Search results for: OBD-plus microbial consortia

Authors: Sharda Nara, Bansi Dhar Malhotra

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Investigation of the vitamins as an electron transfer mediator could significantly assist in merging the area of tissue engineering and electronics required for the implantable therapeutic devices. The present study report that the molecules of folic acid released by Providencia rettgeri via fermentation route under the anoxic condition of the microbial fuel cell (MFC) exhibit characteristic electrochemical and optical properties, as indicated by absorption spectroscopy, photoluminescence (PL), and cyclic voltammetry studies. The absorption spectroscopy has depicted an absorption peak at 263 nm with a small bulge around 293 nm on day two of bacterial culture, whereas an additional peak was observed at 365 nm on the twentieth day. Furthermore, the PL spectra has indicated that the maximum emission occurred at various wavelengths 420, 425, 440, and 445 nm when excited by 310, 325, 350, and 365 nm. The change of emission spectra with varying excitation wavelength might be indicating the presence of tunable optical bands in the folic acid molecules co-related with the redox activity of the molecules. The results of cyclic voltammetry studies revealed that the oxidation and reduction occurred at 0.25V and 0.12V, respectively, indicating the electrochemical behavior of the folic acid. This could be inferred that the released folic acid molecules in a MFC might undergo inter as well as intra molecular electron transfer forming different intermediate states while transferring electrons to the electrode surface. Synchronization of electrochemical and optical properties of folic acid molecules could be potentially promising for the designing of electroactive scaffold and biocompatible conductive surface for the applications of tissue engineering and biofuel cells, respectively.

Keywords: biofuel cell, electroactivity, folic acid, tissue engineering

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Authors: Esma Hande Alıcı, Gülnur Arabacı

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Antioxidants are compounds or systems that can safely interact with free radicals and terminate the chain reaction before vital molecules are damaged. The anti-oxidative effectiveness of these compounds depends on their chemical characteristics and physical location within a food (proximity to membrane phospholipids, emulsion interfaces, or in the aqueous phase). Antioxidants (e.g., flavonoids, phenolic acids, tannins, vitamin C, vitamin E) have diverse biological properties, such as antiinflammatory, anti-carcinogenic and anti-atherosclerotic effects, reduce the incidence of coronary diseases and contribute to the maintenance of gut health by the modulation of the gut microbial balance. Plants are excellent sources of antioxidants especially with their high content of phenolic compounds. Raphanus raphanistrum L., the wild radish, is a flowering plant in the family Brassicaceae. It grows in Asia and Mediterranean region. It has been introduced into most parts of the world. It spreads rapidly, and is often found growing on roadsides or in other places where the ground has been disturbed. It is an edible plant, in Turkey its fresh aerial parts are mostly consumed as a salad with olive oil and lemon juice after boiled. The leaves of the plant are also used as anti-rheumatic in traditional medicine. In this study, we determined the antioxidant capacity of two different solvent fractions (methanol and ethyl acetate) obtained from Raphanus raphanistrum L. plant leaves. Antioxidant capacity of the plant was introduced by using three different methods: DPPH radical scavenging activity, CUPRAC (Cupric Ion Reducing Antioxidant Capacity) activity and Reducing power activity.

Keywords: antioxidant activity, antioxidant capacity, Raphanis raphanistrum L., wild radish

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Authors: Natcha Ruamyat, Nichakorn Khondee

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The potential of an alkaliphilic bacteria isolated from soil in Thailand to utilized agro-industrial and agricultural wastes for the production of biosurfactants was evaluated in this study. Among five isolates, Pseudomonas mendocina NK41 used soapstock as substrate showing a high biosurfactant concentration of 7.10 g/L, oil displacement of 97.8 %, and surface tension reduction to 29.45 mN/m. Various agricultural residues were applied as mixed substrates with soapstock to enhance the synthesis of biosurfactants. The production of biosurfactant and bacterial growth was found to be the highest with coconut oil cake as compared to Sacha inchi shell, coconut kernel cake, and durian shell. The biodegradability of agro-industrial wastes was better than agricultural wastes, which allowed higher bacterial growth. The pretreatment of coconut oil cake by combined alkaline and hydrothermal method increased the production of biosurfactant from 12.69 g/L to 13.82 g/L. The higher microbial accessibility was improved by the swelling of the alkali-hydrothermal pretreated coconut oil cake, which enhanced its porosity and surface area. The pretreated coconut oil cake was reused twice in the repeated batch production, showing higher biosurfactant concentration up to 16.94 g/L from the second cycle. These results demonstrated the capability of using lignocellulosic wastes from agricultural and agro-industrial activities to produce a highly valuable biosurfactant. High biosurfactant yield with low-cost substrate reveals its potential towards further commercialization of biosurfactant on large-scale production.

Keywords: alkaliphilic bacteria, agricultural/agro-industrial wastes, biosurfactant, combined alkaline-hydrothermal pretreatment

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Authors: Farideh Tabatabei-Yazdi, Fereshteh Falah, Alireza Vasiee

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Nowadays, production of functional foods has become very essential. Inulin is one of the most functional hydrocolloid compounds used in such products. In the present study, the production of a synbiotic yogurt containing 1, 2.5, and 5% (w/v) inulin has been investigated. The yogurt was fermented with Lactobacillus brevis PML1 derived from Tarkhineh, an Iranian cereal-dairy fermented food. Furthermore, the physicochemical properties, antioxidant activity, sensory attributes, and microbial viability properties were investigated on the 0th, 7th, and 14th days of storage after fermentation. The viable cells of L. brevis PML1 reached 108 CFU/g, and the product resisted to simulated digestive juices. Moreover, the synbiotic yogurt impressively increased the production of antimicrobial compounds and had the most profound antimicrobial effect on S. typhimurium. The physiochemical properties were in the normal range, and the fat content of the synbiotic yogurt was reduced remarkably. The antioxidant capacity of the fermented yogurt was significantly increased (p<0:05), which was equal to those of DPPH (69:18±1:00%) and BHA (89:16±2:00%). The viability of L. brevis PML1 was increased during storage. Sensory analysis showed that there were significant differences in terms of the impressive parameters between the samples and the control (p<0:05). Addition of 2.5% inulin not only improved the physical properties but also retained the viability of the probiotic after 14 days of storage, in addition to the viability of L. brevis with a viability count above 6 log CFU/g in the yogurt. Therefore, a novel synbiotic product containing L. brevis PML1, which can exert the desired properties, can be used as a suitable carrier for the delivery of the probiotic strain, exerting its beneficial health effects.

Keywords: functional food, lactobacillus brevis, symbiotic yogurt, physiochemical properties

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Authors: Syed Makhdoom Hussain, Muhammad Afzal, Farhat Jabeen, Arshad Javid, Tasneem Hameed

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A feeding trial was conducted with Cirrhinus mrigala fingerlings to study the effects of microbial phytase with graded levels (0, 500, 1000, 1500, and 2000 FTUkg-1) by sunflower meal based diet on growth performance and nutrient digestibility. The chromic oxide was added as an indigestible marker in the diets. Three replicate groups of 15 fish (Average wt 5.98 g fish-1) were fed once a day and feces were collected twice daily. The results of present study showed improved growth and feed performance of Cirrhinus mrigala fingerlings in response to phytase supplementation. Maximum growth performance was obtained by the fish fed on test diet-III having 1000 FTU kg-1 phytase level. Similarly, nutrient digestibility was also significantly increased (p<0.05) by phytase supplementation. Digestibility coefficients for sunflower meal based diet increased 15.76%, 17.70%, and 12.70% for crude protein, crude fat and apparent gross energy as compared to the reference diet, respectively at 1000 FTU kg-1 level. Again, maximum response of nutrient digestibility was recorded at the phytase level of 1000 FTU kg-1 diet. It was concluded that the phytase supplementation to sunflower meal based diet at 1000 FTU kg-1 level is optimum to release adequate chelated nutrients for maximum growth performance of C. mrigala fingerlings. Our results also suggested that phytase supplementation to sunflower meal based diet can help in the development of sustainable aquaculture by reducing the feed cost and nutrient discharge through feces in the aquatic ecosystem.

Keywords: sunflower meal, Cirrhinus mrigala, growth, nutrient digestibility, phytase

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Authors: Majid Javanmard

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Edible mushrooms are those agricultural products which contain high quantity of protein and can have special role in human diet. So search for methods to increase their shelf life is important. One of these strategies can be use of active packaging for absorb the ethylene which has been studied in present study. In this study, initially, production of impregnating zeolite with potassium permanganate has been studied with zeolite clinoptiolite available in iran. After that, these ethylene scavengers were placed in the package of edible mushrooms and then transferred to the refrigerator with temperature 4c for a period of 20 days. Each 5 days, several experiments accomplished on edible mushrooms such as weight loss, moisture content, color, texture, bacterial experiments and sensory evaluation. After production of impregnating zeolite with potassium permanganate (with a concentration of %2.5, %5, %7.5, %10 and %12.5) by zeolite type clinoptiolite (with mesh 35 and 60), samples have been analyzed with gas chromatography and titration with sodium oxalate. The results showed that zeolite by concentration of %5, %7.5 and %10 potassium permanganate and mesh 60 have a higher efficiency. Results from the experiments on edible mushrooms proved that impregnated zeolite with potassium permanganate have a meaningful influence in prevent the weight loss, decrease of moisture content and L-value, increase of a-value and overall color change (ΔE) and decrease of firmness texture of mushrooms. In addition, these absorbents can influence on decrease microbial load (mesophilic bacteria) rather than control. Generally, concluded that the impregnated zeolite with 10% permanganate potassium has a high efficiency on increase the shelf life of fresh edible mushrooms.

Keywords: active packaging, ethylene scavenger, zeolite clinoptiolite, permanganate potassium, shelf life

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Authors: Mubashir Hussain, Mu Lv, Xiaohan Dong, Zhiyang Li, Bin Liu, Nongyue He

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Detection and classification of microbes have a vast range of applications in biomedical engineering especially in detection, characterization, and quantification of bacterial contaminants. For identification of pathogens, different techniques are emerging in the field of biomedical engineering. Latest technology uses light scattering, capable of identifying different pathogens without any need for biochemical processing. Bacterial Pathogens Identification System (BPIS) which uses a laser beam, passes through the sample and light scatters off. An assembly of photodetectors surrounded by the sample at different angles to detect the scattering of light. The algorithm of the system consists of two parts: (a) Library files, and (b) Comparator. Library files contain data of known species of bacterial microbes in the form of binned plots, while comparator compares data of unknown sample with library files. Using collected data of unknown bacterial species, highest voltage values stored in the form of peaks and arranged in 3D histograms to find the frequency of occurrence. Resulting data compared with library files of known bacterial species. If sample data matching with any library file of known bacterial species, sample identified as a matched microbe. An experiment performed to identify three different bacteria particles: Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli. By applying algorithm using library files of given samples, results were compromising. This system is potentially applicable to several biomedical areas, especially those related to cell morphology.

Keywords: microbial identification, laser scattering, peak identification, binned plots classification

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Authors: Benselama Amel, Ounane S. Mohamed, Bekki Abdelkader

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A collection of 20 isolates from fresh Nodules of the legume plant Lablab purpureus was isolated. These isolates have been authenticated by seedling inoculation grown in jars containing sand. The results obtained after two months of culture have revealed that the 20 isolates (100% of the isolates) are able to nodulate their host plants. The results obtained were analyzed statistically by ANOVA using the software statistica and had shown that the effect of the inoculation has significantly improved all the growth parameters (the height of the plant and the dry weight of the aerial parts and roots, and the number of nodules). We have evaluated the tolerance of all strains of the collection to the major stress factors as the salinity, pH and extreme temperature. The osmotolerance reached a concentration up to 1710mm of NaCl. The strains were also able to grow on a wide range of pH, ranging from 4.5 to 9.5, and temperature, between 4°C and 40°C. Also, we tested the effect of the acidity, aluminum and ferric deficit on the Lablab-rhizobia symbiosis. Lablab purpureus has not been affected by the presence of high concentrations of aluminum. On the other hand, iron deficiency has caused a net decrease in the dry biomass of the aerial part. The results of all the phenotypic characters have been treated by the statistical Minitab software, the numerical analysis had shown that these bacterial strains are divided into two distinct groups at a level of similarity of 86 %. The SDS-PAGE was carried out to determine the profile of the total protein of the strains. The coefficients of similarity of polypeptide bands between the isolates and strains reference (Bradyrhizobium, Mesorizobium sp.) confirm that our strain belongs to the groups of rhizobia.

Keywords: SDS-PAGE, rhizobia, symbiosis, phenotypic characterization, Lablab purpureus

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Authors: Futo Asano, Yusuke Yatsushiro, Hirokuni Miyamoto, Hiroaki Kodama

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A thermophile-fermented compost is produced using small fishes, crabs, and shrimps under a high temperature (approximately 75℃) by fermentation-associated self-heating. This compost has been used as a feed additive for pigs and hens in Japan, and the fecundity of this livestock is enhanced. Firmicutes is a dominant phylum in the microbial composition of the compost. We first reported that improvement of female larval fitness of Hercules beetle can be achieved by amendment of this compost to the humus. When the 90-d-old larvae were reared for subsequent 72 days in the humus with this compost, the growth of female larvae was significantly enhanced when compared with the growth of female larvae in the humus without the compost. In contrast, the growth of male larvae in the compost-free humus was the same as the larvae grow in the compost-amended humus. The bacterial composition of the feces of larvae was determined at 0 days and 46 days after transfer to the humus with or without the compost. The most dominant bacterium in the feces was Xylanimonas. Interestingly, the growth improvement of female larvae was associated with an increased abundance of Mollicutes in the fecal samples. These results indicate that the compost act as a probiotic material for enhancing the female larvae growth by supporting Mollicutes. Here, we tried to isolate Mollicutes from the contents of the midgut and hindgut of the 3rd instar female larvae of the Hercules beetle. These gut contents were spread onto a selective agar medium for Mollicutes (PPLO agar broth, BD Difco, NJ, USA). Although we isolated none of the Mollicutes until now, several bacteria that are closely related to Xylanimonas and Luteimicrobium were isolated. These isolates have xylanase and glucanase (CMCase) activities. We show the gut bacterial profiles of larvae and discuss how the fitness of female larvae of the Hercules beetle is improved by the compost.

Keywords: compost, beetle, mollicutes, woody biomass

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Authors: Oluyemi O. Awolusi, Abimbola M. Enitan, Sheena Kumari, Faizal Bux

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Nitrification is essential to biological processes designed to remove ammonia and/or total nitrogen. It removes the excess nitrogenous compound in wastewater which could be very toxic to the aquatic fauna or cause a serious imbalance of such aquatic ecosystem. Efficient nitrification is linked to an in-depth knowledge of the structure and dynamics of the nitrifying community structure within the wastewater treatment systems. In this study, molecular technique was employed for characterizing the microbial structure of activated sludge [ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB)] in a municipal wastewater treatment with intention of linking it to the plant efficiency. PCR-based phylogenetic analysis was also carried out for. The average operating and environmental parameters, as well as specific nitrification rate of a plant, was investigated during the study. During the investigation, the average temperature was 23±1.5oC. Other operational parameters such as mixed liquor suspended solids and chemical oxygen demand inversely correlated with ammonia removal. The dissolved oxygen level in the plant was constantly lower than the optimum (between 0.24 and 1.267 mg/l) during this study. The plant was treating wastewater with the influent ammonia concentration of 31.69 and 24.47 mg/l. The influent flow rates (ML/day) was 96.81 during the period. The dominant nitrifiers include: Nitrosomonas spp. Nitrobacter spp. and Nitrospira spp. The AOB had a correlation with nitrification efficiency and temperature. This study shows that the specific ammonia oxidizing rate and the specific nitrate formation rates can serve as a good indicator of the plant overall nitrification performance.

Keywords: Ammonia monooxygenase α-subunit gene, amoA, ammonia-oxidizing bacteria, AOB, nitrite-oxidizing bacteria, NOB, specific nitrification rate

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Authors: Nuzhat Sultana M. B.

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Children are high-risk population in terms of food born illnesses. Food safety and security are the most important aspect of the success of midday meal programmes. Improper holding temperatures, cross-contamination and poor personal hygiene of food handlers are the main causes for the prevalence of pathogenic microbes in the food servicing areas. Two hundred and fifty preschool children in the age of 3 to 6 years from urban and rural anganwadies (pre school center) of Beed district were selected. Nutritional status of preschool children were assessed by anthropometrical and clinical measurement. The study assessed the food safety and security with the help of personal hygiene and other safety measures maintained by the food personnel working for midday meal programme, supplying mid meals to children in govt. anganwadies (pre school center). The hygiene level, sanitary condition and microbial quality of food and water, pathological health examination of food handlers were assessed with the help of checklist. A questionnaire was designed to evaluate knowledge, attitude, and practices of food handlers. Results of the study show that the nutritional and health status of rural and urban preschool children was very poor. Many of the food handlers were not aware of general knowledge and hygiene practices to be followed during food preparation areas. An intervention programme of education and importing training at workplaces has shown a positive impact on the outcome of safety and security practices and safe, hygienic practices of food handlers at workplace.

Keywords: food, health, preschool children, safety, security

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Authors: Asma Ansari, Afsheen Aman, Shah Ali Ul Qader

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In the past few years, numerous concerns have been raised against increased bacterial resistance towards effective drugs and become a debated issue all over the world. With the emergence of drug resistant pathogens, the interaction of natural antimicrobial compounds and antibacterial nanoparticles has emerged as a potential candidate for combating infectious diseases. Microbial diversity in the biome provides an opportunity to screen new species which are capable of producing large number of antimicrobial compounds. Among these antimicrobial compounds, bacteriocins are highly specific and efficient antagonists. A combination of bacteriocin along with nanoparticles could prove to be more potent due to broadened antibacterial spectrum with possibly lower doses. In the current study, silver nanoparticles were synthesized through biological reduction using various isolated bacterial, fungal and yeast strains. Spectroscopy and scanning electron microscopy (SEM) was performed for the confirmation of nanoparticles. Bacteriocin was characterized and purified to homogeneity through gel permeation chromatography. The estimated molecular weight of bacteriocin was 10 kDa. Amino acid analysis and N-terminal sequencing revealed the novelty of the protein. Then antibacterial potential of silver nanoparticles and broad inhibitory spectrum bacteriocin was determined through agar well diffusion assay. These synthesized bacteriocin-Nanoparticles exhibit a good potential for clinical applications as compared to bacteriocin alone. This combination of bacteriocin with nanoparticles will be used as a new sort of biocide in the field of nano-proteomics. The advancement of nanoparticles-mediated drug delivery system will open a new age for rapid eradication of pathogens from biological systems.

Keywords: BAC-IB17, multidrug resistance, purification, silver nanoparticles

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Authors: Amir Saber Gharamaleki, Beitollah Alipour, Zeinab Faghfoori, Ahmad YariKhosroushahi

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Colorectal cancer (CRC) is one of the most prevalent cancers and intestinal microbial community plays an important role in colorectal tumorigenesis. Probiotics have recently been assessed as effective anti-proliferative agents and thus this study was performed to examine whether CRC undergo apoptosis by treating with isolated Iranian native dairy yeast, Kluyveromyces marxianus YAS, secretion metabolites. The cytotoxicity assessments on cells (HT-29, Caco-2) were accomplished through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as well as qualitative DAPI (4',6-diamidino-2-phenylindole staining) and quantitative (flow cytometry assessments) evaluations of apoptosis. To evaluate the main mechanism of apoptosis, Real time PCR method was applied. Kluyveromyces marxianus YAS secretions (IC50) showed significant cytotoxicity against HT-29 and Caco-2 cancer cell lines (66.57 % and 66.34 % apoptosis) similar to 5-Fluorouracil (5-FU) while apoptosis only was developed in 27.57 % of KDR normal cells. The prophylactic effects of Kluyveromyces marxianus (PTCC 5195), as a reference yeast, was not similar to Kluyveromyces marxianus YAS indicating strain dependency of bioactivities on CRC disease prevention. Based on real time PCR results, the main cytotoxicity is related to apoptosis phenomenon and the core related mechanism is depended on the overexpression of BAX, CASP 9, CASP 8 and CASP 3 inducing apoptosis genes. However, several investigations should be conducted to precisely determine the effective compounds to be used as anticancer therapeutics in the future.

Keywords: anticancer, anti-proliferative, apoptosis, cytotoxicity, yeast

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Authors: Yael Shai, Dror L. Angel, Dror Zurel, Peleg Astrahan, Maxim Rubin-Blum, Eyal Rahav

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The high demand for oil and its by-products is symptomatic of the 21st century and occasionally leads to oil spills and pollution of coastal waters. Marine oil pollution may originate from a variety of sources -urban runoff, tanker cleaning, drilling activities, and oil spills. These events may release large amounts of highly toxic polycyclic aromatic hydrocarbons (PAHs) and other pollutants to coastal water, thereby threatening local marine life. Here, we investigated the effects of crude oil on the temporal dynamics of phytoplankton and heterotrophic bacteria in Israeli coastal waters. To this end, we added crude oil (500 µm thick layer, with and without additional nutrients; NO₃ and PO₄) to mesocosms (1m³ bags) containing oligotrophic surface coastal water collected near Haifa during summer and winter. Changes in phytoplankton biomass, activity and diversity were monitored daily for 5-6 days. Our results demonstrate that crude oil addition resulted in a pronounced decrease in phytoplankton biomass and production rates, while heterotrophic bacterial production increased significantly. Importantly, a few days post addition we found that the oil-degrading bacteria, Oleibacter sp. and Oleispira sp. appeared in the mesocosms and that the addition of nutrients (along with the crude oil) further increased this trend. This suggests that oil-degrading bacteria may be NO₃ and PO₄ limited in Israeli coastal waters. The results of this study should enable us to establish improved science-based environmental policy with respect to handling crude oil pollution in this region.

Keywords: heterotrophic bacteria, nutrients, mesocosm, oil pollution, oligotrophic, phytoplankton

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Authors: Zoran Herceg, Tomislava Vukušić, Anet Režek Jambrak, Višnja Stulić

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Today one of the greatest challenges are directed to the safety of food supply. If food pathogens are ingested they can cause human illnesses. Because of that new technologies that are effective in microbial reduction are developing to be used in food industries. One of such technology is cold gas phase plasma. Salmonella enterica was studied as one of the pathogenes that can be found in food. The aim of this work was to examine the inactivation rate and stress response of plasma treated cells of Salmonella enterica inoculated in apple juice. After the treatment cellular leakage, phenotypic changes in plasma treated cells-biofilm formation and degree of recovery were conducted. Sample volume was inoculated with 5 mL of pure culture of Salmonella enterica and 15 mL of apple juice. Statgraphics Centurion software (StatPoint Technologies, Inc., VA, USA) was used for experimental design and statistical analyses. Treatment time (1, 3, 5 min) and gas flow (40, 60, 80 L/min) were changed. Complete inactivation and 0 % of recovery after the 48 h was observed at these experimental treatments: 3 min; 40 L/min, 3 min; 80 L/min, 5 min; 40 L/min. Biofilm reduction was observed at all treated samples. Also, there was an increase in cellular leakage with a longer plasma treatment. Although there were a significant reduction and 0 % of recovery after the plasma treatments further investigation of the method is needed to clarify whether there are sensorial, physical and chemical changes in juices after the plasma treatment. Acknowledgments: The authors would like to acknowledge the support by Croatian Science Foundation and research project 'Application of electrical discharge plasma for the preservation of liquid foods'.

Keywords: salmonella enterica serotype typhimurium lt21, gas-phase plasma treatment, inactivation, stress response

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Authors: Christiana Ene Ogwuche, Joseph Amupitan, George Ndukwe, Rachael Ayo

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Maesobotrya barteri (Baill), belonging to the family Euphorbiaceae, is a medicinal plant growing widely in tropical Africa. The Aerial plant parts of Maesobotrya barteri (Baill) were collected fresh from Orokam, Ogbadibo local Government of Benue State, Nigeria in July 2013. Taxonomical identification was done by Mallam Musa Abdullahi at the Herbarium unit of Biological Sciences Department, ABU, Zaria, Nigeria. Pulverized aerial parts of Maesobotrya barteri (960g) was exhaustively extracted successively using petroleum ether, chloroform, ethyl acetate and methanol and concentrated in the rotary evaporator at 40°C. The Petroleum ether extract had the second highest activity against test microbes from preliminary crude microbial screenings. The Petroleum ether extract was subjected to phytochemical studies, antimicrobial analysis and column chromatography (CC). The column chromatography yielded fraction PE, which was further purified using preparative thin layer chromatography to give PE1. The structure of the isolated compound was established using 1-D NMR and 2-D NMR spectroscopic analysis and by direct comparison with data reported in literature was confirmed to be a mixture, an isomer of Betulinic acid and Oleanolic acid, both with the molecular weight (C₃₀H₄₈O₃). The bioactivity of this compound was carried out using some clinical pathogens and the activity compared with standard drugs, and this was found to be comparable with the standard drug.

Keywords: Maesobotrya barteri, medicinal plant, bioactivity, petroleum spirit extract, butellinic acid, oleanilic acid

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Authors: Christina Killian, Katie Wall, Séamus Fanning, Guerrino Macori

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Deep-level metagenomics offers a useful technical approach to explore the three dynamic One Health axes: healthcare, domiciliary and veterinary. There is currently limited understanding of the composition of complex biofilms, natural abundance of AMR genes and gene transfer occurrence in these ecological niches. By using a newly established small-scale complex biofilm model, COMBAT has the potential to provide new information on microbial diversity, antimicrobial resistance (AMR)-encoding gene abundance, and their transfer in complex biofilms of importance to these three One Health axes. Shotgun metagenomics has been used to sample the genomes of all microbes comprising the complex communities found in each biofilm source. A comparative analysis between untreated and biocide-treated biofilms is described. The basic steps include the purification of genomic DNA, followed by library preparation, sequencing, and finally, data analysis. The use of long-read sequencing facilitates the completion of metagenome-assembled genomes (MAG). Samples were sequenced using a PromethION platform, and following quality checks, binning methods, and bespoke bioinformatics pipelines, we describe the recovery of individual MAGs to identify mobile gene elements (MGE) and the corresponding AMR genotypes that map to these structures. High-throughput sequencing strategies have been deployed to characterize these communities. Accurately defining the profiles of these niches is an essential step towards elucidating the impact of the microbiota on each niche biofilm environment and their evolution.

Keywords: COMBAT, biofilm, metagenomics, high-throughput sequencing

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Authors: Bilge Kaan Tekelioglu, Mehmet Celik, Mahmut Ali Gokce, Ladine Celik, Yusuf Uzun

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A male mature barn owl (Tyto alba) was brought to Cukurova University Ceyhan Veterinary Medicine Faculty at the beginning of January 2017. The bird was found at a local state elementary school’s garden where had been terribly damaged by metal wires. On the clinical examination, the animal was in shock and atonic position at arrival and seems to have feather problems and severe injuries. The ears, eyes, claws and wounded areas were checked and no signs of viral, microbial or ecto-parasitic infection were observed. The bird has been declared by U.S. wild life Office as endangered species. At first, the owl was kept in silent, warm and darkened cabinet against shock and warmed fluid replacement was started by % 5 dextrose solution per orally. On the second day, we started per oral forced feeding with chicken flesh meat dipped into the dextrose solution. On the third day, the bird was continued to be fed with fresh meat. At the fourth day, the owl was started to be fed with chicks during the next 3 days died by natural means which has been supplied by a local breeder. At the first 3 days 1 chick per day and the following days 2 chicks per day has been given per orally. The tenth day we started flying exercises in a small and non-windowed room safely. The saved owl was kept in this room for 10 more days. Finally, the owl was released at the habitation where it had been found injured. This study has one more time proved that, if you save one, you can save more. Wild life is in danger all over the world. Every living creature has right and deserves a chance to live.

Keywords: wild life, barn owl, Tyto alba, rescue, life support, feeding

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Authors: Fahad Aljasass, Hamza Abu-Tarboush, Salah Aleid, Siddig Hamad

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The effects of different high hydrostatic pressure treatments on the shelf life of camel’s milk were studied. Treatments at 300 to 350 MPa for 5 minutes at 40°C reduced microbial contamination to levels that prolonged the shelf life of refrigerated (3° C) milk up to 28 days. The treatment resulted in a decrease in the proteolytic activity of the milk. The content of proteolytic enzymes in the untreated milk sample was 4.23 µM/ml. This content decreased significantly to 3.61 µM/ml when the sample was treated at 250 MPa. Treatment at 300 MPa decreased the content to 3.90 which was not significantly different from the content of the untreated sample. The content of the sample treated at 350 MPa dropped to 2.98 µM/ml which was significantly lower than the contents of all other treated and untreated samples. High pressure treatment caused a slight but statistically significant increase in the pH of camel’s milk. The pH of the untreated sample was 6.63, which increased significantly to 6.70, in the samples treated at 250 and 350 MPa, but insignificantly in the sample treated at 300 MPa. High pressure treatment resulted in some degree of milk fat oxidation. The thiobarbituric acid (TBA) value of the untreated sample was 0.86 mg malonaldehyde/kg milk. This value remained unchanged in the sample treated at 250 MPa, but then it increased significantly to 1.25 and 1.33 mg/kg in the samples treated at 300 and 350 MPa, respectively. High pressure treatment caused a small increase in the greenness (a* value) of camel’s milk. The value of a* was reduced from -1.17 for the untreated sample to -1.26, -1.21 and -1.30 for the samples treated at 250, 300 and 350 MPa, respectively. Δa* at the 250 MPa treatment was -0.09, which then decreased to -0.04 at the 300 MPa treatment to increase again to -0.13 at the 350 MPa treatment. The yellowness (b* value) of camel’s milk increased significantly as a result of high pressure treatment. The b* value of the untreated sample was 1.40, this value increased to 2.73, 2.31 and 2.18 after treatments at 250, 300 and 350 MPa, respectively. The Δb* value was +1.33 at the treatment 250 MPa, decreased to +0.91 at 300 MPa and further to +0.78 at 350 MPa. The pressure treatment caused slight effect on color, slight decrease in protease activity and a slight increase in the oxidation products of lipids.

Keywords: high hydrostatic pressure, camel’s milk, mesophilic aerobic bacteria, clotting, protease

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Authors: Alok Kumar, Hari Ram, Lebin Thomas, Ved Pal Singh

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Organic solvent tolerant amylases and proteases of microbial origin are in great demand for their application in transglycosylation of water-insoluble flavanoids and in peptide synthesizing reaction in organic media. Most of the amylases and proteases are unstable in presence of organic solvent. In the present work two different bacterial strains M-11 and VP-07 were isolated from the soil sample of a dairy farm in Delhi, India, for the efficient production of extracellular amylase and protease through their screening on starch agar (SA) and skimmed milk agar (SMA) plates, respectively. Both the strains (M-11 and VP-07) were identified based on morphological, biochemical and 16S rRNA gene sequencing methods. After analysis through Ez-Taxon software, the strains M-11 and VP-07 were found to have maximum pairwise similarity of 98.63% and 100% with Bacillus subtilis subsp. inaquosorum BGSC 3A28 and Bacillus anthracis ATCC 14578 and were therefore identified as Bacillus sp. UKS1 and Bacillus sp. UKS2, respectively. Time course study of enzyme activity and bacterial growth has shown that both strains exhibited typical sigmoid growth behavior and maximum production of amylase (180 U/ml) and protease (78 U/ml) by these strains (UKS1 and UKS2) was commenced during stationary phase of growth at 24 and 20 h, respectively. Thereafter, both amylase and protease were tested for their tolerance towards organic solvents and were found to be active as well stable in p-xylene (130% and 115%), chloroform (110% and 112%), isooctane (119% and 107%), benzene (121% and 104%), n-hexane (116% and 103%) and toluene (112% and 101%, respectively). Owing to such properties, these enzymes can be exploited for their potential application in industries for organic synthesis.

Keywords: amylase, enzyme activity, industrial applications, organic solvent tolerant, protease

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Authors: Munish Garg, Vinni Ahuja

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In the past few years, the high consumption of soft drinks has attracted negative attention world-wide due to its possible adverse effects, leading the health conscious people to find alternative nutraceutical or herbal health drinks. In the present study, a nutraceutical soft drink was developed utilizing some easily available and well known traditional herbs having nutritional potential. The key ingredients were selected as bael, amla, lemon juice, ashwagandha and poppy seeds based on their household routine use in the summer with proven refreshing, cooling and energetic feeling since ages. After several trials made, the final composition of nutraceutical summer soft drink was selected as most suitable combination based on the taste, physicochemical, microbial and organoleptic point of view. The physicochemical analysis of the prepared drink found to contain optimum level of titratable acidity, total soluble solids and pH which were in accordance of the commercial recommendations. There were no bacterial colonies found in the product therefore found within limits. During the nine point’s hedonic scale sensory evaluation, the drink was strongly liked for colour, taste, flavour and texture. The formulation was found to contain flavonoids (80mg/100ml), phenolics (103mg/100ml), vitamin C (250mg/100ml) and has antioxidant potential (75.52%) apart from providing several other essential vitamins, minerals and healthy components. The developed nutraceutical drink provides an economical and feasible option for the consumers with very good taste combined with potential health benefits. The present drink is potentially capable to replace the synthetic soft drinks available in the market.

Keywords: herbal drink, summer drink, nutraceuticals, soft drink

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Authors: Carolina Guatemala, Josefina Barrera

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A large number of soils highly polluted with recalcitrant hydrocarbons and the limitation of the current bioremediation methods continue being the drawback for an efficient recuperation of these under safe conditions. In this regard, soil washing by degradable surfactants is an alternative option knowing the capacity of surfactants to desorb oily organic compounds. The aim of this study was the establishment of the washing conditions of a soil polluted with diesel, using a nonionic surfactant. A soil polluted with diesel was used. This was collected near to a polluted railway station zone. The soil was dried at room temperature and sieved to a mesh size 10 for its physicochemical and biological characterization. Washing of the polluted soil was performed with surfactant solutions in a 1:5 ratio (5g of soil per 25 mL of the surfactant solution). This was carried out at 28±1 °C and 150 rpm for 72 hours. The factors tested were the Tween 80 surfactant concentration (1, 2, 5 and 10%) and the treatment time. Residual diesel concentration was determined every 24 h. The soil was of a sandy loam texture with a low concentration of organic matter (3.68%) and conductivity (0.016 dS.m- 1). The soil had a pH of 7.63 which was slightly alkaline and a Total Petroleum Hydrocarbon content (TPH) of 11,600 ± 1058.38 mg/kg. The high TPH content could explain the low microbial count of 1.1105 determined as UFC per gram of dried soil. Within the range of the surfactant concentration tested for washing the polluted soil under study, TPH removal increased proportionally with the surfactant concentration. 5080.8 ± 422.2 ppm (43.8 ± 3.64 %) was the maximal concentration of TPH removed after 72 h of contact with surfactant pollution at 10%. Despite the high percentage of hydrocarbons removed, it is assumed that a higher concentration of these could be removed if the washing process is extended or is carried out by stages. Soil washing through the use of surfactants as a desorbing agent was found to be a viable and effective technology for the rapid recovery of soils highly polluted with recalcitrant hydrocarbons.

Keywords: diesel, hydrocarbons, soil washing, tween 80

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Authors: María D. Groppa, Andrea Trentini, Myriam Zawoznik, Roxana Bigi, Carlos Nadra, Patricia L. Marconi

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In the present work, a remediation bioprocess based on the use of a local isolate of the microalgae Chlorella vulgaris immobilized in alginate beads is proposed. This process was shown to be effective for the reduction of several chemical and microbial contaminants present in Cildáñez stream, a water course that is part of the Matanza-Riachuelo Basin (Buenos Aires, Argentina). The bioprocess, involving the culture of the microalga in autotrophic conditions in a stirred-tank bioreactor supplied with a marine propeller for 6 days, allowed a significant reduction of Escherichia coli and total coliform numbers (over 95%), as well as of ammoniacal nitrogen (96%), nitrates (86%), nitrites (98%), and total phosphorus (53%) contents. Pb content was also significantly diminished after the bioprocess (95%). Standardized cytotoxicity tests using Allium cepa seeds and Cildáñez water pre- and post-remediation were also performed. Germination rate and mitotic index of onion seeds imbibed in Cildáñez water subjected to the bioprocess was similar to that observed in seeds imbibed in distilled water and significantly superior to that registered when untreated Cildáñez water was used for imbibition. Our results demonstrate the potential of this simple and cost-effective technology to remove urban-water contaminants, offering as an additional advantage the possibility of an easy biomass recovery, which may become a source of alternative energy.

Keywords: bioreactor, bioremediation, Chlorella vulgaris, Matanza-Riachuelo Basin, microalgae

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Authors: Q. Rzina, M. Lahrouni, S. Rida, N. Saadaoui, Y. Almossaid, K. Oufdou, K. Fares

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Many organic solid wastes are produced in the world. Poultry manure (PM), municipal organic wastes (MOW) and sugar beet lime sludge (LS) are produced in large quantities in Morocco. The co-composting of these organic wastes was investigated. The recycling and the valorization of such wastes is environmentally and economically beneficial especially for PM which is known source of bacterial pathogens. The aerobic biodegradation process was carried out by using three windrows of variable compositions: C1 prepared without LS (only MOW were composted with PM), C2 prepared from MOW plus PM and10% LS; and the last one C3 from MOW plus PM and 20% LS. The main process physico-chemical parameters (temperature, pH, humidity and C/N) and microbiological populations (mesophilic and thermophilic flora, total coliform, fecal coliform, Streptococci, Staphylococcus aureus and mesophilic fungi) were monitored over three months to ascertain the compost maturity and to ensure the compost hygienic aspect. The final products were characterized by their relatively high organic matter content, and low C/N ratio of 10.6-10.9. The organic matter degradation was reached approximately 59% for C2 and C3. In addition, the monitoring of the microbial population showed that the produced composts are mature and hygienic. The agronomic valorization of the final composts was tested on radish plant with tree level of composts and poultry manure without composting. The primary results of field trial showed a growth of radish plant biomass and root development without any phytotoxicity detected which reflects the quality of the composts produced. As for poultry manure it allowed to have a better results than other composts because of its readily available nitrogen.

Keywords: compost, municipal organic wastes, poultry manure, radish crop, sugar beet lime sludge

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Authors: Patrick Mueller, Alan Martin, Justin Stockwell, Robert Goldblatt

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Elevated concentrations of inorganic nitrogen (N) compounds (nitrate, nitrite, and ammonia) are a ubiquitous feature to mine-influenced drainages due to the leaching of blasting residues and use of cyanide in the milling of gold ores. For many mines, the management of N is a focus for environmental protection, therefore understanding the factors controlling the speciation and behavior of N is central to effective decision making. In this paper, the passive attenuation of ammonia and nitrite is described for three northern water bodies (two lakes and a tailings pond) influenced by mining activities. In two of the water bodies, inorganic N compounds originate from explosives residues in mine water and waste rock. The third water body is a decommissioned tailings impoundment, with N compounds largely originating from the breakdown of cyanide compounds used in the processing of gold ores. Empirical observations from water quality monitoring indicate nitrification (the oxidation of ammonia to nitrate) occurs in all three waterbodies, where enrichment of nitrate occurs commensurately with ammonia depletion. The N species conversions in these systems occurred more rapidly than chemical oxidation kinetics permit, indicating that microbial mediated conversion was occurring, despite the cool water temperatures. While nitrification of ammonia and nitrite to nitrate was the primary process, in all three waterbodies nitrite was consistently present at approximately 0.5 to 2.0 % of total N, even following ammonia depletion. The persistence of trace amounts of nitrite under these conditions suggests the co-occurrence denitrification processes in the water column and/or underlying substrates. The implications for N management in mine waters are discussed.

Keywords: explosives, mining, nitrification, water

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Authors: Mudawi M. Nour

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Background: Now a days, the multidisciplinary scientific research conception in the field of nanotechnology has witnessed development with regard to the numerous applications and synthesis of nanomaterials. Objective: The current investigation has been conducted with the main focus on the green synthesis of silver nanoparticles from the leaves of Salvadora persica and its antibacterial activity against MDR pathogens E. coli and S. aureus. Methodology: Silver nanoparticles (AgNPs) were prepared after addition of aqueous extract of Salvadora persica leaves. The UV-Vis spectrophotometer, Transmission Electron Microscopy (TEM), zeta potential and Scanning Electron Microscopy (SEM) were employed to detect the particle size and morphology, besides Fourier transform infra-red spectrometer (FTIR) analysis was performed to determine the capping and stabilizing agents in the extract. Antibacterial assay for the biogenic AgNPs was conducted against E. coli and S. aureus. Results: Color change of the mixture from yellow to dark brown is the first indication to AgNPs formation. Furthermore, 420 nm was the peak value for UV-Vis spectroscopy absorption of the mixture. Besides, TEM and SEM micrographs showed wide variability in the diameter of smaller NPs aggregated together with spherical shapes, and zeta sizer showed about 153.3 nm as an average size of nanoparticles. Microbial suppression was noticed for the tested microorganisms. Furthermore, with the help of FTIR analysis, the biomolecules that act as capping and stabilizing agents of AgNPs are proteins and phenols present in the plant extract. Conclusion: Salvadora persica leaves extract act as a reducing and stabilizing agent for the synthesis of AgNPs, keeping its ability to suppress the MDR pathogen.

Keywords: green synthesis, FTIR, MDR pathogen, salvadora persica

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Authors: Angeliki Vassila, Dimitrios Kalogeropoulos, Rania Rawashdeh, Nigel Hall, Najiha Rahman, Mark Fabian, Suresh Thulasidharan, Hossain Parwez

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Introduction: Necrotising scleritis is a severe form of scleritis and poses a significant threat to vision. It can manifest in various systemic autoimmune disorders, systemic vasculitis, or as a consequence of microbial infections. The objective of this study is to present a case of necrotizing scleritis associated with scleroderma, which was further complicated by a secondary Staphylococcus epidermidis infection. Methods: This is a retrospective analysis that examines the medical records of a patient who was hospitalised in the Eye Unit at University Hospital Southampton. Results: A 78-year-old woman presented at the eye casualty department of our unit with a two-week history of progressively worsening pain in her left eye. She received a diagnosis of necrotising scleritis and was admitted to the hospital for further treatment. It was decided to commence a three-day course of intravenous methylprednisolone followed by a tapering regimen of oral steroids. Additionally, a conjunctival swab was taken, and two days later, it revealed the presence of S. epidermidis, indicating a potential secondary infection. Given this finding, she was also prescribed topical (Ofloxacin 0.3% - four times daily) and oral (Ciprofloxacin 750mg – twice daily) antibiotics. The inflammation and symptoms gradually improved, leading to the patient being scheduled for a scleral graft and applying an amniotic membrane to cover the area of scleral thinning. Conclusions: Rheumatoid arthritis and granulomatosis with polyangiitis are the most commonly identifiable systemic diseases associated with necrotising scleritis. Although association with scleroderma is extremely rare, early identification and treatment are necessary to prevent scleritis-related complications.

Keywords: scleritis, necrotizing scleritis, scleroderma, autoimmune disease

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Authors: Zainab Bibi, Afsheen Aman, Shah Ali Ul Qader

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Endo-β-1,4-xylanases [EC 3.2.1.8] are one of the major groups of enzymes that are involved in degradation process of xylan and have several applications in food, textile and paper processing industries. Due to broad utility of endo-β-1,4-xylanase, researchers are focusing to increase the productivity of this hydrolase from various microbial species. Harsh industrial condition, faster reaction rate and efficient hydrolysis of xylan with low risk of contamination are critical requirements of industry that can be fulfilled by synthesizing the enzyme with efficient properties. In the current study, a newly isolated thermophile Geobacillus stearothermophilus KIBGE-IB29 was used in order to attain the maximum production of endo-1,4-β-xylanase. Bacterial culture was isolated from soil, collected around the blast furnace site of a steel processing mill, Karachi. Optimization of various nutritional and physical factors resulted the maximum synthesis of endo-1,4-β-xylanase from a thermophile. High production yield was achieved at 60°C and pH-6.0 after 24 hours of incubation period. Various nitrogen sources viz. peptone, yeast extract and meat extract improved the enzyme synthesis with 0.5%, 0.2% and 0.1% optimum concentrations. Dipotassium hydrogen phosphate (0.25%), potassium dihydrogen phosphate (0.05%), ammonium sulfate (0.05%) and calcium chloride (0.01%) were noticed as valuable salts to improve the production of enzyme. The thermophilic nature of isolate, with its broad pH stability profile and reduced fermentation time indicates its importance for effective xylan saccharification and for large scale production of endo-1,4-β-xylanase.

Keywords: geobacillus, optimization, production, xylanase

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Authors: Hadjer Didouh, Izzaddine Sameut Bouhaik, Mohammed Hadj Meliani

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This research delves into the intricate dynamics of biofilm adhesion on surfaces, particularly focusing on the widely employed X52 surface in oil and gas industry pipelines. Biofilms, characterized by microorganisms within a self-produced matrix, pose significant challenges due to their detrimental impact on surfaces. Our study integrates advanced molecular techniques and cutting-edge microscopy, such as scanning electron microscopy (SEM), to identify microbial communities and visually assess biofilm adhesion. Simultaneously, we concentrate on the X52 surface, utilizing impedance spectroscopy and potentiodynamic polarization to gather electrochemical responses under various conditions. In conjunction with the broader investigation, we propose a novel approach to mitigate biofilm-induced corrosion challenges. This involves environmentally friendly inhibitors derived from plants, offering a sustainable alternative to conventional chemical treatments. Our inquiry screens and selects inhibitors based on their efficacy in hindering biofilm formation and reducing corrosion rates on the X52 surface. This study contributes valuable insights into the interplay between electrochemical processes and biofilm attachment on the X52 surface. Furthermore, the outcomes of this research have broader implications for the oil and gas industry, where biofilm-related corrosion is a persistent concern. The exploration of eco-friendly inhibitors not only holds promise for corrosion control but also aligns with environmental considerations and sustainability goals. The comprehensive nature of this research aims to enhance our understanding of biofilm dynamics, provide effective strategies for corrosion mitigation, and contribute to sustainable practices in pipeline management within the oil and gas sector.

Keywords: bio-corrosion, biofilm, attachment, X52, metal/bacteria interface

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Authors: Pampita Chakraborty, Sukumar Mukherjee

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The study was done to determine the microbiological profile and changing pattern of the pathogens causing UTI in the ICU patients. All the patients admitted to the ICU with urinary catheter insertion for more than 48hours were included in the study. Urine samples were collected in a sterile container with aseptic precaution using disposable syringe and was processed as per standards. Antimicrobial susceptibility test was done by Disc Diffusion method as per CLSI guidelines. A total of 100 urine samples were collected from ICU patients, out of which 30% showed significant bacterial growth and 7% showed growth of candida spp. Prevalence of UTI was more in female (73%) than male (27.%). Gram-negative bacilli 26(86.67%) were more common in our study followed by gram-positive cocci 4(13.33%). The most common uropathogens isolated were Escherichia coli 14 (46.67%), followed by Klebsiella spp 7(23.33%), Staphylococcus aureus 4(13.33%), Acinetobacter spp 3(10%), Enterococcus faecalis 1(3.33%) and Pseudomonas aeruginosa 1(3.33%). Most of the Gram-negative bacilli were sensitive to amikacin (80%) and nitrofurantoin (80%), where as all gram-positive organisms were sensitive to Vancomycin. A large number ESBL producers were also observed in this study. The study finding showed that E.coli is the predominant pathogen and has increasing resistance pattern to the commonly used antibiotics. The study proposes that the adherence to antibiotic policy is the key ingredients for successful outcome in ICU patients and also emphasizes that repeated evaluation of microbial characteristics and continuous surveillance of resistant bacteria is required for selection of appropriate antibiotic therapy.

Keywords: antimicrobial sensitivity, intensive care unit, nosocomial infection, urinary tract infection

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