Search results for: bacterial inhibition

Authors: Kalaumari Mayoral-Peña, Ana I. Montejano-Montelongo, Josué Reyes-Muñoz, Gonzalo A. Ortiz-Mancilla, Mayrin Rodríguez-Cruz, Víctor Hernández-Villalobos, Jesús A. Guzmán-López, Santiago García-Jacobo, Iván Licona-Vázquez, Grisel Fierros-Romero, Rosario Flores-Vallejo

Abstract:

The group B-lactamic antibiotics include some of the most frequently used small drug molecules against bacterial infections. Nevertheless, an alarming decrease in their efficacy has been reported due to the emergence of antibiotic-resistant bacteria. Infections caused by bacteria expressing extended Spectrum B-lactamases (ESBLs) are difficult to treat and account for higher morbidity and mortality rates, delayed recovery, and high economic burden. According to the Global Report on Antimicrobial Resistance Surveillance, it is estimated that mortality due to resistant bacteria will ascend to 10 million cases per year worldwide. These facts highlight the importance of developing low-cost and readily accessible detection methods of drug-resistant ESBLs bacteria to prevent their spread and promote accurate and fast diagnosis. Bacterial detection is commonly done using molecular diagnostic techniques, where PCR stands out for its high performance. However, this technique requires specialized equipment not available everywhere, is time-consuming, and has a high cost. Loop-Mediated Isothermal Amplification (LAMP) is an alternative technique that works at a constant temperature, significantly decreasing the equipment cost. It yields double-stranded DNA of several lengths with repetitions of the target DNA sequence as a product. Although positive and negative results from LAMP can be discriminated by colorimetry, fluorescence, and turbidity, there is still a large room for improvement in the point-of-care implementation. DNA origami is a technique that allows the formation of 3D nanometric structures by folding a large single-stranded DNA (scaffold) into a determined shape with the help of short DNA sequences (staples), which hybridize with the scaffold. This research aimed to generate DNA origami structures using LAMP products as scaffolds to improve the sensitivity to detect ESBLs in point-of-care diagnosis. For this study, the coding sequence of the CTM-X-15 ESBL of E. coli was used to generate the LAMP products. The set of LAMP primers were designed using PrimerExplorerV5. As a result, a target sequence of 200 nucleotides from CTM-X-15 ESBL was obtained. Afterward, eight different DNA origami structures were designed using the target sequence in the SDCadnano and analyzed with CanDo to evaluate the stability of the 3D structures. The designs were constructed minimizing the total number of staples to reduce costs and complexity for point-of-care applications. After analyzing the DNA origami designs, two structures were selected. The first one was a zig-zag flat structure, while the second one was a wall-like shape. Given the sequence repetitions in the scaffold sequence, both were able to be assembled with only 6 different staples each one, ranging between 18 to 80 nucleotides. Simulations of both structures were performed using scaffolds of different sizes yielding stable structures in all the cases. The generation of the LAMP products were tested by colorimetry and electrophoresis. The formation of the DNA structures was analyzed using electrophoresis and colorimetry. The modeling of novel detection methods through bioinformatics tools allows reliable control and prediction of results. To our knowledge, this is the first study that uses LAMP products and DNA-origami in combination to delect ESBL-producing bacterial strains, which represent a promising methodology for diagnosis in the point-of-care.

Keywords: beta-lactamases, antibiotic resistance, DNA origami, isothermal amplification, LAMP technique, molecular diagnosis

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Authors: Razieh Niazmand, Dina Sadat Mousavian, Parvin Sharayei

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This study investigated the effect of carboxymethyl cellulose (CMC), tragacanth, and saalab hydrocolloids in two concentrations (0.3%, 0.7%) and different frying media, refined canola oil (RCO), RCO + 1% bene kernel oil (BKO), and RCO + 1 mg/l unsaponifiable matter (USM) of BKO on acrylamide formation in fried potato slices. The hydrocolloid coatings significantly reduced acrylamide formation in potatoes fried in all oils. Increasing the hydrocolloid concentration from 0.3% to 0.7% produced no effective inhibition of acrylamide. The 0.7 % CMC solution was identified as the most promising inhibitor of acrylamide formation in RCO oil, with a 62.9% reduction in acrylamide content. The addition of BKO or USM to RCO led to a noticeable reduction in the acrylamide level in fried potato slices. The findings suggest that a 0.7% CMC solution and RCO+USM are promising inhibitors of acrylamide formation in fried potato products.

Keywords: CMC, frying, potato, saalab, tracaganth

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Authors: Eduardo Lanzagorta Garcia, Chaitra Venkatesh, Romina Pezzoli, Laura Gabriela Rodriguez Barroso, Declan Devine, Margaret E. Brennan Fournet

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New antimicrobial interventions are urgently required to combat rising global health and medical infection challenges. Here, an innovative antimicrobial technology, providing price competitive alternatives to antibiotics and readily integratable with currently technological systems is presented. Two cutting edge antimicrobial materials, antimicrobial peptides (AMPs) and uncompromised sustained Ag+ action from triangular silver nanoplates (TSNPs) reservoirs, are merged for versatile effective antimicrobial action where current approaches fail. Antimicrobial peptides (AMPs) exist widely in nature and have recently been demonstrated for broad spectrum of activity against bacteria, viruses, and fungi. TSNP’s are highly discrete, homogenous and readily functionisable Ag+ nanoreseviors that have a proven amenability for operation within in a wide range of bio-based settings. In a design for advanced antimicrobial sustainable plastics, antimicrobial TSNPs are formulated for processing within biodegradable biopolymers. Histone H5 AMP was selected for its reported strong antimicrobial action and functionalized with the TSNP (AMP-TSNP) in a similar fashion to previously reported TSNP biofunctionalisation methods. A synergy between the propensity of biopolymers for degradation and Ag+ release combined with AMP activity provides a novel mechanism for the sustained antimicrobial action of biopolymeric thin films. Nanoplates are transferred from aqueous phase to an organic solvent in order to facilitate integration within hydrophobic polymers. Extrusion is used in combination with calendering rolls to create thin polymerc film where the nanoplates are embedded onto the surface. The resultant antibacterial functional films are suitable to be adapted for food packing and biomedical applications. TSNP synthesis were synthesized by adapting a previously reported seed mediated approach. TSNP synthesis was scaled up for litre scale batch production and subsequently concentrated to 43 ppm using thermally controlled H2O removal. Nanoplates were transferred from aqueous phase to an organic solvent in order to facilitate integration within hydrophobic polymers. This was acomplised by functionalizing the TSNP with thiol terminated polyethylene glycol and using centrifugal force to transfer them to chloroform. Polycaprolactone (PCL) and Polylactic acid (PLA) were individually processed through extrusion, TSNP and AMP-TSNP solutions were sprayed onto the polymer immediately after exiting the dye. Calendering rolls were used to disperse and incorporate TSNP and TSNP-AMP onto the surface of the extruded films. Observation of the characteristic blue colour confirms the integrity of the TSNP within the films. Antimicrobial tests were performed by incubating Gram + and Gram – strains with treated and non-treated films, to evaluate if bacterial growth was reduced due to the presence of the TSNP. The resulting films successfully incorporated TSNP and AMP-TSNP. Reduced bacterial growth was observed for both Gram + and Gram – strains for both TSNP and AMP-TSNP compared with untreated films indicating antimicrobial action. The largest growth reduction was observed for AMP-TSNP treated films demonstrating the additional antimicrobial activity due to the presence of the AMPs. The potential of this technology to impede bacterial activity in food industry and medical surfaces will forge new confidence in the battle against antibiotic resistant bacteria, serving to greatly inhibit infections and facilitate patient recovery.

Keywords: antimicrobial, biodegradable, peptide, polymer, nanoparticle

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Authors: Palwinder Singh

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Beyond the conventional mode of working with anti-inflammatory agents through enzyme inhibition, herein, an alternate substrate of cyclooxygenase-2 was developed. Proline centered pentapeptide iso-conformational to arachidonic acid exhibited appreciable selectivity for COX-2 overcoming acetic acid and formalin induced pain in rats to almost 80% and was treated as a substrate by the enzyme. Remarkably, COX-2 metabolized the pentapeptide into small fragments consisting mainly of di- and tri-peptides that ensured the safe breakdown of the peptide under in-vivo conditions. The kinetic parameter Kcat/Km for COX-2 mediated metabolism of peptide 6.3 x 105 M-1 s-1 was quite similar to 9.5 x 105 M-1 s-1 for arachidonic acid. Evidenced by the dynamic molecular studies and the use of Y385F COX-2, it was observed that the breakage of the pentapeptide has probably taken place through H-bond activation of the peptide bond by the side chains of Y385 and S530.

Keywords: small peptides, anti-inflammatory agents, cyclooxygenase-2, unnatural substrates

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Authors: Nuha Mansour Alhazmi

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Replacement of petro-based plastics by a biodegradable plastic are vastly growing process. Poly-β-hydroxybutyrate (PHB) is a biodegradable biopolymer, synthesized by some bacterial genera. The objective of the current study is to explore the ability of some fungi to biodegrade PHB. The degradation of (PHB) was detected in Petri dish by the formation of a clear zone around the fungal colonies due to the production of depolymerase enzyme which has an interesting role in the PHB degradation process. Among 10 tested fungi, the most active PHB biodegraded fungi were identified as Trichoderma asperellum using morphological and molecular characters. The highest PHB degradation was at 25°C, pH 7.5 after 7 days of incubation for the tested fungi. Finally, the depolymerase enzyme was isolated, purified using column chromatography and characterized. In conclusion, PHB can be biodegraded in solid and liquid medium using depolymerase enzyme from T. asperellum.

Keywords: degradation, depolymerase enzyme, PHB, Trichoderma asperellum

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Authors: Akrpoum Souad, Lalaoui Korrichi

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Vicia faba L.,Vaccinium macrocarpon, Punica granatum, Lavandula officinalis, Artemisia absinthium, Linum capitatum and Camellia sinensis were frequently used in our alimentation. In this study, we have tested the antimicrobial activity of their ethanolic and methanolic extracts on some pathogen bacteria, then their ability to in vivo inhibit the growth of Strepcoccus pneumonia. The phytochemical screening has given the composition of the most active extracts. According to the obtained results, the ethanolic extract of Lavendula. officinalis and A absinthium has shown an inhibition of all the tested strains of becteria3. The ethanolic extract of L. officinalis has given the highest activity against S. pneumoniae, followed by the methanolic extract of C. sinensis 1, 2 and P. granatum. The phytochemical screening showed that the most active extracts contained mainly naturels compounds.

Keywords: plants, extracts, antimicrobial activity, streptococcus pneumoniae, phytochemical screening

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Authors: M. Ouassaf, S. Belaidi

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Aromatase is an estrogen biosynthetic enzyme belonging to the cytochrome P450 family, which catalyzes the limiting step in the conversion of androgens to estrogens. As it is relevant for the promotion of tumor cell growth. A set of thirty 1,2,3-triazole derivatives was used in the quantitative structure activity relationship (QSAR) study using regression multiple linear (MLR), We divided the data into two training and testing groups. The results showed a good predictive ability of the MLR model, the models were statistically robust internally (R² = 0.982) and the predictability of the model was tested by several parameters. including external criteria (R²pred = 0.851, CCC = 0.946). The knowledge gained in this study should provide relevant information that contributes to the origins of aromatase inhibitory activity and, therefore, facilitates our ongoing quest for aromatase inhibitors with robust properties.

Keywords: aromatase inhibitors, QSAR, MLR, 1, 2, 3-triazole

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Authors: Christopher Latella, Ashlee M. Hendy, Dan Vander Westhuizen, Wei-Peng Teo

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Introduction: Neural adaptations following resistance training interventions have been widely investigated, however the evidence regarding the mechanisms of early adaptation are less clear. Understanding neural responses from an acute resistance training session is pivotal in the prescription of frequency, intensity and volume in applied strength and conditioning practice. Therefore the primary aim of this study was to investigate the time course of neurophysiological mechanisms post training against current super compensation theory, and secondly, to examine whether these responses reflect neural adaptations observed with resistance training interventions. Methods: Participants (N=14) completed a randomised, counterbalanced crossover study comparing; control, strength and hypertrophy conditions. The strength condition involved 3 x 5RM leg extensions with 3min recovery, while the hypertrophy condition involved 3 x 12 RM with 60s recovery. Transcranial magnetic stimulation (TMS) and peripheral nerve stimulation were used to measure excitability of the central and peripheral neural pathways, and maximal voluntary contraction (MVC) to quantify strength changes. Measures were taken pre, immediately post, 10, 20 and 30 mins and 1, 2, 6, 24, 48, 72 and 96 hrs following training. Results: Significant decreases were observed at post, 10, 20, 30 min, 1 and 2 hrs for both training groups compared to control group for force, (p <.05), maximal compound wave; (p < .005), silent period; (p < .05). A significant increase in corticospinal excitability; (p < .005) was observed for both groups. Corticospinal excitability between strength and hypertrophy groups was near significance, with a large effect (η2= .202). All measures returned to baseline within 6 hrs post training. Discussion: Neurophysiological mechanisms appear to be significantly altered in the period 2 hrs post training, returning to homeostasis by 6 hrs. The evidence suggests that the time course of neural recovery post resistance training occurs 18-40 hours shorter than previous super compensation models. Strength and hypertrophy protocols showed similar response profiles with current findings suggesting greater post training corticospinal drive from hypertrophy training, despite previous evidence that strength training requires greater neural input. The increase in corticospinal drive and decrease inl inhibition appear to be a compensatory mechanism for decreases in peripheral nerve excitability and maximal voluntary force output. The changes in corticospinal excitability and inhibition are akin to adaptive processes observed with training interventions of 4 wks or longer. It appears that the 2 hr recovery period post training is the most influential for priming further neural adaptations with resistance training. Secondly, the frequency of prescribed resistance sessions can be scheduled closer than previous super compensation theory for optimal strength gains.

Keywords: neural responses, resistance training, super compensation, transcranial magnetic stimulation

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Authors: Bibha Kumari, Nigel P. Brunton, Dilip K. Rai, Brijesh K. Tiwari

Abstract:

Edible mushrooms possess numerous functional components like homo- and hetero- β-glucans [β(1→3), β(1→4) and β(1→6) glucosidic linkages], chitins, ergosterols, bioactive polysaccharides and peptides imparting health beneficial properties to mushrooms. Some of the proven biological activities of mushroom extracts are antioxidant, antimicrobial, immunomodulatory, cholesterol lowering activity by inhibiting a key cholesterol metabolism enzyme i.e. 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGCR), angiotensin I-converting enzyme (ACE) inhibition. Application of novel extraction technologies like pulsed electric field (PEF) and high power ultrasound offers clean, green, faster and efficient extraction alternatives with enhanced and good quality extracts. Sequential PEF followed by ultrasound assisted extraction (UAE) were applied to recover bioactive enriched fractions from industrial white button mushroom (Agaricus bisporus) stalk waste using environmentally friendly and GRAS solvents i.e. water and water/ethanol combinations. The PEF treatment was carried out at 60% output voltage, 2 Hz frequency for 500 pulses of 20 microseconds pulse width, using KCl salt solution of 0.6 mS/cm conductivity by the placing 35g of chopped fresh mushroom stalks and 25g of salt solution in the 4x4x4cm3 treatment chamber. Sequential UAE was carried out on the PEF pre-treated samples using ultrasonic-water-bath (USB) of three frequencies (25 KHz, 35 KHz and 45 KHz) for various treatment times (15-120 min) at 80°C. Individual treatment using either PEF or UAE were also investigation to compare the effect of each treatment along with the combined effect on the recovery and bioactivity of the crude extracts. The freeze dried mushroom stalk powder was characterised for proximate compositional parameters (dry weight basis) showing 64.11% total carbohydrate, 19.12% total protein, 7.21% total fat, 31.2% total dietary fiber, 7.9% chitin (as glucosamine equivalent) and 1.02% β-glucan content. The total phenolic contents (TPC) were determined by the Folin-Ciocalteu procedure and expressed as gallic-acid-equivalents (GAE). The antioxidant properties were ascertained using DPPH and FRAP assays and expressed as trolox-equivalents (TE). HMGCR activity and molecular mass of β-glucans will be measured using the commercial HMG-CoA Reductase Assay kit (Sigma-Aldrich) and size exclusion chromatography (HPLC-SEC), respectively. Effects of PEF, UAE and their combination on the antioxidant capacity, HMGCR inhibition and β-glucans content will be presented.

Keywords: β-glucan, mushroom stalks, pulsed electric field (PEF), ultrasound assisted extraction (UAE)

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Authors: V. K. Rajaletchumy, S. L. Chia, M. I. Setyawati, M. S. Muthu, S. S. Feng, D. T. Leong

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Triple negative breast cancers (TNBC) can be classified as one of the most aggressive with a high rate of local recurrences and systematic metastases. TNBCs are insensitive to existing hormonal therapy or targeted therapies such as the use of monoclonal antibodies, due to the lack of oestrogen receptor (ER) and progesterone receptor (PR) and the absence of overexpression of human epidermal growth factor receptor 2 (HER2) compared with other types of breast cancers. The absence of targeted therapies for selective delivery of therapeutic agents into tumours, led to the search for druggable targets in TNBC. In this study, we developed a targeted micellar system of cetuximab-conjugated micelles of D-α-tocopheryl polyethylene glycol succinate (vitamin E TPGS) for targeted delivery of docetaxel as a model anticancer drug for the treatment of TNBCs. We examined the efficacy of our micellar system in xenograft models of triple negative breast cancers and explored the effect of the micelles on post-treatment tumours in order to elucidate the mechanism underlying the nanomedicine treatment in oncology. The targeting micelles were found preferentially accumulated in tumours immediately after the administration of the micelles compare to normal tissue. The fluorescence signal gradually increased up to 12 h at the tumour site and sustained for up to 24 h, reflecting the increases in targeted micelles (TPFC) micelles in MDA-MB-231/Luc cells. In comparison, for the non-targeting micelles (TPF), the fluorescence signal was evenly distributed all over the body of the mice. Only a slight increase in fluorescence at the chest area was observed after 24 h post-injection, reflecting the moderate uptake of micelles by the tumour. The successful delivery of docetaxel into tumour by the targeted micelles (TPDC) exhibited a greater degree of tumour growth inhibition than Taxotere® after 15 days of treatment. The ex vivo study has demonstrated that tumours treated with targeting micelles exhibit enhanced cell cycle arrest and attenuated proliferation compared with the control and with those treated non-targeting micelles. Furthermore, the ex vivo investigation revealed that both the targeting and non-targeting micellar formulations shows significant inhibition of cell migration with migration indices reduced by 0.098- and 0.28-fold, respectively, relative to the control. Overall, both the in vivo and ex vivo data increased the confidence that our micellar formulations effectively targeted and inhibited EGF-overexpressing MDA-MB-231 tumours.

Keywords: biodegradable polymers, cancer nanotechnology, drug targeting, molecular biomaterials, nanomedicine

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Authors: C. C. Castro, T. Senechal, D. Lahem, A. L. Hantson

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Formaldehyde is one of the most representative volatile organic compounds present in the indoor air of residential units and workplaces. Increased attention has been given to this toxic compound because of its carcinogenic effect in health. Biological or enzymatic transformation is being explored to degrade this pollutant. Pseudomonas putida is a bacteria able to synthesize formaldehyde dehydrogenase, an enzyme known to use formaldehyde as a substrate and transform it into less toxic compounds. The immobilization of bacterial cells in the surface of different supports through spraying or dip-coating is herein proposed. The determination of the enzymatic activity on the coated surfaces was performed as well as the study of its effect on formaldehyde degradation in an isolated chamber. Results show that the incorporation of microbial cells able to synthesize depolluting enzymes can be an innovative, low-cost, effective and environmentally friendly solution for indoor air depollution.

Keywords: cells encapsulation, formaldehyde, formaldehyde dehydrogenase, indoor air depollution

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Authors: Sonu Gandhi, Neena Capalash, Prince Sharma, C. Raman Suri

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A sensitive chemiluminescence immunoassay (CIA) for heroin and its major metabolites is reported. The method is based on the competitive reaction of horseradish peroxidase (HRP)-labeled anti-MAM antibody and free drug in spiked urine samples. A hapten-protein conjugate was synthesized by using acidic derivative of monoacetyl morphine (MAM) coupled to carrier protein BSA and was used as an immunogen for the generation of anti-MAM (monoacetyl morphine) antibody. A high titer of antibody (1:64,0000) was obtained and the relative affinity constant (Kaff) of antibody was 3.1×107 l/mol. Under the optimal conditions, linear range and reactivity for heroin, mono acetyl morphine (MAM), morphine and codeine were 0.08, 0.09, 0.095 and 0.092 ng/mL respectively. The developed chemiluminescence inhibition assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml.

Keywords: heroin, metabolites, chemiluminescence immunoassay, horse radish peroxidase

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Authors: Tanmaya Pradhan, K. Midhun, M. Joy Thomas

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Increasing the shelf life and improving the quality are important objectives for the success of packaged liquid food industry. One of the methods by which this can be achieved is by deactivating the micro-organisms present in the liquid food through pasteurization. Pasteurization is done by heating, but some serious disadvantages such as the reduction in food quality, flavour, taste, colour, etc. were observed because of heat treatment, which leads to the development of newer methods instead of pasteurization such as treatment using UV radiation, high pressure, nuclear irradiation, pulsed electric field, etc. In recent years the use of the pulsed electric field (PEF) for inactivation of the microbial content in the food is gaining popularity. PEF uses a very high electric field for a short time for the inactivation of microorganisms, for which we require a high voltage pulsed power source. Pulsed power sources used for PEF treatments are usually in the range of 5kV to 50kV. Different pulse shapes are used, such as exponentially decaying and square wave pulses. Exponentially decaying pulses are generated by high power switches with only turn-on capacity and, therefore, discharge the total energy stored in the capacitor bank. These pulses have a sudden onset and, therefore, a high rate of rising but have a very slow decay, which yields extra heat, which is ineffective in microbial inactivation. Square pulses can be produced by an incomplete discharge of a capacitor with the help of a switch having both on/off control or by using a pulse forming network. In this work, a pulsed power-based system is designed with the help of high voltage capacitors and solid-state switches (IGBT) for the inactivation of pathogenic micro-organism in liquid food such as fruit juices. The high voltage generator is based on the Marx generator topology, which can produce variable amplitude, frequency, and pulse width according to the requirements. Liquid food is treated in a chamber where pulsed electric field is produced between stainless steel electrodes using the pulsed output voltage of the supply. Preliminary bacterial inactivation tests were performed by subjecting orange juice inoculated with Escherichia Coli bacteria. With the help of the developed pulsed power source and the chamber, the inoculated orange has been PEF treated. The voltage was varied to get a peak electric field up to 15kV/cm. For a total treatment time of 200µs, a 30% reduction in the bacterial count has been observed. The detailed results and analysis will be presented in the final paper.

Keywords: Escherichia coli bacteria, high voltage generator, microbial inactivation, pulsed electric field, pulsed forming line, solid-state switch

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Authors: Lina Wu

Abstract:

The excessive discharge of nitrogen in sewage greatly intensifies the eutrophication of water bodies and threatens water quality. Nitrogen pollution control has become a global concern. The concentration of nitrogen in water is reduced by converting ammonia nitrogen, nitrate nitrogen and nitrite nitrogen into nitrogen-containing gas through biological treatment, physicochemical treatment and oxidation technology. However, some wastewater containing high ammonia nitrogen including landfill leachate, is difficult to be treated by traditional nitrification and denitrification because of its high COD content. The core process of denitrification is that denitrifying bacteria convert nitrous acid produced by nitrification into nitrite under anaerobic conditions. Still, its low-carbon nitrogen does not meet the conditions for denitrification. Many studies have shown that the natural autotrophic anammox bacteria can combine nitrous and ammonia nitrogen without a carbon source through functional genes to achieve total nitrogen removal, which is very suitable for removing nitrogen from leachate. In addition, the process also saves a lot of aeration energy consumption than the traditional nitrogen removal process. Therefore, anammox plays an important role in nitrogen conversion and energy saving. The short-range nitrification and denitrification coupled with anaerobic ammoX ensures total nitrogen removal. It improves the removal efficiency, meeting the needs of society for an ecologically friendly and cost-effective nutrient removal treatment technology. In recent years, research has found that the symbiotic system has more water treatment advantages because this process not only helps to improve the efficiency of wastewater treatment but also allows carbon dioxide reduction and resource recovery. Microalgae use carbon dioxide dissolved in water or released through bacterial respiration to produce oxygen for bacteria through photosynthesis under light, and bacteria, in turn, provide metabolites and inorganic carbon sources for the growth of microalgae, which may lead the algal bacteria symbiotic system save most or all of the aeration energy consumption. It has become a trend to make microalgae and light-avoiding anammox bacteria play synergistic roles by adjusting the light-to-dark ratio. Microalgae in the outer layer of light particles block most of the light and provide cofactors and amino acids to promote nitrogen removal. In particular, myxoccota MYX1 can degrade extracellular proteins produced by microalgae, providing amino acids for the entire bacterial community, which helps anammox bacteria save metabolic energy and adapt to light. As a result, initiating and maintaining the process of combining dominant algae and anaerobic denitrifying bacterial communities has great potential in treating landfill leachate. Chlorella has a brilliant removal effect and can withstand extreme environments in terms of high ammonia nitrogen, high salt and low temperature. It is urgent to study whether the algal mud mixture rich in denitrifying bacteria and chlorella can greatly improve the efficiency of landfill leachate treatment under an anaerobic environment where photosynthesis is stopped. The optimal dilution concentration of simulated landfill leachate can be found by determining the treatment effect of the same batch of bacteria and algae mixtures under different initial ammonia nitrogen concentrations and making a comparison. High-throughput sequencing technology was used to analyze the changes in microbial diversity, related functional genera and functional genes under optimal conditions, providing a theoretical and practical basis for the engineering application of novel bacteria-algae symbiosis system in biogas slurry treatment and resource utilization.

Keywords: nutrient removal and recovery, leachate, anammox, Partial nitrification, Algae-bacteria interaction

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Authors: M. Djekoun, H. Berrebah, M. R. Djebar

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Fusarium wilt is destructive disease of cereal crops with small grains. It affects yields but also the quality of the crop and economic losses arising are often very heavy. Chemical control is currently one of the most effective ways to fight against these diseases. In this study, the efficacy of three fungicides (tebuconazole, thiram, and fludioxonil-difenoconazole mixture) was tested. In vitro, on the phytopathogenic Fusarium spp. isolated from seeds of wheat. The active ingredients were tested at different concentrations: 0.06, 1.39, 2.79, 5.58, and 11.16 mg/l for tebuconazole, 0.035, 0.052, 0.105, 0.21, and 0.42 mg/l for thiram and finally, for the mixture fludioxonil-difenoconazole 4 concentrations were tested: 0.05, 0.1, 0.5 and 1 mg/l. Toxicity responses were expressed as effective concentration, which inhibits mycelial growth by 50%, (EC50). Of the three selected fungicides, thirame proved to be the most effective with EC50 value of the order of 0,15 mg/l followed by the mixture of fludioxonil-difenoconazole with 0,27mg/l and finally tebuconazole with a value of 3.79 mg/l.

Keywords: Fusarium spp., thiram, tebuconazole, fludioxonil, difenoconazole, percentage of inhibition, EC50

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Authors: Michelle Belinda S. Weerawardana, Gobika Thiripuranathar, Priyani A. Paranagama

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Most of fresh vegetables and fruits available in the retail markets undergo a physiological disorder in its appearance and coloration, which indeed discourages consumer purchase. A loss of millions of dollars yearly to the food industry had been due to this pronounced color reaction called Enzymatic Browning which is driven due to the catalytic activity by an oxidoreductase enzyme, polyphenol oxidase (PPO). The enzyme oxidizes the phenolic compounds which are abundantly available in fruits and vegetables as substrates into quinones, which could react with proteins in its surrounding to generate black pigments, called melanins, which are highly UV-active compounds. Annona muricata (Katu anoda) and Musa acuminata (Ash plantains) is a fruit and a vegetable consumed by Sri Lankans widely due to their high nutritional values, medicinal properties and economical importance. The objective of the present study was to evaluate and determine the effective natural anti-browning inhibitors that could prevent PPO activity in the selected fruit and vegetable. Enzyme extracts from Annona muricata (Katu anoda) and Musa acuminata (Ash plantains), were prepared by homogenizing with analytical grade acetone, and pH of each enzyme extract was maintained at 7.0 using a phosphate buffer. The extracts of inhibitors were prepared using powdered ginger rhizomes and essential oil from the bark of Cinnamomum zeylanicum. Water extracts of ginger were prepared and the essential oil from Ceylon cinnamon bark was extracted using steam distillation method. Since the essential oil is not soluble in water, 0.1µl of cinnamon bark oil was mixed with 0.1µl of Triton X-100 emulsifier and 5.00 ml of water. The effect of each inhibitor on the PPO activity was investigated using catechol (0.1 mol dm-3) as the substrate and two samples of enzyme extracts prepared. The dosages of the prepared Cinnamon bark oil, and ginger (2 samples) which were used to measure the activity were 0.0035 g/ml, 0.091 g/ml and 0.087 g/ml respectively. The measurements of the inhibitory activity were obtained at a wavelength of 525 nm using the UV-visible spectrophotometer. The results evaluated thus revealed that % inhibition observed with cinnamon bark oil, and ginger for Annona muricata was 51.97%, and 60.90% respectively. The effects of cinnamon bark oil, and ginger extract on PPO activity of Musa acuminata were 49.51%, and 48.10%. The experimental findings thus revealed that Cinnamomum zeylanicum bark oil was a more effective inhibitor for PPO enzyme present in Musa acuminata and ginger was effective for PPO enzyme present in Annona muricata. Overall both the inhibitors were proven to be more effective towards the activities of PPO enzyme present in both samples. These inhibitors can thus be corroborated as effective, natural, non-toxic, anti-browning extracts, which when added to the above fruit and vegetable will increase the shelf life and also the acceptance of the product by the consumers.

Keywords: anti-browning agent, enzymatic browning, inhibitory activity, polyphenol oxidase

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Authors: Leila Fozouni, Anahita Mazandarani

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Background: Maternal infections are the most common cause of infections in infants, and the level of infection and its severity highly depends on the degree of colonization of the bacteria in the mother; so, the occurrence of aggressive diseases is not unpredictable in mothers with very high colonization. Group B Streptococcus is part of the normal flora of the gastrointestinal and genital tracts in women and is the leading cause of septicemia and meningitis in newborns. Today Zinc oxide nanoparticle is regarded as one of the most commonly used and safest nanoparticles for defeating Gram-positive and Gram-negative bacteria. This study aims to determine the antibacterial effects of Zinc oxide on the growth of drug-resistant group B Streptococcus strains isolated from pregnant women. Materials and Methods: This cross-sectional study was conducted on 150 pregnant women of 28–37 weeks admitted to seven hospitals and maternity wards in Golestan province, northeast of Iran. For bacterial identification, rectovaginal swabs were firstly inoculated to the Todd-Hewitt Broth and cultured in blood agar (containing 5% sheep blood). Then microbiologic and PCR methods were performed to detect group B Streptococci. Disk diffusion and broth microdilution tests were used to determine the bacterial susceptibility to antibiotics according to CLSI M100(2021) criteria. The antibacterial properties of Zinc oxide nanoparticles were evaluated using the agar well-diffusion method. Results: The prevalence of group B Streptococcus was 18% in pregnant women. Out of twenty-seven positive cultures, 62.96% were higher than thirty years old. Ninety percent and 45% of isolates were resistant to clindamycin and erythromycin, respectively, and susceptibility to cefazolin was 71%. In addition, susceptibility to ampicillin and penicillin were 74% and 55%, respectively. The results showed that 82% of erythromycin-resistant, 92% clindamycin-resistant, and 78% of cefazolin-resistant isolates were eliminated by zinc oxide nanoparticles at a concentration of 100 mg/L of the nanoparticle. Furthermore, ZnONPs could inhibit all drug-resistant isolates at a concentration of 200 mg/mL (MIC90 ≥ 200). Conclusion: Since the drug resistance of group B streptococci against various antibiotics is increasing, determining and investigating the drug-resistance pattern of this bacterium to different antibiotics in order to prevent arbitrary consumption of antibiotics by pregnant women and ultimately prevent Infant mortality seems necessary. Generally, ZnONPs showed a high antimicrobial effect, and it was revealed that the bactericide effect increases upon the increase in the concentration of the nanoparticle.

Keywords: group B beta-hemolytic streptococcus, pregnant women, zinc oxide nanoparticles, drug resistance

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Authors: Indra Gonzalez Ojeda, Tatiana Quinones, Manuel Rosario, Igor Lednev, Juan Lopez Garriga

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Amyloid fibrils are stable aggregates of misfolded protein associated with many neurodegenerative disorders. It has been shown that hydrogen sulfide (H2S), inhibits the fibrillation of lysozyme through the formation of trisulfide (S-S-S) bonds. However, the overall mechanism remains elusive. Here, the concentration dependence of H2S effect was investigated using Atomic force microscopy (AFM), non-resonance Raman spectroscopy, Deep-UV Raman spectroscopy and circular dichroism (CD). It was found that small spherical aggregates with trisulfide bonds and a unique secondary structure were formed instead of amyloid fibrils when adding concentrations of 25 mM and 50 mM of H2S. This could indicate that H2S might serve as a protecting agent for the protein. However, further characterization of these aggregates and their trisulfide bonds is needed to fully unravel the function H2S has on protein fibrillation.

Keywords: amyloid fibrils, hydrogen sulfide, protein folding, raman spectroscopy

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Authors: Marcin Debowski, Marcin Zielinski, Magdalena Zielinska, Paulina Rusanowska

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The problem with digestate management is one of the most important factors influencing on the development and operation of biogas plant. Turbidity and bacterial contamination negatively affect the growth of algae, which can limit the use of the effluent in the production of algae biomass on a large scale. These problems can be overcome by cultivating of algae species resistant to environmental factors, such as Chlorella sp., Scenedesmus sp., or reducing load of organic compounds to prevent bacterial contamination. The effluent requires dilution and/or purification. One of the methods of effluent treatment is the use of a membrane technology such as microfiltration (MF), ultrafiltration (UF), nanofiltration (NF) and reverse osmosis (RO), depending on the membrane pore size and the cut off point. Membranes are a physical barrier to solids and particles larger than the size of the pores. MF membranes have the largest pores and are used to remove turbidity, suspensions, bacteria and some viruses. UF membranes remove also color, odor and organic compounds with high molecular weight. In treatment of wastewater or other waste streams, MF and UF can provide a sufficient degree of purification. NF membranes are used to remove natural organic matter from waters, water disinfection products and sulfates. RO membranes are applied to remove monovalent ions such as Na⁺ or K⁺. The effluent was used in UF for medium to cultivation of two microalgae: Chlorella sp. and Phaeodactylum tricornutum. Growth rates of Chlorella sp. and P. tricornutum were similar: 0.216 d⁻¹ and 0.200 d⁻¹ (Chlorella sp.); 0.128 d⁻¹ and 0.126 d⁻¹ (P. tricornutum), on synthetic medium and permeate from UF, respectively. The final biomass composition was also similar, regardless of the medium. Removal of nitrogen was 92% and 71% by Chlorella sp. and P. tricornutum, respectively. The fermentation effluents after UF and dilution were also used for cultivation of algae Scenedesmus sp. that is resistant to environmental conditions. The authors recommended the development of biorafinery based on the production of algae for the biogas production. There are examples of using a multi-stage membrane system to purify the liquid fraction from digestate. After the initial UF, RO is used to remove ammonium nitrogen and COD. To obtain a permeate with a concentration of ammonium nitrogen allowing to discharge it into the environment, it was necessary to apply three-stage RO. The composition of the permeate after two-stage RO was: COD 50–60 mg/dm³, dry solids 0 mg/dm³, ammonium nitrogen 300–320 mg/dm³, total nitrogen 320–340 mg/dm³, total phosphorus 53 mg/dm³. However compostion of permeate after three-stage RO was: COD < 5 mg/dm³, dry solids 0 mg/dm³, ammonium nitrogen 0 mg/dm³, total nitrogen 3.5 mg/dm³, total phosphorus < 0,05 mg/dm³. Last stage of RO might be replaced by ion exchange process. The negative aspect of membrane filtration systems is the fact that the permeate is about 50% of the introduced volume, the remainder is the retentate. The management of a retentate might involve recirculation to a biogas plant.

Keywords: digestate, membrane filtration, microalgae cultivation, Chlorella sp.

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Authors: Laura Gonzalez, Santiago Benavides, Martha Elena Londono, Ana Elisa Casas, Adriana Restrepo-Osorio

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Cotton products are sensitive to microorganisms due to its ability to retain moisture, which might cause change into the coloration, mechanical properties reduction or foul odor generation; consequently, this represents risks to the health of users. Nowadays, have been carried out researches to give antibacterial properties to textiles using different strategies, which included the use of silver nanoparticles (AgNPs). The antibacterial behavior can be affected by laundering process reducing its effectiveness. In the other way, the environmental impact generated for the synthetic antibacterial agents has motivated to seek new and more ecological ways for produce AgNPs. The aims of this work are to determine the antibacterial activity of cotton fabric functionalized with green (G) and commercial (C) AgNPs after twenty washing cycles, also to evaluate morphological and color changes. A plain weave cotton fabric suitable for dyeing and two AgNPs solutions were use. C a commercial product and G produced using an ecological method, both solutions with 0.5 mM concentration were impregnated on cotton fabric without stabilizer, at a liquor to fabric ratio of 1:20 in constant agitation during 30min and then dried at 70 °C by 10 min. After that the samples were subjected to twenty washing cycles using phosphate-free detergent simulated on agitated flask at 150 rpm, then were centrifuged and dried on a tumble. The samples were characterized using Kirby-Bauer test determine antibacterial activity against E. coli y S. aureus microorganisms, the results were registered by photographs establishing the inhibition halo before and after the washing cycles, the tests were conducted in triplicate. Scanning electron microscope (SEM) was used to observe the morphologies of cotton fabric and treated samples. The color changes of cotton fabrics in relation to the untreated samples were obtained by spectrophotometer analysis. The images, reveals the presence of inhibition halo in the samples treated with C and G AgNPs solutions, even after twenty washing cycles, which indicated a good antibacterial activity and washing durability, with a tendency to better results against to S. aureus bacteria. The presence of AgNPs on the surface of cotton fiber and morphological changes were observed through SEM, after and before washing cycles. The own color of the cotton fiber has been significantly altered with both antibacterial solutions. According to the colorimetric results, the samples treated with C lead to yellowing while the samples modified with G to red yellowing Cotton fabrics treated AgNPs C and G from 0.5 mM solutions exhibited excellent antimicrobial activity against E. coli and S. aureus with good laundering durability effects. The surface of the cotton fibers was modified with the presence of AgNPs C and G due to the presence of NPs and its agglomerates. There are significant changes in the natural color of cotton fabric due to deposition of AgNPs C and G which were maintained after laundering process.

Keywords: antibacterial property, cotton fabric, fastness to wash, Kirby-Bauer test, silver nanoparticles

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Authors: C. S. Paglia, E. Pesenti, A. Krattiger

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Commercially available methacrylate and acrylamide-based acrylic resins for injection in concrete systems have been tested with respect to the sealing performance and the rebar corrosion. Among the different resins, a methacrylate-based type of acrylic resin significantly inhibited the rebar corrosion. This was mainly caused by the relatively high pH of the resin and the resin aqueous solution. This resin also exhibited a relatively high sealing performance, in particular after exposing the resin to durability tests. The corrosion inhibition behaviour and the sealing properties after the exposition to durability tests were maintained up to one year. The other resins either promoted the corrosion of the rebar and/or exhibited relatively low sealing properties.

Keywords: acrylic resin, sealing performance, rebar corrosion, materials

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Authors: B. T. Naveen Kumar, Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, A. H. Shanthanagouda, Orawan Boodde, K. M. Shankar, Prakash Patil, Shubhkaramjeet Kaur

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Early mortality syndrome (EMS)/Acute Hepatopancreatic Necrosis Disease (AHPND) has emerged as a major obstacle for the shrimp farming around the world. It is caused by a strain of Vibrio parahaemolyticus. The possible preventive and control measure is, early and rapid detection of the pathogen in the broodstock, post-larvae and monitoring the shrimp during the culture period. Polymerase chain reaction (PCR) based early detection methods are good, but they are costly, time taking and requires a sophisticated laboratory. The present study was conducted to develop a simple, sensitive and rapid diagnostic farmer level kit for the reliable detection of AHPND in shrimp. A panel of monoclonal antibodies (MAbs) were raised against the recombinant Pir B protein (rPirB). First, an immunodot was developed by using MAbs G3B8 and Mab G3H2 which showed specific reactivity to purified r-PirB protein with no cross-reactivity to other shrimp bacterial pathogens (AHPND free Vibrio parahaemolyticus (Indian strains), V. anguillarum, WSSV, Aeromonas hydrophila, and Aphanomyces invadans). Immunodot developed using Mab G3B8 is more sensitive than that with the Mab G3H2. However, immunodot takes almost 2.5 hours to complete with several hands-on steps. Therefore, the flow-through assay (FTA) was developed by using a plastic cassette containing the nitrocellulose membrane with absorbing pads below. The sample was dotted in the test zone on the nitrocellulose membrane followed by continuos addition of five solutions in the order of i) blocking buffer (BSA) ii) primary antibody (MAb) iii) washing Solution iv) secondary antibody and v) chromogen substrate (TMB) clear purple dots against a white background were considered as positive reactions. The FTA developed using MAbG3B8 is more sensitive than that with MAb G3H2. In FTA the two MAbs showed specific reactivity to purified r-PirB protein and not to other shrimp bacterial pathogens. The FTA is simple to farmer/field level, sensitive and rapid requiring only 8-10 min for completion. Tests can be developed to kits, which will be ideal for use in biosecurity, for the first line of screening (at the port or pond site) and during monitoring and surveillance programmes overall for the good management practices to reduce the risk of the disease.

Keywords: acute hepatopancreatic necrosis disease, AHPND, flow-through assay, FTA, farmer level, immunodot, pond site, shrimp

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Authors: Harche Rima, Laoufi Nadia Aicha

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The present work consists of studying the behavior of steel, copper, and aluminum vis-à-vis the corrosion in an aqueous medium in the presence of the antifreeze COOLELF MDX -26°C. For this, we have studied the influence of the temperature and the different concentrations of the antifreeze on the corrosion of these three metals, this will last for two months by the polarization method and weight loss. In the end, we investigated the samples with the optic microscope to know their surface state. The aim of this work is the protection of contraptions. The use of antifreeze in ordinary water has a high efficiency against steel corrosion, as demonstrated by electrochemical tests (potential monitoring as a function of time and tracing polarization curves). The inhibition rate is greater than 99% for different volume concentrations, ranging from 40% to 60%. The speeds are in turn low in the order of 10-4 mm/year. On the other hand, the addition of antifreeze to ordinary water increases the corrosion potential of steel by more than 400 mV.

Keywords: corrosion and prevention, steel, copper, aluminum, corrosion inhibitor, anti-cooling

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Authors: Maria Lucia G. Lourenco, Keylla H. N. P. Pereira, Vivane Y. Hibaru, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Neonatal sepsis is a systemic response to the acute generalized infection caused by one or more bacterial agents, representing the main infectious cause of neonatal mortality in dogs during the first three weeks of life. This study aims to describe the incidence of sepsis in neonate dogs, as well as the main clinical signs and mortality rates. The study included 735 neonates admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, between January 2018 and November 2019. Seven hundred thirty-five neonates, 14% (98/703) presented neonatal sepsis. The main sources of infection for the neonates were intrauterine (72.5%, 71/98), lactogenic (13.2%, 13/98), umbilical (5.1%, 5/98) and unidentified sources (9.2%, 9/98). The main non-specific clinical signs observed in the newborns were weakness, depression, impaired or absent reflexes, hypothermia, hypoglycemia, dehydration, reduced muscle tonus and diarrhea. The newborns also manifested clinical signs of severe infection, such as hyperemia in the abdominal and anal regions, omphalitis, hematuria, abdomen and extremities with purplish-blue coloration necrosing injuries in the pads, bradycardia, dyspnea, epistaxis, hypotension and evolution to septic shock. Infections acquired during intrauterine life led to the onset of the clinical signs at the time of birth, with fast evolution during the first hours of life. On the other hand, infections acquired via milk or umbilical cord presented clinical signs later. The total mortality rate was 5.4% (38/703) and the mortality rate among the neonates with sepsis was 38.7% (38/98). The early mortality rate (0 to 2 days) accounted for 86.9% (33/38) and the late mortality rate (3 to 30 days) for 13.1% (5/38) of the deaths among the newborns with sepsis. The main bacterial agents observed were Staphylococcus spp., Streptococcus spp., Proteus spp. Mannheimia spp. and Escherichia coli. Neonatal sepsis evolves quickly and may lead to high mortality in a litter. The prognosis is usually favorable if the diagnosis is reached early and the antibiotic therapy instituted as soon as possible, even before the results of blood cultures and antibiograms. The therapeutic recommendations should meet the special physiological conditions of a neonate in terms of metabolism and excretion of medication. Therefore, it is of utmost importance that the veterinarian is knowledgeable regarding neonatology to provide effective intervention and improve the survival rates of these patients.

Keywords: Neonatal infection , bacteria, puppies, newborn

Procedia PDF Downloads 91

Authors: Eva Masson, Andrea Kübler

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Anxiety disorders (AD) are the most prevalent psychological disorders. However, far from most affected individuals are diagnosed and receive treatment. This gap is probably due to the diagnosis criteria, relying on symptoms (according to the DSM-5 definition) with no objective biomarker. Approach-avoidance conflict tasks are one common approach to simulate such disorders in a lab setting, with most of the paradigms focusing on the relationships between behavior and neurophysiology. Approach-avoidance conflict tasks typically place participants in a situation where they have to make a decision that leads to both positive and negative outcomes, thereby sending conflicting signals that trigger the Behavioral Inhibition System (BIS). Furthermore, behavioral validation of such paradigms adds credibility to the tasks – with overt conflict behavior, it is safer to assume that the task actually induced a conflict. Some of those tasks have linked asymmetrical frontal brain activity to induced conflicts and the BIS. However, there is currently no consensus for the direction of the frontal activation. The authors present here a modified version of the T-Maze paradigm, a motivational conflict desktop task, in which behavior is recorded simultaneously to the recording of high-density EEG (HD-EEG). Methods: In this within-subject design, HD-EEG and behavior of 35 healthy participants was recorded. EEG data was collected with a 128 channels sponge-based system. The motivational conflict desktop task consisted of three blocks of repeated trials. Each block was designed to record a slightly different behavioral pattern, to increase the chances of eliciting conflict. This variety of behavioral patterns was however similar enough to allow comparison of the number of trials categorized as ‘overt conflict’ between the blocks. Results: Overt conflict behavior was exhibited in all blocks, but always for under 10% of the trials, in average, in each block. However, changing the order of the paradigms successfully introduced a ‘reset’ of the conflict process, therefore providing more trials for analysis. As for the EEG correlates, the authors expect a different pattern for trials categorized as conflict, compared to the other ones. More specifically, we expect an elevated alpha frequency power in the left frontal electrodes at around 200ms post-cueing, compared to the right one (relative higher right frontal activity), followed by an inversion around 600ms later. Conclusion: With this comprehensive approach of a psychological mechanism, new evidence would be brought to the frontal asymmetry discussion, and its relationship with the BIS. Furthermore, with the present task focusing on a very particular type of motivational approach-avoidance conflict, it would open the door to further variations of the paradigm to introduce different kinds of conflicts involved in AD. Even though its application as a potential biomarker sounds difficult, because of the individual reliability of both the task and peak frequency in the alpha range, we hope to open the discussion for task robustness for neuromodulation and neurofeedback future applications.

Keywords: anxiety, approach-avoidance conflict, behavioral inhibition system, EEG

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Authors: Richa Mardianingrum, Srie R. N. Endah

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In this research, we have designed four isoniazid derivatives i.e., isonicotinohydrazide (1-isonicotinoyl semicarbazide, 1-thiosemi isonicotinoyl carbazide, N '-(1,3-dimethyl-1 h-pyrazole-5-carbonyl) isonicotino hydrazide, and N '-(1,2,3- 4-thiadiazole-carbonyl) isonicotinohydrazide. The docking and molecular dynamic have performed to them in order to study its interaction with Mycobacterium tuberculosis Enoyl-Acyl Carrier Protein Reductase (InhA). Based on this research, all of the compounds were predicted to have a stable interaction with Mycobacterium tuberculosis Enoyl-Acyl Carrier Protein Reductase (INHA) receptor, so they could be used as an anti-tuberculosis drug candidate.

Keywords: anti-tuberculosis, docking, Inhibin alpha subunit, InhA, inhibition, synthesis, isonicotinohydrazide

Procedia PDF Downloads 164

Authors: Georgia Saxami, Evangelia N. Kerezoudi, Evdokia K. Mitsou, Marigoula Vlassopoulou, Georgios Zervakis, Adamantini Kyriacou

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Autism Spectrum Disorder (ASD) is a complex group of developmental disorders of the brain, characterized by social and communication dysfunctions, stereotypes and repetitive behaviors. The potential interaction between gut microbiota (GM) and autism has not been fully elucidated. Children with autism often suffer gastrointestinal dysfunctions, while alterations or dysbiosis of GM have also been observed. Treatment with dietary components has been postulated to regulate GM and improve gastrointestinal symptoms, but there is a lack of evidence for such approaches in autism, especially for prebiotics. This study assessed the effects of Pleurotus eryngii mushroom (candidate prebiotic) and inulin (known prebiotic compound) on gut microbial composition, using faecal samples from autistic children in an in vitro batch culture fermentation system. Selected members of GM were enumerated at baseline (0 h) and after 24 h fermentation by quantitative PCR. After 24 h fermentation, inulin and P. eryngii mushroom induced a significant increase in total bacteria and Faecalibacterium prausnitzii compared to the negative control (gut microbiota of each autistic donor with no carbohydrate source), whereas both treatments induced a significant increase in levels of total bacteria, Bifidobacterium spp. and Prevotella spp. compared to baseline (t=0h) (p for all <0.05). Furthermore, this study evaluated the impact of fermentation supernatants (FSs), derived from P. eryngii mushroom or inulin, on the expression levels of tight junctions’ genes (zonulin-1, occludin and claudin-1) in Caco-2 cells stimulated by bacterial lipopolysaccharides (LPS). Pre-incubation of Caco-2 cells with FS from P. eryngii mushroom led to a significant increase in the expression levels of zonulin-1, occludin and claudin-1 genes compared to the untreated cells, the cells that were subjected to LPS and the cells that were challenged with FS from negative control (p for all <0.05). In addition, incubation with FS from P. eryngii mushroom led to the highest mean expression values for zonulin-1 and claudin-1 genes, which differed significantly compared to inulin (p for all <0.05). Overall, this research highlighted the beneficial in vitro effects of P. eryngii mushroom on the composition of GM of autistic children after 24 h of fermentation. Also, our data highlighted the potential preventive effect of P. eryngii FSs against dysregulation of the intestinal barrier, through upregulation of tight junctions’ genes associated with the integrity and function of the intestinal barrier. This research has been financed by "Supporting Researchers with Emphasis on Young Researchers - Round B", Operational Program "Human Resource Development, Education and Lifelong Learning."

Keywords: gut microbiota, intestinal barrier, autism spectrum disorders, Pleurotus Eryngii

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Authors: Yong-Cin Chen, Meng-Jiy Wang

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This study reports to prepare cuprous oxide (Cu2O) particles with different dimension and shape for evaluating the antibacterial applications. In the preparation of Cu2O, the surfactant, cetyltrimethylammonium bromide (CTAB), was used as templates to modulate the size of the prepared Cu2O particles. Furthermore, ammonia water was used for adjusting the pH environment that four different shapes of particles including cubic, spherical, octahedral, and star-like Cu2O were synthesized. The physical characteristics of Cu2O particles were evaluated by scanning electron microscope (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), UV/VIS spectrophotometer, and zeta potential meter/particle size analyzer (ZetaPALS). The resistance to bacteria was investigated against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) by applying the synthesized Cu2O particles that the qualitative analyses were facilitated by measuring the inhibition zone on Agar plate.

Keywords: copper oxide, cupric oxide, nanoparticles, antibacetrial

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Authors: Majid Forghani, Michael Khachay

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In genetics, the impact of neighboring amino acids on a target site is referred as the nearest-neighbor effect or simply neighbor effect. In this paper, a new method called wavelet particle decomposition representing the one-dimensional neighbor effect using wavelet packet decomposition is proposed. The main idea lies in known dependence of wavelet packet sub-bands on location and order of neighboring samples. The method decomposes the value of a signal sample into small values called particles that represent a part of the neighbor effect information. The results have shown that the information obtained from the particle decomposition can be used to create better model variables or features. As an example, the approach has been applied to improve the correlation of test and reference sequence distance with titer in the hemagglutination inhibition assay.

Keywords: antigenic variants, neighbor effect, wavelet packet, wavelet particle decomposition

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Authors: Patrícia Severino, Luciana Nalone, Daniele Martins, Marco Chaud, Classius Ferreira, Cristiane Bani, Ricardo Albuquerque

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Hydrogels produced with polymers have been used in the development of dressings for wound treatment and tissue revitalization. Our study on polymer nanocomposites containing silver nanoparticles shows antimicrobial activity and applications in wound healing. The effects are linked with the slow oxidation and Ag⁺ liberation to the biological environment. Furthermore, bacterial cell membrane penetration and metabolic disruption through cell cycle disarrangement also contribute to microbial cell death. The silver antimicrobial activity has been known for many years, and previous reports show that low silver concentrations are safe for human use. This work aims to develop a hydrogel using natural polymers (sodium alginate and gelatin) combined with silver nanoparticles for wound healing and with antimicrobial properties in cutaneous lesions. The hydrogel development utilized different sodium alginate and gelatin proportions (20:80, 50:50 and 80:20). The silver nanoparticles incorporation was evaluated at the concentrations of 1.0, 2.0 and 4.0 mM. The physico-chemical properties of the formulation were evaluated using ultraviolet-visible (UV-Vis) absorption spectroscopy, Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), and thermogravimetric (TG) analysis. The morphological characterization was made using transmission electron microscopy (TEM). Human fibroblast (L2929) viability assay was performed with a minimum inhibitory concentration (MIC) assessment as well as an in vivo cicatrizant test. The results suggested that sodium alginate and gelatin in the (80:20) proportion with 4 mM of AgNO₃ in the (UV-Vis) exhibited a better hydrogel formulation. The nanoparticle absorption spectra of this analysis showed a maximum band around 430 - 450 nm, which suggests a spheroidal form. The TG curve exhibited two weight loss events. DSC indicated one endothermic peak at 230-250 °C, due to sample fusion. The polymers acted as stabilizers of a nanoparticle, defining their size and shape. Human fibroblast viability assay L929 gave 105 % cell viability with a negative control, while gelatin presented 96% viability, alginate: gelatin (80:20) 96.66 %, and alginate 100.33 % viability. The sodium alginate:gelatin (80:20) exhibited significant antimicrobial activity, with minimal bacterial growth at a ratio of 1.06 mg.mL⁻¹ in Pseudomonas aeruginosa and 0.53 mg.mL⁻¹ in Staphylococcus aureus. The in vivo results showed a significant reduction in wound surface area. On the seventh day, the hydrogel-nanoparticle formulation reduced the total area of injury by 81.14 %, while control reached a 45.66 % reduction. The results suggest that silver-hydrogel nanoformulation exhibits potential for wound dressing therapeutics.

Keywords: nanocomposite, wound healing, hydrogel, silver nanoparticle

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