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  3. Investigating the antifungal effect of the total methanolic extract and etherdopterylic, chloroformic, ethyl acetate, methanolic and aqueous fractions of the flowering aerial parts of Otostegiapersica (Burm.) Boiss

Investigating the antifungal effect of the total methanolic extract and etherdopterylic, chloroformic, ethyl acetate, methanolic and aqueous fractions of the flowering aerial parts of Otostegiapersica (Burm.) Boiss

Number of pages: 95 File Format: word File Code: 31986
Year: 2012 University Degree: Master's degree Category: Medical Sciences
Tags/Keywords: Antifungal effect - Chloroform - Etherdopterous fractions - Ethyl acetate - Golder - Mushroom suspension - Otostegia persica - Suspension - Total methanolic extract
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  • Summary of Investigating the antifungal effect of the total methanolic extract and etherdopterylic, chloroformic, ethyl acetate, methanolic and aqueous fractions of the flowering aerial parts of Otostegiapersica (Burm.) Boiss

    Dissertation

    To receive a doctorate in pharmacy

    Persian summary

    Otostegia persica is one of the plants of the Lamiaceae family (mint), which in traditional medicine is decoction of its aerial part to treat malaria, reduce blood sugar and blood pressure, and relieve fever

    It is used.

    The name of this plant in Persian culture is "Goldar". This genus has 3 species of perennial herbaceous plants in Iran:

    Otostegia aucheri, Otostegia micchauxii, Otostegia persica, among these 3 species, O.persica and O.micchauxi are endemic to Iran.

    The main compounds that have been studied in this plant are diterpenoids. Interesting therapeutic effects of this plant have been reported, such as antioxidant effect, antiglycation effect, blood sugar reduction, effect on morphine deprivation syndrome, significant antibacterial effect and the ability to inhibit acetylcholinesterase enzyme. We decided to investigate the above-mentioned effect for the first time on the total methanolic extract and ethereal, chloroform, ethyl acetate, methanolic and aqueous fractions of the flowering aerial part of the plant on 9 fungal strains: Trichophyton verrocosum, Epidermophyton floccosum, Microsporum gypseum Aspergillus flavus, Aspergillus fumigates, Candida albicans, Trichophyton mentagropheytes, Trichophyton rubrum, Microsporum cani.

    Methods of implementation:

    Aerial parts of Otostegia persica plant were collected on June 20, 1390 near Bandar Abbas. After collecting and scientific identification, the plant was powdered and its total methanolic extract was prepared by percolation method. Its ethereal, chloroform, ethyl acetate, methanolic and aqueous fractions were prepared by percolation method from the powdered plant. The obtained extract and fractions were concentrated and finally dried and kept in clean, dark containers in a cool place to check the antifungal effect. To investigate the effect of plant extracts and fractions on mushrooms, the well method has been used.

    In this way, extracts and desired fractions were poured into wells with a diameter of half a centimeter, and then the mushrooms were cultivated. To do this, the extracts obtained from the above plant were prepared by soaking method and after evaporation of the solvent, the extracts were prepared with concentrations of 200, 100, 50, 25, 12.5, 6.25 mg/ml with the help of water and ethanol 96 solvent. Then, the antifungal effect of the extracts was investigated by well method in Sabour dextrose agar and Sabour medium containing chloramphenicol and cyclohexamide. Antifungal drug was used for + control and solvent for - control.

    Checking other methods:

    Mixing method:

    To perform this method after preparing the culture medium, concentrations of 100mg/ml, 200mg, 50mg/ml, 25mg/ml 12.5mg/ml, 6.25mg/ml dissolved in sterile distilled water, prepared and poured into a warm culture medium and stirred, and the prepared suspension of the culture medium and extract was poured into the plates and left to cool, and after 24 hours, the desired mushrooms were cultured on the prepared culture medium.

    Mushroom suspension and plant extract method:

    To perform this method, the concentrations mentioned above were dissolved in 1 cc of distilled water and a specific amount of the desired mushrooms was poured into the prepared extracts and the suspension was left for 1 hour, and then this suspension was poured on the previously prepared and closed culture medium and the results were checked 7 days later.

    Results: Based on the investigations carried out in these The method of antifungal effect of this plant was not observed.

    Keywords: Otostegia persica, well, antifungal effect, suspension

    . Necessity and importance of the subject

    Plants have often been a unique source for drugs used by human society, and these natural drugs play an important role in the treatment of diseases.Necessity and importance of the topic

    Plants have often been a unique source for drugs consumed by human society, and these natural drugs play an important role in the treatment of diseases. One of the important problems that mankind has been dealing with since the beginning of creation is fungal diseases, and due to the undesirable side effects of synthetic drugs, the need to obtain new natural compounds with antifungal activity is felt.

    The plant Otostegia persica (Burm.) Boiss belongs to the mint family, considering that no mycological study has been done on it so far, we decided to determine the antifungal effect of the plant for the first time with standard methods and its antifungal effects on 9 fungal strains

    Microsporum gypseum, Epidermophyton foccosum, Trichophyton vorrocosum, Trichophyton mentagropheytes, Aspergillus fumigates, Aspergillus flavus, Microsporum canis, Trichophyton rubrum, Candida albicans.

    All parts of the country have diverse vegetation. The properties of many plants of this vegetation have been studied, and while many of them have not yet been researched and their effects are not known.

    On the other hand, synthetic drugs are not able to relieve or treat all problems. Today, many herbal and natural medicines in the fields of herbal therapy, homeopathy, aroma therapy, flower therapy and other complementary medicine methods are widely used and are used by millions of people in the world. Also, synthetic drugs have a specific and mostly one-dimensional effect due to the limitation of the effective substance, while herbal medicines contain a large number of substances that have a multidimensional effect and while creating different and complementary effects, they have few side effects. Because a large number of substances in plants are responsible for reducing or eliminating side effects.

    1- In 2009, 4 diterpenoids were extracted and their structure determined during a phytochemical study conducted on the aerial part of Otostegia persica.

    2- In 2005, a phytochemical research was conducted on O. persica for the first time. 3 Flavonols Kaempferol, morin and quercetin were identified in this plant. Kaempferol and quercetin aglycones are flavonols and are widely distributed in plants. (3)

    3- During a study conducted in 2010, the anti-glycation effect of this plant was proven, and a flavonoid named opatorin was extracted and its structure was determined in this regard (4).

    4- In 2010, the antidiabetic effect of this plant in Diabetic mice induced with streptozocin were proven (5).

    5- In 2004, the significant effect of this plant on morphine withdrawal syndrome was proven (6).

    6- In 2006, the studies conducted showed a significant antibacterial effect (7).

     

     

    1-3. The main objective

    Investigation of the antifungal effect of methanolic total extract and etherdopterol, chloroform, ethyl fractions

    acetate, methanol and water of floral aerial part Otostegiapersica (Burm.) Boiss

    Abstract

    Persica plant is in a Lamiaceae (mint) family. Since there is no biological work on antifungal activity of Otostegia persica, we prompted to study the antifungal activity of total methanolic extract and different fractions of the flowering aerial parts of this plant against 9 fungal strains including Trichophyton verrocosum, Epidermophyton floccosum, Microsporum gypseum, Aspergillus flavus, Aspergillus fumigates, Candida albicans, Trichophyton mentagropheytes, Trichophyton rubrum, Microsporum canis.

    Materials and methods: Flowering aerial parts of the plant were collected from Bandar Abbas.

  • Contents & References of Investigating the antifungal effect of the total methanolic extract and etherdopterylic, chloroformic, ethyl acetate, methanolic and aqueous fractions of the flowering aerial parts of Otostegiapersica (Burm.) Boiss

    List:

    Persian summary.. 1

    Chapter One: Generalities

     

    1-1. Necessity and importance of the topic.. 4

    1-2. Statement of the problem.. 4

    1-3. Main goal.. 5

    Chapter Two: Review Texts and studies of others in this field

    Part 1: Botany generalities

    2-1-1. Features of dark mints.. 8

    2-1-2. Classification of Otostegia persica plant. 10

    2-1-3. Otostegia genus.. 11

    2-1-4. Persica species.. 11

    2-1-5. Persica species habitat.. 12

    2-1-6. Pharmacological and biological effect of Otostegia genus. 12

    2-1-7. Uses of Otostegia genus. 13

     

     

    Part Two: Preparation steps of herbal extract

    2-2-1. Definition of extract.. 16

    2-2-2. Extraction.. 16

    2-2-3. Extraction methods... 16

    2-2-3-1. Soaking method.. 16

    2-2-3-2. Percolation method.. 17

    2-2-3-3. Selection of suitable solvent.. 18

    2-2-3-4. Advantages of percolation.. 19

    2-2-3-5. Soaking method with gentle heat. 19

    2-2-3-6. Brewing method .. 19

    2-2-3-7. Boiling method .. 20

    2-2-4. Different methods of extract concentration. 20

    2-2-4-1. Distillation in a vacuum.. 20

    2-2-4-2. Lyophilization.. 20

    2-2-4-3. Direct heat.. 21

     

    The third part: General mycology

    2-3-1. Characteristics of mushrooms.. 23

    2-3-2. Production of fungi.. 24

    2-3-2-1. Production of asexual reproduction.. 24

    2-3-2-2. Production of sexual reproduction.. 25

    2-3-3. Physiological structure of fungi.. 25

    2-3-4. The necessary conditions for the growth of mushrooms.. 26

    2-3-5. The mushrooms examined in this research. 27

    2-3-5-1. Microsporum genus (Microsporum. 27

    2-3-5-1-1. Microsporum canis (M. canis). 27

    2-3-5-1-2. Gypseum microsporum (M.gypseum. 28

    2-3-5-2. Genus 29. Trichophyton 2-3-5. 29. 2-3-5-2. Trichophyton rubrum. 30. 30

    2-3-5-3. Epidermophyton . 31

    2-3-5-3. Epidermophyton . 31

    2-3-5-4.

    2-3-5-4. Aspergillus fumigates. 32

    2-3-5-4. Aspergillus flavus (Aspergillus fumigates) 32

    2-3-5-5. Candida albicans. Fungal 2-4-1. Fungal infections. 2-4-2. Types of fungal diseases. 34. 2-4-2. Fungal disease of the external ear. 35. 2-4-2. Cutaneous fungal disease. 36.

    2-4-2-3. Aspergillosis .. 41

    2-4-2-3-1. Allergic aspergillosis.. 42

    2-4-2-3-2. Aspergilloma or colonized aspergillosis. 42

    2-4-2-3. Invasive aspergillosis.. 43

    2-4-2-4. Candidiasis.. 43

    2-4-2-4-1. Mucosal infections.. 44

    2-4-2-4-2. Skin diseases.. 44

    2-4-2-4-3. Systemic diseases. 44

    The fifth section: methods of checking antifungal agents

    2-5. Methods of measuring antifungal agents. 47

    2-5-1. Factors affecting the growth of microorganisms. 47

    2-5-1-1. Cultivation environment .. 47

    2-5-2-2. Effective factors in cultivation .. 47

    2-5-2-3. amount of mushrooms.. 48

    2-5-2-4. Temperature.. 48

    2-5-2.Oxygen.. 48

    2-5-3. Evaluation of antifungal compounds.. 49

    2-5-3-1. Dilution method.. 49

    2-5-3-1. Tube method.. 49

    2-5-3-2. Plate method using from the solid medium. 49

    2-5-3-1-3. Gradient plate method.. 50

    2-5-3-2. Turbidity measurement method.. 50

    2-5-3-3. Diffusion method.. 51

    2-5-3-3-1. Vertical diffusion.. 51

    2-5-3-3-1. Horizontal diffusion.. 51

    2-5-4. Factors Effective in measurement by diffusion method. 54

    2-5-4-1. Agar .. 54

    2-5-4-2. pH agar and solution to be measured. 55

    2-5-4-3. Cultivation conditions in the greenhouse. 56

    2-5-4-4. How to put microorganisms on Harvey Agar. 56

    2-5-4-5. The substance to be measured..Auras of growth and lack of growth. 57

    2-5-5. The accuracy of the results. 58

    2-5-6. Advantages and disadvantages of the publication method. 58

    2-5-7. Minimum inhibitory concentration. 59

    2-5-8. MIC determination methods. 59

    2-5-9. Effective factors in determining MIC. 59

     

     

    Chapter three: Materials and methods

     

    Part one: Preliminary works

     

    3-1-1. Materials and test equipment. 63

    3-1-2. Plant collection. 64

    3-1-3. to dry 64

    3-1-4. Powdering. 64

    3-1-5. Percolation. 64

    3-1-6. Concentration of extracts prepared by the device. 65

     

    Part Two: Mushroom Cultivation

     

    3-2-1. Preparation of required concentrations of plant extract. 67

    3-2-2. Preparation of culture medium. 67

    3-2-2-1. Well method. 67

    3-2-2-2. Mushroom cultivation. 68

    3-2-2-3. Method of mixing extract and culture medium. 68

    3-2-2-3. Preparation method of suspension of mushroom and plant extract. 68

    3-2-4. Control + . 68

    3-2-5. Control - . 68

    Chapter Four: Results

    1-4. Observation of plates after mushroom cultivation. 70

    Chapter Five: Discussion and conclusion

    1-5. Discussion and conclusion. 73

    2-5. Suggestions. 74

    English summary. 75

    List of sources. 76

    Attachments. 80

     

     

    Source:

     

    Zargari, A. Medicinal plants, Tehran university publication, fourth edition; 1990.p.1-4

    2-Ayatollahi S.A.M., Kobarfard F., Asgarpanah J., Choudhary M.I., Diterpenoids of Otostegia persica, Daru; 2009. p. 32-36

    3-Yassa, N[etal] cf., Otostegia persica as a source of natural antioxidants, Pharmaceutical Biology, 2005. 43(1), 33-38

    4-Ayatollahi S.A.M., Kobarfard F., Asgarpanah J., Choudhary M.I., 2010, Antiglycation activity of Otostegia persica, Afr.J.Biotech., 9(18), 43-46

    5- Ebrahimpoor MR, Khaksar Z., Noorafshan A. Antidiabetic effect of orally administered Otostegia persica extract on Streptozotocin diabetic rats, DOI 10.1007/s00580-010-1030-5, Comp. Clin. Pathol.

    6-Hajhashemi V., Asghari G., Karami-Saravi Z. Rabbani M. Effects of Otostegia persica on morphine withdrawal syndrome in mice, Int.J.Pharm. Res. 3, 2004. p. 171-175

    7-Asghri G., Nourollahi H., Havaie S.A., Issa L. Antimicrobial activity of Otostegia persica extracts, Res.Pharm.Sci.2004.p. 1, 53-58

    8- Khanavi, M. "Phytochemical investigation of Stachy". Thesis. Ermagnozi, University of Tehran - School of Pharmacy; 1373. p. 51-50

    9- Viqar Uddin A., et al. [Four new diterpenoids from Ballota limbata. Helvitica Chimica Acta.2004;87:682-89

    10-Gahreman A.‚Attra F :Biodiversity of Plant Species.Tehran University Publication.2005.p.101-110

    11-Zargari‚A.Medicinal Plant.Tehran University Publication;1989:Vol.2.P:2-5

    12-Gahreman A.Color Atlas of Iranian Flora.The Resaerch Institue of forest and ranglands-Botanical Department Publication; 1987.Vol.8.P:1731

    13-Shinwariz.k.‚KhanA.A.‚NakaikeT.Medicinal and other useful plants of District .Swat ‚Pakistan; 2003.P.82

    14-Watt G.Dictionary of the economic products of India.1972.P.336

     

    15-Asahi, M., [etal]. Purification and characterization of major extracellular proteinases from Trichophyton rubrum. Biochem. J. 1985,232:139-144

    16-Bishop,C.T.,Perry,M.B.,and Blank,F., The water-soluble polysaccharide of dermato-phytes.Can.J.Chem.1966.44:2291-2297.

    17-McGinnis,M.R.CurrenttopicsinmedicalMyology.Springer-Velag,New York,Berlin,Heidlberg,Tokyo.1985.p.54-61

    18-Beneke,E.S.Amd Rogers,A.L.,.Medical Myology and Human Mycosis.Star Publishing Company,Korea.1999.p.41-50

    19-Emami Massoud, Kord Paryoosh, Moghadami Mohin, Zaini Farideh 1373 Medical Mycosis, published Fourth, Tehran University Publications

    20-Mirzaei Masoumeh. Investigation of pulmonary fungal disease in tuberculosis and non-tuberculous patients and determining the identity of their agents. Tehran, Faculty of Health, Doctoral Thesis of Pharmacy, 1381, p. 52

    21-Barr, C.E.  and Torosian, J.P.

Investigating the antifungal effect of the total methanolic extract and etherdopterylic, chloroformic, ethyl acetate, methanolic and aqueous fractions of the flowering aerial parts of Otostegiapersica (Burm.) Boiss
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