in vitro phytochemical and biological studies of the hydroethanolic extract of Anchomanes difformis used in phytotherapy in Togo

in vitro phytochemical and biological studies of the hydroethanolic extract of Anchomanes difformis used in phytotherapy in Togo | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article in vitro phytochemical and biological studies of the hydroethanolic extract of Anchomanes difformis used in phytotherapy in Togo Tchilabalo Bouyo, Komi Koukoura Komi, Sandrine Tènè Salifou, Passimna Pissang, and 12 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7401969/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 28 Nov, 2025 Read the published version in Scientific Reports → Version 1 posted 13 You are reading this latest preprint version Abstract Background The proliferation of strains that are multi-resistant to conventional antibiotics has led to a need for research into other active molecules. The aim of this study was to evaluate the antimicrobial activity of Anchomanes difformis , a medicinal plant from the West African sub-region used in traditional pharmacopoeia. Methods Total hydroethanolic extracts of the different organs (leaves, bulb and rhizomes) were obtained by maceration. Qualitative phytochemistry of the extracts was carried out using staining methods to identify the main chemical groups. Total flavonoids were quantified by spectrophotometric assay. Two methods were used to assess antioxidant activity: the phosphomobyldate reduction method and the FRAP test. in vitro antimicrobial tests were carried out using the solid-state diffusion method (Puits et disque). Results The highest hydroethanol extraction yield was obtained with the A. difformis leaf bark extract (9.47%). The results showed the presence of alkaloids, coumarins and reducing sugars. The highest content of total flavonoids was found in the bulbs, with 33.78 µg Eq routine/mg DM. The plant leaves showed high antioxidant activity in vitro with 37.79 EAA/g DM and 270.50 µmol Eq FeSO4/mg DM. On completion of the antimicrobial tests, there was no visible inhibition zone diameter for all the extracts with the two methods used. Conclusion The present study invalidates the local use of this plant in pathologies caused by the microbial strains studied. However, research into other activities associated with this plant could explain its use in traditional medicine. Biological sciences/Biochemistry Biological sciences/Biological techniques Biological sciences/Drug discovery Biological sciences/Microbiology Biological sciences/Plant sciences Anchomanes difformis phytochemistry activity antioxidant antimicrobial Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Introduction Antibiotic resistance occurs when bacteria develop the ability to evade therapeutic molecules [1,2]. Today, the growing resistance of pathogens to antibiotics is a major global health threat [3,4]. In West Africa, the endemicity of respiratory infections, bacterial meningitis, diarrhoea and other infectious diseases has increased the use of antibiotics both for symptomatic treatment and prophylaxis [5]. This is eroding the effectiveness of conventional treatments and forcing the urgent discovery of new antimicrobial compounds [6]. In this context, medicinal plants are attracting renewed interest: rich in various secondary metabolites, they represent one of the most promising sources of innovative bioactive molecules [4,7,8]. Numerous studies point out that the plant kingdom produces a wide variety of compounds (alcohols, terpenes, alkaloids, flavonoids, etc.), some of which have already served as models for the development of new drugs [4,7]. At the same time, herbal medicine plays a central role in traditional health care in many countries. According to the World Health Organization, about 80% of the population in some developing countries relies on herbal medicines for their primary care [9,10]. This is especially true in sub-Saharan Africa, where the accessibility and low cost of herbal remedies make them an essential resource. This practical importance goes hand in hand with the need to renew the therapeutic reservoir: the discovery of new molecules of plant origin, with antibacterial, antioxidant or anti-inflammatory activities, has become a major challenge in pharmaceutical research [6,7]. In Togo, traditional knowledge about medicinal plants continues to play an important role in primary health care [11]. In this general framework, Anchomanes difformis (Blume) Engl. (family Araceae) appears to be a plant of interest. Anchomanes difformis , locally called "Adodo" in Togo, is a medicinal plant used in traditional African medicine, particularly in Nigeria, Togo and Côte d'Ivoire [12,13]. It grows mainly in humid tropical areas, savannahs and along waterways at low or medium altitudes [12,13]. This species is a perennial herbaceous plant, with a horizontal tuber measuring between 40–80 cm long and 8–22 cm in diameter, with annular scars [12,13]. Each season, it produces a single large tripartite leaf, up to 1.5 m wide, with sessile leaflets of variable shape [12,13]. The petiole, greenish purple, reaches 2.5 m in length and bears small green needles [12,13]. The stem is green with a white base [12,13]. The inflorescence emerges from the tuber, with a cylindrical spadix bearing female flowers with purple-pink ovary at the base, followed by creamy-white male flowers [12,13]. It is widely used in traditional local medicine: roots, rhizomes and leaves are used against an impressive variety of ailments (abdominal pain, respiratory disorders, joint problems, infections, digestive disorders, neurological disorders, etc.) [12,13]. The roots of A. difformis are used in Benin to treat anal wounds, oral wounds, dysentery, diabetes and its complications and dysentery [14]. Other surveys have shown that all parts of the plant treat asthma [15–17], malaria [18], cough and throat-related problems [13]. Some investigations have reported the use of powdered root mixed with palm oil is used as a remedy for respiratory diseases in children in Zaire [19]. The antimicrobial properties of the plant have been reported [16,20]. It is cited as a diuretic and "all-comer" remedy in the Togolese and Nigerian pharmacopoeia [12,13]. However, despite its widespread traditional use, the available scientific data on this species remain limited. As noted in recent work, very few studies have characterized the chemical composition or assessed the biological activities of A. difformis [21–26]. The lack of valid data makes a rigorous review of its active ingredients even more relevant. In this perspective, the present study aims to fill these gaps with a systematic approach: it aims to analyze the phytochemical composition of the hydroethanolic extract of Anchomanes difformis and to evaluate its potential in vitro biological activities (in antioxidant and antimicrobial). The aim is to provide a scientific basis for the traditional uses of this plant and to search for new therapeutic agents of plant origin. This approach is in line with the current need to discover, within plant biodiversity, molecules capable of effectively fighting antibiotic-resistant pathogens [4,6]. Materials and methods Collection of plant material Leaves (Fig. 1 ), rhizomes and bulbs (Fig. 2 ) of Anchomanes difformis were collected in Dalavé (Tsévié, Maritime Region of Togo) in October 2021. The material plants have been botanically authenticated at the Herbarium of the Department of Botany, Faculty of Sciences, University of Lome. Bacterial strains The microorganisms used for the test were clinical and reference strains of Streptococcus pneumoniae , Escherichia coli , Staphylococcus aureus and Candida albicans , all isolated at the National Institute of Hygiene in Lomé (INH-Lomé). Extraction The extraction was carried out in accordance with[27] and taken up by [28]. The samples of each plant were carefully washed under running water and dried in the open air at room temperature for two weeks. After drying, the plant materials were reduced to powder using the grinding machine. The ethanolic extraction was carried out by macerating 100 g of powder in 1,000 ml of 70% ethanol (v/v), under continuous stirring for 48 hours at room temperature. The extract was filtered through WHATMAN N°1 filter paper and evaporated at 45°C using a rotary evaporator until dryness under reduced pressure. The extracts were stored at 4°C in the refrigerator until used. Phytochemical analysis Phytochemical screening was performed based on staining characteristic tests to highlight major chemical groups. It focused on the hydroethanolic extracts of the plants studied. The chemical groups were identified by reference to the methods described by [29] and taken up by [30–32]. Alkaloids The dissolved extracts were divided into two tubes at a rate of 2 mL each. In a first tube (test tube), 4 mL of sulphuric acid (H 2 SO 4 ) diluted to 10% and a few drops of Dragendorff's reagent were added. The appearance of an orange-red precipitate indicates the presence of alkaloids in the hydroethanolic extract. The second tube served as a control. Flavonoids The dissolved extracts were divided into two tubes at a rate of 2 mL each. In a first tube (test tube), 4 mL of methanol and a few drops of magnesium turns were added; then 1 mL of concentrated HCl was added. The appearance of a red coloration indicates the presence of flavonoids in the hydroethanolic extract. The second tube served as a control. Saponines The dissolved extracts were divided into two tubes at a rate of 2 mL each. In a first tube (test tube), a few drops of distilled water were added. The appearance and persistence of foam after stirring indicates the presence of saponosides. The second tube served as a control. Tannins The dissolved extracts were divided into two tubes at a rate of 4 mL each. In a first tube (test tube), a few drops of FeCl 3 were added. The appearance of a blackish brown colour after homogenization of the mixture indicates the presence of tannins in the hydroethanolic extract. The second tube served as a control. Phenols The dissolved extracts were divided into two tubes at a rate of 4 mL each. In a first tube (test tube), a few drops of FeCl 3 were added. The appearance of a blackish brown colour after homogenization of the mixture indicates the presence of phenolic compounds in the hydroethanolic extract. Triterpenes and sterols The dissolved extracts were divided into two tubes at a rate of 4 mL each. In a first tube (test tube), 1.6 mL of chloroform and 2.6 mL of sulphuric acid (H 2 SO 4 ) were added. The appearance of a reddish-brown ring between two phases, one clear at the bottom and the other green at the top, indicates the presence of triterpenes and sterols in the hydroethanolic extract. The second tube served as a control. Coumarins The dissolved extracts were divided into two tubes at a rate of 4 mL each. In a first tube (test tube), 2 mL of distilled water was added, and a small amount of ammonia was added to the mixture. The appearance of fluorescence after ultraviolet illumination indicates the presence of coumarins in the hydroethanolic extract. The second tube served as a control. Reducing sugars The dissolved extracts were divided into two tubes at a rate of 2 mL each. In a first tube (test tube), 2 mL of Fehling's Liquor (1 mL of solution A + 1 mL of solution B) were added and brought to a boil in a boiling sea bath for 2 to 3 min. The appearance of a brick-red precipitate indicates the presence of reducing sugars in the hydroethanolic extract. The second tube served as a control. Flavonoid content The flavonoid content is estimated according to the method described by [33] and taken up by [30–32]. It consisted of preparing plant extracts at 1mg/mL in distilled water, aluminum chloride (AlCl 3 ) 2% in distilled water, and rutin at different concentrations of 0; 5 ; 25 ; 50 ; 75 ; 100 ; 150 ; 200 µg/mL in methanol. The operation consisted of mixing 1 mL of the extract solution at 1 mg/ml or 1 mL of each rutin concentration with 1 mL of aluminum chloride (AlCl 3 ) 2% using a vortex. After 10 minutes of incubation, the absorbance was directly measured with a UV-visible spectrophotometer (METASH UV-5200PC UV/VIS Spectrophotometer) at 415 nm against a blank. Rutin was used as a standard. The total flavonoid content of the extracts was inferred from the calibration curve established with Rutin (0–200 µg/mL) and the results were expressed in microgram equivalent of rutin per milligram of dry matter (µg ER/mg DM). Three tests were conducted for each extract. The total flavonoid content ( XFlav ) is obtained using the following formula: XFlav Total flavonoid content (µg ER/mg DM) OD Optical Density read at 415 nm Antioxidant activity Phosphomobyldate reduction test Determination of antioxidant activity by the phosphomobyldate reduction assay was performed using the method described by [34] and slightly modified in [31,32]. The phosphomolybdate reagent was prepared (100 mL of reagent) from a mixture of 90 mL of 0.6 M sulphuric acid, 5 mL of 0.1% sodium phosphate and 5 mL of 1% ammonium molybdate. For the test, 1 mL of each extract was added to 9 mL of the above reagent. The whole thing was brought to a temperature of 95°C in a water bath for 90 minutes. Then the mixture was cooled to room temperature. Absorbances were measured at wavelengths of 820 nm against a blank consisting of reagent and distilled water. Ascorbic acid was used as the standard antioxidant under the same operating conditions and the results were expressed in milligrams of ascorbic acid equivalent per gram of dry matter (mg AAE/g DM). Three tests were performed for each concentration of product tested. The antioxidant activity related to the reducing power of the extracts is expressed in Antioxidant Content (X Mob ) using the following formula: XMob Antioxidant Content (mg AAE/g DM) OD Optical Density read at 820 nm Test FRAP (Ferric Reducing antioxidant Power) The determination of antioxidant activity by the FRAP assay was carried out using the method described by [35] and taken up by [30–32,36]. Thus, 3 mL of the freshly prepared FRAP test reagent in a test tube, 100 µL of the various solutions of iron II sulphate with concentrations between 0 and 2000 µmol. L − 1 will be added. The mixture was vigorously shaken at the vortex and the optical density was read after 5 min with a spectrophotometer at 593 nm. The absorbance of the TPTZ-Fe 2+ complex allowed to draw a calibration curve from the concentration range (0–2000 µM) of the iron sulfate solution (FeSO 4 , 7H 2 O) dissolved in methanol. For the samples of the extracts to be tested, the FRAP reagent (3 mL) and the solution of the extract to be tested (100 µL) with a strength of 1 mg/mL were mixed in the same proportions as for the standard curve plotting. The optical density was read after 5 min at 593 nm. The antioxidant capacity of the extracts will be measured using the calibration curve by the color change related to the formation of the complex (Fe 2+ TPTZ) and expressed in micromole equivalent of iron sulfate per milligram of dry matter (µmol Eq FeS0 4 /mg DM). The tests will be repeated 3 times. The antioxidant activity related to the reducing power of the extracts is expressed in Reducing Power (X FRAP) using the following formula: XFRAP Reducing power (µmol Eq FeS0 4 /mg DM) OD optical density read at 593 nm Evaluation of the antimicrobial potency of extracts The identification of the active extracts was carried out by the solid-state diffusion method [37] and the determination of the minimum inhibitory concentration (MIC) by the liquid dilution method [38]. Preparation of microbial strains After collection, the strains were transplanted to their appropriate media (Chapman for S. aureus , Sabouraud + Chloramphenicol for C. albicans , Mac Conkey for E. coli and Fresh Blood Agar (FSG) for S. pneumoniae ) and stored. To obtain the 24-hour colonies that were used to prepare the inoculum, the strains were transplanted on agar medium without inhibitors [39]. Preparation of the inoculum From an 18–24 h culture on agar medium, a suspension was prepared in saline solution (0.9% NaCl) equivalent to the Mac Farland standard 0.5 (~ 10 8 CFU/ml). Inoculation was sown by flooding with the inoculum suspension diluted to 1/100 (~ 10 6 CFU/ml) in accordance with the necessary safety measures [39]. Preparation and sterilization of the stock solution of extracts The extracts were dissolved in sterile distilled water to an initial concentration of 200 mg/mL (200 mg of each extract in 1 mL of distilled water). To verify their sterility, an aliquot of each stock solution was taken and inoculated on Muller Hinton agar. After incubation at 37 °C for 24 hours, those that gave colonies were filtered with a millipore membrane of 0.45 µm in diameter. After sterilization, each stock solution is diluted 1/4 with distilled water to obtain solutions with a concentration of 50 mg/mL of each extract. In vitro antimicrobial activity Antimicrobial testing of extracts on germs tested by the well method Mueller Hinton (MH) agar plates for bacteria and Sabouraud dextrose + Chloramphenicol for yeasts were inoculated by flooding following the recommendations of the Antibiogram Committee of the French Society of Microbiology [39]. After the dishes have dried, (about 5 min) the agar is perforated into a maximum of six wells (90 mm Petri dishes) with a sterile tip previously cut so that the diameter is about 6 mm. The cavities formed are filled with 50 µL of the 50 mg/mL extract solution (50 µL per well). The boxes are placed at room temperature 25°C for 15 minutes for a pre-diffusion phase. Incubated at 37 °C for 24 hours for bacteria and 48 hours for yeasts. Ciprofloxacin (bacteria), Nystatin (yeast) were used as a positive control and distilled water as a negative control. After 24 hours of incubation, the diameters of the inhibition zones were measured using a graduated ruler. The tests are carried out three times and an average are obtained [37]. MICs and BMCs were determined for extracts with an inhibition diameter greater than or equal to 11 mm. Antimicrobial testing of extracts on germs tested by the disc method Mueller Hinton (MH) agar plates for bacteria and Sabouraud dextrose + Chloramphenicol for yeasts is dried (about 5 min) and inoculated by flooding according to the recommendations of the Antibiogram Committee of the French Society of Microbiology [39]. Disc preparation WATTMAN (No. 1) 6 mm diameter, virgin, autoclaved paper was impregnated with 50 µL of the extracts at the initial concentration of 50 mg/mL. The final load of the disc is 2.5 mg. WATTMAN N°1 papers are dried in an oven (37 °C ). Sensitivity test These discs are placed aseptically on the previously inoculated agar. The boxes are placed at room temperature 25 °C for 15 minutes for a pre-diffusion phase. Incubated at 37 °C for 24 hours for bacteria and 48 hours for yeasts. Ciprofloxacin (bacteria), Nystatin (yeast) were used as a positive control and distilled water as a negative control. After 24 hours of incubation, the inhibition diameters were measured. The tests are carried out three times and an average are obtained on the three determinations [37]. MICs and BMCs were determined for extracts with an inhibition diameter greater than or equal to 11 mm. Determination of the minimum inhibitory concentration (MIC) The minimum inhibitory concentration (MIC) was determined by the microplate microdilution method [38]. Mueller Hinton Broth is used to prepare serial dilutions at half concentrations ranging from 50 to 0.097 mg/mL. The inoculum (10 µL) was added to each cup containing 50 µL of extract. Cupules without inoculum were considered as negative controls. All these cups were incubated at 37°C for 24 h. The first cup in the series showing no sign of culture visible to the naked eye was MIC. An aliquot of the contents of each cupule with no visible growth was taken and then seeded by spreading on Mueller Hinton medium [38]. The dishes are incubated at 37°C for 24 to 48 hours at the end of which the number of colonies is determined. After incubation, the smallest concentration of the extract that does not produce colonies is considered the MBC. To determine whether the antimicrobial effect observed is bactericidal or bacteriostatic, the MBC/CMI ratio is used to assess the activity of the extracts (bactericidal/bacteriostatic). For a CMB/MIC ratio greater than 1, the antimicrobial effect is bacteriostatic.[40] For a CMB/MIC ratio of 1, the antimicrobial effect is bactericidal [40]. Statistical analysis of the data The data obtained was subjected to statistical analysis using RStidio 4.1.3 software from 10/03/2022 and Microsoft® Excel® spreadsheet for Microsoft 365 MSO 2021. Quantitative variables (flavonoid content, FRAP test, and phosphomobyldate reduction method) are presented as a mean and standard deviation. One-Factor Analysis of Variance (ANOVA) was used to assess the diameters of the zones of inhibition. The significance level has been set at 5%. Results For this study, the best yield is obtained by the extract of the leaves of A. difformis (9.47%). The weakest is obtained with the extract of the rhizomes of A. difformis (4.10%). All extracts have an acidic pH (pH ˂ 7) and range from 6.28 to 5.40. The solubility is quite good, compared to the solvent used for all the extracts. The colour of the extracts is light yellow with a sticky gum appearance for all three extracts (Table 1 ). The extracts were analyzed for alkaloids, tannins, flavonoids, saponosides, reducing sugars, triterpenoids, and sterols. The result is recorded in Table 2 . Hydroethanolic extracts of leaves, rhizomes and bulbs of A. difformis contain only alkaloids, coumarins and reducing sugars (Table 2 ). Quantitative tests show a low presence of flavonoids in all extracts at various doses (Table 3 ). The highest flavonoid content was measured with the hydroethanolic extract of the bulb of A. difformis (33.78 µg ER/mg DM) and the lowest with the hydroethanolic extract of the leaves of A. difformis (28.85 µg ER/mg DM). The evaluation of antioxidant activity revealed that all extracts have antioxidant power. The best antioxidant activity was observed with A. difformis leaf extract with 37.79 mg AAE/g DM by the phosphomolybdate reduction assay and 270.50 µmol FeS0 4 /mg DM by the FRAP assay (Table 4 ). Regarding antimicrobial activity, at the initial concentration of 50 mg/ml, the three hydroethanolic extracts tested in this work did not significantly inhibit the growth of microorganisms (p = 2.e-16, cv = 3.862–5.562%) with the two diffusion methods on all strains used (Tables 5 & 6 ). Therefore, the MIC was not determined (Inhibition Diameter ˂11 mm). Table 1 Some characteristics of the extracts obtained Plants Organs Aspect Colour Solubility pH Yield A. difformis Leaves Sg Ly Pg 5,40 9,47% Bulbs Sg Ly Pg 6,28 4,14% Rhizomes Sg Ly Pg 5,65 4,10% Sg: Sticky Gum; Ly: Light yellow; pH: Hydrogen potential; Pg: Pretty good Table 2 Phytochemical study of the studied extracts Secondary metabolites Extraits HE of A. difformis Leaves Bulbs Rhizomes Alkaloids + + + Flavonoids - - - Tannins - - - Coumarins + + + Saponosides - - - Triterpenes - - - Phenol - - - Sugar Reducers + + + Legend: -: Absence; +: Presence; HE: Hydroethanolic Table 3 Flavonoid content of the extracts studied HE Extracts A. difformis Leaves Bulbs Rhizomes Total flavonoid content (µg ER/mg DM) 24,85 ± 0,26 33,78 ± 0,66 29,82 ± 0,88 ER: Equivalent to Rutine; HE: Hydroethanolic; DM: Dry matter Table 4 Antioxidant activities in vitro Excerpts studied Hydroethanolic extracts Phosphomolybdate Reduction Test (mg AAE/g DM) FRAP Testing (µmol FeS0 4 /mg DM) Feuilles de A. difformis 37,79 ± 0,18 270,50 ± 5,00 Bulbs of A. difformis 19,59 ± 0,00 202,17 ± 2,89 Rhizomes of A. difformis 23,09 ± 0,20 190,50 ± 0,00 AAE: Acide Ascorbique Equivalent ; FRAP: Ferric Reducing antioxidant Power ; DM: Dry matter Antimicrobial activities Table 5 Antimicrobial testing result of extracts on germs tested by The well method Microbial strains Extraits HE of A. difformis Water-Ethan NC Cyprus/Nyst. PC (50 µg/mL) P-value CV(%) Leaves (50 mg/mL) Bulbs (50 mg/mL) Rhizomes (50 mg/mL) S. aureus ATCC 29213 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6.00 ± 0.00 b 37.67 ± 0.57 a 2 e−16 *** 3,856 S. aureus 0689 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6.00 ± 0.00 b 25.67 ± 0.57 a 2 e−16 *** 3,833 E. coli ATCC 25922 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 44.00 ± 1.00 a 2 e−16 *** 4,225 E. coli 1628 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6.00 ± 0.00 b 17.00 ± 1.00 a 2 e−16 *** 4,037 S. pneumonia ATCC 49619 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 42.33 ± 0.57 a 2 e−16 *** 3,922 S. pneumonia 034 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 33.33 ± 0.57 a 2 e−16 *** 3,362 C. albicans ATCC 10231 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 32.00 ± 0.57 a 2 e−16 *** 4,495 C. albicans 1134 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 29.33 ± 0.57 a 2 e−16 *** 5.562 The values are expressed as the Mean ± Standard Error. Values in the same column followed by the same lowercase letter are statistically identical (Duncan, p ˂ 0.05) HE: Hydroethanolic; NC: Negative control; PC: positive control; CV: Coefficient of Variance Table 6 Result of antimicrobial testing of extracts on germs tested by the disc method Microbial strains Extraits HE of A. difformis Water-Ethan NC Cyprus/Nyst. PC (50 µg/mL) P-value CV(%) Leaves (50 mg/mL) Bulbs (50 mg/mL) Rhizomes (50 mg/mL) S. aureus ATCC 29213 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 38.33 ± 0.57 a 2 e−16 *** 2,91 S. aureus 0689 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 25.33 ± 0.57 a 2 e−16 *** 5,27 E. coli ATCC 25922 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 44.00 ± 1.00 a 2 e−16 *** 5,95 E. coli 1628 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 18.00 ± 1.00 a 1.8 e−16 *** 6,92 S. pneumonia ATCC 49619 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 42.33 ± 0.57 a 2 e−16 *** 5,05 S. pneumonia 034 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 32.67 ± 0.57 a 2 e−16 *** 5,80 C. albicans ATCC 10231 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 32.33 ± 0.57 a 2 e−16 *** 11,81 C. albicans 1134 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 6,00 ± 0,00 b 31.00 ± 1.00 a 2 e−16 *** 6.06 The values are expressed as the Mean ± Standard Error. Values in the same column followed by the same lowercase letter are statistically identical (Duncan, p ˂ 0.05) HE: Hydroethanolic; NC: Negative control; PC: positive control; CV: Coefficient of Variance Discussion The study evaluated the antimicrobial activities of A. difformis on clinical strains of E. coli , S. aureus , S. pneumoniae , and C. albicans . The choice of this plant as study material is guided by their common use in traditional medicine in Africa to cure various diseases.[41] - Yield Our results show that for this study, the best yield is obtained by the extract of A. difformis leaves (9.47%). The lowest is obtained with the extract of the rhizomes of A. difformis (4.10%). These low yields could be explained by the short time (48 hours) and especially the mode and speed of rotation to homogenize the solvent and plant powder mixture[42]. Hydroethanolic extraction is valued because of its ability to extract a wide range of compounds, its relative safety of use, and its low cost compared to other extraction methods.[43] - Qualitative and quantitative phytochemical study The qualitative phytochemical study of the three extracts of A. difformis revealed the presence of alkaloids, coumarins and reducing sugars. These results do not corroborate those described by [15,44] who, in addition to the alkaloids obtained, also found tannins, flavonoids, saponosides and phenols but not reducing sugars. This could be explained by the extraction method used. Our results showed a low presence of flavonoids in all extracts at various doses (Table 3 ). This result confirms the absence of flavonoids in A. difformis extracts in qualitative tests (Table 2 ). The lowest is that of the hydroethanolic extract of the leaves of A. difformis (28.85 µg ER/mg DM). Our results corroborate with those of [17,21], who also found that the flavonoid content of A. difformis leaves is lower with 4 mg ER/g DM using the same method. - Antioxidant activity Regarding antioxidant activity, the FRAP test and the phosphomobyldate reduction test gave a better reducing power of A. difformis leaves with 270.5 µmol FeS0 4 /mg DM and 37.79 mg AAE/g DM respectively. The low presence of flavonoids and other secondary metabolites could explain this low antioxidant activity of the various extracts of A. difformis [45]. - Antimicrobial activity Presumptive testing by the agar diffusion method (Well and Disc) revealed that the three extracts did not significantly inhibit the growth of microorganisms (p = 2.e − 16 , cv = 3.862–5.562%) on all bacterial strains used for the study. Our results are in contradiction with the work of[12] which has shown that the essential oil extract of A. difformis has antimicrobial activities on S. aureus and C. albicans . This could be explained by the difference in the nature of the extracts used in the two studies. The low presence of phytochemicals is thought to be the basis for the antimicrobial inactivities observed with A. difformis extracts [31]. Conclusion The objective of our study was to evaluate the biological and phytochemical activities of A. difformis on strains of E. coli , S. aureus , S. pneumoniae and C. albicans . Qualitative phytochemicals have shown that the extracts (leaves, rhizome and bulb) of A. difformis all contain coumarins, alkaloids and reducing sugars. Quantitative phytochemistry gave low total flavonoid content for all A. difformis extracts in vitro . Our results showed that the three hydroethanolic extracts of A. difformis did not significantly inhibit the growth of the microorganisms tested in this study. Ultimately, we can conclude that the hydroethanolic extracts of the leaves, rhizomes and bulbs of Anchomanes difformis would be inactive on all the germs tested ( E. coli , S. aureus , S. pneumoniae and C. albicans ). This study sheds light on the chemical components and biological activities of the hydroethanolic extract of Anchomanes difformis used in herbal medicine in Togo. Our results suggest that this plant could be a valuable source of bioactive compounds with potential applications in the treatment of various conditions other than the strain-related conditions tested in our study. Abbreviations ATCC: American Typographic Culture Collection ; MIC: Minimum Inhibitory Concentration ; MBC: Minimum Bactericidal Concentration ; CS NDE: Sisters of Our Lady of the Church Health Center ; INH: National Institute of Hygiene of Lomé ; MH: Mueller-Hinton ; WHO: World Health Organization Declarations Ethics Approval and Consent to Participate Not Applicable Consent to Publication Not Applicable Clinical trial number Not applicable Availability of Data and Materials The datasets used and/or analyzed in the current study are available from the corresponding author upon reasonable request. Competing Interests The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Authors' contributions Tchilabalo Bouyo, Sandrine Tènè Salifou, Jules Koffi Kpatagnon, Passimna Pissang and Komi Koukoura Komi harvested and identified the plant parts used. Tchilabalo Bouyo, Sandrine Tènè Salifou, Kodjovi Sossou, Samadou Tchakondo, Passimna Pissang, Yao Hoekou, Holaly Efui Gbékley and Komi Koukoura Komi participated in the extraction and flavonoid assay. Antioxidant activity tests were carried out by Tchilabalo Bouyo, Bawimodom Bidjada, Sandrine Tènè Salifou, Passimna Pissang, Jules Koffi Kpatagnon, Samadou Tchakondo, Abdoul Kader Ouedraogo, Isidore Kodjovi Anani Gbenonssi, Kodjovi Sossou, Komlan Tchalla, Yao Hoekou and Holaly Efui Gbékley. The results were analyzed statistically by Tchilabalo Bouyo, Holaly Efui Gbékley, Blaise Etienne M'BOUMBA and Passimna Pissang. Finally, Tchilabalo Bouyo, Richard Kouyassa Dessougmba, Passimna Pissang, Komi Koukoura Komi wrote the maniscrit and Komi Koukoura Komi and Tchadjobo Tchacondo approved and supervised the work. Funding declaration Lack of funding for this study. Acknowledgements The authors express their sincere thanks and deep gratitude to all those who have contributed to the realization of this work. Our thanks to the National Institute of Hygiene of Lomé (INH-Lomé) who provided us with the strains for the experiment and to the health center of the Sisters of Our Lady of the Church of Tokoin Seminary of Lomé (CS NDE) for their moral support throughout the study process. References Landecker H. Antibiotic Resistance and the Biology of History. Body Soc. 2016 Dec 1;22(4):19–52. Landecker, Hannah. La résistance aux antibiotiques et la biologie de l’Histoire. http://journals.openedition.org/rac [Internet]. 2021 Sep 1 [cited 2022 Dec 8];15(15–3). Available from: http://journals.openedition.org/rac/22123 Bouyahya A, Abrini J, Bakri Y, Dakka N. Screening phytochimique et évaluation de l’activité antioxydante et antibactérienne des extraits d’Origanum compactum. Phytotherapie. 2017;15(6). 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2022)\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7401969/v1/4d6482bb7dfd455a19a1a87f.png"},{"id":89891699,"identity":"94cfb2e0-95cc-4b1e-966b-455593abb27a","added_by":"auto","created_at":"2025-08-26 07:40:16","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":618468,"visible":true,"origin":"","legend":"\u003cp\u003eBulbs and rhizomes of Anchomanes difformis (Source: Nikon Coolpix P300, 2022)\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7401969/v1/b41078271dd19993a4b21a8b.png"},{"id":89891669,"identity":"63e263c5-6d44-4100-ad9b-7e205372b6cd","added_by":"auto","created_at":"2025-08-26 07:40:15","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":122660,"visible":true,"origin":"","legend":"\u003cp\u003eP extraction process on the métal plate (Source: Nikon Coolpix P300, 2022)\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-7401969/v1/e1b0694dc2de305f2b8fc414.png"},{"id":89892109,"identity":"9bbd9743-5efd-4e4b-8f39-26af33f21431","added_by":"auto","created_at":"2025-08-26 07:48:16","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":42478,"visible":true,"origin":"","legend":"\u003cp\u003eWHATMAN N°1 paper filtration process\u003c/p\u003e\n\u003cp\u003e(Source: Nikon Coolpix P300, 2022)\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-7401969/v1/50e19471ef16d4f6bbe59632.png"},{"id":89891692,"identity":"55d002d2-e10d-405f-8d16-68193be2c0ef","added_by":"auto","created_at":"2025-08-26 07:40:16","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":277974,"visible":true,"origin":"","legend":"\u003cp\u003eImage showing some qualitative test results of the phytochemical analysis\u003c/p\u003e\n\u003cp\u003e(Source: Nikon Coolpix P300, 2022)\u003c/p\u003e","description":"","filename":"floatimage7.png","url":"https://assets-eu.researchsquare.com/files/rs-7401969/v1/2ce0af086fb030d80739a20a.png"},{"id":89891663,"identity":"89893b63-4552-4b25-bb70-92225883aa01","added_by":"auto","created_at":"2025-08-26 07:40:15","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":173430,"visible":true,"origin":"","legend":"\u003cp\u003eWell Method Presumptive Antimicrobial Testing Process\u003c/p\u003e\n\u003cp\u003e(Source: Nikon Coolpix P300, 2022).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ea\u003c/strong\u003e: Digging wells on the sterile HD medium\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eb\u003c/strong\u003e: the introduction of plant extracts into the wells on the MH medium already sown by flooding\u003c/p\u003e","description":"","filename":"floatimage11.png","url":"https://assets-eu.researchsquare.com/files/rs-7401969/v1/6eddc9b5f48709de3ab5de16.png"},{"id":89892107,"identity":"0489cee1-e2c5-4933-8d5c-4793fe76a21c","added_by":"auto","created_at":"2025-08-26 07:48:16","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":173571,"visible":true,"origin":"","legend":"\u003cp\u003eDisc Method Presumptive Antimicrobial Testing Process\u003c/p\u003e\n\u003cp\u003e(Source: Nikon Coolpix P300, 2022)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ea\u003c/strong\u003e: Design of the discs using WHATMAN N°1 paper\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eb\u003c/strong\u003e: Deposits of the extract-impregnated discs on the MH medium already inoculated by flooding with bacteria\u003c/p\u003e","description":"","filename":"floatimage13.png","url":"https://assets-eu.researchsquare.com/files/rs-7401969/v1/8beb90fe38b04bc042c998e8.png"},{"id":97178779,"identity":"676dc4ab-5561-48c0-aae3-8dd56ad5648b","added_by":"auto","created_at":"2025-12-01 16:13:37","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3751078,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7401969/v1/7ada0b2e-c72b-465b-80f4-9487c3bcc195.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"in vitro phytochemical and biological studies of the hydroethanolic extract of Anchomanes difformis used in phytotherapy in Togo","fulltext":[{"header":"Introduction","content":"\u003cp\u003eAntibiotic resistance occurs when bacteria develop the ability to evade therapeutic molecules [1,2]. Today, the growing resistance of pathogens to antibiotics is a major global health threat [3,4]. In West Africa, the endemicity of respiratory infections, bacterial meningitis, diarrhoea and other infectious diseases has increased the use of antibiotics both for symptomatic treatment and prophylaxis [5]. This is eroding the effectiveness of conventional treatments and forcing the urgent discovery of new antimicrobial compounds [6].\u003c/p\u003e\u003cp\u003eIn this context, medicinal plants are attracting renewed interest: rich in various secondary metabolites, they represent one of the most promising sources of innovative bioactive molecules [4,7,8]. Numerous studies point out that the plant kingdom produces a wide variety of compounds (alcohols, terpenes, alkaloids, flavonoids, etc.), some of which have already served as models for the development of new drugs [4,7].\u003c/p\u003e\u003cp\u003eAt the same time, herbal medicine plays a central role in traditional health care in many countries. According to the World Health Organization, about 80% of the population in some developing countries relies on herbal medicines for their primary care [9,10]. This is especially true in sub-Saharan Africa, where the accessibility and low cost of herbal remedies make them an essential resource. This practical importance goes hand in hand with the need to renew the therapeutic reservoir: the discovery of new molecules of plant origin, with antibacterial, antioxidant or anti-inflammatory activities, has become a major challenge in pharmaceutical research [6,7]. In Togo, traditional knowledge about medicinal plants continues to play an important role in primary health care [11].\u003c/p\u003e\u003cp\u003eIn this general framework, \u003cem\u003eAnchomanes difformis\u003c/em\u003e (Blume) Engl. (family Araceae) appears to be a plant of interest. \u003cem\u003eAnchomanes difformis\u003c/em\u003e, locally called \"Adodo\" in Togo, is a medicinal plant used in traditional African medicine, particularly in Nigeria, Togo and C\u0026ocirc;te d'Ivoire [12,13]. It grows mainly in humid tropical areas, savannahs and along waterways at low or medium altitudes [12,13]. This species is a perennial herbaceous plant, with a horizontal tuber measuring between 40\u0026ndash;80 cm long and 8\u0026ndash;22 cm in diameter, with annular scars [12,13]. Each season, it produces a single large tripartite leaf, up to 1.5 m wide, with sessile leaflets of variable shape [12,13]. The petiole, greenish purple, reaches 2.5 m in length and bears small green needles [12,13]. The stem is green with a white base [12,13]. The inflorescence emerges from the tuber, with a cylindrical spadix bearing female flowers with purple-pink ovary at the base, followed by creamy-white male flowers [12,13]. It is widely used in traditional local medicine: roots, rhizomes and leaves are used against an impressive variety of ailments (abdominal pain, respiratory disorders, joint problems, infections, digestive disorders, neurological disorders, etc.) [12,13]. The roots of \u003cem\u003eA. difformis\u003c/em\u003e are used in Benin to treat anal wounds, oral wounds, dysentery, diabetes and its complications and dysentery [14]. Other surveys have shown that all parts of the plant treat asthma [15\u0026ndash;17], malaria [18], cough and throat-related problems [13]. Some investigations have reported the use of powdered root mixed with palm oil is used as a remedy for respiratory diseases in children in Zaire [19]. The antimicrobial properties of the plant have been reported [16,20]. It is cited as a diuretic and \"all-comer\" remedy in the Togolese and Nigerian pharmacopoeia [12,13].\u003c/p\u003e\u003cp\u003eHowever, despite its widespread traditional use, the available scientific data on this species remain limited. As noted in recent work, very few studies have characterized the chemical composition or assessed the biological activities of \u003cem\u003eA. difformis\u003c/em\u003e [21\u0026ndash;26]. The lack of valid data makes a rigorous review of its active ingredients even more relevant.\u003c/p\u003e\u003cp\u003eIn this perspective, the present study aims to fill these gaps with a systematic approach: it aims to analyze the phytochemical composition of the hydroethanolic extract of \u003cem\u003eAnchomanes difformis\u003c/em\u003e and to evaluate its potential \u003cem\u003ein vitro\u003c/em\u003e biological activities (in antioxidant and antimicrobial). The aim is to provide a scientific basis for the traditional uses of this plant and to search for new therapeutic agents of plant origin. This approach is in line with the current need to discover, within plant biodiversity, molecules capable of effectively fighting antibiotic-resistant pathogens [4,6].\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003eCollection of plant material\u003c/h2\u003e\n \u003cp\u003eLeaves (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e), rhizomes and bulbs (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e) of \u003cem\u003eAnchomanes difformis\u003c/em\u003e were collected in Dalav\u0026eacute; (Ts\u0026eacute;vi\u0026eacute;, Maritime Region of Togo) in October 2021. The material plants have been botanically authenticated at the Herbarium of the Department of Botany, Faculty of Sciences, University of Lome.\u003c/p\u003e\n\u003c/div\u003e\n\u003ch3\u003eBacterial strains\u003c/h3\u003e\n\u003cp\u003eThe microorganisms used for the test were clinical and reference strains of \u003cem\u003eStreptococcus pneumoniae\u003c/em\u003e, \u003cem\u003eEscherichia coli\u003c/em\u003e, \u003cem\u003eStaphylococcus aureus\u003c/em\u003e and \u003cem\u003eCandida albicans\u003c/em\u003e, all isolated at the National Institute of Hygiene in Lom\u0026eacute; (INH-Lom\u0026eacute;).\u003c/p\u003e\n\u003ch3\u003eExtraction\u003c/h3\u003e\n\u003cp\u003eThe extraction was carried out in accordance with[27] and taken up by [28]. The samples of each plant were carefully washed under running water and dried in the open air at room temperature for two weeks. After drying, the plant materials were reduced to powder using the grinding machine. The ethanolic extraction was carried out by macerating 100 g of powder in 1,000 ml of 70% ethanol (v/v), under continuous stirring for 48 hours at room temperature. The extract was filtered through WHATMAN N\u0026deg;1 filter paper and evaporated at 45\u0026deg;C using a rotary evaporator until dryness under reduced pressure. The extracts were stored at 4\u0026deg;C in the refrigerator until used.\u003c/p\u003e\n\u003cp\u003e\u003cimg 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bIwxLaIjygsWLEi7d++ut4wxxowDdBf9FR1R5hP3mrXKGGPMeEB30V/REWWmSdy5c2e9ZYwxZhygu5qmFjqifM0116QtW7bUW8YYY8YBurt8+fJ6q2gpL126NK1bt672McYYM0rQW3R32bJltU+Y+wKYcHzJkiVpw4YNefJuY4wxo4GPLqxevTrt2LEjTU1N1b6hpQxMh7h+/fq0YsWKHMEYY8zwQV/RWfQ2CjJ0iTKsWrUqf/4FBbcpwxhjhgu6ir6is+htSZf5IsIHNO+55540PT2dVq5cmRYvXpw/nLhw4cI6hDHGmEEwDplhb4yy4E89bMi0kLFMNNFTlMX27dvzF3GZNJyPTe7Zs6feY4wxZhC8GMI4ZAZTMMotDn9rYqAoG2OMGRcp/RfCWNGl9+q7ZwAAAABJRU5ErkJggg==\"\u003e\u003c/p\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n \u003ch2\u003ePhytochemical analysis\u003c/h2\u003e\n \u003cp\u003ePhytochemical screening was performed based on staining characteristic tests to highlight major chemical groups. It focused on the hydroethanolic extracts of the plants studied. The chemical groups were identified by reference to the methods described by [29] and taken up by [30\u0026ndash;32].\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\n \u003ch2\u003eAlkaloids\u003c/h2\u003e\n \u003cp\u003eThe dissolved extracts were divided into two tubes at a rate of 2 mL each. In a first tube (test tube), 4 mL of sulphuric acid (H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e) diluted to 10% and a few drops of Dragendorff\u0026apos;s reagent were added. The appearance of an orange-red precipitate indicates the presence of alkaloids in the hydroethanolic extract. The second tube served as a control.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003ch3\u003eFlavonoids\u003c/h3\u003e\n\u003cp\u003eThe dissolved extracts were divided into two tubes at a rate of 2 mL each. In a first tube (test tube), 4 mL of methanol and a few drops of magnesium turns were added; then 1 mL of concentrated HCl was added. The appearance of a red coloration indicates the presence of flavonoids in the hydroethanolic extract. The second tube served as a control.\u003c/p\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003eSaponines\u003c/h2\u003e\n \u003cp\u003eThe dissolved extracts were divided into two tubes at a rate of 2 mL each. In a first tube (test tube), a few drops of distilled water were added. The appearance and persistence of foam after stirring indicates the presence of saponosides. The second tube served as a control.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003eTannins\u003c/h2\u003e\n \u003cp\u003eThe dissolved extracts were divided into two tubes at a rate of 4 mL each. In a first tube (test tube), a few drops of FeCl\u003csub\u003e3\u003c/sub\u003e were added. The appearance of a blackish brown colour after homogenization of the mixture indicates the presence of tannins in the hydroethanolic extract. The second tube served as a control.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003ePhenols\u003c/h2\u003e\n \u003cp\u003eThe dissolved extracts were divided into two tubes at a rate of 4 mL each. In a first tube (test tube), a few drops of FeCl\u003csub\u003e3\u003c/sub\u003e were added. The appearance of a blackish brown colour after homogenization of the mixture indicates the presence of phenolic compounds in the hydroethanolic extract.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003eTriterpenes and sterols\u003c/h2\u003e\n \u003cp\u003eThe dissolved extracts were divided into two tubes at a rate of 4 mL each. In a first tube (test tube), 1.6 mL of chloroform and 2.6 mL of sulphuric acid (H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e) were added. The appearance of a reddish-brown ring between two phases, one clear at the bottom and the other green at the top, indicates the presence of triterpenes and sterols in the hydroethanolic extract. The second tube served as a control.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n \u003ch2\u003eCoumarins\u003c/h2\u003e\n \u003cp\u003eThe dissolved extracts were divided into two tubes at a rate of 4 mL each. In a first tube (test tube), 2 mL of distilled water was added, and a small amount of ammonia was added to the mixture. The appearance of fluorescence after ultraviolet illumination indicates the presence of coumarins in the hydroethanolic extract. The second tube served as a control.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003eReducing sugars\u003c/h2\u003e\n \u003cp\u003eThe dissolved extracts were divided into two tubes at a rate of 2 mL each. In a first tube (test tube), 2 mL of Fehling\u0026apos;s Liquor (1 mL of solution A\u0026thinsp;+\u0026thinsp;1 mL of solution B) were added and brought to a boil in a boiling sea bath for 2 to 3 min. The appearance of a brick-red precipitate indicates the presence of reducing sugars in the hydroethanolic extract. The second tube served as a control.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003eFlavonoid content\u003c/h2\u003e\n \u003cp\u003eThe flavonoid content is estimated according to the method described by [33] and taken up by [30\u0026ndash;32]. It consisted of preparing plant extracts at 1mg/mL in distilled water, aluminum chloride (AlCl\u003csub\u003e3\u003c/sub\u003e) 2% in distilled water, and rutin at different concentrations of 0; 5 ; 25 ; 50 ; 75 ; 100 ; 150 ; 200 \u0026micro;g/mL in methanol. The operation consisted of mixing 1 mL of the extract solution at 1 mg/ml or 1 mL of each rutin concentration with 1 mL of aluminum chloride (AlCl\u003csub\u003e3\u003c/sub\u003e) 2% using a vortex. After 10 minutes of incubation, the absorbance was directly measured with a UV-visible spectrophotometer (METASH UV-5200PC UV/VIS Spectrophotometer) at 415 nm against a blank. Rutin was used as a standard. The total flavonoid content of the extracts was inferred from the calibration curve established with Rutin (0\u0026ndash;200 \u0026micro;g/mL) and the results were expressed in microgram equivalent of rutin per milligram of dry matter (\u0026micro;g ER/mg DM). Three tests were conducted for each extract. The total flavonoid content (\u003cstrong\u003eXFlav\u003c/strong\u003e) is obtained using the following formula:\u003c/p\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,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\"\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eXFlav\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eTotal flavonoid content (\u0026micro;g ER/mg DM)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eOD\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eOptical Density read at 415 nm\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n \u003ch2\u003eAntioxidant activity\u003c/h2\u003e\n \u003cdiv id=\"Sec19\" class=\"Section3\"\u003e\n \u003ch2\u003ePhosphomobyldate reduction test\u003c/h2\u003e\n \u003cp\u003eDetermination of antioxidant activity by the phosphomobyldate reduction assay was performed using the method described by [34] and slightly modified in [31,32]. The phosphomolybdate reagent was prepared (100 mL of reagent) from a mixture of 90 mL of 0.6 M sulphuric acid, 5 mL of 0.1% sodium phosphate and 5 mL of 1% ammonium molybdate. For the test, 1 mL of each extract was added to 9 mL of the above reagent. The whole thing was brought to a temperature of 95\u0026deg;C in a water bath for 90 minutes. Then the mixture was cooled to room temperature. Absorbances were measured at wavelengths of 820 nm against a blank consisting of reagent and distilled water. Ascorbic acid was used as the standard antioxidant under the same operating conditions and the results were expressed in milligrams of ascorbic acid equivalent per gram of dry matter (mg AAE/g DM). Three tests were performed for each concentration of product tested. The antioxidant activity related to the reducing power of the extracts is expressed in Antioxidant Content (X\u003csub\u003eMob\u003c/sub\u003e) using the following formula:\u003c/p\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAVMAAAA6CAYAAAANx7pSAAAAAXNSR0IArs4c6QAAAARnQU1BAACxjwv8YQUAAAAJcEhZcwAADsMAAA7DAcdvqGQAAAy+SURBVHhe7Z1JaBRPG8bL75z4x+USlSDGiyAoGlf0oOAehKi4XMSgqBEPBsVdzEHUiMQgxF0UDy4oUSRxBT0oKkZFMeBFRUTMJa7x7jdPWe/MO5XuWZKZZCZ5flB0dXVNT03XW09XV73V0+9vBEMIIaRL/M9tCSGEdAGKKSGEZACKKSGEZACKKSGEZICEE1CNjY2mubnZvHz50rx69cq0tra6I4QQ0ncoKioy48ePN+PGjTMTJkwwZWVl7kiMQDFtb283VVVV5unTp2bRokX2BDhRcXGxy0EIIX2Hz58/RzuVDQ0NZurUqaa2ttYUFha6HAFi2tTUZDZs2GDKy8vNkSNHTL9+/dwRQgghkEx0Nm/cuGHq6+vNggULbHqcmKJHOnr0aFNRUWGqq6tdKiGEEB9o5Pnz583bt29tDzVOTNesWWMKCgpMXV2dSyGEEBIGeqh//vwxp0+fjokpJpu2b99uVZaP9oQQkhzIJ57ma2pqYq5RmLXHOCmFlBBCUgN6iUl66GdUTDFLhVl7QgghqQPdxEx/9DF/yJAh5tmzZ3R/IoSQNIDb1OTJk2Niiu6qixJCCEkD6CeXk5KscvLkSTNw4EDz+PFjl0JI74RiSrLCx48fzdy5c8369evNjx8/XCohvReKKckKhw8ftu4ipaWlLoWQ3g3FlGSFY8eOmTFjxphBgwa5FEJ6NxRTQgjJABTTDIHZPAm5AMYrdZkQkOZz+fLl6HFMFGGsk5BscOvWraS2Jsd1OHTokDua2+SNmOJNVvoC+xWhj6Oivn37ZitBf2bXrl0udwzk03kQ+gq4hrhuYMCAAebhw4dmxIgRdp9kH7n+sFexPbwrEze4VIGXxPLly+PsF/u+94TfFoKC/oy0n5EjR0aPJyob0nFc8uJz+DzOI8yfP9+cOHHCxjEpKbbXa4CfKVDRnKStre1vpMHbciJUVla6I3//fvjwIZqO0NTUZNNramri0vF5nEfj50HoDF39fKaZM2dO0vLgGkqeiJG71MyBeikpKbHnv3TpkkvtOq9fv46zBR1KS0vt78J3+6DucUx/FuULy59N8H1hvwFh586dLmc4sPOgz0qQdgCC7NwPjx49snlxnXAdg/Ig6LYHtB35Aefx25y2zTC7QFkkD8qe69iyurjdyXV845EGoCtz2bJlNg0EGZCuGF+gJXSGrn4+0yQTU1w7OQ5ByTRhjTdT+I0NdYmGKeKNetViosVLGjjOIWnYpiuo8htxrdPFt1kpj6QhJCuP/FYEsWt93XW96vRk5dVlk2uFG5ikIYjw4ppLGq4h8iF/UNmEVGyPYtoNaJFApeuLjspERQragCToytOGoIMG54exS6NDwL5uqEB/HgaFPDq/Lld3kExM0fOR45nsNXYXYY0N11kLpFx3EQjfRvR5kCcdxL46I6bynQgog6DrLZGI6HIjaHS6nFu3hWTl1bau7VyLrFwrbee6N63bVpBg6nOhvfiE1W+ugnLm3QQUXG4ilW3jx48fN6tWrbJxgGPJXHEid8XouM/u3bvtNlLZduuDfNOnTzdXrlyJczzHPt6ujdU9QcyYMcPmERAPmvzxCZo0Cgtd5d69ey5mzKxZs1ws/0H941WSAHV2//59OzYJWwEYT9Q2Mm3atGj9S55s449njho1ysWM/Xsg4cGDBy7WkSdPnriY6eDLq/d1PuHu3btRO8I4pz+2qW29f//+LmbM8OHDXezfeC/4+fOn3YL//vvPxYwZNmyYi/1rcz76P5RQnt5A3okpJkgOHDjg9mIVFbnb2oYSBo5Lo4GIQijls2vXrrVbDYxLD5BH7rS4/ZvIHdOlGLu6x58IAzdv3uyQ98WLF3Y2M1dAeQCuS2/zBcX/8wh4CcXXr1/dXrwgCFp83rx542Ldh77+WpBSxa+/dOoTdrBt27bQm/3v379dLDm/fv1yseRMmjTJxRLfNPKJvHSNWrJkiYvF0D3UMEQ0IaLSK0VD0o1PQI9G7tDII0KNnkzk0cbGwbVr11wsBvLIVudtaWlxsWDu3LljRTiV0BV0z0j3hHojaKhBvTONfvUk/ronDP1kgAARArqnJ8HvffY0W7dujbOftrY2u0JNgKjKk5Z+Utu7d2+01/rp0ye71WjvD/Ts5Wb05csXuw0Dgi9PmO/fv7fbfCcvxfTs2bMuFgPiqB9Vgli9erWLxXq0mzdvtlsf9GgE/06vG1+yu2quvyM2qCfkC0OuhXSgq1cwsGkIrO6VwzUOHD161G4BRHbw4MH2ugcNg2zZsiUqiuh8jB071uZdsWKFTUvExIkT7TZoGCAfyTsxxZ1PegSoRDymAlQI/no1ETCgyspKt/fvDpxoaCATpPPo051jponQPZhcDOmAx3r8rUQidI8L7/UNwy+H9Oxgg/4xeTpJBd0JSMdeBL8TkaxTodEdBRn/hD9oU1NTnNCi3ei2I080uFlBhPUTGK6H7vXq8/Rm8k5M8ad/AsZO5XEd7N+/P+nj1bp161wseKxU0C/J9o1TN76ZM2e6WDB6oqcz42HZpjONN9fRj/UQ0qFDh7q9f/8o4YO/nAC4uXZHT9YX2nfv3rmYsW9sFxLZlh6akvFvQe9LPkw0BYmsTtPfB0HFdZGbAyZ3tS0vXbrUxYx9BwPmICQvhqt0G9FtVvP8+XO71cMKeU3kx1tUNGfR7h2RCnCp8e4kkbugS03dHSTIzUS72CCI+5CfN9Ijtuk6TVxv4Aiv0yVvd6CvSRByLNF1yWXCXGd0vWlbgL1Ifl0PYedJBbGvzlxD7RokrnNhtqXtWJdR/yZJ13l1G5F9cXXC9+m8CGH2GeTqFwbO4ZdB2oMGaZIn6Pp1pV56AltWF7c7uYzvNKx9I1GB+liQYaUrpkD7ygUFvWoo6LgO3W0QycQUQiPHg4w91/EbG34DhEJ+FwRV+y/q/GILSJP82KZ7HcS+OiOmsFl9s/aD9tnUdqztCL9Xf8YP2kc06LgOuj2BoDwI+K3+ddK2pgOENMiHFOiyB7UNimkW0Y0/yHj1nR7Bv0MmMvgwMQU4hjuxNnzsa0MFcgz5tHHBoHxD7Q50GYLIZ6d9NNAwIYKd4LcFCaPfw5L8sJOeuKHARmG3+regPH59aDv2hUXsU44jYB/pGtgrvgv2KPnwvUF5gS6T5AuzE//7YXvoaCS6prq9BgmubpMUU9KjJBNT3ZtHA/OBgcs50JjC1u6jweiG4QsZGrE0YGx1w9UiIUEaji6/Dvkm/KQjyWwP5KOY5qVrFEkP8QDQjtmYaIn0KGw8YtwdVnMtXrzY+v9FhNHMnj3bLlAImtzbs2ePdZmJiK99IxAmAcV1DQsasFIs0tuy5wELFy5Ma7ZZExH1rHtfkOwTueG6mDH79u1zsX+IrWLlYb5BMe3DQPggUGDHjh1Rh2uIJgQWYgvXmY0bN9r0ixcv2q0GQgqxxIyueEqcOnXKbmVBw8qVK+154D0hSzyFSK8DXedogO+jAAGWdOSTZaIkf8FNW5ZaR54+etXNkWLaSwlaTYU0DXqncHkBEDm8UwC9SXEt8l25/KWz0lPVvooAQgxkQYNe3w30gogwUFY5L3qyBw8eDFz5RvIHLKfGEw7ATVxsT+PbLIK+weYyFNM+DnoGYrTfv3/v9hVD6MXqYYigtfEYNsBQQ3eXjWQW+K72pK1lG4op6TGwvBeO4GhcGG/FGndZ3SagN4w0DBUQkstQTEkHZPmlvzrKfymKvDrOn1DCGCqQ/P6bh+T8eIyX3omMt/qvY8NfRpeUlNheDSG5DMWUdACvR8OYFiYKIJS3b9+26fKiGDyOY7wUYohJBCxfxOO5vGJQXh4zb948u71w4YLdXr16NU4Y8Wgv/8slnxUPA4BeKSa4Ei37JSRniDxiWVSUEOtnqv1DtX8n9sXRGj6l2h9Vr9wB+BzScRwO6dpBG87ecgxb+KtqH1XxQ4VfIiG5DOy0n4vYXoKLEhIKeqR4VyX9PQmJYSdRKaaEENI1oJ8cMyWEkAxAMSWEkAxAMSWEkAwQFdOioqKUlvkRQgiJAd2EfkbFFH/8pv8ygRBCSHKgm1igEhVT7AT9Pw4hhJBwoJvojEbFFH+41dDQQPcoQghJEejl9evXrX5G/UwB/kWwoKDA1NXVuRRCCCFhVFVVmfb2dnPmzJl4MUUiXkJRUVFhqqurXSohhBAfaOS5c+dMS0uLKSwsjHeNQgJe2IoMmzZt4iM/IYR4QBehj9DJ+vp6q5sgrmcqoIeK7uuzZ89MeXm5HVzFBFVxcbHLQQghfQe4P2HWHpNNGCOdMmWKqa2tjQopCBRTobGx0TQ3N9sT4EStra3uCCGE9B3gR4oOJTqWmGwqKytzR2IkFFNCCCGpYMz/AVttatRrvV5oAAAAAElFTkSuQmCC\"\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eXMob\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eAntioxidant Content (mg AAE/g DM)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eOD\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eOptical Density read at 820 nm\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\n \u003ch2\u003eTest FRAP (Ferric Reducing antioxidant Power)\u003c/h2\u003e\n \u003cp\u003eThe determination of antioxidant activity by the FRAP assay was carried out using the method described by [35] and taken up by [30\u0026ndash;32,36]. Thus, 3 mL of the freshly prepared FRAP test reagent in a test tube, 100 \u0026micro;L of the various solutions of iron II sulphate with concentrations between 0 and 2000 \u0026micro;mol. L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e will be added. The mixture was vigorously shaken at the vortex and the optical density was read after 5 min with a spectrophotometer at 593 nm. The absorbance of the TPTZ-Fe\u003csup\u003e2+\u003c/sup\u003e complex allowed to draw a calibration curve from the concentration range (0\u0026ndash;2000 \u0026micro;M) of the iron sulfate solution (FeSO\u003csub\u003e4\u003c/sub\u003e, 7H\u003csub\u003e2\u003c/sub\u003eO) dissolved in methanol. For the samples of the extracts to be tested, the FRAP reagent (3 mL) and the solution of the extract to be tested (100 \u0026micro;L) with a strength of 1 mg/mL were mixed in the same proportions as for the standard curve plotting. The optical density was read after 5 min at 593 nm. The antioxidant capacity of the extracts will be measured using the calibration curve by the color change related to the formation of the complex (Fe\u003csup\u003e2+\u003c/sup\u003e TPTZ) and expressed in micromole equivalent of iron sulfate per milligram of dry matter (\u0026micro;mol Eq FeS0\u003csub\u003e4\u003c/sub\u003e/mg DM). The tests will be repeated 3 times. The antioxidant activity related to the reducing power of the extracts is expressed in Reducing Power (X\u003csub\u003eFRAP)\u003c/sub\u003e using the following formula:\u003c/p\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,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\"\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eXFRAP\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eReducing power (\u0026micro;mol Eq FeS0\u003csub\u003e4\u003c/sub\u003e/mg DM)\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eOD\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eoptical density read at 593 nm\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\n \u003ch2\u003eEvaluation of the antimicrobial potency of extracts\u003c/h2\u003e\n \u003cp\u003eThe identification of the active extracts was carried out by the solid-state diffusion method [37] and the determination of the minimum inhibitory concentration (MIC) by the liquid dilution method [38].\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e\n \u003ch2\u003ePreparation of microbial strains\u003c/h2\u003e\n \u003cp\u003eAfter collection, the strains were transplanted to their appropriate media (Chapman for \u003cem\u003eS. aureus\u003c/em\u003e, Sabouraud\u0026thinsp;+\u0026thinsp;Chloramphenicol for \u003cem\u003eC. albicans\u003c/em\u003e, Mac Conkey for \u003cem\u003eE. coli\u003c/em\u003e and Fresh Blood Agar (FSG) for \u003cem\u003eS. pneumoniae\u003c/em\u003e) and stored. To obtain the 24-hour colonies that were used to prepare the inoculum, the strains were transplanted on agar medium without inhibitors [39].\u003c/p\u003e\n \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e\n \u003ch2\u003ePreparation of the inoculum\u003c/h2\u003e\n \u003cp\u003eFrom an 18\u0026ndash;24 h culture on agar medium, a suspension was prepared in saline solution (0.9% NaCl) equivalent to the Mac Farland standard 0.5 (~\u0026thinsp;10\u003csup\u003e8\u003c/sup\u003e CFU/ml). Inoculation was sown by flooding with the inoculum suspension diluted to 1/100 (~\u0026thinsp;10\u003csup\u003e6\u003c/sup\u003e CFU/ml) in accordance with the necessary safety measures [39].\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec24\" class=\"Section2\"\u003e\n \u003ch2\u003ePreparation and sterilization of the stock solution of extracts\u003c/h2\u003e\n \u003cp\u003eThe extracts were dissolved in sterile distilled water to an initial concentration of 200 mg/mL (200 mg of each extract in 1 mL of distilled water). To verify their sterility, an aliquot of each stock solution was taken and inoculated on Muller Hinton agar. After incubation at 37\u003csup\u003e\u0026deg;C\u003c/sup\u003e for 24 hours, those that gave colonies were filtered with a millipore membrane of 0.45 \u0026micro;m in diameter. After sterilization, each stock solution is diluted 1/4 with distilled water to obtain solutions with a concentration of 50 mg/mL of each extract.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eIn vitro\u003c/strong\u003e \u003cstrong\u003eantimicrobial activity\u003c/strong\u003e\u003c/p\u003e\n \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e\n \u003ch2\u003eAntimicrobial testing of extracts on germs tested by the well method\u003c/h2\u003e\n \u003cp\u003eMueller Hinton (MH) agar plates for bacteria and Sabouraud dextrose\u0026thinsp;+\u0026thinsp;Chloramphenicol for yeasts were inoculated by flooding following the recommendations of the Antibiogram Committee of the French Society of Microbiology [39]. After the dishes have dried, (about 5 min) the agar is perforated into a maximum of six wells (90 mm Petri dishes) with a sterile tip previously cut so that the diameter is about 6 mm. The cavities formed are filled with 50 \u0026micro;L of the 50 mg/mL extract solution (50 \u0026micro;L per well). The boxes are placed at room temperature 25\u0026deg;C for 15 minutes for a pre-diffusion phase. Incubated at 37\u003csup\u003e\u0026deg;C\u003c/sup\u003e for 24 hours for bacteria and 48 hours for yeasts. Ciprofloxacin (bacteria), Nystatin (yeast) were used as a positive control and distilled water as a negative control. After 24 hours of incubation, the diameters of the inhibition zones were measured using a graduated ruler. The tests are carried out three times and an average are obtained [37]. MICs and BMCs were determined for extracts with an inhibition diameter greater than or equal to 11 mm.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e\n \u003ch2\u003eAntimicrobial testing of extracts on germs tested by the disc method\u003c/h2\u003e\n \u003cp\u003eMueller Hinton (MH) agar plates for bacteria and Sabouraud dextrose\u0026thinsp;+\u0026thinsp;Chloramphenicol for yeasts is dried (about 5 min) and inoculated by flooding according to the recommendations of the Antibiogram Committee of the French Society of Microbiology [39].\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eDisc preparation\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eWATTMAN (No. 1) 6 mm diameter, virgin, autoclaved paper was impregnated with 50 \u0026micro;L of the extracts at the initial concentration of 50 mg/mL. The final load of the disc is 2.5 mg. WATTMAN N\u0026deg;1 papers are dried in an oven (37\u003csup\u003e\u0026deg;C\u003c/sup\u003e).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eSensitivity test\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThese discs are placed aseptically on the previously inoculated agar. The boxes are placed at room temperature 25\u003csup\u003e\u0026deg;C\u003c/sup\u003e for 15 minutes for a pre-diffusion phase. Incubated at 37\u003csup\u003e\u0026deg;C\u003c/sup\u003e for 24 hours for bacteria and 48 hours for yeasts. Ciprofloxacin (bacteria), Nystatin (yeast) were used as a positive control and distilled water as a negative control. After 24 hours of incubation, the inhibition diameters were measured. The tests are carried out three times and an average are obtained on the three determinations [37]. MICs and BMCs were determined for extracts with an inhibition diameter greater than or equal to 11 mm.\u003c/p\u003e\n \u003cp\u003eDetermination of the minimum inhibitory concentration (MIC)\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec27\" class=\"Section3\"\u003e\n \u003cp\u003eThe minimum inhibitory concentration (MIC) was determined by the microplate microdilution method [38]. Mueller Hinton Broth is used to prepare serial dilutions at half concentrations ranging from 50 to 0.097 mg/mL. The inoculum (10 \u0026micro;L) was added to each cup containing 50 \u0026micro;L of extract. Cupules without inoculum were considered as negative controls. All these cups were incubated at 37\u0026deg;C for 24 h. The first cup in the series showing no sign of culture visible to the naked eye was MIC. An aliquot of the contents of each cupule with no visible growth was taken and then seeded by spreading on Mueller Hinton medium [38]. The dishes are incubated at 37\u0026deg;C for 24 to 48 hours at the end of which the number of colonies is determined. After incubation, the smallest concentration of the extract that does not produce colonies is considered the MBC. To determine whether the antimicrobial effect observed is bactericidal or bacteriostatic, the MBC/CMI ratio is used to assess the activity of the extracts (bactericidal/bacteriostatic).\u003c/p\u003e\n \u003cul\u003e\n \u003cli\u003e\n \u003cp\u003eFor a CMB/MIC ratio greater than 1, the antimicrobial effect is bacteriostatic.[40]\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eFor a CMB/MIC ratio of 1, the antimicrobial effect is bactericidal [40].\u003c/p\u003e\n \u003c/li\u003e\n \u003c/ul\u003eStatistical analysis of the data\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec28\" class=\"Section2\"\u003e\n \u003cp\u003eThe data obtained was subjected to statistical analysis using RStidio 4.1.3 software from 10/03/2022 and Microsoft\u0026reg; Excel\u0026reg; spreadsheet for Microsoft 365 MSO 2021. Quantitative variables (flavonoid content, FRAP test, and phosphomobyldate reduction method) are presented as a mean and standard deviation. One-Factor Analysis of Variance (ANOVA) was used to assess the diameters of the zones of inhibition. The significance level has been set at 5%.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eFor this study, the best yield is obtained by the extract of the leaves of \u003cem\u003eA. difformis\u003c/em\u003e (9.47%). The weakest is obtained with the extract of the rhizomes of \u003cem\u003eA. difformis\u003c/em\u003e (4.10%). All extracts have an acidic pH (pH ˂ 7) and range from 6.28 to 5.40. The solubility is quite good, compared to the solvent used for all the extracts. The colour of the extracts is light yellow with a sticky gum appearance for all three extracts (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe extracts were analyzed for alkaloids, tannins, flavonoids, saponosides, reducing sugars, triterpenoids, and sterols. The result is recorded in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Hydroethanolic extracts of leaves, rhizomes and bulbs \u003cem\u003eof A. difformis\u003c/em\u003e contain only alkaloids, coumarins and reducing sugars (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eQuantitative tests show a low presence of flavonoids in all extracts at various doses (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The highest flavonoid content was measured with the hydroethanolic extract of the bulb of \u003cem\u003eA. difformis\u003c/em\u003e (33.78 \u0026micro;g ER/mg DM) and the lowest with the hydroethanolic extract of the leaves of \u003cem\u003eA. difformis\u003c/em\u003e (28.85 \u0026micro;g ER/mg DM).\u003c/p\u003e\u003cp\u003eThe evaluation of antioxidant activity revealed that all extracts have antioxidant power. The best antioxidant activity was observed with \u003cem\u003eA. difformis\u003c/em\u003e leaf extract with 37.79 mg AAE/g DM by the phosphomolybdate reduction assay and 270.50 \u0026micro;mol FeS0\u003csub\u003e4\u003c/sub\u003e/mg DM by the FRAP assay (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eRegarding antimicrobial activity, at the initial concentration of 50 mg/ml, the three hydroethanolic extracts tested in this work did not significantly inhibit the growth of microorganisms (p\u0026thinsp;=\u0026thinsp;2.e-16, cv\u0026thinsp;=\u0026thinsp;3.862\u0026ndash;5.562%) with the two diffusion methods on all strains used (Tables\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e \u0026amp; \u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). Therefore, the MIC was not determined (Inhibition Diameter ˂11 mm).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eSome characteristics of the extracts obtained\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePlants\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOrgans\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAspect\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eColour\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eSolubility\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003epH\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eYield\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e\u003cp\u003e\u003cem\u003eA. difformis\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLeaves\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSg\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLy\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePg\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5,40\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e9,47%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBulbs\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSg\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLy\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePg\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e6,28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e4,14%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRhizomes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eSg\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eLy\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ePg\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e5,65\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e4,10%\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eSg: Sticky Gum; Ly: Light yellow; pH: Hydrogen potential; Pg: Pretty good\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePhytochemical study of the studied extracts\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eSecondary metabolites\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003eExtraits HE of \u003cem\u003eA. difformis\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLeaves\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBulbs\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eRhizomes\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAlkaloids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFlavonoids\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTannins\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCoumarins\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSaponosides\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTriterpenes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePhenol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSugar Reducers\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003eLegend: -: Absence; +: Presence; HE: Hydroethanolic\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eFlavonoid content of the extracts studied\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eHE Extracts\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003e\u003cem\u003eA. difformis\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLeaves\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBulbs\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eRhizomes\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTotal flavonoid content (\u0026micro;g ER/mg DM)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e24,85\u0026thinsp;\u0026plusmn;\u0026thinsp;0,26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e33,78\u0026thinsp;\u0026plusmn;\u0026thinsp;0,66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e29,82\u0026thinsp;\u0026plusmn;\u0026thinsp;0,88\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003eER: Equivalent to Rutine; HE: Hydroethanolic; DM: Dry matter\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAntioxidant activities \u003cem\u003ein vitro\u003c/em\u003e Excerpts studied\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHydroethanolic extracts\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePhosphomolybdate Reduction Test (mg AAE/g DM)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eFRAP Testing\u003c/p\u003e\u003cp\u003e(\u0026micro;mol FeS0\u003csub\u003e4\u003c/sub\u003e/mg DM)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFeuilles de \u003cem\u003eA. difformis\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e37,79\u0026thinsp;\u0026plusmn;\u0026thinsp;0,18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e270,50\u0026thinsp;\u0026plusmn;\u0026thinsp;5,00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBulbs of \u003cem\u003eA. difformis\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e19,59\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e202,17\u0026thinsp;\u0026plusmn;\u0026thinsp;2,89\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRhizomes of \u003cem\u003eA. difformis\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e23,09\u0026thinsp;\u0026plusmn;\u0026thinsp;0,20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e190,50\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"3\"\u003eAAE: Acide Ascorbique Equivalent ; FRAP: Ferric Reducing antioxidant Power ; DM: Dry matter\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\n\u003ch3\u003eAntimicrobial activities\u003c/h3\u003e\n\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAntimicrobial testing result of extracts on germs tested by The well method\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"8\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eMicrobial strains\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003eExtraits HE of \u003cem\u003eA. difformis\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eWater-Ethan\u003c/p\u003e\u003cp\u003eNC\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eCyprus/Nyst.\u003c/p\u003e\u003cp\u003ePC (50 \u0026micro;g/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eP-value\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eCV(%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLeaves\u003c/p\u003e\u003cp\u003e(50 mg/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBulbs\u003c/p\u003e\u003cp\u003e(50 mg/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eRhizomes\u003c/p\u003e\u003cp\u003e(50 mg/mL)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eS. aureus\u003c/em\u003e ATCC 29213\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e37.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3,856\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eS. aureus\u003c/em\u003e 0689\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e25.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3,833\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eE. coli\u003c/em\u003e ATCC 25922\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e44.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e4,225\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eE. coli\u003c/em\u003e 1628\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e17.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e4,037\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eS. pneumonia\u003c/em\u003e ATCC 49619\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3,922\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eS. pneumonia\u003c/em\u003e 034\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e33.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3,362\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eC. albicans\u003c/em\u003e ATCC 10231\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e32.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e4,495\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eC. albicans\u003c/em\u003e 1134\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5.562\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eThe values are expressed as the Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;Standard Error. Values in the same column followed by the same lowercase letter are statistically identical (Duncan, p ˂ 0.05)\u003c/p\u003e\u003cp\u003eHE: Hydroethanolic; NC: Negative control; PC: positive control; CV: Coefficient of Variance\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eResult of antimicrobial testing of extracts on germs tested by the disc method\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"8\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eMicrobial strains\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e\u003cp\u003eExtraits HE of \u003cem\u003eA. difformis\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eWater-Ethan\u003c/p\u003e\u003cp\u003eNC\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eCyprus/Nyst.\u003c/p\u003e\u003cp\u003ePC (50 \u0026micro;g/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eP-value\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eCV(%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eLeaves\u003c/p\u003e\u003cp\u003e(50 mg/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eBulbs\u003c/p\u003e\u003cp\u003e(50 mg/mL)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eRhizomes\u003c/p\u003e\u003cp\u003e(50 mg/mL)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eS. aureus\u003c/em\u003e ATCC 29213\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e38.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2,91\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eS. aureus\u003c/em\u003e 0689\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e25.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5,27\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eE. coli\u003c/em\u003e ATCC 25922\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e44.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5,95\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eE. coli\u003c/em\u003e 1628\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e18.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e1.8 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e6,92\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eS. pneumonia\u003c/em\u003e ATCC 49619\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5,05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eS. pneumonia\u003c/em\u003e 034\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e32.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e5,80\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eC. albicans\u003c/em\u003e ATCC 10231\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e32.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e11,81\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eC. albicans\u003c/em\u003e 1134\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6,00\u0026thinsp;\u0026plusmn;\u0026thinsp;0,00\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e31.00\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2 \u003csup\u003ee\u0026minus;16 ***\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e6.06\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eThe values are expressed as the Mean\u0026thinsp;\u0026plusmn;\u0026thinsp;Standard Error. Values in the same column followed by the same lowercase letter are statistically identical (Duncan, p ˂ 0.05)\u003c/p\u003e\u003cp\u003eHE: Hydroethanolic; NC: Negative control; PC: positive control; CV: Coefficient of Variance\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe study evaluated the antimicrobial activities of \u003cem\u003eA. difformis\u003c/em\u003e on clinical strains of \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eS. pneumoniae\u003c/em\u003e, and \u003cem\u003eC. albicans\u003c/em\u003e. The choice of this plant as study material is guided by their common use in traditional medicine in Africa to cure various diseases.[41]\u003c/p\u003e\u003cdiv id=\"Sec32\" class=\"Section2\"\u003e\u003ch2\u003e- Yield\u003c/h2\u003e\u003cp\u003eOur results show that for this study, the best yield is obtained by the extract of \u003cem\u003eA. difformis leaves\u003c/em\u003e (9.47%). The lowest is obtained with the extract of the rhizomes of \u003cem\u003eA. difformis\u003c/em\u003e (4.10%). These low yields could be explained by the short time (48 hours) and especially the mode and speed of rotation to homogenize the solvent and plant powder mixture[42]. Hydroethanolic extraction is valued because of its ability to extract a wide range of compounds, its relative safety of use, and its low cost compared to other extraction methods.[43]\u003c/p\u003e\u003cdiv id=\"Sec33\" class=\"Section3\"\u003e\u003ch2\u003e- Qualitative and quantitative phytochemical study\u003c/h2\u003e\u003cp\u003eThe qualitative phytochemical study of the three extracts of \u003cem\u003eA. difformis\u003c/em\u003e revealed the presence of alkaloids, coumarins and reducing sugars. These results do not corroborate those described by [15,44] who, in addition to the alkaloids obtained, also found tannins, flavonoids, saponosides and phenols but not reducing sugars. This could be explained by the extraction method used.\u003c/p\u003e\u003cp\u003eOur results showed a low presence of flavonoids in all extracts at various doses (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). This result confirms the absence of flavonoids in A. \u003cem\u003edifformis\u003c/em\u003e extracts in qualitative tests (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The lowest is that of the hydroethanolic extract of the leaves of \u003cem\u003eA. difformis\u003c/em\u003e (28.85 \u0026micro;g ER/mg DM). Our results corroborate with those of [17,21], who also found that the flavonoid content of \u003cem\u003eA. difformis\u003c/em\u003e leaves is lower with 4 mg ER/g DM using the same method.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec34\" class=\"Section3\"\u003e\u003ch2\u003e- Antioxidant activity\u003c/h2\u003e\u003cp\u003eRegarding antioxidant activity, the FRAP test and the phosphomobyldate reduction test gave a better reducing power of \u003cem\u003eA. difformis\u003c/em\u003e leaves with 270.5 \u0026micro;mol FeS0\u003csub\u003e4\u003c/sub\u003e/mg DM and 37.79 mg AAE/g DM respectively. The low presence of flavonoids and other secondary metabolites could explain this low antioxidant activity of the various extracts of \u003cem\u003eA. difformis\u003c/em\u003e [45].\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\n\u003ch3\u003e- Antimicrobial activity\u003c/h3\u003e\n\u003cp\u003ePresumptive testing by the agar diffusion method (Well and Disc) revealed that the three extracts did not significantly inhibit the growth of microorganisms (p\u0026thinsp;=\u0026thinsp;2.e\u003csup\u003e\u0026minus;\u0026thinsp;16\u003c/sup\u003e, cv\u0026thinsp;=\u0026thinsp;3.862\u0026ndash;5.562%) on all bacterial strains used for the study. Our results are in contradiction with the work of[12] which has shown that the essential oil extract of \u003cem\u003eA. difformis\u003c/em\u003e has antimicrobial activities on \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eC. albicans\u003c/em\u003e. This could be explained by the difference in the nature of the extracts used in the two studies. The low presence of phytochemicals is thought to be the basis for the antimicrobial inactivities observed with \u003cem\u003eA. difformis\u003c/em\u003e extracts [31].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe objective of our study was to evaluate the biological and phytochemical activities of \u003cem\u003eA. difformis\u003c/em\u003e on strains of \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eS. pneumoniae\u003c/em\u003e and \u003cem\u003eC. albicans\u003c/em\u003e. Qualitative phytochemicals have shown that the extracts (leaves, rhizome and bulb) of \u003cem\u003eA. difformis\u003c/em\u003e all contain coumarins, alkaloids and reducing sugars. Quantitative phytochemistry gave low total flavonoid content for all A. \u003cem\u003edifformis extracts in vitro\u003c/em\u003e. Our results showed that the three hydroethanolic extracts of \u003cem\u003eA. difformis\u003c/em\u003e did not significantly inhibit the growth of the microorganisms tested in this study.\u003c/p\u003e\u003cp\u003eUltimately, we can conclude that the hydroethanolic extracts of the leaves, rhizomes and bulbs of \u003cem\u003eAnchomanes difformis\u003c/em\u003e would be inactive on all the germs tested (\u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eS. pneumoniae\u003c/em\u003e and \u003cem\u003eC. albicans\u003c/em\u003e).\u003c/p\u003e\u003cp\u003eThis study sheds light on the chemical components and biological activities of the hydroethanolic extract of \u003cem\u003eAnchomanes difformis\u003c/em\u003e used in herbal medicine in Togo. Our results suggest that this plant could be a valuable source of bioactive compounds with potential applications in the treatment of various conditions other than the strain-related conditions tested in our study.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eATCC: American Typographic Culture Collection ; MIC: Minimum Inhibitory Concentration ; MBC: Minimum Bactericidal Concentration ; CS NDE: Sisters of Our Lady of the Church Health Center ; INH: National Institute of Hygiene of Lom\u0026eacute; ; MH: Mueller-Hinton ; WHO: World Health Organization\u003c/p\u003e\n"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics Approval and Consent to Participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot Applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot Applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical trial number\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of Data and Materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed in the current study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTchilabalo Bouyo, Sandrine T\u0026egrave;n\u0026egrave; Salifou, Jules Koffi Kpatagnon, Passimna Pissang and Komi Koukoura Komi harvested and identified the plant parts used. Tchilabalo Bouyo, Sandrine T\u0026egrave;n\u0026egrave; Salifou, Kodjovi Sossou, Samadou Tchakondo, Passimna Pissang, Yao Hoekou, Holaly Efui Gb\u0026eacute;kley and Komi Koukoura Komi participated in the extraction and flavonoid assay. Antioxidant activity tests were carried out by Tchilabalo Bouyo, Bawimodom Bidjada, Sandrine T\u0026egrave;n\u0026egrave; Salifou, Passimna Pissang, Jules Koffi Kpatagnon, Samadou Tchakondo, Abdoul Kader Ouedraogo, Isidore Kodjovi Anani Gbenonssi, Kodjovi Sossou, Komlan Tchalla, Yao Hoekou and Holaly Efui Gb\u0026eacute;kley. The results were analyzed statistically by Tchilabalo Bouyo, Holaly Efui Gb\u0026eacute;kley, Blaise Etienne M\u0026apos;BOUMBA and Passimna Pissang. Finally, Tchilabalo Bouyo, Richard Kouyassa Dessougmba, Passimna Pissang, Komi Koukoura Komi wrote the maniscrit and Komi Koukoura Komi and Tchadjobo Tchacondo approved and supervised the work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding declaration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eLack of funding for this study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003cbr\u003e\u003c/strong\u003eThe authors express their sincere thanks and deep gratitude to all those who have contributed to the realization of this work. Our thanks to the National Institute of Hygiene of Lom\u0026eacute; (INH-Lom\u0026eacute;) who provided us with the strains for the experiment and to the health center of the Sisters of Our Lady of the Church of Tokoin Seminary of Lom\u0026eacute; (CS NDE) for their moral support throughout the study process.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eLandecker H. 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Antioxidant Capacity, Phytochemical Analysis and Identification of Active Compounds in \u003cem\u003eAnchomanes difformis\u003c/em\u003e. Nat Prod J. 2020 Apr 22;10(4):446\u0026ndash;58. \u003c/li\u003e\n\u003cli\u003eOyetayo V. Comparative Studies of the Phytochemical and Antimicrobial Properties of the Leaf, Stem and Tuber of \u003cem\u003eAnchomanes difformis\u003c/em\u003e. Journal of Pharmacology and Toxicology. 2007;2(4):407\u0026ndash;10. \u003c/li\u003e\n\u003cli\u003eOghale O, Idu M. Phytochemistry, anti-asthmatic and antioxidant activities of \u003cem\u003eAnchomanes difformis\u003c/em\u003e (Blume) Engl. leaf extract. Asian Pac J Trop Biomed. 2016 Dec 31;6(3):225\u0026ndash;31. \u003c/li\u003e\n\u003cli\u003eOlanlokun J, Babarinde C, Olorunsogo O. Toxicity of \u003cem\u003eAnchomanes difformis\u003c/em\u003e, An Antimalarial Herb in Murine Models. European J Med Plants. 2017 Aug 28;20(3):1\u0026ndash;13. \u003c/li\u003e\n\u003cli\u003eDisengomoka I, Delaveau P, Sengele K. 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Available from: https://www.academia.edu/download/123085514/papers.pdf. \u003c/li\u003e\n\u003cli\u003eAsadi N, Bahmani M, Kheradmand A, Rafieian-Kopaei M. The impact of oxidative stress on testicular function and the role of antioxidants in improving it: A review. Vol. 11, Journal of Clinical and Diagnostic Research. 2017. \u003c/li\u003e\n\u003cli\u003eTchacondo T, Karou S, Batawila K, Agban A, Ouro-Bang K, Anani K, Gbeassor M, de Souza C. Herbal remedies and their adverse effects in Tem tribe traditional medicine in Togo. Afr J Tradit Complement Alter Med. 2011;8(1):45\u0026ndash;60. \u003c/li\u003e\n\u003cli\u003eOyenihi A, Opperman M, Alabi T, Mpahleni B, Masola B. Centella asiatica alleviates diabetes-induced changes in fatty acid profile and oxidative damage in rat testis. Andrologia. 2020 Jun 13;52(10):1\u0026ndash;12. \u003c/li\u003e\n\u003cli\u003eLawin IF, Laleye FOA, Agbani OP, Assogbadjo AE. Ethnobotanical assessment of the plant species used in the treatment of diabetes in the Sudano- Guinean zone of Benin. J Anim Plant Sci. 2015;26(3):4108\u0026ndash;23. \u003c/li\u003e\n\u003cli\u003eSita G. Traitement traditionnel de quelques maladies en pays Bissa (R\u0026eacute;publique de Haute-Volta). Burkina Faso; 1978 Jan. \u003c/li\u003e\n\u003cli\u003eKarou D, Savadogo A, Canini A, Yameogo S, Montesano C, Simpore J, Colizzi V, Traore AS. Antibacterial activity of alkaloids from \u003cem\u003eSida acuta\u003c/em\u003e. Afr J Biotechnol [Internet]. 2016 Jun 15 [cited 2022 Sep 30];5(2):195\u0026ndash;200. Available from: https://www.ajol.info/index.php/ajb/article/view/137755. \u003c/li\u003e\n\u003cli\u003eY. Hoekou, K. E. Poro, I. Kpabi, P. Pissang, Anoumou Y. Dagnra, K. M. Novidzro, Komlan Batawila, T. Tchacondo. In vitro Antimycobacterial Activity of Selected Medicinal Plants against \u003cem\u003eMycobacterium tuberculosis\u003c/em\u003e. Int J Curr Microbiol Appl Sci. 2021 Feb 20;10(2):3201\u0026ndash;8. \u003c/li\u003e\n\u003cli\u003eHarborne JB. Phytochemical methods: A guide to modern techniques of plant analysis. In: London, UK. 1998. \u003c/li\u003e\n\u003cli\u003eMiwonouko KF, Dermane A, Kombate B, Gbekley EH, Anani K, Metowogo K, Karou DS. Antioxidant and antimicrobial activities of Cassia alata (L.) Roxb. Leaves. Int J Biol Chem Sci [Internet]. 2024 May 9 [cited 2024 Jul 7];18(1):236\u0026ndash;43. Available from: https://www.ajol.info/index.php/ijbcs/article/view/269870 \u003c/li\u003e\n\u003cli\u003eTCHAKONDO S, BOLETI N, GAMAYIZOME AE, BOUYO T, GBEKLEY EH, TCHACONDO T, KAROU SD. Phytochemical, Toxicological, and Antimicrobial Evaluation of the Hydroethanolic Extract of Calotropis procera (Ait.) Leaves Used in Traditional Medicine in the Maritime Region of Togo: An Experimental Study. 2025 Jun 9 [cited 2025 Jun 30]; Available from: https://www.researchsquare.com \u003c/li\u003e\n\u003cli\u003eBouyo T, Komi K, Salifou S, Pissang P, Gbekley H. Phytochemical and biological studies of the hydroethanolic extract of \u003cem\u003ePteleopsis suberosa\u003c/em\u003e and \u003cem\u003ePiliostigma thonningii\u003c/em\u003e used in herbal medicine in Togo. 2025 [cited 2025 Aug 4]; Available from: https://www.researchgate.net/ \u003c/li\u003e\n\u003cli\u003eAndzi-Barh\u0026Atilde;\u0026copy; T, Massala K, Engonga L, Lebibi J. Phytochemical studies, total phenolic and flavonoids content and evaluation of antiradical activity of the extracts of the leaves from \u003cem\u003eDischistocalyx sp\u003c/em\u003e. (Acanthac\u0026Atilde;\u0026copy;es). J Pharmacogn Phytochem. 2015;3(6):174\u0026ndash;8. \u003c/li\u003e\n\u003cli\u003ePrieto P, Pineda M, Aguilar M. Spectrophotometric Quantitation of Antioxidant Capacity through the Formation of a \u003cem\u003ePhosphomolybdenum Complex\u003c/em\u003e: Specific Application to the Determination of Vitamin E. Anal Biochem. 1999 May 1;269(2):337\u0026ndash;41. \u003c/li\u003e\n\u003cli\u003eBenzie IFF, Strain JJ. The Ferric Reducing Ability of Plasma (FRAP) as a Measure of \u0026ldquo;Antioxidant Power\u0026rdquo;: The FRAP Assay. Anal Biochem. 1996 Jan 23;239(1):70\u0026ndash;6. \u003c/li\u003e\n\u003cli\u003eAgbodan K, Dotse K, Koumaglo H. Activit\u0026eacute;s antioxydantes des huiles essentielles de trois plantes aromatiques acclimat\u0026eacute;es au Togo. Int J Biol Chem Sci. 2014 Oct 17;8(3):1103\u0026ndash;10. \u003c/li\u003e\n\u003cli\u003eHegstad K, Giske CG, Haldorsen B, Matuschek E, Sch\u0026oslash;nning K, Leegaard TM, Kahlmeter G, Sundsfjord A. Performance of the EUCAST disk diffusion method, the CLSI agar screen method, and the Vitek 2 automated antimicrobial susceptibility testing system for detection of clinical isolates of enterococci with low- and medium-level VanB-type vancomycin resistance: A multicenter study. J Clin Microbiol. 2014 May 1;52(5):1582\u0026ndash;9. \u003c/li\u003e\n\u003cli\u003eGanfon H, Houvohessou JP, Assanhou AG, Bankole HS, Gbenou J. Activit\u0026eacute; antibact\u0026eacute;rienne de l\u0026rsquo;extrait \u0026eacute;thanolique et des fractions de \u003cem\u003eAnogeissus leiocarpa\u003c/em\u003e (DC) Guill. Et Perr. (Combretaceae). Int J Biol Chem Sci [Internet]. 2019 Aug 27 [cited 2022 Nov 27];13(2):643\u0026ndash;51. Available from: https://www.ajol.info/index.php/ijbcs/article/view/189305. \u003c/li\u003e\n\u003cli\u003eCA-SFM. Comit\u0026eacute; de l\u0026rsquo;antibiogramme de la Soci\u0026eacute;t\u0026eacute; Fran\u0026ccedil;aise de Microbiologie Recommandations 2022 V.1.0 Mai [Internet]. SFM/EUCAST, editor. Vol. 1.0. France; 2022. 1\u0026ndash;183 p. Available from: www.sfm-microbiologie.org. \u003c/li\u003e\n\u003cli\u003eOlajuyigbe O, Afolayan A. In vitro antibacterial activities of the methanol extract of Ziziphus mucronata Willd. subsp. mucronata Willd. Journal of Medicinal Plants Research . 2011 Aug 18;5(16):3791\u0026ndash;5. \u003c/li\u003e\n\u003cli\u003eOOAS. Pharmacop\u0026eacute;e de l\u0026rsquo;Afrique de l\u0026rsquo;ouest [Internet]. 2020 [cited 2022 Sep 25]. 1\u0026ndash;308 p. Available from: www.wahooas.org. \u003c/li\u003e\n\u003cli\u003ePatrick K, Kon\u0026eacute; FO, Soro Y, Siaka S. D\u0026eacute;termination des param\u0026egrave;tres influen\u0026ccedil;ant le rendement d\u0026rsquo;extraction hydro-alcoolique des m\u0026eacute;tabolites secondaires de Alchornea cordifolia (Euphorbiaceae) et Tridax procumbens linn (Asteraceae). J Soc Ouest-Afr Chim [Internet]. 2017 [cited 2025 Aug 4];16. Available from: http://www.soachim.org. \u003c/li\u003e\n\u003cli\u003eDai J, Mumper RJ. molecules Plant Phenolics: Extraction, Analysis and Their Antioxidant and Anticancer Properties. Molecules [Internet]. 2010;15:7313\u0026ndash;52. Available from: www.mdpi.com/journal/molecules. \u003c/li\u003e\n\u003cli\u003eQuartey AK, Korsah S, Apenteng JA, Nortey NND, Tagoe M, Mintah DN, Barfour AF, Owusu FWA, Kontoh DB. A Study of the Antimicrobial and Wound Healing Activities of the Ethanolic Leaf Extract of Anchomanes difformis (Blume) Engl. Pallidus. J Adv Med Pharm Sci. 2024 Apr 18;26(5):66\u0026ndash;75. \u003c/li\u003e\n\u003cli\u003eBenjamin SANOGO. \u0026Eacute;tude des activit\u0026eacute;s antioxydantes et anticonvulsivantes de deux (02) plantes m\u0026eacute;dicinales du Mali [Th\u0026egrave;se]. [Mali]: Universit\u0026eacute; de Bamako ; 2010. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Anchomanes difformis, phytochemistry, activity, antioxidant, antimicrobial","lastPublishedDoi":"10.21203/rs.3.rs-7401969/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7401969/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003eThe proliferation of strains that are multi-resistant to conventional antibiotics has led to a need for research into other active molecules. The aim of this study was to evaluate the antimicrobial activity of \u003cem\u003eAnchomanes difformis\u003c/em\u003e, a medicinal plant from the West African sub-region used in traditional pharmacopoeia.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eTotal hydroethanolic extracts of the different organs (leaves, bulb and rhizomes) were obtained by maceration. Qualitative phytochemistry of the extracts was carried out using staining methods to identify the main chemical groups. Total flavonoids were quantified by spectrophotometric assay. Two methods were used to assess antioxidant activity: the phosphomobyldate reduction method and the FRAP test. \u003cem\u003ein vitro\u003c/em\u003e antimicrobial tests were carried out using the solid-state diffusion method (Puits et disque).\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eThe highest hydroethanol extraction yield was obtained with the \u003cem\u003eA. difformis\u003c/em\u003e leaf bark extract (9.47%). The results showed the presence of alkaloids, coumarins and reducing sugars. The highest content of total flavonoids was found in the bulbs, with 33.78 \u0026micro;g Eq routine/mg DM. The plant leaves showed high antioxidant activity in vitro with 37.79 EAA/g DM and 270.50 \u0026micro;mol Eq FeSO4/mg DM. On completion of the antimicrobial tests, there was no visible inhibition zone diameter for all the extracts with the two methods used.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003eThe present study invalidates the local use of this plant in pathologies caused by the microbial strains studied. However, research into other activities associated with this plant could explain its use in traditional medicine.\u003c/p\u003e","manuscriptTitle":"in vitro phytochemical and biological studies of the hydroethanolic extract of Anchomanes difformis used in phytotherapy in Togo","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-08-26 07:40:10","doi":"10.21203/rs.3.rs-7401969/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-09-30T11:30:37+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-28T14:58:18+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"195272877410889106160610506111401693483","date":"2025-09-17T07:16:20+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-16T22:16:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"176502581123789138645658262649037286156","date":"2025-09-14T17:20:44+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-10T23:32:10+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"316511783747299558314879363943421159014","date":"2025-09-03T22:52:36+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"282030065594968096451218027768317046876","date":"2025-09-02T06:49:27+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-01T15:52:10+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-01T15:48:07+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-09-01T11:52:34+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-08-23T10:59:00+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-08-23T10:55:47+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"db6633bd-9fcf-4d9d-a00e-bd869136d8e6","owner":[],"postedDate":"August 26th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":53606127,"name":"Biological sciences/Biochemistry"},{"id":53606128,"name":"Biological sciences/Biological techniques"},{"id":53606129,"name":"Biological sciences/Drug discovery"},{"id":53606130,"name":"Biological sciences/Microbiology"},{"id":53606131,"name":"Biological sciences/Plant sciences"}],"tags":[],"updatedAt":"2025-12-01T16:08:07+00:00","versionOfRecord":{"articleIdentity":"rs-7401969","link":"https://doi.org/10.1038/s41598-025-30054-w","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2025-11-28 15:57:27","publishedOnDateReadable":"November 28th, 2025"},"versionCreatedAt":"2025-08-26 07:40:10","video":"","vorDoi":"10.1038/s41598-025-30054-w","vorDoiUrl":"https://doi.org/10.1038/s41598-025-30054-w","workflowStages":[]},"version":"v1","identity":"rs-7401969","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7401969","identity":"rs-7401969","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}