Antibacterial parameters of infused extracts of Mangifera indica cultivars (Kent) leaves on the in vitro growth of urinary tract infections strains

 Corresponding author: Monon KONE Copyright © 2025 Author(s) retain the copyright of this article. This article is published under the terms of the Creative Commons Attribution License 4.0. Antibacterial parameters of infused extracts of Mangifera indica cultivars (Kent) leaves on the in vitro growth of urinary tract infections strains Kpindotchin Landry COULIBALY 1, Monon KONE 1, *, Mani Adrien KOUANGBE 2 and Mohamed CISSE 1 1 Department of Biochemistry-Genetics, UFR Sciences Biologiques, Université Peleforo Gon Coulibaly, BP 1328 Korhogo, Korhogo, Côte d'Ivoire. 2 Department of Microbiology and Molecular Biology, UFR Agriculture, Ressources Halieutiques et Agro-industrie, Université de San Pedro, BP 1800 San Pedro, San Pedro, Côte d'Ivoire. World Journal of Advanced Research and Reviews, 2025, 28(02), 1230-1237 Publication history: Received on 25 September 2025; revised on 08 November 2025; accepted on 12 November 2025 Article DOI: https://doi.org/10.30574/wjarr.2025.28.2.3564 Abstract Background: Mangifera indica is a plant belonging to the Anacardiaceae family. It is traditionally used to treat a variety of conditions including diabetes, bronchitis, diarrhea, asthma, scabies, respiratory problems and urinary tract infections. The aim of the present work is to understand the use of Kent leaves in the treatment of urinary tract infections in traditional medicine. Methods: In this study, qualitative phytochemical tests based on detection by staining and tube precipitation of phenolic compounds were carried out. Bacterial susceptibility to infusions was determined by the agar diffusion method from wells. MICs and BMCs were determined by double dilution in liquid medium coupled with spreading on Mueller Hinton agar. Results: This work shows a dominance of sterols, polyterpenes, polyphenols, flavonoids, catechic tannins, alkaloids and quinones in infusions extracts of kent cultivar. The most sensitive strain was the one with the lowest MIC (MIC = 3.125mg/mL) and the largest diameters (19.33 mm). This was S. aureus 9044. All the germs studied were resistant to at least two families of antibiotics. They are therefore multi-resistant. Conclusion: These results could stimulate the use of kent cultivar leaves in traditional environments for the treatment of urinary tract infections. Keywords: Mangifera Indica; Phytochemical; Antibacterial; Multi-Resistant; Urinary Infection 1. Introduction Health is of paramount importance to man. To maintain good health, man has always resorted to medicines. However in recent years, the emergence and spread of antibiotic resistance represents a real threat to global public health [1]. The situation is alarming in countries where infectious diseases, poverty and malnutrition are endemic. Moreover, the abusive use of antibiotics, contributes to the development of bacterial resistance [2]. The emergence of bacterial resistance is a complex process, often involving host and pathogen environmental factors [3]. Faced with the limitations of available antibiotics, it is essential to search for new bioactive substances with a broad spectrum of action. Medicinal plants are an important source of bioactive molecules that could be exploited in the therapy of infectious diseases such as urinary tract infections [4], as shown by several ethno pharmacological studies [5]. Urinary tract infections are more common in infants and pregnant women. According to the World Health Organization, urinary tract infections (UTIs) are a health security problem for countries, especially those in the developing world [6]. Urinary tract infections are implicated in the onset or complication of serious chronic diseases with a poor prognosis, such as prostatitis, diabetes, World Journal of Advanced Research and Reviews, 2025, 28(02), 1230-1237 1231 sickle cell disease, renal failure and HIV/AIDS [7-9]. Several bacteria are implicated in urinary tract infections: Escherichia coli (80%), Proteus mirabilis, klebsiella spp, Staphylococcus aureus, Pseudomonas aeruginosa, and many others [10]. Indeed, several plants are used in the treatment of infectious diseases. Among these is the mango tree (Mangifera indica) [11], a member of the Anacardiaceae family. Magnifiera indica is widely distributed throughout the world and includes several cultivars (Amélie, Kent...). In Côte d'Ivoire, the dominant cultivar is Kent. It accounts for over 95% of export production [12]. In addition to its famous fruits, the mango tree has many medicinal properties. Traditionally, extracts from the stem barks, leaves are used to treat various ailments such as diabetes, bronchitis, diarrhea, asthma, scabies, respiratory problems [13]. Despite the many therapeutic virtues possessed by the mango tree, the scientific literature remains insufficiently informed about the use of Kent cultivar leaves in traditional medicine. The aim of the present work is to understand the use of Kent leaves in the treatment of urinary tract infections in traditional medicine. 2. Materials and methods 2.1. Materials 2.1.1. Biological material 2.1.2. Plant material The plant material consists of leaves of Mangifera indica cultivar Kent. The leaves were harvested in August 2023 at Korhogo in the Poro region of northern Côte d'Ivoire. 2.1.3. Bacterial strains tested Three bacterial strains of clinical origin including E. coli (8312), S. aureus (9044), P. aeruginosa (8613) supplied by the Laboratoire d'analyse du CHR de Korhogo and three reference strains E. coli ATCC25922, S. aureus ATCC 19213, P. aeruginosa ATCC27853 from IPCI. 2.1.4. Reagents and chemicals Several reagents and chemicals were used. Muller-Hinton agar, nutrient agar, nutrient broth, distilled water and physiological water (NaCl 0.9%) were used for antibacterial tests. The main developers and reagents listed below were used. Dragendorff and Bouchardât reagents were used to reveal alkaloids. Iron III chloride (FeCl3) 2% was used to reveal polyphenols, tannins and phenolic acids. Hydrochloric alcohol was used for flavonoids. Sterols and terpenes were revealed using acetic anhydride. Quinones were detected using ammonia. 2.2. Methods 2.2.1. Leaf processing Leaves were harvested in August 2023 in Korhogo and dried at room temperature for 3 days. The dried leaves were ground using an electric grinder, and the fine powder obtained was stored in jars in a dry place protected from light and moisture until use. 2.2.2. Preparation of total extracts Total extracts were prepared using the infusion method described by Nogaret and Ehrhart, (2011)14, with slight modifications. This involves soaking the drug, which has been wrapped in blotting paper, in boiling water. The container is covered and the drug is left to infuse for 5 minutes before being removed. After cooling and filtration, the various filtrates are placed in an oven at 50°C for 3 days. In this study, different extracts were prepared by varying the mass. Thus, for the infused extract (1%), 1 g of plant powder is soaked in 100 mL of boiling water. 2.2.3. Phytochemical screening Phytochemical screening tests were based in part on qualitative analysis, using tube staining and precipitation reactions [15]. World Journal of Advanced Research and Reviews, 2025, 28(02), 1230-1237 1232 2.2.4. Evaluation of antibacterial activity Checking the purity of bacterial species To check the purity of bacterial species, strains were streaked on specific culture media TBX (for E. coli); Chapman agar (for S. aureus) and King A (for P. aeruginosa). Plates were incubated at 37°C (for S. aureus and P. aeruginosa) and 44°C (for E. coli) for 24 h. The presence of blue-green colonies on TBX was indicative of E. coli; yellow colonies on Chapman indicated S. aureus and green colonies on King A were indicative of P. aeruginosa. Inoculum preparation Using a pasteur pipette, an 18 h colony is picked and placed in a test tube containing 10 mL of sterile Muller-Hinton broth. The mixture is incubated at 37°C for 3 hours. After opalescence, a 0.1 mL suspension of this pre-culture is taken and diluted in 10 mL sterile Muller-Hinton broth, then homogenized. This solution constitutes the stock solution at 100 containing 106 cells/mL [16]. Inoculum counting Inoculum counting is carried out by diluting pure inoculum to the 10th. The various dilutions obtained, together with the pure inoculum, are inoculated onto Mueller Hinton agar without antimicrobial using a calibrated loop of 2 μL per 5 cm long streak. This plate is called plate A or bactericidal control plate. It is incubated at 37°C for 24 h [17]. Preparation of the extract concentration range The plant extract concentration range from 200 mg/mL to 1.56 mg/mL was prepared in test tubes using the double dilution method. The prepared range was autoclaved at 121°C for 15 min [18]. Seeding of the concentration range of extracts Seeding of the concentration range is carried out by adding 1 mL of the contents of each concentration range tube to 1 mL of inoculum at dilution 100 containing 106 cells/mL, around the flame of the Bunsen burner. The Growth Control (TC) tube contains 2 mL of inoculum. The Sterility Control (SC) tube contains 2 mL of sterile culture medium. Susceptibility test Sensitivity of strains to plant extracts was carried out using the agar diffusion technique. Mueller Hinton medium was inoculated by swabbing. Using a sterile punch, wells approximately 6 mm in diameter were made in the agar. Each well received 80 µL of the test substance. After 15 min diffusion at laboratory temperature, the Petri dishes were incubated at 37°C for 24 h. The presence of a zone of inhibition was observed and interpreted according to Ponce et al. (2003)19. 2.2.5. Determination of antibacterial parameters Minimum Inhibitory Concentration (MIC) The MIC is determined from the seeded concentration range by observation of the test tubes with the naked eye. A cloudy medium means that the culture is growing, and a clear medium means that growth has been inhibited. The MIC corresponds to the lowest inoculated concentration without growth visible to the naked eye [16]. Minimum Bactericidal Concentration (MBC) The MBC is determined by subculturing all the experimental tubes in the seeded concentration range without growth visible to the naked eye. This subculture constitutes box B. The BMC is defined by comparing Box A and Box B. It corresponds to the smallest concentration in Box B whose colony count is less than or equal to the colony count of the 10-4 dilution of Box A [16]. 2.2.6. Preparing the antibiogram Preparing the bacterial inoculum The inoculum was prepared from a pure culture. One or two 18h bacterial colonies were picked with a pasteur pipette and emulsified in a tube containing 2mL of 0.85% NaCl physiological water (Bio Mérieux, reference 08026 E). This mixture was homogenized by vortexing and the suspension was calibrated to the 0.5 Mac Farland scale. The inoculum was obtained by adding 100 µL of the previous mixture to 10 mL of sterile physiological water in a screw-top tube [16]. World Journal of Advanced Research and Reviews, 2025, 28(02), 1230-1237 1233 Inoculation of Mueller-Hinton agar Agar poured into 120 mm-diameter petri dishes was inoculated by swabbing. The plate was dried at 37°C for 5 minutes. Antibiotic discs were applied to the agar surface with forceps, spaced 2 cm apart. Once applied, the antibiotic disc is not moved. The plates are incubated for 24 hours at 37°C for all strains. After incubation, the different diameters of the zones of inhibition obtained around the antibiotic discs were measured and interpreted as Susceptible (S), Intermediate (I) or Resistant (R) according to the criteria defined by EUCAST/CA-SFM, 2023. 2.3. Statistical analysis Statistical analyses of the results were carried out using Statistica software version 7.1. Fisher's minimum significant difference (LSD) test was used to determine significant differences between several means. Differences were considered significant at the 5% level. 3. Results 3.1. Extraction yields Extraction yields are shown in Table 1. The highest yields were obtained with the 1% infusates (25.28 ± 1.42). The lowest yields were obtained with the 5% infusions (21.56 ± 0.83). The lowest infusion percentage (1%) recorded the highest extraction yields. Infused extracts from the Kent cultivar showed a sticky appearance and brown coloration, except for the 5% infused which showed a brown coloration. Statistical analysis indicated that there was a significant difference between extraction yields at p ≤ 0, 05. 3.2. Phytochemical screening Phytochemical screening showed the presence of sterols, polyterpenes, polyphenols, flavonoids, catechic tannins, alkaloids and saponins in all three Kent extracts. The difference in phytochemical compounds between the three infused extracts is observed in quinones and gall tannins. The 1% and 2.5% extracts contain gall tannins but no quinones, while the 5% extract shows the presence of quinones but no gall tannins (Table 2). Table 1 Extraction yields for the Kent variety Infused Yield (%) Color Appearance 1% 25,28 ± 1 ,42a Brown Tights 2,5% 24 ,67 ±1,25a Brown Tights 5% 21,56 ± 0,83b Brown Tights Data are expressed as mean ± standard deviation (3 trials). Averages assigned the same letter in the same column are not statistically different at the 5% threshold Table 2 Phytochemical screening Extracts Sterols and Polyterpenes Polyphenols Flavonoides Tannins Quinones Alcaloides Saponines Cat Gal B D K.1% + + + + + - + + + K.2,5% + + + + + - + + + K.5% + + + + - + + + + (-) : absence ; (+) : presence ; K : Kent ; Cat. : Catechique ; Gal. : Galliques ; B : Bouchardât ; D : Dragendorff 3.3. Abacterial activity 3.3.1. Germ Susceptibility The results of bacterial sensitivity testing on extracts are shown in Table 3. The largest inhibition diameters were obtained at a concentration of 200 mg/mL. The K.2.5% infusion obtained the best inhibition diameters. The largest World Journal of Advanced Research and Reviews, 2025, 28(02), 1230-1237 1234 inhibition diameter (19.33 ± 0.94 mm) was obtained with the S. aureus strain (9044) and the smallest with E. coli (8312) and E. coli ATCC 25922. For the 5% infusion, the largest inhibition diameter (16.66±1.67 mm) was obtained with P. aeruginosa (8613). As for the 1% infusion, the largest inhibition diameter was obtained with S. aureus strain 9044 (14.67±0.47 mm). All germs were sensitive to all three extracts except E. coli (8312) and E. coli ATCC25922, which showed no inhibition diameter. S. aureus was the most sensitive strain to the different extracts, and E. coli (8312) the least sensitive. Statistical analysis indicates that for each germ, there is no significant difference between the mean inhibition diameters obtained with K.1% and K.5% infusions at p≤0.05. 3.3.2. Determination of antibacterial parameters (MIC and MBC) All extracts showed minimum inhibitory concentrations (MIC) ranging from 3.125 to 12.5 mg/mL. Minimum bactericidal concentrations (MBC) ranged from 0 to 50 mg/mL for the three (3) infused extracts (Table 4; 5 and 6). The 1% and 5% infusions exerted a bacteriostatic effect on P. aeruginosa and S. aureus strains, while the 2.5% infusion had a bactericidal effect on S. aureus strains and a bacteriostatic effect on P. aeruginosa strains. Table 3 Inhibition diameter of extracts Inhibition diameter of the concentration of 200 mg/mL Origines Germes K.1% K.2.5% K.5% Gen. 30.103 mg Cefo. 5.103 mg Souches cliniques E. coli (8312) 0a 0a 0a 9 0 S. aureus (9044) 14,67±0,58b 19,33±1,15c 15,33±0,58b 14 12 P. aeruginosa (8613) 14±1,73d 14,67±2,51d 16,66±2,08d 13 8 Souches de references E. coli ATCC25922 0e 0e 0e 21 13 S. aureus ATCC19213 14,5±0,50f 15,5±0,50f 15±0,00e 20 11 P. aeruginosa ATC27853 14,5±1,50g 15±0,00g 16±1,00g 23 7 Data are expressed as mean ± standard deviation (3 trials). Averages assigned the same letter in the same column are not statistically different at the 5% threshold Table 4 Kent 1% antibacterial parameters Paramètres antibactériens de l’extrait Origines Germes CMI (mg/mL) CMB (mg/mL) CMB / CMI Interprétation Souches cliniques E. coli 8312 Nd Nd Nd Nd S. aureus 9044 3,125 50 16 Bactericidal P. aeruginosa 12 ,5 50 4 Bacteriostatic Souches de références E. coli ATCC25922 Nd Nd Nd Nd S. aureus ATCC 19213 3,125 50 16 Bacteriostatic P. aeruginosa ATCC27853 12,5 50 4 Bacteriostatic Nd : not determined World Journal of Advanced Research and Reviews, 2025, 28(02), 1230-1237 1235 Table 5 Kent 2.5% antibacterial parameters Paramètres antibactériens de l’extrait Origines Germes CMI (mg/mL) CMB (mg/mL) CMB / CMI Interprétation Souches cliniques E. coli 8312 Nd Nd Nd Nd S. aureus 9044 3,125 3,125 1 Bactericidal P. aeruginosa 12 ,5 50 4 Bacteriostatic Souches de références E. coli ATCC25922 Nd Nd Nd Nd S. aureus ATCC 19213 3,125 6,25 2 Bactericidal P. aeruginosa ATCC27853 12,5 50 4 Bacteriostatic Nd : not determined Table 6 Kent 5% antibacterial parameters Paramètres antibactériens de l’extrait Origines Germes CMI (mg/mL) CMB (mg/mL) CMB / CMI Interprétation Souches cliniques E. coli 8312 Nd Nd Nd Nd S. aureus 9044 3,125 12,5 4 Bacteriostatic P. aeruginosa 12 ,5 50 4 Bacteriostatic Souches de références E. coli ATCC25922 Nd Nd Nd Nd S. aureus ATCC 19213 3,125 50 16 Bacteriostatic P. aeruginosa ATCC27853 12,5 50 4 Bacteriostatic Nd : not determined 3.4. Resistance profile of the bacteria studied The antibiotics studied have been grouped into 4 families (Aminosides, Penicillins, Cephalosporins and Beta-lactamins). Each antibiotic family contains two antibiotics tested, with the exception of the Beta-lactam family, which has a single antibiotic (Ceftazidine). Results were interpreted according to CASFM, 2023. All germs were resistant to gentamicin (CN-30mcg), amikacin (AK-30µg), amoxicillin (AX-30mcg), amoxicillin + Ac. clavulanique (AUG.30 µg), ceftriaxome (CRO-30 µg), cefotaxime (CTX 5µg), ceftazidine (CAZ 30µg) except E. coli (8312) which was sensitive to the antibiotic Amikacin (AK-30µg). 4. Discussion Bioactive molecules present in plant species are very important in the treatment of certain pathologies. In this work, we aimed to contribute to an in-depth understanding of the therapeutic potential of mango leaves from the Kent cultivar. The highest extraction yields were obtained with 1% infusions and the lowest with 5% infusions. Statistical analysis shows a significant difference between extraction yields. This difference can be explained by the solid/liquid ratio. The higher the solvent volume, the greater the degree of contact between the drug and the extraction solvent. This increases the solvent's capacity to penetrate the drug, enabling the extraction solvent to come into contact with a large number of compounds. These results are similar to those of Koné et al. (2017)20, who showed that yields are higher when the volume/mass ratio of the grind is high. Phytochemical screening showed the presence of sterols, polyterpenes, polyphenols, flavonoids, catechic tannins, alkaloids and saponins in infused extracts of the Kent cultivar. The presence or absence of quinones and gall tannins differentiates the composition of the extracts. The K.1% and K.2.5% infusions have the same phytochemical composition, but differ from that of K.5%. The absence of quinones in K.1% and K.2.5% infusions may be explained by less efficient extraction at lower concentrations. Conversely, the absence of gall tannins in the K.5% infusion may be due to complexation, precipitation or chemical interaction with other compounds at higher World Journal of Advanced Research and Reviews, 2025, 28(02), 1230-1237 1236 concentrations, limiting their availability in the final extract. These observations are consistent with the work of Sarker and Nahar (2012)21, who studied the effects of extraction methods and chemical interactions on the phytochemical composition of plant extracts. Our results corroborate those of Mustapha et al. (2014)22. These authors found the same phytochemical compounds in mango leaves harvested in Nigeria. However, the literature has reported that Magnifera indica leaves from Burundi harvested at different times and sites contained no alkaloids [23]. These results show that the phytochemical composition of medicinal plant leaves can vary according to the time of harvest, the nature of the soil, the age of the plant and the drying conditions of the organ studied [24]. The active ingredient content of Mangifera indica leaves is comparable to that previously found for Psidium guajava leaves [25]. Mangifera indica's richness in these major groups of active chemical compounds could then justify the traditional use of this plant to treat numerous illnesses such as diabetes, bronchitis, diarrhea, asthma, scabies and respiratory problems [26]. With regard to antimicrobial testing, the tests carried out highlighted the growth-inhibiting activity in vitro of E. coli (8312), S. aureus (9044) and P. aeruginosa (8613) by the infused extracts. The strains tested were resistant to at least two antibiotics from two different families. They are therefore described as multi-resistant. The sensitivity of the strains studied to the infused extracts varied from one bacterial strain to another. S. aureus 9044 was the most sensitive strain to Kent 1% and 2.5% infused extracts. As for the Kent 5% infusion, P. aeruginosa (8613) was the most sensitive. E. coli (8312) was the least sensitive strain to the plant extracts studied. Despite the multi-resistance of the strains studied, our extracts achieved interesting inhibition diameters, except on E. coli. Determination of antibacterial parameters reinforced this sensitivity. The 1% and 5% infusions of the Kent cultivar showed a bacteriostatic effect on P. aeruginosa and S. aureus strains, while the 2.5% infusion showed a bactericidal effect on S. aureus strains and a bacteriostatic effect on P. aeruginosa strains. The high presence of phytochemicals, notably tannins, could be at the root of this efficacy. 5. Conclusion This work contributes to the valorization of the medicinal plant Mangifera indica. Extraction by infusion showed that 1% is the best percentage for extracting active ingredients from plant extracts. Phytochemical screening of infused extracts from the leaves of the kent cultivar recealed a richness in sterols, polyterpenes, polyphenols, flavonoïdes, tannins catechiques, alcaloïdes et saponides. The antibacterial activity of the infusions showed promising results despite the multiresistance of the bacteriatested. The 1 % and 5 % infused extracts demonstrated a bacteriostaticeffect on P. aeruginosa and S. aureus, while the 2.5 % infused extract exhibited a bactericidal effect on S. aureus and a bacteriostatic effect on P. aeruginosa. The richness of phytochemical compounds in the infusions could explain the observed biological activities. These results suggest that Mangifera indica could offer hope in the treatment of urinary tract infections, a major public health threat. 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