Search results for: bacteria isolation

Authors: Sang Guen Kim, Sib Sankar Giri, Jin Woo Jun, Saekil Yun, Hyoun Joong Kim, Sang Wha Kim, Jung Woo Kang, Se Jin Han, Se Chang Park

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Emerging of the super bacteria, bacteriophages are considered to be as an alternative to antibiotics. As the demand of phage increased, economical and large production of phage is becoming one of the critical points. For the therapeutic use, what is important is to eradicate the pathogenic bacteria as fast as possible, so higher concentration of phages is generally needed for effective therapeutic function. On the contrary, for the maximum production, bacteria work as a phage producing factory. As a microbial cell factory, bacteria is needed to last longer producing the phages without eradication. Consequently, killing the bacteria fast has a negative effect on large production. In this study, Multiplicity of Infection (MOI) was manipulated based on initial bacterial inoculation and used phage pAh-6C which has therapeutic effect against Aeromonas hydrophila. 1, 5 and 10 percent of overnight bacterial culture was inoculated and each bacterial culture was co-cultured with the phage of which MOI of 0.01, 0.0001, and 0.000001 respectively. Simply changing the initial MOI as well as bacterial inoculation concentration has regulated the production quantity of the phage without any other changes to culture conditions. It is anticipated that this result can be used as a foundational data for mass production of lytic bacteriophages which can be used as the therapeutic bio-control agent.

Keywords: bacteriophage, multiplicity of infection, optimization, Aeromonas hydrophila

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Authors: Adrien Pyskir, Manuel Collet, Zoran Dimitrijevic, Claude-Henri Lamarque

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During the last decades, great efforts have been made to reduce acoustic and vibrational disturbances in transportations, as it has become a key feature for comfort. Today, isolation and design have neutralized most of the troublesome vibrations, so that cars are quieter and more comfortable than ever. However, some problems remain unsolved, in particular concerning low-frequency isolation and the frequency-dependent stiffening of materials like rubber. To sum it up, a balance has to be found between a high static stiffness to sustain the vibration source’s mass, and low dynamic stiffness, as wideband as possible. Systems meeting these criteria are yet to be designed. We thus investigated solutions inspired by metamaterials to control efficiently low-frequency wave propagation. Structures exhibiting a negative Poisson ratio, also called auxetic structures, are known to influence the propagation of waves through beaming or damping. However, their stiffness can be quite peculiar as well, as they can present regions of zero stiffness on the stress-strain curve for compression. In addition, auxetic materials can be easily adapted in many ways, inducing great tuning potential. Using finite element software COMSOL Multiphysics, a resonant design has been tested through statics and dynamics simulations. These results are compared to experimental results. In particular, the bandgaps featured by these structures are analyzed as a function of design parameters. Great stiffness properties can be observed, including low-frequency dynamic stiffness loss and broadband transmission loss. Such features are very promising for practical isolation purpose, and we hope to adopt this kind of metamaterial into an effective industrial damper.

Keywords: auxetics, metamaterials, structural dynamics, vibration isolation

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Authors: A. Zh. Aupova, A. Ulankyzy, A. Sarsenova, A. Kussayin, Sh. Turarbek, N. Moldagulova, A. Kurmanbayev

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One of the methods of organic waste bioconversion into environmentally-friendly fertilizer is composting. Microorganisms that produce hydrolytic enzymes play a significant role in accelerating the process of organic waste composting. We studied the enzymatic potential (amylase, protease, cellulase, lipase, urease activity) of bacteria isolated from the sewage sludge of Nur-Sultan, Rudny, and Fort-Shevchenko cities, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha for processing organic waste and identifying active strains. Microorganism isolation was carried out by the cultures enrichment method on liquid nutrient media, followed by inoculating on different solid media to isolate individual colonies. As a result, sixty-one microorganisms were isolated, three of which were thermophiles (DS1, DS2, and DS3). The highest number of isolates, twenty-one and eighteen, were isolated from sewage sludge of Nur-Sultan and Rudny cities, respectively. Ten isolates were isolated from the wastewater of the sewage treatment plant in Fort-Shevchenko. From the dacha soil of Nur-Sultan city and freshly cut grass - 9 and 5 isolates were revealed, respectively. The lipolytic, proteolytic, amylolytic, cellulolytic, ureolytic, and oil-oxidizing activities of isolates were studied. According to the results of experiments, starch hydrolysis (amylolytic activity) was found in 2 isolates - CB2/2, and CB2/1. Three isolates - CB2, CB2/1, and CB1/1 were selected for the highest ability to break down casein. Among isolated 61 bacterial cultures, three isolates could break down fats - CB3, CBG1/1, and IL3. Seven strains had cellulolytic activity - DS1, DS2, IL3, IL5, P2, P5, and P3. Six isolates rapidly decomposed urea. Isolate P1 could break down casein and cellulose. Isolate DS3 was a thermophile and had cellulolytic activity. Thus, based on the conducted studies, 15 isolates were selected as a potential for sewage sludge composting - CB2, CB3, CB1/1, CB2/2, CBG1/1, CB2/1, DS1, DS2, DS3, IL3, IL5, P1, P2, P5, P3. Selected strains were identified on a mass spectrometer (Maldi-TOF). The isolate - CB 3 was referred to the genus Rhodococcus rhodochrous; two isolates CB2 and CB1 / 1 - to Bacillus cereus, CB 2/2 - to Cryseobacterium arachidis, CBG 1/1 - to Pseudoxanthomonas sp., CB2/1 - to Bacillus megaterium, DS1 - to Pediococcus acidilactici, DS2 - to Paenibacillus residui, DS3 - to Brevibacillus invocatus, three strains IL3, P5, P3 - to Enterobacter cloacae, two strains IL5, P2 - to Ochrobactrum intermedium, and P1 - Bacillus lichenoformis. Hence, 60 isolates were isolated from the wastewater of the cities of Nur-Sultan, Rudny, Fort-Shevchenko, the dacha soil of Nur-Sultan city, and freshly cut grass from the dacha. Based on the highest enzymatic activity, 15 active isolates were selected and identified. These strains may become the candidates for bio preparation for sewage sludge processing.

Keywords: sewage sludge, composting, bacteria, enzymatic activity

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Authors: Omneya M. Helmy, Mona T. Kashef

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Aminoglycosides are used in treating a wide range of infections caused by both Gram-negative and Gram positive bacteria. The presence of 16S rRNA methyl transferases (16S-RMTase) is among the newly discovered resistance mechanisms that confer high resistance to clinically useful aminoglycosides. Cephalosporins are the most commonly used antimicrobials in Egypt; therefore, this study was conducted to determine the isolation frequency of 16S rRNA methyl transferases among third generation cephalosporin-resistant clinical isolates in Egypt. One hundred and twenty three cephalosporin resistant Gram-negative clinical isolates were screened for aminoglycoside resistance by the Kirby Bauer disk diffusion method and tested for possible production of 16S-RMTase. PCR testing and sequencing were used to confirm the presence of 16S-RMTase and the associated antimicrobial resistance determinants, as well as the genetic region surrounding the armA gene. Out of 123 isolates, 66 (53.66%) were resistant to at least one aminoglycoside antibiotic. Only one Escherichia coli isolate (E9ECMO) which was totally resistant to all tested aminoglycosides, was confirmed to have the armA gene in association with blaTEM-1, blaCTX-M-15, blaCTX-M-14 and aac(6)-Ib genes. The armA gene was found to be carried on a large A/C plasmid. Genetic mapping of the armA surrounding region revealed, for the first time, the association of armA with aac(6)-Ib on the same transposon. In Conclusion, the isolation frequency of 16S-RMTase was low among the tested cephalosporin-resistant clinical samples. However, a novel composite transposon has been detected conferring high-level aminoglycosides resistance.

Keywords: aminoglcosides, armA gene, β lactmases, 16S rRNA methyl transferases

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Authors: Maria Nejjari, Michel Cloutier, Guylaine Talbot, Martin Lanthier

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The fluorescence in situ hybridization (FISH) is a staining technique that allows the identification, detection and quantification of microorganisms without prior cultivation by means of epifluorescence and confocal laser scanning microscopy (CLSM). Oligonucleotide probes have been used to detect bacteria and archaea that colonize the cattle and swine digestive systems. These bacterial strains have been obtained from fecal samples issued from cattle manure and swine slurry. The collection of these samples has been done at 3 different pit’s levels A, B and C with same height. Two collection depth levels have been taken in consideration, one collection level just under the pit’s surface and the second one at the bottom of the pit. Cells were fixed and FISH was performed using oligonucleotides of 15 to 25 nucleotides of length associated with a fluorescent molecule Cy3 or Cy5. The double hybridization using Cy3 probe targeting bacteria (Cy3-EUB338-I) along with a Cy5 probe targeting Archaea (Gy5-ARCH915) gave a better signal. The CLSM images show that there are more bacteria than archaea in swine slurry. However, the choice of fluorescent probes is critical for getting the double hybridization and a unique signature for each microorganism. FISH technique is an easy way to detect pathogens like E. coli O157, Listeria, Salmonella that easily contaminate water streams, agricultural soils and, consequently, food products and endanger human health.

Keywords: archaea, bacteria, detection, FISH, fluorescence

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Authors: Jerry John T. M., Sylas V. P., Shijo Joy

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Hydrogen is the most essential gas that can be used for many purposes. During the production of hydrogen using raw materials like Soil and leftover cooked rice water (kanjivellam), the major by-product formed is water. Soil is collected from three different places in kottayam district: Kallara, Meenachilar, and Athirampuzha. Collected samples are mixed with rice water and tested for traces of hydrogen using a biohydrogen sensor after 72 hours. The result was the presence of hydrogen in all the 3 samples. After streaking, PCR and gel electrophoresis detected the bacteria which produced the hydrogen. RGCB Thiruvananthapuram conducted the sequencing of the PCR resultant. And identified the bacterial strains. Five variants of Bacillus bacteria ( (1) Bacillus cereus strain JTM GenBank: OP278839.1 (2) Bacillus toyonensis strain JTM2 GenBank: OP278841.1 (3) Bacillus anthracis strain JTM_SR2989-3-R_H08 GenBank: OP278960.1 (4) Bacillus thuringiensis strain JRY1 GenBank: OP278976.1 (5) Bacillus anthracis strain JTM_SR2989-3-F_H07 GenBank: OP278959.1 ) are identified and successfully registered in NCBI Gen bank. These Bacillus bacteria are major types of Rhizobacteria that can form spores and can survive in the soil for a long time period under harsh environmental conditions. Also, plant growth is enhanced by PGPR (Plant growth promoting rhizobacteria) through the induction of systemic resistance, antibiosis, and competitive omission. The molecular sequencing was submitted to the NCBI Gen Bank, and the accession numbers were allotted for the bacterial cultures.

Keywords: bio hydrogen production, bacterial bio hydrogen production, plant related to bacillus bacteria., bacillus bacteria study

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Authors: Sugandha Asthana, Geetika Vajpayee, Pratibha Kumari, Shanthy Sundaram

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An economically important disease of wheat in North Western region of India is Karnal Bunt caused by smut fungus Tilletia indica. This fungal pathogen spreads by air, soil and seed borne sporodia at the time of flowering, which ultimately leads to partial bunting of wheat kernels with fishy odor and taste to wheat flour. It has very serious effects due to quarantine measures which have to be applied for grain exports. Chemical fungicides such as mercurial compounds and Propiconazole applied to the control of Karnal bunt have been only partially successful. Considering the harmful effects of chemical fungicides on man as well as environment, many countries are developing biological control as the superior substitute to chemical control. Repeated use of fungicides can be responsible for the development of resistance in fungal pathogens against certain chemical compounds. The present investigation is based on the isolation and evaluation of antifungal properties of some isolated (from natural manure) and commercial bacterial strains against Tilletia indica. Total 23 bacterial isolates were obtained and antagonistic activity of all isolates and commercial bacterial strains (Bacillus subtilis MTCC8601, Bacillus pumilus MTCC 8743, Pseudomonas aeruginosa) were tested against T. indica by dual culture plate assay (pour plate and streak plate). Test for the production of antifungal volatile organic compounds (VOCs) by antagonistic bacteria was done by sealed plate method. Amongst all s1, s3, s5, and B. subtilis showed more than 80% inhibition. Production of extracellular hydrolytic enzymes such as protease, beta 1, 4 glucanase, HCN and ammonia was studied for confirmation of antifungal activity. s1, s3, s5 and B. subtilis were found to be the best for protease activity and s5 and B. subtilis for beta 1, 4 glucanase activity. Bacillus subtilis was significantly effective for HCN whereas s3, s5 and Bacillus subtilis for ammonia production. Isolates were identified as Pseudomonas aeruginosa (s1) and B. licheniformis (s3, s5) by various biochemical assays and confirmed by16s rRNA sequencing. Use of microorganisms or their secretions as biocontrol agents to avoid plant diseases is ecologically safe and may offer long term of protection to crop. The above study reports the promising effects of these strains in better pathogen free crop production and quality maintenance as well as prevention of the excessive use of synthetic fungicides.

Keywords: antagonistic, antifungal, biocontrol, Karnal bunt

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Authors: Elmo Thiao Lins Cöuras Ford, Valentina Alessandra Carvalho do Vale, R. M. Nascimento, J. U. L. Mendes

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Now the great majority of the applications of thermal isolation in the strip of drops and averages temperatures (up to 200ºC), it is made being used from aggressive materials to the nature such an as: glass wool, rock wool, polystyrene, EPS among others. Such materials, in spite of the effectiveness in the retention of the flow of heat, possess considerable cost and when discarded they are long years to be to decompose. In that context, trying to adapt the world politics the about of the preservation of the environment, a study began with intention of developing a material composite, with properties of thermal, originating from insulating industrial residues. In this research, the behavior of the composite was analyzed, as submitted the fire. For this, the reaction rehearsals were accomplished to the fire for the composites 2:1; 1:1; 1:2 and for the latex, based in the "con" experiment in agreement with the norm ASTM–E 1334-90. As consequence, in function of the answers of the system, was possible to observe to the acting of each mixture proportion.

Keywords: composite, Latex, reacion to the fire, thermal isolation

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Authors: Shashank Gahlaut, Chandrashekhar Sharan, J. P. Singh

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Increasing incidence of antibiotic-resistant bacteria is a big concern for the treatment of pathogenic diseases. The effect of treatment of patients with antibiotics often leads to the evolution of antibiotic resistance in the pathogens. The detection of antibiotic or antimicrobial resistant bacteria (microbes) is quite essential as it is becoming one of the big threats globally. Here we propose a novel technique to tackle this problem. We are taking a step forward to prevent the infections and diseases due to drug resistant microbes. This detection is based on some unique features of silver (a noble metal) nanorods (AgNRs) which are fabricated by a physical deposition method called thermal glancing angle deposition (GLAD). Silver nanorods are found to be highly sensitive and selective for hydrogen sulfide (H2S) gas. Color and water wetting (contact angle) of AgNRs are two parameters what are effected in the presence of this gas. H₂S is one of the major gaseous products evolved in the bacterial metabolic process. It is also known as gasotransmitter that transmits some biological singles in living systems. Nitric Oxide (NO) and Carbon mono oxide (CO) are two another members of this family. Orlowski (1895) observed the emission of H₂S by the bacteria for the first time. Most of the microorganism produce these gases. Here we are focusing on H₂S gas evolution to determine live/dead and antibiotic-resistant bacteria. AgNRs array has been used for the detection of H₂S from micro-organisms. A mobile app is also developed to make it easy, portable, user-friendly, and cost-effective.

Keywords: antibiotic resistance, hydrogen sulfide, live and dead bacteria, mobile app

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Authors: Ahmad Akrad, Rabia Sehab, Fadi Alyoussef

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Nowadays, induction machines are widely used in industry thankful to their advantages comparing to other technologies. Indeed, there is a big demand because of their reliability, robustness and cost. The objective of this paper is to deal with diagnosis, detection and isolation of faults in a three-phase induction machine. Among the faults, Inter-turn short-circuit fault (ITSC), current sensors fault and single-phase open circuit fault are selected to deal with. However, a fault detection method is suggested using residual errors generated by the root mean square (RMS) of phase currents. The application of this method is based on an asymmetric nonlinear model of Induction Machine considering the winding fault of the three axes frame state space. In addition, current sensor redundancy and sensor fault detection and isolation (FDI) are adopted to ensure safety operation of induction machine drive. Finally, a validation is carried out by simulation in healthy and faulty operation modes to show the benefit of the proposed method to detect and to locate with, a high reliability, the three types of faults.

Keywords: induction machine, asymmetric nonlinear model, fault diagnosis, inter-turn short-circuit fault, root mean square, current sensor fault, fault detection and isolation

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Authors: Yo-Sheng Lin, Jen-How Lee, Chien-Chin Wang

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A 60-GHz receiver front-end using standard 90-nm CMOS technology is reported. The receiver front-end comprises a wideband low-noise amplifier (LNA), and a double-balanced Gilbert cell mixer with a current-reused RF single-to-differential (STD) converter, an LO Marchand balun and a baseband amplifier. The receiver front-end consumes 34.4 mW and achieves LO-RF isolation of 60.7 dB, LO-IF isolation of 45.3 dB and RF-IF isolation of 41.9 dB at RF of 60 GHz and LO of 59.9 GHz. At IF of 0.1 GHz, the receiver front-end achieves maximum conversion gain (CG) of 26.1 dB at RF of 64 GHz and CG of 25.2 dB at RF of 60 GHz. The corresponding 3-dB bandwidth of RF is 7.3 GHz (58.4 GHz to 65.7 GHz). The measured minimum noise figure was 5.6 dB at 64 GHz, one of the best results ever reported for a 60 GHz CMOS receiver front-end. In addition, the measured input 1-dB compression point and input third-order inter-modulation point are -33.1 dBm and -23.3 dBm, respectively, at 60 GHz. These results demonstrate the proposed receiver front-end architecture is very promising for 60 GHz direct-conversion transceiver applications.

Keywords: CMOS, 60 GHz, direct-conversion transceiver, LNA, down-conversion mixer, marchand balun, current-reused

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Authors: Shahrokh Barati, Reza Ramezani

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Due to the countries growing attention to the renewable energy producing, the demand for energy from renewable energy has gone up among the renewable energy sources; wind energy is the fastest growth in recent years. In this regard, in order to increase the availability of wind turbines, using of Fault Detection and Isolation (FDI) system is necessary. Wind turbines include of various faults such as sensors fault, actuator faults, network connection fault, mechanical faults and faults in the generator subsystem. Although, sensors and actuators have a large number of faults in wind turbine but have discussed fewer in the literature. Therefore, in this work, we focus our attention to design a sensor and actuator fault detection and isolation algorithm and Fault-tolerant control systems (FTCS) for Wind Turbine. The aim of this research is to propose a comprehensive fault detection and isolation system for sensors and actuators of wind turbine based on data-driven approaches. To achieve this goal, the features of measurable signals in real wind turbine extract in any condition. The next step is the feature selection among the extract in any condition. The next step is the feature selection among the extracted features. Features are selected that led to maximum separation networks that implemented in parallel and results of classifiers fused together. In order to maximize the reliability of decision on fault, the property of fault repeatability is used.

Keywords: FDI, wind turbines, sensors and actuators faults, renewable energy

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Authors: Abdulrahman Abdulhamid Arabo, Raji Arabi Bamanga, Mujiburrahman Fadilu, Musa Abubakar, Fatima Abdullahi Shehu, Hafeez Muhammad Yakasai, Nasiru Abdullahi

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The aim of this study was to isolate and identify biosurfactant-producing and diesel alkanes degrading bacteria. For this reason, bacteria isolated from the diesel-contaminated site were screened for their potential to produce biosurfactants and degrade diesel alkanes. Primary selection of diesel degraders was carried out by using the conventional enrichment culture technique, where 12 bacterial strains were isolated based on their ability to grow on minimal media supplemented with diesel as the sole carbon source, which was followed by qualitative screening methods for potential biosurfactant production. Isolate B11 was the only candidate that showed positive signs for drop collapse, foaming, hemolytic test, oil displacement of more than 22 ± 0.05 mm, and emulsification (E24) of 14 ± 0.30%. The effect of various culture parameters (incubation time, diesel concentration, nitrogen source, pH and temperature) on the biodegradation of diesel was evaluated. The optimum incubation time was confirmed to be 120 days for isolate B11, and the optimum PH was confirmed as 8.0 for the isolate; similarly, the optimum temperature was confirmed as 35oC. In addition, diesel oil was used as the sole carbon source for the isolates. The favorable diesel concentration was 12.5 % (v/v) for the isolate. The isolate has shown degradative ability towards Tridecane (C13), dodecane, 2, 6, 10-trimethyl- (C15), Tetradecane (C14), 2,6,10-Trimethyltridecane (C16), Pentadecane (C15). It degraded between 0.27% - 9.65% of individual diesel oil alkanes. The strain has exhibited the potential of degrading diesel oil n-alkanes and was identified as Alcaligenes species strain B11 (MZ027604) using the 16S rRNA. Sequencing.

Keywords: diesel oil, biosurfactant, Alcaligenes sp, biodegradation

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Authors: Sumeyra Sevim, Gulsum Gizem Topal, Mercan Merve Tengilimoglu-Metin, Mevlude Kizil

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Aflatoxin M1 (AFM1) is a major toxic and carcinogenic molecule in milk and milk products. Therefore, it poses a risk for public health. Probiotics can be biological agent to remove AFM1. The aim of this study is to evaluate the effect of probiotic bacteria on AFM1 detoxification in phosphate buffer saline. The PBS samples artificially contaminated with AFM1 at concentration 100 pg/ml were prepared with probiotics bacteria that including monoculture (L. plantarum, B. bifidum ATCC, B. animalis ATCC 27672) and binary culture (L. bulgaricus + S. thermophiles, B. bifidum ATCC + B. animalis ATCC 27672, L. plantarum+B. bifidum ATCC, L. plantarum+ B. animalis ATCC 27672). The samples were incubated at 37°C for 4 hours and stored for 1, 5 and 10 days. The toxin was measured by the ELISA. The highest levels of AFM1 binding ability (63.6%) in PBS were detected yoghurt starter bacteria, while L. plantarum had the lowest levels of AFM1 binding ability (35.5%) in PBS. In addition, it was found that there was significant effect of storage on AFM1 binding ability in all groups except the one including B. animalis (p < 0.05). Consequently, results demonstrate that AFM1 detoxification by probiotic bacteria has a potential application to reduce toxin concentrations in yoghurt. Moreover, probiotic strains can react with itself as synergic or antagonist.

Keywords: aflatoxin M1, ELISA, probiotics, storage

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Authors: Sulaiman B. Ali Alharbi, Bassam H. Mashat, Naif Abdullah Al-Harbi, Milton Wainwright, Abeer S. Aloufi, Sulamain Alnaimat

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Bismuth salicylate was found to inhibit the growth of a range of bacteria and yeast, Candida albican. In general the growth of bacteria did not result in the increase in bismuth solubilisation, in contrast, bismuth solubilisation increased following the growth of C. albicans. A significant increase in the biomass (dry weight) of Aspergillus niger and Aspergillus oryzae occurred in vitro when these fungi were grown in the presence of bismuth salicylate. Biomass increase occurred over a range of bismuth compound additions, which in the case of A. oryzae was associated with the increase in the solubilisation of the insoluble bismuth compounds.

Keywords: bacterial inhibition, fungal growth stimulation, medical uses of bismuth, yeast inhibition

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Authors: A. T. Kuda, J. J. Dayya, A. Jimoh

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This paper investigates the fault detection and Isolation technique of measurement data sets from a three tank system using analytical model-based temporal redundancy which is based on residual generation using parity equations/space approach. It further briefly outlines other approaches of model-based residual generation. The basic idea of parity space residual generation in temporal redundancy is dynamic relationship between sensor outputs and actuator inputs (input-output model). These residuals where then used to detect whether or not the system is faulty and indicate the location of the fault when it is faulty. The method obtains good results by detecting and isolating faults from the considered data sets measurements generated from the system.

Keywords: fault detection, fault isolation, disturbing influences, system failure, parity equation/relation, structured parity equations

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Authors: Priyanka Sharma, Ranjita Das, Mohan C. Kalita, Debajit Thakur

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Background: Actinomycetes are the richest source of novel bioactive secondary metabolites such as antibiotics, enzymes and other therapeutically useful metabolites with diverse biological activities. The present study aims at the antimicrobial potential and genetic diversity of culturable Actinomycetes isolated from protected forest ecosystems of Assam which includes Kaziranga National Park (26°30˝-26°45˝N and 93°08˝-93°36˝E), Pobitora Wildlife Sanctuary (26º12˝-26º16˝N and 91º58˝-92º05˝E) and Gibbon Wildlife Sanctuary (26˚40˝-26˚45˝N and 94˚20˝-94˚25˝E) which are located in the North-eastern part of India. Northeast India is a part of the Indo-Burma mega biodiversity hotspot and most of the protected forests of this region are still unexplored for the isolation of effective antibiotic-producing Actinomycetes. Thus, there is tremendous possibility that these virgin forests could be a potential storehouse of novel microorganisms, particularly Actinomycetes, exhibiting diverse biological properties. Methodology: Soil samples were collected from different ecological niches of the protected forest ecosystems of Assam and Actinomycetes were isolated by serial dilution spread plate technique using five selective isolation media. Preliminary screening of Actinomycetes for an antimicrobial activity was done by spot inoculation method and the secondary screening by disc diffusion method against several test pathogens, including multidrug resistant Staphylococcus aureus (MRSA). The strains were further screened for the presence of antibiotic synthetic genes such as type I polyketide synthases (PKS-I), type II polyketide synthases (PKS-II) and non-ribosomal peptide synthetases (NRPS) genes. Genetic diversity of the Actinomycetes producing antimicrobial metabolites was analyzed through 16S rDNA-RFLP using Hinf1 restriction endonuclease. Results: Based on the phenotypic characterization, a total of 172 morphologically distinct Actinomycetes were isolated and screened for antimicrobial activity by spot inoculation method on agar medium. Among the strains tested, 102 (59.3%) strains showed activity against Gram-positive bacteria, 98 (56.97%) against Gram-negative bacteria, 92 (53.48%) against Candida albicans MTCC 227 and 130 (75.58%) strains showed activity against at least one of the test pathogens. Twelve Actinomycetes exhibited broad spectrum antimicrobial activity in the secondary screening. The taxonomic identification of these twelve strains by 16S rDNA sequencing revealed that Streptomyces was found to be the predominant genus. The PKS-I, PKS-II and NRPS genes detection indicated diverse bioactive products of these twelve Actinomycetes. Genetic diversity by 16S rDNA-RFLP indicated that Streptomyces was the dominant genus amongst the antimicrobial metabolite producing Actinomycetes. Conclusion: These findings imply that Actinomycetes from the protected forest ecosystems of Assam, India, are a potential source of bioactive secondary metabolites. These areas are as yet poorly studied and represent diverse and largely unscreened ecosystem for the isolation of potent Actinomycetes producing antimicrobial secondary metabolites. Detailed characterization of the bioactive Actinomycetes as well as purification and structure elucidation of the bioactive compounds from the potent Actinomycetes is the subject of ongoing investigation. Thus, to exploit Actinomycetes from such unexplored forest ecosystems is a way to develop bioactive products.

Keywords: Actinomycetes, antimicrobial activity, forest ecosystems, RFLP

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Authors: Omar Alhamd, Peter A. Thomas, Trevor J. Greenhough, Annette K. Shrive

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The Pseudomonas syringae species complex includes many plant pathogenic strains with highly specific interactions with varied host species and cultivars. The rapid spread of these bacteria over the last ten years has become a cause for concern. Nanoparticles have previously shown promise in microbiological action. We have therefore investigated in vitro and in vivo the effects of different types and sizes of nanoparticles in order to provide quantitative information about their effect on the bacteria. The effects of several different nanoparticles against several bacteria strains were investigated. The effect of NP on bacterial growth was studied by measuring the optical density, biochemical and nutritional tests, and transmission electron microscopy (TEM) to determine the shape and size of NP. Our results indicate that their effects varied, with either a negative or a positive impact on both bacterial and plant growth. Additionally, the methods of exposure to nanoparticles have a crucial role in accumulation, translocation, growth response and bacterial growth. The results of our studies on the behaviour and effects of nanoparticles in model plants showed. Cerium oxide (CeO₂) and silver (Ag) NP showed significant antibacterial activity against several pathogenic bacteria. It was found that titanium nanoparticles (TiO₂) can have either a negative or a positive impact, according to concentration and size. It is also thought that environmental conditions can have a major influence on bacterial growth. Studies were therefore also carried out under some environmental stress conditions to test bacterial survival and to assess bacterial virulence. All results will be presented including information about the effects of different nanoparticles on Pseudomonas syringae bacteria.

Keywords: plant microbiome, nanoparticles, 16S rRNA gene sequencing, bacterial survival

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Authors: P. Behera, K. K. Singh, D. K. Saini, M. De

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Implementation of 2D materials in the detection of bio analytes is highly advantageous in the field of sensing because of its high surface to volume ratio. We have designed our sensor array with different cationic two-dimensional MoS₂, where surface modification was achieved by cationic thiol ligands with different functionality. Green fluorescent protein (GFP) was chosen as signal transducers for its biocompatibility and anionic nature, which can bind to the cationic MoS₂ surface easily, followed by fluorescence quenching. The addition of bio-analyte to the sensor can decomplex the cationic MoS₂ and GFP conjugates, followed by the regeneration of GFP fluorescence. The fluorescence response pattern belongs to various analytes collected and transformed to linear discriminant analysis (LDA) for classification. At first, 15 different proteins having wide range of molecular weight and isoelectric points were successfully discriminated at 50 nM with detection limit of 1 nM. The sensor system was also executed in biofluids such as serum, where 10 different proteins at 2.5 μM were well separated. After successful discrimination of protein analytes, the sensor array was implemented for bacteria sensing. Six different bacteria were successfully classified at OD = 0.05 with a detection limit corresponding to OD = 0.005. The optimized sensor array was able to classify uropathogens from non-uropathogens in urine medium. Further, the technique was applied for discrimination of bacteria possessing resistance to different types and amounts of drugs. We found out the mechanism of sensing through optical and electrodynamic studies, which indicates the interaction between bacteria with the sensor system was mainly due to electrostatic force of interactions, but the separation of native bacteria from their drug resistant variant was due to Van der Waals forces. There are two ways bacteria can be detected, i.e., through bacterial cells and lysates. The bacterial lysates contain intracellular information and also safe to analysis as it does not contain live cells. Lysates of different drug resistant bacteria were patterned effectively from the native strain. From unknown sample analysis, we found that discrimination of bacterial cells is more sensitive than that of lysates. But the analyst can prefer bacterial lysates over live cells for safer analysis.

Keywords: array-based sensing, drug resistant bacteria, linear discriminant analysis, two-dimensional MoS₂

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Authors: Cho Achidi

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Tomato is a crucial food crop significantly contributes to global food and nutrition security. However, postharvest losses severely limit its role. Therefore, it is necessary to develop sustainable strategies to minimize these losses and improve the shelf-life of tomato fruits. One of the major concerns is bacterial infections, particularly by faecal coliform bacteria, which can cause food poisoning and illnesses like diarrhoea and dysentery. This study seeks to identify the presence of coliform bacteria on tomato fruits in fields and markets in Muea, Buea Municipality. The study also evaluated different management strategies to reduce the bacterial incidence and load on tomato fruits. A total of 200 fruits were sampled for both the coliform survey and shelf-life analysis. Ten farmers and traders provided samples, including asymptomatic and symptomatic tomato fruits. The samples designated for shelf-life analysis were treated with Aquatab, warm water, lemon, and onion. The results indicated that out of the 80 symptomatic samples collected, 12.5% contained faecal and total coliform species. Among the ten farms sampled, 14% were infected with coliform bacteria, with the highest infestation rate of 60% recorded in field 4. Furthermore, 15% of the asymptomatic tomato fruits were found to be infected by coliform bacteria. Regarding the management strategies, Aquatabs exhibited the highest efficacy in reducing the incidence of coliform bacteria on tomato fruits, followed by onion and lemon extracts. Although hot water treatment effectively removed bacteria from the fruits, damaging the cell wall negatively affected their shelf-life. Overall, this study emphasizes the severity of coliform bacterial pathogens in the Muea area, particularly their occurrence on asymptomatic tomatoes, which poses a significant concern for plant quarantine services. It also demonstrates potential options for mitigating this bacterial challenge.

Keywords: tomato, shelf-life analysis, food and nutrition security, coliform bbacteria

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Authors: Kusumawadee Thancharoen, Rungrat Nontawong, Thanawat Junsom

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Pathogenic bacterial flora was isolated from giant freshwater prawns, Macrobrachium rosenbergii. Infected shrimp samples were collected from BuaBan Aquafarm in Kalasin Province, Thailand, between June and September 2018. Bacterial species were isolated by serial dilution and plated on Thiosulfate Citrate Bile Salt Sucrose (TCBS) agar medium. A total 89 colonies were isolated and identified using the API 20E biochemical tests. Results showed the presence of genera Aeromonas, Citrobacter, Chromobacterium, Providencia, Pseudomonas, Stenotrophomonas and Vibrio. Maximum number of species was recorded in Pseudomonas (50.57%) with minimum observed in Chromobacterium and Providencia (1.12%).

Keywords: biochemical test, giant freshwater prawn, isolation, salt tolerance, shrimp diseases

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Authors: Jaruwan Chutrtong

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

Keywords: dynamic, dry yoghurt, storage, temperature

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Authors: Siewchuiang Sia, Gimcheong Tan

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Bacteriophages are the most abundant and genetically diverse living entities on earth; they help in regulating and maintaining microbial diversity and balance in its natural ecosystem. In this study, the infectivity of SFN6B tailed phage was investigated in various water samples. Host bacteria (Shigella flexneri) were spiked in sterilized environmental and domestic water samples, followed by SFN6B treatment. Two incubation conditions were selected for this study, 37 oC and room temperature. S. flexneri and SFN6B viability were monitored hourly for consecutive 7 hours and extended viability study for consecutive 4 days. Absorbance of all bacteria spiked water samples were taken to monitor the bacteria count. Results showed reduction in the absorbance of the SFN6B treated water sample as compared to negative control, indicating reduction in bacterial count either due to negative growth or lysis by the lytic bacteriophage. Consistent with the result, SFN6B titer increases for first two days. However, prolong incubation of these cultures reaches equilibrium, between phage and bacteria. Temperature and water sample source also influence the interaction between S. flexneri and SFN6B. Stronger interaction was observed in 37oC as compared to room temperature, where higher bacteria count and phage titer increase were recorded. Availability of nutrient in water sample also plays a crucial role in the interaction between bacteria and phage. Higher nutrient level, such as lake and river waters were observed to give better infectivity and viability of both bacteria and phage as compared to tab water. It is believed that S. flexneri continue to remain viable and able to grow in the present of SFN6B bacteriophage, but the number was closely regulated by surrounding phages. This allows better understanding of the characteristics of SFN6B that could serve as the basis for future studies and applications.

Keywords: bacteriophage, Shigella flexneri, infection, microbial diversity

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Authors: A. M. Daneshian Moghaddam, J. Shayegh, J. Dolghari Sharaf

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This study was conducted to assessment the antibacterial activities of different part of basil essential oil on the standard gram-negative bacteria include Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, and gram-positive ones including Bacillus cereus, Staphylococcus aureus, and Listeria monocytogen. The basil essential oil was provided from two part of plant (leaf and herb) at the two different developmental stage. The antibacterial properties of basil essential oil was studied Also agar disk diffusion, minimal inhibition concentration (MIC) and minimum bactericidal concentration (MBC) were detected. The results of agar disk diffusion tests showed the inhibition zones as follow: Listeria monocytogen 17.11-17.42 mm, St. aureus 29.20-30.56 mm, B. cereus 14.73-16.06 mm, E. coli 21.60-23.58 mm, Salmonella typhi 21.63-24.80 mm and for P. aeruginosa the maximum inhibition zones were seen on leaf essential oil. From the herb part of basil almost similar results were obtained: Listeria monocytogen 17.02-17.67 mm, St. aureus 29.60-30.41 mm, B. cereus 10.66-16.11 mm, E. coli 17.48-23.54 mm, Salmonella typhi 21.58-21.64 mm and for P. aeruginosa the maximum inhibition zones were seen. The MICs for gram-positive bacteria were as: B. cereus ranging 36-18 μg/mL, S. aureus 18 μg/mL, Listeria monocytogen 18-36 μg/mL and for gram-negative bacteria of E. coli, Salmonella typhi and P. aeruginosa were 18-9 μg/mL.

Keywords: basil (Ocimum basilicum) essential oil, gram-positive and gram negative bacteria, antibacterial activity, MIC, MBC

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Authors: S. M. Abd-Altwab, M. R. Noor El-Din

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This paper aims to the synthesis of new ethoxylated amide as bactericides to prevent the growth of Gram +ve and –ve bacteria of water-in-diesel fuel nanoemulsions over a long period of time as three months. To realize it, eight kinetically stable water-in-diesel fuel nanoemulsions differing in surfactant concentrations and water contents ranging from 4 to 8 and 5 to 8 wt.,wt.,% of total weight of the nanoemulsions, respectively were formed at a temperature of 20 °C. The performance of this ethoxylated amide as bactericides agents against two strains of Gram-negative bacteria, namely, Pseudomonas aeruginosa and Escherichia coli, and two strains of Gram-positive bacteria namely, Staphylococcus aureus and Bacillus subtilis, were evaluated as antimicrobial agents. The maximum and minimum antimicrobial activities were 85 and 71 % against S. aureus and E. coli, respectively, at a concentration of 5 mg/l, pH 7, and 37 °C.

Keywords: nanoemulsion, bacteriocide, diesel fuel, emulsifier

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Authors: Virve Pääkkönen, Heidi M. Cuffaro, Leo Tjäderhane

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Objectives: Odontoblasts are the outermost cell layer of dental pulp and form the dentin. Importance of bacterial products, e.g. lipoteichoic acid (LTA), a cell wall component of Gram-positive bacteria and lipopolysaccharide (LPS), a cell wall component of Gram-negative bacteria, have been indicated in the pathogenesis of pulpitis. Gram-positive bacteria are more prevalent in superficial carious lesion while the amount gram-negative is higher in the deep lesions. Objective of this study was to investigate the effect of these bacterial products on inflammatory response of pulp tissue. Interleukins (IL) were of special interest. Various ILs have been observed in the dentin-pulp complex of carious tooth in vivo. Methods: Tissue culture method was used for testing the effect of LTA and LPS on human odontoblasts. Enzymatic isolation technique was used to extract living odontoblasts for cell cultures. DNA microarray and quantitative PCR (qPCR) were used to characterize the changes in the expression profile of the tissue cultured odontoblasts. Laser microdissection was used to cut healthy and affected dentin and odontoblast layer directly under carious lesion for experiments. Cytokine array detecting 80 inflammatory cytokines was used to analyze the protein content of conditioned culture media as well as dentin and odontoblasts from the carious teeth. Results: LPS caused increased gene expression IL-1α, and -8 and decrease of IL-1β, 12 , -15 and -16 after 1h treatment, while after 24h treatment decrease of IL-8, -11 and 23 mRNAs was observed. LTA treatment caused cell death in the tissue cultured odontoblasts but in in the cell culture but not in cell culture. Cytokine array revealed at least 2-fold down-regulation of IL-1β, -10 and -12 in response to LPS treatment. Cytokine array of odontoblasts of carious teeth, as well as LPS-treated tissue-cultured odontoblasts, revealed increased protein amounts of IL-16, epidermal growth factor (EGF), angiogenin and IGFBP-1 as well as decreased amount of fractalkine. In carious dentin, increased amount of IL-1β, EGF and fractalkine was observed, as well as decreased level of GRO-1 and HGF. Conclusion: LPS caused marked changes in the expression of inflammatory cytokines in odontoblasts. Similar changes were observed in the odontoblasts cut directly under the carious lesion. These results help to shed light on the inflammatory processes happening during caries.

Keywords: inflammation, interleukin, lipoteichoic acid, odontoblasts

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Authors: Neil Jolly, Louisa Beukes, Santiago Benito-SaEz

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Kei-apple is a tree found on the African continent. Limited information exists on the effect of alcoholic and acetous fermentation on the phytochemicals. The fruit has increased L-malic, ascorbic, and phenolic acids. Juice was co-inoculated with Schizosaccharomyces pombe and Saccharomyces cerevisiae to induce alcoholic fermentation and acetous fermentation using acetic acid bacteria. Saccharomyces cerevisiae+S. pombe wines and vinegars had highest pH. Total acidity, soluble solids and L-malic acid decreased during alcoholic and acetous fermentation with highest in S. cerevisiae wines and vinegars. Volatile acidity was highest in S. pombe vinegars but not different from S. cerevisiae and S. cerevisiae+S. pombe. Gallic acid was highest in S. pombe wines and vinegars. Syringic acid was highest in S. cerevisiae wines and vinegars. S. cerevisiae+S. pombe wines were highest in caffeic, p-coumaric and protocatechuic acids. Schizosaccharomyces pombe vinegars were highest in caffeic and p-coumaric acids. Ferulic and sinapic acids were highest in S. pombe and S. cerevisiae wines, respectively. Chlorogenic acid was most abundant in both wines and vinegars. Saccharomyces cerevisiae+S. pombe and S. cerevisiae had a positive effect on most phenolic acids. Saccharomyces cerevisiae +acetic acid bacteria had an increased effect on syringic and chlorogenic acids. Schizosaccharomyces pombe+acetic acid bacteria resulted in an increase in gallic, caffeic and p-coumaric acids. Acetic acid bacteria had minimal performance with respect to volatile acidity production in comparison to commercial vinegars. Acetic acid bacteria selection should therefore be reconsidered and the decrease of certain phenolic acids during acetous fermentation needs to be investigated.

Keywords: acetic acid bacteria, liquid chromatography, phenolics, saccharomyces cerevisiae, schizosaccharomyces pombe

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Authors: Mohammad Arifin, Huda Abdullah, Norshafadzila Mohammad Naim

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The fabricated PANI-(SnO₂, ZnO)/rGO nanocomposite thin films for the E. coli bacteria sensor were investigated. The nanocomposite thin films were prepared by the sol-gel method and deposited on the glass substrate using the spin-coating technique. The internal structure and surface morphology of the thin films have been analyzed by X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), and atomic force microscopy (AFM). The optical properties of the films were investigated by UV-Vis spectroscopy, Raman spectroscopy, and Fourier transform infrared spectroscopy (FTIR). The sensitivity performance was identified by measuring the changing conductivity before and after the incubation of E. coli bacteria using current-voltage (I-V) and cyclic voltammetry (C-V) measurements.

Keywords: PANI-(SnO₂, ZnO)/rGO, nanocomposite, bacteria sensor, thin films

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Authors: Bruna Goncalves, Jennifer Henck, Romulo Uliana, Eliana Kamimura, Carlos Oliveira, Carlos Corassin

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Studies have shown evidence of human exposure to aflatoxin M1 due to the consumption of contaminated milk and dairy products (mainly cheeses). This poses a great risk to public health, since milk and milk products are frequently consumed by a portion of the population considered immunosuppressed, children and the elderly. Knowledge of the negative impacts of aflatoxins on health and economics has led to investigations of strategies to prevent their formation in food, as well as to eliminate, inactivate or reduce the bioavailability of these toxins in contaminated products This study evaluated the effect of microbiological methods using lactic acid bacteria on aflatoxin M1 (AFM1) reduction in Minas Frescal cheese (typical Brazilian product, being among the most consumed cheeses in Brazil) spiked with 1 µg/L AFM1. Inactivated lactic acid bacteria (0,5%, v/v de L. rhamnosus e L. lactis) were added during the cheese production process. Nine cheeses were produced, divided into three treatments: negative controls (without AFM1 or lactic acid bacteria), positive controls (AFM1 only), and lactic acid bacteria + AFM1. Samples of cheese were collected on days 2, 10, 20 and 30 after the date of production and submitted to composition analyses and determination of AFM1 by high-performance liquid chromatography. The reductions of AFM1 in cheese by lactic acid bacteria at the end of the trial indicate a potential application of inactivated lactic acid bacteria in reducing the bioavailability of AFM1 in Minas frescal cheese without physical-chemical and microbiological modifications during the 30-day experimental period. The authors would like to thank São Paulo Research Foundation – FAPESP (grants #2017/20081-6 and #2017/19683-1).

Keywords: aflatoxin, milk, minas frescal cheese, decontamination

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Authors: Stefano Barone

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Seismic retrofit of buildings through base isolation represents a consolidated protection strategy against earthquakes. It consists in decoupling the ground motion from that of the structure and introducing anti-seismic devices at the base of the building, characterized by high horizontal flexibility and medium/high dissipative capacity. This allows to protect structural elements and to limit damages to non-structural ones. For these reasons, full functionality is guaranteed after an earthquake event. Base isolation is applied extensively to both new and existing buildings. For the latter, it usually does not require any interruption of the structure use and occupants evacuation, a special advantage for strategic buildings such as schools, hospitals, and military buildings. This paper describes the application of seismic isolation to three existing masonry buildings in Italy: Villa “La Maddalena” in Macerata (Marche region), “Giacomo Matteotti” and “Plinio Il Giovane” school buildings in Perugia (Umbria region). The seismic hazard of the sites is characterized by a Peak Ground Acceleration (PGA) of 0.213g-0.287g for the Life Safety Limit State and between 0.271g-0.359g for the Collapse Limit State. All the buildings are isolated with a combination of free sliders type TETRON® CD with confined elastomeric disk and anti-seismic rubber isolators type ISOSISM® HDRB to reduce the eccentricity between the center of mass and stiffness, thus limiting torsional effects during a seismic event. The isolation systems are designed to lengthen the original period of vibration (i.e., without isolators) by at least three times and to guarantee medium/high levels of energy dissipation capacity (equivalent viscous damping between 12.5% and 16%). This allows the structures to resist 100% of the seismic design action. This article shows the performances of the supplied anti-seismic devices with particular attention to the experimental dynamic response. Finally, a special focus is given to the main site activities required to isolate a masonry building.

Keywords: retrofit, masonry buildings, seismic isolation, energy dissipation, anti-seismic devices

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