated with their production via intracellular bacterial fermentation are strongly dependent on the raw material used and on energy consumption during the extraction process, limiting their potential for commercialization. Industrial wastewater is studied in this paper as a promising alternative feedstock for waste valorization. Based on results from laboratory and pilot-scale experiments, a conceptual process design, techno-economic analysis and life cycle assessment are developed for the large-scale production of the most common type of polyhydroxyalkanoate, polyhydroxbutyrate. Intracellular polyhydroxybutyrate is obtained via fermentation of microbial community present in industrial wastewater and the downstream processing is based on chemical digestion with surfactant and hypochlorite. The economic potential and environmental performance results help identifying bottlenecks and best opportunities to scale-up the process prior to industrial implementation. The outcome of this research indicates that the fermentation of wastewater towards PHB presents advantages compared to traditional PHAs production from sugars because the null environmental burdens and financial costs of the raw material in the bioplastic production process. Nevertheless, process optimization is still required to compete with the petrochemicals counterparts.

Keywords: Circular economy, life cycle assessment, polyhydroxyalkanoates, waste valorization.

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Authors: Jaruwan Chutrtong

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Yoghurt is a fermented milk product. The process of making yogurt involves fermenting milk with live and active bacterial cultures by adding bacteria directly to the dairy product. It is usually made with a culture of Lactobacillus sp. (L. acidophilus or L. bulgaricus) and Streptococcus thermophilus. Many people like to eat it plain or flavored and it's also use as ingredient in many dishes. Yogurt is rich in nutrients including the microorganism which have important role in balancing the digestion and absorption of the boy.Consumers will benefit from lactic acid bacteria more or less depending on the amount of bacteria that lives in yogurt while eating. When purchasing yogurt, consumers should always check the label for live cultures. Yoghurt must keep in refrigerator at 4°C for up to ten days. After this amount of time, the cultures often become weak. This research studied freezing dry yogurt storage by monitoring on the survival of microorganisms when stored at different temperatures. At 30⁰C, representative room temperature of country in equator zone, number of lactic acid bacteria reduced 4 log cycles in 10 week. At 40⁰C, representative temperature in summer of country in equator zone, number of lactic acid bacteria also dropped 4 log cycle in 10 week, similar as storage at 30⁰C. But drying yogurt storage at 40⁰C couldn’t reformed to be good character yogurt as good as storage at 40⁰C only 4 week storage too. After 1 month, it couldn’t bring back the yogurt form. So if it is inevitable to keep yogurt powder at a temperature of 40°C, yoghurt is maintained only up to 4 weeks.

Keywords: Dynamic, dry yoghurt, storage, temperature.

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

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

Keywords: Arbuscular mycorrhizal fungi, biocontrol methods, Phelipanche ramosa, F. oxisporum spp.

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Authors: Samy A. Selim

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The antimicrobial, antiplasmid and cytotoxic activities of marine algae Halimeda opuntia and Sarconema filiforme were investigated. Antimicrobial bioassay against some human pathogenic bacteria and yeast were conducted using disc diffusion method. Halimeda extract exhibited antibacterial activity against six species of microrganisms, with significant inhibition against Staphylococcus aureus. While Sarconema extract was better potent as antifungal against Candida albicans. Comparative antibacterial studies showed that Halimeda extract showed equivalent or better activity as compared with commercial antibiotic when tested against Staphylococcus aureus. Further tests conducted using dilution method showed both extracts as having bacteriostatic mode of action against the tested microorganisms. Methanol extract of two species showed significant cytotoxicity (LC50 <500μg) on brine shrimp. Halimeda opuntia showed highest cytotoxic activity (LC50 =192.3μg). Also, the present investigation was undertaken to investigate the ability of methanolic extract of the algal extracts to cure R-plasmids from certain clinical E. coli isolates. The active fraction of Halimeda and Sarconema could cure plasmids from E. coli at curing efficiencies of approximately 78%. The active fraction mediated plasmid curing resulted in the subsequent loss of antibiotic resistance encoded in the plasmids as revealed by antibiotic resistance profile of cured strains. The screening results confirm the possible use of marine algae Halimeda opuntia and Sarconema filiforme as a source of pharmacological benefits.

Keywords: Antimicrobial, antiplasmid Cytotoxicity, Marine Algae.

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Authors: Abobaker Mohammed Qasem Farhan, Shangming Yang, Mohammed Al-Nehari

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Pneumonia is a kind of lung disease that creates congestion in the chest. Such pneumonic conditions lead to loss of life due to the severity of high congestion. Pneumonic lung disease is caused by viral pneumonia, bacterial pneumonia, or COVID-19 induced pneumonia. The early prediction and classification of such lung diseases help reduce the mortality rate. We propose the automatic Computer-Aided Diagnosis (CAD) system in this paper using the deep learning approach. The proposed CAD system takes input from raw computerized tomography (CT) scans of the patient's chest and automatically predicts disease classification. We designed the Hybrid Deep Learning Algorithm (HDLA) to improve accuracy and reduce processing requirements. The raw CT scans are pre-processed first to enhance their quality for further analysis. We then applied a hybrid model that consists of automatic feature extraction and classification. We propose the robust 2D Convolutional Neural Network (CNN) model to extract the automatic features from the pre-processed CT image. This CNN model assures feature learning with extremely effective 1D feature extraction for each input CT image. The outcome of the 2D CNN model is then normalized using the Min-Max technique. The second step of the proposed hybrid model is related to training and classification using different classifiers. The simulation outcomes using the publicly available dataset prove the robustness and efficiency of the proposed model compared to state-of-art algorithms.

Keywords: CT scans, COVID-19, deep learning, image processing, pneumonia, lung disease.

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Authors: K. F. Khan, Samsuddin Khan

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The Paleoproterozoic phosphorite deposit of Sallopat block of Banswara district of Rajasthan belongs to kalinjara formation of lunavada group of Aravalli Super Group. The phosphorites are found to occur as massive, brecciated, laminated and stromatolitic associated with calcareous quartzite, interbedded dolomite and multi coloured chert. The phosphorites are showing alternate brown and grey coloured concentric rims which are composed of phosphate, calcite and quartz minerals. Petro-mineralogical studies of phosphorite samples using petrological microscope, XRD, FEG- SEM and EDX reveal that apatite-(CaF) and apatite-(CaOH) are phosphate minerals which are intermixed with minor amount of carbonate materials. Sporadic findings of the uniform tiny granules of partially anisotropic apatite-(CaF) along with dolomite, calcite, quartz, muscovite, zeolite and other gangue minerals have been observed with the replacement of phosphate material by quartz and carbonate. The presence of microbial filaments of organic matter and alternate concentric rims of stromatolitic structure may suggest that the deposition of the phosphate took place in shallow marine oxidizing environmental conditions leading to the formation of phosphorite layers as primary biogenic precipitates by bacterial or algal activities. Different forms and texture of phosphate minerals may be due to environmental vicissitudes at the time of deposition followed by some replacement processes and biogenic activities.

Keywords: Petro-mineralogy, phosphorites, sallopat, apatite.

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Authors: Ilma Robo, Saimir Heta, Greta Plaka, Vera Ostreni

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Purpose: Drug gingival hyperplasia is an uncommon pathology encountered during routine work in dental units. The purpose of this paper is to present the clinical appearance of gingival hyperplasia caused by medications. There are already three classes of medications that cause hyperplasia and based on data from the literature, the clinical cases encountered and included in this study have been compared. Materials and Methods: The study was conducted in a total of 311 patients, out of which 182 patients were included in our study, meeting the inclusion criteria. After each patient's history was recorded and it was found that patients were in their knowledge of chronic illness, undergoing treatment of gingivitis hypertrophic drugs was performed with a clinical examination of oral cavity and assessment by vertical and horizontal evaluation according to the periodontal indexes. Results: Of the data collected during the study, it was observed that 97% of patients with gingival hyperplasia are treated with nifedipine. 84% of patients treated with selected medicines and gingival hyperplasia in the oral cavity has been exposed at time period for more than 1 year and 1 month. According to the GOI, in the first rank of this index are about 21% of patients, in the second rank are 52%, in the third rank are 24% and in the fourth grade are 3%. According to the horizontal growth index of gingival hyperplasia, grade 1 included about 61% of patients and grade 2 included about 39% of patients with gingival hyperplasia. Bacterial index divides patients by degrees: grading 0 - 8.2%, grading 1 - 32.4%, grading 2 - 14% and grading 3 - 45.1%. Conclusions: The highest percentage of gingival hyperplasia caused by drugs is due to dosing of nifedipine for a duration of dosing and application for systemic healing for more than 1 year.

Keywords: Drug gingival hyperplasia, horizontal growth index, vertical growth index.

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Authors: Henry Munoz, Muhammad Mohsan, Takashi Kiyota

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Strength and deformability characteristics of a liquefiable sand deposit including the development of earthquake-induced shear stress and shear strain as well as soil softening via the progressive degradation of shear modulus were studied via shaking table experiments. To do so, a model of a liquefiable sand deposit was constructed and densely instrumented where accelerations, pressures, and displacements at different locations were continuously monitored. Furthermore, the confinement effects on the strength and deformation characteristics of the liquefiable sand deposit due to an external surcharge by placing a heavy concrete slab (i.e. the model of an actual structural rigid pavement) on the ground surface were examined. The results indicate that as the number of seismic-loading cycles increases, the sand deposit softens progressively as large shear strains take place in different sand elements. Liquefaction state is reached after the combined effects of the progressive degradation of the initial shear modulus associated with the continuous decrease in the mean principal stress, and the buildup of the excess of pore pressure takes place in the sand deposit. Finally, the confinement effects given by a concrete slab placed on the surface of the sand deposit resulted in a favorable increasing in the initial shear modulus, an increase in the mean principal stress and a decrease in the softening rate (i.e. the decreasing rate in shear modulus) of the sand, thus making the onset of liquefaction to take place at a later stage. This is, only after the sand deposit having a concrete slab experienced a higher number of seismic loading cycles liquefaction took place, in contrast to an ordinary sand deposit having no concrete slab.

Keywords: Liquefaction, shaking table, shear modulus degradation, earthquake.

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Authors: Astorga-Trejo Rebeca, Fonseca-Peralta Héctor Manuel, Beltrán-Arredondo Laura Ivonne, Castro-Martínez Claudia

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The use of biomass as feedstock for the production of fuels and other chemicals of interest is an ever growing accepted option in the way to the development of biorefinery complexes. In the Mexican state of Sinaloa, a significant amount of residues from corn crops are produced every year, most of which can be converted to bioethanol and other products through biotechnological conversion using yeast and other microorganisms. Therefore, the objective of this work was to take advantage of corn stover and evaluate its potential as a substrate for the production of second generation bioethanol (2G), enzymes and xylitol. To produce bioethanol 2G, an acid-alkaline pretreatment was carried out prior to saccharification and fermentation. The microorganisms used for the production of enzymes, as well as for the production of xylitol, were isolated and characterized in our work group. Statistical analysis was performed using Design Expert version 11.0. The results showed that it is possible to obtain 2G bioethanol employing corn stover as a carbon source and Saccharomyces cerevisiae ItVer01 and Candida intermedia CBE002 with yields of 0.42 g and 0.31 g, respectively. It was also shown that C. intermedia has the ability to produce xylitol with a good yield (0.46 g/g). On the other hand, qualitative and quantitative studies showed that the native strains of Fusarium equiseti (0.4 IU/mL - xylanase), Bacillus velezensis (1.2 IU/mL – xylanase and 0.4 UI/mL - amylase) and Penicillium funiculosum (1.5 IU/mL - cellulases) have the capacity to produce xylanases, amylases or cellulases using corn stover as raw material. This study allowed us to demonstrate that it is possible to use corn stover as a carbon source, a low-cost raw material with high availability in our country, to obtain bioproducts of industrial interest, using processes that are more environmentally friendly and sustainable. It is necessary to continue the optimization of each bioprocess.

Keywords: Biomass, corn stover, biorefinery, bioethanol 2G, enzymes, xylitol.

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Authors: Prasad Senadheera, Younousse Saidi, Frans JM Maathuis

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Rice seed expression (cDNA) library in the Lambda Zap 11® phage constructed from the developing grain 10-20 days after flowering was transformed into yeast for functional complementation assays in three salt sensitive yeast mutants S. cerevisiae strain CY162, G19 and Axt3K. Transformed cells of G19 and Axt3K with pYES vector with cDNA inserts showed enhance tolerance than those with empty pYes vector. Sequencing of the cDNA inserts revealed that they encode for the putative proteins with the sequence homologous to rice putative protein PROLM24 (Os06g31070), a prolamin precursor. Expression of this cDNA did not affect yeast growth in absence of salt. Axt3k and G19 strains expressing the PROLM24 were able to grow upto 400 mM and 600 mM of NaCl respectively. Similarly, Axt3k mutant with PROLM24 expression showed comparatively higher growth rate in the medium with excess LiCl (50 mM). The observation that expression of PROLM24 rescued the salt sensitive phenotypes of G19 and Axt3k indicates the existence of a regulatory system that ameliorates the effect of salt stress in the transformed yeast mutants. However, the exact function of the cDNA sequence, which shows partial sequence homology to yeast UTR1 is not clear. Although UTR1 involved in ferrous uptake and iron homeostasis in yeast cells, there is no evidence to prove its role in Na+ homeostasis in yeast cells. Absence of transmembrane regions in Os06g31070 protein indicates that salt tolerance is achieved not through the direct functional complementation of the mutant genes but through an alternative mechanism.

Keywords: Rice seed expression, salt stress, prolamin, salinitytolerance, Oryza sativa

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Authors: Jaruwan Chutrtong, Tanakwan Bussabun

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Study production of tempeh inoculums powder by freeze-drying comparison with dry at 50°C and the sun bask for developing efficient tempeh inoculums for tempeh producing. Rhizopus oligosporus in PDA slant cultures was incubated at 30^oC for 3-5 days until spores and mycelium. Preparation spores suspension with sterilized water and then count the number of started spores. Fill spores suspension in Rice flour and soy flour, mixed with water (in the ratio 10: 7), which is steamed and sterilized at 121°C 15min. Incubated at room temperature for 4 days, count number of spores. Then take the progressive infection and full spore dough to dry at 50°C, sun bask, and lyophilize. Grind to powder. Then pack in plastic bags, stored at 5°C. To investigate quality of inoculums which use different methods, tempeh was fermented every 4 weeks for 24 weeks of the experiment. The result found that rice flour is not suitable to use as raw material in the production of powdered spores.  Fungi can growth rarely. Less number of spores and requires more time than soy flour. For drying method, lyophilization is the least possible time. Samples from this method are very hard and very dark and harder to grind than other methods. Drying at 50°C takes longer time than lyophilization but can also set time use for drying. Character of the dry samples is hard solid and brown color, but can be grinded easier. The sun drying takes the longest time, can’t determine the exact time. When the spore powder was used to fermented tempeh immediately, product has similar characters as which use spores that was fresh prepared. The tempeh has normal quality. When spore powder stored at low temperature, tempeh from storage spore in weeks 4, 8 and 12 is still normal. Time spending in production was close to the production of fresh spores. After storage  spores for 16 and 20 weeks, tempeh is still normal but growth and sporulation were take longer time than usual (about 6 hours). At 24 week storage, fungal growth is not good, made tempeh looks inferior to normal color, also smell and texture.

Keywords: Freeze drying, preparation, spore powder, tempeh.

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Authors: Abu Salim Mustafa

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Rv3873 is a relatively large size protein (371 amino acids in length) and its gene is located in the immunodominant genomic region of difference (RD)1 that is present in the genome of Mycobacterium tuberculosis but deleted from the genomes of all the vaccine strains of Bacillus Calmette Guerin (BCG) and most other mycobacteria. However, when tested for cellular immune responses using peripheral blood mononuclear cells from tuberculosis patients and BCG-vaccinated healthy subjects, this protein was found to be a major stimulator of cell mediated immune responses in both groups of subjects. In order to further identify the sequence of immunodominant epitopes and explore their Human Leukocyte Antigen (HLA)-restriction for epitope recognition, 24 peptides (25-mers overlapping with the neighboring peptides by 10 residues) covering the sequence of Rv3873 were synthesized chemically using fluorenylmethyloxycarbonyl chemistry and tested in cell mediated immune responses. The results of these experiments helped in the identification of an immunodominant peptide P9 that was recognized by people expressing varying HLA-DR types. Furthermore, it was also predicted to be a promiscuous binder with multiple epitopes for binding to HLA-DR, HLA-DP and HLA-DQ alleles of HLA-class II molecules that present antigens to T helper cells, and to HLA-class I molecules that present antigens to T cytotoxic cells. In addition, the evaluation of peptide P9 using an immunogenicity predictor server yielded a high score (0.94), which indicated a greater probability of this peptide to elicit a protective cellular immune response. In conclusion, P9, a peptide with multiple epitopes and ability to bind several HLA class I and class II molecules for presentation to cells of the cellular immune response, may be useful as a peptide-based vaccine against tuberculosis.

Keywords: Mycobacterium tuberculosis, Rv3873, peptides, vaccine

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Authors: Munish Garg, Vinni Ahuja

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In the past few years, high consumption of soft drinks has attracted negative attention world-wide due to its possible adverse effects, leading the health conscious people to find alternative nutraceutical or herbal health drinks. In the present study, a nutraceutical soft drink was developed utilizing some easily available and well known traditional herbs having nutritional potential. The key ingredients were selected as bael, amla, lemon juice, ashwagandha and poppy seeds based on their household routine use in the summer with proven refreshing, cooling and energetic feeling since ages. After several trials made, the final composition of nutraceutical summer soft drink was selected as most suitable combination based on the taste, physicochemical, microbial and organoleptic point of view. The physicochemical analysis of the prepared drink found to contain optimum level of titratable acidity, total soluble solids and pH which were in accordance of the commercial recommendations. There were no bacterial colonies found in the product therefore found within limits. During the nine point’s hedonic scale sensory evaluation, the drink was strongly liked for colour, taste, flavour and texture. The formulation was found to contain flavonoids (80mg/100ml), phenolics (103mg/100ml), vitamin C (250mg/100ml) and has antioxidant potential (75.52%) apart from providing several other essential vitamins, minerals and healthy components. The developed nutraceutical drink provides an economical and feasible option for the consumers with very good taste combined with potential health benefits. The present drink is potentially capable to replace the synthetic soft drinks available in the market.

Keywords: Herbal drink, nutraceuticals, summer drink, antioxidant.

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Authors: Mingren Shi, Michael Renton

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There is a world-wide need for the development of sustainable management strategies to control pest infestation and the development of phosphine (PH3) resistance in lesser grain borer (Rhyzopertha dominica). Computer simulation models can provide a relatively fast, safe and inexpensive way to weigh the merits of various management options. However, the usefulness of simulation models relies on the accurate estimation of important model parameters, such as mortality. Concentration and time of exposure are both important in determining mortality in response to a toxic agent. Recent research indicated the existence of two resistance phenotypes in R. dominica in Australia, weak and strong, and revealed that the presence of resistance alleles at two loci confers strong resistance, thus motivating the construction of a two-locus model of resistance. Experimental data sets on purified pest strains, each corresponding to a single genotype of our two-locus model, were also available. Hence it became possible to explicitly include mortalities of the different genotypes in the model. In this paper we described how we used two generalized linear models (GLM), probit and logistic models, to fit the available experimental data sets. We used a direct algebraic approach generalized inverse matrix technique, rather than the traditional maximum likelihood estimation, to estimate the model parameters. The results show that both probit and logistic models fit the data sets well but the former is much better in terms of small least squares (numerical) errors. Meanwhile, the generalized inverse matrix technique achieved similar accuracy results to those from the maximum likelihood estimation, but is less time consuming and computationally demanding.

Keywords: mortality estimation, probit models, logistic model, generalized inverse matrix approach, pest control simulation

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Authors: O. Onoriode Emoyan, E. Eyitemi Akporhonor, Charles Otobrise

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Polycyclic Aromatic Hydrocarbons (PAHs) are formed mainly because of incomplete combustion of organic materials during industrial, domestic activities or natural occurrence. Their toxicity and contamination of terrestrial and aquatic ecosystem have been established. However, with limited validity index, previous research has focused on PAHs isomer pair ratios of variable physicochemical properties in source identification. The objective of this investigation was to determine the empirical validity of Pearson Correlation Coefficient (PCC) and Cluster Analysis (CA) in PAHs source identification along soil samples of different land uses. Therefore, 16 PAHs grouped, as Endocrine Disruption Substances (EDSs) were determined in 10 sample stations in top and sub soils seasonally. PAHs was determined the use of Varian 300 gas chromatograph interfaced with flame ionization detector. Instruments and reagents used are of standard and chromatographic grades respectively. PCC and CA results showed that the classification of PAHs along pyrolitic and petrogenic organics used in source signature is about the predominance PAHs in environmental matrix. Therefore, the distribution of PAHs in the studied stations revealed the presence of trace quantities of the vast majority of the sixteen PAHs, which may ultimately inhabit the actual source signature authentication. Therefore, factors to be considered when evaluating possible sources of PAHs could be; type and extent of bacterial metabolism, transformation products/substrates, and environmental factors such as salinity, pH, oxygen concentration, nutrients, light intensity, temperature, co-substrates, and environmental medium are hereby recommended as factors to be considered when evaluating possible sources of PAHs.

Keywords: Comparative correlation, kinetically, polynuclear aromatic hydrocarbons, thermodynamically- favored PAHs, sources evaluation.

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Authors: Nadia El Alami El Hassani, Soukaina Motia, Benachir Bouchikhi, Nezha El Bari

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Doxycycline (DXy) is a cycline antibiotic, most frequently prescribed to treat bacterial infections in veterinary medicine. However, its broad antimicrobial activity and low cost, lead to an intensive use, which can seriously affect human health. Therefore, its spread in the food products has to be monitored. The scope of this work was to synthetize a sensitive and very selective molecularly imprinted polymer (MIP) for DXy detection in honey samples. Firstly, the synthesis of this biosensor was performed by casting a layer of carboxylate polyvinyl chloride (PVC-COOH) on the working surface of a gold screen-printed electrode (Au-SPE) in order to bind covalently the analyte under mild conditions. Secondly, DXy as a template molecule was bounded to the activated carboxylic groups, and the formation of MIP was performed by a biocompatible polymer by the mean of polyacrylamide matrix. Then, DXy was detected by measurements of differential pulse voltammetry (DPV). A non-imprinted polymer (NIP) prepared in the same conditions and without the use of template molecule was also performed. We have noticed that the elaborated biosensor exhibits a high sensitivity and a linear behavior between the regenerated current and the logarithmic concentrations of DXy from 0.1 pg.mL⁻¹ to 1000 pg.mL⁻¹. This technic was successfully applied to determine DXy residues in honey samples with a limit of detection (LOD) of 0.1 pg.mL⁻¹ and an excellent selectivity when compared to the results of oxytetracycline (OXy) as analogous interfering compound. The proposed method is cheap, sensitive, selective, simple, and is applied successfully to detect DXy in honey with the recoveries of 87% and 95%. Considering these advantages, this system provides a further perspective for food quality control in industrial fields.

Keywords: Electrochemical sensor, molecular imprinted polymer, doxycycline, food control.

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Authors: S. D. El Wakil, J. Rice

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The aim of the current work was to employ the finite element method to model a slab, with a small hole across its width, undergoing plastic plane strain deformation. The computational model had, however, to be validated by comparing its results with those obtained experimentally. Since they were in good agreement, the finite element method can therefore be considered a reliable tool that can help gain better understanding of the mechanism of ductile failure in structural members having stress raisers. The finite element software used was ANSYS, and the PLANE183 element was utilized. It is a higher order 2-D, 8-node or 6-node element with quadratic displacement behavior. A bilinear stress-strain relationship was used to define the material properties, with constants similar to those of the material used in the experimental study. The model was run for several tensile loads in order to observe the progression of the plastic deformation region, and the stress concentration factor was determined in each case. The experimental study involved employing the visioplasticity technique, where a circular mesh (each circle was 0.5 mm in diameter, with 0.05 mm line thickness) was initially printed on the side of an aluminum slab having a small hole across its width. Tensile loading was then applied to produce a small increment of plastic deformation. Circles in the plastic region became ellipses, where the directions of the principal strains and stresses coincided with the major and minor axes of the ellipses. Next, we were able to determine the directions of the maximum and minimum shear stresses at the center of each ellipse, and the slip-line field was then constructed. We were then able to determine the stress at any point in the plastic deformation zone, and hence the stress concentration factor. The experimental results were found to be in good agreement with the analytical ones.

Keywords: Finite element method to model a slab, slab undergoing plastic deformation, stress distribution around a circular hole, visioplasticity.

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Authors: P. Murphy, D. Vasic, A. W. Gunaratne, T. Tugonon, M. Ison, C. Pagonis, E. T. Sitchon, A. Le Busque, T. J. Borody

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Irritable bowel syndrome (IBS) is a gastrointestinal disorder characterized by an alteration in bowel movements. There are three different types of IBS: diarrhea-predominant IBS (IBS-D), constipation-predominant IBS (IBS-C) and IBS with mixed bowel habit (IBS-M). Antimicrobials are increasingly being used as treatment for all types of IBS. Due to this increased use and subsequent success, the gut microbiome as a factor in the etiology of IBS is becoming more apparent. Accepted standard treatment has focused on IBS-C and involves either vancomycin or rifaximin. Here, we report on a cohort of 18 patients treated with both vancomycin and rifaximin for IBS-D. These patients’ records were reviewed retrospectively. In this cohort, patients were aged between 24-74 years (mean 44 years) and nine were female. At baseline all patients had diarrhea, four with mucus and one with blood. Other reported symptoms include abdominal pain (n = 11) bloating (n = 9), flatulence (n = 7), fatigue (n = 4) and nausea (n = 3). Patient’s treatments were personalized according to their symptom severity and tolerability and were treated with a combination of rifaximin (500-3000 mg/d) and vancomycin (500 mg-1500 mg/d) for an ongoing period. Follow-ups were conducted between 2-32 weeks. Of all patients, 89% reported improvement of at least 1 symptom, one reported no change and one patient’s symptoms got worse. The success of this combination treatment could be due to the different mechanisms of action undertaken by each medication. Vancomycin works by inhibiting the cell wall of the bacteria and rifaximin by inhibiting protein synthesis. This success in treatment validates the idea that IBS-D may be driven by a bacterial infection of the gastrointestinal microbiome. As IBS-D presents similarly to Clostridium difficile and symptom improvement can occur with the same treatment as Clostridium difficile of rifaximin and vancomycin, there is reason to suggest that the infectious agent could be an unidentified strain of Clostridium. Although these results offer some validity to the theory, more research is required.

Keywords: Clostridium difficile infection, diarrhea predominant irritable bowel syndrome, microbiome, vancomycin/rifaximin combination.

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Authors: Avita K. Marihal, K.S. Jagadeesh, Sarita Sinha

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Pentachlorophenol (PCP) is a polychlorinated aromatic compound that is widespread in industrial effluents and is considered to be a serious pollutant. Among the variety of industrial effluents encountered, effluents from tanning industry are very important and have a serious pollution potential. PCP is also formed unintentionally in effluents of paper and pulp industries. It is highly persistent in soils and is lethal to a wide variety of beneficial microorganisms and insects, human beings and animals. The natural processes that breakdown toxic chemicals in the environment have become the focus of much attention to develop safe and environmentfriendly deactivation technologies. Microbes and plants are among the most important biological agents that remove and degrade waste materials to enable their recycling in the environment. The present investigation was carried out with the aim of developing a microbial system for bioremediation of PCP polluted soils. A number of plant species were evaluated for their ability to tolerate different concentrations of pentachlorophenol (PCP) in the soil. The experiment was conducted for 30 days under pot culture conditions. The toxic effect of PCP on plants was studied by monitoring seed germination, plant growth and biomass. As the concentration of PCP was increased to 50 ppm, the inhibition of seed germination, plant growth and biomass was also increased. Although PCP had a negative effect on all plant species tested, maize and groundnut showed the maximum tolerance to PCP. Other tolerating crops included wheat, safflower, sunflower, and soybean. From the rhizosphere soil of the tolerant seedlings, as many as twenty seven PCP tolerant bacteria were isolated. From soybean, 8; sunflower, 3; safflower 8; maize 2; groundnut and wheat, 3 each isolates were made. They were screened for their PCP degradation potentials. HPLC analyses of PCP degradation revealed that the isolate MAZ-2 degraded PCP completely. The isolate MAZ-1 was the next best isolate with 90 per cent PCP degradation. These strains hold promise to be used in the bioremediation of PCP polluted soils.

Keywords: Biodegradation, pentachlorophenol, rhizobacteria.

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Authors: W. Assaad, D. Wilmink, H. R. Pasaribu, H. J. M. Geijselaers

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Shell has developed a mono-diameter well concept for oil and gas wells as opposed to the traditional telescopic well design. A Mono-diameter well design allows well to have a single inner diameter from the surface all the way down to reservoir to increase production capacity, reduce material cost and reduce environmental footprint. This is achieved by expansion of liners (casing string) concerned using an expansion tool (e.g. a cone). Since the well is drilled in stages and liners are inserted to support the borehole, overlap sections between consecutive liners exist which should be expanded. At overlap, the previously inserted casing which can be expanded or unexpanded is called the host casing and the newly inserted casing is called the expandable casing. When the cone enters the overlap section, an expandable casing is expanded against a host casing, a cured cement layer and formation. In overlap expansion, ironing or lengthening may appear instead of shortening in the expandable casing when the pressure exerted by the host casing, cured cement layer and formation exceeds a certain limit. This pressure is related to cement strength, thickness of cement layer, host casing material mechanical properties, host casing thickness, formation type and formation strength. Ironing can cause implications that hinder the deployment of the technology. Therefore, the understanding of ironing becomes essential. A physical model is built in-house to calculate expansion forces, stresses, strains and post expansion casing dimensions under different conditions. In this study, only free casing and overlap expansion of two casings are addressed while the cement and formation will be incorporated in future study. Since the axial strain can be predicted by the physical model, the onset of ironing can be confirmed. In addition, this model helps in understanding ironing and the parameters influencing it. Finally, the physical model is validated with Finite Element (FE) simulations and small-scale experiments. The results of the study confirm that high pressure leads to ironing when the casing is expanded in tension mode.

Keywords: Casing expansion, cement, formation, metal forming, plasticity, well design.

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Authors: Suman Chaudhary, Rinku Dhanker, Tanvi, Sneh Goyal

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Sulphur oxidizing bacteria (SOB) have marked their significant role in perspectives of maintaining healthy environment as researchers from all over the world tested and apply these in waste water treatment plants, bioleaching of heavy metals, deterioration of bridge structures, concrete and for bioremediation purposes, etc. Also, these SOB are well adapted in all kinds of environment ranging from normal soil, water habitats to extreme natural sources like geothermal areas, volcanic eruptions, black shale and acid rock drainage (ARD). SOB have been isolated from low pH environment of anthropogenic origin like acid mine drainage (AMD) and bioleaching heaps, hence these can work efficiently in different environmental conditions. Besides having many applications in field of environment science, they may be proven to be very beneficial in area of agriculture as sulphur is the fourth major macronutrients required for the growth of plants. More amount of sulphur is needed by pulses and oilseed crops with respect to the cereal grains. Due to continuous use of land for overproduction of more demanding sulphur utilizing crops and without application of sulphur fertilizers, its concentration is decreasing day by day, and thus, sulphur deficiency is becoming a great problem as it affects the crop productivity and quality. Sulphur is generally found in soils in many forms which are unavailable for plants (cannot be use by plants) like elemental sulphur, thiosulphate which can be taken up by bacteria and converted into simpler forms usable by plants by undergoing a series of transformations. So, keeping the importance of sulphur in view for various soil types, oilseed crops and role of microorganisms in making them available to plants, we made an effort to isolate, optimize, and characterize SOB. Three potential strains of bacteria were isolated, namely SSF7, SSA21, and SSS6, showing sulphate production of concentration, i.e. 2.268, 3.102, and 2.785 mM, respectively. Also, these were optimized for various culture conditions like carbon, nitrogen source, pH, temperature, and incubation time, and characterization was also done.

Keywords: Sulphur oxidizing bacteria, isolation, optimization, characterization, sulphate production.

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Authors: Rupinder Bakshi, Geeta Walia, Anita Gupta

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Urinary Tract Infections are considered as one of the most common bacterial infections with an estimated annual global incidence of 150 million. Antimicrobial drug resistance is one of the major threats due to wide spread usage of uncontrolled antibiotics. In this study, a total number of 9149 urine samples were collected from R.H Patiala and processed in the Department of Microbiology G. M. C Patiala (January 2013 to December 2013). Urine samples were inoculated on MacConkey’s and blood agar plates and incubated at 370C for 24 hrs. The organisms were identified by colony characters, Gram’s staining, and biochemical reactions. Antimicrobial susceptibility of the isolates was determined against various antimicrobial agents (Hi – Media Mumbai India) by Kirby Bauer DISK diffusion method on Muller Hinton agar plates. Maximum patients were in the age group of 21-30 yrs followed by 31-40 yrs. Males (34%) are less prone to urinary tract infections than females (66%). Culture was positive in 25% of the samples. Escherichia coli was the most common isolate 60.3% followed by Klebsiella pneumoniae 13.5%, Proteus spp. 9% and Staphylococcus aureus 7.6%. Most of the urinary isolates were sensitive to, carbepenems, Aztreonam, Amikacin, and Piperacillin + Tazobactum. All the isolates showed a good sensitivity towards Nitrofurantoin (82%). ESBL production was found to be 70.6% in Escherichia coli and 29.4% in Klebsiella pneumonia. Susceptibility of ESBL producers to Imipenem, Nitrofurantoin and Amikacin were found to be 100%, 76%, and 75% respectively. Uropathogens are increasingly showing resistance to many antibiotics making empiric management of outpatient UTIs challenging. Ampicillin, Cotrimoxazole and Ciprofloxacin should not be used in empiric treatment. Nitrofurantoin could be used in lower urinary tract infection. Knowledge of uropathogens and their antimicrobial susceptibility pattern in a geographical region will help in appropriate and judicious antibiotic usage in a health care setup.

Keywords: Urinary Tract Infection, UTI, antibiotic susceptibility pattern, ESBL.

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

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

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

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Authors: Samy A. Selim, Nashwa I. Hagag

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Abstract–The objectives of the current study are to determine the prevalence, etiological agents, drug susceptibility pattern and plasmid profile of Acinetobacter baumannii isolates from Hospital-Acquired Infections (HAI) at Community Hospital, Al Jouf Province, Saudi Arabia. A total of 1890 patients had developed infection during hospital admission and were included in the study. Among those who developed nosocomial infections, 15(9.4), 10(2.7) and 118 (12.7) had respiratory tract infection (RTI), blood stream infections (BSI) and urinary tract (UTI) respectively. A total of 268 bacterial isolates were isolated from nosocomial infection. S. aureus was reported in 23.5% for of the total isolates followed by Klebsiella pneumoniae (17.5%), E. coli (17.2%), P. aeruginosa (11.9%), coagulase negative staphylococcus (9%), A. baumannii (7.1%), Enterobacter spp. (3.4%), Citrobacter freundii (3%), Proteus mirabilis (2.6%), and Proteus vulgaris and Enterococcous faecalis (0.7%). Isolated organisms are multi-drug resistant, predominantly Gram-positive pathogens with a high incidence of methicillin-resistant S. aureus, extended spectrum beta lactamase and vancomycin resistant enterococci organisms. The RFLP (Fragment Length Polymorphisms) patterns of plasmid preparations from isolated A. baumannii isolates had altered RFLP patterns, possibly due to the presence of plasmid(s). Five A. baumannii isolates harbored plasmids all of which were not less than 2.71kbp in molecular weight. Hence, it showed that the gene coding for the isolates were located on the plasmid DNA while the remaining isolates which have no plasmid might showed gene coding for antibiotic resistance being located on chromosomal DNA. Nosocomial infections represent a current problem in Community Hospital, Al Jouf Province, Saudi Arabia. Problems associated with SSI include infection with multidrug resistant pathogens which are difficult to treat and are associated with increased mortality.

Keywords: Hospital-Acquired Infections, Acinetobacter baumannii, antibiotic resistance, plasmid profile, RFLP patterns, Al Jouf Province, Saudi Arabia

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Authors: S. V. Ukwungwu, A. J. Abbas, G. G. Nasr

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The increasing high price of natural gas and oil with attendant increase in energy demand on world markets in recent years has stimulated interest in recovering residual oil saturation across the globe. In order to meet the energy security, efforts have been made in developing new technologies of enhancing the recovery of oil and gas, utilizing techniques like CO₂ flooding, water injection, hydraulic fracturing, surfactant flooding etc. Surfactant flooding however optimizes production but poses risk to the environment due to their toxic nature. Amongst proven records that have utilized other type of bacterial in producing biosurfactants for enhancing oil recovery, this research uses a technique to combine biosurfactants that will achieve a scale of EOR through lowering interfacial tension/contact angle. In this study, three biosurfactants were produced from three Bacillus species from freeze dried cultures using sucrose 3 % (w/v) as their carbon source. Two of these produced biosurfactants were screened with the TEMCO Pendant Drop Image Analysis for reduction in IFT and contact angle. Interfacial tension was greatly reduced from 56.95 mN.m^-1 to 1.41 mN.m^-1 when biosurfactants in cell-free culture (Bacillus licheniformis) were used compared to 4. 83mN.m^-1 cell-free culture of Bacillus subtilis. As a result, cell-free culture of (Bacillus licheniformis) changes the wettability of the biosurfactant treatment for contact angle measurement to more water-wet as the angle decreased from 130.75^o to 65.17^o. The influence of microbial treatment on crushed rock samples was also observed by qualitative wettability experiments. Treated samples with biosurfactants remained in the aqueous phase, indicating a water-wet system. These results could prove that biosurfactants can effectively change the chemistry of the wetting conditions against diverse surfaces, providing a desirable condition for efficient oil transport in this way serving as a mechanism for EOR. The environmental friendly effect of biosurfactants applications for industrial purposes play important advantages over chemically synthesized surfactants, with various possible structures, low toxicity, eco-friendly and biodegradability.

Keywords: Bacillus, biosurfactant, enhanced oil recovery, residual oil, wettability.

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Authors: P. Slepicka, N. Slepickova Kasalkova, I. Michaljanicova, O. Nedela, Z. Kolska, V. Svorcik

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Polymers exposed to laser or plasma treatment or modified with different wet methods which enable the introduction of nanoparticles or biologically active species, such as amino-acids, may find many applications both as biocompatible or anti-bacterial materials or on the contrary, can be applied for a decrease in the number of cells on the treated surface which opens application in single cell units. For the experiments, two types of materials were chosen, a representative of non-biodegradable polymers, polyethersulphone (PES) and polyhydroxybutyrate (PHB) as biodegradable material. Exposure of solid substrate to laser well below the ablation threshold can lead to formation of various surface structures. The ripples have a period roughly comparable to the wavelength of the incident laser radiation, and their dimensions depend on many factors, such as chemical composition of the polymer substrate, laser wavelength and the angle of incidence. On the contrary, biopolymers may significantly change their surface roughness and thus influence cell compatibility. The focus was on the surface treatment of PES and PHB by pulse excimer KrF laser with wavelength of 248 nm. The changes of physicochemical properties, surface morphology, surface chemistry and ablation of exposed polymers were studied both for PES and PHB. Several analytical methods involving atomic force microscopy, gravimetry, scanning electron microscopy and others were used for the analysis of the treated surface. It was found that the combination of certain input parameters leads not only to the formation of optimal narrow pattern, but to the combination of a ripple and a wrinkle-like structure, which could be an optimal candidate for cell attachment. The interaction of different types of cells and their interactions with the laser exposed surface were studied. It was found that laser treatment contributes as a major factor for wettability/contact angle change. The combination of optimal laser energy and pulse number was used for the construction of a surface with an anti-cellular response. Due to the simple laser treatment, we were able to prepare a biopolymer surface with higher roughness and thus significantly influence the area of growth of different types of cells (U-2 OS cells).

Keywords: Polymer treatment, laser, periodic pattern, cell response.

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Authors: A. Heshmati, M. Y Alikhani, M. T. Godarzi, M. R. Sadeghimanesh

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Herbal essential oil and extracts are a good source of natural antioxidants and antimicrobial compounds. Hypericum is one of the potential sources of these compounds. In this study, the antioxidant and antimicrobial activity of essential oil and aqueous, methanol, ethanol, ethyl acetate and acetone extract of Hypericum scabrum was assessed. Flowers of Hypericum scabrum were collected from the surrounding mountains of Hamadan province and after drying in the shade, the essential oil of the plant was extracted by Clevenger and water, methanol, ethanol, ethyl acetate and acetone extract was obtained by maceration method. Essential oil compounds were identified using the GC-Mass. The Folin-Ciocalteau and aluminum chloride (AlCl₃) colorimetric method was used to measure the amount of phenolic acid and flavonoids, respectively. Antioxidant activity was evaluated using DPPH and FRAP. The minimum inhibitory concentration (MIC) and the minimum bacterial/fungicide concentration (MBC/MFC) of essential oil and extracts were evaluated against Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, Salmonella typhimurium, Aspergillus flavus and Candida albicans. The essential oil yield of was 0.35%, the lowest and highest extract yield was related to ethyl acetate and water extract. The most component of essential oil was α-Pinene (46.35%). The methanol extracts had the highest phenolic acid (95.65 ± 4.72 µg galic acid equivalent/g dry plant) and flavonoids (25.39 ± 2.73 µg quercetin equivalent/g dry plant). The percentage of DPPH radical inhibition showed positive correlation with concentrations of essential oil or extract. The methanol and ethanol extract had the highest DDPH radical inhibitory. Essential oil and extracts of Hypericum had antimicrobial activity against the microorganisms studied in this research. The MIC and MBC values for essential oils were in the range of 25-25.6 and 25-50 μg/mL, respectively. For the extracts, these values were 1.5625-100 and 3.125-100 μg/mL, respectively. Methanol extracts had the highest antimicrobial activity. Essential oil and extract of Hypericum scabrum, especially methanol extract, have proper antimicrobial and antioxidant activity, and it can be used to control the oxidation and inhibit the growth of pathogenic and spoilage microorganisms. In addition, it can be used as a substitute for synthetic antioxidant and antimicrobial compounds.

Keywords: Antimicrobial, antioxidant, extract, hypericum.

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Authors: Ibrahim M. S. Shnawa, Bashar A. H. E. Alsadi, Kalida K. Alniaem

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In the immunologic sense, clinical infection is a state of failure of the immune system to combat the pathogenic weapon of the bacteria invading the host. A motile gram negative vibroid organism associated with marked mono and poly nuclear cell responses was traced during the examination of a clinical material from an infected common carp Cyprinus carpio. On primary plate culture, growth was shown to be pure, dense population of an Aeromonas-like colony morphotype. The pure isolate was found to be; Aerobic, facultatively anaerobic, non-halophilic, grew at 0C, and 37C, oxidase positive utilizes glucose through fermentative pathway, resist 0/129 and novobiocin, produces alanine and lysine decarboxylases but non-producing ornithine dehydrolases. Tests for the in vitro determinants of pathogenicity has shown to be; Betahaemolytic onto blood agar, gelatinase, casienase and amylase producer. Three in vivo determinants of pathogenicity were tested as, the lethal dose fifty, the pathogenesis and pathogenicity. It was evident that 0.1 milliliter of the causal bacterial cell suspension of a density 1 x 107 CFU/ml injected intramuscularly into an average of 100gms fish toke five days incubation period, then at the day six morbidity and mortality were initiated. LD50 was recorded at the day 12 post-infection. Use of an LD50 doses to study the pathogenicity, reveals mononuclear and polynuclear cell responses, on examining the stained direct films of the clinical materials from the experimentally infected fish. Re-isolation tests confirm that the reisolant is same. The course of the infection in natural case was shown manifestation of; skin ulceration, haemorrhage and descaling. On evisceration, the internal organs were shown; congestion in the intestines, spleen and, air sacs. The induced infection showed a milder form of these manifestations. The grading of the virulence of this organism was virulent causing chronic course of infections as indicated from the pathogenesis and pathogenicity studies. Thus the infectious bacteria were consistent with Aeromonas hydrophila, and the infection was chronic.

Keywords: Piscan, inflammatory respnonse, pure culture, pathogen, chronic, infection.

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Authors: M. S. Alatas, H. D. Umucalilar

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SARA is a common and serious metabolic disorder in early lactation in dairy cattle and in finishing beef cattle, caused by diets with high inclusion of cereal grain. This experiment was performed to determine the efficacy of Megasphaera elsdenii, a major lactate-utilizing bacterium in prevention/treatment of SARA in vivo. In vivo experimentation, it was used eight ruminally cannulated rams and it was applied the rapid adaptation with the mixture of grain based on wheat (80% wheat, 20% barley) and barley (80% barley, 20% wheat). During the systematic adaptation, it was followed the probability of SARA formation by being measured the rumen pH with two hours intervals after and before feeding. After being evaluated the data, it was determined the ruminal pH ranged from 5.2-5.6 on the condition of feeding with 60 percentage of grain mixture based on barley and wheat, that assured the definite form of subacute acidosis. In four days SARA period, M. elsdenii (1010 cfu ml-1) was inoculated during the first two days. During the SARA period, it was observed the decrease of feed intake with M. elsdenii inoculation. Inoculation of M. elsdenii was caused to differentiation of rumen pH (P<0.0001), while it was found the pH level approximately 5.55 in animals applied the inoculation, it was 5.63 pH in other animals. It was observed that total VFA with the bacterium inoculation tended to change in terms of grain feed (P<0.07). It increased with the effect of total VFA inoculation in barley based diet, but it was more stabilized in wheat based diet. Bacterium inoculation increased the ratio of propionic acid (18.33%-21.38%) but it caused to decrease the butyric acid, and acetic/propionic acid. During the rapid adaptation, the concentration of lactic acid in the rumen liquid increased depending upon grain level (P<0.0001). On the other hand bacterium inoculation did not have an effect on concentration of lactic acid. M. elsdenii inoculation did not affect ruminal ammonia concentration. In the group that did not apply inoculation, the level of ruminal ammonia concentration was higher than the others applied inoculation. M. elsdenii inoculation did not changed protozoa count in barley-based diet whereas it decreased in wheat-based diet. When it is generally evaluated, it is seen that M. elsdenii inoculation has not a positive impact on rumen parameters. Therefore, to reveal the full impact of the inoculation with different strains, feedstuffs and animal groups, further research is required.

Keywords: In vivo, subactute ruminal acidosis, Megasphaera elsdenii, rumen fermentation.

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Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

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Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast.

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