Search results for: microbial pretreatment
Commenced in January 2007
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Edition: International
Paper Count: 1185

Search results for: microbial pretreatment

375 Bioflocculation Using the Purified Wild Strain of P. aeruginosa Culture in Wastewater Treatment

Authors: Mohammad Hajjartabar, Tahereh Kermani Ranjbar

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P. aeruginosa EF2 was isolated and identified from human infection sources before in our previous study. The present study was performed to determine the characteristics and activity role of bioflocculant produced by the bacterium in flocculation of the wastewater active sludge treatment. The bacterium was inoculated and then was grown in an orbital shaker at 250 rpm for 5 days at 35 °C under TSB and peptone water media. After incubation period, culture broths of the bacterial strain was collected and washed. The concentration of the bacteria was adjusted. For the extraction of the bacterial bioflocculant, culture was centrifuged at 6000 rpm for 20 min at 4 °C to remove bacterial cells. Supernatant was decanted and pellet containing bioflocculant was dried at 105 °C to a constant weight according to APHA, 2005. The chemical composition of the extracted bioflocculant from the bacterial sample was then analyzed. Wastewater active sludge sample obtained from aeration tank from one of wastewater treatment plants in Tehran, was first mixed thoroughly. After addition of bioflocculant, improvements in floc density were observed with an increase in bioflocculant. The results of this study strongly suggested that the extracted bioflucculant played a significant role in flocculation of the wastewater sample. The use of wild bacteria and nutrient regulation techniques instead of genetic manipulation opens wide investigation area in the future to improve wastewater treatment processes. Also this may put a new path in front of us to attain and improve the more effective bioflocculant using the purified microbial culture in wastewater treatment.

Keywords: wastewater treatment, P. aeruginosa, sludge treatment

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374 Retrospective Study of Bronchial Secretions Cultures Carried out in the Microbiology Department of General Hospital of Ioannina in 2017

Authors: S. Mantzoukis, M. Gerasimou, P. Christodoulou, N. Varsamis, G. Kolliopoulou, N. Zotos

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Purpose: Patients in Intensive Care Units (ICU) are exposed to a different spectrum of microorganisms relative to the hospital. Due to the fact that the majority of these patients are intubated, bronchial secretions should be examined. Material and Method: Bronchial secretions should be taken with care so as not to be mixed with sputum or saliva. The bronchial secretions are placed in a sterile container and then inoculated into blood, Mac Conkey No2, Chocolate, Mueller Hinton, Chapman and Saboureaud agar. After this period, if any number of microbial colonies are detected, gram staining is performed and then the isolated organisms are identified by biochemical techniques in the automated Microscan system (Siemens) followed by a sensitivity test in the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by a Kirby Bauer test. Results: In 2017 the Laboratory of Microbiology received 365 samples of bronchial secretions from the Intensive Care Unit. 237 were found positive. S. epidermidis was identified in 1 specimen, A. baumannii in 60, K. pneumoniae in 42, P. aeruginosa in 50, C. albicans in 40, P. mirabilis in 4, E. coli in 4, S. maltophilia in 6, S. marcescens in 6, S. aureus in 12, S. pneumoniae in 1, S. haemolyticus in 4, P. fluorescens in 1, E. aerogenes in 1, E. cloacae in 5. Conclusions: The majority of ICU patients appear to be a fertile ground for the development of infections. The nature of the findings suggests that a significant part of the bacteria found comes from the unit (nosocomial infection).

Keywords: bronchial secretions, cultures, infections, intensive care units

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373 An Evaluative Microbiological Risk Assessment of Drinking Water Supply in the Carpathian Region: Identification of Occurrent Hazardous Bacteria with Quantitative Microbial Risk Assessment Method

Authors: Anikó Kaluzsa

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The article's author aims to introduce and analyze those microbiological safety hazards which indicate the presence of secondary contamination in the water supply system. Since drinking water belongs to primary foods and is the basic condition of life, special attention should be paid on its quality. There are such indicators among the microbiological features can be found in water, which are clear evidence of the presence of water contamination, and based on this there is no need to perform other diagnostics, because they prove properly the contamination of the given water supply section. Laboratory analysis can help - both technologically and temporally – to identify contamination, but it does matter how long takes the removal and if the disinfection process takes place in time. The identification of the factors that often occur in the same places or the chance of their occurrence is greater than the average, facilitates our work. The pathogen microbiological risk assessment by the help of several features determines the most likely occurring microbiological features in the Carpathian basin. From among all the microbiological indicators, that are recommended targets for routine inspection by the World Health Organization, there is a paramount importance of the appearance of Escherichia coli in the water network, as its presence indicates the potential ubietiy of enteric pathogens or other contaminants in the water network. In addition, the author presents the steps of microbiological risk assessment analyzing those pathogenic micro-organisms registered to be the most critical.

Keywords: drinking water, E. coli, microbiological indicators, risk assessment, water safety plan

Procedia PDF Downloads 305
372 Bacteriological Culture Methods and its Uses in Clinical Pathology

Authors: Prachi Choudhary, Jai Gopal Sharma

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Microbial cultures determine the type of organism, its abundance in the tested sample, or both. It is one of the primary diagnostic methods of microbiology. It is used to determine the cause of infectious disease by letting the agent multiply in a predetermined medium. Different bacteria produce colonies that may be very distinct from the bacterial species that produced them. To culture any pathogen or microorganism, we should first know about the types of media used in microbiology for culturing. Sometimes sub culturing is also done in various microorganisms if some mixed growth is seen in culture. Nearly 3 types of culture media based on consistency – solid, semi-solid, and liquid (broth) media; are further explained in the report. Then, The Five I's approach is a method for locating, growing, observing, and characterizing microorganisms, including inoculation and incubation. Isolation, inspection, and identification. For identification of bacteria, we have to culture the sample like urine, sputum, blood, etc., on suitable media; there are different methods of culturing the bacteria or microbe like pour plate method, streak plate method, swabbing by needle, pipetting, inoculation by loop, spreading by spreader, etc. After this, we see the bacterial growth after incubation of 24 hours, then according to the growth of bacteria antibiotics susceptibility test is conducted; this is done for sensitive antibiotics or resistance to that bacteria, and also for knowing the name of bacteria. Various methods like the dilution method, disk diffusion method, E test, etc., do antibiotics susceptibility tests. After that, various medicines are provided to the patients according to antibiotic sensitivity and resistance.

Keywords: inoculation, incubation, isolation, antibiotics suspectibility test, characterizing

Procedia PDF Downloads 48
371 Dynamic Analysis of the Heat Transfer in the Magnetically Assisted Reactor

Authors: Tomasz Borowski, Dawid Sołoducha, Rafał Rakoczy, Marian Kordas

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The application of magnetic field is essential for a wide range of technologies or processes (i.e., magnetic hyperthermia, bioprocessing). From the practical point of view, bioprocess control is often limited to the regulation of temperature at constant values favourable to microbial growth. The main aim of this study is to determine the effect of various types of electromagnetic fields (i.e., static or alternating) on the heat transfer in a self-designed magnetically assisted reactor. The experimental set-up is equipped with a measuring instrument which controlled the temperature of the liquid inside the container and supervised the real-time acquisition of all the experimental data coming from the sensors. Temperature signals are also sampled from generator of magnetic field. The obtained temperature profiles were mathematically described and analyzed. The parameters characterizing the response to a step input of a first-order dynamic system were obtained and discussed. For example, the higher values of the time constant means slow signal (in this case, temperature) increase. After the period equal to about five-time constants, the sample temperature nearly reached the asymptotic value. This dynamical analysis allowed us to understand the heating effect under the action of various types of electromagnetic fields. Moreover, the proposed mathematical description can be used to compare the influence of different types of magnetic fields on heat transfer operations.

Keywords: heat transfer, magnetically assisted reactor, dynamical analysis, transient function

Procedia PDF Downloads 146
370 Removal of Heavy Metals Pb, Zn and Cu from Sludge Waste of Paper Industries Using Biosurfactant

Authors: Nurul Hidayati

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Increasing public awareness of environmental pollution influences the search and development of technologies that help in clean up of organic and inorganic contaminants such as metals. Sludge waste of paper industries as toxic and hazardous material from specific source contains Pb, Zn, and Cu metal from waste soluble ink. An alternative and eco-friendly method of remediation technology is the use of biosurfactants and biosurfactant-producing microorganisms. Soil washing is among the methods available to remove heavy metal from sediments. The purpose of this research is to study effectiveness of biosurfactant with concentration = CMC for the removal of heavy metals, lead, zinc and copper in batch washing test under four different biosurfactant production by microbial origin. Pseudomonas putida T1(8), Bacillus subtilis 3K, Acinetobacter sp, and Actinobacillus sp was grown on mineral salt medium that had been already added with 2% concentration of molasses that it is a low cost application. The samples were kept in a shaker 120 rpm at room temperature for 3 days. Supernatants and sediments of sludge were separated by using a centrifuge and samples from supernatants were measured by atomic absorption spectrophotometer. The highest removal of Pb was up to 14,04% by Acinetobacter sp. Biosurfactant of Pseudomonas putida T1(8) have the highest removal for Zn and Cu up to 6,5% and 2,01% respectively. Biosurfactants have a role for removal process of the metals, including wetting, contact of biosurfactant to the surface of the sediments and detachment of the metals from the sediment. Biosurfactant has proven its ability as a washing agent in heavy metals removal from sediments, but more research is needed to optimize the process of removal heavy metals.

Keywords: biosurfactant, removal of heavy metals, sludge waste, paper industries

Procedia PDF Downloads 293
369 Silver Nanoparticles Loaded Cellulose Nanofibers (Cnf)/mesoporous Bioactive Glass Hydrogels For Periodontitis Treatment

Authors: Anika Pallapothu

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Periodontitis, a severe gum disease, poses a significant threat to the integrity of bone and soft tissues supporting teeth, primarily initiated by bacterial accumulation around the gum line. Conventional treatments like scaling/root planning and plaque removal are widely employed, but integrating modern technologies such as nanotechnology holds promise for innovative therapeutic approaches. This study explores the utilization of silver nanoparticles encapsulated within cellulose nanofiber (CNF) and mesoporous bioactive glass hydrogel matrices for periodontitis management. Silver nanoparticles exhibit potent antimicrobial properties by disrupting microbial cell membranes, inducing reactive oxygen species (ROS) generation, and interfering with vital cellular processes like ATP production and nucleic acid synthesis. Mesoporous bioactive glass, renowned for its high surface area, osteoconductive, and bioactivity, presents a favorable platform for pharmaceutical applications. Incorporating CNF enhances the properties of the hydrogel due to its biocompatibility, biodegradability, and water absorption capacity. The proposed composite material is anticipated to exert beneficial effects in periodontitis treatment by demonstrating antibacterial and anti-inflammatory activities, offering a promising avenue for future therapeutic interventions.

Keywords: periodontitis, cellulose nanofibers, silver nanoparticles, mesoporous bioactive glass, antibacterial activity, anti-inflammatory activity

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368 Assessment of Escherichia coli along Nakibiso Stream in Mbale Municipality, Uganda

Authors: Abdul Walusansa

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The aim of this study was to assess the level of microbial pollution along Nakibiso stream. The study was carried out in polluted waters of Nakibiso stream, originating from Mbale municipality and running through ADRA Estates to Namatala Wetlands in Eastern Uganda. Four sites along the stream were selected basing on the activities of their vicinity. A total of 120 samples were collected in sterile bottles from the four sampling locations of the stream during the wet and dry seasons of the year 2011. The samples were taken to the National water and Sewerage Cooperation Laboratory for Analysis. Membrane filter technique was used to test for Erischerichia coli. Nitrogen, Phosphorus, pH, dissolved oxygen, electrical conductivity, total suspended solids, turbidity and temperature were also measured. Results for Nitrogen and Phosphorus for sites; 1, 2, 3 and 4 were 1.8, 8.8, 7.7 and 13.8 NH4-N mg/L; and 1.8, 2.1, 1.8 and 2.3 PO4-P mg/L respectively. Basing on these results, it was estimated that farmers use 115 and 24 Kg/acre of Nitrogen and Phosphorus respectively per month. Taking results for Nitrogen, the same amount of Nutrients in artificial fertilizers would cost $ 88. This shows that reuse of wastewater has a potential in terms of nutrients. The results for E. coli for sites 1, 2, 3 and 4 were 1.1 X 107, 9.1 X 105, 7.4 X 105, and 3.4 X 105 respectively. E. coli hence decreased downstream with statistically significant variations between sites 1 and 4. Site 1 had the highest mean E.coli counts. The bacterial contamination was significantly higher during the dry season when more water was needed for irrigation. Although the water had the potential for reuse in farming, bacterial contamination during both seasons was higher than 103 FC/100ml recommended by WHO for unrestricted Agriculture.

Keywords: E. coli, nitrogen, phosphorus, water reuse, waste water

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367 Bacterial Cellulose/Silver-Doped Hydroxyapatite Composites for Tissue Engineering Application

Authors: Adrian Ionut Nicoara, Denisa Ionela Ene, Alina Maria Holban, Cristina Busuioc

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At present, the development of materials with biomedical applications is a domain of interest that will produce a full series of benefits in engineering and medicine. In this sense, it is required to use a natural material, and this paper is focused on the development of a composite material based on bacterial cellulose – hydroxyapatite and silver nanoparticles with applications in hard tissue. Bacterial cellulose own features like biocompatibility, non-toxicity character and flexibility. Moreover, the bacterial cellulose can be conjugated with different forms of active silver to possess antimicrobial activity. Hydroxyapatite is well known that can mimic at a significant level the activity of the initial bone. The material was synthesized by using an ultrasound probe and finally characterized by several methods. Thereby, the morphological properties were analyzed by using Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). Because the synthesized material has medical application in restore the tissue and to fight against microbial invasion, the samples were tested from the biological point of view by evaluating the biodegradability in phosphate-buffered saline (PBS) and simulated body fluid (SBF) and moreover the antimicrobial effect was performed on Gram-positive bacterium Staphylococcus aureus, Gram-negative bacterium Escherichia coli, and fungi Candida albicans. The results reveal that the obtained material has specific characteristics for bone regeneration.

Keywords: bacterial cellulose, biomaterials, hydroxyapatite, scaffolds materials

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366 Understanding Integrated Removal of Heavy Metals, Organic Matter and Nitrogen in a Constructed Wetland System Receiving Simulated Landfill Leachate

Authors: A. Mohammed, A. Babatunde

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This study investigated the integrated removal of heavy metals, organic matter and nitrogen from landfill leachate using a novel laboratory scale constructed wetland system. The main objectives of this study were: (i) to assess the overall effectiveness of the constructed wetland system for treating landfill leachate; (ii) to examine the interactions and impact of key leachate constituents (heavy metals, organic matter and nitrogen) on the overall removal dynamics and efficiency. The constructed wetland system consisted of four stages operated in tidal flow and anoxic conditions. Results obtained from 215 days of operation have demonstrated extraordinary heavy metals removal up to 100%. Analysis of the physico- chemical data reveal that the controlling factors for metals removal were the anoxic condition and the use of the novel media (dewatered ferric sludge which is a by-product of drinking water treatment process) as the main substrate in the constructed wetland system. Results show that the use of the ferric sludge enhanced heavy metals removal and brought more flexibility to simultaneous nitrification and denitrification which occurs within the microbial flocs. Furthermore, COD and NH4-N were effectively removed in the system and this coincided with enhanced aeration in the 2nd and 3rd stages of the constructed wetland system. Overall, the results demonstrated that the ferric dewatered sludge constructed wetland system would be an effective solution for integrated removal of pollutants from landfill leachates.

Keywords: constructed wetland, ferric dewatered sludge, heavy metals, landfill leachate

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365 Extracellular Phytase from Lactobacillus fermentum spp KA1: Optimization of Enzyme Production and Its Application for Improving the Nutritional Quality of Rice Bran

Authors: Neha Sharma, Kanthi K. Kondepudi, Naveen Gupta

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Phytases are phytate specific phosphatases catalyzing the step-wise dephosphorylation of phytate, which acts as an anti-nutritional factor in food due to its strong binding capacity to minerals. In recent years microbial phytases have been explored for improving nutritional quality of food. But the major limitation is acceptability of phytases from these microorganisms. Therefore, efforts are being made to isolate organisms which are generally regarded as safe for human consumption such as Lactic Acid Bacteria (LAB). Phytases from these organisms will have an edge over other phytase sources due to its probiotic attributes. Only few LAB have been reported to give phytase activity that too is generally seen as intracellular. LAB producing extracellular phytase will be more useful as it can degrade phytate more effectively. Moreover, enzyme from such isolate will have application in food processing also. Only few species of Lactobacillus producing extracellular phytase have been reported so far. This study reports the isolation of a probiotic strain of Lactobacillus fermentum spp KA1 which produces extracellular phytase. Conditions for the optimal production of phytase have been optimized and the enzyme production resulted in an approximately 13-fold increase in yield. The phytate degradation potential of extracellular phytase in rice bran has been explored and conditions for optimal degradation were optimized. Under optimal conditions, there was 43.26% release of inorganic phosphate and 6.45% decrease of phytate content.

Keywords: Lactobacillus, phytase, phytate reduction, rice bran

Procedia PDF Downloads 162
364 Assessment of Knowledge, Attitudes and Practices of Street Vendors in Mangaung Metro South Africa

Authors: Gaofetoge Lenetha, Malerato Moloi, Ntsoaki Malebo

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Microbial contamination of ready-to-eat foods and beverages sold by street vendors has become an important public health issue. In developing countries including South Africa, health risks related to such kinds of foods are thought to be common. Thus, this study assessed knowledge, attitude and practices of street food vendors. Street vendors in the city of Mangaung Metro were investigated in order to assess their knowledge, attitudes and handling practices. A semi-structured questionnaire and checklist were used in interviews to determine the status of the vending sites and associa. ted food-handling practices. Data was collected by means of a face-to-face interview. The majority of respondents were black females. Hundred percent (100%) of the participants did not have any food safety training. However, street vendors showed a positive attitude towards food safety. Despite the positive attitude, vendors showed some non-compliance when it comes to handling food. During the survey, it was also observed that the vending stalls lack basic infrastructures like toilets and potable water that is currently a major problem. This study indicates a need for improvements in the environmental conditions at these sites to prevent foodborne diseases. Moreover, based on the results observed food safety and food hygiene training or workshops for street vendors are highly recommended.

Keywords: food hygiene, foodborne illnesses, food safety, Street foods

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363 A Combined Activated Sludge-Filtration-Ozonation Process for Abattoir Wastewater Treatment

Authors: Pello Alfonso-Muniozguren, Madeleine Bussemaker, Ralph Chadeesingh, Caryn Jones, David Oakley, Judy Lee, Devendra Saroj

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Current industrialized livestock agriculture is growing every year leading to an increase in the generation of wastewater that varies considerably in terms of organic content and microbial population. Therefore, suitable wastewater treatment methods are required to ensure the wastewater quality meet regulations before discharge. In the present study, a combined lab scale activated sludge-filtration-ozonation system was used to treat a pre-treated abattoir wastewater. A hydraulic retention time of 24 hours and a solid retention time of 13 days were used for the activated sludge process, followed by a filtration step (4-7 µm) and using ozone as tertiary treatment. An average reduction of 93% and 98% was achieved for Chemical Oxygen Demand (COD) and Biological Oxygen Demand (BOD), respectively, obtaining final values of 128 mg/L COD and 12 mg/L BOD. For the Total Suspended Solids (TSS), the average reduction increased to 99% in the same system, reducing the final value down to 3 mg/L. Additionally, 98% reduction in Phosphorus (P) and a complete inactivation of Total Coliforms (TC) was obtained after 17 min ozonation time. For Total Viable Counts (TVC), a drastic reduction was observed with 30 min ozonation time (6 log inactivation) at an ozone dose of 71 mg O3/L. Overall, the combined process was sufficient to meet discharge requirements without further treatment for the measured parameters (COD, BOD, TSS, P, TC, and TVC).

Keywords: abattoir waste water, activated sludge, ozone, waste water treatment

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362 Biodegradation of Triclosan and Tetracycline in Sewage Sludge by Pleurotus Ostreatus Fungal Pellets

Authors: Ayda Maadani Mallak, Amir lakzian, Elham Khodaverdi, Gholam Hossein Haghnia

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The use of pharmaceuticals and personal care products such as antibiotics and antibacterials has been increased in recent years. Since the major part of consumed compounds remains unchanged in the wastewater treatment plant, they will easily find their way into the human food chain following the land use of sewage sludge (SS). Biological treatment of SS is one the most effective methods for expunging contaminants. White rot fungi, due to their ligninolytic enzymes, are extensively used to degrade organic compounds. Among all three different morphological forms and growth patterns of filamentous fungi (mycelia, clumps, and pellets), fungal pellet formation has been the subject of interest in industrial bioprocesses. Therefore this study was aimed to investigate the uptake of tetracycline (TC) and triclosan (TCS) by radish plant (Raphanus sativus) from soil amended with untreated and pretreated SS by P. ostreatus fungal pellets under greenhouse conditions. The experimental soil was amended with 1) Contaminated SS with TC at a concentration of 100 mgkg-1 and pretreated by fungal pellets, 2) Contaminated SS with TC at 100 mgkg-1 and untreated with fungal pellets, 3) Contaminated SS with TCS at a concentration of 50 mgkg-1 and pretreated by fungal pellets, 4) contaminated SS with TCS at 50 mgkg-1 and untreated with fungal pellets. An uncontaminated and untreated SS-amended soil also was considered as control treatment. An AB SCIEX 3200 QTRAP LC-MS/MS system was used in order to analyze the concentration of TC and TCS in plant tissues and soil medium. Results of this study revealed that the presence of TC and TCS in SS-amended soil decreased the radish biomass significantly. The reduction effect of TCS on dry biomass of shoot and root was 39 and 45% compared to controls, whereas for TC, the reduction percentage for shoot and root was 27 and 40.6%, respectively. However, fungal treatment of SS by P. ostreatus pellets reduced the negative effect of both compounds on plant biomass remarkably, as no significant difference was observed compared to control treatments. Pretreatment of SS with P. ostreatus also caused a significant reduction in translocation factor (concentration in shoot/root), especially for TC compound up to 32.3%, whereas this reduction for TCS was less (8%) compared to untreated SS. Generally, the results of this study confirmed the positive effect of using fungal pellets in SS amendment to decrease TC and TCS uptake by radish plants. In conclusion, P. ostreatus fungal pellets might provide future insights into bioaugmentation to remove antibiotics from environmental matrices.

Keywords: antibiotic, fungal pellet, sewage sludge, white-rot fungi

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361 In Vitro Antibacterial Effect of Hydroalcoholic Extract of Lawsonia Inermis, Malva Sylvestris and Boswellia Serrata on Aggregatibacter Actinomycetemcomitans

Authors: Surena V.

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Background and Aim: Periodontal diseases are among the most common infectious diseases all around the world, even in developed countries. Considering the increased rate of microbial resistance to antibiotics and the chemical side effects of antibiotics and antiseptics used for the treatment of periodontal disease, there is a need for an alternative antimicrobial agent with fewer complications. Medicinal herbs have recently become popular as antimicrobial and preventive agents. This study aimed to assess the antibacterial effects of hydroalcoholic extracts of Lawsonia inermis, Malva sylvestris and Boswellia serrata on Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans). Materials and Methods: Hydroalcoholic extracts of the three medicinal plants were obtained by the maceration technique and A. actinomycetemcomitans was cultured. The antimicrobial efficacy of the three medicinal plants was compared with that of 0.2% chlorhexidine (CHX) according to the CLSI protocol using agar disc diffusion and broth microdilution techniques. All tests were repeated three times. Results: Hydroalcoholic extracts of all three plants had antimicrobial activity against A. actinomycetemcomitans. The minimum inhibitory concentration (MIC) of Lawsonia inermis, Malva sylvestris, and Boswellia serrata was 78.1, 156.2, and 1666 µg/mL with no significant difference between them. The MIC of CHX was 3.33 µg/mL, which was significantly higher than that of Boswellia serrata extract. Conclusion: Given that, further in vivo studies confirm other properties of these extracts and their safety in terms of cytotoxicity and mutagenicity, hydroalcoholic extracts of Lawsonia inermis and Malva sylvestris may be used in mouthwashes or local delivery systems to affect periodontal biofilm.

Keywords: actinobacilus actinomycetem commitans, lawsonia inermis, malva sylvestris, boswellia serrata

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360 Preservation of Traditional Algerian Sausage Against Microbial Activity by the Garlic (Allium Sativum L.)

Authors: Abed Hannane, Rouag Noureddine

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The present study aims to evaluate the association of fresh garlic (Allium sativum L.) and storage at 4°C in preserving the microbiological, nutritional, and sanitary quality of Merguez-type sausages prepared and sold locally from meat offal. The analysis focused on the evaluation of the microbiological quality of fifteen samples randomly taken from several butcheries in the wilaya of BBA, eastern Algeria. The bacteriological analysis revealed the presence of 6.88.10⁵ CFU/g of total aerobic bacteria, 5.39.10⁵ CFU/g of total coliforms, 2.23.10⁵ CFU/g of fecal coliforms, 2.43.103 CFU/g of Escherichia coli and 1.8.10⁵ CFU/g of coagulase-positive staphylococci, values higher than Algerian standards. The addition of fresh garlic as an antibacterial preservative at concentrations of 0.06, 0.12, 0.18, and 0.24 g/g to ground beef samples and stored in the refrigerator at 4°C for 15 days. The addition of garlic to Merguez made it possible to significantly reduce the presence of different bacterial groups during their refrigerated storage, compared to untreated meat, bringing it below the standards defined in the matter. Thus, the use of garlic as a food additive at a concentration of 0.12 g/g was sufficient to obtain levels according to Algerian standards equal to 1.8.10⁴ CFU/g of total aerobic bacteria, 9.48.10³ CFU/ g of total coliforms, 3.68.10³ UFC/g fecal coliforms, 4.56.10² UFC/g of E.coli 2.39.10⁴ UFC/g of coagulase-positive staphylococci. It is clear that thanks to the addition of garlic to Merguez, the sanitary quality has been improved by reducing the aerobic bacterial load and increasing the shelf life at 4°C.

Keywords: antimicrobial effect, garlic, sausage, storage

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359 Isolation of a Bacterial Community with High Removal Efficiencies of the Insecticide Bendiocarb

Authors: Eusebio A. Jiménez-Arévalo, Deifilia Ahuatzi-Chacón, Juvencio Galíndez-Mayer, Cleotilde Juárez-Ramírez, Nora Ruiz-Ordaz

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Bendiocarb is a known toxic xenobiotic that presents acute and chronic risks for freshwater invertebrates and estuarine and marine biota; thus, the treatment of water contaminated with the insecticide is of concern. In this paper, a bacterial community with the capacity to grow in bendiocarb as its sole carbon and nitrogen source was isolated by enrichment techniques in batch culture, from samples of a composting plant located in the northeast of Mexico City. Eight cultivable bacteria were isolated from the microbial community, by PCR amplification of 16 rDNA; Pseudoxanthomonas spadix (NC_016147.2, 98%), Ochrobacterium anthropi (NC_009668.1, 97%), Staphylococcus capitis (NZ_CP007601.1, 99%), Bosea thiooxidans. (NZ_LMAR01000067.1, 99%), Pseudomonas denitrificans. (NC_020829.1, 99%), Agromyces sp. (NZ_LMKQ01000001.1, 98%), Bacillus thuringiensis. (NC_022873.1, 97%), Pseudomonas alkylphenolia (NZ_CP009048.1, 98%). NCBI accession numbers and percentage of similarity are indicated in parentheses. These bacteria were regarded as the isolated species for having the best similarity matches. The ability to degrade bendiocarb by the immobilized bacterial community in a packed bed biofilm reactor, using as support volcanic stone fragments (tezontle), was evaluated. The reactor system was operated in batch using mineral salts medium and 30 mg/L of bendiocarb as carbon and nitrogen source. With this system, an overall removal efficiency (ηbend) rounding 90%, was reached.

Keywords: bendiocarb, biodegradation, biofilm reactor, carbamate insecticide

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358 Kinetic Study of C₃N₄/CuWO₄: Photocatalyst towards Solar Light Inactivation of Mixed Populated Bacteria

Authors: Rimzhim Gupta, Bhanupriya Boruah, Jayant M. Modak, Giridhar Madras

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Microbial contamination is one of the major concerns in the field of water treatment. AOP (advanced oxidation processes) is well-established method to resolve the issue of removal of contaminants in water. A Z-scheme composite g-C₃N₄/CuWO₄ was synthesized by sol-gel method for the photocatalytic inactivation of a mixed population of Gram-positive bacteria (S. aureus) and Gram-negative bacteria (E. coli). The photoinactivation was observed for different types of bacteria in the same medium together and individually in the absence of the nutrients. The lattice structures and phase purities were determined by X-ray diffraction. For morphological and topographical features, scanning electron microscopy and transmission electron microscopy analyses were carried out. The band edges of the semiconductor (valence band and conduction band) were determined by ultraviolet photoelectron microscopy. The lifetime of the charge carriers and band gap of the semiconductors were determined by time resolved florescence spectroscopy and diffused reflectance spectroscopy, respectively. The effect of weight ratio of C₃N₄ and CuWO₄ was observed by performing photocatalytic experiments. To investigate the exact mechanism and major responsible radicals for photocatalysis, scavenger studies were performed. The rate constants and order of the inactivation reactions were obtained by power law kinetics. For E. coli and S. aureus, the order of reaction and rate constants are 1.15, 0.9 and 1.39 ± 0.03 (CFU/mL)⁻⁰.¹⁵ h⁻¹, 47.95 ± 1.2 (CFU/mL)⁰.¹ h⁻¹, respectively.

Keywords: z-scheme, E. coli, S. aureus, sol-gel

Procedia PDF Downloads 126
357 Effects of Fermentation Techniques on the Quality of Cocoa Beans

Authors: Monday O. Ale, Adebukola A. Akintade, Olasunbo O. Orungbemi

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Fermentation as an important operation in the processing of cocoa beans is now affected by the recent climate change across the globe. The major requirement for effective fermentation is the ability of the material used to retain sufficient heat for the required microbial activities. Apart from the effects of climate on the rate of heat retention, the materials used for fermentation plays an important role. Most Farmers still restrict fermentation activities to the use of traditional methods. Improving on cocoa fermentation in this era of climate change makes it necessary to work on other materials that can be suitable for cocoa fermentation. Therefore, the objective of this study was to determine the effects of fermentation techniques on the quality of cocoa beans. The materials used in this fermentation research were heap-leaves (traditional), stainless steel, plastic tin, plastic basket and wooden box. The period of fermentation varies from zero days to 10 days. Physical and chemical tests were carried out for variables in quality determination in the samples. The weight per bean varied from 1.0-1.2 g after drying across the samples and the major color of the dry beans observed was brown except with the samples from stainless steel. The moisture content varied from 5.5-7%. The mineral content and the heavy metals decreased with increase in the fermentation period. A wooden box can conclusively be used as an alternative to heap-leaves as there was no significant difference in the physical features of the samples fermented with the two methods. The use of a wooden box as an alternative for cocoa fermentation is therefore recommended for cocoa farmers.

Keywords: fermentation, effects, fermentation materials, period, quality

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356 Bioprospecting of Marine Actinobacteria: The Leading Way for Industrially Important Enzymes and Bioactive Natural Products

Authors: Ramesh Subramani, Mathivanan Narayanasamy, William Aalbersberg

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It is well accepted by last 35 years of research and on-going programmes that marine environment harbours abundant and unique biodiversity, which is currently playing as an important source in bioprospecting. It has become apparent that marine microorganisms are lead in the biodiscovery. Among marine organisms, actinobacteria are a target phylum for discovering novel antibiotics against increasing the multi-drug resistant human pathogens because of these taxa representing for novel genera and species. Marine actinomycetes are a proven source of new antibiotic leads and novel enzymes with important industrial applications. A total of 183 streptomycete and 25 non-streptomycete strains were isolated from different marine samples collected from north-eastern part of the Indian Ocean. Among them, 111 isolates displayed antibacterial activity against human pathogens and 151 exhibited antifungal activity against phytopathogens. Importantly, most of them produced various extracellular enzymes and 58 of them produced exopolysaccharides. Totally eight small bioactive compounds and a thermostable alkaline protease have been purified from a selected strain, Streptomyces fungicidicus. Besides, our on-going studies on non-streptomycete strains (rare actinomycetes) are most likely promising resource for new and unique compounds against current emerging drug-resistant pathogens. We have just recognised the chemical diversity in marine microorganisms. Therefore it is worthwhile to continue the exploration of marine microorganisms for new drug leads, novel enzymes and other bioprospecting research.

Keywords: bioactive compounds, industrial enzymes, marine actinobacteria, microbial metabolites, marine natural products

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355 Management of Fungal Diseases of Onion (Allium cepa L.) by Using Plant Extracts

Authors: Shobha U. Jadhav, R. S. Saler

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Onion is most Important Vegetable crop grown throughout the world. Onion suffers from pest and fungal diseases but the fungicides cause pollution and disturb microbial balance of soil. Under integrated fungal disease management programme cost effective and eco- friendly component like plant extract are used to control plant pathogens. Alternaria porri, Fusarium oxysporium, Stemphylium vesicarium are soil borne pathogens of onion. Effect of three different plant extract (Datura metel, Pongamia pinnata, Ipomoea palmata) at five different concentration Viz, 10,25,50,75 and 100 percentage on these pathogens was studied by food poisoning techniquie. Detura metal gave 94.73% growth of Alternaria porri at 10% extract concentraton and 26.31% growth in 100% extract concentration. As compared to Fusarium oxysporium, and Stemphylium vesicarium, Alternaria porri give good inhibitory response. In Pongamia pinnata L. at 10% extract concentration 84.21% growth and at 100% extract concentration 36.84% growth of Stemphylium vesicarium was observed. Stemphylium vesicarium give good in inhibitory response as compared to Alternaria porri and Fusarium oxysporium. Ipomoea palmata in 10% extract concentration 92% growth and in 100% extract concentration 40% growth of Fusarium oxysporium was recorded. Fusarium oxysporium give good inhibitory response as compared to Alternaria porri and, Stemphylium vesicarium.

Keywords: pathogen, onion, plant extract, Allium cepa L.

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354 Bioremediation Potential in Recalcitrant Areas of PCE in Alluvial Fan Deposits

Authors: J. Herrero, D. Puigserver, I. Nijenhuis, K. Kuntze, J. M. Carmona

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In the transition zone between aquifers and basal aquitards, the perchloroethene (PCE)-pools are more recalcitrant than those elsewhere in the aquifer. Although biodegradation of chloroethenes occur in this zone, it is a slow process and a remediation strategy is needed. The aim of this study is to demonstrate that combined strategy of biostimulation and in situ chemical reduction (ISCR) is more efficient than the two separated strategies. Four different microcosm experiments with sediment and groundwater of a selected field site where an aged pool exists at the bottom of a transition zone were designed under i) natural conditions, ii) biostimulation with lactic acid, iii) ISCR with zero-value iron (ZVI) and under iv) a combined strategy with lactic acid and ZVI. Biotic and abiotic dehalogenation, terminal electron acceptor processes and evolution of microbial communities were determined for each experiment. The main results were: i) reductive dehalogenation of PCE-pools occurs under sulfate-reducing conditions; ii) biostimulation with lactic acid supports more pronounced reductive dehalogenation of PCE and trichloroethene (TCE), but results in an accumulation of 1,2-cis-dichloroethene (cDCE); iii) ISCR with ZVI produces a sustained dehalogenation of PCE and its metabolites iv) combined strategy of biostimulation and ISCR results in a fast dehalogenation of PCE and TCE and a sustained dehalogenation of cisDCE. These findings suggest that biostimulation and ISCR with ZVI are the most suitable strategies for a complete reductive dehalogenation of PCE-pools in the transition zone and further to enable the dissolution of dense non-aqueous phase liquids.

Keywords: aged PCE-pool, anaerobic microcosm experiment, biostimulation, in situ chemical reduction, natural attenuation

Procedia PDF Downloads 169
353 New Applications of Essential Oils: Edible Packaging Material for Food Supplements

Authors: Roxana Gheorghita, Gheorghe Gutt

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Environmental pollution due to non-degradation of packaging from the food and pharmaceutical industry is reaching increasingly alarming levels. The packaging used for food supplements is usually composed of successive layers of synthetic materials, conventional, glue, and paint. The situation is becoming more and more problematic as the population, according to statistics, uses food supplements more and more often. The solution can be represented by edible packaging, completely biodegradable, and compostable. The tested materials were obtained from biopolymers, agar, carrageenan, and alginate, in well-established quantities and plasticized with glycerol. Rosemary, thyme, and oregano essential oils have been added in varying proportions. The obtained films are completely water-soluble in hot liquids (with a temperature of about 80° C) and can be consumed with the product contained. The films were glossy, pleasant to the touch, thin (thicknesses between 32.8 and 52.8 μm), transparent, and with a pleasant smell, specific to the added essential oil. Tested for microbial evaluation, none of the films indicated the presence of E. coli, S. aureus, enterobacteria, coliform bacteria, yeasts, or molds. This aspect can also be helped by the low values of the water activity index (located between 0.546 and 0.576). The mechanical properties indicated that the material became more resistant with the addition of essential oil, the best values being recorded by the addition of oregano. The results obtained indicate the possibility of using biopolymer-based films with the addition of rosemary, thyme, and oregano essential oil, for wrapping food supplements, thus replacing conventional packaging, multilayer, impossible to sort and recycle.

Keywords: edible films, food supplements, oregano, rosemary, thyme

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352 Production of Biotechnological Chondroitin from Recombinant E, Coli K4 Strains on Renewable Substrates

Authors: Donatella Cimini, Sergio D’ambrosio, Saba Sadiq, Chiara Schiraldi

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Chondroitin sulfate (CS), as well as modified CS, and unsulfated chondroitin, are largely applied in research today. CS is a linear glycosaminoglycan normally present in cartilage-rich tissues and bones in the form of proteoglycans decorated with sulfate groups in different positions. CS is used as an effective non-pharmacological alternative for the treatment of osteoarthritis, and other potential applications in the biomedical field are being investigated. Some bacteria, such as E. coli K4, produce a polysaccharide that is a precursor of CS (unsulfated chondroitin). This work focused on the construction of integrative E. coli K4 recombinant strains overexpressing genes (kfoA, kfoF, pgm and galU in different combinations) involved in the biosynthesis of the nucleotide sugars necessary for polysaccharide synthesis. Strain growth and polymer production were evaluated using renewable waste materials as substrates in shake flasks and small-scale batch fermentation processes. Results demonstrated the potential to replace pure sugars with cheaper medium components to establish environmentally sustainable and cost-effective production routes for potential industrial development. In fact, although excellent fermentation results have been described so far by employing strains that naturally produce chondroitin-like polysaccharides on semi-defined media, there is still the need to reduce manufacturing costs by providing a cost-effective biotechnological alternative to currently used animal-based extraction procedures.

Keywords: E. coli K4, chondroitin, microbial cell factories, glycosaminoglycans, renewable resources

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351 Exploring the Intricate Microbiology of Street Cuisine: Delving into Potential Dangers in Order to Enhance Safety and Quality

Authors: Raana Babadi Fathipour

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Street foods hold a significant place in the tapestry of socioeconomic and cultural norms, beloved across the globe. Serving as a convenient and affordable option for city dwellers seeking nourishment, these culinary delights also serve as a vital source of income for vendors, particularly women. Additionally, street food acts as a mirror reflecting traditional local customs and practices, an element that draws tourists to experience the authenticity of a culture firsthand. Despite its many virtues, concerns have emerged regarding the microbiological safety of street food worldwide. Often prepared and sold in subpar conditions without proper oversight or regulation, street food has become synonymous with potential health risks. The presence of elevated levels of fecal indicator bacteria and various pathogens in these unregulated delicacies further perpetuates anxieties surrounding their consumption. This analysis delves into the intricate microbiological intricacies inherent in street food, shedding light on the pertinent safety concerns and prevalent pathogens. Additionally, it elaborates on the worldwide standing of this vital economic endeavor. Moreover, it advocates for the adoption of molecular detection techniques over conventional culture-based methods to gain a more comprehensive grasp of the true microbial risks posed by street cuisine. Acknowledgment marks the initial step towards resolving any given issue.

Keywords: foodborne pathogens, microbiological safety, street food, viruses

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350 Isolation, Identification and Characterization of the Bacteria and Yeast from the Fermented Stevia Extract

Authors: Asato Takaishi, Masashi Nasuhara, Ayuko Itsuki, Kenichi Suga

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Stevia (Stevia rebaudiana Bertoni) is a composite plant native to Paraguay. Stevia sweetener is derived from a hot water extract of Stevia (Stevia extract), which has some effects such as histamine decomposition, antioxidative effect, and blood sugar level-lowering function. The steviol glycosides in the Stevia extract are considered to contribute to these effects. In addition, these effects increase by the fermentation. However, it takes a long time for fermentation of Stevia extract and the fermentation liquid sometimes decays during the fermentation process because natural fermentation method is used. The aim of this study is to perform the fermentation of Stevia extract in a shorter period, and to produce the fermentation liquid in stable quality. From the natural fermentation liquid of Stevia extract, the four strains of useful (good taste) microorganisms were isolated using dilution plate count method and some properties were determined. The base sequences of 16S rDNA and 28S rDNA revealed three bacteria (two Lactobacillus sp. and Microbacterium sp.) and one yeast (Issatchenkia sp.). This result has corresponded that several kinds of lactic bacterium such as Lactobacillus pentosus and Lactobacillus buchneri were isolated from Stevia leaves. Liquid chromatography/mass spectrometory (LC/MS/MS) and High-Performance Liquid Chromatography (HPLC) were used to determine the contents of steviol glycosides and neutral sugars. When these strains were cultured in the sterile Stevia extract, the steviol and stevioside were increased in the fermented Stevia extract. So, it was suggested that the rebaudioside A and the mixture of steviol glycosides in the Stevia extract were decomposed into stevioside and steviol by microbial metabolism.

Keywords: fermentation, lactobacillus, Stevia, steviol glycosides, yeast

Procedia PDF Downloads 516
349 Synthesis of Plant-Mediated Silver Nanoparticles Using Erythrina indica Extract and Evaluation of Their Anti-Microbial Activities

Authors: Chandra Sekhar Singh, P. Chakrapani, B. Arun Jyothi, A. Roja Rani

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The green synthesis of metallic nanoparticles (NPs) involves biocompatible ingredients under physiological conditions of temperature and pressure. Moreover, the biologically active molecules involved in the green synthesis of NPs act as functionalizing ligands, making these NPs more suitable for biomedical applications. Among the most important bioreductants are plant extracts, which are relatively easy to handle, readily available, low cost, and have been well explored for the green synthesis of other nanomaterials. Various types of metallic NPs have already been synthesized using plant extracts. They have wide applicability in various areas such as electronics, catalysis, chemistry, energy, and medicine. Metallic nanoparticles are traditionally synthesized by wet chemical techniques, where the chemicals used are quite often toxic and flammable. In our study, we were described a cost effective and environment friendly technique for green synthesis of silver nanoparticles from 1mM AgNO3 solution through the aqueous extract of Erythrina indica as reducing as well as capping agent. Nanoparticles were characterized using UV–Vis absorption spectroscopy, FTIR, XRD, X-ray diffraction, SEM and TEM analysis showed the average particle size of 30 nm as well as revealed their spherical structure. Further these biologically synthesized nanoparticles were found to be highly toxic against different human pathogens viz. two Gram positive namely Klebsiella pneumonia and Bacillus subtilis bacteria and two were Gram negative bacteria namely Staphylococcus aureus and Escherichia coli (E. coli). This is for the first time reporting that Erythrina indica plant extract was used for the synthesis of nanoparticles.

Keywords: silver nanoparticles, green synthesis, antibacterial activity, FTIR, TEM, SEM

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348 CO2 Capture in Porous Silica Assisted by Lithium

Authors: Lucero Gonzalez, Salvador Alfaro

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Carbon dioxide (CO2) and methane (CH4) are considered as the compounds with higher abundance among the greenhouse gases (CO2, NOx, SOx, CxHx, etc.), due to its higher concentration, this two gases have a greater impact in the environment pollution and provokes global warming. So, recovery, disposal and subsequent reuse, are of great interest, especially from the ecological and health perspective. By one hand, porous inorganic materials are good candidates to capture gases, because these type of materials are higher stability from the point view of thermal, chemical and mechanical under adsorption gas processes. By another hand, during the design and the synthetic preparation of the porous materials is possible add other intrinsic properties (physicochemical and structural) by adding chemical compounds as dopants or using structured directed agents or surfactants to improve the porous structure, the above features allow to have alternative materials for separation, capture and storage of greenhouse gases. In this work, ordered mesoporous materials base silica were prepared using Surfynol as surfactant. The surfactant micelles are commonly used as self-assembly templates for the development of new structure porous silica’s, adding a variety of textures and structures. By another hand, the Surfynol is a commercial surfactant, is non-ionic, for that is necessary determine its critical micelles concentration (cmc) by the pyrene I1/I3 ratio method, before to prepare silica particles. One time known the CMC, a precursor gel was prepared via sol-gel process at room temperature using TEOS as silica precursor, NH4OH as catalyst, Surfynol as template and H2O as solvent. Then, the gel precursor was treatment hydrothermally in a Teflon-lined stainless steel autoclave with a volume of 100 mL and kept at 100 ºC for 24 h under static conditions in a convection oven. After that, the porous silica particles obtained were impregnated with lithium to improve the CO2 adsorption capacity. Then the silica particles were characterized physicochemical, morphology and structurally, by XRD, FTIR, BET and SEM techniques. The thermal stability and the CO2 adsorption capacity was evaluated by thermogravimetric analysis (TGA). According the results, we found that the Surfynol is a good candidate to prepare silica particles with an ordered structure. Also the TGA analysis shown that the particles has a good thermal stability in the range of 250 °C and 800 °C. The best materials had, the capacity to adsorbing 70 and 90 mg per gram of silica particles and its CO2 adsorption capacity depends on the way to thermal pretreatment of the porous silica before of the adsorption experiments and of the concentration of surfactant used during the synthesis of silica particles. Acknowledgments: This work was supported by SIP-IPN through project SIP-20161862.

Keywords: CO2 adsorption, lithium as dopant, porous silica, surfynol as surfactant, thermogravimetric analysis

Procedia PDF Downloads 241
347 Large Scale Production of Polyhydroxyalkanoates (PHAs) from Waste Water: A Study of Techno-Economics, Energy Use, and Greenhouse Gas Emissions

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

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The biodegradable family of polymers polyhydroxyalkanoates are interesting substitutes 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

Procedia PDF Downloads 419
346 Two Years Retrospective Study of Body Fluid Cultures Obtained from Patients in the Intensive Care Unit of General Hospital of Ioannina

Authors: N. Varsamis, M. Gerasimou, P. Christodoulou, S. Mantzoukis, G. Kolliopoulou, N. Zotos

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Purpose: Body fluids (pleural, peritoneal, synovial, pericardial, cerebrospinal) are an important element in the detection of microorganisms. For this reason, it is important to examine them in the Intensive Care Unit (ICU) patients. Material and Method: Body fluids are transported through sterile containers and enriched as soon as possible with Tryptic Soy Broth (TSB). After one day of incubation, the broth is poured into selective media: Blood, Mac Conkey No. 2, Chocolate, Mueller Hinton, Chapman and Saboureaud agar. The above selective media are incubated directly for 2 days. After this period, if any number of microbial colonies are detected, gram staining is performed. After that, the isolated organisms are identified by biochemical techniques in the automated Microscan system (Siemens) and followed by a sensitivity test on the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by Kirby Bauer-based plate test. Results: In 2017 the Laboratory of Microbiology received 60 samples of body fluids from the ICU. More specifically the Microbiology Department received 6 peritoneal fluid specimens, 18 pleural fluid specimens and 36 cerebrospinal fluid specimens. 36 positive cultures were tested. S. epidermidis was identified in 18 specimens, S. haemolyticus in 6, and E. faecium in 12. Conclusions: The results show low detection of microorganisms in body fluid cultures.

Keywords: body fluids, culture, intensive care unit, microorganisms

Procedia PDF Downloads 178