Search results for: microbial enzymes

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

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Authors: Leelavinothan Pari , Ayyasamy Rathinam

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Diabetes mellitus (DM) is a major and growing public health problem throughout the world. Pterocarpus marsupium (Roxb.) (Family: Fabaceae) is widely used as a traditional medicine to treat various diseases including diabetes. However, the molecular mechanism of Pterocarpus marsupium has not been investigated so far. Two fractions (2.5% and 5%) of extract from the medicinal plant, Pterocarpus marsupium (PME) were conducted in a dose dependent manner in streptozotocin (45 mg/kg b.w.) induced type 2 diabetic rats. Each fraction of PME was administered to diabetic rats intragastrically at a dose of 50, 100 and 200 mg/kg b.w for 45 days. The effective dose 200 mg/kg b.w of 5% fraction was more pronounced in reducing the levels of blood glucose (95.65 mg/dL) and glycosylated hemoglobin (HbA1c) (0.41 mg/g Hb), and increasing the plasma insulin (16.20 µU/mL) level. Moreover, PME (200 mg/kg b.w) significantly ameliorated lipid peroxidation products (thiobarbituric reactive substances, lipid hydroperoxides) enzymatic (superoxide dismutase, catalase and glutathione peroxidase) and non-enzymatic antioxidants (Vitamin C, Vitamin E and reduced glutathione) levels. The altered activities of the key enzymes of lipid metabolism along with the lipid profile in diabetic rats were significantly reverted to near normal levels by the administration of PME 5% 200 mg/kg b.w fraction. PME (200 mg/kg b.w) has the ability to reduce the inflammatory cytokines, such as TNF-α, IL-6 mRNA, as well as protein expression and apoptotic marker, such as caspase-3 enzyme in diabetic hepatic tissue. The above biochemical findings were also supported by histological studies such as improvement in pancreas and liver. Pterocarpus marsupium could effectively reduce the hyperglycemia, oxidative-stress, inflammation and hyperlipedimea in diabetic rats; hence it could be a useful drug in the management of diabetes without any side effects.

Keywords: diabetes mellitus, streptozotocin, Pterocarpus marsupium, lipid peroxidation, Antioxidants, inflammatory cytokines

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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

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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

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Authors: Soufiane Fal, Abderahim Aasfar, Ali Ouhssain, Hasnae Choukri, Abelaziz Smouni, Hicham El Arroussi

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Heavy metals contamination is a major environmental concern around the world. It has a harmful impact on plant productivity and poses a serious risk to humans and animals health. In the present study, the effect of Microalgae Crude Extract (MCE) on tomato growth and nutrients uptake exposed to 2 mM Pb2+ and Cd2+ was investigated. In results, 2 mM Pb2+ and Cd2+ showed a significant reduction of tomatobiomass and perturbation in nutrients absorption. Moreover, MCE application in tomato plant exposed to Pb2+ and Cd2+ showed a significant enhancement of biomass compared to tomato plants under Pb2+ and Cd2+. On the other hand, MCE application favoured heavy metals accumulation in root and inhibited their translocation to shoot as phytostabilisation mechanism. Tomato plants showed biochemical responses to Pb2+ and Cd2+ stress with elevation of scavenging enzymes and molecules such as POD, CAT, SOD, Proline, and polyphenols, etc. In addition, the treatment by MCE showed a significant reduction level of the majority of these parameters. Furthermore, the metabolomic analysis revealed a significant change in important metabolites. Pb2+ and Cd2+ showed decrease in SFA and increase of UFA, VLFA, alkanes, alkenes, sterols, which known accumulated as tolerance and resistance mechanism to heavy metal (H.M) stress. However, MCE treatment showed the inverse of these response to return tomato plants to normal state and enhanced tolerance and resistance to heavy metal stress. In the present study, we emphasized that MCE can alleviate H.M stress, enhance tomato plant growth nutrients absorption and improve biochemical responses.

Keywords: microalgae crude extract, heavy metal stress, nutrient uptake, metabolomic analysis, solanum lycopersicum (Tomato), phytostabilisation

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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

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Authors: Jinyoung Park, Eun Joo Song

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Introduction: Deubiquitinating enzymes (DUBs) are proteases that cleave ubiquitin or ubiquitin-like modifications on substrates. Deubiquitination could regulate cellular physiology, such as signal transduction, DNA damage and repair, and cell cycle progression. Although more than 100 DUBs are encoded in the human and the importance of DUBs has been realized, the functions of most DUBs are unknown. This study aims to identify the molecular mechanism by which deubiquitinating enzyme USP35 regulates cell cycle progression for the first time. Methods: USP35 RNAi was mainly used to identify the function of USP35 in cell cycle progression. To find substrates of USP35, we analyzed protein-protein interaction using LC-MS. Several biological methods, such as ubiquitination assay, cell synchronization, immunofluorescence, and immunoprecipitation assay were used to investigate the exact mechanism by which USP35 affects successful completion of mitosis. Results: USP35 knockdown caused not only reduction of mitotic cell number but also induction of mitotic cells with abnormal spindle formation. Actually, cell proliferation was decreased by USP35 knockdown. Interestingly, we found that loss of USP35 decreased the stability and expression of Aurora B, a member of chromosomal passenger complex (CPC), and the phosphorylation of its substrate. Indeed, USP35 interacted with Aurora B and deubiquitinated it. In addition, USP35 knockdown induced abnormal localization of Aurora B in mitotic cells. Finally, CDH1-mediated ubiquitination of Aurora B level was rescued by USP35 overexpression, but not inactive form of USP35, USP35 C450A. Discussion: Our findings suggest that USP35 regulates Aurora B-mediated mitotic spindle assembly and G2-M transition by blocking CDH1-induced degradation of Aurora B.

Keywords: USP35, HSP90, Aurora B, cell cycle progression

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Authors: Hanen Bouaziz, Françoise Croute, Najiba Zeghal

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In order to investigate the toxic effects of fluoride on cerebrum maturity of suckling mice, we treated adult female mice of Swiss Albinos strain by 500 ppm NaF in their drinking water from the 15th day of pregnancy until the day 14 after delivery. All mice were sacrificed on day 14 after parturition. During treatment, levels of thiobarbituric acid reactive substances, the marker of lipid peroxidation extend, increased, while the activities of the antioxidant enzymes such as glutathione peroxidase, superoxide dismutase and catalase and the level of glutathione decreased significantly in cerebellum compared with those of the control group. These results suggested that fluoride enhanced oxidative stress, thereby disturbing the antioxidant defense of nursing pups. In addition, acetylcholinesterase activity in cerebellum was inhibited after treatment with fluoride. In cerebellum of mice, migration of neurons from the external granular layer to the internal granular layer occurred postnatally. Key guidance signals to these migrating neurons were provided by laminin, an extracellular matrix protein fixed to the surface of astrocytes. In the present study, we examined the expression and distribution of laminin in cerebellum of 14-day-old mice. Immunoreactive laminin was disappeared by postnatal day 14 in cerebellum parenchyma of control pups and was restricted to vasculature despite the continued presence of granular cells in the external granular layer. In contrast, in cerebellum of NaF treated pups, laminin was deposited in organised punctuate clusters in the molecular layer. These data indicated that the disruption of laminin distribution might play a major role in the profound derangement of neuronal migration observed in cerebellum of NaF treated pups.

Keywords: acetylcholinesterase activity, cerebellum, laminin, oxidative stress, suckling mice

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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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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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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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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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Authors: Idan Chiyanzu, Maruping Mangena

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Crude glycerol, a major by-product from the transesterification of triacylglycerides with alcohol to biodiesel, is known to have a broad range of applications. For example, its bioconversion can afford a wide range of chemicals including alcohols, organic acids, hydrogen, solvents and intermediate compounds. In bacteria, the 2-oxoglutarate dioxygenase (2-OGD) enzymes are widely found among the Pseudomonas syringae species and have been recognized with an emerging importance in ethylene formation. However, the use of optimized enzyme function in recombinant systems for crude glycerol conversion to ethylene is still not been reported. The present study investigated the production of ethylene from crude glycerol using engineered E. coli MG1655 and JM109 strains. Ethylene production with an optimized expression system for 2-OGD in E. coli using a codon optimized construct of the ethylene-forming gene was studied. The codon-optimization resulted in a 20-fold increase of protein production and thus an enhanced production of the ethylene gas. For a reliable bioreactor performance, the effect of temperature, fermentation time, pH, substrate concentration, the concentration of methanol, concentration of potassium hydroxide and media supplements on ethylene yield was investigated. The results demonstrate that the recombinant enzyme can be used for future studies to exploit the conversion of low-priced crude glycerol into advanced value products like light olefins, and tools including recombineering techniques for DNA, molecular biology, and bioengineering can be used to allowing unlimited the production of ethylene directly from the fermentation of crude glycerol. It can be concluded that recombinant E.coli production systems represent significantly secure, renewable and environmentally safe alternative to thermochemical approach to ethylene production.

Keywords: crude glycerol, bioethylene, recombinant E. coli, optimization

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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

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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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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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Authors: Bryony Davies

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Many cell and tissue culture practices ignore the effects of gravity on cell biology, and little is known about how cell components may move in response to gravitational forces. Starfish oocytes provide an excellent model for interrogating the movement of cell components due to their unusually large size, ease of handling, and high transparency. Chromosomes from starfish oocytes can be visualised by microinjection of the histone-H2B-mCherry plasmid into the oocytes. The movement of the chromosomes can then be tracked by live-cell fluorescence microscopy. The results from experiments using these methods suggest that there is a replicable downward movement of centrally located chromosomes at a median velocity of 0.39 μm/min. Chromosomes nearer the nuclear boundary showed more restricted movement. Chromosome density and shape could also be altered by microinjection of restriction enzymes, primarily Alu1, before imaging. This was found to alter the speed of chromosome movement, with chromosomes from Alu1-injected nuclei showing a median downward velocity of 0.60 μm/min. Overall, these results suggest that there is a non-negligible movement of chromosomes in response to gravitational forces and that this movement can be altered by enzyme activity. Future directions based on these results could interrogate if this observed downward movement extends to other cell components and to other cell types. Additionally, it may be important to understand whether gravitational orientation and vertical positioning of cell components alter cell behaviour. The findings here may have implications for current cell culture practices, which do not replicate cell orientations or external forces experienced in vivo. It is possible that a failure to account for gravitational forces in 2D cell culture alters experimental results and the accuracy of conclusions drawn from them. Understanding possible behavioural changes in cells due to the effects of gravity would therefore be beneficial.

Keywords: starfish, oocytes, live-cell imaging, microinjection, chromosome dynamics

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Authors: Ronald Villamar-Torres, Michael Staudt, Christopher Viot

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Cotton Gossypium hirsutum L. is one of the most important industrial crops, producing the world leading natural textile fiber, but is very prone to arthropod attacks that reduce crop yield and quality. Cotton cultivation, therefore, makes an outstanding use of chemical pesticides. In reaction to herbivorous arthropods, cotton plants nevertheless show natural defense reactions, in particular through volatile organic compounds (VOCs) emissions. These natural defense mechanisms are nowadays underutilized but have a very high potential for cotton cultivation, and elucidating their genetic bases will help to improve their use. Simulating herbivory attacks by mechanical wounding of cotton plants in greenhouse, we studied by qPCR the changes in gene expression for genes of the terpenoids biosynthesis pathway. Differentially expressed genes corresponded to higher levels of the terpenoids biosynthesis pathway and not to enzymes synthesizing particular terpenoids. The genes were mapped on the G. hirsutum L. reference genome; their global relationships inside the general metabolic pathways and the biosynthesis of secondary metabolites were visualized with iPath2. The chromatographic profiles of VOCs emissions indicated first monoterpenes and sesquiterpenes emissions, dominantly four molecules known to be involved in plant reactions to arthropod attacks. As a result, the study permitted to identify potential key genes for the emission of volatile terpenoids by cotton plants in reaction to an arthropod attack, opening possibilities for molecular-assisted cotton breeding in benefit of smallholder cotton growers.

Keywords: biosynthesis pathways, cotton, mechanisms of plant defense, terpenoids, volatile organic compounds

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Authors: Olga Shapovalova, Vladimir Vinogradov

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Among many different sol – gel matrices only alumina can be successfully parenteral injected in the human body. And this is not surprising, because boehmite (aluminium oxyhydroxide) is the metal oxide approved by FDA and EMA for intravenous and intramuscular administrations, and also has been using for a longtime as adjuvant for producing of many modern vaccines. In our earlier study, it has been shown, that denaturation temperature of enzymes entrapped in sol-gel boehmite matrix increases for 30 – 60 °С with preserving of initial activity. It makes such matrices more attractive for long-term storage of non-stable drugs. In current work we present ultrasound-assisted sol-gel synthesis of nano-boehmite. This method provides bio-friendly, very stable, highly homogeneous alumina sol with using only water and aluminium isopropoxide as a precursor. Many parameters of the synthesis were studied in details: time of ultrasound treatment, US frequency, surface area, pore and nanoparticle size, zeta potential and others. Here we investigated the dependence of stability of colloidal sols and textural properties of the final composites as a function of the time of ultrasonic treatment. Chosen ultrasonic treatment time was between 30 and 180 minutes. Surface area, average pore diameter and total pore volume of the final composites were measured by surface and pore size analyzer Nova 1200 Quntachrome. It was shown that the matrices with ultrasonic treatment time equal to 90 minutes have the biggest surface area 431 ± 24 m2/g. On the other had such matrices have a smaller stability in comparison with the samples with ultrasonic treatment time equal to 120 minutes that have the surface area 390 ± 21 m2/g. It was shown that the stable sols could be formed only after 120 minutes of ultrasonic treatment, otherwise the white precipitate of boehmite is formed. We conclude that the optimal ultrasonic treatment time is 120 minutes.

Keywords: boehmite matrix, stabilisation, ultrasound-assisted sol-gel synthesis

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Authors: Kidane workelul, Li xu, Xiaoyang Pang, Jiaping Lv

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Dairy products are exceptionally ideal media for the growth of microorganisms because of their high nutritional content. There are several ways that milk might get contaminated throughout the milking process, including how the raw milk is transported and stored, as well as how long it is kept before being processed. Psychrotrophic bacteria are among the one which can deteriorate the quality of milk mainly their heat resistance proteas and lipase enzyme. For this research purpose 8 selected strains of Psychrotrophic bacteria (Entrococcus hirae, Pseudomonas fluorescens, Pseudomonas azotoformans, Pseudomonas putida, Exiguobacterium indicum, Pseudomonas paralactice, Acinetobacter indicum, Serratia liquefacients)are chosen and try to determine their characteristics based on the research methodology protocol. Thus, the 8 selected strains are cultured, plated incubate, extracted their genomic DNA and genome DNA was amplified, the purpose of the study was to identify their Psychrotrophic properties, lipase hydrolysis positive test, their optimal incubation temperature, designed primer using the noble strain P,flourescens conserved region area in target with lipA gene, optimized primer specificity as well as sensitivity and PCR detection for lipase positive strains using the design primers. Based on the findings both the selected 8 strains isolated from stored raw milk are Psychrotrophic bacteria, 6 of the selected strains except the 2 strains are positive for lipase hydrolysis, their optimal temperature is 20 to 30 OC, the designed primer specificity is very accurate and amplifies for those strains only with lipase positive but could not amplify for the others. Thus, the result is promising and could help in detecting the Psychrotrophic bacteria producing heat resistance enzymes (lipase) at early stage before the milk is processed and this will safe production loss for the dairy industry.

Keywords: dairy industry, heat-resistant, lipA, milk, primer and psychrotrophic

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Authors: Tarek Elnimr, Rabab El-kelany

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Heavy metals are environmental contaminants that may pose long-term health risks. Unfortunately, the consequent implementation of preventive measures was generally delayed, causing important negative effects to the exposed populations. The objective of this study was to determine whether co-consumption of nutritional supplements as selenium and vitamin E would treat the hazardous effects of exposure to mercury, lead and cadmium. 108 workers (60 males and 48 females) were the subject of this study, their ages ranged from 19-63 years, (M = 29.5±10.12). They were working in lamp factory for an average of 0.5-40 years (M= 5.3±8.8). Twenty control subjects matched for age and gender were used for comparison. All workers were subjected to neuropsychiatric evaluation. General Health Questionnaire (GHQ-28) revealed that 44.4% were complaining of anxiety, 52.7% of depression, 41.6% of social dysfunction and 22.2% of somatic symptoms. Cognitive tests revealed that long-term memory was not affected significantly when compared with controls, while short term memory and perceptual ability were affected significantly. Blood metal levels were measured by Inductively Coupled Plasma – optical emission spectrometry(ICP-OES), and revealed that the mean blood mercury, lead and cadmium concentrations before treatment were 1.6 mg/l, 0.39 mg/l and 1.7 µg/l, while they decreased significantly after treatment to 1.2 mg/l, 0.29 mg/l and 1.3 µg/l respectively. Anti-oxidative enzymes (paraoxonase and catalase) and lipid peroxidation product (malondialdehyde) were measured before and after treatment with selenium and vitamin E, and showed significant improvement. It could be concluded that co-consumption of selenium and vitamin E produces significant decrease in mercury, lead and cadmium levels in blood.

Keywords: mercury, lead, cadmium, neuropsychiatric impairment, selenium, vitamin E

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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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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

Procedia PDF Downloads 88

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

Abstract:

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 (η_bend) rounding 90%, was reached.

Keywords: bendiocarb, biodegradation, biofilm reactor, carbamate insecticide

Procedia PDF Downloads 261

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 142

Authors: Peng Hou

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Natural products produced from marine bacteria serve as an immense reservoir for anti-infective drugs and therapeutic agents. Nowadays, heterologous expression of gene clusters of interests has been widely adopted as an effective strategy for natural product discovery. Briefly, the heterologous expression flowchart would be: biosynthetic gene cluster identification, pathway construction and expression, and product detection. However, gene cluster capture using traditional Transformation-associated recombination (TAR) protocol is low-efficient (0.5% positive colony rate). To make things worse, most of these putative new natural products are only predicted by bioinformatics analysis such as antiSMASH, and their corresponding natural products biosynthetic pathways are either not expressed or expressed at very low levels under laboratory conditions. Those setbacks have inspired us to focus on seeking new technologies to efficiently edit and refractor of biosynthetic gene clusters. Recently, two cutting-edge techniques have attracted our attention - the CRISPR-Cas9 and Gibson Assembly. By now, we have tried to pretreat Brevibacillus laterosporus strain genomic DNA with CRISPR-Cas9 nucleases that specifically generated breaks near the gene cluster of interest. This trial resulted in an increase in the efficiency of gene cluster capture (9%). Moreover, using Gibson Assembly by adding/deleting certain operon and tailoring enzymes regardless of end compatibility, the silent construct (~80kb) has been successfully refactored into an active one, yielded a series of analogs expected. With the appearances of the novel molecular tools, we are confident to believe that development of a high throughput mature pipeline for DNA assembly, transformation, product isolation and identification would no longer be a daydream for marine natural product discovery.

Keywords: biosynthesis, CRISPR-Cas9, DNA assembly, refactor, TAR cloning

Procedia PDF Downloads 268

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

Procedia PDF Downloads 193

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.

Procedia PDF Downloads 456

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 182

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

Procedia PDF Downloads 122