Search results for: bacterial aggregation
865 Microbiological Assessment of Soft Cheese (Wara), Raw Milk and Dairy Drinking Water from Selected Farms in Ido, Ibadan, Nigeria
Authors: Blessing C. Nwachukwu, Michael O. Taiwo, Wasiu A. Abibu, Isaac O. Ayodeji
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Milk is an important source of micro and macronutrients for humans. Soft Cheese (Wara) is an example of a by-product of milk. In addition, water is considered as one of the most vital resources in cattle farms. Due to the high consumption rate of milk and soft cheese and the traditional techniques involved in their production in Nigeria, there was a need for a microbiological assessment which will be of utmost public health importance. The study thus investigated microbial risk assessments associated with consumption of milk and soft cheese (Wara). It also investigated common pathogens present in dairy water in farms and antibiotic sensitivity profiling for implicated pathogens were conducted. Samples were collected from three different Fulani dairy herds in Ido local government, Ibadan, Oyo State, Nigeria and subjected to microbiological evaluation and antimicrobial susceptibility testing. Aspergillus flavus was the only isolated fungal isolate from Wara while Staphylococcus aureus, Vibro cholera, Hafnia alvei, Proteus mirabilis, Escherishia coli, Psuedomonas aeuroginosa, Citrobacter freundii, and Klebsiella pneumonia were the bacteria genera isolated from Wara, dairy milk and dairy drinking water. Bacterial counts from Wara from the three selected farms A, B and C were 3.5×105 CFU/ml, 4.0×105 CFU/ml and 5.3×105 CFU/ml respectively while the fungal count was 3CFU/100µl. The total bacteria count from dairy milk from the three selected farms A, B and C were Farms 2.0 ×105 CFU/ml, 3.5 × 105 CFU/ml and 6.5 × 105 CFU/ml respectively. 1.4×105 CFU/ml, 1.9×105 CFU/ml and 4.9×105 CFU/ml were the recorded bacterial counts from dairy water from farms A, B and C respectively. The highest antimicrobial resistance of 100% was recorded in Wara with Enrofloxacin, Gentamycin, Cefatriaxone and Colistin. The highest antimicrobial susceptibility of 100% was recorded in Raw milk with Enrofloxacin and Gentamicin. Highest antimicrobial intermediate response of 100% was recorded in Raw milk with Streptomycin. The study revealed that most of the cheeses sold at Ido local Government are contaminated with pathogens. Further research is needed on standardizing the production method to prevent pathogens from gaining access. The presence of bacteria in raw milk indicated contamination due to poor handling and unhygienic practices. Thus, drinking unpasteurized milk is hazardous as it increases the risk of zoonoses. Also, the Provision of quality drinking water is crucial for optimum productivity of dairy. Health education programs aiming at increasing awareness of the importance of clean water for animal health will be helpful.Keywords: dairy, raw milk, soft cheese, Wara
Procedia PDF Downloads 180864 Synthesis of Star Compounds Bearing a Porphyrin Core and Cholic Acid Units by Using Click Chemistry: Study of the Optical Properties and Aggregation
Authors: Edgar Aguilar-Ortíz, Nicolas Lévaray, Mireille Vonlanthen, Eric G. Morales-Espinoza, Ernesto Rivera, Xiao Xia Zhu
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Four new star compounds bearing a porphyrin core and cholic acid units, (TPPh(Zn) tetra-CA, TPPh(2H) tetra-CA, TPPh(Zn) octa-CA and TPPh(2H) octa-CA), have been synthesized using the Click Chemistry approach, which consist on azide-alkyne couplings. These novel functionalized porphyrins were characterized by 1H and 13C NMR spectroscopy and their structure was confirmed by MALDI-TOF. The optical properties of these compounds were studied by absorption and fluorescence spectroscopy. On the other hand, order to evaluate the amphiphilic properties of the cholic acid units combined with the optical response of the porphyrin core, we performed absorption and fluorescence studies in function of the polarity of the environment. It was found that as soon as we increase the polarity of the solvent, the Zn-metallated porphyrins, (TPPh(Zn) tetra-CA and TPPh(Zn) octa-CA), are able to form J aggregates, whereas the free-base porphyrins, TPPh(2H) tetra-CA and TPPh(2H) octa-CA, behaved differently.Keywords: aggregates, amphiphilic, cholic acid, click-chemistry, porphyrin
Procedia PDF Downloads 302863 Maackiain Attenuates Alpha-Synuclein Accumulation and Improves 6-OHDA-Induced Dopaminergic Neuron Degeneration in Parkinson's Disease Animal Model
Authors: Shao-Hsuan Chien, Ju-Hui Fu
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Parkinson’s disease (PD) is a degenerative disorder of the central nervous system that is characterized by progressive loss of dopaminergic neurons in the substantia nigra pars compacta and motor impairment. Aggregation of α-synuclein in neuronal cells plays a key role in this disease. At present, therapeutics for PD provides moderate symptomatic benefit but is not able to delay the development of this disease. Current efforts for the treatment of PD are to identify new drugs that show slow or arrest progressive course of PD by interfering with a disease-specific pathogenetic process in PD patients. Maackiain is a bioactive compound isolated from the roots of the Chinese herb Sophora flavescens. The purpose of the present study was to assess the potential for maackiain to ameliorate PD in Caenorhabditis elegans models. Our data reveal that maackiain prevents α-synuclein accumulation in the transgenic Caenorhabditis elegans model and also improves dopaminergic neuron degeneration, food-sensing behavior, and life-span in 6-hydroxydopamine-induced Caenorhabditis elegans model, thus indicating its potential as a candidate antiparkinsonian drug.Keywords: maackiain, Parkinson’s disease, dopaminergic neurons, α-Synuclein
Procedia PDF Downloads 197862 Characterizing and Developing the Clinical Grade Microbiome Assay with a Robust Bioinformatics Pipeline for Supporting Precision Medicine Driven Clinical Development
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
Procedia PDF Downloads 168861 Immunostimulatory Response of Supplement Feed in Fish against Aeromonas hydrophila
Authors: Shikha Rani, Neeta Sehgal, Vipin Kumar Verma, Om Prakash
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Introduction: Fish is an important protein source for humans and has great economic value. Fish cultures are affected due to various anthropogenic activities that lead to bacterial and viral infections. Aeromonas hydrophila is a fish pathogenic bacterium that causes several aquaculture outbreaks throughout the world and leads to huge mortalities. In this study, plants of no commercial value were used to investigate their immunostimulatory, antioxidant, anti-inflammatory, anti-bacterial, and disease resistance potential in fish against Aeromonas hydrophila, through fish feed fortification. Methods: The plant was dried at room temperature in the shade, dissolved in methanol, and analysed for biological compounds through GC-MS/MS. DPPH, FRAP, Phenolic, and flavonoids were estimated following standardized protocols. In silico molecular docking was also performed to validate its broad-spectrum activities based on binding affinity with specific proteins. Fish were divided into four groups (n=6; total 30 in a group): Group 1, non-challenged fish (fed on a non-supplemented diet); Group 2, fish challenged with bacteria (fed on a non-supplemented diet); Group 3 and 4, fish challenged with bacteria (A. hydrophila) and fed on plant supplemented feed at 2.5% and 5%. Blood was collected from the fish on 0, 7th, 14th, 21st, and 28th days. Serum was separated for glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase assay (ALP), lysozyme activity assay, superoxide dismutase assay (SOD), lipid peroxidation assay (LPO) and molecular parameters (including cytokine levels) were estimated through ELISA. The phagocytic activity of macrophages from the spleen and head kidney, along with quantitative analysis of immune-related genes, were analysed in different tissue samples. The digestive enzymes (Pepsin, Trypsin, and Chymotrypsin) were also measured to evaluate the effect of plant-supplemented feed on freshwater fish. Results and Discussion: GC-MS/MS analysis of a methanolic extract of plant validated the presence of key compounds having antioxidant, anti-inflammatory, anti-bacterial, anti-inflammatory, and immunomodulatory activities along with disease resistance properties. From biochemical investigations like ABTS, DPPH, and FRAP, the amount of total flavonoids, phenols, and promising binding affinities towards different proteins in molecular docking analysis helped us to realize the potential of this plant that can be used for investigation in the supplemented feed of fish. Measurement liver function tests, ALPs, oxidation-antioxidant enzyme concentrations, and immunoglobulin concentrations in the experimental groups (3 and 4) showed significant improvement as compared to the positive control group. The histopathological evaluation of the liver, spleen, and head kidney supports the biochemical findings. The isolated macrophages from the group fed on supplemented feed showed a higher percentage of phagocytosis and a phagocytic index, indicating an enhanced cell-mediated immune response. Significant improvements in digestive enzymes were also observed in fish fed on supplemented feed, even after weekly challenges with bacteria. Hence, the plant-fortified feed can be recommended as a regular feed to enhance fish immunity and disease resistance against the Aeromonas hydrophila infection after confirmation from the field trial.Keywords: immunostimulation, antipathogen, plant fortified feed, macrophages, GC-MS/MS, in silico molecular docking
Procedia PDF Downloads 82860 A Mixed Expert Evaluation System and Dynamic Interval-Valued Hesitant Fuzzy Selection Approach
Authors: Hossein Gitinavard, Mohammad Hossein Fazel Zarandi
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In the last decades, concerns about the environmental issues lead to professional and academic efforts on green supplier selection problems. In this sake, one of the main issues in evaluating the green supplier selection problems, which could increase the uncertainty, is the preferences of the experts' judgments about the candidate green suppliers. Therefore, preparing an expert system to evaluate the problem based on the historical data and the experts' knowledge can be sensible. This study provides an expert evaluation system to assess the candidate green suppliers under selected criteria in a multi-period approach. In addition, a ranking approach under interval-valued hesitant fuzzy set (IVHFS) environment is proposed to select the most appropriate green supplier in planning horizon. In the proposed ranking approach, the IVHFS and the last aggregation approach are considered to margin the errors and to prevent data loss, respectively. Hence, a comparative analysis is provided based on an illustrative example to show the feasibility of the proposed approach.Keywords: green supplier selection, expert system, ranking approach, interval-valued hesitant fuzzy setting
Procedia PDF Downloads 326859 Oral Microbiota as a Novel Predictive Biomarker of Response To Immune Checkpoint Inhibitors in Advanced Non-small Cell Lung Cancer Patients
Authors: Francesco Pantano, Marta Fogolari, Michele Iuliani, Sonia Simonetti, Silvia Cavaliere, Marco Russano, Fabrizio Citarella, Bruno Vincenzi, Silvia Angeletti, Giuseppe Tonini
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Background: Although immune checkpoint inhibitors (ICIs) have changed the treatment paradigm of non–small cell lung cancer (NSCLC), these drugs fail to elicit durable responses in the majority of NSCLC patients. The gut microbiota, able to regulate immune responsiveness, is emerging as a promising, modifiable target to improve ICIs response rates. Since the oral microbiome has been demonstrated to be the primary source of bacterial microbiota in the lungs, we investigated its composition as a potential predictive biomarker to identify and select patients who could benefit from immunotherapy. Methods: Thirty-five patients with stage IV squamous and non-squamous cell NSCLC eligible for an anti-PD-1/PD-L1 as monotherapy were enrolled. Saliva samples were collected from patients prior to the start of treatment, bacterial DNA was extracted using the QIAamp® DNA Microbiome Kit (QIAGEN) and the 16S rRNA gene was sequenced on a MiSeq sequencing instrument (Illumina). Results: NSCLC patients were dichotomized as “Responders” (partial or complete response) and “Non-Responders” (progressive disease), after 12 weeks of treatment, based on RECIST criteria. A prevalence of the phylum Candidatus Saccharibacteria was found in the 10 responders compared to non-responders (abundance 5% vs 1% respectively; p-value = 1.46 x 10-7; False Discovery Rate (FDR) = 1.02 x 10-6). Moreover, a higher prevalence of Saccharibacteria Genera Incertae Sedis genus (belonging to the Candidatus Saccharibacteria phylum) was observed in "responders" (p-value = 6.01 x 10-7 and FDR = 2.46 x 10-5). Finally, the patients who benefit from immunotherapy showed a significant abundance of TM7 Phylum Sp Oral Clone FR058 strain, member of Saccharibacteria Genera Incertae Sedis genus (p-value = 6.13 x 10-7 and FDR=7.66 x 10-5). Conclusions: These preliminary results showed a significant association between oral microbiota and ICIs response in NSCLC patients. In particular, the higher prevalence of Candidatus Saccharibacteria phylum and TM7 Phylum Sp Oral Clone FR058 strain in responders suggests their potential immunomodulatory role. The study is still ongoing and updated data will be presented at the congress.Keywords: oral microbiota, immune checkpoint inhibitors, non-small cell lung cancer, predictive biomarker
Procedia PDF Downloads 95858 Biodegradation of Chlorophenol Derivatives Using Macroporous Material
Authors: Dmitriy Berillo, Areej K. A. Al-Jwaid, Jonathan L. Caplin, Andrew Cundy, Irina Savina
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Chlorophenols (CPs) are used as a precursor in the production of higher CPs and dyestuffs, and as a preservative. Contamination by CPs of the ground water is located in the range from 0.15-100mg/L. The EU has set maximum concentration limits for pesticides and their degradation products of 0.1μg/L and 0.5μg/L, respectively. People working in industries which produce textiles, leather products, domestic preservatives, and petrochemicals are most heavily exposed to CPs. The International Agency for Research on Cancers categorized CPs as potential human carcinogens. Existing multistep water purification processes for CPs such as hydrogenation, ion exchange, liquid-liquid extraction, adsorption by activated carbon, forward and inverse osmosis, electrolysis, sonochemistry, UV irradiation, and chemical oxidation are not always cost effective and can cause the formation of even more toxic or mutagenic derivatives. Bioremediation of CPs derivatives utilizing microorganisms results in 60 to 100% decontamination efficiency and the process is more environmentally-friendly compared with existing physico-chemical methods. Microorganisms immobilized onto a substrate show many advantages over free bacteria systems, such as higher biomass density, higher metabolic activity, and resistance to toxic chemicals. They also enable continuous operation, avoiding the requirement for biomass-liquid separation. The immobilized bacteria can be reused several times, which opens the opportunity for developing cost-effective processes for wastewater treatment. In this study, we develop a bioremediation system for CPs based on macroporous materials, which can be efficiently used for wastewater treatment. Conditions for the preparation of the macroporous material from specific bacterial strains (Pseudomonas mendocina and Rhodococus koreensis) were optimized. The concentration of bacterial cells was kept constant; the difference was only the type of cross-linking agents used e.g. glutaraldehyde, novel polymers, which were utilized at concentrations of 0.5 to 1.5%. SEM images and rheology analysis of the material indicated a monolithic macroporous structure. Phenol was chosen as a model system to optimize the function of the cryogel material and to estimate its enzymatic activity, since it is relatively less toxic and harmful compared to CPs. Several types of macroporous systems comprising live bacteria were prepared. The viability of the cross-linked bacteria was checked using Live/Dead BacLight kit and Laser Scanning Confocal Microscopy, which revealed the presence of viable bacteria with the novel cross-linkers, whereas the control material cross-linked with glutaraldehyde(GA), contained mostly dead cells. The bioreactors based on bacteria were used for phenol degradation in batch mode at an initial concentration of 50mg/L, pH 7.5 and a temperature of 30°C. Bacterial strains cross-linked with GA showed insignificant ability to degrade phenol and for one week only, but a combination of cross-linking agents illustrated higher stability, viability and the possibility to be reused for at least five weeks. Furthermore, conditions for CPs degradation will be optimized, and the chlorophenol degradation rates will be compared to those for phenol. This is a cutting-edge bioremediation approach, which allows the purification of waste water from sustainable compounds without a separation step to remove free planktonic bacteria. Acknowledgments: Dr. Berillo D. A. is very grateful to Individual Fellowship Marie Curie Program for funding of the research.Keywords: bioremediation, cross-linking agents, cross-linked microbial cell, chlorophenol degradation
Procedia PDF Downloads 212857 Green-synthesized of Selenium Nanoparticles Using Garlic Extract and Their Application for Rapid Detection of Salicylic Acid in Milk
Authors: Kashif Jabbar
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Milk adulteration is a global concern, and the current study was plan to synthesize Selenium nanoparticles by green method using plant extract of garlic, Allium Sativum, and to characterize Selenium nanoparticles through different analytical techniques and to apply Selenium nanoparticles as fast and easy technique for the detection of salicylic acid in milk. The highly selective, sensitive, and quick interference green synthesis-based sensing of possible milk adulterants i.e., salicylic acid, has been reported here. Salicylic acid interacts with nanoparticles through strong bonding interactions, hence resulting in an interruption within the formation of selenium nanoparticles which is confirmed by UV-VIS spectroscopy, scanning electron microscopy, and x-ray diffraction. This interaction in the synthesis of nanoparticles resulted in transmittance wavelength that decrease with the increasing amount of salicylic acid, showing strong binding of selenium nanoparticles with adulterant, thereby permitting in-situ fast detection of salicylic acid from milk having a limit of detection at 10-3 mol and linear coefficient correlation of 0.9907. Conclusively, it can be draw that colloidal selenium could be synthesize successfully by garlic extract in order to serve as a probe for fast and cheap testing of milk adulteration.Keywords: adulteration, green synthesis, selenium nanoparticles, salicylic acid, aggregation
Procedia PDF Downloads 80856 Oat βeta Glucan Attenuates the Development of Atherosclerosis and Improves the Intestinal Barrier Function by Reducing Bacterial Endotoxin Translocation in APOE-/- MICE
Authors: Dalal Alghawas, Jetty Lee, Kaisa Poutanen, Hani El-Nezami
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Oat β-glucan a water soluble non starch linear polysaccharide has been approved as a cholesterol lowering agent by various food safety administrations and is commonly used to reduce the risk of heart disease. The molecular weight of oat β-glucan can vary depending on the extraction and fractionation methods. It is not clear whether the molecular weight has a significant impact at reducing the acceleration of atherosclerosis. The aim of this study was to investigate three different oat β-glucan fractionations on the development of atherosclerosis in vivo. With special focus on plaque stability and the intestinal barrier function. To test this, ApoE-/- female mice were fed a high fat diet supplemented with oat bran, high molecular weight (HMW) oat β-glucan fractionate and low molecular weight (LMW) oat β-glucan fractionate for 16 weeks. Atherosclerosis risk markers were measured in the plasma, heart and aortic tree. Plaque size was measured in the aortic root and aortic tree. ICAM-1, VCAM-1, E-Selectin, P-Selectin, protein levels were assessed from the aortic tree to determine plaque stability at 16 weeks. The expression of p22phox at the aortic root was evaluated to study the NADPH oxidase complex involved in nitric oxide bioavailability and vascular elasticity. The tight junction proteins E-cadherin and beta-catenin from western blot analyses were analysed as an intestinal barrier function test. Plasma LPS, intestinal D-lactate levels and hepatic FMO gene expression were carried out to confirm whether the compromised intestinal barrier lead to endotoxemia. The oat bran and HMW oat β-glucan diet groups were more effective than the LMW β-glucan diet group at reducing the plaque size and showed marked improvements in plaque stability. The intestinal barrier was compromised for all the experimental groups however the endotoxemia levels were higher in the LMW β-glucan diet group. The oat bran and HMW oat β-glucan diet groups were more effective at attenuating the development of atherosclerosis. Reasons for this could be due to the LMW oat β-glucan diet group’s low viscosity in the gut and the inability to block the reabsorption of cholesterol. Furthermore the low viscosity may allow more bacterial endotoxin translocation through the impaired intestinal barrier. In future food technologists should carefully consider how to incorporate LMW oat β-glucan as a health promoting food.Keywords: Atherosclerosis, beta glucan, endotoxemia, intestinal barrier function
Procedia PDF Downloads 418855 Speciation of Bacteria Isolated from Clinical Canine and Feline Urine Samples by Using ChromID CPS Elite Agar: A Preliminary Study
Authors: Delsy Salinas, Andreia Garcês, Augusto Silva, Paula Brilhante Simões
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Urinary tract infection (UTI) is a common disease affecting dogs and cats in both community and hospital environment. Bacteria is the most frequent agent isolated, fewer than 1% of infections are due to parasitic, fungal, or viral agents. Common symptoms and laboratory abnormalities includeabdominal pain, pyrexia, renomegaly, and neutrophilia with left shift. A rapid and precise identification of the bacterial agent is still a challenge in veterinarian laboratories. Therefore, this cross-sectional study aims to describe bacterial colony patterns of urine samples by using chromID™ CPS® EliteAgar (BioMérieux, France) from canine and feline specimens submitted to a veterinary laboratory in Portugal (INNO Veterinary Laboratory, Braga)from January to March2022. All urine samples were cultivated in CPS Elite Agar with calibrated 1 µL inoculating loop and incubated at 37ºC for 18-24h. Color,size, and shape (regular or irregular outline)were recorded for all samples. All colonies were classified as Gram-negative or Gram-positive bacteriausing Gram stain (PREVI® Color BioMérieux, France) and determined if they were pure colonies. Identification of bacteria species was performed using GP and GN cards inVitek 2® Compact(BioMérieux, France). A total of 256/1003 submitted urine samples presented bacterial growth, from which 172 isolates were included in this study. The sample’s population included 111 dogs (n=45 males and n=66 females) and 61 cats (n=35 males and n=26 females). The most frequent isolated bacteria wasEscherichia coli (44,7%), followed by Proteus mirabilis (13,4%). All Escherichia coli isolates presented red to burgundy colonies, a colony diameter between 2 to 6 mm, and regular or irregular outlines. Similarly, 100% of Proteus mirabilis isolates were dark yellow colonies with a diffuse pigment and the same size and shape as Escherichia coli. White and pink pale colonies where Staphylococcus species exclusively and S. pseudintermedius was the most frequent (8,2 %). Cian to blue colonies were mostly Enterococcusspp. (8,2%) and Streptococcus spp. (4,6%). Beige to brown colonies were Pseudomonas aeruginosa (2,9%) and Citrobacter spp. (1,2%).Klebsiella spp.,Serratia spp. and Enterobacter spp were green colonies. All Gram-positive isolates were 1 to 2 mm diameter long and had a regular outline, meanwhile, Gram-negative rods presented variable patterns. This results showed that theprevalence of E coli and P. mirabilis as uropathogenic agents follows the same trends in Europe as previously described in other studies. Both agents presented a particular color pattern in CPS Elite Agar to identify them without needing complementary tests. No other bacteria genus could be correlated strongly to a specific color pattern, and similar results have been observed instudies using human’s samples. Chromogenic media shows a great advantage for common urine bacteria isolation than traditional COS, McConkey, and CLEDAgar mediums in a routine context, especially when mixed fermentative Gram-negative agents grow simultaneously. In addition, CPS Elite Agar is versatile for Artificial Intelligent Reading Plates Systems. Routine veterinarian laboratories could use CPS Elite Agar for a rapid screening for bacteria identification,mainlyE coli and P.mirabilis, saving 6h to 10h of automatized identification.Keywords: cats, CPS elite agar, dogs, urine pathogens
Procedia PDF Downloads 102854 Probiotics as an Alternative to Antibiotic Use in Pig Production
Authors: Z. C. Dlamini, R. L. S. Langa, A. I. Okoh, O. A. Aiyegoro
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The indiscriminate usage of antibiotics in swine production have consequential outcomes; such as development of bacterial resistance to prophylactic antibiotics and possibility of antibiotic residues in animal products. The use of probiotics appears to be the most effective procedure with positive metabolic nutritional implications. The aim of this study was to investigate the efficacy of probiotic bacteria (Lactobacillus reuteri ZJ625, Lactobacillus reuteri VB4, Lactobacillus salivarius ZJ614 and Streptococcus salivarius NBRC13956) administered as direct-fed microorganisms in weaned piglets. 45 weaned piglets blocked by weight were dived into 5 treatments groups: diet with antibiotic, diet with no-antibiotic and no probiotic, and diet with probiotic and diet with combination of probiotics. Piglets performance was monitored during the trials. Faecal and Ileum samples were collected for microbial count analysis. Blood samples were collected from pigs at the end of the trial, for analysis of haematological, biochemical and IgG stimulation. The data was analysed by Split-Plot ANOVA using SAS statistically software (SAS 9.3) (2003). The difference was observed between treatments for daily weight and feed conversion ratio. No difference was observed in analysis of faecal samples in regards with bacterial counts, difference was observed in ileums samples with enteric bacteria colony forming unit being lower in P2 treatment group as compared with lactic acid and total bacteria. With exception of globulin and albumin, biochemistry blood parameters were not affected, likewise for haematology, only basophils and segmented neutrophils were differed by having higher concentration in NC treatment group as compared with other treatment groups. Moreover, in IgG stimulation analysis, difference was also observed, with P2 treatment group having high concentration of IgG in P2 treatment group as compared to other groups. The results of this study suggest that probiotics have a beneficial effect on growth performances, blood parameters and IgG stimulation of pigs, most effective when they are administered in synergy form. This means that it is most likely that these probiotics will offer a significant benefit in pig farming by reducing risk of morbidity and mortality and produce quality meat that is more affordable to poorer communities, and thereby enhance South African pig industry’s economy. In addition, these results indicate that there is still more research need to be done on probiotics in regards with, i.e. dosage, shelf life and mechanism of action.Keywords: antibiotics, biochemistry, haematology, IgG-stimulation, microbial count, probiotics
Procedia PDF Downloads 299853 Mathematical Modeling of the Fouling Phenomenon in Ultrafiltration of Latex Effluent
Authors: Amira Abdelrasoul, Huu Doan, Ali Lohi
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An efficient and well-planned ultrafiltration process is becoming a necessity for monetary returns in the industrial settings. The aim of the present study was to develop a mathematical model for an accurate prediction of ultrafiltration membrane fouling of latex effluent applied to homogeneous and heterogeneous membranes with uniform and non-uniform pore sizes, respectively. The models were also developed for an accurate prediction of power consumption that can handle the large-scale purposes. The model incorporated the fouling attachments as well as chemical and physical factors in membrane fouling for accurate prediction and scale-up application. Both Polycarbonate and Polysulfone flat membranes, with pore sizes of 0.05 µm and a molecular weight cut-off of 60,000, respectively, were used under a constant feed flow rate and a cross-flow mode in ultrafiltration of the simulated paint effluent. Furthermore, hydrophilic ultrafilic and hydrophobic PVDF membranes with MWCO of 100,000 were used to test the reliability of the models. Monodisperse particles of 50 nm and 100 nm in diameter, and a latex effluent with a wide range of particle size distributions were utilized to validate the models. The aggregation and the sphericity of the particles indicated a significant effect on membrane fouling.Keywords: membrane fouling, mathematical modeling, power consumption, attachments, ultrafiltration
Procedia PDF Downloads 469852 The Impact of Efflux Pump Inhibitor on the Activity of Benzosiloxaboroles and Benzoxadiboroles against Gram-Negative Rods
Authors: Agnieszka E. Laudy, Karolina Stępien, Sergiusz Lulinski, Krzysztof Durka, Stefan Tyski
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1,3-dihydro-1-hydroxy-2,1-benzoxaborole and its derivatives are a particularly interesting group of synthetic agents and were successfully employed in supramolecular chemistry medicine. The first important compounds, 5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole and 5-chloro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole were identified as potent antifungal agents. In contrast, (S)-3-(aminomethyl)-7-(3-hydroxypropoxy)-1-hydroxy-1,3-dihydro-2,1-benzoxaborole hydrochloride is in the second phase of clinical trials as a drug for the treatment of Gram-negative bacterial infections of the Enterobacteriaceae family and Pseudomonas aeruginosa. Equally important and difficult task is to search for compounds active against Gram-negative bacilli, which have multi-drug-resistance efflux pumps actively removing many of the antibiotics from bacterial cells. We have examined whether halogen-substituted benzoxaborole-based derivatives and their analogues possess antibacterial activity and are substrates for multi-drug-resistance efflux pumps. The antibacterial activity of 1,3-dihydro-3-hydroxy-1,1-dimethyl-1,2,3-benzosiloxaborole and 10 halogen-substituted its derivatives, as well as 1,2-phenylenediboronic acid and 3 synthesised fluoro-substituted its analogs, were evaluated. The activity against the reference strains of Gram-positive (n=5) and Gram-negative bacteria (n=10) was screened by the disc-diffusion test (0.4 mg of tested compounds was applied onto paper disc). The minimal inhibitory concentration values and the minimal bactericidal concentration values were estimated according to The Clinical and Laboratory Standards Institute and The European Committee on Antimicrobial Susceptibility Testing recommendations. During the minimal inhibitory concentration values determination with or without phenylalanine-arginine beta-naphthylamide (50 mg/L) efflux pump inhibitor, the concentrations of tested compounds ranged 0.39-400 mg/L in the broth medium supplemented with 1 mM magnesium sulfate. Generally, the studied benzosiloxaboroles and benzoxadiboroles showed a higher activity against Gram-positive cocci than against Gram-negative rods. Moreover, benzosiloxaboroles have the higher activity than benzoxadiboroles compounds. In this study, we demonstrated that substitution (mono-, di- or tetra-) of 1,3-dihydro-3-hydroxy-1,1-dimethyl-1,2,3-benzosiloxaborole with halogen groups resulted in an increase in antimicrobial activity as compared to the parent substance. Interestingly, the 6,7-dichloro-substituted parent substance was found to be the most potent against Gram-positive cocci: Staphylococcus sp. (minimal inhibitory concentration 6.25 mg/L) and Enterococcus sp. (minimal inhibitory concentration 25 mg/L). On the other hand, mono- and dichloro-substituted compounds were the most actively removed by efflux pumps present in Gram-negative bacteria mainly from Enterobacteriaceae family. In the presence of efflux pump inhibitor the minimal inhibitory concentration values of chloro-substituted benzosiloxaboroles decreased from 400 mg/L to 3.12 mg/L. Of note, the highest increase in bacterial susceptibility to tested compounds in the presence of phenylalanine-arginine beta-naphthylamide was observed for 6-chloro-, 6,7-dichloro- and 6,7-difluoro-substituted benzosiloxaboroles. In the case of Escherichia coli, Enterobacter cloacae and P. aeruginosa strains at least a 32-fold decrease in the minimal inhibitory concentration values of these agents were observed. These data demonstrate structure-activity relationships of the tested derivatives and highlight the need for further search for benzoxaboroles and related compounds with significant antimicrobial properties. Moreover, the influence of phenylalanine-arginine beta-naphthylamide on the susceptibility of Gram-negative rods to studied benzosiloxaboroles indicate that some tested agents are substrates for efflux pumps in Gram-negative rods.Keywords: antibacterial activity, benzosiloxaboroles, efflux pumps, phenylalanine-arginine beta-naphthylamide
Procedia PDF Downloads 269851 Ayurvastra: A Study on the Ancient Indian Textile for Healing
Authors: Reena Aggarwal
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The use of textile chemicals in the various pre and post-textile manufacturing processes has made the textile industry conscious of its negative contribution to environmental pollution. Popular environmentally friendly fibers such as recycled polyester and organic cotton have been now increasingly used by fabrics and apparel manufacturers. However, after these textiles or the finished apparel are manufactured, they have to be dyed in the same chemical dyes that are harmful and toxic to the environment. Dyeing is a major area of concern for the environment as well as for people who have chemical sensitivities as it may cause nausea, breathing difficulties, seizures, etc. Ayurvastra or herbal medical textiles are one step ahead of the organic lifestyle, which supports the core concept of holistic well-being and also eliminates the impact of harmful chemicals and pesticides. There is a wide range of herbs that can be used not only for dyeing but also for providing medicinal properties to the textiles like antibacterial, antifungal, antiseptic, antidepressant and for treating insomnia, skin diseases, etc. The concept of herbal dyeing of fabric is to manifest herbal essence in every aspect of clothing, i.e., from production to end-use, additionally to eliminate the impact of harmful chemical dyes and chemicals which are known to result in problems like skin rashes, headache, trouble concentrating, nausea, diarrhea, fatigue, muscle and joint pain, dizziness, difficulty breathing, irregular heartbeat and seizures. Herbal dyeing or finishing on textiles will give an extra edge to the textiles as it adds an extra function to the fabric. The herbal extracts can be applied to the textiles by a simple process like the pad dry cure method and mainly acts on the human body through the skin for aiding in the treatment of disease or managing the medical condition through its herbal properties. This paper, therefore, delves into producing Ayurvastra, which is a perfect amalgamation of cloth and wellness. The aim of the paper is to design and create herbal disposable and non-disposable medical textile products acting mainly topically (through the skin) for providing medicinal properties/managing medical conditions. Keeping that in mind, a range of antifungal socks and antibacterial napkins treated with turmeric and aloe vera were developed, which are recommended for the treatment of fungal and bacterial infections, respectively. Both Herbal Antifungal socks and Antibacterial napkins have proved to be efficient enough in managing and treating fungal and bacterial infections of the skin, respectively.Keywords: ayurvastra, ayurveda, herbal, pandemic, sustainable
Procedia PDF Downloads 130850 AG Loaded WO3 Nanoplates for Photocatalytic Degradation of Sulfanilamide and Bacterial Removal under Visible Light
Authors: W. Y. Zhu, X. L. Yan, Y. Zhou
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Sulfonamides (SAs) are extensively used antibiotics; photocatalysis is an effective, way to remove the SAs from water driven by solar energy. Here we used WO3 nanoplates and their Ag heterogeneous as photocatalysts to investigate their photodegradation efficiency against sulfanilamide (SAM) which is the precursor of SAs. Results showed that WO3/Ag composites performed much better than pure WO3 where the highest removal rate was 96.2% can be achieved under visible light irradiation. Ag as excellent antibacterial agent also endows certain antibacterial efficiency to WO3, and 100% removal efficiency could be achieved in 2 h under visible light irradiation for all WO3/Ag composites. Generally, WO3/Ag composites are very effective photocatalysts with potentials in practical applications which mainly use cheap, clean and green solar energy as energy source.Keywords: antibacterial, photocatalysis, semiconductor, sulfanilamide
Procedia PDF Downloads 357849 Explainable Graph Attention Networks
Authors: David Pham, Yongfeng Zhang
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Graphs are an important structure for data storage and computation. Recent years have seen the success of deep learning on graphs such as Graph Neural Networks (GNN) on various data mining and machine learning tasks. However, most of the deep learning models on graphs cannot easily explain their predictions and are thus often labelled as “black boxes.” For example, Graph Attention Network (GAT) is a frequently used GNN architecture, which adopts an attention mechanism to carefully select the neighborhood nodes for message passing and aggregation. However, it is difficult to explain why certain neighbors are selected while others are not and how the selected neighbors contribute to the final classification result. In this paper, we present a graph learning model called Explainable Graph Attention Network (XGAT), which integrates graph attention modeling and explainability. We use a single model to target both the accuracy and explainability of problem spaces and show that in the context of graph attention modeling, we can design a unified neighborhood selection strategy that selects appropriate neighbor nodes for both better accuracy and enhanced explainability. To justify this, we conduct extensive experiments to better understand the behavior of our model under different conditions and show an increase in both accuracy and explainability.Keywords: explainable AI, graph attention network, graph neural network, node classification
Procedia PDF Downloads 197848 A Theoretical to Conceptual Paper: The Use of Phosphodiesterase Inhibitors, Endothelin Receptor Antagonists and/or Prostacyclin Analogs in Acute Pulmonary Embolism
Authors: Ryan M. Monti, Bijal Mehta
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In cases of massive pulmonary embolism, defined as acute pulmonary embolism presenting with systemic hypotension or right ventricular dysfunction and impending failure, there is indication that unconventional therapies, such as phosphodiesterase inhibitors, endothelin receptor antagonists, and/or prostacyclin analogs may decrease the morbidity and mortality. Based on the premise that dilating the pulmonary artery will decrease the pulmonary vascular pressure, while simultaneously decreasing the aggregation of platelets, it can be hypothesized that increased blood flow through the pulmonary artery will decrease right heart strain and subsequent morbidity and mortality. While this theory has yet to be formally studied, the recommendations for treating massive pulmonary embolism with phosphodiesterase inhibitors, endothelin receptor antagonists, and/or prostacyclin analogs in conjunction with the current standards of care in massive pulmonary embolism should be formally studied. In particular, patients with massive PE who are unable to undergo thrombolysis/surgical intervention may be the ideal population to study the use of these treatments to determine any decrease in mortality and morbidity (short term and long term).Keywords: acute pulmonary thromboembolism, treatment of pulmonary embolism, use of phosphodiesterase inhibitors, endothelin receptor antagonists, prostacyclin analogs in PE
Procedia PDF Downloads 224847 The Modeling of Viscous Microenvironment for the Coupled Enzyme System of Bioluminescence Bacteria
Authors: Irina E. Sukovataya, Oleg S. Sutormin, Valentina A. Kratasyuk
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Effect of viscosity of media on kinetic parameters of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase was investigated with addition of organic solvents (glycerol and sucrose), because bioluminescent enzyme systems based on bacterial luciferases offer a unique and general tool for analysis of the many analytes and enzymes in the environment, research, and clinical laboratories and other fields. The possibility of stabilization and increase of activity of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase activity in vicious aqueous-organic mixtures have been shown.Keywords: coupled enzyme system of bioluminescence bacteria NAD(P)H:FMN-oxidoreductase–luciferase, glycerol, stabilization of enzymes, sucrose
Procedia PDF Downloads 393846 Wireless Sensor Network Energy Efficient and QoS-Aware MAC Protocols: A Survey
Authors: Bashir Abdu Muzakkari, Mohamad Afendee Mohamad, Mohd Fadzil Abdul Kadir
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Wireless Sensor Networks (WSNs) is an aggregation of several tiny, low-cost sensor nodes, spatially distributed to monitor physical or environmental status. WSN is constantly changing because of the rapid technological advancements in sensor elements such as radio, battery and operating systems. The Medium Access Control (MAC) protocols remain very vital in the WSN because of its role in coordinating communication amongst the sensors. Other than battery consumption, packet collision, network lifetime and latency are factors that largely depend on WSN MAC protocol and these factors have been widely treated in recent days. In this paper, we survey some latest proposed WSN Contention-based, Scheduling-based and Hybrid MAC protocols while presenting an examination, correlation of advantages and limitations of each protocol. Concentration is directed towards investigating the treatment of Quality of Service (QoS) performance metrics within these particular protocols. The result shows that majority of the protocols leaned towards energy conservation. We, therefore, believe that other performance metrics of guaranteed QoS such as latency, throughput, packet loss, network and bandwidth availability may play a critical role in the design of future MAC protocols for WSNs.Keywords: WSN, QoS, energy consumption, MAC protocol
Procedia PDF Downloads 398845 Correlation Mapping for Measuring Platelet Adhesion
Authors: Eunseop Yeom
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Platelets can be activated by the surrounding blood flows where a blood vessel is narrowed as a result of atherosclerosis. Numerous studies have been conducted to identify the relation between platelets activation and thrombus formation. To measure platelet adhesion, this study proposes an image analysis technique. Blood samples are delivered in the microfluidic channel, and then platelets are activated by a stenotic micro-channel with 90% severity. By applying proposed correlation mapping, which visualizes decorrelation of the streaming blood flow, the area of adhered platelets (APlatelet) was estimated without labeling platelets. In order to evaluate the performance of correlation mapping on the detection of platelet adhesion, the effect of tile size was investigated by calculating 2D correlation coefficients with binary images obtained by manual labeling and the correlation mapping method with different sizes of the square tile ranging from 3 to 50 pixels. The maximum 2D correlation coefficient is observed with the optimum tile size of 5×5 pixels. As the area of the platelet adhesion increases, the platelets plug the channel and there is only a small amount of blood flows. This image analysis could provide new insights for better understanding of the interactions between platelet aggregation and blood flows in various physiological conditions.Keywords: platelet activation, correlation coefficient, image analysis, shear rate
Procedia PDF Downloads 335844 Antagonistic Potential of Epiphytic Bacteria Isolated in Kazakhstan against Erwinia amylovora, the Causal Agent of Fire Blight
Authors: Assel E. Molzhigitova, Amankeldi K. Sadanov, Elvira T. Ismailova, Kulyash A. Iskandarova, Olga N. Shemshura, Ainur I. Seitbattalova
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Fire blight is a very harmful for commercial apple and pear production quarantine bacterial disease. To date, several different methods have been proposed for disease control, including the use of copperbased preparations and antibiotics, which are not always reliable or effective. The use of bacteria as biocontrol agents is one of the most promising and eco-friendly alternative methods. Bacteria with protective activity against the causal agent of fire blight are often present among the epiphytic microorganisms of the phyllosphere of host plants. Therefore, the main objective of our study was screening of local epiphytic bacteria as possible antagonists against Erwinia amylovora, the causal agent of fire blight. Samples of infected organs of apple and pear trees (shoots, leaves, fruits) were collected from the industrial horticulture areas in various agro-ecological zones of Kazakhstan. Epiphytic microorganisms were isolated by standard and modified methods on specific nutrient media. The primary screening of selected microorganisms under laboratory conditions to determine the ability to suppress the growth of Erwinia amylovora was performed by agar-diffusion-test. Among 142 bacteria isolated from the fire blight host plants, 5 isolates, belonging to the genera Bacillus, Lactobacillus, Pseudomonas, Paenibacillus and Pantoea showed higher antagonistic activity against the pathogen. The diameters of inhibition zone have been depended on the species and ranged from 10 mm to 48 mm. The maximum diameter of inhibition zone (48 mm) was exhibited by B. amyloliquefaciens. Less inhibitory effect was showed by Pantoea agglomerans PA1 (19 mm). The study of inhibitory effect of Lactobacillus species against E. amylovora showed that among 7 isolates tested only one (Lactobacillus plantarum 17M) demonstrated inhibitory zone (30 mm). In summary, this study was devoted to detect the beneficial epiphytic bacteria from plants organs of pear and apple trees due to fire blight control in Kazakhstan. Results obtained from the in vitro experiments showed that the most efficient bacterial isolates are Lactobacillus plantarum 17M, Bacillus amyloliquefaciens MB40, and Pantoea agglomerans PA1. These antagonists are suitable for development as biocontrol agents for fire blight control. Their efficacies will be evaluated additionally, in biological tests under in vitro and field conditions during our further study.Keywords: antagonists, epiphytic bacteria, Erwinia amylovora, fire blight
Procedia PDF Downloads 166843 Bioaccessibility of Vitamin A Nanoemulsion: Influence of Carrier Oil and Surfactant Concentration
Authors: R. N. Astya, E. S. Nugraha, S. P. Nurheni, Hoerudin
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Vitamin A deficiency remains to be among the major malnutrition problems in Indonesia. Vitamin A is a fat-soluble vitamin which renders it difficult to be fortified in water-based foods and beverages. Furthermore, its low solubility and stability in aqueous system may limit its bioaccessibility in the gastrointestinal tract. Nanoemulsification of vitamin A may solve these problems. The objective of this study was to investigate bioaccessibility of vitamin A (retinyl palmitate/RP) nanoemulsion as influenced by two types of carrier oil (Virgin Coconut Oil/VCO and corn oil/CO) and surfactant concentrations (polysorbate 20/Tween 20 3% and 6%). Oil in water (o/w) nanoemulsions of vitamin A was produced through a combination of high shear-high pressure homogenization technique. The results showed that RP-VCO nanoemulsions were 121.62 nm (3%) and 115.40 (6%) nm in particle size, whereas RP-CO nanoemulsions were 154.72 nm (3%) and 134.00 nm (6%) in particle size. As for VCO nanoemulsions, the bioaccessibility of vitamin A was shown to be 89.84% and 55.32%, respectively. On the other hand, CO nanoemulsions produced vitamin A bioaccessibility of 53.66% and 44.85%, respectively. In general, VCO nanoemulsions showed better bioaccessibility of vitamin A than CO nanoemulsions. In this study, RP-VCO nanoemulsion with 3% Tween 20 had the highest ζ-potential value (-26.5 mV) and produced the highest bioaccessibility of vitamin A (89.84%, P<0.05). Additionally, the vitamin A nanoemulsion was stable even for after a week of freeze and thaw treatment. Following the freeze and thaw treatment, the vitamin A nanoemulsion showed good stability without aggregation and separation. These results would be useful for designing effective vitamin A delivery systems for food and beverage applications.Keywords: bioaccessibility, carrier oil, surfactant, vitamin A nanoemulsion
Procedia PDF Downloads 269842 Lab Activities for Introducing Nanoscience to Teachers and Students
Authors: Riam Abu-Much, Muhamad Hugerat
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Nanoscience has become one of the main science fields in the world; its importance is reflected in both society and industry; therefore, it is very important to intensify educational programs among teachers and students that aim to introduce "Nano Concepts" to them. Two different lab activities were developed for demonstrating the importance of nanoscale materials using unique points of view. In the first, electrical conductive films made of silver nanoparticles were fabricated. The silver nanoparticles were protected against aggregation using electrical conductive polypyrrole, which acts also as conductive bridge between them. The experiments show a simpler way for fabricating conductive thin film than the much more complicated and costly conventional method. In the second part, the participants could produce emulsions of liposome structures using Phosphatidylcholine as a surfactant, and following by minimizing the size of it from micro-scale to nanometer scale (400 nm), using simple apparatus called Mini-Extruder, in that way the participants could realize the change in solution transparency, and the effect of Tyndall when the size of the liposomes is reduced. Freshmen students from the Academic Arab College for Education in Haifa, Israel, who are studying to become science teachers, participated in this lab activity as part of the course "Chemistry in the Lab". These experiments are appropriate for teachers, high school and college students.Keywords: case study, colloid, emulsion, liposome, surfactant
Procedia PDF Downloads 201841 Investigating the Essentiality of Oxazolidinones in Resistance-Proof Drug Combinations in Mycobacterium tuberculosis Selected under in vitro Conditions
Authors: Gail Louw, Helena Boshoff, Taeksun Song, Clifton Barry
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Drug resistance in Mycobacterium tuberculosis is primarily attributed to mutations in target genes. These mutations incur a fitness cost and result in bacterial generations that are less fit, which subsequently acquire compensatory mutations to restore fitness. We hypothesize that mutations in specific drug target genes influence bacterial metabolism and cellular function, which affects its ability to develop subsequent resistance to additional agents. We aim to determine whether the sequential acquisition of drug resistance and specific mutations in a well-defined clinical M. tuberculosis strain promotes or limits the development of additional resistance. In vitro mutants resistant to pretomanid, linezolid, moxifloxacin, rifampicin and kanamycin were generated from a pan-susceptible clinical strain from the Beijing lineage. The resistant phenotypes to the anti-TB agents were confirmed by the broth microdilution assay and genetic mutations were identified by targeted gene sequencing. Growth of mono-resistant mutants was done in enriched medium for 14 days to assess in vitro fitness. Double resistant mutants were generated against anti-TB drug combinations at concentrations 5x and 10x the minimum inhibitory concentration. Subsequently, mutation frequencies for these anti-TB drugs in the different mono-resistant backgrounds were determined. The initial level of resistance and the mutation frequencies observed for the mono-resistant mutants were comparable to those previously reported. Targeted gene sequencing revealed the presence of known and clinically relevant mutations in the mutants resistant to linezolid, rifampicin, kanamycin and moxifloxacin. Significant growth defects were observed for mutants grown under in vitro conditions compared to the sensitive progenitor. Mutation frequencies determination in the mono-resistant mutants revealed a significant increase in mutation frequency against rifampicin and kanamycin, but a significant decrease in mutation frequency against linezolid and sutezolid. This suggests that these mono-resistant mutants are more prone to develop resistance to rifampicin and kanamycin, but less prone to develop resistance against linezolid and sutezolid. Even though kanamycin and linezolid both inhibit protein synthesis, these compounds target different subunits of the ribosome, thereby leading to different outcomes in terms of fitness in the mutants with impaired cellular function. These observations showed that oxazolidinone treatment is instrumental in limiting the development of multi-drug resistance in M. tuberculosis in vitro.Keywords: oxazolidinones, mutations, resistance, tuberculosis
Procedia PDF Downloads 160840 Preparation and in vitro Bactericidal and Fungicidal Efficiency of NanoSilver/Methylcellulose Hydrogel
Authors: A. Panacek, M. Kilianova, R. Prucek, V. Husickova, R. Vecerova, M. Kolar, L. Kvitek, R. Zboril
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In this work we describe the preparation of NanoSilver/methylcellulose hydrogel containing silver nanoparticles (NPs) for topical bactericidal applications. Highly concentrated dispersion of silver NPs as high as of 5g/L of silver with diameter of 10nm was prepared by reduction of AgNO3 via strong reducing agent NaBH4. Silver NPs were stabilized by addition of sodium polyacrylate in order to prevent their aggregation at such high concentration. This way synthesized silver NPs were subsequently incorporated into methylcellulose suspension at elevated temperature resulting in formation of NanoSilver/methylcellulose hydrogel when temperature cooled down to laboratory conditions. In vitro antibacterial activity assay proved high bactericidal and fungicidal efficiency of silver NPs alone in the form of dispersion as well as in the form of hydrogel against broad spectrum of bacteria and yeasts including highly multiresistant strains such as methicillin-resistant Staphylococcus aureus. A very low concentrations of silver as low as 0.84mg/L Ag in as-prepared dispersion gave antibacterial performance. NanoSilver/methylcellulose hydrogel showed antibacterial action at the lowest used silver concentration equal to 25mg/L. Such prepared NanoSilver/methylcellulose hydrogel represent promising topical antimicrobial formulation for treatment of burns and wounds.Keywords: antimicrobial, burn, hydrogel, silver NPs
Procedia PDF Downloads 449839 TiO2/Clay Minerals (Palygorskite/Halloysite) Nanocomposite Coatings for Water Disinfection
Authors: Dionisios Panagiotaras, Dimitrios Papoulis, Elias Stathatos
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Microfibrous palygorskite and tubular halloysite clay mineral combined with nanocrystalline TiO2 are incorporating in the preparation of nanocomposite films on glass substrates via sol-gel route at 450 °C. The synthesis is employing nonionic surfactant molecule as pore directing agent along with acetic acid-based sol-gel route without addition of water molecules. Drying and thermal treatment of composite films ensure elimination of organic material lead to the formation of TiO2 nanoparticles homogeneously distributed on the palygorskite or halloysite surfaces. Nanocomposite films without cracks of active anatase crystal phase on palygorskite and halloysite surfaces are characterized by microscopy techniques, UV-Vis spectroscopy, and porosimetry methods in order to examine their structural properties. The composite palygorskite-TiO2 and halloysite-TiO2 films with variable quantities of palygorskite and halloysite were tested as photocatalysts in the photo-oxidation of Basic Blue 41 azo dye in water. These nanocomposite films proved to be most promising photocatalysts and highly effective to dye’s decoloration in spite of small amount of palygorskite -TiO2 or halloysite- TiO2 catalyst immobilized onto glass substrates mainly due to the high surface area and uniform distribution of TiO2 on clay minerals avoiding aggregation.Keywords: halloysite, palygorskite, photocatalysis, titanium dioxide
Procedia PDF Downloads 313838 Fabrication of Nanostructured Arrays Using Si-Containing Block Copolymer and Dually Responsive Photoresist
Authors: Kyoungok Jung, Chang Hong Bak, Gyeong Cheon Jo, Jin-Baek Kim
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Nanostructured arrays have drawn extensive attention because of their unique properties resulting from nanoscale features. However, it is difficult to achieve uniform and freestanding 1D nanostrcutures over a large area. Here, a simple and novel method was developed for fabrication of universal nanoporous templates for high-density nanostructure arrays, by combining self-assembly of a Si-containing block copolymer with a bilayer lithography system. We introduced a dually responsive photoresist bottom layer into which the nanopatterns of block copolymer are transferred by oxygen reactive ion etching. Because the dually responsive layer becomes cross-linked by heating, it can be used as a hard template during the etching process. It becomes soluble again by chain scission upon exposure to light. Therefore, it can be easily removed by the lift-off process. The template was applicable to the various conducting substrates due to the compatibility of the photoresist with a wide range of substrates and was used in electrodeposition for well-aligned and high-density inorganic and organic nanoarrays. We successfully obtained vertically aligned and highly ordered gold nanorods and polypyrrole dots on the substrate without aggregation, and these arrays did not collapse after removing the dually responsive templates by the simple lift-off process.Keywords: block copolymer, dually responsive, nanostructure, photoresist
Procedia PDF Downloads 256837 Sustainable Development: Evaluation of an Urban Neighborhood
Authors: Harith Mohammed Benbouali
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The concept of sustainable development is becoming increasingly important in our society. The efforts of specialized agencies, cleverly portrayed in the media, allow a widespread environmental awareness. Far from the old environmental movement in the backward-looking nostalgia, the environment is combined with today's progress. Many areas now include these concerns in their efforts, this in order to try to reduce the negative impact of human activities on the environment. The quantitative dimension of development has given way to the quality aspect. However, this feature is not common, and the initial target was abandoned in favor of economic considerations. Specialists in the field of building and construction have constantly sought to further integrate the environmental dimension, creating a seal of high environmental quality buildings. The pursuit of well-being of neighborhood residents and the quality of buildings are also a hot topic in planning. Quality of life is considered so on, since financial concerns dominate to the detriment of the environment and the welfare of the occupants. This work concerns the development of an analytical method based on multiple indicators of objectives across the district. The quantification of indicators related to objectives allows the construction professional, the developer or the community, to quantify and compare different alternatives for development of a neighborhood. This quantification is based on the use of simulation tools and a multi-criteria aggregation.Keywords: sustainable development, environment, district, indicators, multi-criteria analysis, evaluation
Procedia PDF Downloads 311836 Inhibitory Effects of PPARγ Ligand, KR-62980, on Collagen-Stimulated Platelet Activation
Authors: Su Bin Wang, Jin Hee Ahn, Tong-Shin Chang
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The peroxisome proliferator-activated receptors (PPARs) are member of nuclear receptor superfamily that act as a ligand-activated transcription factors. Although platelets lack a nucleus, previous studies have shown that PPARγ agonists, rosiglitazone, inhibited platelet activation induced by collagen. In this study, we investigated the inhibitory effects of KR-62980, a newly synthesized PPARγ agonist, on collagen receptor-stimulated platelet activation. The specific tyrosine phosphorylations of key components (Syk, Vav1, Btk and PLCγ2) for collagen receptor signaling pathways were suppressed by KR-62980. KR-62980 also attenuated downstream responses including cytosolic calcium elevation, P-selectin surface exposure, and integrin αIIbβ3 activation. PPARγ was found to associate with multiple proteins within the LAT signaling complex in collagen-stimulated platelets. This association was prevented by KR-62980, indicating a potential mechanism for PPARγ function in collagen-stimulated platelet activation. Furthermore, KR-62980 inhibited platelet aggregation and adhesion in response to collagen in vitro and prolonged in vivo thrombotic response in carotid arteries of mice. Collectively, these data suggest that KR-62980 inhibits collagen-stimulated platelet activation and thrombus formation through modulating the collagen receptor signaling pathways.Keywords: KR-62980, PPARγ, antiplatelet, thrombosis
Procedia PDF Downloads 331