Search results for: antimicrobial peptides

Authors: Varsha Gupta, Priya Datta, Gursimran Mohi, Jagdish Chander

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Fluoroquinolones form the mainstay of therapy for the treatment of infections due to Salmonella enterica subsp. enterica. There is a complex interplay between several resistance mechanisms for quinolones and various fluoroquinolones discs, giving varying results, making detection and interpretation of fluoroquinolone resistance difficult. For detection of fluoroquinolone resistance in Salmonella ssp., we compared the use of pefloxacin and nalidixic acid discs as surrogate marker. Using MIC for ciprofloxacin as the gold standard, 43.5% of strains showed MIC as ≥1 μg/ml and were thus resistant to fluoroquinoloes. Based on the performance of nalidixic acid and pefloxacin discs as surrogate marker for ciprofloxacin resistance, both the discs could correctly detect all the resistant phenotypes; however, use of nalidixic acid disc showed false resistance in the majority of the sensitive phenotypes. We have also tested newer antimicrobial agents like cefixime, imipenem, tigecycline and azithromycin against Salmonella spp. Moreover, there was a comeback of susceptibility to older antimicrobials like ampicillin, chloramphenicol, and cotrimoxazole. We can also use cefixime, imipenem, tigecycline and azithromycin in the treatment of multidrug resistant S. typhi due to their high susceptibility.

Keywords: salmonella, pefloxacin, surrogate marker, chloramphenicol

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Authors: M.Ayazi, N. Golshan Ebrahimi

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The development of antibacterial surface has devoted extensive researches and important field due to the growing antimicrobial resistance strains. The superhydrophobic surface has raised attention because of reducing bacteria adhesion in the absence of antibiotic agents. Evaluating the current development antibacterial surface has to be investigating to consider the potential of applying superhydrophobic surface to reduce bacterial adhesion or role of patterned surfaces on it. In this study, we present different samples with bio-inspired hierarchical and microstructures to consider their ability in reducing bacterial adhesion. The structures have inspired from rice-like pattern and lotus-leaf that developed on the polydimethylsiloxane (PDMS) and polypropylene (PP). The results of the attachment behaviors have considered on two bacteria strains of gram-negative Escherichia coli (E. coli) bacteria and gram-positive Staphylococcus aureus (S. aureus). The reduction of bacteria adhesion on these roughness surfaces demonstrated the effectiveness of rinsing ability on removing bacterial cells on structured plastic surfaces. Results have also offered the important role of bacterial species, material chemistry and hierarchical structure to prevent bacterial adhesion.

Keywords: hierarchical structure, self-cleaning, lotus-effect, bactericidal

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Authors: Faranak Beigmohammadi, Seyed Hadi Peighambardoust, Seyed Jamaledin Peighambardoust

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This study undertakes to investigate the ability of different kinds of nanocomposite films made of cloisite-30B with different percentages of silver and copper oxide nanoparticles incorporated into a low-density polyethylene (LDPE) polymeric matrix by a melt mixing method in order to inhibit the growth of microorganism in soy burger. The number of surviving cell of the total count was decreased by 3.61 log and mold and yeast diminished by 2.01 log after 8 weeks storage at 18 ± 0.5°C below zero, whilst pure LDPE did not has any antimicrobial effect. A composition of 1.3 % cloisite 30B-Ag and 2.7 % cloisite 30B-CuO for total count and 0 % cloisite 30B-Ag and 4 % cloisite 30B-CuO for yeast & mold gave optimum points in combined design test in Design Expert 7.1.5. Suitable microbial models were suggested for retarding above microorganisms growth in soy burger. To validation of optimum point, the difference between the optimum point of nanocomposite film and its repeat was not significant (p<0.05) by one-way ANOVA analysis using SPSS 17.0 software, while the difference was significant for pure film. Migration of metallic nanoparticles into a food stimulant was within the accepted safe level.

Keywords: modeling, nanocomposite film, packaging, soy burger

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Authors: Abdurrahim A. Elouzi, Abdurrauf M. Gusbi, Ali M. Elgerbi

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Antibiotic resistance has become a global public-health problem, thus it is imperative that new antibiotics continue to be developed. Major problems are being experienced in human medicine from antibiotic resistant bacteria. Moreover, no new chemical class of antibiotics has been introduced into medicine in the past two decades. The aim of the current study presents experimental results that evaluate the capability of bio surfactant rhamnolipid on enhancing the efficacy of hydrophobic antibiotics. Serial dilutions of azithromycin and clarithromycin were prepared. A bacterial suspension (approximately 5 X 105 CFU) from an overnight culture in MSM was inoculated into 20 ml sterile test tube each containing a serial 10-fold dilution of the test antibiotic(s) in broth with or without 200 mgL-1 rhamnolipid. The tubes were incubated for 24 h with vigorous shaking at 37°C. Antimicrobial activity in multiple antibiotic-resistant gram-negative bacteria pathogens and gram-positive bacteria were assessed using optical density technique. The results clearly demonstrated that the presence of rhamnolipid significantly improved the efficiency of both antibiotics. We hypothesized that the addition of rhamnolipid at low concentration, causes release of LPS which results in an increase in cell surface hydrophobicity. This allows increased association of cells with hydrophobic antibiotics resulting in increased cytotoxicity rates.

Keywords: hydrophobic antibiotics, biosurfactant, rhamnolipid, azithromycin, clarithromycin

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Authors: Ankita Baidya, Vanishri Ganakumar, Ranveer S. Jadon, Piyush Ranjan, Rita Sood

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Septic embolism can have varied presentations and clinical considerations. Infected central venous catheters are commonly associated with septic emboli but peripheral vascular catheters are rarely implicated. We describe a rare case of septic pulmonary emboli related to infected peripheral venous cannulation caused by an unusual etiological agent. A young male presented with complaints of fever, productive cough, sudden onset shortness of breath and cellulitis in both the upper limbs. He was recently hospitalised for dengue fever and administered intravenous fluids through peripheral venous line. The patient was febrile, tachypneic and in respiratory distress, there were multiple pus filled bullae in left hand alongwith swelling and erythema involving right forearm that started at the site of cannulation. Chest examination showed active accessory muscles of respiration, stony dull percussion at the base of right lung and decreased breath sounds at right infrascapular, infraaxillary and mammary area. Other system examination was within normal limits. Chest X-ray revealed bilateral multiple patchy heterogenous peripheral opacities and infiltrates with right-sided pleural effusion. Contrast-enhanced computed tomography (CECT) chest showed feeding vessel sign confirming the diagnosis as septic emboli. Venous Doppler and 2D-echocardiogarm were normal. Laboratory findings showed marked leucocytosis (22000/mm3). Pus aspirate, blood sample, and sputum sample were sent for microbiological testing. The patient was started empirically on ceftriaxone, vancomycin, and clindamycin. The Pus culture and sputum culture showed Klebsiella pneumoniae sensitive to cefoperazone-sulbactum, piperacillin-tazobactum, meropenem and amikacin. The antibiotics were modified accordingly to antimicrobial sensitivity profile to Cefoperazone-sulbactum. Bronchoalveolar lavage (BAL) was done and sent for microbiological investigations. BAL culture showed Klebsiella pneumoniae with same antimicrobial resistance profile. On day 6 of starting cefoperazone-sulbactum, he became afebrile. The skin lesions improved significantly. He was administered 2 weeks of cefoperazone–sulbactum and discharged on oral faropenem for 4 weeks. At the time of discharge, TLC was 11200/mm3 with marked radiological resolution of infection and healed skin lesions. He was kept in regular follow up. Chest X-ray and skin lesions showed complete resolution after 8 weeks. Till date, only couple of case reports of septic emboli through peripheral intravenous line have been reported in English literature. This case highlights that a simple procedure of peripheral intravenous cannulation can lead to catastrophic complication of septic pulmonary emboli and widespread cellulitis if not done with proper care and precautions. Also, the usual pathogens in such clinical settings are gram positive bacteria, but with the history of recent hospitalization, empirical therapy should also cover drug resistant gram negative microorganisms. It also emphasise the importance of appropriate healthcare practices to be taken care during all procedures.

Keywords: antibiotics, cannula, Klebsiella pneumoniae, septic emboli

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Authors: Kinza Khan, Dad Muhmmad, Asif Saleem, Nadia Mukhtar, Tahir Yaqub

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Ethomedicines are more commonly used in underdeveloped and developing countries. These medicines are sometimes more potent in controlling microbial infections than conventional medicines. Medicinal plants have been common practice to cure many diseases for centuries. Murraya koenigii is one of these plants and is commonly used in South Asian countries as a flavoring agent in food. To evaluate its anti-microbial activity, six different bacterial strains (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhi, Bacillus cereus and Klebsiella pneumonia were used. N-hexane extract of Murraya koenigii leaves shows maximum activity against Bacillus cereus. Acetone extract of Murraya koenigii shoots showed more efficient activity against Pseudomonas aeruginosa Dichloromethane extracts showed maximum activity against Bacillus cereus. Ethanol extract exhibited maximum activity against Pseudomonas aeruginosa and Klebsiella pneumoniae. The methanol extract of Murraya koenigii shoots displayed maximum antibacterial activity against Bacillus cereus. Antifungal activity Ethanol extract was more effective against Candida albicans.

Keywords: ethnomedicines, bacteria, fungi, murraya koenigii, antimicrobial activity

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Authors: Ali Amiri, Arash Esfandiari, Elham Zarenezhad

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We report here an efficient and rapid method for the preparation of 3,4-dihydro-1H-quinoxalin-2-ones and 1H‑quinolin-2-ones that involves grinding of o-, m-, or p‑phenylenediamine and three dialkyl acetylenedicarboxylates using a pestle and mortar. This solvent-free approach requires only a few minutes of reaction time. This type of reaction is expected to be the most economical method since neither catalyst nor solvent is used. Finally, all synthesised compounds were screened for antimicrobial activity against two Gram-positive bacteria (Pseudomonas aeruginosa PTCC 1077, Escherichia coli PTCC1330) and two Gram-negative bacteria (Staphylococcus aureus PTCC 1133, Bacillus cereus PTCC 1015) and their activity. Compared with gentamycin and ampicillin as reference drugs for Gram-negative and Gram-positive bacteria, respectively. The minimum inhibitory concentration (MIC) of the synthesised compounds and reference drugs were determined by the microdilution method. Good antibacterial activity was observed for 3,4-dihydro-1H-quinoxalin-2-ones against all species of Gram-positive and Gram-negative bacteria, and1H‑quinolin-2-ones showed good antibacterial activity against two Gram-positive bacteria.

Keywords: quinolin, quinoxalin, anti-bacterial activity, minimum inhibitory concentration (MIC)

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Authors: S. Fröhlich, M. Herold, M. Allmer

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Early stage quantitative analysis of host cell protein (HCP) variations is challenging yet necessary for comprehensive bioprocess development. High resolution mass spectrometry (HRMS) provides a high-end technology for accurate identification alongside with quantitative information. Hereby we describe a flexible HRMS assay platform to quantify HCPs relevant in microbial expression systems such as E. Coli in both up and downstream development by means of MVDA tools. Cell pellets were lysed and proteins extracted, purified samples not further treated before applying the SMART tryptic digest kit. Peptides separation was optimized using an RP-UHPLC separation platform. HRMS-MSMS analysis was conducted on an Orbitrap Velos Elite applying CID. Quantification was performed label-free taking into account ionization properties and physicochemical peptide similarities. Results were analyzed using SIEVE 2.0 (Thermo Fisher Scientific) and SIMCA (Umetrics AG). The developed HRMS platform was applied to an E. Coli expression set with varying productivity and the corresponding downstream process. Selected HCPs were successfully quantified within the fmol range. Analysing HCP networks based on pattern analysis facilitated low level quantification and enhanced validity. This approach is of high relevance for high-throughput screening experiments during upstream development, e.g. for titer determination, dynamic HCP network analysis or product characterization. Considering the downstream purification process, physicochemical clustering of identified HCPs is of relevance to adjust buffer conditions accordingly. However, the technology provides an innovative approach for label-free MS based quantification relying on statistical pattern analysis and comparison. Absolute quantification based on physicochemical properties and peptide similarity score provides a technological approach without the need of sophisticated sample preparation strategies and is therefore proven to be straightforward, sensitive and highly reproducible in terms of product characterization.

Keywords: process analytical technology, mass spectrometry, process characterization, MVDA, pattern recognition

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Authors: Mehani Mouna, Boukhari Nadjet, Ladjal Segni

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The aim of our study is to determine the antimicrobial effect of essential oils of two plants Cotula cinerea and Chamomilla recutita on some pathogenic bacteria. It is a medicinal plant used in traditional therapy. Essential oils have many therapeutic properties. In herbal medicine, they are used for their antiseptic properties against infectious diseases of fungal origin, against dermatophytes, those of bacterial origin. Essential oils have many therapeutic properties. In herbal medicine, they are used for their antiseptic properties against infectious diseases of fungal origin, against dermatophytes, those of bacterial origin. Humans use plants for thousands of years to treat various ailments, in many developing countries; much of the population relies on traditional doctors and their collections of medicinal plants to cure them. The test adopted is based on the diffusion method on solid medium (Antibiogram), this method allows to determine the susceptibility or resistance of an organism according to the sample studied. Our study reveals that the essential oil of the plants Cotula cinerea and Chamomilla recutita have a different effect on the resistance of germs.

Keywords: antibiogram, Chamomilla recutita, Cotula cinerea, essential oil, microorganism

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Authors: Mateen A Khan, Taj Mohammad, Md. Imtaiyaz Hassan

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Alzheimer’s disease is characterized by the accumulation of the processing products of the amyloid beta peptide cleaved by amyloid precursor protein (APP). Iron increases the synthesis of amyloid beta peptides, which is why iron is present in Alzheimer's disease patients' amyloid plaques. Iron misregulation in the brain is linked to the overexpression of APP protein, which is directly related to amyloid-β aggregation in Alzheimer’s disease. The APP 5'-UTR region encodes a functional iron-responsive element (IRE) stem-loop that represents a potential target for modulating amyloid production. Targeted regulation of APP gene expression through the modulation of 5’-UTR sequence function represents a novel approach for the potential treatment of AD because altering APP translation can be used to improve both the protective brain iron balance and provide anti-amyloid efficacy. The molecular docking analysis of APP IRE RNA with eukaryotic translation initiation factors yields several models exhibiting substantial binding affinity. The finding revealed that the interaction involved a set of functionally active residues within the binding sites of eIF4F. Notably, APP IRE RNA and eIF4F interaction were stabilized by multiple hydrogen bonds with residues of APP IRE RNA and eIF4F. It was evident that APP IRE RNA exhibited a structural complementarity that tightly fit within binding pockets of eIF4F. The simulation studies further revealed the stability of the complexes formed between RNA and eIF4F, which is crucial for assessing the strength of these interactions and subsequent roles in the pathophysiology of Alzheimer’s disease. In addition, MD simulations would capture conformational changes in the IRE RNA and protein molecules during their interactions, illustrating the mechanism of interaction, conformational change, and unbinding events and how it may affect aggregation propensity and subsequent therapeutic implications. Our binding studies correlated well with the translation efficiency of APP mRNA. Overall, the outcome of this study suggests that the genomic modification and/or inhibiting the expression of amyloid protein by targeting APP IRE RNA can be a viable strategy to identify potential therapeutic targets for AD and subsequently be exploited for developing novel therapeutic approaches.

Keywords: Alzheimer's disease, Protein-RNA interaction analysis, molecular docking simulations, conformational dynamics, binding stability, binding kinetics, protein synthesis.

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Authors: L. Szymanski, Z. Kolacinski, Z. Kamiński, G. Raniszewski, J. Fraczyk, L. Pietrzak

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In the article the development and demonstration of method and the model device for hyperthermic selective destruction of cancer cells are presented. This method was based on the synthesis and functionalization of carbon nanotubes serving as ferromagnetic material nano containers. Methodology of the production carbon - ferromagnetic nanocontainers includes: the synthesis of carbon nanotubes, chemical and physical characterization, increasing the content of ferromagnetic material and biochemical functionalization involving the attachment of the key addresses. Biochemical functionalization of ferromagnetic nanocontainers is necessary in order to increase the binding selectively with receptors presented on the surface of tumour cells. Multi-step modification procedure was finally used to attach folic acid on the surface of ferromagnetic nanocontainers. Folic acid is ligand of folate receptors which is overexpresion in tumor cells. The presence of ligand should ensure the specificity of the interaction between ferromagnetic nanocontainers and tumor cells. The chemical functionalization contains several step: oxidation reaction, transformation of carboxyl groups into more reactive ester or amide groups, incorporation of spacer molecule (linker), attaching folic acid. Activation of carboxylic groups was prepared with triazine coupling reagent (preparation of superactive ester attached on the nanocontainers). The spacer molecules were designed and synthesized. In order to ensure biocompatibillity of linkers they were built from amino acids or peptides. Spacer molecules were synthesized using the SPPS method. Synthesis was performed on 2-Chlorotrityl resin. The linker important feature is its length. Due to that fact synthesis of peptide linkers containing from 2 to 4 -Ala- residues was carried out. Independent synthesis of the conjugate of foilic acid with 6-aminocaproic acid was made. Final step of synthesis was connecting conjugat with spacer molecules and attaching it on the ferromagnetic nanocontainer surface. This article contains also information about special CVD and microvave plasma system to produce nanotubes and ferromagnetic nanocontainers. The first tests in the device for hyperthermal RF generator will be presented. The frequency of RF generator was in the ranges from 10 to 14Mhz and from 265 to 621kHz.

Keywords: synthesis of carbon nanotubes, hyperthermia, ligands, carbon nanotubes

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Authors: Milica Z. Karadzic, Lidija R. Jevric, Sanja Podunavac-Kuzmanovic, Strahinja Z. Kovacevic, Anamarija I. Mandic, Katarina Penov-Gasi, Andrea R. Nikolic, Aleksandar M. Okljesa

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In this study, a set of 29 newly synthesized steroid derivatives were investigated using reversed-phase high-performance liquid chromatography (RP-HPLC) as a first step in preselection of drug candidates. This analysis presents an experimental determination of chromatographic lipophilicity, and it was conducted to obtain physicochemical characterization of these molecules. As the most widely used bonded phases in RP-HPLC, octadecyl (C18) and octyl (C8) were used. Binary mixtures of water and acetonitrile or methanol were used as mobile phases. Obtained results were expressed as retention factor values logk and they were correlated with logP values. The results showed that both columns provide good estimations of the chromatographic lipophilicity of the molecules included in this study. This analysis was conducted in order to characterize newly synthesized steroid derivatives for further investigation regarding their antiproliferative and antimicrobial activity. This article is based upon work from COST Action (CM1306), supported by COST (European Cooperation in Science and Technology).

Keywords: antiproliferative activity, chromatographic lipophilicity, liquid chromatography, steroids

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Authors: Nuray Yılmaz Baran

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The conjugated Schiff base polymers which are also called as polyazomethines are promising materials for various applications due to their good thermal resistance semiconductive, liquid crystal, fiber forming, nonlinear optical outstanding photo- and electroluminescence and antimicrobial properties. In recent years, polyazomethines have attracted intense attention of researchers especially due to optoelectronic properties which have made its usage possible in organic light emitting diodes (OLEDs), solar cells (SCs), organic field effect transistors (OFETs), and photorefractive holographic materials (PRHMs). In this study, N-(pyridin-2-ylmethylidene)pyridin-2-amine Schiff base was synthesized from condensation reaction of 2-aminopyridine with 2-pyridine carbaldehyde. Polymerization of Schiff base was achieved by polycondensation reaction using NaOCl oxidant in methanol medium at various time and temperatures. The synthesized Schiff base monomer and polymer (Poly(N-(pyridin-2-ylmethylidene)pyridin-2-amine)) was characterized by UV-vis, FT-IR, 1H-NMR, XRD techniques. Molecular weight distribution and the surface morphology of the polymer was determined by GPC and SEM-EDAX techniques. Thermal behaviour of the monomer and polymer was investigated by TG/DTG, DTA and DSC techniques.

Keywords: polyazomethines, polycondensation reaction, Schiff base polymers, thermal stability

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Authors: Borokini Funmilayo Boede

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B. alba L., commonly called ‘Amunututu’ and Solanecio biafrae called ‘Worowo’ among the Yoruba tribe in the southwest part of Nigeria are reported to be of great ethnomedicinal importance but are among many underutilized green leafy vegetables in the country. Many studies have established the nutritional values of these vegetables, utilization are very poor and indepth information on their chemical profiles is scarce. The aqueous, methanolic and ethanolic extracts of these vegetables were subjected to phytochemical screening and phenolic profiles of the alcoholic extracts were characterized by using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD). Total phenol and flavonoid contents were determined, antioxidant activities were evaluated using five in vitro assays to assess DPPH, nitric oxide and hydroxyl radical-scavenging abilities, as well as reducing power with ferric reducing antioxidant assay and phosphomolybdate method. The antibacterial activities of the extracts against Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella typhi were evaluated by using agar well diffusion method and the antifungal activity evaluated against food-associated filamentous fungi by using poisoned food technique with the aim of assessing their nutraceutical potentials to encourage their production and utilization. The results revealed the presence of saponnin, steroids, tannin, terpenoid and flavonoid as well as phenolic compounds: gallic acid, chlorogenic acid, caffeic acid, coumarin, rutin, quercitrin, quercetin and kaemferol. The vegetables showed varying concentration dependent reducing and radical scavenging abilities from weak to strong compared with gallic acid, rutin, trolox and ascorbic acid used as positive controls; the aqueous extracts which gave higher concentrations of total phenol displayed higher ability to reduce Fe (lll) to Fe (ll) and stronger inhibiting power against hydroxyl radical than the alcoholic extracts and in most cases exhibited more potency than the ascorbic acids used as positive controls, at the same concentrations, whereas, methanol and / or ethanol extracts were found to be more effective in scavenging 2, 2-diphenyl-1-picryl hydrazyl radical and showed higher ability to reduce Mo (VI) to Mo (V) in total antioxidant assay than the aqueous extracts. However, the inhibition abilities of all the extracts against nitric oxide were comparable with the ascorbic acid control at the same concentrations. There were strong positive correlations with total phenol (mg GAE/g) and total flavonoid (mg RE/g) contents in the range TFC (r=0.857- 0999 and r= 0.904-1.000) and TPC (r= 0.844- 0.992 and r= 0.900 -0.999) for Basella alba and Senecio biafrae respectively. Inhibition concentration at 50 % (IC50) for each extract to scavenge DPPH, OH and NO radicals ranged from 32.73 to 1.52 compared with control (0.846 - -6.42) mg/ml. At 0.05g/ml, the vegetables were found to exhibit mild antibacterial activities against Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella typhi compared with streptomycin sulphate used as control but appreciable antifungi activities against (Trichoderma rubrum and Aspergillus fumigates) compared with bonlate antibiotic positive control. The vegetables possess appreciable antioxidant and antimicrobial properties for promoting good health, their cultivation and utilization should be encouraged especially in the face of increasing health and economic challenges and food insecurity in many parts of the world.

Keywords: antimicrobial, antioxidants, extracts, phytochemicals

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Authors: Vandana Deopanée Tomar, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra, Sandhya Mehrotra

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The cyanobacterial CO₂ concentrating mechanism (CCM) operates to raise the levels of CO₂ in the vicinity of the main carboxylation enzyme Rubisco which is encapsulated in protein micro compartments called carboxysomes. Thus, due to the presence of CCM, cyanobacterial cells are able to work with high photosynthetic efficiency even at low Ci conditions and can accumulate 1000 folds high internal concentrations of Ci than external environment. Engineering of some useful CCM components into higher plants is one of the plausible approaches to improve their photosynthetic performance. The first step and the simplest approach for attaining this objective would be the transfer of cyanobacterial bicarbonate transporter such as SbtA to inner chloroplast envelope of C₃ plants. For this, SbtA transporter gene from Synechococcus elongatus PCC 7942 was fused to a transit peptide element to generate chimeric constructs in order to direct it to chloroplast inner envelope. Two transit peptides namely, TnaXTP (transit peptide from AT3G56160) and TMDTP (transit peptide from AT2G02590) were shortlisted from Arabidopsis thaliana genome and cloned in plant expression vector pCAMBIA1302 having mgfp5 as a reporter gene. Plant transformation was done by agro infiltration and Agrobacterium mediated co-culture. DNA, RNA, and protein were isolated from the leaves four days post infiltration, and the presence of transgene was confirmed by gene specific PCR (Polymerase Chain Reaction) analysis and by RT-PCR (Reverse Transcription Polymerase Chain Reaction). The expression was confirmed at the protein level by western blotting using anti-GFP primary antibody and horseradish peroxidase (HRP) conjugated secondary antibody. The localization of the protein was detected by confocal microscopy of isolated protoplasts. We observed chloroplastic expression for both the fusion constructs which suggest that the transit peptide sequences are capable of taking the cargo protein to the chloroplasts. These constructs are now being used to generate stable transgenic plants by Agrobacterium mediated transformation. The stability of transgene expression will be analyzed from T₀ to T₂ generation.

Keywords: agro infiltration, bicarbonate transporter, carbon concentrating mechanisms, cyanobacteria, SbtA

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Authors: Lee Yujin, Han Jiye, Oh Jin-Woo

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The adverse effects of endocrine disrupting chemicals (EDCs) has attracted considerable public interests. The benzene-like EDCs structure mimics the mechanisms of hormones naturally occurring in vivo, and alters physiological function of the endocrine system. Although, some of the most representative EDCs such as polychlorinated biphenyls (PCBs) and phthalates compounds already have been prohibited to produce and use in many countries, however, PCBs and phthalates in plastic products as flame retardant and plasticizer are still circulated nowadays. EDCs can be released from products while using and discarding, and it causes serious environmental and health issues. Here, we developed virus-based structurally coloured nanostructure that can detect minute EDCs concentration sensitively and selectively. These structurally coloured nanostructure exhibits characteristic angel-independent colors due to the regular virus bundle structure formation through simple pulling technique. The designed number of different colour bands can be formed through controlling concentration of virus solution and pulling speed. The virus, M-13 bacteriophage, was genetically engineered to react with specific ECDs, typically PCBs and phthalates. M-13 bacteriophage surface (pVIII major coat protein) was decorated with benzene derivative binding peptides (WHW) through phage library method. In the initial assessment, virus-based color sensor was exposed to several organic chemicals including benzene, toluene, phenol, chlorobenzene, and phthalic anhydride. Along with the selectivity evaluation of virus-based colour sensor, it also been tested for sensitivity. 10 to 300 ppm of phthalic anhydride and chlorobenzene were detected by colour sensor, and showed the significant sensitivity with about 90 of dissociation constant. Noteworthy, all measurements were analyzed through principal component analysis (PCA) and linear discrimination analysis (LDA), and exhibited clear discrimination ability upon exposure to 2 categories of EDCs (PCBs and phthalates). Because of its easy fabrication, high sensitivity, and the superior selectivity, M-13 bacteriophage-based color sensor could be a simple and reliable portable sensing system for environmental monitoring, healthcare, social security, and so on.

Keywords: M-13 bacteriophage, colour sensor, genetic engineering, EDCs

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Authors: D. Malouch, M. Berchel, C. Dreanno, S. Stachowski-Haberkorn, P-A. Jaffres

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Biofilm is a predominant lifestyle chosen by bacteria. Whether it is developed on an immerged surface or a mobile biofilm known as flocs, the bacteria within this form of life show properties different from its planktonic ones. Within the biofilm, the self-formed matrix of Extracellular Polymeric Substances (EPS) offers hydration, resources capture, enhanced resistance to antimicrobial agents, and allows cell-communication. Biofouling is a complex natural phenomenon that involves biological, physical and chemical properties related to the environment, the submerged surface and the living organisms involved. Bio-colonization of artificial structures can cause various economic and environmental impacts. The increase in costs associated with the over-consumption of fuel from biocolonized vessels has been widely studied. Measurement drifts from submerged sensors, as well as obstructions in heat exchangers, and deterioration of offshore structures are major difficulties that industries are dealing with. Therefore, surfaces that inhibit biocolonization are required in different areas (water treatment, marine paints, etc.) and many efforts have been devoted to produce efficient and eco-compatible antifouling agents. The different steps of surface fouling are widely described in literature. Studying the biofilm and its stages provides a better understanding of how to elaborate more efficient antifouling strategies. Several approaches are currently applied, such as the use of biocide anti-fouling paint6 (mainly with copper derivatives) and super-hydrophobic coatings. While these two processes are proving to be the most effective, they are not entirely satisfactory, especially in a context of a changing legislation. Nowadays, the challenge is to prevent biofouling with non-biocide compounds, offering a cost effective solution, but with no toxic effects on marine organisms. Since the micro-fouling phase plays an important role in the regulation of the following steps of biofilm formation7, it is desired to reduce or delate biofouling of a given surface by inhibiting the micro fouling at its early stages. In our recent works, we reported that some amphiphilic compounds exhibited bacteriostatic or bactericidal properties at a concentration that did not affect eukaryotic cells. These remarkable properties invited us to assess this type of bio-inspired phospholipids9 to prevent the colonization of surfaces by marine bacteria. Of note, other studies reported that amphiphilic compounds interacted with bacteria leading to a reduction of their development. An amphiphilic compound is a molecule consisting of a hydrophobic domain and a polar head (ionic or non-ionic). These compounds appear to have interesting antifouling properties: some ionic compounds have shown antimicrobial activity, and zwitterions can reduce nonspecific adsorption of proteins. Herein, we investigate the potential of amphiphilic compounds as inhibitors of bacterial growth and marine biofilm formation. The aim of this study is to compare the efficacy of four synthetic phospholipids that features a cationic charge (BSV36, KLN47) or a zwitterionic polar-head group (SL386, MB2871) to prevent microfouling with marine bacteria. We also study the toxicity of these compounds in order to identify the most promising compound that must feature high anti-adhesive properties and a low cytotoxicity on two links representative of coastal marine food webs: phytoplankton and oyster larvae.

Keywords: amphiphilic phospholipids, bacterial biofilm, marine microfouling, non-toxic antifouling

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Authors: Daria Olkiewicz, Maciej Walczak

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In this paper, the biocidal effects of trans-cinnamaldehyde (a main component of cinnamon oil) and geraniol (a constituent of Pelargonium graveolens essential oil) are presented. The activity of the combination of trans-cinnamaldehyde and geraniol was tested against 3 bacterial strains: Staphylococcus aureus ATCC6538 (Gramm+), Escherichia coli ATCC8739 (Gramm-, Lac+) and Pseudomonas aeruginosa KKP 991(Gramm-, Lac-). The biocidal activity of trans-cinnamaldehyde-geraniol mixture against bacteria mentioned above was evaluated by disk-diffusion method. The model strains were exposed on 1, 2.5, 5 and 10 mg of trans-cinnamaldehyde-geraniol mixture per disk, and all strains were susceptible to this combination of plant compounds. For all microorganisms, also Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) were estimated. For Staphylococcus aureus MIC was 0.0625 mg/ml of the trans-cinnamaldehyde and geraniol mixture, and MBC was 1.25 mg/ml; For Escherichia coli MIC=0.5 mg/ml, MBC=1 mg/ml, and finally Pseudomonas aeruginosa was inhibited in 0.5 mg/ml, and minimal biocidal concentration of tested mixture for it was 1.25 mg/ml. There are also reports about the synergistic working of trans-cinnamaldehyde and geraniol against microorganisms and the antimicrobial activity of polymers enriched with trans-cinnamaldehyde or geraniol, therefore the successful development and introduction to the today life of biocidal polymer enriched with trans-cinnamaldehyde and geraniol are possible.

Keywords: antibacterial activity, biocidal polymers, geraniol, trans-cinnamaldehyde

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Authors: Kamel A. Abd-Elsalam, Alexei R. Khokhlov

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Studies on bioefficacy (in vitro and in vivo) and mode of action of the nanocides against the most important plant diseases in Egypt and Russia might assist in the goal of sustainable agriculture. To our knowledge, few researchers have evaluated the combined antimicrobial effect of inorganic nanoparticles (NPs) with bioorganic pesticides for controlling plant pathogens in the greenhouse and open field, decontrol investigated synergistic effect. In the current project, we will develop eco-friendly alternative management strategies including the use of heavy nanometal-tolerant Trichoderma strains and the main effective material in conventional fungicides (curpic, sulfur, phosphorus and zinc) for controlling plant diseases. Studies on bioefficacy and the mechanism of the nanocides against the most important plant diseases in Egypt were evaluated. There is a growing need to establish mechanisms of action for nano bio and/or fungicides to assist the design of new compounds or combinations of compounds, in order to understand resistance mechanisms and to provide a focus for toxicological attention. Nanofungicides represent an emerging technological development that could offer a range of benefits including increased efficacy, durability, and a reduction in the amounts of active ingredients that need to be used.

Keywords: biohybrids, biocides, bioagent, plant pathogenic fungi

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Authors: Kiran Fatima, Kashif Ali

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Staphylococcus aureus is an opportunistic human as well as animal pathogen that causes a variety of diseases. A total of 70 staphylococci isolates were obtained from soil, water, yogurt, and clinical samples. The likely staphylococci clinical isolates were identified phenotypically by different biochemical tests. Molecular identification was done by PCR using species-specific 16S rRNA primer pairs, and finally, 50 isolates were found to be positive as Staphylococcus aureus, sciuri, xylous and cohnii. Screened isolates were further analyzed by several microbiological diagnostics tests, including gram staining, coagulase, capsule, hemolysis, fermentation of glucose, lactose, maltose, and sucrose tests enzymatic reactions. It was found that 78%, 81%, and 51% of isolates were positive for gelatin hydrolysis, protease, and lipase activities, respectively. Antibiogram analysis of isolated Staphylococcus aureus strains with respect to different antimicrobial agents revealed resistance patterns ranging from 57 to 96%. Our study also shows 70% of strains to be MRSA, 54.3% as VRSA, and 54.3% as both MRSA and VRSA. All the identified isolates were subjected to detection of mecA, nuc, and hlb genes, and 70%, 84%, and 40% were found to harbour mecA, nuc, and hlb genes, respectively. The current investigation is highly important and informative for the high-level multidrug-resistant Staphylococcus aureus infections inclusive also of methicillin and vancomycin.

Keywords: MRSA, VRSA, mecA, MSSA

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Authors: Amir Amiri, Naghmeh Morakabati

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During the recent years the six-fold growth of cancer in Iran has led the production of healthy products to become a challenge in the food industry. Due to the young population in the country, the consumption of fast foods is growing. The chemical cancer-causing preservatives are used to produce these products more than the standard; so using an appropriate alternative seems to be important. On the one hand, the plant essential oils show the high antimicrobial potential against pathogenic and spoilage microorganisms and on the other hand they are highly volatile and decomposed under the processing conditions. The study aims to produce the loaded chitosan nanoparticles with different concentrations of savory essential oil to improve the anti-microbial property and increase the resistance of essential oil to oxygen and heat. The encapsulation efficiency was obtained in the range of 32.07% to 39.93% and the particle size distribution of the samples was observed in the range of 159 to 210 nm. The range of Zeta potential was obtained between -11.9 to -23.1 mV. The essential oil loaded in chitosan showed stronger antifungal activity against Rhizopus stolonifer. The results showed that the antioxidant property is directly related to the concentration of loaded essential oil so that the antioxidant property increases by increasing the concentration of essential oil. In general, it seems that the savory essential oil loaded in chitosan particles can be used as a food processor.

Keywords: chitosan, encapsulation, essential oil, nanogel

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Authors: Hulya Tuba Kiyan, Nilgun Ozturk

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Gentiana, a member of Gentianaceae, is represented by approximately 400 species in the world and 12 species in Turkey. Flavonoids, iridoids, triterpenoids and also xanthones are the major compounds of this genus, have been previously reported to have antiinflammatory, antimicrobial, antioxidant, hepatoprotective, hypotensive, hypoglycaemic, DNA repair and immunomodulatory properties. The methanolic extract of the aerial parts of Gentiana brachyphylla Vill. from Turkey was evaluated for its biological activities and its total phenolic content in the present study. According to the antioxidant activity results, G. brachyphylla methanolic extract showed very strong anti-DNA damage antioxidant activity with an inhibition of 81.82%. It showed weak ferric-reducing power with a EC50 value of 0.65 when compared to BHT (EC50 = 0.2). Also, at 0.5 mg/ml concentration, the methanolic extract inhibited ABTS radical cation activity with an inhibition of 20.13% when compared to Trolox (79.01%). Chelating ability of G. brachyphylla was 44.71% whereas EDTA showed 78.87% chelating activity at 0.2 mg/ml. Also G. brachyphylla showed weak 27.21% AChE, 20.23% BChE, strong 67.86% MAO-A and moderate 50.06% MAO-B, weak 19.14% COX-1, 29.11% COX-2 inhibitory activities at 0.25 mg/ml. The total phenolic content of G. brachyphylla was 156.23 ± 2.73 mg gallic acid equivalent/100 g extract.

Keywords: antioxidant activity, cholinesterase inhibitory activity, Gentiana brachyphylla Vill., total phenolic content

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Authors: Simona Dostalova, Dita Munzova, Ana Maria Jimenez Jimenez, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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The boom of nanomedicine in recent years has led to the development of numerous new nanomaterials that can be used as nanocarriers in the drug delivery. These nanocarriers can either be synthetic or natural-based. The disadvantage of many synthetic nanocarriers is their toxicity in patient’s body. Protein cages that can naturally be found in human body do not exhibit such disadvantage. However, the release of cargo from some protein cages in target cells can be problematic. As a special type of protein cages can serve the capsid of many viruses, including phage. Phages infect bacterial cells; therefore they are not harmful to human cells. The targeting of phage particles to cancer cells can be solved by producing of empty phage capsids during which the targeting moieties (e.g. peptides) can be cloned into genes of phage capsid to decorate its surface. Moreover, the produced capsids do not contain viral nucleic acid and are therefore not infectious to beneficial bacteria in the patient’s body. The protein cage composed of viral capsid is larger than other frequently used apoferritin cage but its size is still small enough to benefit from passive targeting by Enhanced Permeability and Retention effect. In this work, bacteriophage λ was used, both whole and its empty capsid for delivery of different cytotoxic drugs (cisplatin, carboplatin, oxaliplatin, etoposide and doxorubicin). Large quantities of phage λ were obtained from phage λ-producing strain of E. coli cultivated in medium with 0.2 % maltose. After killing of E. coli with chloroform and its removal by centrifugation, the phage was concentrated by ultracentrifugation at 130 000 g and 4 °C for 3 h. The encapsulation of the drugs was performed by infusion method and four different concentrations of the drugs were encapsulated (200; 100; 50; 25 µg/ml). Free molecules of drugs were removed by dialysis. The encapsulation was verified using spectrophotometric and electrochemical methods. The amount of encapsulated drug linearly increased with the amount of applied drug (determination coefficient R2=0.8013). 76% of applied drug was encapsulated in phage λ particles (concentration of 10 µg/ml), even with the highest applied concentration of drugs, 200 µg/ml. Only 1% of encapsulated drug was detected in phage DNA. Similar results were obtained with encapsulation in phage empty capsid. Therefore, it can be concluded that the encapsulation of drugs into phage particles is efficient and mostly occurs by interaction of drugs with protein capsid.

Keywords: cytostatics, drug delivery, nanocarriers, phage capsid

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

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

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

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Authors: A. Hamieh, Z. Olama, H. Holail

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Drinking Water Distribution Systems provide opportunities for microorganisms that enter the drinking water to develop into biofilms. Antimicrobial agents, mainly chlorine, are used to disinfect drinking water, however, there are not yet standardized disinfection strategies with reliable efficacy and development of novel anti-biofilm strategies is still of major concern. In the present study the ability of Lactobacillus acidophilus and Streptomyces sp. cell free supernatants to inhibit the bacterial biofilm formation in Drinking Water Distribution System in Lebanon was investigated. Treatment with cell free supernatants of Lactobacillus acidophilus and Streptomyces sp. at 20% concentration resulted in average biofilm inhibition (52.89 and 39.66% respectively). A preliminary investigation about the mode of action of biofilm inhibition revealed that cell free supernatants showed no bacteriostatic or bactericidal activity against all the tested isolates. Pre-coating wells with supernatants revealed that Lactobacillus acidophilus cell free supernatant inhibited average biofilm formation (62.53%) by altering the adhesion of bacterial isolates to the surface, preventing the initial attachment step, which is important for biofilm production.

Keywords: biofilm, cell free supernatant, distribution system, drinking water, lactobacillus acidophilus, streptomyces sp, adhesion

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Authors: Anna Tokajuk, Agnieszka Zakrzeska, Ewa Chabielska, Halina Car

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Whey is a by-product generated mainly in the production of cheese and casein. Powder forms of whey are used widely in the food industry as well as a high-protein food for infants, for convalescents, by athletes and especially by bodybuilders to increase muscle mass during exercise. Whey protein concentrate-80 (WPC-80) is a source of bioactive peptides with beneficial effects on the cardiovascular system. It is known that whey proteins health beneficial properties include antidiabetic, blood pressure lowering, improving cardiovascular system function, antibacterial, antiviral and other effects. To study its influence on the development of thrombosis, venous thrombosis model was performed according to the protocol featured by Reyers with modification by Chabielska and Gromotowicz. Male Wistar-Crl: WI (Han) rats from researched groups were supplemented with two doses of WPC-80 (0.3 or 0.5 g/kg) for 7, 14 or 21 days and after these periods, one-hour venous thrombosis model was performed. Control group received 0.9 % NaCl solution and was sham operated. The statistical significance of results was computed by Mann – Whitney’s test. We observed that thrombus weight was decreased in animals obtaining WPC-8080 and that was statistically significant in 14 and 21-day supplemented groups. Blood count parameters did not differ significantly in rats with and without thrombosis induction whether they were fed with WPC-80 or not. Moreover, the number of platelets (PLT) was within the normal range in each group. The examined coagulation parameters in rats of the control groups were within normal limits. After WPC-80 supplementation there was the tendency to prolonged activated partial thromboplastin time (aPTT), but in comparison, the results did not turn out significant. In animals that received WPC-80 0.3 g·kg-1 for 21 days with and without induced thrombosis, prothrombin time (PT) and an international normalized ratio (INR) was somewhat decreased, remaining within the normal range, but the nature and significance of this observation are beyond the framework of the current study. Additionally, fibrinogen and thrombin time (TT) did not differ significantly between groups. Therefore the exact effect of WPC-80 on coagulation system is still elusive and requires further thorough research including mechanisms of action. Determining the potential clinical application of WPC-80 requires the selection of the optimal dose and duration of supplementation.

Keywords: antithrombotic, rats, venous thrombosis, WPC-80

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Authors: Sabah El-Sawalhi, Elie Fayad, Roula M. Abdel-Massih

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Ilex paraguariensis (Yerba Mate) is a medium to large tree commonly consumed by South Americans. Its leaves and stems are associated with different biological activities. The purpose of this study was to evaluate the antibacterial activity of Yerba Mate against Gram-positive and Gram-negative bacterial strains and its action against some resistant bacteria with different resistance profiles. Yerba Mate aqueous extracts were prepared at 70°C for 2 hrs, and the microdilution method was used to determine the minimum inhibitory concentration (MIC). Gram-positive bacteria exhibited a stronger antibacterial activity (MIC ranged between 0.468 mg/mL and 15 mg/mL) than Gram-negative bacteria. Yerba Mate was also extracted with acetone: water (1:1) and then further sub-fractionated with hexane, chloroform, and ethyl acetate. MIC values against Staphylococcus aureus ranged from 0.78 to 2.5 mg/ml for the chloroform fraction, from 1.56 to 3.75 mg/ml for the ethyl acetate fraction, and 0.78 to 1.87 mg/ml for the water fraction. The water fraction also exhibited antibacterial activity against Salmonella species (MIC ranged from 1.56 mg/ml to 3.12 mg/ml). The water fraction exhibited the highest antibacterial activity among all the fractions obtained. More studies are needed to determine the molecule or molecules responsible for this activity.

Keywords: antibacterial activity, bacterial resistance, minimum inhibitory concentration, yerba mate

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

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

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

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Authors: Elif Tugce Aksun Tumerkan, Umran Cansu, Gokhan Boran, Fatih Ozogul

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Gelatin is a mixture of peptides obtained from collagen by partial thermal hydrolysis. It is an important and useful biopolymer that is used in the food, pharmacy, and photography products. Generally, gelatins are sourced from pig skin and bones, beef bone and hide, but within the last decade, using alternative gelatin resources has attracted some interest. In this study, functional properties of gelatin extracted from seafood and poultry by-products were evaluated. For this purpose, skins of skipjack tuna (Katsuwonus pelamis) and frog (Rana esculata) were used as seafood by-products and chicken skin as poultry by-product as raw material for gelatin extraction. Following the extraction of gelatin, all samples were lyophilized and stored in plastic bags at room temperature. For comparing gelatins obtained; chemical composition, common quality parameters including bloom value, gel strength, and viscosity in addition to some others like melting and gelling temperatures, hydroxyproline content, and colorimetric parameters were determined. The results showed that the highest protein content obtained in frog gelatin with 90.1% and the highest hydroxyproline content was in chicken gelatin with 7.6% value. Frog gelatin showed a significantly higher (P < 0.05) melting point (42.7°C) compared to that of fish (29.7°C) and chicken (29.7°C) gelatins. The bloom value of gelatin from frog skin was found higher (363 g) than chicken and fish gelatins (352 and 336 g, respectively) (P < 0.05). While fish gelatin had higher lightness (L*) value (92.64) compared to chicken and frog gelatins, redness/greenness (a*) value was significantly higher in frog skin gelatin. Based on the results obtained, it can be concluded that skins of different animals with high commercial value may be utilized as alternative sources to produce gelatin with high yield and desirable functional properties. Functional and quality analysis of gelatin from frog, chicken, and tuna skin showed by-product of poultry and seafood can be used as an alternative gelatine source to mammalian gelatine. The functional properties, including bloom strength, melting points, and viscosity of gelatin from frog skin were more admirable than that of the chicken and tuna skin. Among gelatin groups, significant characteristic differences such as gel strength and physicochemical properties were observed based on not only raw material but also the extraction method.

Keywords: chicken skin, fish skin, food industry, frog skin, gel strength

Procedia PDF Downloads 159

Authors: M. Doudi, M. H. Pazandeh, L. Rahimzadeh Torabi

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Bedsores are pressure ulcers that occur on the skin or tissue due to being immobile and lying in bed for extended periods. Bedsores have the potential to progress into open ulcers, increasing the possibility of a variety of bacterial infections. Acinetobacter baumannii, a pathogen of considerable clinical importance, exhibited a significant correlation with Bedsores (pressure ulcers) infections, thereby manifesting a wide spectrum of antibiotic resistance. The emergence of drug resistance has led researchers to focus on alternative methods, particularly phage therapy, for tackling bacterial infections. Phage therapy has emerged as a novel therapeutic approach to regulate the activity of these agents. The management of bacterial infections greatly benefits from the clinical utilization of bacteriophages as a valuable antimicrobial intervention. The primary objective of this investigation consisted of isolating and discerning potent bacteriophage capable of targeting multi-drug-resistant (MDR) and extensively drug-resistant (XDR) bacteria obtained from pressure ulcers. The present study analyzed and isolated A. baumannii strains obtained from a cohort of patients suffering from pressure ulcers at Taleghani Hospital in Ahvaz, Iran. An approach that included biochemical and molecular identification techniques was used to determine the taxonomic classification of bacterial isolates at the genus and species levels. The molecular identification process was facilitated by using the 16S rRNA gene in combination with universal primers 27 F and 1492 R. Bacteriophage was obtained through the isolation process conducted on treatment plant sewage located in Isfahan, Iran. The main goal of this study was to evaluate different characteristics of phage, such as their appearance, the range of hosts they can infect, how quickly they can enter a host, their stability at varying temperatures and pH levels, their effectiveness in killing bacteria, the growth pattern of a single phage stage, mapping of enzymatic digestion, and identification of proteomics patterns. The findings demonstrated that an examination was conducted on a sample of 50 specimens, wherein 15 instances of A. baumannii were identified. These microorganisms are the predominant Gram-negative agents known to cause wound infections in individuals suffering from bedsores. The study's findings indicated a high prevalence of antibiotic resistance in the strains isolated from pressure ulcers, excluding the clinical strains that exhibited responsiveness to colistin. According to the findings obtained from assessments of host range and morphological characteristics of bacteriophage VbɸAB-1, it can be concluded that this phage possesses specificity towards A. Baumannii BAH_Glau1001 was classified as a member of the Podoviridae family. The bacteriophage mentioned earlier showed the strongest antibacterial effect at a temperature of 18 °C and a pH of 6.5. Through the utilization of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis on protein fragments, it was established that the bacteriophage VbɸAB-1 exhibited a size range between 50 and 75 kilodaltons (KDa). The numerous research findings on the effectiveness of phages and the safety studies conducted suggest that the phages studied in this research can be considered as a practical solution and recommended approach for controlling and treating stubborn pathogens in burn wounds among hospitalized patients. The findings of our research indicated that isolated phages could be an effective antimicrobial and an appreciate candidate for prophylaxis against pressure ulcers.

Keywords: acinetobacter baumannii, extremely drug-resistant, phage therapy, surgery wound

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