Search results for: Rosemarinus officinalis
Commenced in January 2007
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Edition: International
Paper Count: 54

Search results for: Rosemarinus officinalis

54 Insecticidal and Antifeedant Activity of Rosemary´s (Rosmarinus Officinalis L.) Different Extracts on Cotton Bollworm Helicoverpa Armigera Hubner

Authors: Monireh Movahedi

Abstract:

Considering undesirable effects of chemical insecticides on environment and human health, most studies focused on insecticidal effects of plant materials. Here, the insecticidal effects of methanol, ethylacetat and n-Hexan extracts of Rosmarinus officinalis L. on larval stage of the cotton bollworm were studied. From each extract, six concentrations, including 5, 10, 20, 40 and 60 mg/ml were prepared and added on larval artificial diet. Moreover, solution of distilled water and tween 2% considered as check treatment. All experiments were done in laboratory temperature of 25±3 ºC, RH =50±10% and natural photoperiod during growing season. Each treatment had four replications and each replication carried out on 10 first instar larva with <24h age. Larval mortality was recorded 3 and 7 days after treat. Based on results, LC50 of methanol, ethylacetat and n-Hexan extracts of R. officinalis were 2.78, 15.87 and 15.70 ml/mg, respectively. On the other hand, antifeedant effect of methanol, ethylacetat and n-Hexan for R. officinalis estimated as 43.13%, 55.11% and 9.19%, respectively. All the obtained results revealed that methanol and ethylacetat extracts of R. officinalis are effective extracts for controlling the cotton bollworm population.

Keywords: Helocoverpa armigera, Rosemarinus officinalis, extract, methanol, ethylacetat, n-Hexan

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53 Characteristic Components in Cornusofficinalis to AGEs Injury Protective Effect and Mechanism of HUVEC

Authors: Yu-Han Tao, Hui-Qin Xu

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The present study aimed to explain the protective effect of Cornus officinalis characteristic components, under AGEs damage to HUVEC. After cultured HUVEC adhered, Cornus officinalis characteristic components such as loganin, morroniside, oleanolic acid, ursolic acid and aminoguanidine (positive control dug) hatched, after 1h the AGEs (200 mg/L) were added. After 24h, LDH, SOD, MDA, NO, ET, and AngⅡ, TGF-β, IL-1β, ROS in the supernatant were determined. The results showed the Cornus officinalis characteristic compounds could improve vitality of SOD, NO, reduce the MDA, ET, AngⅡ, TGF-β, IL-1β, ROS significantly when compared with the model groug. Loganin, oleanic acid, ursolic acid, had significant protective effect on AGEs injured HUVEC. As a conclusion, characteristic components in Cornus officinalis had a positive effect after HUVEC injured by AGEs.

Keywords: Cornus officinalis, morroniside, oganin, oleanolic acid, ursolic acid

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52 Literature Review on the Antibacterial Effects of Salvia officinalis L.

Authors: Benguerine Zohra, Merzak Siham, Pr. Chelghoum

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Introduction: The widespread production and consumption of antibiotics have raised significant concerns due to various adverse effects and the development of bacterial resistance. This increasing resistance to currently available antibiotics necessitates the search for new antibacterial agents. One alternative strategy to combat antibiotic-resistant bacteria is the use of natural antimicrobial substances such as plant extracts. This study aims to provide an overview of the antibacterial effects of Salvia officinalis (sage), a plant native to the Middle East and Mediterranean regions. Materials and Methods: This review was conducted by searching studies in databases such as PubMed, Scopus, JSTOR, and SpringerLink. The search terms were “Salvia officinalis L.” and “antibacterial effects.” Only studies that met our inclusion criteria (in English, focusing on the antibacterial effects of Salvia officinalis L., and primarily dated from 2012 to 2023) were considered for further review. Results and Discussion: The initial search strategy identified approximately 78 references, of which only 13 articles were included in this review. The synthesis of these articles revealed that multiple data sources confirm the antimicrobial effects of S. officinalis. Its essential oil and alcoholic extract exhibit strong bactericidal and bacteriostatic effects against both Gram-positive and Gram-negative bacteria. Conclusion: The significant value of the extract, oil, and leaves of S. officinalis demands further studies on other useful and unknown properties of this multipurpose plant.

Keywords: salvia officinalis, literature review, antibacterial., botany

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51 Literature Review of the Antibacterial Effects of Salvia Officinalis L.

Authors: Benguerine Zohra, Merzak Siham, Bouziane Cheimaa, Si Tayeb Fatima, Jou Siham, Belkessam

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Introduction: Antibiotics, widely produced and consumed in large quantities, have proven problematic due to various types of side effects. The development of bacterial resistance to currently available antibiotics has made the search for new antibacterial agents necessary. One alternative strategy to combat antibiotic-resistant bacteria is the use of natural antimicrobial substances such as plant extracts. The objective of this study is to provide an overview of the antibacterial effects of a plant native to the Middle East and Mediterranean regions, Salvia officinalis (sage). Materials and Methods: This review article was conducted by searching studies in the PubMed, Scopus, JSTOR, and SpringerLink databases. The search terms were "Salvia officinalis L." and "antibacterial effects." Only studies that met our inclusion criteria (in English, antibacterial effects of Salvia officinalis L., and primarily dating from 2012 to 2023) were accepted for further review. Results and Discussion: The initial search strategy identified approximately 78 references, with only 13 articles included in this review. The synthesis of the articles revealed that several data sources confirm the antimicrobial effects of S. officinalis. Its essential oil and alcoholic extract exhibit strong bactericidal and bacteriostatic effects against both Gram-positive and Gram-negative bacteria. Conclusion: The significant value of the extract, oil, and leaves of S. officinalis calls for further studies on the other useful and unknown properties of this multi-purpose plant.

Keywords: salvia officinalis, literature review, antibacterial, effects

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50 Effect of Dietary Melissa officinalis Leaves Supplementation on Lipid Oxidation of Broiler Breast Fillets During Refrigerated Storage

Authors: Khosro Ghazvinian, Touba Khodaeian

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To improve the oxidative stability of meat products, the use of dietary form of antioxidants can extend the shelf life and acceptability of muscle food during exposition or storage condition. As shown, this method is more effective than adding direct preservatives due to uniform incorporation of dietary additives into sub cellular membrane and therefore, they can properly inhibit the oxidative reaction at their localized sites. Furthermore, postmortem addition of antioxidants to meat cannot directly inhibit the oxidation in membrane phospholipids. Therefore, this study was designed to evaluate the effects of feed supplementation with Melissa officinalis leaves on lipid peroxidation of chicken breast fillets during refrigerated storage. In this study, 72 one-day old Ross 308 broilers distributed in four groups with six replicates (3 chickens each) were fed a basal diet (CONT) or basal diet supplemented with 5, 10, and 15 gr/Kg M.officinalis, for 6 weeks. Following slaughter, fillets from breast were stored at 4 °C in the dark for 12 days, and lipid oxidation was assessed on the basis of thiobarbituric acid reactive substances (TBARS) formed. Results showed that incorporation of M.officinalis in broiler diets delayed lipid oxidation in raw breast meat during refrigerated storage comparative with CONT(p<0.05). In this regard, TBARS levels of breast samples containing higher concentrations (10 and 15 gr/Kg) of M. officinalis (625.43 and 504.32 µg/kg MDA equivalents, respectively )were significantly lower than those of control and 5g/kg samples (872.75 and 841.32 µg/kg MDA equivalents, respectively) (p<0.05). Therefore, M. officinalis might be utilized in novel applications as a nutritional supplement or a functional food component.

Keywords: breast fillet, lipid oxidation, Melissa officinalis, TBARS assay

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49 Synergistic Interactions between Secondary Metabolites in Rosmarinus officinalis L.

Authors: Ruta Mickiene, Audrius Maruska, Ona Ragazinskiene

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This research focuses on phytochemistry and antimicrobial activities of compounds isolated and identified from species Rosmarinus officinalis L. This is a study of synergistic effects between phenolic fraction and essential oils. The antimicrobial activity of extracts from Rosmarinus officinalis L. originated from the sector of medicinal plants, Kaunas botanical garden of Vytautas Magnus University Lithuania, were tested by the method of series dilutions, against different bacteria species. Investigated microorganisms were Escherichia coli, Proteus vulgaris and Staphylococcus aureus with and without antibiotic resistances originating from livestock. The antimicrobial activities of extracts were described by determination of the Minimal Inhibitory Concentration (MIC). Preliminary results show that the MIC range between 9.0 % and 12.0 % for the different Rosmarinus officinalis L. extracts and bacterial species. The total amounts of phenolic compounds and total amounts of flavonoids were tested in the methanolic extracts of the plants. The chemical composition for essential oils analysed by GC/MS. Predominant components were alpha pinene (20%), camphor (10%), 1.8‐cineole (5%), phellandrene (6%), camphene (5%), beta pinene (4%), bornylacetate (4%), limonene (2%), borneol (3%), alpha terpineol (3%), cymene (2%), caryophyllene (15%), verbenone (7%), alpha terpinene (3%), eucalyptol (11%).

Keywords: antimicrobial activity, essential oil, Rosmarinus officinalis L., escherichia coli

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48 The Antimicrobial Activity of the Essential Oil of Salvia officinalis Harvested in Boumerdes

Authors: N. Mezıou-Cheboutı, A. Merabet, N. Behidj, F. Z. Bissaad

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The Algeria by its location, offers a rich and diverse vegetation. A large number of aromatic and medicinal plants grow spontaneously. The interest in these plants has continued to grow in recent years. Their particular properties due to the essential oil fraction can be utilized to treat microbial infections. To this end, and in the context of the valuation of the Algerian flora, we became interested in the species of the family Lamiaceae which is one of the most used as a global source of spices and extracts strong families antimicrobial potency. The plant on which we have based our choice is a species of sage "Salvia officinalis" from the Isser localized region within the province of Boumerdes. This work focuses on the study of the antimicrobial activity of essential oil extracted from the leaves of salvia officinalis. The extraction is carried out by HE hydrodistillation and reveals a yield of 1.06℅. The study of the antimicrobial activity of the essential oil by the method of at aromatogramme shown that Gram positive bacteria are most susceptible (Staphylococcus aureus and Bacillus subtilis) with a strong inhibition of growth. The yeast Candida albicans fungus Aspergillus niger and have shown moderately sensitive.

Keywords: Salvia officinalis, steam distillation, essential oil, aromatogram, anti-microbial activity

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47 Antimicrobial Activity of Some Alimentary and Medicinal Plants

Authors: Akrpoum Souad, Lalaoui Korrichi

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Vicia faba L.,Vaccinium macrocarpon, Punica granatum, Lavandula officinalis, Artemisia absinthium, Linum capitatum and Camellia sinensis were frequently used in our alimentation. In this study, we have tested the antimicrobial activity of their ethanolic and methanolic extracts on some pathogen bacteria, then their ability to in vivo inhibit the growth of Strepcoccus pneumonia. The phytochemical screening has given the composition of the most active extracts. According to the obtained results, the ethanolic extract of Lavendula. officinalis and A absinthium has shown an inhibition of all the tested strains of becteria3. The ethanolic extract of L. officinalis has given the highest activity against S. pneumoniae, followed by the methanolic extract of C. sinensis 1, 2 and P. granatum. The phytochemical screening showed that the most active extracts contained mainly naturels compounds.

Keywords: plants, extracts, antimicrobial activity, streptococcus pneumoniae, phytochemical screening

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46 Emblica officinalis Fruit Extract Ameliorates Cisplatin-Induced Nephrotoxicity in Experimental Rats

Authors: Prerna Kalra, Surender Singh

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Cisplatin is the most common chemotherapeutic agent used in different solid tumors, but its main limiting factor is dose-dependent nephrotoxicity by generating reactive oxygen species, by stimulating inflammatory and apoptotic pathways. Additional adjuvant therapies to decrease the toxicity of this chemotherapeutic drug are essential. This study was designed to evaluate the protective role of Emblica officinalis Geartn (Indian gooseberry) against cisplatin induced nephrotoxicity. Emblica officinalis was orally administered to Wistar rats (n=6) for 10 days in 50, 100 and 200mg/kg body weight. On day 7, 8mg/kg of cisplatin was administered intra-peritoneally to rats in all groups. Serum creatinine, blood urea nitrogen and antioxidant levels were measured on day10. The renal damage was evaluated by histopathological and transmission electron microscopy. We found that 200mg/kg dose of Emblica officinalis significantly inhibited the elevation of biochemical parameters i.e. serum creatinine, blood urea nitrogen, oxidant stress marker (malondialdehyde) and increased the reduced levels of antioxidant marker (endogenous glutathione and superoxide dismutase). Cisplatin treated rats have shown acute tubular necrosis and infiltration of inflammatory cells in rat kidney which was reversed after treating the animals with Emblica officinalis in the treatment group. In ultrastructural changes cisplatin treated group showed the damaged mitochondria (M) with dissolved cristae and large number of lysosomes (L) and vacuole (V) formation in tubular epithelial cells. EOE administered group showed visible cristae formation and sign of autophagy vacuoles at a dose of 200mg/kg. Further in-silico studies revealed that ellagic acid is responsible for its nephroprotective effect. The above findings conclude that the Emblica officinalis may be used as an adjuvant therapy in cisplatin induced nephrotoxicity.

Keywords: antioxidant, cisplatin, Emblica officinalis, in silico, nephrotoxicity

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45 Genotoxic and Cytotoxic Effects of Salvia officinals Extracts on Rat Bone Marrow

Authors: Mohammed A. Alshehri

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Salvia officinalis is an aromatic plant member of the mint (Labiatae) family. It is popular kitchen herb. Not surprise to find that the name of this herb related to cure, in Latin language Salvia means to cure where officinalis means medicinal which answer why the sage has a top place in the list of medicinal plants. The aim of the present study was to assess the genetic damage and cytological changes caused by exposure of the test organism (Rattusrattus) to Salvia officinals. For this purpose, adult female rats, weighing 200–250 g, were used as donors. A total of 36 adult Wister male rats were randomly assigned to five groups: the experimental groups (rats were intraperitonealy injected with Salvia officinalis pure extract at (0.1, 0.2, 0.5, 0.1mg/kg body weight, the same dose was administered once a day. Control group (rats were injected intraperitonealy physiological saline. And positive control were injected with Cyclophosphamide. On the 21st days following Salvia officinalis pure extract exposure, rats were sacrificed, and samples of bone marrow were collected. Following that, we performed a micronuclei (MN) test using MNNCE (Micro-nucleated normocromatic erythrocytes) and MNPCE (Micronucleated polychromatic erythrocytes), NDI (Nuclear division index), and cytological parameters using NDCI (nuclear division cytotoxicity index), necrotic, and apoptotic cells in rat's bone marrow samples. Results showed that there was a no significant increase in the frequency of micro-nucleatedas well as in cytological parameters in bone marrow cells. In light of these results, if Salvia officinalis pure extract may considered to be safe from the stand point of genotoxicity and cytotoxicity effects.

Keywords: Salvia officinalis, micronucleus, NDI, NDCI, toxicity, chromosomal aberrations

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44 Phytochemical Study and Biological Activity of Sage (Salvia officinalis L.)

Authors: Mekhaldi Abdelkader, Bouzned Ahcen, Djibaoui Rachid, Hamoum Hakim

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This study presents an attempt to evaluate the antioxidant and antimicrobial activity of methanolic extract and essential oils prepared from the leaves of sage (Salvia officinalis L.). The content of polyphenols in the methanolic extract of the leaves from Salvia officinalis extract was determined by spectrophoto- metrically, calculated as gallic acid and catechin equivalent. Antioxidant activity was evaluated by free radical scavenging activity using 2,2-diphenylpicryl-1-picrylhydrazyl (DPPH) assay. The plant essential oil and methanol extract were also subjected to screenings for the evaluation of their antioxidant activities using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) test. While the plant essential oil showed only weak antioxidant activities, its methanol extract was considerably active in DPPH (IC50= 37.29µg/ml) test. Appreciable total phenolic content (31.25mg/g) was also detected for the plant methanol extract as gallic acid equivalent in the Folin–Ciocalteu test. The plant was also screened for its antimicrobial activity and good to moderate inhibitions were recorded for its essential oil and methanol extract against most of the tested microorganisms. The present investigation revealed that this plant has rich source of antioxidant properties. It is for this reason that sage has found increasing application in food formulations.

Keywords: antibacterial activity, antioxidant activity, flavonoid, polyphenol, salvia officinalis

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43 Extraction, Isolation and Comparative Phtochemical Study of Aegle Marmelos, Calendula Officinalis and Fenugreek

Authors: Nitin Rajan, Kashif Shakeel, Shashank Tiwari, Shachan Sagar

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Background: - Aegle Marmelos (Bael) leaf extract is taken twice daily to treat ophthalmia, ulcers, and intestinal worms, among other ailments. Poultice made from bael leaf is used in the treatment of eye conditions. The leaf juice has a variety of therapeutic applications, with the most notable being the treatment of diabetes. Fenugreek is used to cure red spots around the eyes, as well as to soften the throat and chest and to give relief from coughing. The use of this plant in the form of infusion, powder, pomade, and decoction has been extremely popular in Iranian traditional medicine. The plant may be used to wash one's vaginal linings. This plant is used as an emollient in the lack of appetite, treatment of pellagra, and gastrointestinal problems, as well as a general tonic. Calendula officinalis leaves are used to treat varicose veins on the outside of the body by infusing them. In Europe, the leaves are diaphoretic and resolvent in nature, while the blooms are employed as an emmenagogue and antispasmodic stimulant in Canada and the United States. The flowers were decocted and served as a posset drink when smallpox and measles were common in England, and the fresh juice was used to treat jaundice. Objective: - This study is done to compare the physicochemical parameter of the alcoholic extract of the leaves of Aegle Marmelos, Calendula Officinalis, and Fenugreek. Materials and Methods: Extraction and Isolation of Aegle Marmelos, Calendula Officinalis, Fenugreek, were done. Preliminary phytochemical study for alkaloids, cardiac glycosides, flavonoids, glycosides, phenols, resins, saponins, steroids, tannins, terpenoids of the extract was done individual by using the standard procedure. Result: - The phytochemical screening of Aegle Marmelos, Calendula Officinalis, and Fenugreek shows the presence of alkaloids, carbohydrates, total phenolics, total flavonoids, tannins, saponins gum. Conclusion: - In this study, we have found that crude aqueous and organic solvent extracts of Aegle Marmelos, Calendula Officinalis, and Fenugreek leaves contain some important bioactive compounds and it justifies their use in the traditional medicines for the treatment of different diseases.

Keywords: Aegle Marmelos, Calendula Officinalis, Fenugreek, physiochemical parameter

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42 Impact of a Biopesticide Formulated an Entomopathogenic Fungus Metarhizium Anisopliae et Abstracts of Two Different Plants Sage (Salvia officinalis) and American Paper (Schinus molle) on Aphis Fabae (Homoptera - Aphididae)

Authors: Hicham Abidallah

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In this work we realized a formulation of an entomopathogenic fungus Metarhizium anisopliae with a dose of 1,7 x 105 spores/ml, and aqueous abstracts of two different plants sage (Salvia officinalis) and American paper (Schinus molle) with they’re full dose and half dose, on a black bean aphid populations (Aphis fabae) on a bean crop planted in pots at semi-controlled conditions. Five formulations were achieved (Met, Fd, F1/2d, Sd et S1/2d) and tested on six blocks each one contained six pots. This study revealed that four (04) formulations exercised an influence over black bean aphid (Met, Fd, F1/2d, Sd), of which Metarhizium marked the most elevated and aggressive toxicity with an efficiency of 99,24%, however, sage formulation with the half dose (S1/2d ) marked a weak toxicity with an efficiency of 18%. Test of Metarhizium anisopliae on bees didn’t show toxicity, and no mortality has been marked, and no trace of green Muscardine observed.

Keywords: Metarhizium anisopliae, salvia officinalis, Schinus molle, Aphis fabae, efficiency degree

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41 Essential Oil Composition and Antimicrobial Activity of Rosmarinus officinalis L. Grown in Algeria (Djelfa)

Authors: Samah Lakehal, A. Meliani, F. Z. Benrebiha, C. Chaouia

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In the last few years, due to the misuse of antibiotics and an increasing incidence of immunodeficiency-related diseases, the development of microbial drug resistance has become more and more of a pressing problem. Recently, natural products from medicinal plants represent a fertile ground for the development of novel antibacterial agents. Plants essential oils have come more into the focus of phytomedicine. The present study describes antimicrobial activity of Rosmarinus officinalis L. essential oil known medicinally for its powerful antibacterial properties. The essential oil of rosemary obtained by hydrodistillation (using Clevenger type apparatus) growing in Algeria (Djelfa city of south Algeria) was investigated by GC-MS. The essential oil yield of the study was 1.4 %. The major components were found to be camphor, camphene, 1,8-cineole. The essential oil has been tested for antimicrobial activity against eight bacteria (Gram-negative and Gram-positive), and three fungi including Candida albicans. Inhibition of growth was tested by the agar diffusion method based on the determination of the diameter of inhibition. The oil was found to have significant antibacterial activity and therefore can be used as a natural antimicrobial agent for the treatment of several infectious diseases caused by those germs, which have developed resistance to antibiotics.

Keywords: antimicrobial activity, Rosmarinus officinalis L., essential oils, GC/MS, camphor

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40 In vitro Effects of Salvia officinalis on Bovine Spermatozoa

Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

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In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa

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39 Evidence of Microplastics Ingestion in Two Commercial Cephalopod Species: Octopus Vulgaris and Sepia Officinalis

Authors: Federica Laface, Cristina Pedà, Francesco Longo, Francesca de Domenico, Riccardo Minichino, Pierpaolo Consoli, Pietro Battaglia, Silvestro Greco, Teresa Romeo

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Plastics pollution represents one of the most important threats to marine biodiversity. In the last decades, different species are investigated to evaluate the extent of the plastic ingestion phenomenon. Even if the cephalopods play an important role in the food chain, they are still poorly studied. The aim of this research was to investigate the plastic ingestion in two commercial cephalopod species from the southern Tyrrhenian Sea: the common octopus, Octopus vulgaris (n=6; mean mantle length ML 10.7 ± 1.8) and the common cuttlefish, Sepia officinalis (n=13; mean ML 13.2 ± 1.7). Plastics were extracted from the filters obtained by the chemical digestion of cephalopods gastrointestinal tracts (GITs), using 10% potassium hydroxide (KOH) solution in a 1:5 (w/v) ratio. Once isolated, particles were photographed, measured, and their size class, shape and color were recorded. A total of 81 items was isolated from 16 of the 19 examined GITs, representing a total occurrence (%O) of 84.2% with a mean value of 4.3 ± 8.6 particles per individual. In particular, 62 plastics were found in 6 specimens of O. vulgaris (%O=100) and 19 particles in 10 S. officinalis (%O=94.7). In both species, the microplastics size class was the most abundant (93.8%). Plastic items found in O. vulgaris were mainly fibers (61%) while fragments were the most frequent in S. officinalis (53%). Transparent was the most common color in both species. The analysis will be completed by Fourier transform infrared (FT-IR) spectroscopy technique in order to identify polymers nature. This study reports preliminary data on plastic ingestion events in two cephalopods species and represents the first record of plastic ingestion by the common octopus. Microplastic items detected in both common octopus and common cuttlefish could derive from secondary and/or accidental ingestion events, probably due to their behavior, feeding habits and anatomical features. Further studies will be required to assess the effect of marine litter pollution in these ecologically and commercially important species.

Keywords: cephalopods, GIT analysis, marine pollution, Mediterranean sea, microplastics

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38 Antibacterial Effect of Hydroalcoholic Extract of Salvia Officinalis and, Mentha Pulegium on Three Strains of Streptococcus Mutants, Lactobacillus Rhamnosus and, Actinomyces Viscosus Dental Caries in-vitro

Authors: H. Nabahat, E. Amiri, F. AzaditalabDavoudabadi, N. Zaeri

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Tooth decay is one of the most common forms of oral and dental illness in the world, which causes huge costs of treatment, especially in high-risk groups such as people with oral dry mouth, prevention and control of it are very important. The use of traditional treatments such as extraction of drugs from medicinal plants is of paramount importance to Iran and the international community as well. The present study was conducted with the aim of investigating the antibacterial effect of the extract of Salvia officinalis and Mentha pulegium, which are the most commonly used drugs in the treatment of oral and teeth bacterial (Streptococcus mutant, Lactobacillus rhamnosis, and Actinomyces viscosis) in vitro method. In this experimental study, two herbs of Salvia and Mentha were prepared by maceration of hydroalcoholic extract, and the antibacterial effect was evaluated by broth macro dilution on streptococcal mutagen bacteria, lactobacillus rhamnosis, and viscose actinomycosis. The results were analyzed by the Whitney Mann test (P > 0.05). The results showed that the minimum inhibitory concentration (MIC) of the salmonella extract for Streptococcus mutan were 6.25 and 12.5 μg/ml, respectively, for lactobacillus of 1.56 and 3.12 μg/ml, respectively, and for actinomycosis viscose, The order of 12.5 and 100 μg/ml was obtained. As a result, broth macro dilution showed that both extracts of Salvia and Mentha had an inhibitory effect on all three species of bacteria. This effect for Salvia was significantly (P < 0.05) more than Mentha and was within the concentration range of both the extracts and had a bactericidal effect on all three bacteria.

Keywords: antibacterial effect, dental bacteria, herbal extracts , salvia officinalis, mentha pulegium

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37 Egg Hatching Inhibition Activity of Volatile Oils Extracted from Some Medicinal and Aromatic Plants against Root-Knot Nematode Meloidogyne hapla

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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36 Antimicrobial Potential of Calendula officinalis Extracts on Flavobacterium columnare of Clarias gariepinus Fingerlings

Authors: Nelson Rotimi Osungbemiro, Sanni Rafiu Olugbenga, Abayomi Olufemi Olajuyigbe

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Ninety Fingerlings of Clarias gariepinus were exposed to the pathogenic Flavobacterium columnare a Gram Negative bacteria responsible for high mortality in fish pond raised young fish (fries and fingerlings) of Clarias sp. in Southwestern Nigeria. After feeding with 40% crude protein pelletized fish feed for 5 days, the fishes were divided into two groups, one group was treated with extracts from Calendula officinalis flowers, while the second group was not treated (control). The results indicated that, at day 5, colony formation had been manifesting and at day 7, skin lesion occurred and at the 8th day, first mortality of fish occurred, and this continued steadily on the 9th-12th day when all the fishes were dead. Whereas, in the group that was treated with Calendula sp., no single mortality was recorded. This research shows that plant extract from Calendula flowers is an effective antimicrobial agent against the virulent pathogenic Flavobacterium columnare disease.

Keywords: antimicrobial, Flavobacterium columnare, Clarias gariepinus, fish

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35 Antioxidant Activity of the Methanolic Extract and Antimicrobial Activity of the Essential Oil of Rosmarinus officinalis L. Grown in Algeria

Authors: Nassim Belkacem, Amina Azzam, Dalila Haouchine, Kahina Bennacer, Samira Soufit

Abstract:

Objective: To evaluate the antioxidant activity of the methanolic extract along with the antimicrobial activity of the essential oil of the aerial parts of Rosmarinus officinalis L. collected in the region of Bejaia (northern center of Algeria). Materials and methods: The polyphenols and flavonoids contents of the methanolic extract were measured. The antioxidant activity was evaluated using two methods: the ABTS method and DPPH assay. The antimicrobial activity was studied by the agar diffusion method against five bacterial strains (Three Gram positive strains and two Gram negative strains) and one fungus. Results: The total polyphenol and flavonoid content was about 43.8 mg gallic acid equivalent per gram (GA Eq/g) and 7.04 mg quercetin equivalent per gram (Q Eq/g), respectively. In the ABTS assay, the rosemary extract has shown an inhibition of 98.02% at the concentration of 500ug/ml with a half maximal inhibitory concentration value (IC50) of 194.92ug/ml. The results of DPPH assay have shown that the rosemary extract has an inhibition of 94.67 % with an IC50 value of 17.87ug/ml, which is lower than that of Butylhydroxyanisol (BHA) about 6.03ug/ml and ascorbic acid about 1.24μg/ml. The yield in essential oil of rosemary obtained by hydrodistillation was 1.42%. Based on the determination of the diameter of inhibition, different antimicrobial activity of the essential oil was revealed against the six tested microbes. Escherichia coli from the University Hospital (UH), Streptococcus aureus (UH) and Pseudomonas aeruginosa ATCC have a minimum inhibitory concentration value (MIC) of 62.5µl/ml. However, Bacillus sp (UH) and Staphylococcus aureus ATCC have an MIC value of 125μl/ml. The inhibition zone against Candida sp was about 24 mm. The aromatograms showed that the essential oil of rosemary exercises an antifungal activity more important than the antibacterial one.

Keywords: Rosmarinus officinalis L., maceration, essential oil, antioxidant, antimicrobial activity

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34 Evaluation of Calendula officinalis L. Flower Dry Weight, Flower Diameter, and Number of Flower in Plant Variabilities under Effect of Compost and Nitrogen Different Levels in Four Harvest

Authors: Amin Rezazadeh, Parisa Farahpour, Arezoo Rezazadeh, Morteza Sam Deliri

Abstract:

In order to investigate the effects of nitrogen and compost different levels on qualitative and quantitative performance of Calendula officinalis L. herb, an experiment was carried out in the research field of Chalous Azad University in 2011-2012. The experiment was done in factorial form as a randomized complete block design, in three replicates. Treatments consisted of nitrogen and compost. Considered nitrogen levels consisted of N0=0, N1=50, N2=100 kg/ha and compost levels were including C0=0, C1=6, C2=12 ton/ha. Investigated characteristics consisted of flower dry weight, number of flowers in plant, flower diameter. The results showed, nitrogen and compost treatments had statistically significant influence (p ≤ 0.01) on studied characteristics. Flower dry weight, flower diameter and number of flower in plant characteristics has been studied in four harvest; as, the performance of these characteristics had increasing procedure from the first harvest up to the forth harvest; and, in the fourth harvest, it has reached to its` maximum level. As, up to the forth harvest, the maximum flower dry weight, flower diameter and number of flower in plant obtained by C1× N2 (C1=6 ton/ha compost and N2=100 kg/ha nitrogen) treatment.

Keywords: calendula, compost, nitrogen, flavonoid

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33 Health Promoting Properties of Phytochemicals from Rosemary (Rosmarinus officinalis) for Cancer and Inflammatory Bowel Disease

Authors: Jeremy J. Johnson

Abstract:

Mediterranean herbs including rosemary (Rosmarinus officinalis) contain a variety of phytochemicals including diterpenes that possess extensive biological activity. Applications of diterpenes, including the more abundant forms carnosol and carnosic acid, have been shown to possess anti-cancer, anti-inflammatory, anti-oxidant, and anti-proliferation properties. To confirm these properties, we have evaluated rosemary extract and selected diterpenes for biological activity in cancer and inflammatory models. Our preliminary data have revealed that select diterpenes can disrupt androgen receptor functionality in prostate and breast cancer cells. This property is unique among natural products for hormone-responsive cancers. The second area of interest has been evaluating rosemary extract and selected diterpenes for activation of sestrin-2, an antioxidant protein, in colon cancer cells. A combination of in vitro and in vivo approaches have been utilized to characterize the activity of rosemary diterpenes in rosemary. Taken together, these results suggest that phytochemicals found in rosemary have distinct pharmacological actions for disrupting cell-signaling pathways in cancer and inflammatory bowel disease.

Keywords: rosemary, diterpene, cancer, inflammation

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32 Study of Effects of Hydro-Alcoholic Extract of Asparagus Root (Asparagus officinalis) Ontestes Spermyogenesis Index of Laboratory Mouse

Authors: Hamid Karimi, Naegar Mahdavi, Hossein Tayefi Nasrabadi

Abstract:

Spermatozoids production rate and its quality are more important factors in the diagnosis of infertility. Also, spematozids activity have a more important role in fertilization. Some medicinal plants as Asparagus(Asparagus officinalis) has many antioxidant component. Therefore, They can affect testes tissue to production more and high-quality spermatozoids. In this survey, Asparagus root extract is studied on spermatogenesis index in the laboratory mouse testes. Hydro-alcoholic extract of asparagus root is prepared and examined on four group of the mature male mouse. Blank group without extract, group 1,100ml/kg dose, group 2, 200 ml/kg dose and group 3, 300ml/kg dose. Then, mice are euthanized, and testes are removed. Testes are weighted, and paraffinized blocks are prepared. TDI(Tubular Differentiation Index) and SPI(Spermiation Index) are studied on histological sections by light microscope. This study results were showed that TDI and SPI in treatments groups with 200 and 300 ml/kg dose had significant enhancement (P<0.05). Consequently, Extract of Asparagus root can enhance spermatozoid production and, therefore, cause improve fertility in male laboratory mice.

Keywords: histology, spermatozoid, ASP [aragus, testes

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31 Morroniside Intervention Mechanism of Renal Lesions, a Combination Model of AGEs Exacerbation of STZ-Induced Diabetes Mellitus

Authors: Hui-Qin Xu, Xing Lv, Yu-Han Tao

Abstract:

The depth study aimed on the mechanism of morroniside in protecting diabetic nephropathy. The diabetic mice models with blood glucose above 15mmol/L were divided into model, aminoguanidine, metformin, captopril, morroniside low-dose, and morroniside high-dose groups. And normal group was set simultaneously. All groups were fed with high AGEs food except normal group. Each group was intragastric administration of the corresponding medicine except model and normal groups. After 12 weeks, all the indictors were measured. It showed that the morroniside could reduce blood glucose significantly, urinary protein, serum urea nitrogen, creatine, pathological changes, AGEs levels, renal cortex RAGE mRNA and RAGE protein expression levels; increase food consumption, water intake, urine volume, insulin secretion. As a conclusion, morroniside from cornus officinalis can protect renal in diabetic mice, its mechanism may be related to the proliferation of islet cells, rectify glycometabolism, reduce serum and kidney AGEs content, and descend renal RAGEmRNA and RAGE protein expression levels.

Keywords: cornus officinalis, diabetic nephropathy, morroniside, RAGE protein

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30 Effect of Organic Matter Pre-Treatment on Germination and Seedlings Growth Indices on Marigold (Calendula officinalis L.) under Drought Stress

Authors: Fahimeh Helali Soltanahmadi, Alireza Nourabdi

Abstract:

To investigate the effects of seed pretreatment on drought tolerance, an experimental Marigold (Calendula officinalis L) was tested in 2017-2018. A germination experiment was performed in the laboratory of Urmia University. The experiment was performed as a factorial experiment in a completely randomized design with three replications. The experimental treatments were dry levels of polyethylene glycol in four levels (control, -3, -6 and -9 bar) and treatments of Humic acid (72 mg / l), salicylic acid (2000 μmol) and ascorbic acid (200 ppm), and control were performed. The results of the germination experiment showed that with increasing drought levels caused by polyethylene glycol, germination indices were significantly reduced, so that the highest value obtained from the measured traits was observed in normal irrigation and in priming treatments, and the lowest amount of these traits was observed at the -9 bar drought level due to polyethylene glycol in the control treatment. The results of this study showed an increase in drought tolerance and improved Marigold germination under stress by applying seed priming with salicylic acid, which is recommended in sustainable and organic agriculture.

Keywords: Marigold, organic matter, seed pretreatment, drought stress, allometric coefficient

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29 Examinations of Sustainable Protection Possibilities against Granary Weevil (Sitophilus granarius L.) on Stored Products

Authors: F. Pal-Fam, R. Hoffmann, S. Keszthelyi

Abstract:

Granary weevil, Sitophilus granarius (L.) (Col.: Curculionidae) is a typical cosmopolitan pest. It can cause significant damage to stored grains, and can drastically decrease yields. Damaged grain has reduced nutritional and market value, weaker germination, and reduced weight. The commonly used protectants against stored-product pests in Europe are residual insecticides, applied directly to the product. Unfortunately, these pesticides can be toxic to mammals, the residues can accumulate in the treated products, and many pest species could become resistant to the protectants. During recent years, alternative solutions of grain protection have received increased attention. These solutions are considered as the most promising alternatives to residual insecticides. The aims of our comparative study were to obtain information about the efficacies of the 1. diatomaceous earth, 2. sterile insect technology and 3. herbal oils against the S. granarius on grain (foremost maize), and to evaluate the influence of the dose rate on weevil mortality and progeny. The main results of our laboratory experiments are the followings: 1. Diatomaceous earth was especially efficacious against S. granarius, but its insecticidal properties depend on exposure time and applied dose. The efficacy on barley was better than on maize. Mortality value of the highest dose was 85% on the 21st day in the case of barley. It can be ascertained that complete elimination of progeny was evidenced on both gain types. To summarize, a satisfactory efficacy level was obtained only on barley at a rate of 4g/kg. Alteration of efficacy between grain types can be explained with differences in grain surface. 2. The mortality consequences of Roentgen irradiation on the S. granarius was highly influenced by the exposure time, and the dose applied. At doses of 50 and 70Gy, the efficacy accepted in plant protection (mortality: 95%) was recorded only on the 21st day. During the application of 100 and 200Gy doses, high mortality values (83.5% and 97.5%) were observed on the 14th day. Our results confirmed the complete sterilizing effect of the doses of 70Gy and above. The autocide effect of 50 and 70Gy doses were demonstrated when irradiated specimens were mixed into groups of fertile specimens. Consequently, these doses might be successfully applied to put sterile insect technique (SIT) into practice. 3. The results revealed that both studied essential oils (Callendula officinalis, Hippophae rhamnoides) exerted strong toxic effect on S. granarius, but C. officinalis triggered higher mortality. The efficacy (94.62 ± 2.63%) was reached after a 48 hours exposure to H. rhamnoides oil at 2ml/kg while the application of 2ml/kg of C. officinalis oil for 24 hours produced 98.94 ± 1.00% mortality rate. Mortality was 100% at 5 ml/kg of H. rhamnoides after 24 hours duration of its application, while with C. officinalis the same value could be reached after a 12 hour-exposure to the oil. Both essential oils applied were eliminated the progeny.

Keywords: Sitophilus granarius, stored product, protection, alternative solutions

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28 Feasibility Studies on the Removal of Fluoride from Aqueous Solution by Adsorption Using Agro-Based Waste Materials

Authors: G. Anusha, J. Raja Murugadoss

Abstract:

In recent years, the problem of water contaminant is drastically increasing due to the disposal of industrial wastewater containing iron, fluoride, mercury, lead, cadmium, phosphorus, silver etc. into water bodies. The non-biodegradable heavy metals could accumulate in the human system through food chain and cause various dreadful diseases and permanent disabilities and in worst cases it leads to casual losses. Further, the presence of the excess quantity of such heavy metals viz. Lead, Cadmium, Chromium, Nickel, Zinc, Copper, Iron etc. seriously affect the natural quality of potable water and necessitates the treatment process for removal. Though there are dozens of standard procedures available for the removal of heavy metals, their cost keeps the industrialists away from adopting such technologies. In the present work, an attempt has been made to remove such contaminants particularly fluoride and to study the efficiency of the removal of fluoride by adsorption using a new agro-based materials namely Limonia acidissima and Emblica officinalis which is commonly referred as wood apple and gooseberry respectively. Accordingly a set of experiments has been conducted using batch and column processes, with the help of activated carbon prepared from the shell of wood apple and seeds of gooseberries. Experiments reveal that the adsorption capacity of the shell of wood apple is significant to yield promising solutions.

Keywords: adsorption, fluoride, agro-based waste materials, Limonia acidissima, Emblica officinalis

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27 Evaluation of Chemical Compositions and Biological Activities of Five Essential Oils

Authors: G. Ozturk, B. Demirci

Abstract:

It is well known that essential oils used for therapeutic purposes for many years. In this study, five different Pharmacopoeia grade essential oils (Achillea millefolium L., Pimpinella anisum L., Matricaria recutita L., Eucalyptus globulus L., Salvia officinalis L.) which obtained from commercial sources were evaluated for chemical compositions, synergistic antimicrobial activities, and lipoxygenase enzyme inhibitions. Volatile components were determined by gas chromatography/flame ionization detector and gas chromatography/mass spectrometer, simultaneously. The potential antimicrobial activity of essential oils was tested against oral pathogenic standard strains such as Streptococcus mutans, Streptococcus sanguinis, Staphylococcus aureus, Corynebacterium striatum, Candida albicans and Candida krusei by broth microdilution methods. Ciprofloxacin and ketoconazole were used positive controls. It has been observed that the essential oils tested have average inhibitory antimicrobial activity against oral pathogens with a Minimum Inhibition Concentration of 20-0.625 mg/mL. The active essential oils have been combined with antibiotics and synergistic effects have been evaluated by Checkerboard method. ƩFIC values were determined. In combination with antibiotics M. recutita essential oil has been shown to have a synergistic effect against S. aureus in combination with tetracycline (ƩFIC 0.46). In addition, 5-LOX inhibitory activity was measured by modifying the spectrophotometric method developed by Baylac and Racine. As a result, 5-LOX % inhibition of S. officinalis, E. globulus and M. recutita were calculated as 34.0 ± 6.66, 72.7 ± 2.78 and 27.7 ± 0.60, respectively.

Keywords: antimicrobial activity, essential oils, synergistic activity, 5-lipoxygenase inhibition

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26 An Efficient and Low Cost Protocol for Rapid and Mass in vitro Propagation of Hyssopus officinalis L.

Authors: Ira V. Stancheva, Ely G. Zayova, Maria P. Geneva, Marieta G. Hristozkova, Lyudmila I. Dimitrova, Maria I. Petrova

Abstract:

The study describes a highly efficient and low-cost protocol for rapid and mass in vitro propagation of medicinal and aromatic plant species (Hyssopus officinalis L., Lamiaceae). Hyssop is an important aromatic herb used for its medicinal values because of its antioxidant, anti-inflammatory and antimicrobial properties. The protocol for large-scale multiplication of this aromatic plant was developed using young stem tips explants. The explants were sterilized with 0.04% mercuric chloride (HgCl₂) solution for 20 minutes and washing three times with sterile distilled water in 15 minutes. The cultural media was full and half strength Murashige and Skoog medium containing indole-3-butyric acid. Full and ½ Murashige and Skoog media without auxin were used as controls. For each variant 20 glass tubes with two plants were used. In each tube two tip and nodal explants were inoculated. Maximum shoot and root number were obtained on ½ Murashige and Skoog medium supplemented with 0.1 mg L-1 indole-3-butyric acid at the same time after four weeks of culture. The number of shoots per explant and shoot height were considered. The data on rooting percentage, the number of roots per plant and root length were collected after the same cultural period. The highest percentage of survival 85% for this medicinal plant was recorded in mixture of soil, sand and perlite (2:1:1 v/v/v). This mixture was most suitable for acclimatization of all propagated plants. Ex vitro acclimatization was carried out at 24±1 °C and 70% relative humidity under 16 h illuminations (50 μmol m⁻²s⁻¹). After adaptation period, the all plants were transferred to the field. The plants flowered within three months after transplantation. Phenotypic variations in the acclimatized plants were not observed. An average of 90% of the acclimatized plants survived after transferring into the field. All the in vitro propagated plants displayed normal development under the field conditions. Developed in vitro techniques could provide a promising alternative tool for large-scale propagation that increases the number of homologous plants for field cultivation. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.

Keywords: Hyssopus officinalis L., in vitro culture, micro propagation, acclimatization

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25 Arbuscular Mycorrhizal Symbiosis Modulates Antioxidant Capacity of in vitro Propagated Hyssop, Hyssopus officinalis L.

Authors: Maria P. Geneva, Ira V. Stancheva, Marieta G. Hristozkova, Roumiana D. Vasilevska-Ivanova, Mariana T. Sichanova, Janet R. Mincheva

Abstract:

Hyssopus officinalis L., Lamiaceae, commonly called hyssop, is an aromatic, semi-evergreen, woody-based, shrubby perennial plant. Hyssop is a good expectorant and antiviral herb commonly used to treat respiratory conditions such as influenza, sinus infections, colds, and bronchitis. Most of its medicinal properties are attributed to the essential oil of hyssop. The study was conducted to evaluate the influence of inoculation with arbuscular mycorrhizal fungi of in vitro propagated hyssop plants on the: activities of antioxidant enzymes superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase; accumulation of non-enzymatic antioxidants total phenols and flavonoid, water-soluble soluble antioxidant metabolites expressed as ascorbic acid; the antioxidant potential of hyssop methanol extracts assessed by two common methods: free radical scavenging activity using free stable radical (2,2-diphenyl-1-picrylhydrazyl, DPPH• and ferric reducing antioxidant power FRAP in flowers and leaves. The successfully adapted to field conditions in vitro plants (survival rate 95%) were inoculated with arbuscular mycorrhizal fungi (Claroideoglomus claroideum, ref. EEZ 54). It was established that the activities of enzymes with antioxidant capacity (superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase) were significantly higher in leaves than in flowers in both control and mycorrhized plants. In flowers and leaves of inoculated plants, the antioxidant enzymes activity were lower than in non-inoculated plants, only in SOD activity, there was no difference. The content of low molecular metabolites with antioxidant capacity as total phenols, total flavonoids, and water soluble antioxidants was higher in inoculated plants. There were no significant differences between control and inoculated plants both for FRAP and DPPH antioxidant activity. According to plant essential oil content, there was no difference between non-inoculated and inoculated plants. Based on our results we could suggest that antioxidant capacity of in vitro propagated hyssop plant under conditions of cultivation is determined by the phenolic compounds-total phenols and flavonoids as well as by the levels of water-soluble metabolites with antioxidant potential. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.

Keywords: antioxidant enzymes, antioxidant metabolites, arbuscular mycorrhizal fungi, Hyssopus officinalis L.

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