Search results for: origanum glandulosum

Authors: A. Lardjam, R. Mazid, S. Y. Boudghene, A. Izarouken, Y. Dali, N. Djebli, H. Toumi

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Origanum glandulosum is an aromatic plant, common in Algeria and widely used by local people for its medicinal properties. The essential oil from this plant, which grows in the west of Algeria, was studied to evaluate and determine its antibacterial activity. The extraction of the essential oil was performed by water steam distillation; the yield obtained from the aerial parts (1.78 %) is interesting, its chromatographic profile revealed by TLC showed the presence of phenolic compounds thymol and carvacrol. The evaluation of the activity of the essential oil of Origanum glandulosum on bacterial strains of hospital origin, ATCC, MRB, and HRB, most implicated in nosocomial infections (Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 43300, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus resistant to meticillin, Enterococcus faecium, VA R and R TEC, Acinetobacter baumanii, IMP R and R CAZ, Klebsiella pneumonia carbapenemase-producing) by the method of aromatogramme and micro atmosphere, shows that the antibacterial potency of this oil is very high, expressed by significant inhibition diameters on all strains except Pseudomonas aeruginosa, and low MICs and is characterized by a bactericidal action.

Keywords: antibacterial activity, essential oil, HRB, MBR, nosocomial infections, origanum glandulosum

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Authors: Abdelkader Basli, Jean-Claude Delaunay, Eric Pedrot, Jean-Michel Mérillon, Jean-Pierre Monti, Khodir Madani, Mohamed Chibane, Tristan Richard

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The antioxidant and antimicrobial activities of Origanum glandulosum collected in Algeria have been studied. Extract was prepared from aerial part of endemic Algerian oregano. The produced extract has been characterized in terms of total phenols (using Folin method), total flavonoid, antioxidant activities (using the DPPH radical scavenging method and ORAC assay) and microbial activity against four bacteria: Streptococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae one yeast: Candida albicans and one fungi: Aspergillus niger. The results pointed the antioxidant activities of the extract of O. glandulosum and antimicrobial activities against all bacteria and C. Candida, but no effect on A. niger. High performance liquid chromatography combined with mass spectrometry (LC-MS) and nuclear magnetic resonance (LC-NMR) were used to separate and identify the major compounds present in the oregano extract. Rosmarinic acid, globoidnan A and B, lithospermic acid B and three flavonoids were identified.

Keywords: origanum glandulosum, antioxidant, microbial activity, polyphenol, LC-MS, LC-NMR

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Authors: S. Krimat, T. Dob, L. Lamari, H. Metidji

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The aim of this study is to evaluate the antioxidant and antimicrobial activity of hydromethanolic extract of selected Algerian medicinal flora. The antioxidant activity of extract was evaluated in terms of radical scavenging potential (DPPH) and β-carotene bleaching assay. Total phenolic contents and flavonoid contents were also measured. Antimicrobial activity of these plants was tested against five microorganisms Pseu-domonas aeruginosa Bacillus subtilis, Escherichia coli, Staphylococcus aureus, and Candida albicans. The results showed that Pistacia lentiscus showed the highest antioxidant capacities using DPPH assay (IC50 = 4.60 μg/ml), while Populus trimula had the highest antioxidant activity in β-carotene/linolaic acid assay. The most interesting antimicrobial activity was obtained from Sysimbrium officinalis, Rhamnus alaternus, Origanum glandulosum, Cupressus sempervirens, Pinus halipensis and Centaurea calcitrapa. The results indicate that the plants tested may be potential sources for isolation of natural antioxidant and antimicrobial compounds.

Keywords: Algerian medicinal plants, antimicrobial activity, antioxidant activity, disc diffusion method

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Authors: A.Assariotakis, P. Vahamidis, P. Tarantilis, G. Economou

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Greece, due to its geographical location and the particular climatic conditions, presents high biodiversity of Medicinal and Aromatic Plants. Among them, the genus Origanum not only presents a wide distribution, but it also has great economic importance. After extensive surveys in Ikaria Island (Greece), 3 species of the genus Origanum were identified, namely, Origanum vulgare ssp. hirtum (Greek oregano), Origanum onites (Turkish oregano) and Origanum x intercedens (hybrid), a naturally occurring hybrid between O. hirtum and O. onites. The purpose of this study was to determine their morphological as well as their temporal variability in essential oil yield and composition under field conditions. For this reason, a plantation of each species was created using vegetative propagation and was established at the experimental field of the Agricultural University of Athens (A.U.A.). From the establishment year and for the following two years (3 years of observations), several observations were taken during each growing season with the purpose of identifying the morphological differences among the studied species. Each year collected plant (at bloom stage) material was air-dried at room temperature in the shade. The essential oil content was determined by hydrodistillation using a Clevenger-type apparatus. The chemical composition of essential oils was investigated by Gas Chromatography-Mass Spectrometry (GC – MS). Significant differences were observed among the three oregano species in terms of plant height, leaf size, inflorescence features, as well as concerning their biological cycle. O. intercedens inflorescence presented more similarities with O. hirtum than with O. onites. It was found that calyx morphology could serve as a clear distinction feature between O. intercedens and O. hirtum. The calyx in O. hirtum presents five isometric teeth whereas in O. intercedens two high and three shorter. Essential oil content was significantly affected by genotype and year. O. hirtum presented higher essential oil content than the other two species during the first year of cultivation, however during the second year the hybrid (O. intercedens) recorded the highest values. Carvacrol, p-cymene and γ-terpinene were the main essential oil constituents of the three studied species. In O. hirtum carvacrol content varied from 84,28 - 93,35%, in O. onites from 86,97 - 91,89%, whereas in O. intercedens it was recorded the highest carvacrol content, namely from 89,25 - 97,23%.

Keywords: variability, oregano biotypes, essential oil, carvacrol

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Authors: Eva Tvrdá, Barbora Babečková, Michal Ďuračka, Róbert Kirchner, Július Árvay

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This study was designed to assess the in vitro effects of Origanum vulgare L. (oregano) extract on the testicular steroidogenesis. We focused on identifying major biomolecules present in the oregano extract, as well as to investigate its in vitro impact on the secretion of cholesterol, testosterone, dehydroepiandrosterone and androstenedione by murine testicular fragments. The extract was subjected to high performance liquid chromatography (HPLC) which identified cyranosid, daidzein, thymol, rosmarinic and trans-caffeic acid among the predominant biochemical components of oregano. For the in vitro experiments, testicular fragments from 20 sexually mature Institute of Cancer Research (ICR) mice were incubated in the absence (control group) or presence of the oregano extract at selected concentrations (10, 100 and 1000 μg/mL) for 24 h. Cholesterol levels were quantified using photometry and the hormones were assessed by ELISA (Enzyme-Linked Immunosorbent Assay). Our data revealed that the release of cholesterol and androstenedione (but not dehydroepiandrosterone and testosterone) by the testicular fragments was significantly impacted by the oregano extract in a dose-dependent fashion. Supplementation of the extract resulted in a significant decline of cholesterol (P < 0.05 in case of 100 μg/mL; P < 0.01 with respect 100 μg/mL extract), as well as androstenedione (P < 0.01 with respect to 100 and 1000 μg/mL extract). Our results suggest that the biomolecules present in Origanum vulgare L. could exhibit a dose-dependent impact on the secretion of male steroids, playing a role in the regulation of testicular steroidogenesis.

Keywords: mice, Origanum vulgare L., steroidogenesis, testes

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Authors: Ashkan Zargar, Ali Taheri Mirghaed, Zein Talal Barakat, Alireza Khosravi, Hamed Paknejad

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With the increase of bacterial resistance to the chemical antibiotics, replacing it with ecofriendly herbal materials and with no adverse effects in the host body is very important. Therefore, in this study, the effect of combined essential oil (Thymus vulgaris-Origanum magorana and Ziziphora clinopodioides) on the growth behavior of Yersinia ruckeri, Aeromonas hydrophila and Lactococcus garvieae was evaluated. The compositions of the herbal essential oils used in this study were determined by gas chromatography-mass spectrometry (GC-MS) while, the investigating of antimicrobial effects was conducted by the agar-disc diffusion method, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and bacterial growth curves determination relied on optical density (OD) at 630 nm. The main compounds were thymol (40.60 %) and limonene (15.98 %) for Thymus vulgaris while carvacrol (57.86 %) and thymol (13.54 %) were the major compounds in Origanum magorana. As regards Ziziphora clinopodiodes, α-pinene (22.6 %) and carvacrol (21.1 %) represented the major constituents. Concerning Yersinia ruckeri, disc-diffusion results showed that t.O.z (50 % Origanum majorana) combined essential oil was presented the best inhibition zone (30.66 mm) but it was exhibited no significant differences with other tested commercial antibiotics except oxytetracycline (P <0/05). The inhibitory activity and the bactericidal effect of the t.O.z, unveiled by the MIC= 0.2 μL /mL and MBC= 1.6 μL /mL values, were clearly the best between all combined oils. The growth behaviour of Yersinia ruckeri was affected by this combined essential oil and changes in temperature and pH conditions affected herbal oil performance. As regard Aeromonas hydrophila, its results were so similar to Yersinia ruckeri results and t.O.z (50 % Origanum majorana) was the best between all combined oils (inhibition zone= 26 mm, MIC= 0.4 μL /mL and MBC= 3.2 μL /mL, combined essential oil was affected bacterial growth behavior). Also for Lactococcus garvieae, t.O.z (50 % Origanum majorana) was the best between all combined oils having the best inhibition zone= 20.66 mm, MIC= 0.8 μL /mL and MBC= 1.6 μL /mL and best effect on inhibiting bacterial growth. Combined herbal essential oils have a good and noticeable effect on the growth behavior of pathogenic bacteria in the laboratory, and by continuing research in the host, they may be a suitable alternative to control, prevent and treat diseases caused by these bacteria.

Keywords: bacterial pathogen, herbal medicine, growth behavior, fish

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Authors: Fatih Sonmez

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Alzheimer’s disease (AD) is a progressive neurodegenerative disease and it is the most common form of dementia that affects aged people. Acetylcholinesterase is a hydrolase involved in the termination of impulse transmission at cholinergic synapses by rapid hydrolysis of the neurotransmitter ACh in the central and peripheral nervous system. Carvacrol (5-iso-propyl-2-methyl-phenol) is a main bioactive monoterpene isolated from many medicinal herbs, such as Thymus vulgaris, Monarda punctate and Origanum vulgare spp. It is known that carvacrol has been widely used as an active anti-inflammatory ingredient, which can inhibit the isoproterenol induced inflammation in myocardial infarcted rats. In this paper, a series of 12 carvacrol substituted carbamate derivatives (2a-l) was synthesized and their inhibitory activities on AChE and BuChE were evaluated. Among them, 2d exhibited the strongest inhibition against AChE with an IC50 value of 2.22 µM, which was 130-fold more than that of carvacrol (IC50 = 288.26 µM).

Keywords: Acetylcholinesterase, Butyrylcholinesterase, Carbamate, Carvacrol

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Authors: Ivana Cabarkapa, Zorica Tomicic, Olivera Duragic

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Antimicrobial resistance represents one of the major challenges facing humanity in the last decades. Increasing antibiotic-resistant pathogens indicates the need for the development of alternative antibacterial drugs and new treatment strategies. One of the innovative emerging treatments in overcoming multidrug-resistant pathogens certainly represents the inhibition anti-quorum sensing system. For most of the food-borne pathogens, the expression of the virulence depends on their capability communication with other members of the population by means of quorum sensing (QS). QS represents a specific way of bacterial intercellular communication, which enabled owing to their ability to detect and to respond to cell population density by gene regulation. QS mechanisms are responsible for controls the pathogenesis, virulence luminescence, motility, sporulation and biofilm formation of many organisms by regulating gene expression. Therefore, research in this field is being an attractive target for the development of new natural antibacterial agents. Anti-QS compounds are known to have the ability to prohibit bacterial pathogenicity. Considering the importance of quorum sensing during bacterial pathogenesis, this research has been focused on evaluation anti - QS properties of four essential oils (EOs) Origanum heracleoticum, Origanum vulgare, Thymus vulgare, and Thymus serpyllum, using biomonitor strain of Chromobacterium violaceum CV026. Tests conducted on Luria Bertani agar supplemented with N hexanol DL homoserine lacton (HHL) 10µl/50ml of agar. The anti-QS potential of the EOs was assayed in a range of concentrations of 200 – 0.39 µl/ml using the disc diffusion method. EOs of Th. vulgaris and T. serpyllum were exhibited anti-QS activity indicated by a non- pigmented ring with a dilution-dependent manner. The lowest dilution of EOs T. vulgaris and T. serpyllum in which they exhibited visually detectable inhibition of violacein synthesis was 6.25 µl/ml for both tested EOs. EOs of O. heracleoticum and O. vulgare were displayed different active principles, i.e., antimicrobial activity indicated by the inner clear ring and anti-QS activity indicated by the outer non-pigmented ring, in a concentration-dependent manner. The lowest dilution of EOs of O. heracleoticum and O. vulgare in which exhibited visually detectable inhibition of violacein synthesis was 1.56 and 3.25 µl/ml, respectively. Considering that, the main constituents of the tested EOs represented by monoterpenes (carvacrol, thymol, γ-terpinene, and p-cymene), anti - QS properties of tested EOs can be mainly attributed to their activity. In particular, from the scientific literature, carvacrol and thymol show a sub-inhibitory effect against foodborne pathogens. Previous studies indicated that sub-lethal concentrations of carvacrol reduced the mobility of bacteria due to the ability of interference using QS mechanism between the bacterial cells, and thereby reducing the ability of biofilm formation The precise mechanism by which carvacrol inhibits biofilm formation is still not fully understood. Our results indicated that EOs displayed different active principles, i.e., antimicrobial activity indicated by the inner clear ring and anti-QS activity indicated by an outer non- pigmented ring with visually detectable inhibition of violacein. Preliminary results suggest that EOs represent a promising alternative for effective control of the emergence and spread of resistant pathogens.

Keywords: anti-quorum sensing activity, Chromobacterium violaceum, essential oils, violacein

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Authors: Anita Vidacs, Csaba Vagvolgyi, Judit Krisch

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The attachment of microorganisms to different surfaces and the development of biofilms can lead to outbreaks of food-borne diseases and economic losses due to perished food. In food processing environments, bacterial communities are generally formed by mixed cultures of different species. Plants are sources of several antimicrobial substances that may be potential candidates for the development of new disinfectants. We aimed to investigate cinnamon (Cinnamomum zeylanicum), marjoram (Origanum majorana), and thyme (Thymus vulgaris). Essential oils and their major components (cinnamaldehyde, terpinene-4-ol, and thymol) on four-species biofilms of E. coli, L. monocytogenes, P. putida, and S. aureus. Experiments had three parts: (i) determination of minimum bactericide concentration and the killing time with microdilution methods; (ii) elimination of the four-species 24– and 168-hours old biofilm from stainless steel, polypropylene, tile and wood surfaces; and (iii) comparing the disinfectant effect with industrial used per-acetic based sanitizer (HC-DPE). E. coli and P. putida were more resistant to investigated essential oils and their main components in biofilm, than L. monocytogenes and S. aureus. These Gram-negative bacteria were detected on the surfaces, where the natural based disinfectant had not total biofilm elimination effect. Most promoted solutions were the cinnamon essential oil and the terpinene-4-ol that could eradicate the biofilm from stainless steel, polypropylene and even from tile, too. They have a better disinfectant effect than HC-DPE. These natural agents can be used as alternative solutions in the battle against bacterial biofilms.

Keywords: biofilm, essential oils, surfaces, terpinene-4-ol

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Authors: Anil F. Felek, Mehmet M. Ozcan, Faruk Akyazi

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Volatile oils of medicinal and aromatic plants are important for managing nematological problems in agriculture. In present study, volatile oils extracted from five medicinal and aromatic plants including Origanum onites (flower+steam+leaf), Salvia officinalis (leaf), Lippia citriodora (leaf+seed), Mentha spicata (leaf) and Mentha longifolia (leaf) were tested for egg hatching inhibition activity against root-knot nematode Meloidogyne hapla under laboratory conditions. The essential oils were extracted using water distillation method with a Clevenger system. For the homogenisation process of the oils, 2% gum arabic solution was used and 4 µl oils was added into 1ml filtered gum arabic solution to prepare the last stock solution. 5 ml of stock solution and 1 ml of M. hapla egg suspension (about 100 eggs) were added into petri dishes. Gum arabic solution was used as control. Seven days after exposure to oils at room temperature (26±2 °C), the cumulative hatched and unhatched eggs were counted under 40X inverted light microscope and Abbott’s formula was used to calculate egg hatching inhibition rates. As a result, the highest inhibition rate was found as 54% for O. onites. In addition, the other inhibition rates varied as 31.4%, 21.6%, 23.8%, 25.67% for the other plants, S. officinalis, M. longifolia, M. spicata and L. citriodora, respectively. Carvacrol was found as the main component (68.8%) of O. onites followed by Thujone 27.77% for S. officinalis, I-Menthone 76.92% for M. longifolia, Carvone 27.05% for M. spicata and Citral 19.32% for L. citriodora.

Keywords: egg hatching, Meloidogyne hapla, medicinal and aromatic plants, root-knot nematodes, volatile oils

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Authors: Tomas Drevinskas, Ruta Mickiene, Audrius Maruska, Nicola Tiso, Algirdas Salomskas, Raimundas Lelesius, Agneta Karpovaite, Ona Ragazinskiene, Loreta Kubiliene

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Antiviral effect of substances accumulated by plants and natural products is known to ethno-pharmacy and modern day medicine. Antiviral properties are usually assigned to volatile compounds and polyphenols. This research work is divided into several parts and the task of this part was to investigate potential plants, potential substances and potential preparation conditions that can be used for the preparation of antiviral agents. Sixteen different medicinal plants, their parts and two types of propolis were selected for screening. Firstly, extraction conditions of non-volatile compounds were investigated: 3 pre-selected plants were extracted with 5 different ethanol – water mixtures (96%, 75%, 60%, 40%, 20 %, vol.) and bidistilled water. Total phenolic content, total flavonoid content and radical scavenging activity was determined. The results indicated that optimal extrahent is 40%, vol. of ethanol – water mixture. Further investigations were performed with the extrahent of 40%, vol. ethanol – water mixture. All 16 of selected plants, their parts and two types of propolis were extracted using selected extrahent. Determined total phenolic content, total flavonoid content and radical scavenging activity indicated that extracts of Origanum Vulgare L., Mentha piperita L., Geranium macrorrhizum L., Melissa officinalis L. and Desmodium canadence L. contains highest amount of extractable phenolic compounds (7.31, 5.48, 7.88, 8.02 and 7.16 rutin equivalents (mg/ ml) respectively), flavonoid content (2.14, 2.23, 2.49, 0.79 and 1.51 rutin equivalents (mg/ml) respectively) and radical scavenging activity (11.98, 8.72, 13.47, 13.22 and 12.22 rutin equivalents (mg/ml) respectively). Composition of the extracts is analyzed using HPLC.

Keywords: antiviral effect, plants, propolis, phenols

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Authors: Akbar Zeb, Shujaul Mulk Khan, Habib Ahmad, Manzoor Hussain, Mujtaba Shah

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An ethnobotanical project was carried out in the Dughalgay valley District Swat in Hindu Kush region. The Local population not only use indigenous knowledge to use medicinal plants for curing various diseases but also earn their live hood by selling some of them in the local markets. An ethnobotanical project was carried out in the Doghalgay valley of upper Swat. The Local population not only use indigenous medicinal plants for curing various diseases but also earn their live hood by selling some of them in the local market. 102 of these medicinal plants were reported to be used in the region during questionnaire survey in spring 2007. Out of them 10 species are used as diuretic, 9 in stomachic and laxative each. Similarly 6, 5, 5, 4, 4, and 4 species of them are used as antiseptic, Anthelmintic, Carminative, Expectorant, Astringent and purgative respectively, while the remaining species have one or more than one medicinal use in the local community. 30 of these species are collected for marketing purposes, in which these medicinal plants such as Berberis lycium, Origanum vulgare, Bergenia ciliata, Aesculus indica, Podophyllum emodi, Pteredium aquilinum, Bergenia himalyca, Viola spp., Ajuga bracteosa, Morchella esculenta, Paeonia emodi, Atropa acuminate, Aconitum violaceum, Polygonum amplexicaulis, Bupleurum longicaule, Juglans regia, Diospyrus lotus, and Mentha longifolia are important. Study concluded that availability of medicinal plants is decreasing day by day due to human population pressure, marketing pressure, grazing and unwise collection. Therefore it is recommended that Governmental organizations and non Governmental organization should pay possible attention to make aware the local people about the future threats.

Keywords: indigenous knowledge, ethnomedicinal uses, marketing, Hindu Kush

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Authors: Cristian Moisa, Andreea Lupitu, Adriana Csakvari, Dana G. Radu, Leonard Marian Olariu, Georgeta Pop, Dorina Chambre, Lucian Copolovici, Dana Copolovici

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Green synthesis of nanoparticles (NPs) represents a promising, accessible, eco-friendly, and safe process with significant applications in biotechnology, pharmaceutical sciences, and farming. The aim of our study was to obtain silver nanoparticles, using plant wastes extracts resulted in the essential oils extraction process: Thymus vulgaris L., Origanum vulgare L., Lavandula angustifolia L., and in hemp processing for seed and fibre, Cannabis sativa. Firstly, we obtained aqueous extracts of thyme, oregano, lavender, and hemp (two monoicous and one dioicous varieties), all harvested in western part of Romania. Then, we determined the chemical composition of the extracts by liquid-chromatography coupled with diode array and mass spectrometer detectors. The compounds identified in the extracts were in agreement with earlier published data, and the determination of the antioxidant activity of the obtained extracts by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assays confirmed their antioxidant activity due to their total polyphenolic content evaluated by Folin-Ciocalteu assay. Then, the silver nanoparticles (AgNPs) were successfully biosynthesised, as was demonstrated by UV-VIS, FT-IR spectroscopies, and SEM, by reacting AgNO₃ solution and plant extracts. AgNPs were spherical in shape, with less than 30 nm in diameter, and had a good bactericidal activity against Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas fluorescens).

Keywords: plant wastes extracts, chemical composition, high performance liquid chromatography mass spectrometer, HPLC-MS, scanning electron microscopy, SEM, silver nanoparticles

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Authors: Sumira Jan, Javid Iqbal Mir, Desh Beer Singh, Anil Sharma, Shafia Zaffar Faktoo

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Background and Aims: Climatic and edaphic factors immensely influence crop quality and proper development. Regardless of economic potential, Himalayan Oregano has not subjected to phytonutrient and chemometric evaluation and its relationship with environmental conditions are scarce. The central objective of this research was to investigate microclimatic variation among wild and cultivated populations located in a microclimatic gradient in north-western Himalaya, Kashmir and analyse if such disparity was related with diverse climatic and edaphic conditions. Methods: Micrometeorological, Atomic absorption spectroscopy for micro elemental analysis was carried for soil. HPLC was carried out to estimate variation in phytonutrients and phytochemicals. Results: Geographic variation in phytonutrient was observed among cultivated and wild populations and among populations diverse within regions. Cultivated populations exhibited comparatively lesser phytonutrient value than wild populations. Moreover, our results observed higher vegetative growth of O. vulgare L. with higher pH (6-7), elevated organic carbon (2.42%), high nitrogen (97.41Kg/ha) and manganese (10-12ppm) and zinc contents (0.39-0.50) produce higher phytonutrients. HPLC data of phytonutrients like quercetin, betacarotene, ascorbic acid, arbutin and catechin revealed direct relationship with UV-B flux (r2=0.82), potassium (r2=0.97) displaying parallel relationship with phytonutrient value. Conclusions: Catechin was found as predominant phytonutrient among all populations with maximum accumulation of 163.8 ppm while as quercetin exhibited lesser value. Maximum arbutin (53.42ppm) and quercetin (2.87ppm) accumulated in plants thriving under intense and high UV-B flux. Minimum variation was demonstrated by beta carotene and ascorbic acid.

Keywords: phytonutrient, ascorbic acid, beta carotene, quercetin, catechin

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Authors: Kubilay Kurtulus Bastas

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Bacterial diseases are very destructive and cause economic losses on pears. Promising plant extracts for the management of plant diseases are environmentally safe, long-lasting and extracts of certain plants contain alkaloids, tannins, quinones, coumarins, phenolic compounds, and phytoalexins. In this study, bacteria were isolated from different parts of pear exhibiting characteristic symptoms of bacterial diseases from the Central Anatolia, Turkey. Pathogenic bacteria were identified by morphological, physiological, biochemical and molecular methods as fire blight (Erwinia amylovora (39%)), bacterial blossom blast and blister bark (Pseudomonas syringae pv. syringae (22%)), crown gall (Rhizobium radiobacter (1%)) from different pear cultivars, and determined virulence levels of the pathogens with pathogenicity tests. The air-dried 25 plant material was ground into fine powder and extraction was performed at room temperature by maceration with 80% (v/v) methanol/distilled water. The minimum inhibitory concentration (MIC) values were determined by using modified disc diffusion method at five different concentrations and streptomycin sulphate was used as control chemical. Bacterial suspensions were prepared as 108 CFU ml⁻¹ densities and 100 µl bacterial suspensions were spread to TSA medium. Antimicrobial activity was evaluated by measuring the inhibition zones in reference to the test organisms. Among the tested plants, Origanum vulgare, Hedera helix, Satureja hortensis, Rhus coriaria, Eucalyptus globulus, Rosmarinus officinalis, Ocimum basilicum, Salvia officinalis, Cuminum cyminum and Thymus vulgaris showed a good antibacterial activity and they inhibited the growth of the pathogens with inhibition zone diameter ranging from 7 to 27 mm at 20% (w/v) in absolute methanol in vitro conditions. In vivo, the highest efficacy was determined as 27% on reducing tumor formation of R. radiobacter, and 48% and 41% on reducing shoot blight of E. amylovora and P. s. pv. syringae on pear seedlings, respectively. Obtaining data indicated that some plant extracts may be used against the bacterial diseases on pome fruits within sustainable and organic management programs.

Keywords: bacteria, eco-friendly management, organic, pear, plant extract

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Authors: Valeria Velasco, Ana M. Bonilla, José L. Vergara, Alcides Lofa, Jorge Campos, Pedro Rojas-García

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Staphylococcus aureus is both commensal bacterium and opportunistic pathogen that can cause different diseases in humans and can rapidly develop antimicrobial resistance. Since this bacterium has the ability to colonize the nares and skin of humans and animals, there is a risk of contamination of food in different steps of the food chain supply. Emerging strains have been detected in food-producing animals and meat, such as methicillin-resistant S. aureus (MRSA). The aim of this study was to determine the prevalence and oxacillin susceptibility of S. aureus in the pork chain supply in Chile and to suggest some natural antimicrobials for control. A total of 487 samples were collected from pigs (n=332), carcasses (n=85), and retail pork meat (n=70). Presumptive S. aureus colonies were isolated by selective enrichment and culture media. The confirmation was carried out by biochemical testing (Api® Staph) and molecular technique PCR (detection of nuc and mecA genes, associated with S. aureus and methicillin resistance, respectively). The oxacillin (β-lactam antibiotic that replaced methicillin) susceptibility was assessed by minimum inhibitory concentration (MIC) using the Epsilometer test (Etest). A preliminary assay was carried out to test thymol, carvacrol, oregano essential oil (Origanum vulgare L.), Maqui or Chilean wineberry extract (Aristotelia chilensis (Mol.) Stuntz) as anti-staphylococcal agents using the disc diffusion method at different concentrations. The overall prevalence of S. aureus in the pork chain supply reached 33.9%. A higher prevalence of S. aureus was determined in carcasses (56.5%) than in pigs (28.3%) and pork meat (32.9%) (P ≤ 0.05). The prevalence of S. aureus in pigs sampled at farms (40.6%) was higher than in pigs sampled at slaughterhouses (23.3%) (P ≤ 0.05). The contamination of no packaged meat with S. aureus (43.1%) was higher than in packaged meat (5.3%) (P ≤ 0.05). The mecA gene was not detected in S. aureus strains isolated in this study. Two S. aureus strains exhibited oxacillin resistance (MIC ≥ 4µg/mL). Anti-staphylococcal activity was detected in solutions of thymol, carvacrol, and oregano essential oil at all concentrations tested. No anti-staphylococcal activity was detected in Maqui extract. Finally, S. aureus is present in the pork chain supply in Chile. Although the mecA gene was not detected, oxacillin resistance was found in S. aureus and could be attributed to another resistance mechanism. Thymol, carvacrol, and oregano essential oil could be used as anti-staphylococcal agents at low concentrations. Research project Fondecyt No. 11140379.

Keywords: antimicrobials, mecA gen, nuc gen, oxacillin susceptibility, pork meat

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Authors: R. A. Abdel Rahman, A. E. Abdel Wahab, E. A. Goghneimy, H. F. Mohamed, E. M. Salama

Abstract:

Infectious diseases are a major cause of death worldwide. The treatment of infections continues to be problematic in modern time because of the severe side effects of some drugs and the growing resistance to antimicrobial agents. Hence, the search for newer, safer and more potent antimicrobials is a pressing need. Herbal medicines have received much attention as a source of new antibacterial drugs since they are considered time-tested and comparatively safe both for human use and the environment. In the present study, the antimicrobial activity of marjoram (Origanum majorana L.) essential oil on some gram positive and gram negative reference bacteria, as well as some hospital resistant microbes, was tested. Marjoram oil was extracted and the oil chemical constituents were identified using GC/MS analysis. Staphylococcus aureas ATCC 6923, Pseudomonus auregonosa ATCC 9027, Bacillus subtilis ATCC 6633, E. coli ATCC 8736 and two hospital resistant microbes isolates 16 and 21 were used. The two isolates were identified by biochemical tests and 16s rRNA as proteus spp. and Enterococcus facielus. The effect of different concentrations of essential oils on bacterial growth was tested using agar disk diffusion assay method to determine the minimum inhibitory concentrations and using micro dilution method to determine the minimum bactericidal concentrations. Marjoram oil was found to be effective against both reference and hospital resistance strains. Hospital strains were more resistant to marjoram oil than reference strains. P. auregonosa growth was completely inhibited at a low concentration of oil (4µl/ml). The other reference strains showed sensitivity to marjoram oil at concentrations ranged from 5 to 7µl/ml. The two hospital strains showed sensitivity at media containing 10 and 15µl/ml oil. The major components of oil were terpineol, cis-beta (23.5%), 1,6 – octadien –3-ol,3,7-dimethyl, 2 aminobenzoate (10.9%), alpha terpieol (8.6%) and linalool (6.3%). Scanning electron microscope (SEM) and transmission electron microscope (TEM) analysis were used to determine the difference between treated and untreated hospital strains. SEM results showed that treated cells were smaller in size than control cells. TEM data showed that cell lysis has occurred to treated cells. Treated cells have ruptured cell wall and appeared empty of cytoplasm compared to control cells which shown to be intact with normal volume of cytoplasm. The results indicated that marjoram oil has a positive antimicrobial effect on hospital resistance microbes. Natural crude extracts can be perfect resources for new antimicrobial drugs.

Keywords: antimicrobial activity, essential oil, hospital resistance microbes, marjoram

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Authors: Ana Maria Muț, Georgeta Coneac, Ioana Olariu, Ștefana Avram, Ioana Zinuca Pavel, Ionela Daliana Minda, Lavinia Vlaia, Cristina Adriana Dehelean, Corina Danciu

Abstract:

Oregano essential oil is obtained from different parts of the plant Origanum vulgare (fam. Lamiaceae) and carvacrol and thymol are primary components, widely recognized for their antimicrobial activity, as well as their antiviral and antifungal properties. Poloxamers are triblock copolymers (Pluronic®), formed of three non-ionic blocks with a hydrophobic polyoxypropylene central chain flanked by two polyoxyethylene hydrophilic chains. They are known for their biocompatibility, sensitivity to temperature changes (sol-to-gel transition of aqueous solution with temperature increase), but also for their amphiphilic and surface active nature determining the formation of micelles, useful for solubilization of different hydrophobic compounds such as the terpenes and terpenoids contained in essential oils. Thus, these polymers, listed in European and US Pharmacopoeia and approved by FDA, are widely used as solubilizers and gelling agents for various pharmaceutical preparations, including topical hydrogels. The aim of this study was to investigate the posibility of solubilizing oregano essential oil (OEO) in polymeric micelles using polyoxypropylene (PPO)-polyoxyethylene (PEO)-polyoxypropylene (PPO) triblock polymers to obtain semisolid systems suitable for topical application. A formulation screening was performed, using Pluronic® F-127 in concentration of 20%, Pluronic® L-31, Pluronic® L-61 and Pluronic® L-62 in concentration of 0.5%, 0.8% respectively 1% to obtain the polymeric micelles-based systems. Then, to each selected system, with or without 10% absolute ethanol, 5% or 8% OEO was added. The obtained transparent poloxamer-based hydrogels containing solubilized OEO were further evaluated for pH, rheological characteristics (flow behaviour, viscosity, consistency and spreadability), using consacrated techniques like potentiometric titration, stationary shear flow test, penetrometric method and parallel plate method. Also, in vitro release and permeation of carvacrol from the hydrogels was carried out, using vertical diffusion cells and synthetic hydrophilic membrane and porcine skin respectively. The pH values and rheological features of all tested formulations were in accordance with official requirements for semisolid cutaneous preparations. But, the formulation containing 0.8% Pluronic® L-31, 10% absolute ethanol, 8% OEO and water and the formulation with 1% Pluronic® L-31, 5% OEO and water, produced the highest cumulative amounts of carvacrol released/permeated through the membrane. The present study demonstrated that oregano essential oil can be successfully solubilized in the investigated poloxamer-based hydrogels. These systems can be further investigated as potential topical therapy for cutaneous papillomas. Funding: This research was funded by Project PN-III-P1-1.1-TE2019-0130, Contract number TE47, Romania.

Keywords: oregano essential oil, carvacrol, poloxamer, topical hydrogels

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