Search results for: ammonia oxidizing bacteria

Authors: Adel A. S. Al-Gheethi, M. O. Abdul-Monem

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Cellulase-producing bacteria were isolated from wastewater and sludge, and identified as Bacillus megaterium 1295S, Sporosarcina pasteurii 586S, Bacillus subtilis 117S, Burkholderia cepacia 120S and Staphylococcus xylosus 222W. Among bacteria, B. megaterium 1295S was the best cellulase producer under the catabolic repression and was therefore selected to study the factors affecting cellulase production. The optimum conditions for cellulase production were observed in CMC-Yeast Extract (CYE) agar medium (pH 6.5) inoculated with 0.4 mL of bacterial culture and incubated at 45˚C for 72 h. Twenty amino acids were introduced into the production medium as nitrogen source to investigate the production of cellulase in presence of amino acids in comparison to peptone (as an organic source) and sodium nitrate (as an inorganic source). The results found that the maximum production of cellulase was recorded at 50 ppm when L-hydroxy proline, L-arginine, glycine, L-histidine, L-leucine, DL-isoleucine, DL-β-phenylalanine were used as sole nitrogen sources and at 100 ppm when DL-threonine, L-ornithine 12.29, L-proline were used as sole nitrogen sources. The highest biomass yield was found when glycine 5 ppm and DL-serine 100 ppm used as a nitrogen source.

Keywords: CMCase, Bacillus megaterium 1295S, factors, amino acids

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Authors: Mirosław Krzemieniewski, Marcin Zieliński, Marcin Dębowski

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This publication presents a device for depolymerisation of plant substrates applicable to agricultural biogas plants and closed-chamber sewage treatment plants where sludge fermentation is bolstered with plant mass. The device consists of a tank with a cover equipped with a heating system, an inlet for the substrate, and an outlet for the depolymerised substrate. Within the tank, a magnet shaft encased in a spiral casing is attached, equipped on its upper end with an internal magnetic disc. A motoreducer is mounted on an external magnetic disc located on the centre of the cover. Depolymerisation of the plant substrate allows for substrate destruction at much lower power levels than by conventional means. The temperature within the reactor can be lowered by 40% in comparison to existing designs. During the depolymerisation process, free radicals are generated within the magnetic field, oxidizing the conditioned substrate and promoting biodegradation. Thus, the fermentation time in the fermenters is reduced by approximately 20%.

Keywords: depolymerisation, pre-treatment, biomass, fermentation

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Authors: Vaishnavi Unde, Srikanth Mutnuri

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Today dye stuff sector is one of the major chemical industries in India. Indigo is a blue coloured dye used all over the world in large quantity. The indigo dye produced and used in textile industries is synthetic having toxic effect, thus there is an increase in interest for natural dyes owing to the environmental concerns. The present study focuses on the use of a strain Pandoraea sp. isolated from garage soil, for the production of indigo in fed batch bioreactor. A comparative study between single phase and two phase production was carried out in this work. The blue colour produced during the experiments was analyzed using, TLC, UV-visible spectrophotometer and FTIR technique. The blue pigment was found to be indigo. The production of bio-indigo was done in a single phase fermentor carrying medium and substrate indole in dissolved form and was found to produce maximum of 0.041 g/L of indigo. Whereas there was an increase in production of indigo to 0.068 g/L in a two phase, water-silicone oil system. In this study the advantage of using second phase as silicone oil has enhanced the indigo production, as the second phase made the substrate available to the bacteria by increasing the surface area as well as it helped to prevent the inhibition effect of the high concentration of substrate, indole. The effect of single and two phases on the growth of bacteria was also studied.

Keywords: dyes, fed batch reactor, indole, Indigo

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Authors: Ramdani Nadia, Kheddaoui Abdelkrim, Nemmich Said, Tilmatine Amar

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The waste production nationwide is increasing every year, on account of therapid urbanization and growing populations, also consumption modes. Algerian political authorities have chosen Technical Landfill Centres (TLC) as a competitive and safe technique of waste management. However, storing these wastes in a bad way poses several environmental challenges, especially in the Department of Saïda, the latter have significant groundwaters. The major problem registered on this Landfill is the leachate resulting from the degradation of buried wastes which were disposed off the outside of the leachate basin and present a source of pollution for the local groundwaters by heavy metals and pathogenic germs. The present paper investigates the leachate treatment ozone process produced by Dielectric Barrier Discharge (DBD) under high potential. The experimental results obtained allowed us to show the efficiency of the treatment process by ozone based on the micro pollutant analysis (DCO, DBO5 , COT, heavy metals) and microbial analysis, after ozonation treatment. The results show that 80% of micro pollutants are eliminated and 100% destruction of all bacteria which reveals the high efficiency of the process.

Keywords: landfill, leachate, treatment, ozone, polluants, bacteria, micropolluant

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Authors: Said Abdirasak Abidrahman, Ibrahim Mohamed

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Background: Urinary Tract Infections (UTIs) are the commonest infections described among diabetes mellitus patients. More often, empirical antimicrobial therapy is initiated before the laboratory results are made available with minimal treatment success. The knowledge of the etiology and antibiotic susceptibility patterns of the organisms causing urinary tract infections among diabetes mellitus patients remains scarce, despite its vitality. This study sought to determine the prevalence, bacteria species, and drug susceptibility patterns of common causes of urinary tract infections among diabetes mellitus patients attending Bosaso health centers. Materials and methods: We conducted a cross-sectional study involving adult diabetic patients at Bosaso health centers between the months of May and July 2020. Laboratory assay of mid-stream urine samples was done to isolate bacteria causes of UTIs. These were biochemically identified using Gram stain, Kligler iron agar (KIA), Indole test, citrate, urea, coagulase, catalase, motility agar, and lysine iron agar. Their antibiotic susceptibility pattern for the isolated organisms was made for Ampicillin 10μg, Ciprofloxacin 5μg, Cotrimoxazole 25μg, Gentamycin 10μg, Ceftriaxone 10μg, and determined using the Kirby Bauer Disc Diffusion method. Results: Of 177 participants, 69 (39.0%) were males and 108 (61.0%) were females. Their mean age was 33.1 years (range; 18-67 years). Of these, 14.7% (26/177) of the samples revealed significant growth (>= 105 CFU/mL) giving a prevalence of 14.9 % (95% CI: 10.6 to 16.3). The organisms isolated were Escherichia coli -50% (N=13), Klebsiella pneumonia 30.8% (N=8), Staphylococcus aureus 15.4% (N=4), and unidentified organism 3.8% (N=1), and these were associated with such socio-demographic factors like history of catheterization and sexual activity. Antibiotic susceptibility to the commonly used agents for treating UTIs indicated higher sensitivity to Gentamicin and Ceftriaxone.

Keywords: antimicrobials, bacteria, urinary tract infections, diabetes

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Authors: Lali Kutateladze, Tamar Urushadze, Tamar Dudauri, Besarion Metreveli, Nino Zakariashvili, Izolda Khokhashvili, Maya Jobava

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Lignocellulosic waste streams from agriculture, paper and wood industry are renewable, plentiful and low-cost raw materials that can be used for large-scale production of liquid and gaseous biofuels. As opposed to prevailing multi-stage biotechnological processes developed for bioconversion of cellulosic substrates to ethanol where high-cost cellulase preparations are used, Consolidated Bioprocessing (CBP) offers to accomplish cellulose and xylan hydrolysis followed by fermentation of both C6 and C5 sugars to ethanol in a single-stage process. Syntrophic microbial consortium comprising of anaerobic, thermophilic, cellulolytic, and saccharolytic bacteria in the genus Clostridia with improved ethanol productivity and high tolerance to fermentation end-products had been proposed for achieving CBP. 65 new strains of anaerobic thermophilic cellulolytic and saccharolytic Clostridia were isolated from different wetlands and hot springs in Georgia. Using new isolates, fermentation of mechanically pretreated wheat straw and corn stalks was done under oxygen-free nitrogen environment in thermophilic conditions (T=550C) and pH 7.1. Process duration was 120 hours. Liquid and gaseous products of fermentation were analyzed on a daily basis using Perkin-Elmer gas chromatographs with flame ionization and thermal detectors. Residual cellulose, xylan, xylose, and glucose were determined using standard methods. Cellulolytic and saccharolytic bacteria strains degraded mechanically pretreated herbaceous cellulosic wastes and fermented glucose and xylose to ethanol, acetic acid and gaseous products like hydrogen and CO2. Specifically, maximum yield of ethanol was reached at 96 h of fermentation and varied between 2.9 – 3.2 g/ 10 g of substrate. The content of acetic acid didn’t exceed 0.35 g/l. Other volatile fatty acids were detected in trace quantities.

Keywords: anaerobic bacteria, cellulosic wastes, Clostridia sp, ethanol

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Authors: Nomakholwa F. Stokwe, Antoinette P. Malan

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Woolly apple aphid (WAA), Eriosoma lanigerum, is an important pest of apples worldwide. The aphid feeds above ground on buds and leaf axils and the roots of apple trees. Entomopathogenic nematodes (EPNs) of the two families, Steinernematidae and Heterorhabditidae, and their symbiotic bacteria have generated extensive interest as inundative applied biological control agents of insects. With the development of the resistance of WAA to chemicals, export restrictions, and the inability of parasitoids to control the aphid successfully early in the season, considering EPNs as an alternative biocontrol agent is important. Seven EPN species were tested for their pathogenicity against WAA. Laboratory bioassays identified S. yirgalemense and H. zealandica as being the most virulent against the subterranean stage of the WAA, with a mortality rate of 48% and 38%, respectively. Studies on the effect of WAA size showed that the last instar is most susceptible to infection, whereas smaller instars appear to be too small for nematode penetration and infection. Neither increasing the exposure period of the aphids nor increasing the nematode concentration affected the infection rate positively. The haemolymph of WAA showed an inhibitory effect on the development of the symbiotic bacteria, preventing the completion of the nematode’s life cycle.

Keywords: apples, biocontrol, entomopathogenic nematodes, woolly apple aphid

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Authors: Zeinab Hajifathali, Moghan Amirhosseinian

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This study had two main aims: firstly, to determine how the individual substitution of CaO/MgO and CaO/SrO can affect the in vitro bioactivity of sol-gel derived substituted 58S bioactive glass (BG) and secondly to introduce a composition in the 60SiO₂–(36-x)CaO–4P₂O₅–(x)MgO and 60SiO₂–(36-x)CaO–4P₂O₅–(x)SrO quaternary systems (where x= 0, 5, 10 mol.%) with enhanced biocompatibility, alkaline phosphatase (ALP) activity, and more efficient antibacterial activity against MRSA bacteria. Results showed that both magnesium-substituted bioactive glasses (M-BGs) and strontium- substituted bioactive glasses (S-BGs) retarded the Hydroxyapatite (HA) formation. Meanwhile, magnesium had more pronounced effect. The 3-(4, 5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and ALP assays revealed that the presence of moderate amount (5 mol%) of Mg and Sr had a stimulating effect on increasing of both proliferation and differentiation of MC3T3-E1 cells. Live dead and Dapi/actin staining revealed both substitution of CaO/MgO and CaO/SrO resulted in more biocompatibility and stimulation potential of the MC3T3 cells compared with control. Taken together, among all of the synthesized magnesium substituted (MBGs) and strontium substituted (SBGs), the sample 58- BG with 5 mol% CaO/MgO substitution (BG-5M) was considered as a multifunctional biomaterial in bone tissue regeneration field with enhanced biocompatibility, ALP activity as well as the highest antibacterial efficiency against methicillin-resistant Staphylococcus aureus (MRSA) bacteria.

Keywords: apatite, alkaline earth, bioactivity, biomedical applications, Sol-gel

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Authors: Z. Yousefniayejahr, S. Bolognini, A. Bonini, C. Campobasso, N. Poma, F. Vivaldi, M. Di Luca, A. Tavanti, F. Di Francesco

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Pathogenic bacteria represent a threat to healthcare systems and the food industry because their rapid detection remains challenging. Electrochemical biosensors are gaining prominence as a novel technology for the detection of pathogens due to intrinsic features such as low cost, rapid response time, and portability, which make them a valuable alternative to traditional methodologies. These sensors use biorecognition elements that are crucial for the identification of specific bacteria. In this context, bacteriophages are promising tools for their inherent high selectivity towards bacterial hosts, which is of fundamental importance when detecting bacterial pathogens in complex biological samples. In this study, we present the development of a low-cost and portable sensor based on the Zeno phage for the rapid detection of Staphylococcus aureus. Screen-printed gold electrodes functionalized with the Zeno phage were used, and electrochemical impedance spectroscopy was applied to evaluate the change of the charge transfer resistance (Rct) as a result of the interaction with S. aureus MRSA ATCC 43300. The phage-based biosensor showed a linear range from 101 to 104 CFU/mL with a 20-minute response time and a limit of detection (LOD) of 1.2 CFU/mL under physiological conditions. The biosensor’s ability to recognize various strains of staphylococci was also successfully demonstrated in the presence of clinical isolates collected from different geographic areas. Assays using S. epidermidis were also carried out to verify the species-specificity of the phage sensor. We only observed a remarkable change of the Rct in the presence of the target S. aureus bacteria, while no substantial binding to S. epidermidis occurred. This confirmed that the Zeno phage sensor only targets S. aureus species within the genus Staphylococcus. In addition, the biosensor's specificity with respect to other bacterial species, including gram-positive bacteria like Enterococcus faecium and the gram-negative bacterium Pseudomonas aeruginosa, was evaluated, and a non-significant impedimetric signal was observed. Notably, the biosensor successfully identified S. aureus bacterial cells in a complex matrix such as a nasal swab, opening the possibility of its use in a real-case scenario. We diluted different concentrations of S. aureus from 108 to 100 CFU/mL with a ratio of 1:10 in the nasal swap matrices collected from healthy donors. Three different sensors were applied to measure various concentrations of bacteria. Our sensor indicated high selectivity to detect S. aureus in biological matrices compared to time-consuming traditional methods, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and radioimmunoassay (RIA), etc. With the aim to study the possibility to use this biosensor to address the challenge associated to pathogen detection, ongoing research is focused on the assessment of the biosensor’s analytical performances in different biological samples and the discovery of new phage bioreceptors.

Keywords: electrochemical impedance spectroscopy, bacteriophage, biosensor, Staphylococcus aureus

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Authors: Chidambaram Thamaraiselvan, Carlos Dosoretz

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This work will explore the influence of low voltage electric field, both alternating (AC) and direct (DC) currents, on biofouling control to highly electrically conductive self-supporting carbon nanotubes (CNT) membranes at conditions which encourage bacterial growth. A mutant strain of Pseudomonas putida S12 was used a model bacterium. The antibiofouling studies were performed with flow-through mode connecting an electric circuit in resistive mode. Major emphasis was placed on AC due to its ability of repulsing and inactivating bacteria. The observations indicate that an AC potential >1500 mV, 1 kHz frequency, 100 Ω external resistance on ground side and pulse wave above the offset (+0.45) almost completely prevented attachment of bacteria (>98.5%) and bacterial inactivation (95.3±2.5%). Findings suggest that at the conditions applied, direct electron transfer might be dominant in a decrease of cell viability. AC resulted more effective than DC, both in terms of biofouling reduction compared to cathodic DC and in terms of cell inactivation compared to anodic DC. This electrically polarized CNT membranes offer a viable antibiofouling strategy to hinder biofouling and simplify membrane care during filtration.

Keywords: bacterial attachment, biofouling control, low electric potential, water treatment

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Authors: Ilma Robo, Manola Kelmendi, Kleves Elezi, Nevila Alliu

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The most important stage in deciding whether to re-treat or not endodontically is to find the reason for the clinical in-success. Therefore, endodontic re-treatment aims to eliminate the etiology of the pathology, where the main ones are the bacteria remaining in the inter-radicular spaces or the presence of other irritants that can be not only bacterial toxins but also the elements that keep the batteries fixed or extra-canal toxins such as extraction outside the apex of the canal filling. Shortcomings of endodontic treatment can be corrected, if possible, only with endodontic re-treatment that is initially attempted orthograde, and if clinical endodontic success is not achieved again, it can be performed retrograde or surgically. The elements that do not help in this direction are the anatomical deformations in the canal network of the tooth roots, in the presence of the delta at the apex of the tooth root, in the isthmuses present, all of which can be explained by the endodontic canal anatomical morphology. Actually, even if the causative endodontic bacteria remains isolated and without an exit in the healthy periodontal tissues, then this can also be a clinical endodontic success, regardless of the fact that the endodontic isolation occurred only in the exits such as the apex or the accessory canals. Clinical endodontic in-success occurs only when bacterial residues emerge or provide an exit in the healthy periradicular tissues or along the entire length of the canal where the accessory canals exit.

Keywords: endodontic success, E. foecalis, nanoparticles, laser diode, antibacterial, antiseptic

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Authors: M. T. Darzi, M. Shirkhodaei, M. R. Haj Seyed Hadi

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The main objective of this study was to determine the effects of organic fertilizer and azotobacter and azospirillum bacteria on concentration and composition of essential oil in the coriander essential oil content, essential oil yield, linalool percent, alpha pinene percent and cymene percent in essential oil. The experiment was carried out as factorial experiment in the base of randomized complete blocks design with eight treatments and three replications at research field of Agriculture Company of Ran in Firouzkuh of iran in 2012. The factors were Vermicompost in four levels (0, 3, 6 and 9 ton/ha) and biofertilizer, mixture of Azotobacter chroococcum and Azospirillum lipoferum in two levels (non-inoculated and inoculated seeds). The present results have shown that vermicompost had significant effects on evaluated traits except linalool percent in essential oil, as the highest essential oil content, essential oil yield and alpha pinene percent in essential were obtained after applying 6 ton/ha vermicompost. The minimum cymene percent in essential oil were obtained after applying 6 ton/ha vermicompost. Biofertilizer also showed significant effects on essential oil yield only. The highest essential oil yield were obtained by using the biofertilizer (inoculated seeds).

Keywords: coriander, vermicompost, biofertilizer, essential oil

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Authors: Muhammad Adeel Arshad, Shaukat Ali Bhatti, Faiz-ul Hassan

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Manipulating microbial ecosystem in the rumen is considered as an important strategy to optimize production efficiency in ruminants. In the past, antibiotics and synthetic chemical compounds have been used for the manipulation of rumen fermentation. However, since the non-therapeutic use of antibiotics has been banned, efforts are being focused to search out safe alternative products. In tropics, crop residues and forage grazing are major dietary sources for ruminants. Poor digestibility and utilization of these feedstuffs by animals is a limiting factor to exploit the full potential of ruminants in this area. Hence, there is a need to enhance the utilization of these available feeding resources. One of the potential strategies in this regard is the use of direct-fed microbes. Bacteria and fungi are mostly used as direct-fed microbes to improve animal health and productivity. Commonly used bacterial species include lactic acid-producing and utilizing bacteria (Lactobacillus, Streptococcus, Enterococcus, Bifidobacterium, and Bacillus) and fungal species of yeast are Saccharomyces and Aspergillus. Direct-fed microbes modulate microbial balance in the gastrointestinal tract through the competitive exclusion of pathogenic species and favoring beneficial microbes. Improvement in weight gain and feed efficiency has been observed as a result of feeding direct-fed bacteria. The use of fungi as a direct-fed microbe may prevent excessive production of lactate and harmful oxygen in the rumen leading to better feed digestibility. However, the mechanistic mode of action for bacterial or fungal direct-fed microbes has not been established yet. Various reports have confirmed an increase in dry matter intake, milk yield, and milk contents in response to the administration of direct-fed microbes. However, the application of a direct-fed microbe has shown variable responses mainly attributed to dosages and strains of microbes. Nonetheless, it is concluded that the inclusion of direct-fed microbes may mediate the rumen ecosystem to manage lactic acid production and utilization in both clinical and sub-acute rumen acidosis.

Keywords: microbes, roughages, rumen, feed efficiency, production, fermentation

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Authors: N. Chikhi-Chorfi, L. Arbia, S. Zenia, H.Lounici

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Opuntia ficus-indica belongs to the Cactaceae family. The cactus is mainly cultivated for its fruit (prickly pear) that, eaten after pealing, is sweet and juicy, and rich in nutritional compounds, such as ascorbic acid and polyphenols. Different parts of O. ficus-indica are used in the traditional medicine of several countries: the cladodes are utilized to reduce serum cholesterol level and blood pressure, for treatment of ulcers, rheumatic pain, wounds, fatigue, capillary fragility, and liver conditions. This original study, investigate the effect of polyphenols of O. ficus indica (cactus) cladodes against periodontal bacteria collected from patients with periodontitis. The quantitative analysis of total polyphenols (TPP) was determined with Follin-Ciocalteu method. Different concentrations of extracts of O. ficus indica were tested by the disk method on two bacterial strains: Porphyromonas gingivalis and Prevotella intermedia responsible for periodontal disease. The results showed a good correlation between the concentration of total polyphenols and the antibacterial activity of the extracts of Opuntia ficus indica against P. gingivalis and P. intermedia with R² = 0.94 and R² = 0.90 respectively. This observation suggests that these extracts could be used in the treatment and prevention of periodontitis.

Keywords: periodontal disease, P. gingivalis, P. intermedia, polyphenols, Opuntia ficus indica

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Authors: Shih-Huang Pan, Tsuyoshi Miyazaki, Minoru Otani, Santhanamoorthi Nachimuthu, Jyh-Chiang Jiang

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Ammonia (NH₃) is crucial in renewable energy and agriculture, yet its traditional production via the Haber-Bosch process faces challenges due to the inherent inertness of nitrogen (N₂) and the need for high temperatures and pressures. The electrocatalytic nitrogen reduction (ENRR) presents a more sustainable option, functioning at ambient conditions. However, its advancement is limited by selectivity and efficiency challenges due to the competing hydrogen evolution reaction (HER). The critical roles of protonation of N-species and HER highlight the necessity of selecting optimal catalysts and solvents to enhance ENRR performance. Notably, transition metal oxides, with their adjustable electronic states and excellent chemical and thermal stability, have shown promising ENRR characteristics. In this study, we use density functional theory (DFT) methods to investigate the ENRR mechanisms on IrO₂ (110), a material known for its tunable electronic properties and exceptional chemical and thermal stability. Employing the constant electrode potential (CEP) model, where the electrode - electrolyte interface is treated as a polarizable continuum with implicit solvation, and adjusting electron counts to equalize work functions in the grand canonical ensemble, we further incorporate the advanced 3D Reference Interaction Site Model (3D-RISM) to accurately determine the ENRR limiting potential across various solvents and pH conditions. Our findings reveal that the limiting potential for ENRR on IrO₂ (110) is significantly more favorable than for HER, highlighting the efficiency of the IrO₂ catalyst for converting N₂ to NH₃. This is supported by the optimal *NH₃ desorption energy on IrO₂, which enhances the overall reaction efficiency. Microkinetic simulations further predict a promising NH₃ production rate, even at the solution's boiling point¸ reinforcing the catalytic viability of IrO₂ (110). This comprehensive approach provides an atomic-level understanding of the electrode-electrolyte interface in ENRR, demonstrating the practical application of IrO₂ in electrochemical catalysis. The findings provide a foundation for developing more efficient and selective catalytic strategies, potentially revolutionizing industrial NH₃ production.

Keywords: density functional theory, electrocatalyst, nitrogen reduction reaction, electrochemistry

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Authors: Abdallah Abusam, Andrzej Mydlarczyk, Fadila Al-Salameen, Moh Elmuntasir Ahmed

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Treated municipal wastewater produced in Kuwait is used mainly in agricultural and greenery landscape irrigations. However, there are strong doubts that severe sludge bulking and foaming problems, particularly during winter seasons, may render the treated wastewater to be unsuitable for irrigation purposes. To assess the impact of sludge bulking and foaming problems on the quality of treated effluents, samples were collected weekly for nine months (January to September 2014) from the secondary effluents, tertiary effluents and sludge-mixed liquor streams of the two plants that severely suffer from sludge bulking and foaming problems. Dominant filamentous bacteria were identified and quantified using a molecular method called VIT (Vermicon Identification Technology). Quality of the treated effluents was determined according to water and wastewater standard methods. Obtained results were then statistically analyzed and compared to irrigation water standards. Statistical results indicated that secondary effluents were greatly impacted by sludge bulking and foaming problems, while tertiary effluents were slightly affected. This finding highlights the importance of having tertiary treatment units in plants that encountering sludge bulking and foaming problems.

Keywords: agriculture, filamentous bacteria, reclamation, reuse, wastewater

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Authors: Goksen Arik, Mihriban Korukluoglu

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Lactic acid bacteria (LAB) and some yeast species are common microorganisms found in dairy products and most of them are responsible for the fermentation of foods. Such cultures are isolated and used as a starter culture in the food industry because of providing standardisation of the final product during the food processing. Choice of starter culture is the most important step for the production of fermented food. Isolated LAB and yeast cultures which have the ability to create a biofilm layer can be preferred as a starter in the food industry. The biofilm formation could be beneficial to extend the period of usage time of microorganisms as a starter. On the other hand, it is an undesirable property in pathogens, since biofilm structure allows a microorganism become more resistant to stress conditions such as antibiotic presence. It is thought that the resistance mechanism could be turned into an advantage by promoting the effective microorganisms which are used in the food industry as starter culture and also which have potential to stimulate the gastrointestinal system. Development of the biofilm layer is observed in some LAB and yeast strains. The resistance could make LAB and yeast strains dominant microflora in the human gastrointestinal system; thus, competition against pathogen microorganisms can be provided more easily. Based on this circumstance, in the study, 10 LAB and 10 yeast strains were isolated from various dairy products, such as cheese, yoghurt, kefir, and cream. Samples were obtained from farmer markets and bazaars in Bursa, Turkey. As a part of this research, all isolated strains were identified and their ability of biofilm formation was detected with two different methods and compared with each other. The first goal of this research was to determine whether isolates have the potential for biofilm production, and the second was to compare the validity of two different methods, which are known as “Tube method” and “96-well plate-based method”. This study may offer an insight into developing a point of view about biofilm formation and its beneficial properties in LAB and yeast cultures used as a starter in the food industry.

Keywords: biofilm, dairy products, lactic acid bacteria, yeast

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Authors: I. Nanako, Mariko Era, Hiroshi Morita

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Objectives: Japanese uses TATAMI rather than flooring at home. Igusa ( Juncus effuses var. decipiens ), which is commonly known in the forms of TATAMI. Juncus spp. grow at a relatively high humidity area (Japan, China and Southeast Asia ). Yatsushiro region in the southern part of Kumamoto prefecture is major produing area of Igusa. Igusa found to have honeycomb structure and was also shown to have the ability to control humidity. And Igusa has been used as a medicinal herb for diuretic and antiphlogistic agent. In previous study, we investigated antimicrobial effects of Igusa, and showed high antimicrobial activity against food poisoning bacteria. Therefore, the food trays blended Igusa can be kept clean by antimicrobial activity of Igusa. We focus on ‘Igusa foam materials’. In this study, we investigated the antibacterial and antifungal activity of Igusa, and new application to foam materials for food industry. Materials and method: We used Igusa foam materials (3 × 3 × 3 cm) as a sample. We set about fifteen types of samples combined with a commercial antibacterial agent A, a commercial antibacterial agent B, potassium laurate (C12K) and a commercial antifungal agent C, a commercial antifungal agent D and a commercial antifungal agent E. We selected four bacteria strains (Escherichia coli NBRC 3972, Staphylococus aureus NBRC 12732, Salmonella typhimurium NBRC 13245, Bacillus subtilis NBRC 3335 ) and three fungus strains (Penicillium pinophilum NBRC 6345, Cladosporium cladosporioides NBRC 30314, Aspergillus oryzae NBRC 5238 ). The fungus was cultured at 30 °C on Igusa foam materials after inoculation of the fungus for fourteen days. The bacteria was cultured at 30 °C on Igusa foam materials after inoculation of the bacteria for three days. And the Igusa foam materials were washed with 10 mL normal saline after three days. The normal saline washed Igusa foam materials plated the NA medium. After, It was cultured at 30 °C and used colony counting method. Result and Conclusion: The fifteen types of sample of Igusa foam materials had antifungal activity against C. cladosporioides, A. oryzae and P. pinophilum for fourteen days. The four types of sample contained potassium laurate and antibacterial agent A, sample contained antibacterial agent B and antifungal agent D, sample contained A and antifungal agent E, sample contained B and E had antibacterial activity against B. subtilis. The three types of sample contained potassium laurate and A, sample contained B and D, sample contained A and E had antibacterial activity against S. typhimurium. The five types of sample contained potassium laurate and A, sample contained B and D, sample contained A and E, sample contained B and E, sample contained B and antifungal agent C had antibacterial activity against E. coli and S. aureus. These results indicate that Igusa of Igusa foam materials had high antifungal activity. In addition, Igusa foam materials combined with a commercial antibacterial agent had antibacterial activity. In the future, we consider that use of Igusa foam materials may be spread from food industry.

Keywords: antibacterial, antifungal, foam materials, Igusa

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Authors: Sahira Nsayef Muslim, Israa M. S. Al-Kadmy, Nadheema Hammood Hussein, Alaa Naseer Mohammed Ali, Buthainah Mohammed Taha, Rayim Sabah Abbood, Sarah Naji Aziz

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A novel strain of Bacillus licheniformis isolated from spoilt cucumber and pepper samples have the ability to produce the chitosanase enzyme when grown on chitosan substrate. Chitosanase was purified to homogeneity with a recovery yield of 35.71% and 5.5 fold of purification by using ammonium sulfate at 45% saturation followed by ion exchange chromatography on DEAE-cellulose column and gel filtration chromatography on Sephadex G-100 column. The purified chitosanase inhibited the biofilm formation ability for all Gram-negative and Gram-positive biofilm-forming bacteria (biofilm producers) after using Congo Red agar and Microtiter plates methods. Highly antibiofilm of chitosanase recorded against Pseudomonas aeruginosa followed by Klebsiella pneumoniae with reduction of biofilm formation ratio to 22 and 29%, respectively compared with (100)% of control. Thus, chitosanase has promising benefit as antibiofilm agent against biofilm forming pathogenic bacteria and has promising application as alternative antibiofilm agents to combat the growing number of multidrug-resistant pathogen-associated infections, especially in situation where biofilms are involved.

Keywords: chitosanase, Bacillus licheniformis, vegetables, biofilm

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Authors: Salwa A. H. Hamdi, Maha A. M. El-Shazly, Mona Fathi Fol, Hanan S. Mossalem, Mosad A. Ghareeb, Amina M. Ibrahim

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One of the most distinctive and defining features of cephalopods squid, cuttlefish, and Octopus is their inking behavior. Their ink, which is blackened by melanin but also contains other constituents, has been used by humans in various ways for millennia. The present study aims to investigate the chemical profiling of the Octopus vulgaris ink extract and to evaluate its antioxidant, antimicrobial, and anti-schistosomal activities. The present results showed that GC-MS examination of Octopus vulgaris ink comprises 21 compounds. The main detected compounds are (E)-1, 2, 3, 4-Tetra (4-phenylphenyl)-2-butene-1,4-dione, Lipo-3-episapelin A, and 5,10-Dihexyltetrabenzoporphyrin. Results showed that the octopus ink had antioxidant capacity and the capability to mask DPPH free radicals in comparison with ascorbic acid. Octopus Vulgaris ink extract had inhibitory action against three gram-positive bacteria, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis, and three gram-negative bacteria, Neisseria gonorrhoeae, Escherichia coli, and Pseudomonas aeuroginosa. Additionally, the extracted ink revealed antifungal activity against Aspergillus flavus and yeast as Candida albicans. The obtained data indicated the effectiveness of ink extract in pharmaceutical industries as an antioxidant, antimicrobial and antischistosomicidal

Keywords: antimicrobial, antioxidant, ink, octopus vulgaris

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Authors: Seyed Hassan Moosa-kazemi, Jalal Mohammadi Soleimani, Hassan Vatandoost, Mohammad Hassan Shirazi, Sara Hajikhani, Roonak Bakhtiari, Morteza Akbari, Siamak Hydarzadeh

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Malaria is an infectious disease and considered most important health problems in the southeast of Iran. Iran is elimination malaria phase and new tool need to vector control. Paraterasgenesis is a new way to cut of life cycle of the malaria parasite. In this study, the microflora of the surface and gut of various stages of Anopheles fluviatilis James as one of the important malaria vector was studied using biochemical and molecular techniques during 2013-2014. Twelve bacteria species were found including; Providencia rettgeri, Morganella morganii, Enterobacter aerogenes, Pseudomonas oryzihabitans, Citrobacter braakii، Citrobacter freundii، Aeromonas hydrophila، Klebsiella oxytoca, Citrobacter koseri, Serratia fonticola، Enterobacter sakazakii and Yersinia pseudotuberculosis. The species of Alcaligenes faecalis, Providencia vermicola and Enterobacter hormaechei were identified in various stages of the vector and confirmed by biochemical and molecular techniques. We found Providencia rettgeri proper candidate for paratransgenesis.

Keywords: Anopheles fluviatilis, bacteria, malaria, Paraterasgenesis, Southern Iran

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Authors: Evelyn Osehontue Uroro, Richard Bright, Jing Yang Quek, Krasimir Vasilev

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Bacterial infections have been a challenge both in the public and private sectors. The colonization of bacteria often occurs in medical devices such as catheters, heart valves, respirators, and orthopaedic implants. When biomedical devices are inserted into patients, the deposition of macromolecules such as fibrinogen and immunoglobin on their surfaces makes it easier for them to be prone to bacteria colonization leading to the formation of biofilms. The formation of biofilms on medical devices has led to a series of device-related infections which are usually difficult to eradicate and sometimes cause the death of patients. These infections require surgical replacements along with prolonged antibiotic therapy, which would incur additional health costs. It is, therefore, necessary to prevent device-related infections by inhibiting the formation of biofilms using intelligent technology. Antibiotic resistance of bacteria is also a major threat due to overuse. Different antimicrobial agents have been applied to microbial infections. They include conventional antibiotics like rifampicin. The use of conventional antibiotics like rifampicin has raised concerns as some have been found to have hepatic and nephrotoxic effects due to overuse. Hence, there is also a need for proper delivery of these antibiotics. Different techniques have been developed to encapsulate and slowly release antimicrobial agents, thus reducing host cytotoxicity. Examples of delivery systems are solid lipid nanoparticles, hydrogels, micelles, and polymeric nanoparticles. The different ways by which drugs are released from polymeric nanoparticles include diffusion-based release, elution-based release, and chemical/stimuli-responsive release. Polymeric nanoparticles have gained a lot of research interest as they are basically made from biodegradable polymers. An example of such a biodegradable polymer is polycaprolactone (PCL). PCL degrades slowly by hydrolysis but is often sensitive and responsive to stimuli like enzymes to release encapsulants for antimicrobial therapy. This study presents the synthesis of PCL nanoparticles loaded with rifampicin and the on-demand release of rifampicin for treating staphylococcus aureus infections.

Keywords: enzyme, Staphylococcus aureus, PCL, rifampicin

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Authors: Hai Chi, Ibrahim Mehmeti, Kirill Ovchinnikov, Hegle Holo, Ingolf F. Nes, Dzung B. Diep

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By using a panel of multiple indicator strains of different bacterial species and genera, we screened a large collection of bacterial isolates (over 1800 isolates) derived from raw milk, for bacteriocin producers with broad inhibition spectra (BIS). Fourteen isolates with BIS were identified, and by 16S rDNA sequencing they were found to belong to Lactococcus garvieae (10 isolates) and Enterococcus feacalis (4 isolates). Further analysis of the ten L. garvieae isolates revealed that they were very similar, if not identical, to each other in metabolic and genetic terms: they had the same fermentation profile on different types of sugars, repetitive sequence-based PCR (rep-PCR) DNA pattern as well as they all had the same inhibition profile towards over 50 isolates of different species. The bacteriocin activity from one of the L. garvieae isolates was assessed further. The bacteriocin which was termed garvicin KS, was found to be heatstable and proteinase-labile and its inhibition spectrum contained many distantly related genera of Firmicutes, comprising most lactic acid bacteria (LAB) as well as problematic species of Bacillus, Listeria, Streptococcus and Staphylococcus and their antibiotic resistant derivatives (e.g. VRE, MRSA). Taken together, the results indicate that this is a potent bacteriocin from L. garvieae and that its very broad inhibition spectrum can be a very useful property for use in food preservation as well as in infection treatments caused by gram-positive pathogens and their antibiotic-derivatives.

Keywords: bacteriocin, lactic acid bacteria, Lactococcus garvieae, antibiotics resistance

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Authors: Stephen Olusanmi Akintayo

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Access to safe drinking water remains a major challenge in Nigeria, and where available, the quality of the water is often in doubt. An alternative to the inadequate clean drinking water is being found in treated drinking water packaged in electrically heated sealed nylon and commonly referred to as “sachet water”. “Sachet water” is a common thing in Nigeria as the selling price is within the reach of members of the low socio- economic class and the setting up of a production unit does not require huge capital input. The bacteriological quality of selected “sachet water” stored at room temperature over a period of 56 days was determined to evaluate the safety of the sachet drinking water. Test for the detection of coliform bacteria was performed, and the result showed no coliform bacteria that indicates the absence of fecal contamination throughout 56 days. Heterotrophic plate count (HPC) was done at an interval 14 days, and the samples showed HPC between 0 cfu/mL and 64 cfu/mL. The highest count was observed on day 1. The count decreased between day 1 and 28, while no growths were observed between day 42 and 56. The decrease in HPC suggested the presence of residual disinfectant in the water. The organisms isolated were identified as Staphylococcus epidermis and S. aureus. The presence of these microorganisms in sachet water is indicative for contamination during processing and handling.

Keywords: coliform, heterotrophic plate count, sachet water, Staphyloccocus aureus, Staphyloccocus epidermidis

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Authors: Piña-Ronces Laura Gabriela, Reyes-Escogido María de Lourdes

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Exist a large variability in Exopolysaccharides (EPS) produced by LAB depending on carbon source, they have multiple applications in food industry mainly, but they have become important for the health. In this study, we identified EPS-producing strains belonging to the BAL group; they were previously isolated from humans. After that, we extracted and evaluated the antioxidant activity of EPS produced by all strains. Antioxidant activity was determined by DPPH method using ascorbic acid as standard for both comparison and quantification. 31 strains (51.66 %) produced EPS at concentrations between 451 and 1.561 mg/l, 16 of EPS extracted showed antioxidant effect superior to ascorbic acid at the same concentrations. EPS-producing strains were L. plantarum, L. sp and L. fermentum corresponding to Lactobacillus genus and, E. faecium, E. durans, and E. hirae of Enterococcus genus. Antioxidant activity showed by EPS from 3 strains of L. plantarum and 3 strains of E. faecium was different into specie, while the antioxidant activity determined for EPS obtained from the other strains did not show difference at specie level, but was superior to ascorbic acid. EPS produced by L. plantarum and E. hirae had the best activity, it could be considerate for selection them as a possible new alternative for therapy or treatment of diseases related whit oxidative stress. Further studies about biological functions of EPS have to be conducted for new applications in health.

Keywords: oxidative stress, lactic acid bacteria, exopolysaccharides, antioxidant activity

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Authors: Satyendra Mourya, Jyoti Jaiswal, Gaurav Malik, Brijesh Kumar, Ramesh Chandra

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Present work describes the fabrication and sensing characteristics of the Pd-Pt decorated nanostructured silicon carbide (SiC) thin films on anodized porous silicon (PSi) substrate by RF magnetron sputtering. The gas sensing performance of Pd-Pt/SiC/PSi sensing electrode towards H2 gas under low (10–400 ppm) detection limit and high operating temperature regime (25–600 °C) were studied in detail. The chemiresistive sensor exhibited high selectivity, good sensing response, fast response/recovery time with excellent stability towards H2 at high temperature. The selectivity measurement of the sensing electrode was done towards different oxidizing and reducing gases and proposed sensing mechanism discussed in detail. Therefore, the investigated Pd-Pt/SiC/PSi structure may be a highly sensitive and selective hydrogen gas sensing electrode for deployment in extreme environment applications.

Keywords: RF Sputtering, silicon carbide, porous silicon, hydrogen gas sensor

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Authors: Andy Sand, Naama Massad – Ivanir, Nadav Nitzan, Elisa Valderrama, Alfred Wegenberger, Koranit Shlosman, Rotem Shemesh, Ester Segal

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Microbial spoilage of food products is of great concern in the food industry due to the direct impact on the shelf life of foods and the risk of foodborne illness. Therefore, food packaging may serve as a crucial contribution to keep the food fresh and suitable for consumption. Active packaging solutions that have the ability to inhibit the development of microorganism in food products attract a lot of interest, and many efforts have been made to engineer and assimilate such solutions on various food products. NanoPack is an EU-funded international project aiming to develop state-of-the-art antimicrobial packaging systems for perishable foods. The project is based on natural essential oils which possess significant antimicrobial activity against many bacteria, yeasts and molds. The essential oils are encapsulated in natural aluminosilicate clays, halloysite nanotubes (HNT's), that serves as a carrier for the volatile essential oils and enable their incorporation into polymer films. During the course of the project, several polyethylene films with diverse essential oils combinations were designed based on the characteristics of their target food products. The antimicrobial activity of the produced films was examined in vitro on a broad spectrum of microorganisms including gram-positive and gram-negative bacteria, aerobic and anaerobic bacteria, yeasts and molds. The films that showed promising in vitro results were successfully assimilated on in vivo active packaging of several food products such as cheese, bread, fruits and raw meat. The results of the in vivo analyses showed significant inhibition of the microbial spoilage, indicating the strong contribution of the NanoPack packaging solutions on the extension of shelf life and reduction of food waste caused by early spoilage throughout the supply chain.

Keywords: food safety, food packaging, essential oils, nanotechnology

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Authors: Serap Gedikli, Hakan Çakmak, M. Buğra Güldiken, Duygu Yalnızoğlu

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Air, which is necessary for living things to survive; while it carries some useful substances in it, it can also carry foreign particles of different sizes that may be harmful to the health. All airborne organic substances of biological origin, including bacteria, fungi, fungal spores, viruses, pollen, and their components, are called "bioaerosols". Nowadays, everyone spends most of their time in closed areas such as home, workplace, school, etc. Although it is known that outdoor air pollution affects health, it is not known that indoor air pollution has harmful effects in terms of health. In this study, indoor air microbial load and SARS-CoV-2 virus cleaning efficiency of Froumann brand air cleaners were studied. This work in 300 m³, 600 m³, and 1000 m³ completely closed areas without any air circulation with Froumann N80, N90, and N100 air-cleaning devices. Analyzes were performed for both areas at 60 minutes before and after the device was operated using a particle measuring device (Particles Plus 7302) and an air sampler (Mas-100 ECO). The measurements were taken by placing the test equipment 1.5-2 m away from the air cleaner. At the same time, the efficiency of the HEPA filter was evaluated by taking samples from the air outlet point of the HEPA filter using the air sampling device (Mas-100 ECO) after the device was started. Nutrient agar and malt agar are used as total mesophilic bacteria and total fungi. The number of colony-forming units per m³ (cfu/m³) was calculated by counting colonies in Petri dishes after incubation for 48 hours at 37°C for bacteria and 72 hours at 30°C for fungi. The change in the number of colonies and the decrease in the microbial load was calculated as a percentage value. SARS-CoV-2 activity analysis studies were carried out by İnönü University Microbiology Department in accordance with the World Health Organization regulations. Finally, the HEPA filter in the devices used was taken and kept under a certain temperature and humidity, and the change in the microbial load on it was monitored over a 6-month period. At the end of the studies, a 91%-94% reduction was determined in the total mesophilic bacteria count of Frouman brand N80, N90, and N100 model air cleaners. A decrease of 94%-96% was detected in the total number of yeast/molds. HEPA filter efficiency was evaluated, and at the end of the analysis, 98% of the bacterial load and approximately 100% of yeast/mold load at the HEPA filter air outlet point were decreased. According to the SARS- CoV-2 analysis results, when the device is operating at the medium airflow level 3, it can filter virus-carrying aerosols by 99%. As a result, it was determined that the Froumann model air cleaner was effective in controlling and reducing the microbial load in the indoor air.

Keywords: HEPA filter, indoor air quality, microbial load, SARS-CoV-2

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Authors: J. Tomilla Ajayi, Werner E. Van Zyl

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This work presents an overview of the reaction of 2, 4-diferrocenyl-1, 3-dithiadiphosphetane-2, 4-disulfide (Ferrocenyl Lawesson’s reagent) with water to produce the non-symmetric, ferocenyl dithiophosphonic acid respectively in high yields. These acids were readily deprotonated by anhydrous Ammonia to yield the corresponding ammonium salt NH4S2PFcOH. These were complex to Ni (II) in molar ratio 1:1 and 1:2. The resulting complex from the reaction formed same compound with different isomers (Cis and Trans) and also compound with multimetallic coordination. Quality X-ray crystals were formed from THF/Ether. The compounds were characterized by 1H, 31P NMR, and FTIR. Bulk purity were confirmed by either ESI-MS or elemental analysis and The XRD images were obtained using single crystal X-ray crystallographic studies. The electrochemical investigation of the Compounds were carried out using cyclic voltammetry.

Keywords: ferrocenyl, dithiophosphonate, isomer, coordination

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Authors: José Maria, Vânia Laranjo, Luís Abrunhosa, António Inês

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The occurrence of mycotoxigenic moulds such as Aspergillus, Penicillium and Fusarium in food and feed has an important impact on public health, by the appearance of acute and chronic mycotoxicoses in humans and animals, which is more severe in the developing countries due to lack of food security, poverty and malnutrition. This mould contamination also constitutes a major economic problem due the lost of crop production. A great variety of filamentous fungi is able to produce highly toxic secondary metabolites known as mycotoxins. Most of the mycotoxins are carcinogenic, mutagenic, neurotoxic and immunosuppressive, being ochratoxin A (OTA) one of the most important. OTA is toxic to animals and humans, mainly due to its nephrotoxic properties. Several approaches have been developed for decontamination of mycotoxins in foods, such as, prevention of contamination, biodegradation of mycotoxins-containing food and feed with microorganisms or enzymes and inhibition or absorption of mycotoxin content of consumed food into the digestive tract. Some group of Gram-positive bacteria named lactic acid bacteria (LAB) are able to release some molecules that can influence the mould growth, improving the shelf life of many fermented products and reducing health risks due to exposure to mycotoxins. Some LAB are capable of mycotoxin detoxification. Recently our group was the first to describe the ability of LAB strains to biodegrade OTA, more specifically, Pediococcus parvulus strains isolated from Douro wines. The pathway of this biodegradation was identified previously in other microorganisms. OTA can be degraded through the hydrolysis of the amide bond that links the L-β-phenylalanine molecule to the ochratoxin alpha (OTα) a non toxic compound. It is known that some peptidases from different origins can mediate the hydrolysis reaction like, carboxypeptidase A an enzyme from the bovine pancreas, a commercial lipase and several commercial proteases. So, we wanted to have a better understanding of this OTA degradation process when LAB are involved and identify which molecules where present in this process. For achieving our aim we used some bioinformatics tools (BLAST, CLUSTALX2, CLC Sequence Viewer 7, Finch TV). We also designed specific primers and realized gene specific PCR. The template DNA used came from LAB strains samples of our previous work, and other DNA LAB strains isolated from elderberry fruit, silage, milk and sausages. Through the employment of bioinformatics tools it was possible to identify several proteins belonging to the carboxypeptidase family that participate in the process of OTA degradation, such as serine type D-Ala-D-Ala carboxypeptidase and membrane carboxypeptidase. In conclusions, this work has identified carboxypeptidase proteins being one of the molecules present in the OTA degradation process when LAB are involved.

Keywords: carboxypeptidase, lactic acid bacteria, mycotoxins, ochratoxin a.

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