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. 2026 Sep 29. Online ahead of print. doi: 10.1039/d6ra06543k

Green synthesis of a manganese oxide–selenium nanocomposite using watermelon peel waste with antimicrobial and mosquitocidal activities

Samy Selim a,✉, Abeer S Aloufi b, Tharwat A Selim c, Amr H Hashem d,✉, Mohammed H Alruhaili e,g, Hattan S Gattan f,g, Alaa A Kashmiry h, Nosiba S Basher i, Ebrahim Saied d
PMCID: PMC13620934  PMID: 42812612

Abstract

The present study demonstrates the successful green biosynthesis of a manganese oxide–selenium nanocomposite (MnO2–Se NC) using watermelon peel waste extract, providing a sustainable approach for valorizing agro-industrial waste into a multifunctional nanomaterial. The biofabricated nanocomposite was characterized by UV-Vis, XRD, FTIR, DLS and TEM analysis. TEM analysis revealed predominantly spherical nanoparticles with an average size of approximately 55 nm, while UV-Vis spectra displayed two distinct absorption peaks at 266 and 300 nm, confirming successful nanocomposite formation. The MnO2–Se NC exhibited potent antibacterial activity, with MIC values ranging from 3.9 to 31.25 µg ml−1 and MBC values between 15.62 and 125 µg ml−1. The highest efficacy was observed against Bacillus subtilis (MIC = 3.9 µg ml−1), whereas comparatively lower activity was recorded against Pseudomonas aeruginosa (MIC = 31.25 µg ml−1), with MBC/MIC ratios of 2–4 indicating a bactericidal mode of action. In addition, the nanocomposite demonstrated promising antifungal activity, with MIC/MFC values of 31.25/62.5 µg ml−1 against Candida albicans and 125/500 µg ml−1 against Aspergillus brasiliensis, suggesting greater effectiveness against yeast than filamentous fungi. Furthermore, the MnO2–Se NC showed strong larvicidal and adulticidal activity against Culex pipiens, with significantly lower LC50 and LC90 values (5.6–7.0 ppm) compared to the crude watermelon peel extract (66–100 ppm). These findings indicate enhanced toxicity and efficacy of the bimetallic nanocomposite at substantially lower concentrations. In conclusion, the findings highlight the considerable multifunctional potential of watermelon peel-derived MnO2–Se NC as a sustainable antimicrobial and vector-control agent. The combined biological activities, together with the use of an abundant agricultural waste resource for synthesis, support its future development as an environmentally friendly nanomaterial for biomedical and mosquito-management applications.

The present study demonstrates the successful green biosynthesis of a manganese oxide–selenium nanocomposite using watermelon peel waste, providing a sustainable approach for valorizing agro-industrial waste into a multifunctional nanomaterial. Image designed using Google Gemini.graphic file with name d6ra06543k-ga.webp

Introduction

Nanomaterials, particularly nanocomposites, have attracted considerable attention because their nanoscale dimensions confer distinctive physicochemical and biological properties compared with their bulk counterparts.1,2 Nanocomposites are formed by integrating two or more nanoscale components, which can generate synergistic properties and enhance biological and functional performance compared with individual nanomaterials.3,4 Bimetallic nanocomposites have therefore emerged as promising multifunctional materials for applications in biomedicine, agriculture, and environmental protection.4 However, conventional physical and chemical approaches for nanoparticle synthesis often require hazardous reagents, high energy consumption, and complex processing, raising environmental and biological concerns.4,5 Consequently, increasing attention has been directed toward green synthesis strategies that are safer, more economical, and environmentally sustainable.6,7 Plant extracts are particularly attractive for this purpose because their phytochemicals and secondary metabolites can act as natural reducing, capping, and stabilizing agents during nanoparticle formation.8

Selenium has recently attracted attention in its nanoparticulate form, owing to its crucial role in human health and nutrition, as well as its notable bioactivity and relatively low toxicity.9 Despite these advantages, selenium nanoparticles (SeNPs) often suffer from instability and a tendency to aggregate during practical applications, highlighting the need for straightforward and reliable approaches to ensure their dispersion and stabilization.10 Green synthesis via biological reduction using plant extracts has been suggested as a feasible strategy for the preparation of SeNPs, offering advantages of simplicity, safety, and scalability.11 To date, various plant systems have been employed in SeNP biosynthesis, such as ginger extract, fenugreek seed extract, Clausena dentata leaf aqueous extract, and Vitis vinifera (raisin) extract.12–15

Manganese is also an essential trace element involved in several important biological processes, including photosynthesis, respiration, enzymatic activity, reactive oxygen species detoxification, and hormone signaling.16,17 Manganese-based nanomaterials may provide an efficient form of manganese and have demonstrated potential for improving stress tolerance compared with conventional forms.18 The combination of manganese oxide and selenium in a single nanocomposite could therefore provide complementary or synergistic properties, potentially enhancing its biological effectiveness.

Mosquito-borne diseases remain a major public-health concern, with Culex species representing important vectors of several pathogens. Culex pipiens, which is widely distributed in Egypt, has been associated with the transmission of West Nile virus, Wuchereria bancrofti, and Rift Valley fever virus.19–22 Effective mosquito control is therefore essential for reducing vector-borne disease transmission. However, extensive reliance on synthetic insecticides, including organochlorine and organophosphate compounds, has raised concerns regarding environmental pollution, toxicity to humans and non-target organisms, and reduced effectiveness associated with insecticide resistance.23 Consequently, plant-derived materials and biologically synthesized nanoparticles have emerged as promising alternatives because of their potential cost-effectiveness, environmental compatibility, and biological activity.24,25 Bimetallic nanoparticles may exhibit enhanced toxicity compared with their individual components, although their effectiveness can vary according to nanoparticle composition, physicochemical characteristics, mosquito species, and the reducing and stabilizing agents involved in their synthesis.26

Agricultural and food-processing wastes represent abundant and inexpensive sources of phytochemicals that can be exploited for the sustainable synthesis of nanomaterials. Watermelon peel waste, in particular, represents an underutilized biomass resource that may provide suitable bioactive compounds for nanoparticle reduction and stabilization. Despite the increasing interest in plant-mediated nanoparticle synthesis, the green biosynthesis of a manganese oxide–selenium nanocomposite (MnO2–Se NC) using watermelon peel waste extract has, to the best of our knowledge, not previously been reported. Therefore, this study aimed to develop an environmentally friendly strategy for the green synthesis of a MnO2–Se NC using watermelon peel waste extract as a natural reducing and stabilizing agent for the first time, and to evaluate its physicochemical characteristics along with its antimicrobial, antifungal, and mosquitocidal activities to explore its potential as a sustainable multifunctional nanomaterial for biomedical and vector-control applications.

Materials and methods

Preparation of WPW extract

Watermelon peel waste (WPW) was collected from local marketplaces. The preparation of the WPW aqueous extract was performed according to the method previously reported by Patra, Baek,27 with some modifications. The white portion of the watermelon peel was separated, thoroughly washed with deionized water, and cut into approximately 1 cm pieces. Subsequently, 200 g of the prepared peel was transferred into a 2 L conical flask containing 1000 ml of deionized water. The mixture was heated and continuously stirred using a magnetic stirrer for 10 min to facilitate the extraction of water-soluble phytochemicals. The resulting aqueous extract was then filtered through Whatman no. 1 filter paper to remove residual plant material. The filtrate was collected in a sterile container and stored at 4 °C until further use in the biosynthesis process.

Green biosynthesis of MnO2–Se nanocomposite

The MnO2–Se NC was synthesized using the WPW aqueous extract as a natural reducing and stabilizing agent. Briefly, 20 ml of Na2SeO3 solution (2 mM) and 20 ml of Mn(CH3COO)2·4H2O solution (2 mM) were mixed with 60 ml of WPW extract, giving a final reaction volume of 100 ml. The reaction mixture was rapidly stirred and maintained at 60 °C. The pH of the reaction mixture was adjusted using 1 N NaOH. The mixture was continuously stirred at 60 °C for 2 h. The formation of the MnO2–Se nanocomposite was preliminarily indicated by a distinct color change of the reaction mixture from pale yellow to dark reddish-brown. After completion of the reaction, the formed precipitate was collected by centrifugation, followed by drying in a hot-air oven at 160 °C for 6 h. The resulting dried material was subsequently subjected to physicochemical characterization. The biosynthesis procedure was performed according to ref. 28, with the conditions described above.

The formation of the MnO2–Se nanocomposite can be represented schematically by the following reactions:

Mn(CH3COO)2·4H2O + 2NaOH → Mn(OH)2↓ + 2CH3COONa + 4H2O 1
2Mn(OH)2 + O2 → 2MnO2 + 2H2O 2
SeO32− + 3H2O + 4e− → Se0 + 6OH− 3
Mn(CH3COO)2 + NaOH + Na2SeO3 + WPW phytochemicals → MnO2–Se NC 4

Accordingly, the phytochemicals present in the WPW extract may contribute to the reduction of selenium species and stabilization of the newly formed nanostructures, while manganese hydroxide undergoes conversion toward manganese oxide during the synthesis process. The exact reaction pathway may involve several intermediate steps depending on the phytochemical composition of the WPW extract and the reaction conditions.

Characterization of MnO2–Se NC

The formation of the MnO2–Se NC was initially monitored visually by observing the characteristic color change of the reaction mixture from pale yellow to dark reddish-brown. The synthesized nanocomposite was subsequently characterized using complementary physicochemical techniques. UV-Vis spectroscopy: The optical absorption spectrum of the synthesized MnO2–Se NC was recorded over the wavelength range of 200–700 nm to detect the characteristic absorption behavior associated with nanocomposite formation. Fourier-transform infrared spectroscopy (FTIR): FTIR analysis was performed using the KBr pellet technique over the range of 400–4000 cm−1 to identify the functional groups present on the surface of the synthesized nanocomposite and to assess their possible involvement in nanoparticle formation and stabilization.

Transmission electron microscopy (TEM): TEM analysis was conducted using a JEM-2100 Plus instrument to determine the morphology, shape, dispersion, and particle size of the synthesized MnO2–Se NC. Dynamic light scattering (DLS): the hydrodynamic particle size distribution of the synthesized nanocomposite was determined using a Nano ZS (Malvern) analyzer. X-ray diffraction (XRD): XRD analysis was carried out using an XRD-6000, Shimadzu diffractometer to determine the crystalline structure and crystallinity of the synthesized MnO2–Se NC. Collectively, these complementary techniques were used to confirm the successful green synthesis of MnO2–Se NC and to evaluate its optical, chemical, morphological, particle-size, and crystalline characteristics.

Antibacterial activity assay

The antibacterial activity of the synthesized MnO2–Se NC was evaluated by agar well diffusion method toward Bacillus subtilis ATCC 6051, Staphylococcus aureus ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and Escherichia coli ATCC 25922. Fresh bacterial solutions (about 106 CFU ml−1) were calibrated and evenly spread on the plates of Mueller–Hinton agar. Sterile agar wells (7 mm diameter) were filled with MnO2–Se NC (500 µg ml−1), sodium selenite (500 µg ml−1), manganese acetate (500 µg ml−1), and levofloxacin as positive controls. After 24 h of incubation, Inhibitory zones were measured in mm. The MIC was measured by successive two-fold dilutions of MnO2–Se NC in Mueller–Hinton broth. 24 h of incubation at 37 °C after inoculation with bacterial suspension, optical density (OD) was measured using ELIZA reader at 600 nm. To calculate MBC, aliquots from MIC tubes that showed no growth were plated on fresh agar plates and grown under the same conditions. The MBC was the lowest quantity that prevent bacterial colony formation.

Antifungal activity assay

Antifungal activity of MnO2–Se NC was evaluated against Candida albicans ATCC 90028 and Aspergillus brasiliensis ATCC 16404 utilizing agar well diffusion. Fungal solutions were prepared in sterile saline and spores per millilitre were quantified. Suspensions were evenly spread on potato dextrose agar (PDA) plates, MnO2–Se NC (500 µg ml−1), precursor metals (500 µg ml−1) and fluconazole (positive control) were added to wells (7 mm) separately. Inhibitory zones were measued after 48–72 h at 28 °C. MIC was evaluated by broth microdilution method with MnO2–Se NC diluted in PDB medium. Sub-cultured samples from the growth-inhibited wells were incubated on fresh PDA plates to determine the minimal fungicidal concentration (MFC). The lowest dose of the MFC fully prevent the growth of the fungal colonies.

Mosquito rearing technique

Culex pipiens was obtained from the Medical Entomology Institute. It was reared for several generations, under controlled laboratory conditions at the temperature of 27 ± 2 °C, relative humidity 70 ± 10% and 12–12 light–dark regime.

Larvicidal activity

The larvicidal activity of MnO2–Se NCs and WPW extract was assessed against C. pipiens mosquitoes using 21's technique. Twenty larvae of the species under investigation were selected from the early third instar. To make it easier to compare the results with those of other nanoscale substances, the WPW extract was evaluated at concentrations ranging from 100 to 500 ppm. MnO2–Se NC, on the other hand, was added in 10–50 ppm increments. For every concentration, three tests were carried out. Under the same experimental circumstances, all extract treatments were evaluated simultaneously with a control group of twenty larvae in 250 milliliters of tap water. Twenty-four hours after therapy, the death % was calculated.

Adulticidal activity

Twenty female mosquitoes were moved from the colony to a plastic tube (holding section) for an hour in order to evaluate the adulticidal efficiency. The larvae were then moved to the exposure compartment on the other side, which had a Whatman no. 1 filter paper sheet measuring 10 by 10 cm. WPW and MnO2–Se NC were applied to the filter sheets.25 The adult was subjected to the identical concentrations of WPW and MnO2–Se NC extracts based on the larvicidal properties. Following the exposure period, the adults were gently placed back in the holding section after being given a cotton bag soaked in a 10% sucrose solution and placed on top of the mesh net. For every concentration, three replicates were carried out. As stated in reference the mortality of adult was assessed and the fatal dosages LC50 and LC90 were ascertained by probit analysis.29

Statistical analysis

Descriptive statistics such as the mean and standard deviation (SD) were computed for each treatment. Probit analysis was used to get LC50, and LC90 with a 95% confidence interval based on the average larval mortality data. The statistical analysis performing a one-way analysis of variance. The software used for this study was SPSS (version 25). The LSD post hoc test was used for pairwise comparisons.

Results and discussion

Biosynthesis of MnO2–Se NC

Biological resources, including plant extracts or biomass, offer an environmentally friendly to replace conventional physical and chemical techniques used for the safe, fast and inexpensive manufacture of NCs.30 In the present study, WPW extract served as the main reducing and stabilizing agent for biosynthesis of MnO2–Se NCs. During the reduction process, the reaction mixture exhibited a dark reddish-brown coloration, indicating the formation of new NCs. This color transition reflects the capacity of bioactive compounds in the extract to interact with metal ions and drive their conversion into MnO2–Se NCs.31

Similar results were also reported by Elkady et al.,28 where they synthesized and characterized copper oxide–selenium nanoparticles using Lagenaria siceraria leaf extract. Similarly,32 showed the production of selenium–gold BNPs from Pluchea indica leaves, also Selim et al.33 fabricated silver–titanium dioxide nanoparticles from Pluchea indica leaves confirming their anticancer, antimicrobial, and antioxidant activities.

Saied et al.34 prepared bimetallic TiO2–ZnO nanoparticles from onion peels with antibacterial, antibiofilm and anticancer properties. Kumar et al.35 synthesized bimetallic Au–Pt NPs from Croton caudatus leaf extract. Mirzaei et al.36 developed silver selenide nano-chalcogens using aqueous extract of Melilotus officinalis, while Sultana et al.37 prepared selenium–iron nanocomposite from Allium sativum extract. In addition, Hashem et al.38 reported the use of watermelon peel extract to biosynthesize selenium–silver BNPs, which showed strong antimicrobial and antioxidant activities.

UV-visible spectroscopy

UV-visible spectroscopy is a widely used method for assessing the optical properties of nanomaterials. In this study, MnO2–Se NC was synthesized using WPW extract, and its optical properties were examined within the wavelength range of 200–700 nm. The UV-Vis spectrum revealed two main absorption bands: a broad band centred at 300 nm, associated with Se0-related absorption, and another band at 266 nm, attributed to MnO2-related electronic transitions (Fig. 1). The color change from pale yellow to dark reddish-brown during synthesis further supported the formation of the nanocomposite. This color variation may be associated with electronic transitions and changes in the optical properties induced by nanoparticle formation. Lagenaria siceraria leaf extract was used by Elkady et al.28 to form bimetallic copper oxide–selenium nanoparticles, with the SPR band maximum observed at 330 nm. B2O3–ZnO BNPs exhibited a UV-Vis absorption peak at 370 nm, according to Hashem et al.,39 while selenium–silver BNPs showed a characteristic band at 380 nm.43 Monometallic CuO NPs and MnO NPs exhibited surface plasmon resonance maxima at 336 nm and 266 nm, respectively.40

Fig. 1. UV-vis spectra of Biosynthesized MnO2–Se NC and WPW extract.

Fig. 1

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Furthermore, Salem et al.41 observed a distinct absorption band at 295 nm for SeNPs synthesized from orange peel waste. Similarly, Hussein et al.42 reported a well-defined absorption band at 285 nm for MnO2 NPs, confirming their formation. Furthermore, Banerjee, Rajeswari43 found that SeNPs synthesized using Moringa oleifera leaves exhibited an absorption band within the 200–400 nm region, with a maximum at 268 nm. In contrast, SeNPs synthesized using tarragon extract showed a ruby-red color and an absorption band near 450 nm Yilmaz et al.,44 while Saod et al.45 reported an absorption peak at 410 nm for MnO NPs in the visible region. These differences in the reported absorption maxima may be attributed to variations in nanoparticle composition, particle size, morphology, surface chemistry, synthesis conditions, and interactions among the constituent components. In the present study, the MnO2–Se nanocomposite exhibited absorption bands at 266 and 300 nm, which were attributed to MnO2- and Se-related electronic transitions, respectively. The observed absorption profile differs from those reported for some individual Se or MnO-based nanoparticles, indicating that the formation of the MnO2–Se nanocomposite can modify its optical response through interactions between the two components. Therefore, the UV-Vis spectrum, together with the observed color change from pale yellow to dark reddish-brown, provides supportive evidence for the formation of the MnO2–Se nanocomposite.

FTIR analysis of MnO2–Se NC

The FTIR spectrum of MnO2–Se NC (Fig. 2) reveals distinct absorption bands associated with phytochemical residues derived from WPW extract as well as metal–oxygen/selenium-related vibrations. A broad band at 3594 cm−1 is assigned to O–H stretching, which may arise from hydroxyl groups of plant-derived compounds and/or adsorbed water molecules.46 The peak at 3094 cm−1 can be associated with C–H stretching vibrations of organic compounds.47 Weak absorption bands at 2525 and 2324 cm−1 may be related to adsorbed CO2 and/or other functional groups present in the organic matrix, suggesting the possible contribution of plant-derived metabolites to the surface chemistry of the synthesized nanocomposite.48 The strong band at 1673 cm−1 is attributed to C Created by potrace 1.16, written by Peter Selinger 2001-2019 O stretching vibrations of carbonyl-containing compounds, while the peak at 1439 cm−1 may be associated with C–N stretching and/or aromatic amine-related vibrations.49 The absorption band at 1110 cm−1 can be assigned to C–O–C stretching vibrations, further indicating the presence of oxygen-containing organic compounds derived from the WPW extract.50 These functional groups, particularly hydroxyl, carbonyl, and ether-containing groups, may participate in the coordination, surface capping, and stabilization of the synthesized nanocomposite and may also contribute to the reduction and transformation of the precursor species during the biosynthesis process.

Fig. 2. FTIR spectrum of MnO2–Se NCs.

Fig. 2

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In the low-frequency region, distinct bands were observed at 768, 594, and 507 cm−1, which may be associated with Mn–O and Se-related vibrations, supporting the presence of the inorganic components of the MnO2–Se nanocomposite.51 In agreement with these findings, the stretching vibrations of Mn–O and Mn–O–Mn bonds have previously been reported at approximately 613 and 570 cm−1, respectively Saod et al..45 Overall, the FTIR results provide supportive evidence for the possible involvement of WPW-derived phytochemicals in the formation and stabilization of the MnO2–Se nanocomposite. The observed changes and characteristic functional groups suggest that plant-derived biomolecules may act as surface-coordinating and capping agents, while the low-frequency metal-related vibrations support the presence of the inorganic components. However, the FTIR findings should be considered supportive rather than definitive evidence of the specific reduction and stabilization mechanisms involved in nanocomposite formation.

XRD of MnO2–Se NCs

The crystalline structure of the biosynthesized MnO2–Se NC was investigated using XRD as shown in Fig. 3. The diffraction pattern of pure MnO2 typically exhibits characteristic peaks at 2θ values of around 28.6°, 37.3°, 42.0°, 56.5°, and 59.9°, which can be indexed to the (110), (101), (200), (211), and (220) planes, respectively, corresponding to the tetragonal crystalline phase of MnO2 (JCPDS card no. 44-0141).52–54 Krishnaraj et al.55 also noted that Ag-doped MnO2 exhibited characteristic diffraction peaks corresponding to planes (111), (200), and (311) for Ag, as well as (100) and (101) for MnO2. Sannasi, Subbian56 reported that the diffraction peaks of MnO2 synthesized using Moringa oleifera gum correspond to the α-MnO2 tetragonal phase with I4/m space group (JCPDS card no. 44-0141). On the other hand, elemental selenium generally shows broad diffraction peaks near 23.5°, 29.7°, 43.6°, and 51.7°, which correspond to the (100), (101), (110), and (201) planes of the trigonal crystalline selenium structure (JCPDS card no. 06-0362).28,57

Fig. 3. XRD spectrum of MnO2–Se NC.

Fig. 3

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In the diffraction pattern of the MnO2–Se NC, distinct peaks corresponding to both MnO2 and Se were observed, confirming the successful synthesis of MnO2–Se NC (Table 1). The presence of characteristic MnO2 peaks together with selenium reflections indicates that Se nanoparticles were well integrated and stabilized on the MnO2 matrix without altering the fundamental crystal structure of either component. Moreover, a noticeable broadening of the diffraction peaks was observed, which can be attributed to the nanocrystalline nature of the material and the small crystallite size. The average crystallite size of the nanocomposite, calculated using the Debye–Scherrer equation, was estimated to be in the range of 56 nm, which is consistent with nanoscale dimensions. Selenium nanoparticles are known for their relatively low crystallinity when synthesized via green methods, which is consistent with the broad and less intense Se peaks observed.58 The integration of Se with MnO2 could result in synergistic effects, particularly in enhancing redox and catalytic activity, as MnO2 provides a stable crystalline matrix while Se contributes to electron transfer processes.59 Similar findings were reported in earlier studies, where incorporation of selenium into metal oxide matrices improved their photocatalytic and antimicrobial activities due to improved charge separation and increased surface area.60 Sultana et al.37 reported that the XRD pattern of green-synthesized Se–Fe NCs showed Bragg's reflections corresponding to selenium (220, 311) and iron (422, 511, 533, 620, 622), matching JCPDS cards no. 06-0362 and 39-1346. Thus, the XRD results confirm the successful green synthesis of MnO2–Se NCs with nanoscale crystallinity, structural stability, and potential functional advantages for environmental applications.

Table 1. XRD peak assignments and Miller indices (hkl) of the MnO2–Se nanocomposite.

2θ (°) Phase Miller index (hkl) Crystallographic reference
28.6 MnO2 (110) JCPDS no. 44-0141
37.3 MnO2 (101) JCPDS no. 44-0141
42.0 MnO2 (200) JCPDS no. 44-0141
56.5 MnO2 (211) JCPDS no. 44-0141
59.9 MnO2 (220) JCPDS no. 44-0141
23.5 Se (100) JCPDS no. 06-0362
29.7 Se (101) JCPDS no. 06-0362
43.6 Se (110) JCPDS no. 06-0362
51.7 Se (201) JCPDS no. 06-0362
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TEM and DLS of MnO2–Se NC

TEM is a powerful tool for characterizing the morphology, particle size, and dispersion of nanoparticles. Fig. 4A shows that the synthesized MnO2–Se NCs consisted of nearly spherical particles with slight agglomeration, while the clearly defined particle boundaries confirmed the successful formation of the nanocomposite. Fig. 4B presents the particle-size distribution obtained by dynamic light scattering (DLS), with a hydrodynamic diameter ranging from approximately 30 to 100 nm and an average diameter of 51 nm.

Fig. 4. TEM (A) and DLS (B) analysis of the synthesized MnO2–Se NC.

Fig. 4

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A comparative analysis of the TEM and DLS measurements was performed to clarify the difference between the particle-size values obtained by the two techniques. TEM determines the physical dimensions of nanoparticles directly from electron micrographs, whereas DLS measures the hydrodynamic diameter of particles dispersed in an aqueous medium. Therefore, the DLS value does not represent only the inorganic nanoparticle core; it also reflects the hydration layer surrounding the particles and the organic molecules adsorbed on their surfaces, including phytochemical constituents derived from the plant extract that may act as reducing, capping, and stabilizing agents. In addition, DLS measurements are sensitive to a small degree of particle association or agglomeration in suspension because larger particles contribute disproportionately to light scattering. Consequently, the hydrodynamic diameter is commonly higher than the physical particle size observed by TEM.61 The slightly higher DLS value obtained in the present study can therefore be attributed to the hydration shell and surface-bound biomolecules, together with limited particle association in the aqueous dispersion, rather than indicating an inconsistency between the two techniques.

The polydispersity index (PDI) was 0.211, indicating a relatively narrow particle-size distribution in the aqueous suspension. PDI values below approximately 0.3 are generally considered indicative of a relatively homogeneous colloidal population. Accordingly, the PDI value of 0.211 suggests that the biosynthesized MnO2–Se NCs exhibited relatively good size uniformity with limited aggregation in aqueous dispersion. The particle-size characteristics observed in the present study were generally comparable with those reported for other green-synthesized nanomaterials. CuO–Se BNPs with an average particle size of 79 nm were previously reported using Lagenaria siceraria leaf extract Elkady et al..28 Similarly, spherical Cu–Mn bimetallic nanoparticles with an average size of 58 nm were reported by Alafaleq et al..40 whereas62 reported Mn–Cu codoped NiO nanorods with dimensions ranging from 30 to 90 nm using Carica papaya leaf extract. These comparable size ranges indicate that plant-mediated synthesis can produce nanomaterials within a similar nanoscale range. In contrast, larger particle sizes ranging from 100 to 150 nm were reported by,63 which may be associated with differences in synthesis parameters, including precursor concentration, reaction temperature, reaction time, pH, and the nature and concentration of the reducing and capping agents. The hydrodynamic size of nanoparticles can vary depending on their composition, surface chemistry, dispersion medium, and experimental conditions.64 The somewhat smaller average hydrodynamic diameter observed for the present MnO2–Se NCs compared with some previously reported nanomaterials may therefore reflect differences in the physicochemical properties of the constituent metal components and the composition of the phytochemicals involved in nanoparticle stabilization. Overall, the combined TEM and DLS results demonstrate that the biosynthesized MnO2–Se NCs possess a nanoscale size with relatively good particle-size uniformity and limited aggregation in aqueous dispersion.

Biological activities of the prepared MnO2–Se NC

Antibacterial activity

In the current study, MnO2–Se NC was assessed for antibacterial activity toward pathogenic bacterial and fungal strains as illustrated in Fig. 5. Also start metals (sodium selenite and manganese acetate) and Lev/Fluc (levofloxacin/fluconazole) were evaluated. Results revealed that the prepared nanocomposite MnO2–Se NC displayed significant antibacterial activity against all tested bacterial strains. MnO2–Se NC at concentration 500 µg ml−1 displayed outstanding antibacterial activity against E. coli, P. aeruginosa, B. subtilis and S. aureus with inhibition zones 21.26, 20.06, 28.16 and 23.1 mm respectively. From these data, MnO2–Se NC gave highest efficacy toward B. subtilis, while the lowest efficacy was against P. aeruginosa.

Fig. 5. Antibacterial activity of the prepared MnO2–Se NC and other start metals using agar well diffusion method. Letters a, b, c, … mean power significance.

Fig. 5

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The antibacterial potency of the synthesized MnO2–Se NC was quantitatively evaluated by determining MIC, MBC, and the MBC/MIC ratio against all tested bacterial strains (Table 2). These parameters provide essential insight into both the inhibitory and bactericidal behavior of antimicrobial agents.65 The obtained MIC values demonstrated strong antibacterial activity of MnO2–Se NC, with clear variation among bacterial species. The lowest MIC value was observed against B. subtilis (3.9 µg ml−1), indicating that this Gram-positive bacterium is highly sensitive to the nanocomposite. This high susceptibility may be attributed to the absence of an outer membrane in Gram-positive bacteria, allowing easier penetration of nanoparticles and direct interaction with the cytoplasmic membrane.66 The relatively small MIC value suggests efficient disruption of essential cellular processes even at low nanocomposite concentrations. The corresponding MBC value (15.62 µg ml−1) confirms that slightly higher concentrations are sufficient to achieve complete bacterial killing, reflecting strong bactericidal efficiency. S. aureus exhibited moderate sensitivity, with MIC and MBC values of 7.81 and 31.25 µg ml−1, respectively. The increased concentration required compared with B. subtilis may be related to differences in cell wall composition, metabolic adaptability, and stress-response mechanisms.67 The MBC/MIC ratio of 3 demonstrates that MnO2–Se NC does not merely suppress bacterial growth but induces irreversible ce]llular damage leading to cell death.68 Among Gram-negative bacteria, Escherichia coli showed an MIC value of 15.62 µg ml−1 and an MBC value of 31.25 µg ml−1, yielding an MBC/MIC ratio of 2. The higher MIC compared with Gram-positive strains is expected due to the presence of an outer lipopolysaccharide (LPS) membrane that acts as an additional permeability barrier against nanoparticles.69 Despite this structural defense, the relatively low MBC/MIC ratio indicates rapid bactericidal action once inhibitory concentration is reached. This finding suggests that MnO2–Se NC effectively overcomes membrane protection, possibly through oxidative stress and membrane destabilization. Pseudomonas aeruginosa displayed the highest resistance among tested bacteria, with MIC and MBC values of 31.25 and 125 µg ml−1, respectively, and an MBC/MIC ratio of 4. This organism is widely recognized for intrinsic antimicrobial resistance due to low membrane permeability, active efflux pumps, and strong adaptive stress responses.70 The higher concentrations required for inhibition and killing reflect these defense mechanisms.71 However, the ability of MnO2–Se NC to achieve bactericidal activity even against this highly resistant pathogen highlights its strong antimicrobial potential. The MBC/MIC ratios ranging from 2 to 4 across all bacterial strains are particularly significant, as values ≤4 are generally interpreted as indicative of bactericidal activity.72 This confirms that MnO2–Se NC primarily kills bacterial cells rather than only inhibiting growth.

Table 2. MIC, MBC and MBC/MIC ration of MnO2–Se NC toward all tested bacterial strains.
Bacterial strain MIC (µg ml−1) MBC (µg ml−1) MBC/MIC
E. coli 15.62 31.25 2
P. aeruginosa 31.25 125 4
B. subtilis 3.9 15.62 3
S. aureus 7.81 31.25 3
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The remarkable antibacterial performance of MnO2–Se NC can be explained by multiple synergistic mechanisms. The MnO2 surface may catalyze redox reactions that generate reactive oxygen species (ROS), while selenium contributes to oxidative stress and disruption of microbial metabolic pathways. These processes collectively lead to membrane damage, increased permeability, leakage of intracellular constituents, enzyme inactivation, and DNA damage.73,74 The multi-target mode of action likely explains the strong bactericidal behavior observed and suggests a reduced probability for rapid resistance development compared with conventional antibiotics.

Antifungal activity

The antifungal activity of the synthesized MnO2–Se NC was evaluated as illustrated in Fig. 6 and Table 3. The results demonstrated that MnO2–Se NC (500 µg ml−1) exhibited antifungal efficacy compared with the precursor compounds, confirming that nanoscale integration of manganese and selenium significantly enhances antifungal performance. The inhibition zone diameters indicated that C. albicans was more susceptible to MnO2–Se NC than A. brasiliensis, suggesting species-dependent antifungal sensitivity. This difference was further validated by MIC and MFC measurements, where C. albicans showed lower MIC (31.25 µg ml−1) and MFC (62.5 µg ml−1) values compared with A. brasiliensis (MIC = 125 µg ml; MFC = 500 µg ml−1). The calculated MFC/MIC ratios of 2 and 4 indicate that MnO2–Se NC exhibits fungicidal activity rather than merely inhibiting fungal growth.

Fig. 6. Antifungal activity of the prepared MnO2–Se NC and other start metals using agar well diffusion method. Letters a, b, c, … mean power significance.

Fig. 6

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Table 3. MIC, MFC and MFC/MIC ration of MnO2–Se NC toward all tested fungal strains.
Fungal strain MIC (µg ml−1) MFC (µg ml−1) MFC/MIC
C. Albicans 31.25 62.5 2
A. brasiliensis 125 500 4
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The greater susceptibility observed for C. albicans may be related to differences in cell-wall composition, membrane characteristics, and other species-specific physiological properties. Previous studies have suggested that interactions between nanomaterials and fungal cell surfaces may affect membrane integrity and cellular homeostasis.75–77 Once in contact with the fungal surface, nanoparticles adhere electrostatically to negatively charged cell wall components, promoting nanoparticle accumulation and localized damage. This interaction increases membrane permeability, leading to leakage of intracellular biomolecules such as proteins, nucleic acids, and ions, ultimately resulting in loss of cellular homeostasis.77

In contrast, the lower susceptibility of A. brasiliensis may be associated with its filamentous growth form and the structural characteristics of its cell wall, although these factors were not directly investigated in the present study.78,79 These protective features explain the higher MIC and MFC values observed for A. brasiliensis and are consistent with the known resistance of filamentous fungi to nanomaterial-based antifungal agents.

The antifungal mechanism of MnO2–Se NC is likely multifactorial and involves oxidative stress–mediated toxicity. MnO2 nanoparticles can catalyze redox reactions that generate reactive oxygen species (ROS), including hydroxyl radicals and superoxide ions.77 Simultaneously, selenium nanoparticles are known to disrupt cellular redox balance by interfering with antioxidant enzymes such as catalase and glutathione-related systems. Excessive ROS accumulation induces lipid peroxidation in fungal membranes, protein oxidation, mitochondrial dysfunction, and DNA damage. These oxidative events trigger apoptosis-like programmed cell death pathways frequently reported in fungal cells exposed to nanomaterials.80 The observed fungicidal activity (MFC/MIC ≤ 4) suggests that MnO2–Se NC causes irreversible cellular damage rather than temporary metabolic suppression. Furthermore, nanoparticle interaction with fungal spores may inhibit germination and hyphal elongation, thereby preventing colony establishment and fungal propagation.81

The enhanced antifungal efficiency of the MnO2–Se nanocomposite compared with individual metal salts highlights the importance of synergistic nanoscale effects. The high surface area of the nanocomposite increases contact probability with fungal cells, while the combined redox activity of MnO2 and selenium promotes sustained oxidative stress.71 Thus, the results demonstrate that MnO2–Se NC possesses strong broad-spectrum antifungal activity, with particularly high efficacy against yeast pathogens and moderate but significant activity against filamentous fungi. The fungicidal nature, multi-target mechanisms, and enhanced activity relative to precursor compounds suggest that MnO2–Se NC represents a promising candidate for antifungal applications in biomedical, pharmaceutical, and environmental antimicrobial strategies.

Larvicidal and adulticidal activity against C. pipiens mosquitos

Different concentrations of WPW extract (100, 200, 300, 400, and 500 ppm) and MnO2–Se NC (10, 20, 30, 40, and 50 ppm) were administered to C. pipiens mosquito larvae in their third instar. Based on the results displayed in Table 3, WPW extract's the LC50 and LC90 values were found to be 65.996 ppm and 82.455 ppm, respectively, whereas MnO2–Se NCs showed the lowest values, with 5.621 ppm for LC50 and 6.875 ppm for LC90. At 500 ppm for WPW extract and 50 ppm for the other extracts, the larvicidal activity of WPW extract and MnO2–Se NCs peaked. The percentage of larval mortality attained was 100% for Mn–Se NPs and 91.67% for WPW extract. No larval death was observed in the groups treated with sodium selenite and manganese acetate alone, without the WPW extract (Table 4). When LC50 and LC90 values of the WPW extract were recorded at 82.358 and 100.509 ppm, respectively, and the MnO2–Se NCs were measured at 5.663 and 7.002 ppm, it became evident that the same concentration that had previously been treated with larvae exhibited high adulticidal activities against adult C. pipiens. At 500 ppm of WPW extract, adult mortality was 93.33%, and at 50 ppm of MnO2–Se NCs, it was 96.67% (Table 5). In general, it was discovered that adults of C. pipiens were more vulnerable to nanocomposite than WPW extract one. In many countries, especially developing ones, mosquito-borne illnesses are associated with significant public health risks. The growing tolerance of mosquitoes to conventional pesticides has accelerated the development of green nanomaterials as environmentally benign alternatives for mosquito control.82 Extracts from plants are thought to be efficient pesticides.83 Current studies highlight the potential of green-fabricated mosquitocidal nanoparticles as an environmentally friendly replacement for just the plant extract under investigation may yield better outcomes.83 According to recent research, bimetallic nanoparticles are more biologically effective than monometallic nanoparticles at controlling mosquitoes.84 Extensive research has demonstrated the potential of green-fabricated nanoparticles as effective larvicidal agents against Cx. pipiens, offering a sustainable alternative to conventional chemical insecticides.24,25,85 We hypothesized that the green biosynthetic nanoparticles' toxicity to mosquitoes would be caused by their small size, which promotes their entry through the insect cuticle and enter individual cells. Once inside, they interfere with biological processes such as molting. According to the current research, the WPW extract and MnO2–Se NCs larvicidal activities on Cx. pipiens mosquitoes demonstrated toxicity. The MnO2–Se NC had LC50 and LC90 values of 5.621 ppm and 6.875 ppm, respectively, whereas the WPW extract had LC50 and LC90 values of 65.996 ppm and 82.455 ppm. It was discovered that the greatest larvicidal activity of WPW extract and MnO2–Se NCs occurred at 500 ppm for WPW extract and 50 ppm for nanocomposite. For WPW extract and MnO2–Se NCs, the larval mortality percentages were 91.67 and 100, respectively. The groups treated with sodium selenite and manganese acetate just, without the WPW extract, did not exhibit any larval mortality. Plant extracts are used to create manganese or selenium nanoparticles, which have potent biological property and adsorptive capacity. This is explained by the interaction of NH, C Created by potrace 1.16, written by Peter Selinger 2001-2019 O, COO–, and C–N functional groups of proteins with manganese or selenium nanoparticles at the nanoscale.86 Numerous investigations have demonstrated the possibility for using plant or marine-derived nanoparticles as mosquito control agents.85 Biosimilars must be used to control mosquito vectors because Mn and Se nanoparticles have been found to be potential alternatives to chemical synthetic pesticides.87 Similarly, plant-derived extracts have been extensively employed in the green synthesis of nanoparticles with pesticidal properties. For instance, Lagenaria siceraria-mediated ZnO nanoparticles exhibited remarkable larvicidal activity against Anopheles stephensi, with an LC50 value of 56.46 ppm.88 Compared to S. officinalis crude extract, the larvicidal efficacy demonstrated by So-ZnO-NPs against the studied mosquitoes was significantly stronger.25 In a comparative study, Minal and Prakash89 demonstrated that bimetallic nanoparticles exhibited superior larvicidal activity against third-instar Anopheles larvae compared with monometallic nanoparticles. Additionally, our results showed that adult C. pipiens mosquitoes were strongly adulticidal. The WPW extract's reported LC50 and LC90 values were 82.358 and 100.509 ppm, respectively, whereas the MnO2–Se NC were 5.663 and 7.002 ppm. At 500 ppm of WPW extract, the adult mortality rate was 93.33%, but at 50 ppm of Mn–Se NPs, it was 96.67%. Compared to crude extract, adults of C. pipiens generally showed increased susceptibility to nanoparticle extracts, especially those that included two distinct metals. These results are in agreement with the findings of Soni, Prakash,90 who observed enhanced adulticidal efficacy of silver nanoparticles synthesized using Azadirachta indica against Culex quinquefasciatus. Following four hours of exposure, the LC50 value was determined to be 1.06 µL cm−2. Our current study's results are consistent with those of Benelli,91 discovered that AgNPs derived from Acacia caesia leaf extract exhibited a potent adulticidal effect on An. Ae. Albopictus (LD50 = 20.94 µg ml−1), C. tritaeniorhynchus (LD50 = 22.63 µg ml−1), and Subpictus (LD50 = 18.66 µg ml−1). The findings of Suresh et al.92 demonstrated that the crude extract exerted effective adulticidal activity, as reflected by LC50 and LC90 values of 174.14 and 422.29, respectively. Silver nanoparticles biosynthesized from Phyllanthus niruri showed LC50 and LC90 values of 6.68 and 23.58 ppm, respectively. Exposure to these treatments caused adult mortality. When it comes to controlling mosquitoes, bimetallic nanoparticles work better than monometallic ones. The coexistence of two distinct metal interfaces contributes to enhanced biological performance, leading to superior larvicidal and adulticidal activities compared with monometallic counterparts.26,93

Table 4. Larvicidal activity of the WPW extract and MnO2–Se NC for the mosquito vector, Culex pipiensa.

Treatments Concentrations (ppm) Larval mortality % ± SD LC50 (ppm) LC90 (ppm)
WPW extract Control 0.0 ± 0.0a 65.996 82.455
100 13.33 ± 0.577b
200 33.33 ± 0.577c
300 56.67 ± 1.155d
400 78.33 ± 0.577e
500 91.67 ± 0.577f
MnO2–Se NC Control 0.0 ± 0.0a 5.621 6.875
10 26.67 ± 0.577b
20 56.67 ± 1.155c
30 73.33 ± 0.577d
40 91.67 ± 0.577e
50 100.00 ± 0.00e
Mn (CH3COO)2·4H2O Nil Nil Nil Nil
NazSeO3 Nil Nil Nil Nil
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a

Letters a, b, c, … mean power significance.

Table 5. Adulticidal activity of WPW extract and MnO2–Se NC for the mosquito vector, Culex pipiensa.

Treatments Concentrations (ppm) Larval mortality % ± SD LC50 (ppm) LC90 (ppm)
WPW extract Control 0.0 ± 0.0a 82.358 100.509
100 11.67 ± 0.577b
200 36.67 ± 0.577c
300 58.33 ± 0.577d
400 73.33 ± 0.577e
500 93.33 ± 0.577f
MnO2–Se NC Control 0.0 ± 0.0a 5.663 7.002
10 23.33 ± 1.155b
20 53.33 ± 1.528c
30 71.67 ± 1.155d
40 88.33 ± 1.155e
50 96.67 ± 0.577f
Mn (CH3COO)2·4H2O Nil Nil Nil Nil
NazSeO3 Nil Nil Nil Nil
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a

Letters a, b, c, … mean power significance.

Conclusion

The present study successfully demonstrated the green synthesis of MnO2–Se NC using watermelon peel waste as a natural reducing and stabilizing source. UV-Vis, FTIR, XRD, TEM, and DLS analyses confirmed the formation of predominantly spherical nanostructures with an average size of approximately 55 nm. The synthesized MnO2–Se NC exhibited strong antibacterial activity, with MIC values of 3.9–31.25 µg ml−1 and bactericidal effects, particularly against Bacillus subtilis, as well as notable antifungal activity, with greater efficacy against Candida albicans than Aspergillus brasiliensis. The nanocomposite also showed pronounced larvicidal and adulticidal activity against Culex pipiens at substantially lower concentrations than the crude watermelon peel extract. These findings demonstrate that watermelon peel waste can be effectively valorized for the production of a nanocomposite with promising in vitro antimicrobial and laboratory-based mosquitocidal activities. However, the study was limited to laboratory evaluations, and the mechanisms of action, toxicity, environmental fate, long-term stability, and effects on non-target organisms were not investigated. Future studies should therefore address these aspects, together with synthesis scalability, before the practical applicability of MnO2–Se NC can be established.

Author contributions

Conceptualization, S. S., T. A. S., A. H. H., and E. S.; methodology, S. S., A. S. A., T. A. S., A. H. H., M. H. A., H. S. G., A. A. K., N. S. B. and E. S.; writing – original draft preparation, T. A. S., A. H. H., and E. S.; writing – review and editing, S. S., A. S. A., T. A. S., A. H. H., M. H. A., H. S. G., A. A. K., N. S. B. and E. S. All authors have read and agreed to the published version of the manuscript.

Conflicts of interest

The authors declare that they have no conflicts of interest.

Acknowledgments

This work was supported and funded by Princess Nourah bint Abdulrahman University Researchers Supporting Project number (PNURSP2026R357), Princess Nourah bint Abdulrahman University, Riyadh, Saudi Arabia.

Data availability

All data analyzed during this study are available within the current work.

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Data Availability Statement

All data analyzed during this study are available within the current work.

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