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Pharmaceutical Sciences IAJPS 2018, 05 (05), 4766-4773 YAPI Adon Basile et al ISSN 2349-7750 CODEN [USA]: IAJPBB ISSN: 2349-7750 INDO AMERICAN JOURNAL OF PHARMACEUTICAL SCIENCES Available online at: http://www.iajps.com Research Article ETHNOBOTANICAL STUDY AND COMPARISON OF ANTITRICHOPHYTIC ACTIVITY LEAVES OF ASPILIA AFRICANA (PERS.) CD ADAMS VAR. AFRICANA, AGERATUM CONYZOIDES L. AND ACANTHOSPERMUM HISPIDUM DC. ON THE IN VITRO GROWTH OF TRICHOPHYTON MENTAGROPHYTES YAPI Adon Basile 1*, CAMARA Djeneb 1, COULIBALY Kiyinlma 2, ZIRIHI Guédé Noël 1 1Botanical Laboratory, UFR Biosciences, Félix HOUPHOUET BOIGNY University. PO Box, 582, Abidjan 22-Côte d’Ivoire. 2Biological Faculty of Sciences, Péléforo Gon Coulibaly University (Korhogo, Côte d’ivoire) PO Box, 1328 Korhogo Abstract : At the end of an ethnobotanical survey carried out in the district of Abidjan, Aspilia africana var africana, Ageratum conyzoides and Acanthospermum hispidum, three plant species widely known with weeds, were selected among the most used plants in the treatment of microbial diseases especially fungal ones. Thus, to make our contribution to the fight against opportunistic dermatophytosis in high recrudescence in the AIDS patients, we tested on Sabouraud medium the ethanolic and aqueous extracts of each of the three plants on the in vitro growth of a strain of Trichophyton mentagrophytes. The tests were carried out according to the method of double dilution in tilting tubes. The obtained results show that the tested T. mentagrophytes strain was sensitive to all the studied plant extracts. However, the EF70 %Ac extract has a better antifungal potential on T. mentagrophytes (MCF = 1,56 mg/mL and IC50 = 0,29 mg/mL). Otherwise, T. mentagrophytes was more sensitive to ethanolic extracts (EF70 % : MCF from 1,56 to 3,12 mg/mL and IC50 from 0,29 to 0,86 mg/mL) than aqueous extracts (ATE : MCF from 3,12 to 25 mg/mL and IC50 from 1,42 to 3,12 mg/mL ; ARF : MCF from 6,25 to 12,50 mg/mL and IC50 from 0,58 to 3,15 mg/mL). This work justifies the use in traditional environment of these weeds as anti-fungal ones. Thus, the 70% ethanolic extracts of A. africana, A. conyzoides and A. hispidum can serve as a basis for the development of phytomedicines against dermatophytosis. Keywords : Dermatophytosis, Aspilia africana, Ageratum conyzoides, Acanthospermum hispidum, Trichophyton mentagrophytes *Corresponding author : YAPI Adon Basile, QR code Botanical Laboratory, UFR Biosciences, Félix HOUPHOUET BOIGNY University. PO Box, 582, Abidjan 22-Côte d’Ivoire. E-Mail :[email protected] Please cite this article in press YAPI Adon Basile et al., Ethnobotanical Study and Comparison of Antitrichophytic activity Leaves of Aspilia Africana (Pers.) CD Adams Var. Africana, Ageratum Conyzoides l. And Acanthospermum Hispidum DC. On the In Vitro Growth of Trichophyton Mentagrophytes, Indo Am. J. P. Sci, 2018; 05(05). www.iajps.com Page 4766 IAJPS 2018, 05 (05), 4766-4773 YAPI Adon Basile et al ISSN 2349-7750 INTRODUCTION: MATERIAL AND METHODS The excessive increase in the prevalence of fungal Material infections over the last years, has profoundly Vegetal material transformed the focus on medical mycology. The plant material is made from the leaves of Aspilia According to the [1], fungal infections are africana (Pers.) C.D. Adams var africana, Ageratum responsible for more than 17 million deaths conyzoides L. and of Acanthospermum hispidum DC., worldwide each year, more than half of which come three Asteraceae collected in the district of Abidjan in from Africa alone. In Côte d'Ivoire, a study of August 2014. Their identification was performed at cutaneous mycosis revealed that 73.26 % of the National Floristic Centre (NFC) from the consultations in conventional medicine represented University Felix Houphouet-Boigny Abidjan-Cocody dermatological diseases, of which 61 % of cases are where samples are preserved. superficial mycosis [2]. Similarly, studies conducted Fungal germ from 1996 to 2000 also revealed that in all cases of The microbial strain is a fungal germ (Trichophyton dermatology consultations, 52.12 % of cutaneous mentagrophytes var. mentagrophytes) provided by mycosis were caused by yeasts and 44.78 % by the Mycology Laboratory at Training and Research dermatophytes. The latter are widespread filamentous Unit of the Faculty of Medical Sciences of Félix fungi that affect all social classes and have an affinity Houphouët-Boigny, University of Côte d'Ivoire. This for keratin. Depending on the location of the lesions mould was isolated from the nails of the large left toe on the skin, we can distinguish those of the glabrous of a patient with chronic urticaria (onyxis). skin, scalp ringworm and onyxis [3]. Cosmopolitan Culture medium affection, the dermatophytosis are first epidermal, Sabouraud medium (HIMEDIA/Ref : M1067-500G extensive, itchy and contagious. Heat and humidity, Lot 0000215703) was used for the culture of fungal immunosuppressive diseases, diabetes, promiscuity germ. with animals, hypersudation, obesity and certain drugs such as corticosteroids contribute to the Methods unexpected arrival dermatophytosis [4]. When they Monographic study of Aspilia africana, Ageratum are not treated, there is a multiplication and an conyzoides and Acanthospermum hispidum extension of the lesions. The evolution of To allow a better recognition of these three plants in a dermatophytosis is generally benign but chronic and natural environment, a complete and comparative the prognosis is always favorable. Among monographic study was carried out. It takes into dermatophytes, the genus Trichophyton is the most account the general appearance of the plants, the frequently implicated (71.55 % of isolates) [5]. The detailed description of the various organs as well as increase of this epidemiological modification is due some therapeutic uses of each of the plants. to several factors especially to the resistance of certain germs to the usual drugs [6,7,8]. Faced with Preparation of plant extracts this situation, several laboratories have undertaken The leaves of these three species were dried researches for the development of new drugs [9]. It is separately in the Laboratory for two weeks and within this framework that we undertook to evaluate reduced to a fine powder using an electric grinder in vitro and to compare the antitrichophytic activity type IKA Labortechnik (MFC type). of Aspilia africana (Pers.) C.D. var. africana, Preparation of aqueous total extracts (ATE) : the Ageratum conyzoides L. and Acanthospermum preparation of these extracts was performed using the hispidum DC., three medicinal plants with multiple method described by [8] which consists in macerating therapeutic properties selected at the end of an 100 g of plant powder of each species in 1L of sterile ethnobotanical survey in the district of Abidjan [10]. distilled water using a blender Blinder type 7 SEVEN Numerous pharmacological, phytochemical and STAR. The homogenates were filtered over toxicological studies was carried out by researchers hydrophilic cotton and then on filter paper Whatman to verify and justify the traditional use of these three 3 mm. The aqueous filtrate thus obtained are plants [11,12,13]. This study is part of the program of evaporated in an oven type Med Center Venticell at scientific valorization of the traditional 50 °C to obtain powders that constitute the aqueous pharmacopoeia of our laboratory. Its purpose is to extracts (ATEAa for Aspilia africana, ATEAc for search from our floristic inheritance, new Ageratum conyzoides and ATEAh for antimicrobial molecules to better control the Acanthospermum hispidum). chemotherapeutic approach against infectious Preparation of ethanolic fractions 70 % (EF70 %) diseases. and aqueous residual fractions (ARF) : these fractions were obtained separately by dissolving 5 g www.iajps.com Page 4767 IAJPS 2018, 05 (05), 4766-4773 YAPI Adon Basile et al ISSN 2349-7750 of each ATE in 100 mL of a ethanol 70 % solution Sterilization : the 12 tubes of each series were and then homogenized. After decantation and autoclaved (PBI STEMATIC III) at 121 °C for 15 filtration of the alcoholic fraction on hydrophilic min and then all the tubes were incubated at 30 °C for cotton and on filter paper Whatman 3 mm, the filtrate 10 days and then tilted with a small stick to the room collected is evaporated in an oven at 50 °C. The temperature to permit cooling and solidification of powder obtained constitutes the EF70 % extract the agar. (EF70 %Aa or EF70 %Ac or EF70 %Ah according to Colonies counting : after the incubation time, the the specy). Likewise the aqueous residual deposit was colonies of T. mentagrophytes were counted by direct collected and evaporated in an oven at 50 °C. The counting with a colonies counter pen type Geiger. powder obtained constitutes the ARF extract and is The growth in experimental tubes was evaluated as a called ARFAa or ARFAc or ARFAh according to ATE. percentage of survival, calculated at 100 % survival in the control growth control tube. The calculation of Yield calculation the percentage of survival was done according to the The yield is the amount of extract obtained from the following formula : plant powder. It is expressed as a percentage or S = (n / N) x 100 without any unit. In practice, it is determined by the (S = Survival of T. mentagrophytes percentage ; n = ratio of weight of the solids content after evaporation number of colonies in the control tube ; N = Number by the weight of the dry powder of the plant material of colonies in the test tube). used for the extraction, multiplied by 100. This gives Required antifungal parameters : data processing the following formula : has determined the following parameters antifungals : Yd = (m x 100) / M -MIC (Minimum Inhibitory Concentration) : this is (Yd : Extraction yield in percentage ; m : mass in the concentration of extract in the tube for which grams of the dry extract ; M : mass in grams of the there was no growth visible to the naked eye ; drug powder). -IC50 (Concentration for fifty percent inhibition) : is the concentration which gives 50 % inhibition.
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