Search results for: Escherichia coli O157:H7

Authors: Aida Kalantari, Boyang Ji, Tao Chen, Ivan Mijakovic

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3-hydroxypropanoic acid (3-HP) is one of the most important biomass-derivable platform chemicals that can be converted into a number of industrially important compounds. There have been several attempts at production of 3-HP from renewable sources in cell factories, focusing mainly on Escherichia coli, Klebsiella pneumoniae, and Saccharomyces cerevisiae. Despite the significant progress made in this field, commercially exploitable large-scale production of 3-HP in microbial strains has still not been achieved. In this study, we investigated the potential of Bacillus subtilis to be used as a microbial platform for bioconversion of glycerol into 3-HP. Our recombinant B. subtilis strains overexpress the two-step heterologous pathway containing glycerol dehydratase and aldehyde dehydrogenase from various backgrounds. The recombinant strains harboring the codon-optimized synthetic pathway from K. pneumoniae produced low levels of 3-HP. Since the enzymes in the heterologous pathway are sensitive to oxygen, we had to perform our experiments in micro-aerobic conditions. Under these conditions, the cell produces lactate in order to regenerate NAD+, and we found the lactate production to be in competition with the production of 3-HP. Therefore, based on the in silico predictions, we knocked out the glycerol kinase (glpk), which in combination with growth on glucose, resulted in improving the 3-HP titer to 1 g/L and the removal of lactate. Cultivation of the same strain in an enriched medium improved the 3-HP titer up to 7.6 g/L. Our findings provide the first report of successful introduction of the biosynthetic pathway for conversion of glycerol into 3-HP in B. subtilis.

Keywords: bacillus subtilis, glycerol, 3-hydroxypropanoic acid, metabolic engineering

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Authors: Adrian Ionut Nicoara, Denisa Ionela Ene, Alina Maria Holban, Cristina Busuioc

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At present, the development of materials with biomedical applications is a domain of interest that will produce a full series of benefits in engineering and medicine. In this sense, it is required to use a natural material, and this paper is focused on the development of a composite material based on bacterial cellulose – hydroxyapatite and silver nanoparticles with applications in hard tissue. Bacterial cellulose own features like biocompatibility, non-toxicity character and flexibility. Moreover, the bacterial cellulose can be conjugated with different forms of active silver to possess antimicrobial activity. Hydroxyapatite is well known that can mimic at a significant level the activity of the initial bone. The material was synthesized by using an ultrasound probe and finally characterized by several methods. Thereby, the morphological properties were analyzed by using Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). Because the synthesized material has medical application in restore the tissue and to fight against microbial invasion, the samples were tested from the biological point of view by evaluating the biodegradability in phosphate-buffered saline (PBS) and simulated body fluid (SBF) and moreover the antimicrobial effect was performed on Gram-positive bacterium Staphylococcus aureus, Gram-negative bacterium Escherichia coli, and fungi Candida albicans. The results reveal that the obtained material has specific characteristics for bone regeneration.

Keywords: bacterial cellulose, biomaterials, hydroxyapatite, scaffolds materials

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Authors: Rafael Estrada, Cristian Ruiz Rueda

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Carbapenems are last-resort antibiotics used in clinical settings to treat antibiotic-resistant bacterial infections. Thus, the emergence and spread of resistance to carbapenems is a major public health concern. Here, we have studied a carbapenem-resistant Aeromonas veronii strain previously isolated from a water sample from Sam Simeon Creek (Hearst San Simeon State Park, CA). Analysis of this isolate using disk-diffusion, CarbaNP, eCIM and mCIM assays revealed that it was resistant to amoxicillin-clavulanic acid and all carbapenems tested and that this isolate produced a potentially novel carbapenemase of the Metallo-β-lactamase family. Whole genome sequencing analysis revealed that this A. veronii isolate carries a novel variant of the blacₚₕₐ class β-carbapenemase gene that was closely related to the blacₚₕₐ₇ gene of Aeromonas jandaei. This isolate also carried a novel variant of the blaₒₓₐ class D carbapenemase gene that was most closely related to the blaₒₓₐ-₉₁₂ gene found in other Aeromonas veronii isolates. Finally, we also identified a novel class C β-lactamase gene moderately related to the blaFₒₓ-₁₇ gene of Providencia stuartii and other blaFₒₓ variants identified in Klebsiella pneumoniae, Escherichia coli and other Enterobacteriaceae. Overall, our findings reveal that environmental isolates are an important reservoir of multiple carbapenemases and other β-lactamases of clinical significance.

Keywords: β-lactamases, carbapenem, antibiotic-resistant, aeromonas veronii

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Authors: K. P. Ganesh, T. Gomathi, L. Arul Pragasan

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Degradation of lake water quality is one of the serious environmental threats for the last few decades, particularly, the lakes situated in and around urban and industrial areas. The present study aimed to analyze the physicochemical and biological parameters, and metal elements to determine the water quality of Krishnampathi, Ukkadam, Kurichi, Sulur and Singanallur Lakes. Of the 23 physicochemical parameters analyzed in the five lakes, except TDS, Chloride and Total hardness values all the 20 parameters were found within the prescribed limit as recommended by World Health Organization (WHO) and Bureau of Indian Standards (BIS). In case of biological parameter, both Total Coliform and Fecal Coliform bacteria (Escherichia coli) were identified. This indicates the contamination of lakes by fecal matter, and warns of potential of disease causing by viruses, bacteria and other organisms. Among the twelve metal elements (Al, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Mo, Cd and Pb) determined by inductively coupled plasma-mass spectroscopy, except Cd (for all lakes), and Pb (for Ukkadam, Kurichi, Sulur & Singanallur), all the elements were found above the prescribed limits of BIS. The results of the present study revealed that all the five major lakes of Coimbatore were contaminated. It is recommended that proper implementation of the new wetland waste management system and monitoring of water quality be of the urgent need to sustain the water bodies for future generations.

Keywords: heavy metals, inductively coupled plasma-mass spectroscopy, physicochemical and biological parameters, water quality

Procedia PDF Downloads 157

Authors: Alvine Sandrine Ndinchout, D. P. Chattopadhyay, Moundipa Fewou Paul, Nyegue Maximilienne Ascension, Varinder Kaur, Sukhraj Kaur, B. H. Patel

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Dacryodes macrophylla is a species of the Burseraceae family that is widespread in Cameroon, Equatorial Guinea, and Gabon. The only part of D. macrophylla known to use is the pulp contained in the fruit. This very juicy pulp is consumed directly and used in making juices. During consumption, these fruit leaves a dark blackish colour on fingers and garment. This observation means that D. macrophylla fruits must be a good source of natural dye with probably good fastness properties on textile materials. But D. macrophylla has not yet been investigated with reference as a potential source of natural dye to our best knowledge. Natural dye has been extracted using water as solvent by soxhlet extraction method. The extracted color was characterized by spectroscopic studies like UV/Visible and further tested for antimicrobial activity against gram-negative (Vibrio cholerae, Escherichia coli, Salmonella enterica serotype Typhi, Shigella flexneri) and gram-positive (Listeria monocytogenes, Staphylococcus aureus) bacteria. It was observed that the water extract of D. macrophylla showed antimicrobial activities against S. enterica. The results of fastness properties of the dyed fabrics were fair to good. Taken together, these results indicate that D. macrophylla can be used as natural dye not only in textile but also in other domains like food coloring.

Keywords: antimicrobial activity, natural dye, silk, wash fastness, wool

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Authors: Hayet El Kolli, Hocine Laouer

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Bunium in the world comprises about 50 to 100 species, mostly distributed in: Algeria, Italy, Pakistan, Iran, and South Africa. Bunium species have several uses like: Bunium persicum which is commonly used as antispasmodic, carminative, anti-obesity and lactogage. This plant have been widely used as an additive in food stuff such as in bread cooking, rice and yoghurt for its carminative, anti-dyspepsia and antispasmodic effect. The B. paucifolium oil has a wide spectrum of action against moulds, yeast and bacteria. The chemical compositions of Bunium incrassatum and Bunium alpinum essential oils were carry out by GC and GC/MS. Therefore, antibacterial activity of two oils was investigated by disk diffusion method against Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 1331, Staphylococcus aureus ATCC 25923, Klebsiella pneumoniae ATCC 700603, Bacillus cereus ATCC 10876, Enterococcus faecalis ATCC 49452, Lysteria monocytogenes ATCC 15313, Citrobacter freundii ATCC 8090, Proteus mirabilis ATCC 35659. A moderate antibacterial activity was found. In conclusion, it is found that essential oils of the two species are rich in sesquiterpens and other oxygenated compounds. These compounds have been reported to show bactericidal activity and the presence of phenolic compounds makes them useful antioxidants so that results confirm some ethnopharmacologique applications of these two oils of Bunium.

Keywords: Bunium alpinum, Bunium incrassatum, apiaceae, essential oil, sesquiterpens, phenols, antibacterial, antioxidant activities

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Authors: Elmineh Tsegahun Gedif

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Zinc oxide nanoparticles (ZnO NPs) have attracted huge attention due to catalytic, optical, photonic, and antibacterial activity. Zinc oxide nanoparticles were successfully synthesized via a fast, non-toxic, cost-effective, and eco-friendly method by biologically reducing Zn(NO3)2.6H2O solution with Neem (Azadirachta indica) leaf extract under optimum conditions (pH = 9). The presence of active flavonoids, phenolic groups, alkaloids, terpenoids, and tannins, which were in the biomass of the Neem leaf extract before and after reduction, was identified using qualitative screening methods (observing the color changes) and FT-IR Spectroscopy. The formation of ZnO NPs was visually indicated by the color changes from colorless to light yellow color. Biosynthesized nanoparticles were also characterized by UV-visible, FT-IR, and XRD spectroscopies. The reduction process was simple and convenient to handle and was monitored by UV-visible spectroscopy that showed surface plasmon resonance (SPR) of the ZnO NPs at 321 nm. This result clearly revealed the formation of ZnO NPs. X-ray diffraction was used to investigate the crystal structure. The average particle size of ZnO powder and around 20 nm using the line width of the plane, and the refraction peak using Scherrer’s equation. The synthesized zinc oxide nanoparticles were evaluated for antimicrobial activities against Gram-positive and Gram-negative bacteria. Zinc nanoparticles exhibited the maximum zone of inhibition against Escherichia coli (15 mm), while the least activity was seen against Staphylococcus aureus.

Keywords: antimicrobial activity, azadirachta indica, green synthesis, ZnO NPs

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Authors: Muhammad Arshad, Ghulam Hussain Bhatti, Abdul Qayyum

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Zinc-doped silica oxide nanoparticles having size 7.93nm were synthesized by the deposition precipitation method by using different solvents (acetonitrile, n-hexane, isoamylalchol). Biological potential such as antibacterial activities against Bacillussubtilusand Escherichia coli, and antifungal activities against Candida parapsilosis and Aspergilusniger were also investigated by Disc diffusion method. Different characterizations techniques including Fournier Transmission Infrared Spectroscopy (FT-IR), X-ray diffraction (XRD), Scanning Electron Microscope (SEM), Transmission Electron Microscope (TEM), Thermo-gravimeteric Analysis (TGA), Atomic forced microscopy (AFM), and Dynamic Light Scattering (DLS) were used. FT-IR characterization confirmed the presence of metal oxide bond (SiO2) while XRD showed the hexagonal structure. SEM and TEM characterization showed the morphology of nanoparticles. AFM study showed good particle size distribution as depicted by a histogram. DLS study showed the gradual decease in the size of nanoparticles from 24.86nm to 13.24 nm. Highest antibacterial activities revealed by acetonitrile solvents (6%and 4.5%) followed by isoamylalchol (3% and 2.4%) while n-hexane solvent showed the lowest activity (2%and 1%) respectively. Higher antifungal activities exhibited by n-hexane (0.34 % and 0.43%) followed by isoamylalchol (0.27% and 0.19%) solvent while acetonitrile (0.21% and 0.17%) showed least activity respectively. Statistical analysis by using one-way ANOVA also indicated the significant results of both biological activities.

Keywords: nanoparticles, precipitation methods, antibacterial, antifungal, characterizations

Procedia PDF Downloads 180

Authors: Justin Ntokamunda Kadima, Christian Ahadi Irenge, Patient Birindwa Mulashe, Félicien Mushagalusa Kasali, Patient Wimba

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Background: Bacterial strains carrying multidrug resistance traits are gaining ground worldwide, especially in countries with limited resources. This study aimed to evaluate the spreading of multidrug-resistant bacteria strains in Bukavu city hospitals in the Democratic Republic of Congo. Methods: We analyzed 758 antibiogram data recorded in files of patients consulted between January 2016 and December 2017 at three reference hospitals selected as sentinel sites, namely the Panzi General Reference Hospital (HGP), BIO -PHARM hospital (HBP), and Saint Luc Clinic (CSL). Results: Of 758 isolates tested, the laboratories identified 12 bacterial strains in 712 isolates, of which 223 (29.42%) presented MDR profile, including Escherichia coli (11.48%), Klebsiella pneumonia (6.07%), Enterobacter (5.8%), Staphylococcus aureus and coagulase-negative Staphylococci (1.58%), Proteus mirabilis (1.85%), Salmonella enterica (1.19%), Pseudomonas aeruginosa (0.53%), Streptococcus pneumonia (0.4%)), Citrobacter (0.13%), Neisseria gonorrhea (0.13%), Enterococcus faecalis (0.13%), and Morganella morganii (0.13%). Infected patients were significantly more adults (73.1% vs. 21.5%) compared to children and mainly women (63.7% vs. 30.9%; p = 0.001). Conclusion: The observed expansion requires that hospital therapeutic committees set up an effective clinical management system and define the right combinations of antibiotics.

Keywords: multidrug resistance, bacteria, antibiogram, Bukavu

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Authors: Khaled Khleifat

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This study detects the bacterial species that responsible for UTI in both diabetic patients and non-diabetic patients, Jordan. 116 urine samples were investigated in order to determine UTI-causing bacteria. These samples distributed unequally between diabetic male (12) and diabetic female (25) and also non-diabetic male (13) and non-diabetic female (66). The results represent that E.coli is responsible for UTI in both diabetic and non-diabetic patients (15.5% and 29.3% respectively) with large proportion (44.8%). This study showed that not all bacterial species that isolated from the non-diabetic sample could be isolated from diabetic samples. E. coli (15.5%), P. aeruginosa (4.3%), K. pneumonia (1.7%), P. mirabilis (2.6%), S. marcescens (0.9%), S. aureus (1.7%), S. pyogenes (1.7%), E. faecalis (0.9%), S. epidermidis (1.7%) and S. saprophyticus (0.9%). But E. aerogenes, E. cloacae, C. freundii, A. baumannii and B. subtilis are five bacterial species that can’t isolate from all diabetic samples. This study shows that for the treatment of UTI in both diabetic and non-diabetic patients, Chloramphenicol (30 μg), Ciprofloxacin (5 μg) and Vancomycin (30 μg) are more favorable than other antibiotics. In the same time, Cephalothin (30μg) is not recommended.

Keywords: urinary tract infections, diabetes mellitus, bacterial species, infections

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Authors: Kirstin Burger, Paul Watts, Nicole Vorster

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Green chemistry focuses on synthesis which has a low negative impact on the environment. This research focuses on synthesizing novel compounds from an all-natural Eucalyptus citriodora oil. Eight novel plasticizer compounds are synthesized and optimized using flow chemistry technology. A precursor to one novel compound can be synthesized from the lauric acid present in coconut oil. Key parameters, such as catalyst screening and loading, reaction time, temperature, residence time using flow chemistry techniques is investigated. The compounds are characterised using GC-MS, FT-IR, 1H and 13C-NMR techniques, X-ray crystallography. The efficiency of the compounds is compared to two commercial plasticizers, i.e. Dibutyl phthalate and Eastman 168. Several PVC-plasticized film formulations are produced using the bio-based novel compounds. Tensile strength, stress at fracture and percentage elongation are tested. The property of having increasing plasticizer percentage in the film formulations is investigated, ranging from 3, 6, 9 and 12%. The diastereoisomers of each compound are separated and formulated into PVC films, and differences in tensile strength are measured. Leaching tests, flexibility, and change in glass transition temperatures for PVC-plasticized films is recorded. Research objective includes using these novel compounds as a green bio-plasticizer alternative in plastic products for infants. The inhibitory effect of the compounds on six pathogens effecting infants are studied, namely; Escherichia coli, Staphylococcus aureus, Shigella sonnei, Pseudomonas putida, Salmonella choleraesuis and Klebsiella oxytoca.

Keywords: bio-based compounds, plasticizer, tensile strength, microbiological inhibition , synthesis

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Authors: Abdul Walusansa

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In this paper, paper notes and coins were collected from general public in Kampala City where ready-to-eat food can be served, in order to survey for bacterial contamination. The total bacterial number and potentially pathogenic organisms loading on currency were tested. All isolated potential pathogens were also tested for antibiotic resistance against four most commonly prescribed antibiotics. 1. The bacterial counts on one hundred paper notes sample were ranging between 6～10918/cm cm-2,the median was 141/ cm-2, according to the data it was much higher than credit cards and Australian notes which were made of polymer. The bacterial counts on sixty coin samples were ranging between 2～380/cm-2, much less than paper notes. 2. Coliform (65.6%), E. coli (45.9%), S. aureus (41.7%), B. cereus (67.7%), Salmonella (19.8%) were isolated on one hundred paper notes. Coliform (22.4%), E. coli (5.2%), S. aureus (24.1%), B. cereus (34.5%), Salmonella (10.3%) were isolated from sixty coin samples. These results suggested a high rate of potential pathogens contamination of paper notes than coins. 3. Antibiotic resistances are commonly in most of the pathogens isolated on currency. Ampicillin resistance was found in 60%of Staphylococcus aureus isolated on currency, as well as 76.6% of E. coil and 40% of Salmonella. Erythromycin resistance was detected in 56.6% of S. aureus and in 80.0% of E. coli. All the pathogens isolated were sensitive to Norfloxacin, Salmonella and S. aureus also sensitive to Cefaclor. In this paper, we also studied the antimicrobial capability of metal coins, coins collected from different countries were tested for the ability to inhibit the growth of E. sakazakii, S. aureus, E. coli, L. monocytogenes and S. typhimurium. 1) E. sakazakii appeared very sensitive to metal coins, the second is S. aureus, but E. coli, L. monocytogenes and S. typhimurium are more resistant to these metal coin samples. 2) Coins made of Nickel-brass alloy and Copper-nickel alloy showed a better effect in anti-microbe than other metal coins, especially the ability to inhibited the growth of E. sakazakii and S. aureus, all the inhibition zones produced on nutrient agar are more than 20.6 mm. Aluminium-bronze alloy revealed weak anti-microbe activity to S. aureus and no effect to kill other pathogens. Coins made of stainless steel also can’t resist bacteria growth. 3) Surprisingly, one cent coins of USA which were made of 97.5% Zinc and 2.5% Cu showed a significant antimicrobial capability, the average inhibition zone of these five pathogens is 45.5 mm.

Keywords: antibiotic sensitivity, bacteria, currency, coins, parasites

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Authors: Saqib Ali, Man-Qun Wang

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The Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae), is a major pest of pines and it is also the key vector of the exotic pinewood nematode in China. In the present study, we cloned, expressed, and purified a chemosensory protein (CSP) in M. alternatus. We surveyed its expression in various developmental stages of male and female adult tissues and determined its binding affinities for different pine volatiles using a competitive binding fluorescence assay. A CSP known as CSP5 in M. alternatus was obtained from an antennal cDNA library and expressed in Escherichia coli. Quantitative reverse transcription polymerase chain reaction results indicated that the CSP5 gene was mainly expressed in male and female antennae. Competitive binding assays were performed to test the binding affinity of recombinant CSP5 to 13 odour molecules of pine volatiles. The results showed that CSP5 showed very strong binding abilities to myrcene, (+)-β-pinene, and (−)-isolongifolene, whereas the volatiles 2-methoxy-4-vinylphenol, p-cymene, and (+)-limonene oxide have relatively weak binding affinity at pH 5.0. Three volatiles myrcene, (+)-β-pinene, and (−)-isolongifolene may play crucial roles in CSP5 binding with ligands, but this needs further study for confirmation. The sensitivity of insect to host plant volatiles can effectively be used to control and monitor the population through mass trapping as part of integrated pest management programs.

Keywords: olfactory-specific protein, volatiles, competitive binding assay, expression characteristics, qPCR

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Authors: Sachin Latiyan, T. S. Sampath Kumar, Mukesh Doble

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Natural polymer based nanofibrous wound dressings have gained increased attention because of their high surface area, bioactivity, biodegradability and resemblance to extracellular matrix. Agarose (a natural polymer) have been used largely for angiogenesis, cartilage formation and wound healing applications. However, electrospinning of agarose is tedious thereby rendering limited studies on fabrication and evaluation of agarose based nanofibrous wound dressings. Thus, present study focuses on the fabrication of agarose (10% w/v)/ polyvinyl alcohol (12% w/v) based multifunctional nanofibrous scaffolds. Zinc citrate (1, 3 and 5% w/w of the polymer) was added as a potential antibacterial agent to combat wound infections. The fabricated scaffolds exhibit ~500% swelling (in phosphate buffer saline) with enhanced mechanical strength which is suitable for most of the wound healing applications. In vitro studies were found to reveal an increased migration and proliferation of L929 mouse fibroblasts with agarose blends w.r.t to the control. The fabricated dressings were found to be effective against both Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive) bacterial strains. Hence, a multifunctional (as provides effective swelling and mechanical support along with antibacterial property), natural product based, eco-friendly scaffold was successfully fabricated to serve as a potential wound dressing material.

Keywords: antibacterial dressings, benign solvent, nanofibrous agarose, biocompatibility, enhanced swelling and mechanical strength, biopolymeric dressings

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Authors: Solomon Gebremariam, Mulugeta Naizigi, Aregawi Haileselassie

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Background: Knowledge of the pattern of bacterial isolates and their antimicrobial susceptibility is crucial for guiding empirical treatment and infection prevention and control measures. Objective: The aim of this study was to analyze the pattern of bacterial isolates and their susceptibility patterns from various specimens. Methods: Retrospectively, a total of 1067 microbiological culture results that were isolated, characterized, and identified by standard microbiological methods and whose antibiotic susceptibility was determined using CLSI guidelines between 2017 and 2019 were retrieved and analyzed. Data were entered and analyzed using the Stata release 10.1 statistical package. Result: The positivity rate of culture was 26.04% (419/1609). The most common bacteria isolated were S. aureus 23.8% (94), E. coli 15.1% (60), Klebsiella pneumonia 14.1% (56), Pseudomonas aeruginosa 8.5% (34), and CONS 7.3% (29). S. aureus and CONS showed a high (58.1% - 96.2%) rate of resistance to most antibiotics tested. They were less resistant to Vancomycin which is 18.6% (13/70) and 11.8% (2/17), respectively. Similarly, the resistance of E. coli, Klebsella pneumonia, and Pseudomonas aeruginosa was high (69.4% - 100%) to most antibiotics. They were less resistant to Ciprofloxacilin, which is 41.1% (23/56), 19.2% (10/52), and 16.1% (5/31), respectively. Conclusion: This study has shown that there is a high rate of antibiotic resistance among bacterial isolates in this hospital. A combination of Vancomycin and Ciprofloxacin should be considered in the choice of antibiotics for empirical treatment of suspected infections due to S. aureus, CONS, E. coli, Klebsiella pneumonia, Pseudomonas such as in infections within hospital setup.

Keywords: antimicrobial, resistance, bacteria, hospital

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Authors: Sarita Puri, Tapan K. Chaudhuri

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Chaperonin GroEL is a tetradecameric Escherichia coli protein having identical subunits of 57 kDa. The elucidation of thermodynamic parameters related to stability for the native GroEL is not feasible as it undergoes irreversible unfolding because of its large size (800kDa) and multimeric nature. Nevertheless, it is important to determine the thermodynamic stability parameters for the highly stable GroEL protein as it helps in folding and holding of many substrate proteins during many cellular stresses. Properly folded monomers work as building-block for the formation of native tetradecameric GroEL. Spontaneous refolding behavior of monomeric GroEL makes it suitable for protein-denaturant interactions and thermodynamic stability based studies. The urea mediated unfolding is a three state process which means there is the formation of one intermediate state along with native and unfolded states. The heat mediated denaturation is a two-state process. The unfolding process is reversible as observed by the spontaneous refolding of denatured protein in both urea and head mediated refolding processes. Analysis of folding/unfolding data provides a measure of various thermodynamic stability parameters for the monomeric GroEL. The proposed mechanism of unfolding of monomeric GroEL is a three state process which involves formation of one stable intermediate having folded apical domain and unfolded equatorial, intermediate domains. Research in progress is to demonstrate the importance of specific residues in stability and oligomerization of GroEL protein. Several mutant versions of GroEL are under investigation to resolve the above mentioned issue.

Keywords: equilibrium unfolding, monomeric GroEl, spontaneous refolding, thermodynamic stability

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Authors: E. Bartkiene, M. Ruzauskas, V. Lele, P. Zavistanaviciute, J. Bernatoniene, V. Jakstas, L. Ivanauskas, D. Zadeike, D. Klupsaite, P. Viskelis, J. Bendoraitiene, V. Navikaite-Snipaitiene, G. Juodeikiene

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In this study, antimicrobial nutraceuticals; gummy candies (GC) from bovine colostrum (BC), essential oils (EOs), probiotic lactic acid bacteria (PLAB), and their combinations, were developed. For antimicrobial GC preparation, heteropolysaccharide (agar) was used. The antimicrobial properties of EOs (Eugenia caryophyllata, Thymus vulgaris, Citrus reticulata L., Citrus paradisi L.), BC, L. paracasei LUHS244, L. plantarum LUHS135, and their combinations against pathogenic bacteria strains (Streptococcus mutans, Enterococcus faecalis, Staphylococcus aureus, Salmonella enterica, Escherichia coli, Proteus mirabilis, and Pseudomonas aeruginosa) were evaluated. The highest antimicrobial properties by EO’s (Eugenia caryophyllata and Thymus vulgaris) were established. The optimal ingredients composition for antimicrobial GC preparation was established, which incorporate the BC fermented with L. paracasei LUHS244 in combination with Thymus vulgaris or Eugenia caryophyllata. These ingredients showed high inhibition properties of all tested pathogenic strains (except Pseudomonas aeruginosa). Antimicrobial GC formula consisting of thyme EO (up to 0.2%) and fermented BC (up to 3%), and for taste masking, mandarin or grapefruit EOs (up to 0.2%) was used. Developed GC high overall acceptability and antimicrobial properties, thus, antimicrobial GC could be a preferred form of nutraceuticals. This study was fulfilled with the support of the LSMU-KTU joint project.

Keywords: antimicrobial activity, bovine colostrum, essential oil, gummy candy, probiotic

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Authors: Anushaa A.

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In this work, silver nanoparticles (AgNP) were manufactured directly without harmful chemicals utilising methanol extract (SNLME) Solanum nigrume leaves. We are using nigrum leaf extract from Solanum, which converts silver nitrate to silver ions, for synthesization purposes. An examination of the AgNP produced was performed using ultraviolet (UV-VIS) spectroscopy, infrared spectroscopy (FTIR) transformed from Fourier and scanning electrons (SEM). Biological activity was also tested. UV-VIS has proven that biosynthesized AgNP exists (420-450 nm). The FTIR spectrum has been utilised to confirm the presence of different functional groups within the biomolecules, which are a nanoparticular capping agent and the spectroscopic and crystal nature of AgNP. The viability of the silver nanoparticles was evaluated using zeta potential calculations. Negative zeta potential of -33.4 mV demonstrated the stability of silver-nanoparticles. The morphology of AgNP was examined using a scanning electron microscope. Greenly generated AgNP showed significant anti-Staphylococcus aureus, Candida, and Escherichia coli action. The green AgNP demonstration indicated that the IC50 for the human teratocarcinoma cell line was 29.24 μg/ml during 24 hours of therapy (PA1 Ovarian cell line). The dose-dependent effects were reported in both antibacterial and cytotoxicity assays and as an effective agent. Finally, the findings of this research showed that silver nanoparticles generated might serve as a viable therapeutic agent to combat microorganisms killing and curing cancer.

Keywords: antimicrobial activity, PA1 ovarian cancer cell line, silver nanoparticles, Solanum nigrum

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Authors: Hafiza Javaria Ashraf, Xinghong Wang, Zhanghong Shi, Youming Hou

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Insect’s innate immunity depends on a variety of defense responses for the recognition of invading pathogens. Pathogen recognition involves particular proteins known as pattern recognition receptors (PRRs). These PRRs interact with pathogen-associated molecular patterns (PAMPs) present on the surface of pathogens to distinguish between self and non-self. C-type lectins (CTLs) belong to a superfamily of PPRs which involved in insect immunity and defense mechanism. Rhynchophorus ferrugineus Olivier is a devastating pest of Palm cultivations in China. Although studies on R. ferrugineus immune mechanism and host defense have conducted, however, the role of CTL in immune responses of R. ferrugineus remains elusive. Here, we report RfCTL, which is a secreted protein containing a single-CRD domain. The open reading frame (ORF) of CTL is 226 bp, which encodes a putative protein of 168 amino acids. Transcript expression analysis revealed that RfCTL highly expressed in immune-related tissues, i.e., hemolymph and fat body. The abundance of RfCTL in the gut and fat body dramatically increased upon Staphylococcus aureus and Escherichia coli bacterial challenges, suggesting a role in defense against gram-positive and gram-negative bacterial infection. Taken together, we inferred that RfCTL might be involved in the immune defense of R. ferrugineus and established a solid foundation for future studies on R. ferrugineus CTL domain proteins for better understanding of insect immunity.

Keywords: biological invasion, c-type lectin, insect immunity, Rhynchophorus ferrugineus Oliver

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Authors: Tejinder Pal Singh, Ravinder Kumar Malik, Gurpreet Kaur

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Adhesion is key factor for colonization of the gastrointestinal tract and the ability of probiotic strains to inhibit pathogens. Therefore, the adhesion ability is considered as a suitable biomarker for the selection of potential probiotic. In the present study, eight probiotic Lactobacillus reuteri strains were evaluated as viable, LiCl treated or heat-killed forms and compared with probiotic reference strains (L. reuteri ATCC55730). All strains investigated were able to adhere to Caco-2 cells. All probiotic L. reuteri strains tested were able to inhibit and displace (P < 0.05) the adhesion of Escherichia coli ATCC25922, Salmonella typhi NCDC113, Listeria monocytogenes ATCC53135 and Enterococcus faecalis NCDC115. The probiotic strain L. reuteri LR6 showed the strongest adhesion and pathogen inhibition ability among the eight L. reuteri strains tested. In addition, the abilities to inhibit and to displace adhered pathogens depended on both the probiotic and the pathogen strains tested suggesting the involvement of various mechanisms. The adhesion and antagonistic potential of the probiotic strains were significantly decreased upon exposure to 5M LiCl, showing that surface molecules, proteinaceous in nature, are involved. The heat-killed forms of the probiotic L. reuteri strains also inhibited the attachment of selected pathogens to Caco-2 cells. In conclusion, in vitro assays showed that L. reuteri strains, as viable or heat-killed forms, are adherent to Caco-2 cell line model and are highly antagonistic to selected pathogens in which surface molecules, proteinaceous molecules in particular, plays an important role.

Keywords: probiotics, Lactobacillus reuteri, adhesion, Caco-2 cells

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Authors: Sajjad Jafari, Reza Akbari

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Background and Aim: Chrysin, a flavonoid compound found in medicinal plants, honey, and propolis, has potential biological activities that make it an important perspective in the treatment of infectious and cancer diseases. The aim of this review study is to evaluate the biological activities of chrysin in the treatment of infectious and cancer diseases. Material and Methods: The present study is a review study that searched reputable scientific databases such as PubMed, Google Scholar, Scopus, and Web of Science from 2000 to 2023 using keywords such as antimicrobial, antifungal, chrysin, anticancer, antioxidants, and infectious diseases. The researchers examined 25 articles to determine the biological activities of chrysin. Results: Chrysin has high inhibitory or lethal activities on gram-positive and gram-negative bacteria, including Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Enterococcus faeces. It also has anti-biofilm effects and antifungal effects on strains such as Aspergillus niger and Candida albicans. Chrysin also has anticancer effects on various cancers, including colorectal cancer, pancreatic cancer, breast cancer, and MCF-7 cancer, which have been confirmed in vitro and in vivo. Conclusion: Chrysin has the potential as an important therapeutic option in the treatment of infectious and cancer diseases. Its high antimicrobial and anticancer activities, combined with its low toxicity in nanoparticle form, make it a promising candidate for further clinical trials. The production of anti-microbial and anti-cancer drugs from natural substances, such as chrysin, is a valuable contribution to the field of medicine.

Keywords: chrysin, antimicrobial, anticancer, infectious diseases

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Authors: C. T. Eze, I. N. E. Onwurah

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An important hydrogeologic problem in areas of high faults formations is high environmental health hazard occasioned by microbial and heavy metals contamination of ground waters. Consequently, we examined the microbial load and heavy metals concentration of hospital wastewater discharged into the environment at Park Lane General Hospital Enugu Urban, Nigeria. The microbial counts, characteristics and frequency of occurrences of the isolated microorganisms were determined by cultural, morphological and biochemical characteristics using established procedure while the varying concentrations of the identified heavy metals were determined using the spectrophotometric method. The microbiological analyses showed a mean total aerobic bacteria counts from 13.7 ± 0.65 × 107 to 22.8 ± 1.14 ×1010 CFU/ml, mean total anaerobic bacteria counts from 6.0 ± 1.6 × 103 to 1.7 ± 0.41 ×104 CFU/ml and mean total fungal counts from 0 ± 0 to 2.3 ± 0.16 × 105 CFU/ml. The isolated micro-organisms which included both pathogenic and non-pathogenic organisms were Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Bacillus subtilis, Proteus vulgaris, Klesbsiella pneumonia and bacteriodes sp. The only fungal isolate was Candida albican. The heavy metals identified in the leachate were Arsenic, Cadmium, Lead, Mercury and Chromium and their concentrations ranged from 0.003 ± 0.00082 to 0.14 ± 0.0082 mg/l. These values were above WHO permissible limits while others fall within the limits. Therefore, hospital waste water can pose the environmental health risk when not properly treated before discharge, especially in geologic formations with high fault formations.

Keywords: bacterial isolates, fungal isolates, heavy metals, hospital wastewater, microbial counts

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Authors: Tsige Reda

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Essential oil of Thymus vulgaris was extracted by means of hydro-distillation. This study was done to investigate the chemical composition, antibacterial and antioxidant activities. The chemical composition of the essential oils was determined using gas chromatography coupled to mass spectroscopy (GC-MS). Using disc diffusion assay the antibacterial activity was assessed on one Gram-positive bacteria and one Gram-negative bacteria. The percentage oil yield of the essential oil was found to be 0.97 ± 0.08% (w/w) with yellow color. The physicochemical constants of the oil were also noted. The phytochemical screening of the plant extract revealed the presence of tannins, saponins, phenol, flavonoids, terpenoids, steroids and alkaloids. A total of 18 chemical constituents were identified by Gas Chromatography-Mass Spectroscopy analysis representing 100% of the total essential oil of Thymus vulgaris, with thymol (31.977%), o-cymene (29.992%), and carvacrol (14.541%). Previous studies have revealed that the thymol, o-cymen and carvacrol components of Thymus vulgaris are responsible for their biological activities. Thymus vulgaris have been used traditionally to treat a wide variety of infections. Based on the extensive use and lack of scientific evidence, a study was embarked upon to determine its bioactivity. The essential oil of Thymus vulgaris leaves exhibited higher activity towards the Gram-positive bacteria (Staphylococcus aurous) than the Gram-negative bacteria (Escherichia coli) and also has good antioxidant activity, and can be used medicinal and therapeutic applications. This activity may be due to the high amount of thymol, o-cymen and carvacrol.

Keywords: hydro-distillation, Thymus vulgaris, essential oil composition, phytochemical screening, physicochemical constants, antioxidant activity, antibacterial activity

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Authors: Ali Bagherpour, Masoud Soltanialvar

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Campylobacter species are common bacterial pathogens associated with human gastroenteritis which are generally transmitted through foods of animal origin. This study was carried out to determine the prevalence of Campylobacter species in poultry meat and by products in the city of Dezful in Iran. Since April 2012 to July 2013, a total of 400 samples including meat (n = 100), liver (n = 100), gizzard (n = 100), and poultry heart (n = 100), were randomly collected from Dezful industrial poultry abattoir and were experimented in order to investigate presence of Campylobacter species. According to culture test, 251 samples out of 400 samples under study (69%) were contaminated with Campylobacter species. The highest prevalence of Campylobacter species was observed in poultry's liver (78.3%) and then in gizzard (75.8%), heart (65%) and meat (56.7%). The most common isolated Campylobacter were C. jejuni (90.9%) and the rest were C. coli (9.1%). There was a significant difference (P < 0.05) in the prevalence of Campylobacter species between the meat samples taken in the summer (86.7%). The results of this study indicate the importance of edible offal of poultries as the potential source of Campylobacter infections.

Keywords: Campylobacter jejuni, Campylobacter coli, poultry, meat, products

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Authors: Mohamed Gouda

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Antibacterial wound dressings based on cellulose nanofibers containing different metal nanoparticles (CMC-MNPs) were synthesized using an electrospinning technique. First, the composite of carboxymethyl cellulose containing different metal nanoparticles (CMC/MNPs), such as copper nanoparticles (CuNPs), iron nanoparticles (FeNPs), zinc nanoparticles (ZnNPs), cadmium nanoparticles (CdNPs) and cobalt nanoparticles (CoNPs) were synthesized, and finally, these composites were transferred to the electrospinning process. Synthesized CMC-MNPs were characterized using scanning electron microscopy (SEM) coupled with high-energy dispersive X-ray (EDX) and UV-visible spectroscopy used to confirm nanoparticle formation. The SEM images clearly showed regular flat shapes with semi-porous surfaces. All MNPs were well distributed inside the backbone of the cellulose without aggregation. The average particle diameters were 29-39 nm for ZnNPs, 29-33 nm for CdNPs, 25-33 nm for CoNPs, 23-27 nm for CuNPs and 22-26 nm for FeNPs. Surface morphology, water uptake and release of MNPs from the nanofibers in water and antimicrobial efficacy were studied. SEM images revealed that electrospun CMC-MNPs nanofibers are smooth and uniformly distributed without bead formation with average fiber diameters in the range of 300 to 450 nm. Fiber diameters were not affected by the presence of MNPs. TEM images showed that MNPs are present in/on the electrospun CMC-MNPs nanofibers. The diameter of the electrospun nanofibers containing MNPs was in the range of 300–450 nm. The MNPs were observed to be spherical in shape. The CMC-MNPs nanofibers showed good hydrophilic properties and had excellent antibacterial activity against the Gram-negative bacteria Escherichia coli and the Gram-positive bacteria Staphylococcus aureus.

Keywords: electrospinning technique, metal nanoparticles, cellulosic nanofibers, wound dressing

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Authors: Layal Karam, Rayan Roustom, Mohamad G. Abiad, Tahra El-Obeid, Ioannis N. Savvaidis

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The demand for marinated chicken worldwide, is continuously growing. To date, limited data on addition of active components of Essential Oils (EOs) to marinades for chicken preservation are available. The antimicrobial effect of carvacrol and thymol, added at 0.4 and 0.8% v/w to marinated fresh chicken, stored in air and under vacuum packaging (VP), for 21 days at 4°C, was examined. The samples were monitored for microbiological (total viable count (TVC), lactic acid bacteria (LAB), Brochothrix thermosphacta, Pseudomonas spp., total coliforms, Escherichia coli, yeasts and molds) and sensory attributes (odor characteristics). Our data supports that among the tested microorganisms, Pseudomonas spp., LAB and B. thermosphacta were the most dominant microbiota in the marinated chicken samples. Additionally, the use of active EOs components, especially the higher concentration (0.8% v/w) in combination with VP, retarded the growth of spoilage microbiota and resulted in a significant reduction of about 2.9-3.1 log cfu/g and a microbiological shelf-life extension of marinated chicken by > 6 days, as judged by TVC data. Interestingly, the combination of active components of EOs at the lower concentration (0.4% v/w) and packaging (air or vacuum) resulted in a significant sensorial shelf-life extension of 15 and >21 days, as compared to the controls’ shelf-life of 9 days. The results of our study demonstrated the potential of the active components, carvacrol and thymol, as natural effective antimicrobial hurdles to control the growth of spoilage microorganisms in marinated chicken meat.

Keywords: chicken, essential oils compounds, marination, meat spoilage, preservation

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Authors: Adebowale T. Odeyemi, Oluwafemi A. Ajenifuja

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The leachate samples collected from Erifun dumpsite along Federal Polythenic road, Ado-Ekiti, Ekiti State, were subjected to bacteriological and mineral analyses. The bacteriological estimation and isolation were done using serial dilution and pour plating techniques. Antibiotic susceptibility test was done using agar disc diffusion technique. Atomic Absorption Spectophotometry method was used to analyze the heavy metal contents in the leachate samples. The bacterial and coliform counts ranged from 4.2 × 105 CFU/ml to 2.97 × 106 CFU/ml and 5.0 × 104 CFU/ml to 2.45 x 106 CFU/ml, respectively. The isolated bacteria and percentage of occurrence include Bacillus cereus (22%), Enterobacter aerogenes (18%), Staphylococcus aureus (16%), Proteus vulgaris (14%), Escherichia coli (14%), Bacillus licheniformis (12%) and Klebsiella aerogenes (4%). The mineral value ranged as follow; iron (21.30mg/L - 25.60mg/L), zinc (1.80mg/L - 5.60mg/L), copper (1.00mg/L - 2.60mg/L), chromium (0.50mg/L - 1.30mg/L), candium (0.20mg/L - 1.30mg/L), nickel (0.20mg/L - 0.80mg/L), lead (0.05mg/L-0.30mg/L), cobalt (0.03mg/L - 0.30mg/L) and in all samples manganese was not detected. The entire organisms isolated exhibited a high level of resistance to most of the antibiotics used. There is an urgent need for awareness to be created about the present situation of the leachate in Erifun, on the need for treatment of the nearby stream and other water sources before they can be used for drinking and other domestic use. In conclusion, a good method of waste disposal is required in those communities to prevent leachate formation, percolation, and runoff into water bodies during the raining season.

Keywords: antibiotic susceptibility, dumpsite, bacteriological analysis, heavy metal

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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: Konstans Ruseva, Kristina Ivanova, Katerina Todorova, Margarita Gabrashanska, Tzanko Tzanov, Elena Vassileva

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Zwitterionic polymers (ZP) are well-known with their ultralow biofouling. They are successfully competing with poly(ethylene glycols) (PEG), which are considered as the “golden standard” in this respect. These unique properties are attributed to their strong hydration capacity, defined by the dipole-dipole interactions, arising between the ZP pendant groups as well as to the dipoles interaction with water molecules. Beside, ZP are highly resistant to bacterial adhesion thus ensuring an excellent anti-biofilm formation ability. Moreover, ZP are able to respond upon external stimuli such as temperature, pH, salt concentration changes which in combination with their anti-biofouling effect render this type of polymers as materials with a high potential in biomedical applications. The present work is focused on the development of zwitterionic hydrogels for efficient treatment of highly exudating and hard-to-heal chronic wounds. To this purpose, two types of ZP networks with different crosslinking degree were synthesized - polysulfobetaine (PSB) and polycarboxybetaine (PCB) ones. They were characterized in terms of their physico-mechanical properties, e.g. microhardness, swelling ability, smart behaviour. Furthermore, the potential of ZP networks to resist biofilm formation towards Staphylococcus aureus and Escherichia coli was studied. Their ability to reduce the high levels of myeloperoxidase and metalloproteinase, two enzymes that are part of the chronic wounds enviroenment, was revealed. Moreover, the in vitro cytotoxic assessment of PSB and PCB networks along with their in vivo performance in rats was also studied to reveal their high biocompatibility.

Keywords: absorption properties, biocompatibility, enzymatic inhibition activity, wound healing, zwitterionic polymers

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Authors: Yılmaz Uçar, Fatih Ozogul, Mustafa Durmuş, Yesim Ozogul, Ali Rıza Köşker, Esmeray Kuley Boğa, Deniz Ayas

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The aim of this study was to purified eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), that are essential oils from trout oil, using high-performance liquid chromatography (HPLC) method, bioconverted EPA and DHA into bioconverted EPA (bEPA), bioconverted DHA (bDHA) extracts by P. aeruginosa PR3. Moreover, in vitro antibacterial activity of bEPA and bDHA was investigated using disc diffusion methods and minimum inhibitory concentration (MIC). EPA and DHA concentration of 11.1% and 15.9% in trout oil increased in 58.64% and 40.33% after HPLC optimisation, respectively. In this study, EPA and DHA enriched products were obtained which are to be used as valuable supplements for food and pharmaceutical purposes. The bioconverted EPA and DHA exhibited antibacterial activities against two Gram-positive bacteria (Listeria monocytogenes ATCC 7677 and Staphylococcus aureus ATCC 29213) and six Gram-negative bacteria (Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC700603, Enterococcus faecalis ATCC 29212, Aeromonas hydrophila NCIMB 1135, and Salmonella Paratyphi A NCTC 13). Inhibition zones and MIC value of bEPA and bDHA against bacterial strains ranged from 7 to 12 mm and from 350 to 2350 μg/mL, respectively. Our results suggested that the crude extracts of bioconversion of EPA and DHA by P. aeruginosa PR3 can be considered as promising antimicrobials in improving food safety by controlling foodborne pathogens.

Keywords: High-Performance Liquid Chromatography (HPLC), docosahexaenoic acid, DHA, eicosapentaenoic acid, EPA, minimum inhibitory concentration, MIC, Pseudomonas aeruginosa PR3

Procedia PDF Downloads 468