Antibacterial Application of Heterogeneous CuO-NiO-ZnO Metal Oxides Nanocomposites

Antibacterial Application of Heterogeneous CuO-NiO-ZnO Metal Oxides Nanocomposites | 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 Research Article Antibacterial Application of Heterogeneous CuO-NiO-ZnO Metal Oxides Nanocomposites Vikas Choudhary, Kusham Lata, Manish Kumar, Ajay Sharma, Raman Kumar, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7532947/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract The synthesis, characterization, & use of mixed metal oxides, a unique family of compounds, have attracted ever-increasing interest as they offer an excellent opportunity to refine the desired properties for improved functional performance in interaction with base metal oxides. In this study, NiO-ZnO-CuO composites are synthesized by using simple co-precipitation technique at 200 o C and 500 o C temperature and then characterized for its properties by UV-visible, X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) studies, Dynamic Light Scattering (DLS) and Scanning electron microscopy (SEM) analysis. At 200°C & 500°C, the CuO-NiO-ZnO nanocomposite had an optical band gap energy of 2.70 eV & 2.67 eV, respectively. The UV-visible spectroscopy was utilized to find the band gap using the Tauc plot. XRD studies indicated the presence of well-defined CuO (monoclinic), NiO (cubic), & ZnO (hexagonal) phases, with the maximum fraction of volume belonging to the NiO. The images of SEM by mixed oxide nanocomposites display nanoparticles in the form of irregular shapes. The phases indicate the presence of trimetallic oxide heterojunctions, which strongly impact the overall properties of nanocomposites (NCs). Also, the antibacterial activities of nanocomposites are studied, which showed that at 200°C, the nanocomposite exhibited antibacterial action against gram-negative bacteria (GNB), whereas at 500°C, it demonstrated efficacy against gram-positive bacteria (GPB) and also affected GPB at higher doses. CuO-NiO-ZnO nanocomposite antibacterial concentration Figures Figure 1 Figure 2 Figure 3 Figure 4 Highlights Synthesis of CuO-NiO-ZnO nanocomposites by the co-precipitation method. Characterization techniques used for the assurance of CuO-NiO-ZnO nanocomposites formation were UV-visible, FTIR, XRD, DLS, and SEM analysis. Enhanced antibacterial efficacy of CuO-NiO-ZnO against Bacillus , Pseudomonas & E. coli bacterial strains. 1. Introduction The scientific fields of engineering, chemistry, physics, and materials science have experienced an increased interest in the development of applications and research on mixed metal oxides. Combining multiple metals together in the oxide framework can develop materials having unique chemical & physical characteristics that result in comparatively better performance in wide range of applications. The metals may exhibit the characteristics of isolated entities that contribute their qualities to the system, or they may have altered properties by interactions between metals or between metals and oxygen [ 1 ]. A nanostructured system's performance can be significantly enhanced by the interaction of its many constituent materials. At the microscopic level, the close contact surfaces of composite materials with corresponding band potentials improve the efficiency of charge separation, lengthen the life of charge carriers, & improve transfer of charge [ 2 ]. Most of the mixed oxides (MO) are single-phase materials that can reveal functional properties resulting from combining individual oxide functionalities. Multiphase nanocomposites, created by the coexistence of two MO within the same surrounding substance, have been made for applications such as photocatalysis & antibacterial application [ 3 ]. Various metal-oxide nanocomposites, including MnOx- CeO₂ [ 3 ], ZnO- CuO [ 4 ], NiO- ZnO [ 5 ], and MgO- ZnO [ 6 ], were synthesized and studied using a variety of methodologies. Amongst these, the widely utilized composite MnOx supported on Ti 6 Al 4 V has been synthesized, which has influenced its properties. According to these studies, composite materials trap photogenerated carriers more effectively than monomeric oxides. A few studies have shown enhanced device performance and characteristics for MO NCs with three or more phases, such as ZnO-WO₃-ZnWO₃ [ 2 ], Mo-W-V MOs [ 7 ], La-V-Mo oxide solid solution [ 8 ], and the pseudo-quaternary system CaO-SiO₂-CoO [ 9 ]. Some conclusions have been reported by another investigation on "ZnO-CeO 2 -TiO₂" composites made by combustion, which revealed improved photocatalytic color removal and outstanding UV absorption. [ 10 ] Infectious disorders caused by microorganisms like fungi, viruses, parasites, or bacteria have recently affected public health in many countries and are one of the leading causes of mortality worldwide. Because of their electron-donating qualities, metallic oxide-composited components are known to encourage the production of the activated oxygen class, according to researchers. Researchers have recently discovered the antibacterial properties of different metal oxide NCs [ 11 ]. There have been reports of nanocomposites targeted at both Gram-positive and Gram-negative microorganisms. The targeting of the bacteria was identified through an electrostatic interaction between the positive charge residues on the composite surface and the negative charge microbial membrane. Copper oxides NPs have attracted significant attention among other metal oxide nanoparticles. CuO is a low-bandgap (1.2 eV) p-type semiconductor that finds utility as a high-Tc superconductor in sensing gas, photocatalysis, photovoltaics, magnetic storage, & technology. By preventing the development of fungi, viruses, bacteria, & algae, CuO NPs in particular exhibit strong antibacterial qualities. When added to coatings, copper oxide nanoparticles (CuO NPs) can function as antifungal and antibacterial agents. [ 12 ]. In recent years, nickel oxide (NiO) which is a p-type semiconductor with a broadband gap of 3.8eV (3.6–4.0 eV), has garnered a lot of interest because of its uses in gas sensoring application, pigments, spin valve devices, alkaline battery cathodes, etc. [ 13 ] NiO is regarded as an appropriate material for solar cells, antiferromagnetic layers, lithium-ion battery anodes, and other applications because of its high theoretical capacity, quick reaction kinetics, & strong cyclic reversibility [ 14 ]. NiO is extremely sensitive to a wide range of gases, including ethanol, CO, NO 2 , ammonia, and acetone [ 15 ]. High chemical stability, exceptional capacity of electron transfer, & anti-inflammatory qualities are all possessed by NiO nanoparticles. Their special qualities, such as release of metal ion, surface area, & adsorbing capacity, also provide them with cytotoxic effects [ 16 ]. The studies on zinc oxide nanostructures are gaining attentions because of their distinctive characteristics and numerous uses. Using ZnO nanoparticles has the benefit of significantly reducing the activity of harmful bacteria at low concentrations. Since ZnO nanoparticles have antifungal & antibacterial properties at lower concentrations, their thin coatings can be utilized for developing products that are resistant to bacteria. ZnO is a member of the metal oxide class, which is distinguished by its ability to photocatalyze and photooxidize biological and chemical species. It makes use of a multipurpose nano platform that releases reactive oxygen species (ROS) to attack cancerous cells from the outside [ 17 ]. ZnO nanoparticles' antibacterial effects result from an electrical interaction between the greater cell damage as a result of the nanoparticles' increased interaction with the cell surface. Previous studies showed that the wavelength-dependent fluorescence of excitation by CuO.La₂O₂Co₃.ZnO [28], Co₃O₄.CeO₂.ZnO [29], and La₂O₂Co₃.CeO₂.ZnO [27] nanocomposites has been studied by scientists. When exposed to methyl violet 6b (MV), the La₂O₂Co₃.CeO₂.ZnO composite exhibits the 93.75% efficiency of photocatalytic degradation. For the same dye, the NiO.CeO₂.ZnO [30] composite shows 96.07% efficiency. It also demonstrates powerful antimicrobial activity against harmful bacteria such as P. mirabilis, with a zone of inhibition (ZOI) of 15 mm. Ag₂O, CeO₂, and ZnO exhibit antibacterial action against K. pneumoniae with a ZOI of 15 mm & a photocatalytic destruction rate of 84.04% against MV. Scientists have reported the synthesis & characterization of CdO–ZnO–NiO mixed MO nanocomposite for the photocatalytic & antibacterial activities of the microwave-assisted nanocomposite [31]. The combination of these three distinct metallic-oxide phases forms a heterogeneously mixed metal oxide “CuO-NiO-ZnO nanocomposite” that provides a range of options for controlling the nanocomposite's characteristics & defining novel technological applications in the antibacterial activity for targeting a wide variety of hazardous microbes. 2. Experimental Details 2.1 Materials Materials and methods All chemicals used were of analytical grade and used without further purification. Nickel nitrate hexahydrate (Ni (NO 3 ) 2 .6H 2 O), Copper Sulphate Pentahydrate (CuSO4.5H2O), zinc chloride (ZnCl2), Sodium Hydroxide was purchased from LOBA CHEMIE PVT, Ethanol. Bacterial strains were initially procured from the Institute of Microbial Technology in Chandigarh, India. 2.2 Synthesis of the Nanocomposites CuO-NiO-ZnO nanocomposite was synthesized via the co-precipitation method. A 0.01 M predecessor solution of three salts was prepared by weighing 300 mg of Cu, 400 mg of Ni, and 500 mg of Zn salts, and then mixing it with the 50 ml solution containing 40 ml of water with 10 ml of ethanol. The solution was swirled magnetically at temperature of 50°C for time period of 30 minutes. To make a 50 ml mixed solvent of NaOH, two grams of NaOH pellets were dissolved in five minutes of distilled water. The solution of NaOH is then added to the combined precursor solution with continuously stirring of solution. A precipitate is seen to form immediately after the color of mixture changes from bluish-green to a shade of blackish-green. After five minutes, the temperature is raised to 100℃ & maintained there for an additional half-hour while being continuously agitated. The mixture was then let to cool at room temperature. Filtration method was utilized to collect the precipitate, which was then washed three times with distilled water & dried on a hot plate at 100℃ for two hours. A fine black powder is then produced by annealing the powder for two hours at 200 and 500 ℃ [ 1 ]. 2.3 Characterizations The properties of the synthesized materials (CuO-NiO-ZnO nanocomposite) were thoroughly investigated using various characterization techniques. Fourier Transform Infrared (FTIR) spectroscopy (Perkin Elmer, subtech spectrum) was employed to classify the functional groups within CuO-NiO-ZnO nanocomposite in the 4000 − 400 cm − 1 spectral range. Powder X-ray Diffraction (XRD) analysis using a PAN analytical EMPEREAN diffractometer with Cu K alpha radiation (λ = 1.5405980 Å) provided insights into the crystallinity, phase composition, and crystallite size of the materials. Field Emission Scanning Electron Microscopy (FESEM) & Energy Dispersive Spectroscopy (EDS) were utilized to investigate the morphology, size, and surface topography, while EDS analysis provided composition data. UV-visible spectroscopy (Specord 200 PLUS) was employed to determine optical properties, including band gap estimation. Dynamic light scattering (DLS) is a quasi-elastic light dispersion technique which is used to find out the size distribution of selective NPs and is also used for evaluation of the size & dispersion of the nanocomposite. 2.4 Determination of antibacterial activity The antibacterial property of CuO-ZnO-NiO nanocomposite was evaluated using the agar well diffusion method against two GNB, Escherichia coli (MTCC 1652), & Pseudomonas aeruginosa (MTCC 741), as well as one GPB, Bacillus subtilis (MTCC 121). The Institute of Microbial Technology in Chandigarh, India, was the original source of the bacterial strains. Overnight cultures of the bacteria were diluted to approximately 10 8 CFU/mL using sterile distilled water, & 1 mL of the diluted inoculum was spread onto nutrient agar plates (25 mL per plate). Six to eight wells (7 to 8 mm in diameter) were punched into the agar using a sterilized cork borer. The wells were then filled with 1 mL of freshly prepared CuO-NiO-ZnO nanocomposite solutions at concentrations ranging between 0.05 µL to 5 µL for the first screening, and 1 mg/mL for the subsequent evaluation. Dimethyl sulfoxide (DMSO) was used as a solvent for the test compounds and also functioned as a negative control. Plates are incubated at a temperature of 37°C for a time period of 24 & 48 hours, and the zone of inhibition is measured in millimeters to assess the antimicrobial activity. Bacitracin & Chloramphenicol (10 mg/mL) were used as positive controls. In order to assess the antibacterial potential of the nanocomposites, the inhibitory zones surrounding the wells were inspected and documented after the incubation period. The modified agar well diffusion method is utilized to test the minimum inhibitory concentration (MIC) of each compound, giving an inhibitory zone at a concentration of 1 mg/mL. Several wells in the agar plates received various applications of a single chemical at different concentrations (1,000–1 µg/mL). Each dilution was added to wells at a volume of about 1 mL. During 48 hours, all test plates were incubated at 37°C. A definite zone of inhibition was visible in the lowest concentration of each chemical, which was referred to as the MIC. 3. Results and discussion 3.1 UV–visible analysis: UV-visible absorption spectra & Tauc plots of the synthesized mixed MO nanocomposites at 200 and 500 ℃ as represented in Fig. 1 (a & b). The UV region, which ranges from 200 to 400 nm, is where the distinctive peaks in the Ultraviolet-Visible analysis can be observed. The graph shows a prominent SPR absorbance peak at a specific wavelength (λmax) of 288nm & 296nm, confirming the synthesis of CuO NPs upon reduction [ 18 ]. On the other hand, it shows an absorbance peak at a specific wavelength (λmax) of 338nm, indicating the presence of NiO [19]. In the case of ZnO absorbance peak at a specific wavelength (λmax) of 360nm & 372nm [20]. The bandgap of the CuO-ZnO-NiO nanocomposites, which are annealed at a temperature of 200 ℃, was determined to be 2.70 eV with a slight change to 2.67 eV following annealing at 500°C. 3.2 Dynamic Light Scattering (DLS): Dynamic light scattering (DLS) is a quasi-elastic light dispersion technique that is used to evaluate the dispersion & size of NPs as well as to determine the size distribution of selected NPs. [63]. The particle size distribution of the mixed metal oxides of CuO/NiO/ZnO NCs solution, which is annealed at 200°C, is shown in Fig. 1 (c). Depending upon the results of DLS, 100% of the particles have a diameter of 450.9 nm. In addition, the DLS provides results shown in Fig. 1 (d) that 79.0% of the particles have a diameter of 717.1 nm, 182.8 nm by 19.3% of the particles, and 5118 nm by 1.7% of the particles in mixed metal oxides of CuO/NiO/ZnO NCs solution, which is annealed at 500°C. The Poly-dispersity Index (PdI) for mixed metal oxides of CuO/NiO/ZnO NCs solution, which is annealed at 200°C & 500°C, are 0.280 & 0.352, respectively. It represents that both the mixed metal oxides of CuO/NiO/ZnO NCs solution, which are annealed at 200°C & 500°C, respectively, have a particle distribution of 'mono-scattered' order that explains the distribution of particle size, which consists of a one-size mode without an aggregate mode. The PDI having values between 0.1 to 0.7 (both values included) are mostly monodispersive in nature, While PDI value greater than 0.7 causes the broadening of size distribution of macromolecular system in solution & non-monomodal distribution approaches for data analyses should be considered [64]. 3.3 XRD Analysis: X-ray diffraction (XRD) spectrum was obtained to investigate the crystallographic structure of the synthesized CuO-NiO-ZnO nanocomposite annealed at 200°C & 500°C in Fig. 2 (a). Annealing MO composite [CuO-NiO-ZnO] nanoparticles at temperatures of 200 & 500 ℃ changes their XRD properties. Only the peaks corresponding to Zn-O & Cu-O were identifiable after heating to 200°C. Several of the diffraction peaks of the different oxides overlap due to their very similar angles of diffraction. On the other hand, there was no peak of diffraction that was linked to the NiO phase. Because some of the byproducts of the precursors may still be present, Ni (OH) 2 may persist at a temperature of 200°C. Research on NiO-ZnO MO nanoparticles, which were produced in a way similar to that of CuO-ZnO-NiO samples, revealed the existence of ZnO & β-Ni (OH)₂ mixed phases upon annealing at a temperature of 200°C. Previous studies using TG-DSC, XRD, and HRTEM examinations have shown that, depending on the manufacturing procedure, Cu(OH)₂ & Zn(OH)₂ transform into their corresponding oxides at temperatures below 150°C [25] and slightly over 300°C [26], respectively. After being annealed at a temperature of 500°C, the peaks of diffraction for NiO, ZnO, & CuO have been clearly visible, indicating that the remaining hydroxides have fully broken down and pure metal oxides have formed. The end product is a nanocomposite that has a combination of several binary oxide phases. Table 1 XRD peaks and the corresponding planes obtained from CuO-NiO-ZnO nanocomposites annealed at 200°C & 500°C. Samples 2θ (degree) d-spacing (nm) Plane (s) CuO-NiO-ZnO (200°C) 31.84 2.808 100 32.81 2.727 110 34.47 35.65 36.23 38.98 47.73 54.4 56.68 59.02 62.09 62.85 68.16 2.599 2.516 2.477 2.308 1.903 1.685 1.622 1.563 1.493 1.477 1.374 002 111 101 200 102 020 110 202 -113 103 112 CuO-NiO-ZnO (500°C) 31.83 34.6 35.74 36.40 37.14 38.92 43.24 47.74 48.88 56.71 62.81 66.35 68.17 69.09 2.809 2.590 2.510 2.466 2.418 2.312 2.090 1.903 1.861 1.621 1.470 1.407 1.358 1.358 100 002 111 101 111 200 200 102 -202 110 220 022 112 201 3.4 SEM analysis: Scanning electron micrographs of the CuO-NiO-ZnO mixed oxide nanocomposites at different magnifications for the samples prepared are presented in Fig. 2 (b) & (d), annealing at 200°C and 500°C, respectively. The observed shape is spherically irregular & the size distribution observed to be heterogeneous, having some cavities. The morphology of the nanocomposites is observed to vary depending upon the concentration of precursor solution, temperature & many other factors. The average size of particles of nanocomposites at different temperatures (200°C and 500°C) was observed as 49.64nm and 28.96nm, respectively [Fig. 2 (c) & (e)]. The sizes of particles calculated in SEM & XRD have some slight variation. The reason for this diminutive difference is that calculations by SEM were based upon the difference between the visible boundaries of grains; on the other hand, calculations of XRD have measured the extended crystalline region, which coherently diffracts X-rays. The image of SEM has shown the aggregated particle, which is due to the excess heat generated during the process of calcination. The size of particles has some minimum changes that occur due to the aggregation of particle. 3.5 Fourier Transform Infrared Spectroscopy (FTIR): FTIR spectroscopy gives important information on the functional groups and chemical interactions inside the synthesized CuO-NiO-ZnO nanocomposites in Fig. 2 (f). In the FTIR spectra of synthesized CuO-NiO-ZnO nanocomposites, the distinct characteristic peaks were observed. The absorption bands below 600 cm − 1 are mostly linked to the CuO-NiO-ZnO group. Metal-oxygen bond vibrations often exhibit FTIR absorption peaks below 1000 cm − 1 . CuO has exhibited six infrared vibration bands at 147, 161, 321, 478, 530, & 590 cm − 1 [21]. The Zn-O bond vibrational bands may be found at approximately 395, 425, 470, & 515 cm − 1 , depending upon the structure & shape of the nanoparticles [22, 23]. The IR absorption peaks in NiO are caused by NiO vibrations at 454 cm − 1 & 571 cm − 1 [24]. It is clear that the vibration peaks of Ni-O, Zn-O, & Cu-O that appear in the region below 600 cm − 1 overlap. Table 2 FTIR peaks with the functional groups of the synthesized CuO-NiO-ZnO NCs annealed at 200 º C & 500ºC. Compounds Peak position (cm − 1 ) Nature Type of Molecular motion CuO-NiO-ZnO (200°C) 3462 cm − 1 Strong broad O-H stretching 2286 cm − 1 Weak broad C = N stretching 2048 cm − 1 Medium C = C = C stretching 1983 cm − 1 1723 cm − 1 1351 cm − 1 1140 cm − 1 Medium Strong Medium Strong C = C = C stretching C = O stretching O-H Bending C-O stretching CuO-NiO-ZnO (500°C) 1665 cm − 1 Strong broad C = O stretching 1620 cm − 1 Strong C = C stretching 1430 cm − 1 medium O-H stretching 1142 cm − 1 medium C-O stretching 4. Antimicrobial Application Nanocomposites of tri-metallic nature have shown stronger antibacterial property, as compared to nanoparticles of mono-metallic & bi-metallic nature. The previous study on Au-Pt-Ag nanocomposite synthesized by green method which show effective antibacterial property against the bacteria types, (S. aureus & E. coli) [32–34]. The synthesized CuO/NiO/ ZnO nanocomposite shows antimicrobial action against the GNB ( Pseudomonas & E. coli ) & GPB (Bacillus) . These nanocomposites work against bacteria through different routes. The one of the important route is by reactive oxygen species (ROS) formation. These reactive species then disrupt the cycle of respiratory system, the cycle of protein transfer system, the food metabolism cycle, and DNA replication, causing cell death [35]. Table 3 Tabulation showing ZOI of GPB (Bacillus) & GNB (Pseudomonas & E. coli) with the action of nanocomposites which are annealed at 200 O C. Sr. No Concentration (ppm) ZOI Bacillus (GPB) (mm) ZOI Pseudomonas (GNB) (mm) ZOI E.coli (GNB) (mm) 1. 5 5 2. 10 5 3. 20 12 5 4. 40 19 5 5. 50 20 5 6. 250 20 5 7. 500 20 17 The antibacterial properties of the synthesized mixed MO nanocomposite were examined at several concentrations (5, 10, 20, 40, 50, 250, and 500 ppm). With an increase in concentration, the nanocomposite's antibacterial efficacy against two distinct bacterial strains improved. This could be the result of more reactive species that can stop the development of bacteria [36]. Figure 3 displays pictures of the antibacterial properties of the synthesized mixed MO nanocomposite at different annealing temperatures of 200 & 500°C, respectively. The produced nanocomposite had outstanding antibacterial activity, according to the data. At a dose of 500 ppm, the largest ZOI against the GPB was seen Table 4 Tabulation showing ZOI of GPB (Bacillus) & GNB (Pseudomonas & E. coli) with the action of nanocomposites which are annealed at 500 O C. Sr. No Concentration (ppm) ZOI Bacillus (GPB) (mm) ZOI Pseudomonas (GNB) (mm) ZOI E.coli (GNB) (mm) 1. 5 2. 10 5 3. 20 5 4. 40 9 5. 50 10 6. 250 13 10 7. 500 15 17 Action of Nanocomposites on Bacteria Cell. In addition to being crucial for sustaining bacterial growth and reproduction, typical bacterial metabolic activities can also result in illness. Bacterial cell death results from oxidative stress & damage to the cell membrane of bacterial cells caused by disruptions in bacterial metabolism. The effects of nanomaterials on bacterial metabolism have been explained by a variety of different processes, such as metal ion dissolution and reactive oxygen [60, 61]. Bacterial metabolism is a crucial biofilm function: The development and expansion of S. mutans biofilm depend on d-alanine metabolism [62].The production of ROS, which can damage membranes, DNA, & proteins, directly interacts with the cell membrane due to the ability of certain metal-based Nanocomposites to generate metal ions through dissolution, such as by inhibiting the chain of electron transport, & control of metabolic processes of bacterial cells are some of the ways that Nanocomposites can attack bacterial cells. The attack of Nanocomposites on bacterial cells through numerous mechanisms, as shown in Fig. 4 . The results observed from the comparison between the antibacterial property of the ZnO-NiO-CuO NC examined in this work and previously published data, as observed in Table 5 (a, b, c) below. Table 5 (a). Comparison of antibacterial application by various nanocomposites for gram-positive ( Bacillus ) from the data obtained from earlier reported Studies. Sr. No NCs Method of Synthesis ZOI diameter (mm) References 1 Ni-O green synthesis (Stevia leaf extract) 14 [51] 2 Cu-O green synthesis (Aerva javanica extract) 9 [52] 3 Zn-O green synthesis (Cassia fistula extract) 14 [53] 4 ZnO-CuO green synthesis (Mentha longifolia leaf extract) 15 [54] 5 CdO-NiO-ZnO microwave-assisted method 15 [55] 6 CuO-NiO-ZnO Co-precipitation method 15 present study Table 5 (b). Comparison of Antibacterial Application by various Nanocomposites for Gram-negative bacteria (E. coli) from the Data obtained from earlier Reported Studies. Sr. No NCs Method of Synthesis ZOI diameter (mm) References 1 NiO.CeO 2 . ZnO Co-precipitation method 11 [56] 2 Ag 2 O. CeO 2 . ZnO Co-precipitation method 12 [57] 3 CdS-ZnO Wet chemical synthesis 14 [58] 4 ZnO:Cu/graphene Pyrolysis method 16 [59] 5 CdO−ZnO−NiO Microwave-assisted method 16 [55] 6 CuO−ZnO−NiO Co-precipitation method 17 present study Table 5(c). Comparison of Antibacterial Application by various nanocomposites for Gram-negative bacteria (Pseudomonas) from the data obtained from earlier reported Studies. Sr. No. NCs Method of Synthesis ZOI diameter (mm) References 1 NiO.CeO 2 . ZnO Co-precipitation method 13 [56] 2 Ag 2 O. CeO 2 . ZnO Co-precipitation method 11 [57] 3 CuO−ZnO−NiO Co-precipitation method 20 present study It demonstrates that the produced nanocomposite has stronger antibacterial application as compared to single & bi-metallic nanoparticles that have been previously described. Furthermore, the recently described NCs can be examined for both antioxidant & antibacterial properties, while the CuO-NiO-ZnO NCs, which were previously published, were only examined for antimicrobial activity [37–43]. According to Fig. 3 , the synthesized nanocomposite exhibits superior antibacterial action against Bacillus (GPB) & Pseudomonas & E. coli (GNB). Because of the presence of the thick coating of peptidoglycan in GPB, it is simpler to target & harm the cell wall of the bacterial cell. GNB has an overabundance of lipopolysaccharides in its outer layer. Consequently, strains are more expensive than GPB. Consequently, it is discovered that the combined metal oxide CuO/NiO/ZnO, which is annealed at 500°C, has a stronger inhibitory effect on gram-positive bacteria [44–50]. On the other hand, the combined MO CuO-NiO-ZnO, which is annealed at 200°C, has a stronger inhibitory effect on gram-negative bacteria. GPB & GNB utilize distinct strategies to let the CuO-NiO-ZnO NCs into their cells; this might be due to variations in the composition of their membranes. The composition of the membrane determines how the CuO-NiO-ZnO NCs attach & are transported into the cell membrane of the bacterial cell. 5. Conclusions The chemical synthesis approach is one of the most efficient, economical, and time-saving approaches to developing the CuO-NiO-ZnO NCs. A higher concentration of the CuO-ZnO-NiO NCs proved to enhance its antibacterial activities. This might be because there are more particles accessible to interact with bacterial species. Bacillus (GPB) was more successfully inhibited by synthesized CuO-NiO-ZnO NCs annealed at 500°C than Pseudomonas & E. coli (GNB). On the other hand, Pseudomonas & E. coli (GNB) were more successfully inhibited by the synthesized CuO-NiO-ZnO NCs annealed at 200°C than Bacillus (GPB). The action of CuO-ZnO-NiO NCs upon the two species of bacteria differs because of variations in surface charge and cell wall composition. Using several characterization methods, including SEM, FTIR, XRD, & UV-visible spectroscopy, the physicochemical parameters of the samples were investigated. The study has investigated the application of nanocomposites for the development of antibacterial agents and coatings, which offers a gap of energy by nanocomposites of 2.70 eV & 2.67 eV at different annealing temperatures of 200°C and 500°C, respectively. Sharp & strong peaks observed in the XRD pattern confirmed the extremely crystalline structure of nanocomposites. The SEM micrograph confirms the heavily agglomerated background, which has visible voids at particular points. The study has investigated the application of nanocomposites for the development of antibacterial agents & coatings, which offer innovative strategies to combat infections. Abbreviations MO Metal oxide NCs Nanocomposites NMs Nanomaterials SEM Scanning electron microscopy DLS Dynamic light scattering XRD X-ray diffraction FTIR Fourier-transform infrared spectroscopy UV–Vis Ultraviolet–visible spectroscopy ROS Reactive oxygen species CB Conduction band VB Valence band Declarations Ethics approval and consent to participate This study involved only laboratory bacterial strains; no experiments on humans or animals were performed. According to the journal policies, formal human or animal ethics approval was not required. Informed consent was obtained from all individual participants included in the study. Consent for publication The author confirms that informed consent for publication of the manuscript was obtained from all participants involved in the study. Competing interests The authors declare no competing interests. Authorship contribution Vikas Chaudhary: Writing original draft, experimental, data curation, Kusham Lata- Writing review and editing, investigation, data curation, Manish Kumar: formal analysis, visualization, Ajay Sharma: formal analysis, visualization, Raman Kumar- formal analysis, Vivek Sheel Jaswal- Supervision, conceptualization, writing, review and editing, formal analysis, investigation. Funding This research did not receive any specific grant from funding agencies. Availability of data and materials No datasets were generated or analysed during the current study. Acknowledgements I am Kusham Lata, and I am highly thankful to DST-INSPIRE for providing a funding grant for my research work. We are also grateful to Central University of Himachal Pradesh for providing me with a platform for research work. References J. A. Rodriguez, X. Wang, J. C. Hanson, G. Liu, A. Iglesias-Juez, & M. Fernández-Garci´a. The behavior of mixed-metal oxides: Structural and electronic properties of Ce1−xCaxO2 and Ce1−xCaxO2−x. The Journal of Chemical Physics, (2003) 119(11), 5659–5669. https://doi.org/10.1063/1.1601595. X. Jiang, X. Zhao, L. Duan, H. Shen, H. Liu, T. Hou, and F. Wang. Enhanced photoluminescence and photocatalytic activity of ZnO-ZnWO4 nanocomposites synthesized by a precipitation method. Ceramics International (2016) 42(14) 15160-15165. https://doi.org/10.1016/j.ceramint.2016.05.098. Z. Wang, M. Yang, G. Shen, H. Liu, Y. Chen, and Q. Wang. Catalytic removal of benzene over CeO2–MnO x composite oxides with rod-like morphology supporting PdO. Journal of nanoparticle research (2014) 16(5) 2367. https://doi.org/10.1007/s11051-014-2367-5. R.K. Sharma, R. Ghose. Synthesis of nanocrystalline CuO-ZnO mixed metal oxide powder by a homogeneous precipitation method, Ceram. Int. (2014) 40(7) 10919–10926. https://doi.org/10.1016/j.ceramint.2014.03.089 P.P. Dorneanu, A. Airinei, N. Olaru, M. Homocianu, V. Nica, F. Doroftei, Preparation and characterization of NiO, ZnO and NiO–ZnO composite nanofibers by electrospinning method, Mater. Chem. Phys. (2014) 148(3) 1029–1035. https://doi.org/10.1016/j.matchemphys.2014.09.014 J. Vidic, S. Stankic, F. Haque, D. Ciric, R. Le Goffic, A. Vidy, J. Jupille, B. Delmas, Selective antibacterial effects of mixed ZnMgO nanoparticles, J. Nanoparticle Res. 15(5). 1595. https://doi.org/10.1007/s11051-013-1595-4 G. Schimanke, M. Martin, J. Kunert, H. Vogel, Characterization of Mo-V-W Mixed Oxide Catalysts by ex situ and in situ X-Ray Absorption Spectroscopy, Z. Anorg. Allg. Chem. (2005) 631(6-7) 1289–1296. https://doi.org/10.1002/zaac.200500064 H.R. Arandiyan, M. Parvari, Studies on mixed metal oxides solid solutions as heterogeneous catalysts, Brazilian J. Chem. Eng. (2009) 26 63–74. https://doi.org/10.1590/S0104-66322009000100007 X. Li, R. Zhao, H. Jiang, Y. Zhai, P. Ma, Preparation and Catalytic Properties of ZnOCeO 2 -TiO 2 Composite, Synth. React. Inorganic, Met. Nano-Metal Chem. (2016) 46(5) 775–782. https://doi.org/10.1080/15533174.2014.988806 H. R. Arandiyan, M. Parvari. Studies on Mixed Metal Oxides Solid Solutions as Heterogeneous Catalysts. Braz. J. Chem. Eng. ( 2009) , 26 (1), 63–74. https://doi.org/10.1590/S0104-66322009000100007 C. Gionco, M. C. Paganini, S. Agnoli, A. E. Reeder, E. Giamello. Structural and Spectroscopic Characterization of CeO2–TiO2 Mixed Oxides. J. Mater. Chem. A ( 2013) , 1 (36), 10918. https://doi.org/10.1039/C3TA12018J T. Dodevska, D. Hadzhiev, I. Shterev, and Y. Lazarova. Application of biosynthesized metal nanoparticles in electrochemical sensors. Journal of the Serbian Chemical Society (2022) 87(4) 401-435. https://doi.org/10.2298/JSC200521077D A.K. Srivastava, S. Thota, J. Kumar, Preparation, microstructure and optical absorbtion behavior of NiO thin films, J. Nanosci. Nanotechnol. (2008) 8(8) 4111–4115. https://doi.org/10.1166/jnn.2008.AN36 S.M. Abbas, S.T. Hussain. S. ali, N. Ahmad, N. Ali, S. Abbas, Z. Ali, Modification of carbon nanotubes by CuO-doped NiO nanocomposite for use as an anode material for lithium ion batteries, J. Sol. State. Chem. (2013) 202 43–50. https://doi.org/10.1016/j.jssc.2013.03.036 M. Ben Amor, A. Boukhachem, A. Labidi, K. Boubaker, M. Amlouk, Physical investigations on Cd doped NiO thin films along with ethanol sensing at relatively low temperature, J. Alloys Compnd. (2017) 693 490–499. https://doi.org/10.1016/j.jallcom.2016.09.207 A. Ezhilarasi, J. Judith Vijaya, K. Kaviyarasu, M. Maaza, A. Ayeshamariam, L. John Kennedy, Green synthesis of NiO nanoparticles using moringo oleifera extract and their nanoparticles biomedical applications: Cytotoxicity effect of nanoparticles against Ht29 cancer cells, J. Photochem. Photobiol. B: Biology (2016) 164 352–360. https://doi.org/10.1016/j.jphotobiol.2016.10.003 A.E. Nel, L. Madler, D. Velegol, T. Xia, E.M.V. Hoek, P. Somasundaram, F. Klaessig, V. Castranova, M. Thompson, Nat. Mater (2009) 8(7):543-57. https://doi.org/10.1038/nmat2442 , A. El-Trass, H. ElShamy, I. El-Mehasseb, M. El-Kemary. CuO Nanoparticles: Synthesis, Characterization, Optical Properties and Interaction with Amino Acids. Applied Surface Science ( 2012) , 258 (7), 2997–3001. https://doi.org/10.1016/j.apsusc.2011.11.025 M. El-Kemary, N. Nagy, and I. El-Mehasseb. Nickel oxide nanoparticles: Synthesis and spectral studies of interactions with glucose. Materials Science in Semiconductor Processing 16(6) 1747-1752. https://doi.org/10.1016/j.mssp.2013.05.018 M. Pudukudy, Z. Yaakob. Facile Synthesis of Quasi Spherical ZnO Nanoparticles with Excellent Photocatalytic Activity. J Clust Sci ( 2015) , 26 (4), 1187–1201. https://doi.org/10.1007/s10876-014-0806-1 K. Borgohain, J. Singh, M. Rama Rao, T. Shripathi, S. Mahamuni, Quantum size effects in CuO nanoparticles, Phys. Rev. B. (2000) 61 11093–11096. https://doi.org/10.1103/PhysRevB.61.11093. T. Ivanova, A. Harizanova, T. Koutzarova, B. Vertruyen, Study of ZnO sol-gel films: Effect of annealing, Mater. Lett. (2010) 64 1147–1149. https://doi.org/10.1016/j.matlet.2010.02.033 Z. Yang, Z. Ye, Z. Xu, B. zhao, Effect of the morphology on the optical properties of ZnO nanostructures, Phys. E Low-Dimensional Syst. Nanostructures. (2009) 42 116– 119. https://doi.org/10.1016/j.physe.2009.09.010 K. Anandan, V. Rajendran, Morphological and size effects of NiO nanoparticles via solvothermal process and their optical properties, Mater. Sci. Semicond. Process. (2011) 14 43–47. https://doi.org/10.1016/j.mssp.2011.01.001 W.-Q. Wu, B.-X. Lei, H.-S. Rao, Y.-F. Xu, Y.-F. Wang, C.-Y. Su, D.-B. Kuang. Hydrothermal Fabrication of Hierarchically Anatase TiO2 Nanowire Arrays on FTO Glass for Dye-Sensitized Solar Cells. Sci Rep ( 2013) , 3 (1), 1352. https://doi.org/10.1038/srep01352 Juma, A. O.; Arbab, E. A. A.; Muiva, C. M.; Lepodise, L. M.; Mola, G. T. Synthesis and Characterization of CuO-NiO-ZnO Mixed Metal Oxide Nanocomposite. Journal of Alloys and Compounds 2017 , 723 , 866–872. https://doi.org/10.1016/j.jallcom.2017.06.288 M.A. Subhan, T. Ahmed, N. Uddin, Synthesis, structure, PL and photocatalytic activities of La2O2CO3.CeO2.ZnO fabricated by coprecipitation method, Spectrochimic. Acta Part A: Mol. Biomol. Spectrosc (2015) 138, 827-833. https://doi.org/10.1016/j.saa.2014.10.114 M.A. Subhan, T. Ahmed, M.R. Awal, A.M.M. Fahim, Synthesis, structure and excitation wavelength dependent PL properties of novel nanocomposite La2O2CO3.CuO.ZnO, Spectrochimic. Acta Part A: Mol. Biomol. Spectrosc (2014) 132, 550-554 https://doi.org/10.1016/j.saa.2014.05.015 M.A. Subhan, T. Ahmed, Synthesis, characterization and spectroscopy investigations of novel nano multi-metal oxide Co3O4. CeO2.ZnO, Spectrochimic. Acta Part A: Mol. Biomol. Spectrosc (2014) 129, 377-381. https://doi.org/10.1016/j.saa.2014.03.051 M.A. Subhan, T. Ahmed, N. Uddin, A.K. Azad, K. Begum, Synthesis, characterization, PL properties, photocatalytic and antibacterial activities of nano multi-metal oxide NiO.CeO2.ZnO, Spectrochimic. Acta Part A. Mol. Biomol. Spectrosc (2015) 136, 824-831. https://doi.org/10.1016/j.saa.2014.09.100 K. Karthik, S. Dhanuskodi, C. Gobinath . Multifunctional properties of microwave assisted CdO–NiO–ZnO mixed metal oxide nanocomposite: enhanced photocatalytic and antibacterial activities. J Mater Sci: Mater Electron (2018) 29(7) , 5459–5471. https://doi.org/10.1007/s10854-017-8513-y M. Hafeez, R. Arshad, J. Khan, B. Akram, M. N. Ahmad, M. U. Hameed, S. Haq. Populus ciliata mediated synthesis of copper oxide nanoparticles for potential biological applications. Mater. Res. Express. (2019) 6(5), 055-043. https://doi.org/10.1088/2053-1591/ab0601 D. Paul, S. Mangla, S. Neogi. Antibacterial study of CuO-NiO-ZnO trimetallic oxide nanoparticle. Mater. Lett. (2020) , 271, 127740. https://doi.org/10.1016/j.matlet.2020.127740 L. Tao, D. Dastan, W. Wang, P. Poldorn, X. Meng, M. Wu, H. Zhao, H. Zhang, L. Li, B . An. Metal-Decorated InN Monolayer Senses N2 against CO2. ACS Appl. Mater. Interfaces (2023), 15, 12534− 12544. https://doi.org/10.1021/acsami.2c21463 D. S. Ahmed, M.K.A. Mohammed. Studying the bactericidal ability and biocompatibility of gold and gold oxide nanoparticles decorating on multi-wall carbon nanotubes. Chem. Papers (2020), 74, 4033−4046. https://doi.org/10.1007/s11696-020-01223-0 B. Luan, C. Liu, P. Chen, and R. Chen. "Nanotechnology Reviews “Just Accepted” Papers Nanotechnol." (2011). M. KA. Mohammed, Ali K. Al-Mousoi, S. M. Majeed, S. Singh, A. Kumar, R. Pandey, J. Madan, D. S. Ahmed, and D. Dastan. Stable hole-transporting material-free perovskite solar cells with efficiency exceeding 14% via the introduction of a malonic acid additive for a perovskite precursor. Energy & Fuels (2022) 36(21) 13187-13194. https://doi.org/10.1021/acs.energyfuels.2c02878 M. KA. Mohammed, Ali K. Al-Mousoi, S. M. Majeed, S. Singh, A. Kumar, R. Pandey, J. Madan, D. S. Ahmed, and D. Dastan. Ionic Liquid Passivator for Mesoporous Titanium Dioxide Electron Transport Layer to Enhance Efficiency and Stability of Hole Conductor-Free Perovskite Solar Cells. Energy Fuels (2022), 36(19), 12192−12200. https://doi.org/10.1021/acs.energyfuels.2c01980 B. Akram, K. Ahmad, J. Khan, B. A. Khan, and J. Akhtar. Low-temperature solution-phase route to sub-10 nm titanium oxide nanocrystals having super-enhanced photoreactivity. New Journal of Chemistry (2018) 42(13) 10947-10952. https://doi.org/10.1039/C8NJ02317D P. Zhu, D. Dastan, L. Liu, L. Wu, Z. Shi, Q.-Q. Chu, F. Altaf, and M. KA Mohammed. Surface wettability of various phases of titania thin films: Atomic-scale simulation studies. Journal of Molecular Graphics and Modelling (2023) 118 108335. https://doi.org/10.1016/j.jmgm.2022.108335 I. Ashraf, S. Ahmad, F. Nazir, D. Dastan, Z. Shi, H. Garmestani, and M. Iqbal. Hydrothermal synthesis and water splitting application of d-Ti3C2 MXene/V2O5 hybrid nanostructures as an efficient bifunctional catalyst. International Journal of Hydrogen Energy (2022) 47(64) 27383-27396. https://doi.org/10.1016/j.ijhydene.2022.06.095 J. Dlugaszewska, R. Dobrucka. Effectiveness of biosynthesized trimetallic Au/Pt/Ag nanoparticles on planktonic and biofilm Enterococcus faecalis and Enterococcus faecium forms. J. Clust. Sci. (2019), 30, 1091−1101. https://doi.org/10.1007/s10876-019-01570-3 G. L. Tan, D. Tang, D. Dastan, A. Jafari, Z. Shi, Q.-Q. Chu, J. PB Silva, and X.-T. Yin. Structures, morphological control, and antibacterial performance of tungsten oxide thin films. Ceramics International (2021) 47(12) 17153-17160. https://doi.org/10.1016/j.ceramint.2021.03.025 R. Mohammadi-Aloucheh, A. Habibi-Yangjeh, A. Bayrami, S. Latifi-Navid, and A. Asadi. Green synthesis of ZnO and ZnO/CuO nanocomposites in Mentha longifolia leaf extract: characterization and their application as anti-bacterial agents. Journal of Materials Science: Materials in Electronics (2018) 29(16) 13596-13605. https://doi.org/10.1007/s10854-018-9487-0 M. Nasrollahzadeh, M. Sajjadi, S. Iravani, and R. S. Varma. Trimetallic nanoparticles: greener synthesis and their applications. Nanomaterials (2020) 10(9) 1784. https://doi.org/10.3390/nano10091784 V. Manikandan, P. Velmurugan, J. H. Park, N. Lovanh, S. K. Seo, P. Jayanthi, Y. J. Park, M. Cho, and B. T. Oh. Synthesis and antimicrobial activity of palladium nanoparticles from Prunus× yedoensis leaf extract. Materials Letters (2016) 185 335-338. https://doi.org/10.1016/j.matlet.2016.08.120 S. Srihasam, K. Thyagarajan, M. Korivi, V. R. Lebaka, and S. P. R. Mallem. Phytogenic generation of NiO nanoparticles using Stevia leaf extract and evaluation of their in-vitro antioxidant and antimicrobial properties. Biomolecules (2020) 10(1) 89 https://doi.org/10.3390/biom10010089 F. Amin, B. Khattak, A. Alotaibi, M. Qasim, I. Ahmad, R. Ullah, M. Bourhia, A. Gul, S. Zahoor, R. Ahmad. Green synthesis of copper oxide nanoparticles using Aerva javanica leaf extract and their characterization and investigation of in vitro antimicrobial potential and cytotoxic activities. Evidence-Based Complement. Altern. Med. (2021) , 2021, 1−12. https://doi.org/10.1155/2021/5589703 D. Suresh, P. C. Nethravathi, H. Rajanaika, H. Nagabhushana, and S. C. Sharma. "Green synthesis of multifunctional zinc oxide (ZnO) nanoparticles using Cassia fistula plant extract and their photodegradative, antioxidant and antibacterial activities." Materials Science in Semiconductor Processing (2015) 31 446-454. https://doi.org/10.1016/j.mssp.2014.12.023 F. Sowmiya, , S. R. Brintha. Synthesis and Characterization of Zno-Cdo Nanocomposite and Its Antibacterial Activity. J. Pharm. Negat. Results (2022), 13, 3559−3569. W. Chen, L. Yue, Q. Jiang, X. Liu, and W. Xia.Synthesis of varisized chitosan-selenium nanocomposites through heating treatment and evaluation of their antioxidant properties. Int. J. Biol. Macromol. (2018), 114, 751−758. https://doi.org/10.1016/j.ijbiomac.2018.03.108 B.V. Vibitha , B. Anitha, P. G. A. Krishna, and J. N. Tharayil. Plant extracts assisted synthesis, characterization and antioxidant properties of ZnO: CuO nanocomposites. In AIP Conference Proceedings , vol. 2244(1), p. 070037. AIP Publishing LLC, 2020. https://doi.org/10.1063/5.0009174 S. Hajji, R. B. S.-B. Salem, M. Hamdi, K. Jellouli, W. Ayadi, M. Nasri, and S. Boufi. Nanocomposite films based on chitosan–poly (vinyl alcohol) and silver nanoparticles with high antibacterial and antioxidant activities. Process Safety and Environmental Protection (2017) 111 112-121. https://doi.org/10.1016/j.psep.2017.06.018 P. Siriwong, T. Thongtem, A. Phuruangrat, and S. Thongtem. Hydrothermal synthesis, characterization, and optical properties of wolframite ZnWO 4 nanorods. CrystEngComm (2011): 13(5) 1564-1569. https://doi.org/10.1039/C0CE00402B [55 K. Karthik, S. Dhanuskodi, C. Gobinath . Multifunctional properties of microwave assisted CdO–NiO–ZnO mixed metal oxide nanocomposite: enhanced photocatalytic and antibacterial activities. J Mater Sci: Mater Electron (2018) 29(7) , 5459–5471. https://doi.org/10.1007/s10854-017-8513-y M.A. Subhan, T. Ahmed, N. Uddin, A.K. Azad, K. Begum, Synthesis, characterization, PL properties, photocatalytic and antibacterial activities of nano multi-metal oxide NiO.CeO2.ZnO, Spectrochimic. Acta Part A. Mol. Biomol. Spectrosc (2015) 136, 824-831. https://doi.org/10.1016/j.saa.2014.09.100 M.A. Subhan, N. Uddin, P. Sarker, H. Nakata, R. Makioka, Synthesis, characterization, low temperature solid state PL and photocatalytic activities of Ag2O, CeO2.ZnO nanocomposite, Spectrochimic. Acta Part A. Mol. Biomol. Spectrosc (2015) 151, 56-63 https://doi.org/10.1016/j.saa.2015.06.094 T.K. Jana, S.K. Maji, A. Pal, R.P. Maiti, T.K. Dolai, K. Chatterjee, Photocatalytic and antibacterial activity of cadmium sulphide/zinc oxide nanocomposite with varied morphology. J. Colloid Interf. Sci (2016) 480, 9-16 https://doi.org/10.1016/j.jcis.2016.06.073 K. Ravichandran, N. Chidhambaram, S. Gobalakrishnan, Copper and graphene activated ZnO nanoparticles for enhanced photocatalytic and antibacterial activities. J. Phys. Chem. Solids (2016) 93, 82-90 https://doi.org/10.1016/j.jpcs.2016.02.013 N. Padmavathy, R. Vijayaraghavan . Interaction of ZnO nanoparticles with microbes – a physio and biochemical assay. J Biomed Nanotechnol. (2011) 7(6) 813–822 https://doi.org/10.1166/jbn.2011.1343 J. Yu, W. Zhang, Y. Li. Synthesis, characterization, antimicrobial activity and mechanism of a novel hydroxyapatite whisker/nano zinc oxide biomateriali. Biomed Mater. (2014) 10(1):015001. https://doi.org/10.1088/1748-6041/10/1/015001. W. Qiu, X. Zheng, Y. Wei. D-alanine metabolism is essential for growth and biofilm formation of Streptococcus mutans. Mol Oral Microbiol. (2016) 31(5), 435–444. https://doi.org/10.1111/omi.12146 J. Iqbal, B. A. Abbasi, A. Munir, S. Uddin, S. Kanwal, and T. Mahmood. Facile green synthesis approach for the production of chromium oxide nanoparticles and their different in vitro biological activities. Microscopy research and technique (2020) 83(6) 706-719. https://doi.org/10.1002/jemt.23460 J. Stetefeld, S. A. McKenna, and T. R. Patel, Dynamic light scattering: a practical guide and applications in biomedical sciences. Biophysical reviews. (2016) 8(4) 409-27. https://doi.org/10.1007/s12551-016-0218-6 Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7532947","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":516021947,"identity":"14398b85-a8f4-40c6-b6d0-05b1ffa5d1e6","order_by":0,"name":"Vikas Choudhary","email":"","orcid":"","institution":"Central University of Himachal Pradesh","correspondingAuthor":false,"prefix":"","firstName":"Vikas","middleName":"","lastName":"Choudhary","suffix":""},{"id":516021948,"identity":"2d0bd1e0-f101-4e66-b4b1-4b30406a24a2","order_by":1,"name":"Kusham Lata","email":"","orcid":"","institution":"Central University of Himachal Pradesh","correspondingAuthor":false,"prefix":"","firstName":"Kusham","middleName":"","lastName":"Lata","suffix":""},{"id":516021949,"identity":"89f01632-0285-4d3e-bdc0-ba247d0c01c0","order_by":2,"name":"Manish Kumar","email":"","orcid":"","institution":"Central University of Himachal Pradesh","correspondingAuthor":false,"prefix":"","firstName":"Manish","middleName":"","lastName":"Kumar","suffix":""},{"id":516021950,"identity":"e8aff59d-a838-4f93-9569-29a1eacbd604","order_by":3,"name":"Ajay Sharma","email":"","orcid":"","institution":"Career Point University","correspondingAuthor":false,"prefix":"","firstName":"Ajay","middleName":"","lastName":"Sharma","suffix":""},{"id":516021951,"identity":"e4ea7d38-86a1-47a3-96da-3aa8eb4d1795","order_by":4,"name":"Raman Kumar","email":"","orcid":"","institution":"MMEC, Maharishi Markandeshwar (Deemed to be University)","correspondingAuthor":false,"prefix":"","firstName":"Raman","middleName":"","lastName":"Kumar","suffix":""},{"id":516021952,"identity":"a3cb031c-bb44-4678-bb28-098352e88157","order_by":5,"name":"Vivek Sheel Jaswal","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1klEQVRIiWNgGAWjYDACCSBmbGBg4GMHEgwGFiRoYeM5ANIiQYoWiQQYlwDgn918dOPPHTZybJLPr274USDBwN/enYDfkjvH0m7znkkzZpPOKbvZA3SYxJmzG/BqMZDIMbvN2HY4sU06J+0GD1CLgUQuIS35327+bPtf3yZ5Ju3mH+K05LDd4G07kMAmwX7sNlG2SNxIM7vN25Zs2MaTw3ZbxkCCh6Bf+GckPwM6zE6en/34s5tv/tjI8bf34teCBHgMwCSxykGA/QEpqkfBKBgFo2AEAQCFckYCXvDRlgAAAABJRU5ErkJggg==","orcid":"","institution":"Central University of Himachal Pradesh","correspondingAuthor":true,"prefix":"","firstName":"Vivek","middleName":"Sheel","lastName":"Jaswal","suffix":""}],"badges":[],"createdAt":"2025-09-04 06:53:27","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7532947/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7532947/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":91562132,"identity":"424e0151-ae11-45b5-ae2f-300e1ecc3a0e","added_by":"auto","created_at":"2025-09-17 18:52:16","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":241611,"visible":true,"origin":"","legend":"\u003cp\u003eUV-Visible Spectra of (a) CuO-NiO-ZnO nanocomposite annealed at 200\u003csup\u003eº\u003c/sup\u003eC \u0026amp; Tauc-plot showing band-gap graph at 200\u003csup\u003eº\u003c/sup\u003eC (b) CuO-NiO-ZnO nanocomposite annealed at 500\u003csup\u003eº\u003c/sup\u003eC \u0026amp; Tauc-plot showing band-gap graph at 500\u003csup\u003eº \u003c/sup\u003eC (c) Particle size distribution CuO-NiO-ZnO Nanocomposite at 200\u003csup\u003eº\u003c/sup\u003eC (d) Particle size distribution CuO-NiO-ZnO Nanocomposite at 500\u003csup\u003eº\u003c/sup\u003eC\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-7532947/v1/98daa8d6807d6d39e817b5b9.png"},{"id":91562134,"identity":"99f04935-d76b-4698-86ee-88e710e6eb74","added_by":"auto","created_at":"2025-09-17 18:52:16","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":647636,"visible":true,"origin":"","legend":"\u003cp\u003e(a) X-ray diffraction pattern of CuO-ZnO-NiO NCs annealed at 200\u003csup\u003eO\u003c/sup\u003eC \u0026amp; 500\u003csup\u003eO\u003c/sup\u003eC (b) FE SEM images of CuO-ZnO-NiO NCs at 200\u003csup\u003eO\u003c/sup\u003eC. (c) Histogram showing size distribution of CuO-ZnO-NiO NCs at 200\u003csup\u003eO\u003c/sup\u003eC. (d) FE SEM images of CuO-ZnO-NiO NCs at 500\u003csup\u003eO\u003c/sup\u003eC (e) Histogram showing size distribution of CuO-ZnO-NiO NCs at 500\u003csup\u003eO\u003c/sup\u003eC. (f) FTIR Spectra of CuO-ZnO-NiO NCs annealed at 200\u003csup\u003eO\u003c/sup\u003eC \u0026amp; 500\u003csup\u003eO\u003c/sup\u003eC\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-7532947/v1/98ba983b8f7f5b2f0b7de50b.png"},{"id":91562133,"identity":"8016802f-0f94-47fd-a6b1-0fc529e41b30","added_by":"auto","created_at":"2025-09-17 18:52:16","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":688556,"visible":true,"origin":"","legend":"\u003cp\u003eThe antibacterial action of CuO-NiO-ZnO nanomaterials against \u003cem\u003eBacillus subtilis\u003c/em\u003e, \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e, \u0026amp; \u003cem\u003eEscherichia coli\u003c/em\u003e. In Figure 3 i ( a) showed inhibition zones against \u003cem\u003eB. subtilis\u003c/em\u003e (gram positive ) bacteria at different concentration (b) showed inhibition zones against \u003cem\u003eP. aeruginosa \u003c/em\u003e\u0026nbsp;(GPB) at different concentration\u0026nbsp; ( c)\u0026nbsp; showed inhibition zones against \u003cem\u003eE.coli\u0026nbsp; \u003c/em\u003e\u0026nbsp;(GNB) at different concentration indicates CuO-NiO-ZnO at 200\u003csup\u003eO\u003c/sup\u003eC showed larger inhibition zones against \u003cem\u003eE. coli\u003c/em\u003e, indicating greater effectiveness against Gram-negative bacteria. Figure 3(ii) quantitatively confirmed increased inhibition with rising CuO-NiO-ZnO at 200\u003csup\u003eO\u003c/sup\u003eC concentrations against (\u003cem\u003eB. subtilis, P. aeruginosa,\u003c/em\u003e \u0026amp; \u003cem\u003eE.coli).\u003c/em\u003e Figure 3iii (a, b, c) (a) showed inhibition zones against \u003cem\u003eB. subtilis\u003c/em\u003e (gram-positive) bacteria at different concentrations, (b) showed inhibition zones against \u003cem\u003eP. aeruginosa \u003c/em\u003e\u0026nbsp;(GPB) at different concentrations, and (c) showed inhibition zones against \u003cem\u003eE.coli\u003c/em\u003e (GNB) at different concentrations. This indicates enhanced inhibition zones with CuO-NiO-ZnO at 500\u003csup\u003eO\u003c/sup\u003eC, particularly for \u003cem\u003eB. subtilis\u003c/em\u003e. Figure 3(iv) quantitatively revealed that CuO-NiO-ZnO at 500\u003csup\u003eO\u003c/sup\u003eC had a stronger effect on Gram-positive bacteria, with inhibition increasing dose-dependently. Overall, CuO-NiO-ZnO at 200\u003csup\u003eO\u003c/sup\u003eC was more effective against Gram-negative strains, while CuO-NiO-ZnO at 500\u003csup\u003eO\u003c/sup\u003eC showed improved activity toward Gram-positive bacteria.\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-7532947/v1/16a698f5d4134bb9e0ed189d.png"},{"id":91562138,"identity":"bf363146-b511-443d-8e7c-a962f26b6ec9","added_by":"auto","created_at":"2025-09-17 18:52:16","extension":"jpeg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":149342,"visible":true,"origin":"","legend":"\u003cp\u003eMechanism of antibacterial activity of ZnO-NiO-CuO NC\u003c/p\u003e","description":"","filename":"image5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7532947/v1/eb459d4d471885dfa0c6280f.jpeg"},{"id":91565577,"identity":"a0b97a80-d040-42e1-a7ea-34aad172759a","added_by":"auto","created_at":"2025-09-17 19:24:17","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3026512,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7532947/v1/16288f2b-ab11-4a96-9538-0484ad9d7fa8.pdf"},{"id":91562820,"identity":"d8c2ff4b-cb57-4a61-b840-b24ade293d5d","added_by":"auto","created_at":"2025-09-17 19:00:16","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":657209,"visible":true,"origin":"","legend":"","description":"","filename":"GraphicalAbstract.docx","url":"https://assets-eu.researchsquare.com/files/rs-7532947/v1/e34ea614fcbd63f5fe9fe3ae.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Antibacterial Application of Heterogeneous CuO-NiO-ZnO Metal Oxides Nanocomposites","fulltext":[{"header":"Highlights","content":"\u003cul\u003e\n \u003cli\u003eSynthesis of CuO-NiO-ZnO nanocomposites by the co-precipitation method.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eCharacterization techniques used for the assurance of CuO-NiO-ZnO nanocomposites formation were UV-visible, FTIR, XRD, DLS, and SEM analysis.\u003c/li\u003e\n \u003cli\u003eEnhanced antibacterial efficacy of\u0026nbsp;CuO-NiO-ZnO against \u003cem\u003eBacillus\u003c/em\u003e,\u0026nbsp;\u003cem\u003ePseudomonas\u003c/em\u003e \u0026amp; \u003cem\u003eE. coli\u0026nbsp;\u003c/em\u003ebacterial strains.\u003c/li\u003e\n\u003c/ul\u003e"},{"header":"1. Introduction","content":"\u003cp\u003eThe scientific fields of engineering, chemistry, physics, and materials science have experienced an increased interest in the development of applications and research on mixed metal oxides. Combining multiple metals together in the oxide framework can develop materials having unique chemical \u0026amp; physical characteristics that result in comparatively better performance in wide range of applications. The metals may exhibit the characteristics of isolated entities that contribute their qualities to the system, or they may have altered properties by interactions between metals or between metals and oxygen [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. A nanostructured system's performance can be significantly enhanced by the interaction of its many constituent materials. At the microscopic level, the close contact surfaces of composite materials with corresponding band potentials improve the efficiency of charge separation, lengthen the life of charge carriers, \u0026amp; improve transfer of charge [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Most of the mixed oxides (MO) are single-phase materials that can reveal functional properties resulting from combining individual oxide functionalities. Multiphase nanocomposites, created by the coexistence of two MO within the same surrounding substance, have been made for applications such as photocatalysis \u0026amp; antibacterial application [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Various metal-oxide nanocomposites, including MnOx- CeO₂ [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e], ZnO- CuO [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], NiO- ZnO [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], and MgO- ZnO [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e], were synthesized and studied using a variety of methodologies.\u003c/p\u003e\u003cp\u003eAmongst these, the widely utilized composite MnOx supported on Ti\u003csub\u003e6\u003c/sub\u003eAl\u003csub\u003e4\u003c/sub\u003eV has been synthesized, which has influenced its properties. According to these studies, composite materials trap photogenerated carriers more effectively than monomeric oxides. A few studies have shown enhanced device performance and characteristics for MO NCs with three or more phases, such as ZnO-WO₃-ZnWO₃ [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e], Mo-W-V MOs [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], La-V-Mo oxide solid solution [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], and the pseudo-quaternary system CaO-SiO₂-CoO [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Some conclusions have been reported by another investigation on \"ZnO-CeO\u003csub\u003e2\u003c/sub\u003e-TiO₂\" composites made by combustion, which revealed improved photocatalytic color removal and outstanding UV absorption. [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e] Infectious disorders caused by microorganisms like fungi, viruses, parasites, or bacteria have recently affected public health in many countries and are one of the leading causes of mortality worldwide. Because of their electron-donating qualities, metallic oxide-composited components are known to encourage the production of the activated oxygen class, according to researchers. Researchers have recently discovered the antibacterial properties of different metal oxide NCs [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. There have been reports of nanocomposites targeted at both Gram-positive and Gram-negative microorganisms. The targeting of the bacteria was identified through an electrostatic interaction between the positive charge residues on the composite surface and the negative charge microbial membrane.\u003c/p\u003e\u003cp\u003eCopper oxides NPs have attracted significant attention among other metal oxide nanoparticles. CuO is a low-bandgap (1.2 eV) p-type semiconductor that finds utility as a high-Tc superconductor in sensing gas, photocatalysis, photovoltaics, magnetic storage, \u0026amp; technology. By preventing the development of fungi, viruses, bacteria, \u0026amp; algae, CuO NPs in particular exhibit strong antibacterial qualities. When added to coatings, copper oxide nanoparticles (CuO NPs) can function as antifungal and antibacterial agents. [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn recent years, nickel oxide (NiO) which is a p-type semiconductor with a broadband gap of 3.8eV (3.6\u0026ndash;4.0 eV), has garnered a lot of interest because of its uses in gas sensoring application, pigments, spin valve devices, alkaline battery cathodes, etc. [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e] NiO is regarded as an appropriate material for solar cells, antiferromagnetic layers, lithium-ion battery anodes, and other applications because of its high theoretical capacity, quick reaction kinetics, \u0026amp; strong cyclic reversibility [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. NiO is extremely sensitive to a wide range of gases, including ethanol, CO, NO\u003csub\u003e2\u003c/sub\u003e, ammonia, and acetone [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. High chemical stability, exceptional capacity of electron transfer, \u0026amp; anti-inflammatory qualities are all possessed by NiO nanoparticles. Their special qualities, such as release of metal ion, surface area, \u0026amp; adsorbing capacity, also provide them with cytotoxic effects [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe studies on zinc oxide nanostructures are gaining attentions because of their distinctive characteristics and numerous uses. Using ZnO nanoparticles has the benefit of significantly reducing the activity of harmful bacteria at low concentrations. Since ZnO nanoparticles have antifungal \u0026amp; antibacterial properties at lower concentrations, their thin coatings can be utilized for developing products that are resistant to bacteria. ZnO is a member of the metal oxide class, which is distinguished by its ability to photocatalyze and photooxidize biological and chemical species. It makes use of a multipurpose nano platform that releases reactive oxygen species (ROS) to attack cancerous cells from the outside [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. ZnO nanoparticles' antibacterial effects result from an electrical interaction between the greater cell damage as a result of the nanoparticles' increased interaction with the cell surface. Previous studies showed that the wavelength-dependent fluorescence of excitation by CuO.La₂O₂Co₃.ZnO [28], Co₃O₄.CeO₂.ZnO [29], and La₂O₂Co₃.CeO₂.ZnO [27] nanocomposites has been studied by scientists. When exposed to methyl violet 6b (MV), the La₂O₂Co₃.CeO₂.ZnO composite exhibits the 93.75% efficiency of photocatalytic degradation. For the same dye, the NiO.CeO₂.ZnO [30] composite shows 96.07% efficiency. It also demonstrates powerful antimicrobial activity against harmful bacteria such as P. mirabilis, with a zone of inhibition (ZOI) of 15 mm. Ag₂O, CeO₂, and ZnO exhibit antibacterial action against K. pneumoniae with a ZOI of 15 mm \u0026amp; a photocatalytic destruction rate of 84.04% against MV. Scientists have reported the synthesis \u0026amp; characterization of CdO\u0026ndash;ZnO\u0026ndash;NiO mixed MO nanocomposite for the photocatalytic \u0026amp; antibacterial activities of the microwave-assisted nanocomposite [31].\u003c/p\u003e\u003cp\u003eThe combination of these three distinct metallic-oxide phases forms a heterogeneously mixed metal oxide \u0026ldquo;CuO-NiO-ZnO nanocomposite\u0026rdquo; that provides a range of options for controlling the nanocomposite's characteristics \u0026amp; defining novel technological applications in the antibacterial activity for targeting a wide variety of hazardous microbes.\u003c/p\u003e"},{"header":"2. Experimental Details","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1 Materials\u003c/h2\u003e\u003cp\u003e\u003cstrong\u003eMaterials and methods\u003c/strong\u003e\u003cp\u003eAll chemicals used were of analytical grade and used without further purification. Nickel nitrate hexahydrate (Ni (NO\u003csub\u003e3\u003c/sub\u003e)\u003csub\u003e2\u003c/sub\u003e.6H\u003csub\u003e2\u003c/sub\u003eO), Copper Sulphate Pentahydrate (CuSO4.5H2O), zinc chloride (ZnCl2), Sodium Hydroxide was purchased from LOBA CHEMIE PVT, Ethanol. Bacterial strains were initially procured from the Institute of Microbial Technology in Chandigarh, India.\u003c/p\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2 Synthesis of the Nanocomposites\u003c/h2\u003e\u003cp\u003eCuO-NiO-ZnO nanocomposite was synthesized via the co-precipitation method.\u003c/p\u003e\u003cp\u003eA 0.01 M predecessor solution of three salts was prepared by weighing 300 mg of Cu, 400 mg of Ni, and 500 mg of Zn salts, and then mixing it with the 50 ml solution containing 40 ml of water with 10 ml of ethanol. The solution was swirled magnetically at temperature of 50\u0026deg;C for time period of 30 minutes. To make a 50 ml mixed solvent of NaOH, two grams of NaOH pellets were dissolved in five minutes of distilled water. The solution of NaOH is then added to the combined precursor solution with continuously stirring of solution. A precipitate is seen to form immediately after the color of mixture changes from bluish-green to a shade of blackish-green. After five minutes, the temperature is raised to 100℃ \u0026amp; maintained there for an additional half-hour while being continuously agitated. The mixture was then let to cool at room temperature. Filtration method was utilized to collect the precipitate, which was then washed three times with distilled water \u0026amp; dried on a hot plate at 100℃ for two hours. A fine black powder is then produced by annealing the powder for two hours at 200 and 500 ℃ [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3 Characterizations\u003c/h2\u003e\u003cp\u003eThe properties of the synthesized materials (CuO-NiO-ZnO nanocomposite) were thoroughly investigated using various characterization techniques. Fourier Transform Infrared (FTIR) spectroscopy (Perkin Elmer, subtech spectrum) was employed to classify the functional groups within CuO-NiO-ZnO nanocomposite in the 4000\u0026thinsp;\u0026minus;\u0026thinsp;400 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e spectral range. Powder X-ray Diffraction (XRD) analysis using a PAN analytical EMPEREAN diffractometer with Cu K alpha radiation (λ\u0026thinsp;=\u0026thinsp;1.5405980 \u0026Aring;) provided insights into the crystallinity, phase composition, and crystallite size of the materials. Field Emission Scanning Electron Microscopy (FESEM) \u0026amp; Energy Dispersive Spectroscopy (EDS) were utilized to investigate the morphology, size, and surface topography, while EDS analysis provided composition data. UV-visible spectroscopy (Specord 200 PLUS) was employed to determine optical properties, including band gap estimation. Dynamic light scattering (DLS) is a quasi-elastic light dispersion technique which is used to find out the size distribution of selective NPs and is also used for evaluation of the size \u0026amp; dispersion of the nanocomposite.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e\u003cb\u003e2.4 Determination of antibacterial activity\u003c/b\u003e\u003c/h2\u003e\u003cp\u003eThe antibacterial property of CuO-ZnO-NiO nanocomposite was evaluated using the agar well diffusion method against two GNB, \u003cem\u003eEscherichia coli\u003c/em\u003e (MTCC 1652), \u0026amp; \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e (MTCC 741), as well as one GPB, \u003cem\u003eBacillus subtilis\u003c/em\u003e (MTCC 121). The Institute of Microbial Technology in Chandigarh, India, was the original source of the bacterial strains. Overnight cultures of the bacteria were diluted to approximately 10\u003csup\u003e8\u003c/sup\u003e CFU/mL using sterile distilled water, \u0026amp; 1 mL of the diluted inoculum was spread onto nutrient agar plates (25 mL per plate). Six to eight wells (7 to 8 mm in diameter) were punched into the agar using a sterilized cork borer. The wells were then filled with 1 mL of freshly prepared CuO-NiO-ZnO nanocomposite solutions at concentrations ranging between 0.05 \u0026micro;L to 5 \u0026micro;L for the first screening, and 1 mg/mL for the subsequent evaluation. Dimethyl sulfoxide (DMSO) was used as a solvent for the test compounds and also functioned as a negative control. Plates are incubated at a temperature of 37\u0026deg;C for a time period of 24 \u0026amp; 48 hours, and the zone of inhibition is measured in millimeters to assess the antimicrobial activity. Bacitracin \u0026amp; Chloramphenicol (10 mg/mL) were used as positive controls. In order to assess the antibacterial potential of the nanocomposites, the inhibitory zones surrounding the wells were inspected and documented after the incubation period.\u003c/p\u003e\u003cp\u003eThe modified agar well diffusion method is utilized to test the minimum inhibitory concentration (MIC) of each compound, giving an inhibitory zone at a concentration of 1 mg/mL. Several wells in the agar plates received various applications of a single chemical at different concentrations (1,000\u0026ndash;1 \u0026micro;g/mL). Each dilution was added to wells at a volume of about 1 mL. During 48 hours, all test plates were incubated at 37\u0026deg;C. A definite zone of inhibition was visible in the lowest concentration of each chemical, which was referred to as the MIC.\u003c/p\u003e\u003c/div\u003e"},{"header":"3. Results and discussion","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003e3.1 UV\u0026ndash;visible analysis:\u003c/h2\u003e\u003cp\u003eUV-visible absorption spectra \u0026amp; Tauc plots of the synthesized mixed MO nanocomposites at 200 and 500 ℃ as represented in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e(a \u0026amp; b). The UV region, which ranges from 200 to 400 nm, is where the distinctive peaks in the Ultraviolet-Visible analysis can be observed. The graph shows a prominent SPR absorbance peak at a specific wavelength (λmax) of 288nm \u0026amp; 296nm, confirming the synthesis of CuO NPs upon reduction [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. On the other hand, it shows an absorbance peak at a specific wavelength (λmax) of 338nm, indicating the presence of NiO [19]. In the case of ZnO absorbance peak at a specific wavelength (λmax) of 360nm \u0026amp; 372nm [20]. The bandgap of the CuO-ZnO-NiO nanocomposites, which are annealed at a temperature of 200 ℃, was determined to be 2.70 eV with a slight change to 2.67 eV following annealing at 500\u0026deg;C.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003e3.2 Dynamic Light Scattering (DLS):\u003c/h2\u003e\u003cp\u003eDynamic light scattering (DLS) is a quasi-elastic light dispersion technique that is used to evaluate the dispersion \u0026amp; size of NPs as well as to determine the size distribution of selected NPs. [63]. The particle size distribution of the mixed metal oxides of CuO/NiO/ZnO NCs solution, which is annealed at 200\u0026deg;C, is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e(c). Depending upon the results of DLS, 100% of the particles have a diameter of 450.9 nm. In addition, the DLS provides results shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e(d) that 79.0% of the particles have a diameter of 717.1 nm, 182.8 nm by 19.3% of the particles, and 5118 nm by 1.7% of the particles in mixed metal oxides of CuO/NiO/ZnO NCs solution, which is annealed at 500\u0026deg;C. The Poly-dispersity Index (PdI) for mixed metal oxides of CuO/NiO/ZnO NCs solution, which is annealed at 200\u0026deg;C \u0026amp; 500\u0026deg;C, are 0.280 \u0026amp; 0.352, respectively. It represents that both the mixed metal oxides of CuO/NiO/ZnO NCs solution, which are annealed at 200\u0026deg;C \u0026amp; 500\u0026deg;C, respectively, have a particle distribution of 'mono-scattered' order that explains the distribution of particle size, which consists of a one-size mode without an aggregate mode. The PDI having values between 0.1 to 0.7 (both values included) are mostly monodispersive in nature, While PDI value greater than 0.7 causes the broadening of size distribution of macromolecular system in solution \u0026amp; non-monomodal distribution approaches for data analyses should be considered [64].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\u003ch2\u003e3.3 XRD Analysis:\u003c/h2\u003e\u003cp\u003eX-ray diffraction (XRD) spectrum was obtained to investigate the crystallographic structure of the synthesized CuO-NiO-ZnO nanocomposite annealed at 200\u0026deg;C \u0026amp; 500\u0026deg;C in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e (a). Annealing MO composite [CuO-NiO-ZnO] nanoparticles at temperatures of 200 \u0026amp; 500 ℃ changes their XRD properties. Only the peaks corresponding to Zn-O \u0026amp; Cu-O were identifiable after heating to 200\u0026deg;C. Several of the diffraction peaks of the different oxides overlap due to their very similar angles of diffraction. On the other hand, there was no peak of diffraction that was linked to the NiO phase. Because some of the byproducts of the precursors may still be present, Ni (OH) \u003csub\u003e2\u003c/sub\u003e may persist at a temperature of 200\u0026deg;C. Research on NiO-ZnO MO nanoparticles, which were produced in a way similar to that of CuO-ZnO-NiO samples, revealed the existence of ZnO \u0026amp; β-Ni (OH)₂ mixed phases upon annealing at a temperature of 200\u0026deg;C. Previous studies using TG-DSC, XRD, and HRTEM examinations have shown that, depending on the manufacturing procedure, Cu(OH)₂ \u0026amp; Zn(OH)₂ transform into their corresponding oxides at temperatures below 150\u0026deg;C [25] and slightly over 300\u0026deg;C [26], respectively. After being annealed at a temperature of 500\u0026deg;C, the peaks of diffraction for NiO, ZnO, \u0026amp; CuO have been clearly visible, indicating that the remaining hydroxides have fully broken down and pure metal oxides have formed. The end product is a nanocomposite that has a combination of several binary oxide phases.\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\u003eXRD peaks and the corresponding planes obtained from CuO-NiO-ZnO nanocomposites annealed at 200\u0026deg;C \u0026amp; 500\u0026deg;C.\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=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSamples\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2θ (degree)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003ed-spacing (nm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003ePlane (s)\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\u003eCuO-NiO-ZnO\u003c/p\u003e\u003cp\u003e\u003cb\u003e(200\u0026deg;C)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e31.84\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2.808\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e32.81\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2.727\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e110\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e34.47\u003c/p\u003e\u003cp\u003e35.65\u003c/p\u003e\u003cp\u003e36.23\u003c/p\u003e\u003cp\u003e38.98\u003c/p\u003e\u003cp\u003e47.73\u003c/p\u003e\u003cp\u003e54.4\u003c/p\u003e\u003cp\u003e56.68\u003c/p\u003e\u003cp\u003e59.02\u003c/p\u003e\u003cp\u003e62.09\u003c/p\u003e\u003cp\u003e62.85\u003c/p\u003e\u003cp\u003e68.16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2.599\u003c/p\u003e\u003cp\u003e2.516\u003c/p\u003e\u003cp\u003e2.477\u003c/p\u003e\u003cp\u003e2.308\u003c/p\u003e\u003cp\u003e1.903\u003c/p\u003e\u003cp\u003e1.685\u003c/p\u003e\u003cp\u003e1.622\u003c/p\u003e\u003cp\u003e1.563\u003c/p\u003e\u003cp\u003e1.493\u003c/p\u003e\u003cp\u003e1.477\u003c/p\u003e\u003cp\u003e1.374\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e002\u003c/p\u003e\u003cp\u003e111\u003c/p\u003e\u003cp\u003e101\u003c/p\u003e\u003cp\u003e200\u003c/p\u003e\u003cp\u003e102\u003c/p\u003e\u003cp\u003e020\u003c/p\u003e\u003cp\u003e110\u003c/p\u003e\u003cp\u003e202\u003c/p\u003e\u003cp\u003e-113\u003c/p\u003e\u003cp\u003e103\u003c/p\u003e\u003cp\u003e112\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCuO-NiO-ZnO\u003c/p\u003e\u003cp\u003e\u003cb\u003e(500\u0026deg;C)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e31.83\u003c/p\u003e\u003cp\u003e34.6\u003c/p\u003e\u003cp\u003e35.74\u003c/p\u003e\u003cp\u003e36.40\u003c/p\u003e\u003cp\u003e37.14\u003c/p\u003e\u003cp\u003e38.92\u003c/p\u003e\u003cp\u003e43.24\u003c/p\u003e\u003cp\u003e47.74\u003c/p\u003e\u003cp\u003e48.88\u003c/p\u003e\u003cp\u003e56.71\u003c/p\u003e\u003cp\u003e62.81\u003c/p\u003e\u003cp\u003e66.35\u003c/p\u003e\u003cp\u003e68.17\u003c/p\u003e\u003cp\u003e69.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2.809\u003c/p\u003e\u003cp\u003e2.590\u003c/p\u003e\u003cp\u003e2.510\u003c/p\u003e\u003cp\u003e2.466\u003c/p\u003e\u003cp\u003e2.418\u003c/p\u003e\u003cp\u003e2.312\u003c/p\u003e\u003cp\u003e2.090\u003c/p\u003e\u003cp\u003e1.903\u003c/p\u003e\u003cp\u003e1.861\u003c/p\u003e\u003cp\u003e1.621\u003c/p\u003e\u003cp\u003e1.470\u003c/p\u003e\u003cp\u003e1.407\u003c/p\u003e\u003cp\u003e1.358\u003c/p\u003e\u003cp\u003e1.358\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e100\u003c/p\u003e\u003cp\u003e002\u003c/p\u003e\u003cp\u003e111\u003c/p\u003e\u003cp\u003e101\u003c/p\u003e\u003cp\u003e111\u003c/p\u003e\u003cp\u003e200\u003c/p\u003e\u003cp\u003e200\u003c/p\u003e\u003cp\u003e102\u003c/p\u003e\u003cp\u003e-202\u003c/p\u003e\u003cp\u003e110\u003c/p\u003e\u003cp\u003e220\u003c/p\u003e\u003cp\u003e022\u003c/p\u003e\u003cp\u003e112\u003c/p\u003e\u003cp\u003e201\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003e3.4 SEM analysis:\u003c/h2\u003e\u003cp\u003eScanning electron micrographs of the CuO-NiO-ZnO mixed oxide nanocomposites at different magnifications for the samples prepared are presented in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e(b) \u0026amp; (d), annealing at 200\u0026deg;C and 500\u0026deg;C, respectively. The observed shape is spherically irregular \u0026amp; the size distribution observed to be heterogeneous, having some cavities. The morphology of the nanocomposites is observed to vary depending upon the concentration of precursor solution, temperature \u0026amp; many other factors. The average size of particles of nanocomposites at different temperatures (200\u0026deg;C and 500\u0026deg;C) was observed as 49.64nm and 28.96nm, respectively [Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e(c) \u0026amp; (e)]. The sizes of particles calculated in SEM \u0026amp; XRD have some slight variation. The reason for this diminutive difference is that calculations by SEM were based upon the difference between the visible boundaries of grains; on the other hand, calculations of XRD have measured the extended crystalline region, which coherently diffracts X-rays. The image of SEM has shown the aggregated particle, which is due to the excess heat generated during the process of calcination. The size of particles has some minimum changes that occur due to the aggregation of particle.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003e3.5 Fourier Transform Infrared Spectroscopy (FTIR):\u003c/h2\u003e\u003cp\u003eFTIR spectroscopy gives important information on the functional groups and chemical interactions inside the synthesized CuO-NiO-ZnO nanocomposites in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e(f). In the FTIR spectra of synthesized CuO-NiO-ZnO nanocomposites, the distinct characteristic peaks were observed. The absorption bands below 600 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e are mostly linked to the CuO-NiO-ZnO group. Metal-oxygen bond vibrations often exhibit FTIR absorption peaks below 1000 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. CuO has exhibited six infrared vibration bands at 147, 161, 321, 478, 530, \u0026amp; 590 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e [21]. The Zn-O bond vibrational bands may be found at approximately 395, 425, 470, \u0026amp; 515 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, depending upon the structure \u0026amp; shape of the nanoparticles [22, 23]. The IR absorption peaks in NiO are caused by NiO vibrations at 454 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e \u0026amp; 571 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e [24]. It is clear that the vibration peaks of Ni-O, Zn-O, \u0026amp; Cu-O that appear in the region below 600 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e overlap.\u003c/p\u003e\u003cp\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\u003eFTIR peaks with the functional groups of the synthesized CuO-NiO-ZnO NCs annealed at 200\u003csup\u003e\u0026ordm;\u003c/sup\u003eC \u0026amp; 500\u0026ordm;C.\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\"\u003e\u003cp\u003eCompounds\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePeak position (cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNature\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eType of Molecular motion\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e\u003cp\u003e\u003cb\u003eCuO-NiO-ZnO (200\u0026deg;C)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3462 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eStrong broad\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eO-H stretching\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2286 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eWeak broad\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eC\u0026thinsp;=\u0026thinsp;N stretching\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2048 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMedium\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eC\u0026thinsp;=\u0026thinsp;C\u0026thinsp;=\u0026thinsp;C stretching\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1983 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003cp\u003e1723 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003cp\u003e1351 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003cp\u003e1140 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eMedium\u003c/p\u003e\u003cp\u003eStrong\u003c/p\u003e\u003cp\u003eMedium\u003c/p\u003e\u003cp\u003eStrong\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eC\u0026thinsp;=\u0026thinsp;C\u0026thinsp;=\u0026thinsp;C stretching\u003c/p\u003e\u003cp\u003eC\u0026thinsp;=\u0026thinsp;O stretching\u003c/p\u003e\u003cp\u003eO-H Bending\u003c/p\u003e\u003cp\u003eC-O stretching\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e\u003cp\u003e\u003cb\u003eCuO-NiO-ZnO\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e(500\u0026deg;C)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1665 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eStrong broad\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eC\u0026thinsp;=\u0026thinsp;O stretching\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1620 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eStrong\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eC\u0026thinsp;=\u0026thinsp;C stretching\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1430 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003emedium\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eO-H stretching\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1142 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003emedium\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eC-O stretching\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"4. Antimicrobial Application","content":"\u003cp\u003eNanocomposites of tri-metallic nature have shown stronger antibacterial property, as compared to nanoparticles of mono-metallic \u0026amp; bi-metallic nature. The previous study on Au-Pt-Ag nanocomposite synthesized by green method which show effective antibacterial property against the bacteria types, (S. aureus \u0026amp; E. coli) [32\u0026ndash;34].\u003c/p\u003e\u003cp\u003eThe synthesized CuO/NiO/ ZnO nanocomposite shows antimicrobial action against the GNB (\u003cem\u003ePseudomonas\u003c/em\u003e \u0026amp; \u003cem\u003eE. coli\u003c/em\u003e) \u0026amp; GPB \u003cem\u003e(Bacillus)\u003c/em\u003e. These nanocomposites work against bacteria through different routes. The one of the important route is by reactive oxygen species (ROS) formation. These reactive species then disrupt the cycle of respiratory system, the cycle of protein transfer system, the food metabolism cycle, and DNA replication, causing cell death [35].\u003c/p\u003e\u003cp\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\u003eTabulation showing ZOI of GPB (Bacillus) \u0026amp; GNB \u003cem\u003e(Pseudomonas \u0026amp; E. coli)\u003c/em\u003e with the action of nanocomposites which are annealed at 200\u003csup\u003eO\u003c/sup\u003eC.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" 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=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSr. No\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eConcentration (ppm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eZOI\u003c/p\u003e\u003cp\u003e\u003cem\u003eBacillus\u003c/em\u003e\u003c/p\u003e\u003cp\u003e(GPB)\u003c/p\u003e\u003cp\u003e(mm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eZOI\u003c/p\u003e\u003cp\u003e\u003cem\u003ePseudomonas\u003c/em\u003e (GNB) (mm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eZOI\u003c/p\u003e\u003cp\u003e\u003cem\u003eE.coli\u003c/em\u003e\u003c/p\u003e\u003cp\u003e(GNB) (mm)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e40\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e5.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e6.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e250\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e7.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e500\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e17\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 antibacterial properties of the synthesized mixed MO nanocomposite were examined at several concentrations (5, 10, 20, 40, 50, 250, and 500 ppm). With an increase in concentration, the nanocomposite's antibacterial efficacy against two distinct bacterial strains improved. This could be the result of more reactive species that can stop the development of bacteria [36]. Figure\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003e displays pictures of the antibacterial properties of the synthesized mixed MO nanocomposite at different annealing temperatures of 200 \u0026amp; 500\u0026deg;C, respectively. The produced nanocomposite had outstanding antibacterial activity, according to the data. At a dose of 500 ppm, the largest ZOI against the GPB was seen\u003c/p\u003e\u003cp\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\u003eTabulation showing ZOI of GPB (Bacillus) \u0026amp; GNB \u003cem\u003e(Pseudomonas \u0026amp; E. coli)\u003c/em\u003e with the action of nanocomposites which are annealed at 500\u003csup\u003eO\u003c/sup\u003eC.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" 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=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSr. No\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eConcentration (ppm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eZOI\u003c/p\u003e\u003cp\u003e\u003cem\u003eBacillus\u003c/em\u003e\u003c/p\u003e\u003cp\u003e(GPB)\u003c/p\u003e\u003cp\u003e(mm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eZOI\u003c/p\u003e\u003cp\u003e\u003cem\u003ePseudomonas\u003c/em\u003e (GNB) (mm)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eZOI\u003c/p\u003e\u003cp\u003e\u003cem\u003eE.coli\u003c/em\u003e\u003c/p\u003e\u003cp\u003e(GNB) (mm)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e2.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e3.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e40\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e5.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e6.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e250\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e7.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e500\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e17\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\u003e\u003cb\u003eAction of Nanocomposites on Bacteria Cell.\u003c/b\u003e\u003c/p\u003e\u003cp\u003eIn addition to being crucial for sustaining bacterial growth and reproduction, typical bacterial metabolic activities can also result in illness. Bacterial cell death results from oxidative stress \u0026amp; damage to the cell membrane of bacterial cells caused by disruptions in bacterial metabolism. The effects of nanomaterials on bacterial metabolism have been explained by a variety of different processes, such as metal ion dissolution and reactive oxygen [60, 61]. Bacterial metabolism is a crucial biofilm function: The development and expansion of S. mutans biofilm depend on d-alanine metabolism [62].The production of ROS, which can damage membranes, DNA, \u0026amp; proteins, directly interacts with the cell membrane due to the ability of certain metal-based Nanocomposites to generate metal ions through dissolution, such as by inhibiting the chain of electron transport, \u0026amp; control of metabolic processes of bacterial cells are some of the ways that Nanocomposites can attack bacterial cells. The attack of Nanocomposites on bacterial cells through numerous mechanisms, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe results observed from the comparison between the antibacterial property of the ZnO-NiO-CuO NC examined in this work and previously published data, as observed in Table\u0026nbsp;\u003cspan refid=\"Tab8\" class=\"InternalRef\"\u003e5\u003c/span\u003e (a, b, c) below.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eTable 5 (a).\u003c/strong\u003e Comparison of antibacterial application by various nanocomposites for gram-positive (\u003cem\u003eBacillus\u003c/em\u003e) from the data obtained from earlier reported Studies.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"569\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSr. No\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMethod of \u0026nbsp;Synthesis\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZOI diameter (mm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 89px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eReferences\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003eNi-O\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003egreen synthesis (Stevia leaf extract)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 89px;\"\u003e\n \u003cp\u003e[51]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003eCu-O\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003egreen synthesis (Aerva javanica extract)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 89px;\"\u003e\n \u003cp\u003e[52]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003eZn-O\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003egreen synthesis (Cassia fistula extract)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 89px;\"\u003e\n \u003cp\u003e[53]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003eZnO-CuO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003egreen synthesis (Mentha longifolia leaf extract)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 89px;\"\u003e\n \u003cp\u003e[54]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003eCdO-NiO-ZnO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003emicrowave-assisted method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 89px;\"\u003e\n \u003cp\u003e[55]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 48px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003eCuO-NiO-ZnO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 210px;\"\u003e\n \u003cp\u003eCo-precipitation method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 89px;\"\u003e\n \u003cp\u003epresent study\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5 (b).\u003c/strong\u003e Comparison of Antibacterial Application by various Nanocomposites for \u003cstrong\u003eGram-negative bacteria\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003e(E. coli)\u0026nbsp;\u003c/em\u003e\u003c/strong\u003efrom the Data obtained from earlier Reported Studies.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"622\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSr. No\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 174px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMethod of \u0026nbsp;Synthesis\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZOI diameter (mm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eReferences\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003eNiO.CeO\u003csub\u003e2\u003c/sub\u003e. ZnO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 174px;\"\u003e\n \u003cp\u003eCo-precipitation method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e[56]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003eAg\u003csub\u003e2\u003c/sub\u003eO. CeO\u003csub\u003e2\u003c/sub\u003e. ZnO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 174px;\"\u003e\n \u003cp\u003eCo-precipitation method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e[57]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003eCdS-ZnO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 174px;\"\u003e\n \u003cp\u003eWet chemical synthesis\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e[58]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003eZnO:Cu/graphene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 174px;\"\u003e\n \u003cp\u003ePyrolysis method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e[59]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003eCdO\u0026minus;ZnO\u0026minus;NiO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 174px;\"\u003e\n \u003cp\u003eMicrowave-assisted method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003e[55]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 58px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003eCuO\u0026minus;ZnO\u0026minus;NiO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 174px;\"\u003e\n \u003cp\u003eCo-precipitation method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 115px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 127px;\"\u003e\n \u003cp\u003epresent study\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5(c).\u003c/strong\u003e Comparison of Antibacterial Application by various nanocomposites for Gram-negative bacteria \u003cem\u003e(Pseudomonas)\u0026nbsp;\u003c/em\u003efrom the data obtained from earlier reported Studies.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"626\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSr. No.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 183px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 168px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMethod of \u0026nbsp;Synthesis\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZOI diameter (mm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eReferences\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 183px;\"\u003e\n \u003cp\u003eNiO.CeO\u003csub\u003e2\u003c/sub\u003e. ZnO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 168px;\"\u003e\n \u003cp\u003eCo-precipitation method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e[56]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 183px;\"\u003e\n \u003cp\u003eAg\u003csub\u003e2\u003c/sub\u003eO. CeO\u003csub\u003e2\u003c/sub\u003e. ZnO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 168px;\"\u003e\n \u003cp\u003eCo-precipitation method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003e[57]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 49px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 183px;\"\u003e\n \u003cp\u003eCuO\u0026minus;ZnO\u0026minus;NiO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 168px;\"\u003e\n \u003cp\u003eCo-precipitation method\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 105px;\"\u003e\n \u003cp\u003epresent study\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\u003c/br\u003e\u003cp\u003eIt demonstrates that the produced nanocomposite has stronger antibacterial application as compared to single \u0026amp; bi-metallic nanoparticles that have been previously described. Furthermore, the recently described NCs can be examined for both antioxidant \u0026amp; antibacterial properties, while the CuO-NiO-ZnO NCs, which were previously published, were only examined for antimicrobial activity [37\u0026ndash;43].\u003c/p\u003e\u003cp\u003eAccording to Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003e, the synthesized nanocomposite exhibits superior antibacterial action against \u003cb\u003eBacillus\u003c/b\u003e (GPB) \u0026amp; \u003cb\u003ePseudomonas\u003c/b\u003e \u003cb\u003e\u0026amp;\u003c/b\u003e \u003cb\u003eE. coli\u003c/b\u003e (GNB). Because of the presence of the thick coating of peptidoglycan in GPB, it is simpler to target \u0026amp; harm the cell wall of the bacterial cell. GNB has an overabundance of lipopolysaccharides in its outer layer. Consequently, strains are more expensive than GPB. Consequently, it is discovered that the combined metal oxide CuO/NiO/ZnO, which is annealed at 500\u0026deg;C, has a stronger inhibitory effect on gram-positive bacteria [44\u0026ndash;50]. On the other hand, the combined MO CuO-NiO-ZnO, which is annealed at 200\u0026deg;C, has a stronger inhibitory effect on gram-negative bacteria.\u003c/p\u003e\u003cp\u003eGPB \u0026amp; GNB utilize distinct strategies to let the CuO-NiO-ZnO NCs into their cells; this might be due to variations in the composition of their membranes. The composition of the membrane determines how the CuO-NiO-ZnO NCs attach \u0026amp; are transported into the cell membrane of the bacterial cell.\u003c/p\u003e"},{"header":"5. Conclusions","content":"\u003cp\u003eThe chemical synthesis approach is one of the most efficient, economical, and time-saving approaches to developing the CuO-NiO-ZnO NCs. A higher concentration of the CuO-ZnO-NiO NCs proved to enhance its antibacterial activities. This might be because there are more particles accessible to interact with bacterial species. \u003cem\u003eBacillus\u003c/em\u003e (GPB) was more successfully inhibited by synthesized CuO-NiO-ZnO NCs annealed at 500\u0026deg;C than \u003cem\u003ePseudomonas\u003c/em\u003e \u0026amp; \u003cem\u003eE. coli\u003c/em\u003e (GNB). On the other hand, \u003cem\u003ePseudomonas\u003c/em\u003e \u0026amp; \u003cem\u003eE. coli\u003c/em\u003e (GNB) were more successfully inhibited by the synthesized CuO-NiO-ZnO NCs annealed at 200\u0026deg;C than \u003cem\u003eBacillus\u003c/em\u003e (GPB). The action of CuO-ZnO-NiO NCs upon the two species of bacteria differs because of variations in surface charge and cell wall composition. Using several characterization methods, including SEM, FTIR, XRD, \u0026amp; UV-visible spectroscopy, the physicochemical parameters of the samples were investigated. The study has investigated the application of nanocomposites for the development of antibacterial agents and coatings, which offers a gap of energy by nanocomposites of 2.70 eV \u0026amp; 2.67 eV at different annealing temperatures of 200\u0026deg;C and 500\u0026deg;C, respectively. Sharp \u0026amp; strong peaks observed in the XRD pattern confirmed the extremely crystalline structure of nanocomposites. The SEM micrograph confirms the heavily agglomerated background, which has visible voids at particular points. The study has investigated the application of nanocomposites for the development of antibacterial agents \u0026amp; coatings, which offer innovative strategies to combat infections.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eMO\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eMetal oxide\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eNCs\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eNanocomposites\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eNMs\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eNanomaterials\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eSEM\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eScanning electron microscopy\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eDLS\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eDynamic light scattering\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eXRD\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eX-ray diffraction\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eFTIR\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eFourier-transform infrared spectroscopy\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eUV\u0026ndash;Vis\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eUltraviolet\u0026ndash;visible spectroscopy\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eROS\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eReactive oxygen species\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eCB\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eConduction band\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003cdiv class=\"DefinitionListEntry\"\u003e\u003cdiv class=\"Term\"\u003eVB\u003c/div\u003e\u003cdiv class=\"Description\"\u003e\u003cp\u003eValence band\u003c/p\u003e\u003c/div\u003e\u003c/div\u003e\u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study involved only laboratory bacterial strains; no experiments on humans or animals were performed. According to the journal policies, formal human or animal ethics approval was not required. Informed consent was obtained from all individual participants included in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author confirms that informed consent for publication of the manuscript was obtained from all participants involved in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthorship contribution\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eVikas Chaudhary: Writing original draft, experimental, data curation, Kusham Lata- Writing review and editing, investigation, data curation, Manish Kumar: formal analysis, visualization, Ajay Sharma: formal analysis, visualization, Raman Kumar- formal analysis, Vivek Sheel Jaswal- Supervision, conceptualization, writing, review and editing, formal analysis, investigation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific grant from funding agencies.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo datasets were generated or analysed during the current study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eI am Kusham Lata, and I am highly thankful to DST-INSPIRE for providing a funding grant for my research work. We are also grateful to Central University of Himachal Pradesh for providing me with a platform for research work.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eJ. A. Rodriguez, X. Wang, J. C. Hanson, G. Liu, A. Iglesias-Juez, \u0026amp; M. Fern\u0026aacute;ndez-Garci´a. \u003cem\u003eThe behavior of mixed-metal oxides: Structural and electronic properties of Ce1\u0026minus;xCaxO2 and Ce1\u0026minus;xCaxO2\u0026minus;x. The Journal of Chemical Physics,\u003c/em\u003e\u003cstrong\u003e (2003)\u003c/strong\u003e\u003cem\u003e 119(11), 5659\u0026ndash;5669.\u003c/em\u003e https://doi.org/10.1063/1.1601595.\u003c/li\u003e\n\u003cli\u003eX. Jiang, X. Zhao, L. Duan, H. Shen, H. Liu, T. Hou, and F. Wang. Enhanced photoluminescence and photocatalytic activity of ZnO-ZnWO4 nanocomposites synthesized by a precipitation method. \u003cem\u003eCeramics International\u003c/em\u003e \u003cstrong\u003e(2016)\u003c/strong\u003e 42(14) 15160-15165. https://doi.org/10.1016/j.ceramint.2016.05.098.\u003c/li\u003e\n\u003cli\u003eZ. Wang, M. Yang, G. Shen, H. Liu, Y. Chen, and Q. Wang. Catalytic removal of benzene over CeO2\u0026ndash;MnO x composite oxides with rod-like morphology supporting PdO. \u003cem\u003eJournal of nanoparticle research\u003c/em\u003e \u003cstrong\u003e(2014)\u003c/strong\u003e 16(5) 2367. https://doi.org/10.1007/s11051-014-2367-5.\u003c/li\u003e\n\u003cli\u003eR.K. Sharma, R. Ghose. Synthesis of nanocrystalline CuO-ZnO mixed metal oxide powder by a homogeneous precipitation method, Ceram. Int. \u003cstrong\u003e(2014)\u003c/strong\u003e 40(7) 10919\u0026ndash;10926. https://doi.org/10.1016/j.ceramint.2014.03.089\u003c/li\u003e\n\u003cli\u003eP.P. Dorneanu, A. Airinei, N. Olaru, M. Homocianu, V. Nica, F. Doroftei, Preparation and characterization of NiO, ZnO and NiO\u0026ndash;ZnO composite nanofibers by electrospinning method, Mater. Chem. Phys. \u003cstrong\u003e(2014)\u003c/strong\u003e 148(3) 1029\u0026ndash;1035. https://doi.org/10.1016/j.matchemphys.2014.09.014\u003c/li\u003e\n\u003cli\u003eJ. Vidic, S. Stankic, F. Haque, D. Ciric, R. Le Goffic, A. Vidy, J. Jupille, B. Delmas, Selective antibacterial effects of mixed ZnMgO nanoparticles, J. Nanoparticle Res. 15(5). 1595. https://doi.org/10.1007/s11051-013-1595-4\u003c/li\u003e\n\u003cli\u003eG. Schimanke, M. Martin, J. Kunert, H. Vogel, Characterization of Mo-V-W Mixed Oxide Catalysts by ex situ and in situ X-Ray Absorption Spectroscopy, Z. Anorg. Allg. Chem. \u003cstrong\u003e(2005)\u003c/strong\u003e 631(6-7) 1289\u0026ndash;1296. https://doi.org/10.1002/zaac.200500064\u003c/li\u003e\n\u003cli\u003eH.R. Arandiyan, M. Parvari, Studies on mixed metal oxides solid solutions as heterogeneous catalysts, Brazilian J. Chem. Eng.\u003cstrong\u003e (2009)\u003c/strong\u003e 26 63\u0026ndash;74. https://doi.org/10.1590/S0104-66322009000100007\u003c/li\u003e\n\u003cli\u003eX. Li, R. Zhao, H. Jiang, Y. Zhai, P. Ma, Preparation and Catalytic Properties of ZnOCeO 2 -TiO 2 Composite, Synth. React. Inorganic, Met. Nano-Metal Chem.\u003cstrong\u003e (2016)\u003c/strong\u003e 46(5) 775\u0026ndash;782. https://doi.org/10.1080/15533174.2014.988806\u003c/li\u003e\n\u003cli\u003eH. R. Arandiyan, M. Parvari. Studies on Mixed Metal Oxides Solid Solutions as Heterogeneous Catalysts. \u003cem\u003eBraz. J. Chem. Eng.\u003c/em\u003e (\u003cstrong\u003e2009)\u003c/strong\u003e, \u003cem\u003e26\u003c/em\u003e (1), 63\u0026ndash;74. https://doi.org/10.1590/S0104-66322009000100007\u003c/li\u003e\n\u003cli\u003eC. Gionco, M. C. Paganini, S. Agnoli, A. E. Reeder, E. Giamello. Structural and Spectroscopic Characterization of CeO2\u0026ndash;TiO2 Mixed Oxides. \u003cem\u003eJ. Mater. Chem. A\u003c/em\u003e (\u003cstrong\u003e2013)\u003c/strong\u003e, \u003cem\u003e1\u003c/em\u003e(36), 10918. https://doi.org/10.1039/C3TA12018J\u003c/li\u003e\n\u003cli\u003eT. Dodevska, D. Hadzhiev, I. Shterev, and Y. Lazarova. Application of biosynthesized metal nanoparticles in electrochemical sensors. \u003cem\u003eJournal of the Serbian Chemical Society\u003c/em\u003e \u003cstrong\u003e(2022) \u003c/strong\u003e87(4) 401-435. https://doi.org/10.2298/JSC200521077D\u003c/li\u003e\n\u003cli\u003eA.K. Srivastava, S. Thota, J. Kumar, Preparation, microstructure and optical absorbtion behavior of NiO thin films, J. Nanosci. Nanotechnol.\u003cstrong\u003e (2008)\u003c/strong\u003e 8(8) 4111\u0026ndash;4115. https://doi.org/10.1166/jnn.2008.AN36\u003c/li\u003e\n\u003cli\u003eS.M. Abbas, S.T. Hussain. S. ali, N. Ahmad, N. Ali, S. Abbas, Z. Ali, Modification of carbon nanotubes by CuO-doped NiO nanocomposite for use as an anode material for lithium ion batteries, J. Sol. State. Chem.\u003cstrong\u003e (2013)\u003c/strong\u003e 202 43\u0026ndash;50. https://doi.org/10.1016/j.jssc.2013.03.036\u003c/li\u003e\n\u003cli\u003eM. Ben Amor, A. Boukhachem, A. Labidi, K. Boubaker, M. Amlouk, Physical investigations on Cd doped NiO thin films along with ethanol sensing at relatively low temperature, J. Alloys Compnd.\u003cstrong\u003e (2017)\u003c/strong\u003e 693 490\u0026ndash;499. https://doi.org/10.1016/j.jallcom.2016.09.207\u003c/li\u003e\n\u003cli\u003eA. Ezhilarasi, J. Judith Vijaya, K. Kaviyarasu, M. Maaza, A. Ayeshamariam, L. John Kennedy, Green synthesis of NiO nanoparticles using moringo oleifera extract and their nanoparticles biomedical applications: Cytotoxicity effect of nanoparticles against Ht29 cancer cells, J. Photochem. Photobiol. B: Biology \u003cstrong\u003e(2016)\u003c/strong\u003e 164 352\u0026ndash;360. https://doi.org/10.1016/j.jphotobiol.2016.10.003\u003c/li\u003e\n\u003cli\u003eA.E. Nel, L. Madler, D. Velegol, T. Xia, E.M.V. Hoek, P. Somasundaram, F. Klaessig, V. Castranova, M. Thompson, Nat. Mater \u003cstrong\u003e(2009)\u003c/strong\u003e 8(7):543-57. https://doi.org/10.1038/nmat2442\u003c/li\u003e\n\u003cli\u003e, A. El-Trass, H. ElShamy, I. El-Mehasseb, M. El-Kemary. CuO Nanoparticles: Synthesis, Characterization, Optical Properties and Interaction with Amino Acids. \u003cem\u003eApplied Surface Science (\u003c/em\u003e\u003cstrong\u003e2012)\u003c/strong\u003e, \u003cem\u003e258 \u003c/em\u003e(7), 2997\u0026ndash;3001. https://doi.org/10.1016/j.apsusc.2011.11.025\u003c/li\u003e\n\u003cli\u003eM. El-Kemary, N. Nagy, and I. El-Mehasseb. Nickel oxide nanoparticles: Synthesis and spectral studies of interactions with glucose. \u003cem\u003eMaterials Science in Semiconductor Processing\u003c/em\u003e 16(6) 1747-1752. https://doi.org/10.1016/j.mssp.2013.05.018\u003c/li\u003e\n\u003cli\u003eM. Pudukudy, Z. Yaakob. Facile Synthesis of Quasi Spherical ZnO Nanoparticles with Excellent Photocatalytic Activity. \u003cem\u003eJ Clust Sci (\u003c/em\u003e\u003cstrong\u003e2015)\u003c/strong\u003e, \u003cem\u003e26 \u003c/em\u003e(4), 1187\u0026ndash;1201. https://doi.org/10.1007/s10876-014-0806-1\u003c/li\u003e\n\u003cli\u003eK. Borgohain, J. Singh, M. Rama Rao, T. Shripathi, S. Mahamuni, Quantum size effects in CuO nanoparticles, Phys. Rev. B. \u003cstrong\u003e(2000)\u003c/strong\u003e 61 11093\u0026ndash;11096. https://doi.org/10.1103/PhysRevB.61.11093.\u003c/li\u003e\n\u003cli\u003eT. Ivanova, A. Harizanova, T. Koutzarova, B. Vertruyen, Study of ZnO sol-gel films: Effect of annealing, Mater. Lett.\u003cstrong\u003e (2010)\u003c/strong\u003e 64 1147\u0026ndash;1149. https://doi.org/10.1016/j.matlet.2010.02.033\u003c/li\u003e\n\u003cli\u003eZ. Yang, Z. Ye, Z. Xu, B. zhao, Effect of the morphology on the optical properties of ZnO nanostructures, Phys. E Low-Dimensional Syst. Nanostructures.\u003cstrong\u003e (2009)\u003c/strong\u003e 42 116\u0026ndash; 119. https://doi.org/10.1016/j.physe.2009.09.010\u003c/li\u003e\n\u003cli\u003eK. Anandan, V. Rajendran, Morphological and size effects of NiO nanoparticles via solvothermal process and their optical properties, Mater. Sci. Semicond. Process.\u003cstrong\u003e (2011)\u003c/strong\u003e 14 43\u0026ndash;47. https://doi.org/10.1016/j.mssp.2011.01.001\u003c/li\u003e\n\u003cli\u003eW.-Q. Wu, B.-X. Lei, H.-S. Rao, Y.-F. Xu, Y.-F. Wang, C.-Y. Su, D.-B. Kuang. Hydrothermal Fabrication of Hierarchically Anatase TiO2 Nanowire Arrays on FTO Glass for Dye-Sensitized Solar Cells. \u003cem\u003eSci Rep (\u003c/em\u003e\u003cstrong\u003e2013)\u003c/strong\u003e, \u003cem\u003e3 \u003c/em\u003e(1), 1352. https://doi.org/10.1038/srep01352\u003c/li\u003e\n\u003cli\u003eJuma, A. O.; Arbab, E. A. A.; Muiva, C. M.; Lepodise, L. M.; Mola, G. T. Synthesis and Characterization of CuO-NiO-ZnO Mixed Metal Oxide Nanocomposite. \u003cem\u003eJournal of Alloys and Compounds \u003c/em\u003e\u003cstrong\u003e2017\u003c/strong\u003e, \u003cem\u003e723\u003c/em\u003e, 866\u0026ndash;872. https://doi.org/10.1016/j.jallcom.2017.06.288\u003c/li\u003e\n\u003cli\u003eM.A. Subhan, T. Ahmed, N. Uddin, Synthesis, structure, PL and photocatalytic activities of La2O2CO3.CeO2.ZnO fabricated by coprecipitation method, Spectrochimic. Acta Part A: Mol. Biomol. Spectrosc \u003cstrong\u003e(2015)\u003c/strong\u003e 138, 827-833. https://doi.org/10.1016/j.saa.2014.10.114\u003c/li\u003e\n\u003cli\u003eM.A. Subhan, T. Ahmed, M.R. Awal, A.M.M. Fahim, Synthesis, structure and excitation wavelength dependent PL properties of novel nanocomposite La2O2CO3.CuO.ZnO, Spectrochimic. Acta Part A: Mol. Biomol. Spectrosc \u003cstrong\u003e(2014)\u003c/strong\u003e 132, 550-554 https://doi.org/10.1016/j.saa.2014.05.015\u003c/li\u003e\n\u003cli\u003eM.A. Subhan, T. Ahmed, Synthesis, characterization and spectroscopy investigations of novel nano multi-metal oxide Co3O4. CeO2.ZnO, Spectrochimic. Acta Part A: Mol. Biomol. Spectrosc \u003cstrong\u003e(2014) \u003c/strong\u003e129, 377-381. https://doi.org/10.1016/j.saa.2014.03.051\u003c/li\u003e\n\u003cli\u003eM.A. Subhan, T. Ahmed, N. Uddin, A.K. Azad, K. Begum, Synthesis, characterization, PL properties, photocatalytic and antibacterial activities of nano multi-metal oxide NiO.CeO2.ZnO, Spectrochimic. Acta Part A. Mol. Biomol. Spectrosc \u003cstrong\u003e(2015) \u003c/strong\u003e136, 824-831. https://doi.org/10.1016/j.saa.2014.09.100\u003c/li\u003e\n\u003cli\u003eK. Karthik, S. Dhanuskodi, C. Gobinath\u003cem\u003e.\u003c/em\u003e Multifunctional properties of microwave assisted CdO\u0026ndash;NiO\u0026ndash;ZnO mixed metal oxide nanocomposite: enhanced photocatalytic and antibacterial activities. \u003cem\u003eJ Mater Sci: Mater Electron\u003c/em\u003e \u003cstrong\u003e(2018) 29(7)\u003c/strong\u003e, 5459\u0026ndash;5471. https://doi.org/10.1007/s10854-017-8513-y\u003c/li\u003e\n\u003cli\u003eM. Hafeez, R. Arshad, J. Khan, B. Akram, M. N. Ahmad, M. U. Hameed, S. Haq. Populus ciliata mediated synthesis of copper oxide nanoparticles for potential biological applications. Mater. Res. Express. \u003cstrong\u003e(2019) \u003c/strong\u003e6(5), 055-043. https://doi.org/10.1088/2053-1591/ab0601\u003c/li\u003e\n\u003cli\u003eD. Paul, S. Mangla, S. Neogi. Antibacterial study of CuO-NiO-ZnO trimetallic oxide nanoparticle. Mater. Lett. \u003cstrong\u003e(2020)\u003c/strong\u003e, 271, 127740. https://doi.org/10.1016/j.matlet.2020.127740\u003c/li\u003e\n\u003cli\u003eL. Tao, D. Dastan, W. Wang, P. Poldorn, X. Meng, M. Wu, H. Zhao, H. Zhang, L. Li, B . An. Metal-Decorated InN Monolayer Senses N2 against CO2. ACS Appl. Mater. Interfaces \u003cstrong\u003e(2023),\u003c/strong\u003e 15, 12534\u0026minus; 12544. https://doi.org/10.1021/acsami.2c21463\u003c/li\u003e\n\u003cli\u003eD. S. Ahmed, M.K.A. Mohammed. Studying the bactericidal ability and biocompatibility of gold and gold oxide nanoparticles decorating on multi-wall carbon nanotubes. Chem. Papers \u003cstrong\u003e(2020),\u003c/strong\u003e 74, 4033\u0026minus;4046. https://doi.org/10.1007/s11696-020-01223-0\u003c/li\u003e\n\u003cli\u003eB. Luan, C. Liu, P. Chen, and R. Chen. \u0026quot;Nanotechnology Reviews \u0026ldquo;Just Accepted\u0026rdquo; Papers Nanotechnol.\u0026quot; (2011). \u003c/li\u003e\n\u003cli\u003eM. KA. Mohammed, Ali K. Al-Mousoi, S. M. Majeed, S. Singh, A. Kumar, R. Pandey, J. Madan, D. S. Ahmed, and D. Dastan. Stable hole-transporting material-free perovskite solar cells with efficiency exceeding 14% via the introduction of a malonic acid additive for a perovskite precursor. \u003cem\u003eEnergy \u0026amp; Fuels\u003c/em\u003e \u003cstrong\u003e(2022)\u003c/strong\u003e 36(21) 13187-13194. https://doi.org/10.1021/acs.energyfuels.2c02878\u003c/li\u003e\n\u003cli\u003eM. KA. Mohammed, Ali K. Al-Mousoi, S. M. Majeed, S. Singh, A. Kumar, R. Pandey, J. Madan, D. S. Ahmed, and D. Dastan. Ionic Liquid Passivator for Mesoporous Titanium Dioxide Electron Transport Layer to Enhance Efficiency and Stability of Hole Conductor-Free Perovskite Solar Cells. Energy Fuels \u003cstrong\u003e(2022),\u003c/strong\u003e 36(19), 12192\u0026minus;12200. https://doi.org/10.1021/acs.energyfuels.2c01980\u003c/li\u003e\n\u003cli\u003eB. Akram, K. Ahmad, J. Khan, B. A. Khan, and J. Akhtar. Low-temperature solution-phase route to sub-10 nm titanium oxide nanocrystals having super-enhanced photoreactivity. \u003cem\u003eNew Journal of Chemistry\u003c/em\u003e \u003cstrong\u003e(2018) \u003c/strong\u003e42(13) 10947-10952. https://doi.org/10.1039/C8NJ02317D\u003c/li\u003e\n\u003cli\u003eP. Zhu, D. Dastan, L. Liu, L. Wu, Z. Shi, Q.-Q. Chu, F. Altaf, and M. KA Mohammed. Surface wettability of various phases of titania thin films: Atomic-scale simulation studies. \u003cem\u003eJournal of Molecular Graphics and Modelling\u003c/em\u003e \u003cstrong\u003e(2023)\u003c/strong\u003e 118 108335. https://doi.org/10.1016/j.jmgm.2022.108335\u003c/li\u003e\n\u003cli\u003eI. Ashraf, S. Ahmad, F. Nazir, D. Dastan, Z. Shi, H. Garmestani, and M. Iqbal. Hydrothermal synthesis and water splitting application of d-Ti3C2 MXene/V2O5 hybrid nanostructures as an efficient bifunctional catalyst. \u003cem\u003eInternational Journal of Hydrogen Energy\u003c/em\u003e \u003cstrong\u003e(2022)\u003c/strong\u003e 47(64) 27383-27396. https://doi.org/10.1016/j.ijhydene.2022.06.095\u003c/li\u003e\n\u003cli\u003eJ. Dlugaszewska, R. Dobrucka. Effectiveness of biosynthesized trimetallic Au/Pt/Ag nanoparticles on planktonic and biofilm Enterococcus faecalis and Enterococcus faecium forms. J. Clust. Sci. \u003cstrong\u003e(2019),\u003c/strong\u003e 30, 1091\u0026minus;1101. https://doi.org/10.1007/s10876-019-01570-3\u003c/li\u003e\n\u003cli\u003eG. L. Tan, D. Tang, D. Dastan, A. Jafari, Z. Shi, Q.-Q. Chu, J. PB Silva, and X.-T. Yin. Structures, morphological control, and antibacterial performance of tungsten oxide thin films. \u003cem\u003eCeramics International\u003c/em\u003e \u003cstrong\u003e(2021)\u003c/strong\u003e 47(12) 17153-17160. https://doi.org/10.1016/j.ceramint.2021.03.025\u003c/li\u003e\n\u003cli\u003eR. Mohammadi-Aloucheh, A. Habibi-Yangjeh, A. Bayrami, S. Latifi-Navid, and A. Asadi. Green synthesis of ZnO and ZnO/CuO nanocomposites in Mentha longifolia leaf extract: characterization and their application as anti-bacterial agents. \u003cem\u003eJournal of Materials Science: Materials in Electronics\u003c/em\u003e \u003cstrong\u003e(2018)\u003c/strong\u003e 29(16) 13596-13605. https://doi.org/10.1007/s10854-018-9487-0\u003c/li\u003e\n\u003cli\u003eM. Nasrollahzadeh, M. Sajjadi, S. Iravani, and R. S. Varma. Trimetallic nanoparticles: greener synthesis and their applications. \u003cem\u003eNanomaterials\u003c/em\u003e \u003cstrong\u003e(2020)\u003c/strong\u003e 10(9) 1784. https://doi.org/10.3390/nano10091784\u003c/li\u003e\n\u003cli\u003eV. Manikandan, P. Velmurugan, J. H. Park, N. Lovanh, S. K. Seo, P. Jayanthi, Y. J. Park, M. Cho, and B. T. Oh. Synthesis and antimicrobial activity of palladium nanoparticles from Prunus\u0026times; yedoensis leaf extract. \u003cem\u003eMaterials Letters\u003c/em\u003e \u003cstrong\u003e(2016)\u003c/strong\u003e 185 335-338. https://doi.org/10.1016/j.matlet.2016.08.120\u003c/li\u003e\n\u003cli\u003eS. Srihasam, K. Thyagarajan, M. Korivi, V. R. Lebaka, and S. P. R. Mallem. Phytogenic generation of NiO nanoparticles using Stevia leaf extract and evaluation of their in-vitro antioxidant and antimicrobial properties. \u003cem\u003eBiomolecules\u003c/em\u003e \u003cstrong\u003e(2020)\u003c/strong\u003e 10(1) 89 https://doi.org/10.3390/biom10010089\u003c/li\u003e\n\u003cli\u003eF. Amin, B. Khattak, A. Alotaibi, M. Qasim, I. Ahmad, R. Ullah, M. Bourhia, A. Gul, S. Zahoor, R. Ahmad. Green synthesis of copper oxide nanoparticles using Aerva javanica leaf extract and their characterization and investigation of in vitro antimicrobial potential and cytotoxic activities. Evidence-Based Complement. Altern. Med. \u003cstrong\u003e(2021)\u003c/strong\u003e, 2021, 1\u0026minus;12. https://doi.org/10.1155/2021/5589703\u003c/li\u003e\n\u003cli\u003eD. Suresh, P. C. Nethravathi, H. Rajanaika, H. Nagabhushana, and S. C. Sharma. \u0026quot;Green synthesis of multifunctional zinc oxide (ZnO) nanoparticles using Cassia fistula plant extract and their photodegradative, antioxidant and antibacterial activities.\u0026quot; \u003cem\u003eMaterials Science in Semiconductor Processing\u003c/em\u003e \u003cstrong\u003e(2015)\u003c/strong\u003e 31 446-454. https://doi.org/10.1016/j.mssp.2014.12.023\u003c/li\u003e\n\u003cli\u003eF. Sowmiya, , S. R. Brintha. Synthesis and Characterization of Zno-Cdo Nanocomposite and Its Antibacterial Activity. J. Pharm. Negat. Results \u003cstrong\u003e(2022),\u003c/strong\u003e 13, 3559\u0026minus;3569.\u003c/li\u003e\n\u003cli\u003eW. Chen, L. Yue, Q. Jiang, X. Liu, and W. Xia.Synthesis of varisized chitosan-selenium nanocomposites through heating treatment and evaluation of their antioxidant properties. Int. J. Biol. Macromol. \u003cstrong\u003e(2018),\u003c/strong\u003e 114, 751\u0026minus;758. https://doi.org/10.1016/j.ijbiomac.2018.03.108\u003c/li\u003e\n\u003cli\u003eB.V. Vibitha , B. Anitha, P. G. A. Krishna, and J. N. Tharayil. Plant extracts assisted synthesis, characterization and antioxidant properties of ZnO: CuO nanocomposites. In \u003cem\u003eAIP Conference Proceedings\u003c/em\u003e, vol. 2244(1), p. 070037. AIP Publishing LLC, 2020. https://doi.org/10.1063/5.0009174\u003c/li\u003e\n\u003cli\u003eS. Hajji, R. B. S.-B. Salem, M. Hamdi, K. Jellouli, W. Ayadi, M. Nasri, and S. Boufi. Nanocomposite films based on chitosan\u0026ndash;poly (vinyl alcohol) and silver nanoparticles with high antibacterial and antioxidant activities. \u003cem\u003eProcess Safety and Environmental Protection\u003c/em\u003e \u003cstrong\u003e(2017) \u003c/strong\u003e111 112-121. https://doi.org/10.1016/j.psep.2017.06.018\u003c/li\u003e\n\u003cli\u003eP. Siriwong, T. Thongtem, A. Phuruangrat, and S. Thongtem. Hydrothermal synthesis, characterization, and optical properties of wolframite ZnWO 4 nanorods. \u003cem\u003eCrystEngComm\u003c/em\u003e \u003cstrong\u003e(2011):\u003c/strong\u003e 13(5) 1564-1569. https://doi.org/10.1039/C0CE00402B\u003c/li\u003e\n\u003cli\u003e[55 K. Karthik, S. Dhanuskodi, C. Gobinath\u003cem\u003e.\u003c/em\u003e Multifunctional properties of microwave assisted CdO\u0026ndash;NiO\u0026ndash;ZnO mixed metal oxide nanocomposite: enhanced photocatalytic and antibacterial activities. \u003cem\u003eJ Mater Sci: Mater Electron\u003c/em\u003e \u003cstrong\u003e(2018) 29(7)\u003c/strong\u003e, 5459\u0026ndash;5471. https://doi.org/10.1007/s10854-017-8513-y\u003c/li\u003e\n\u003cli\u003eM.A. Subhan, T. Ahmed, N. Uddin, A.K. Azad, K. Begum, Synthesis, characterization, PL properties, photocatalytic and antibacterial activities of nano multi-metal oxide NiO.CeO2.ZnO, Spectrochimic. Acta Part A. Mol. Biomol. Spectrosc \u003cstrong\u003e(2015) \u003c/strong\u003e136, 824-831. https://doi.org/10.1016/j.saa.2014.09.100\u003c/li\u003e\n\u003cli\u003eM.A. Subhan, N. Uddin, P. Sarker, H. Nakata, R. Makioka, Synthesis, characterization, low temperature solid state PL and photocatalytic activities of Ag2O, CeO2.ZnO nanocomposite, Spectrochimic. Acta Part A. Mol. Biomol. Spectrosc \u003cstrong\u003e(2015) \u003c/strong\u003e151, 56-63 https://doi.org/10.1016/j.saa.2015.06.094\u003c/li\u003e\n\u003cli\u003eT.K. Jana, S.K. Maji, A. Pal, R.P. Maiti, T.K. Dolai, K. Chatterjee, Photocatalytic and antibacterial activity of cadmium sulphide/zinc oxide nanocomposite with varied morphology. J. Colloid Interf. Sci \u003cstrong\u003e(2016) \u003c/strong\u003e480, 9-16 https://doi.org/10.1016/j.jcis.2016.06.073\u003c/li\u003e\n\u003cli\u003eK. Ravichandran, N. Chidhambaram, S. Gobalakrishnan, Copper and graphene activated ZnO nanoparticles for enhanced photocatalytic and antibacterial activities. J. Phys. Chem. Solids \u003cstrong\u003e(2016) \u003c/strong\u003e93, 82-90 https://doi.org/10.1016/j.jpcs.2016.02.013\u003c/li\u003e\n\u003cli\u003eN. Padmavathy, R. Vijayaraghavan . Interaction of ZnO nanoparticles with microbes \u0026ndash; a physio and biochemical assay. J Biomed Nanotechnol. \u003cstrong\u003e(2011)\u003c/strong\u003e 7(6) 813\u0026ndash;822 https://doi.org/10.1166/jbn.2011.1343\u003c/li\u003e\n\u003cli\u003eJ. Yu, W. Zhang, Y. Li. Synthesis, characterization, antimicrobial activity and mechanism of a novel hydroxyapatite whisker/nano zinc oxide biomateriali. Biomed Mater. \u003cstrong\u003e(2014)\u003c/strong\u003e 10(1):015001. https://doi.org/10.1088/1748-6041/10/1/015001.\u003c/li\u003e\n\u003cli\u003eW. Qiu, X. Zheng, Y. Wei. D-alanine metabolism is essential for growth and biofilm formation of Streptococcus mutans. Mol Oral Microbiol. \u003cstrong\u003e(2016)\u003c/strong\u003e 31(5), 435\u0026ndash;444. https://doi.org/10.1111/omi.12146\u003c/li\u003e\n\u003cli\u003eJ. Iqbal, B. A. Abbasi, A. Munir, S. Uddin, S. Kanwal, and T. Mahmood. Facile green synthesis approach for the production of chromium oxide nanoparticles and their different in vitro biological activities. \u003cem\u003eMicroscopy research and technique\u003c/em\u003e \u003cstrong\u003e(2020)\u003c/strong\u003e 83(6) 706-719. https://doi.org/10.1002/jemt.23460\u003c/li\u003e\n\u003cli\u003eJ. Stetefeld, S. A. McKenna, and T. R. Patel, Dynamic light scattering: a practical guide and applications in biomedical sciences. Biophysical reviews. \u003cstrong\u003e(2016)\u003c/strong\u003e 8(4) 409-27. https://doi.org/10.1007/s12551-016-0218-6\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"catalysis-letters","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Catalysis Letters](https://link.springer.com/journal/10562)","snPcode":"10562","submissionUrl":"https://submission.springernature.com/new-submission/10562/3","title":"Catalysis Letters","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"CuO-NiO-ZnO, nanocomposite, antibacterial, concentration","lastPublishedDoi":"10.21203/rs.3.rs-7532947/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7532947/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe synthesis, characterization, \u0026amp; use of mixed metal oxides, a unique family of compounds, have attracted ever-increasing interest as they offer an excellent opportunity to refine the desired properties for improved functional performance in interaction with base metal oxides. In this study, NiO-ZnO-CuO composites are synthesized by using simple co-precipitation technique at 200\u003csup\u003eo\u003c/sup\u003eC and 500\u003csup\u003eo\u003c/sup\u003eC temperature and then characterized for its properties by UV-visible, X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) studies, Dynamic Light Scattering (DLS) and Scanning electron microscopy (SEM) analysis. At 200°C \u0026amp; 500°C, the CuO-NiO-ZnO nanocomposite had an optical band gap energy of 2.70 eV \u0026amp; 2.67 eV, respectively. The UV-visible spectroscopy was utilized to find the band gap using the Tauc plot. XRD studies indicated the presence of well-defined CuO (monoclinic), NiO (cubic), \u0026amp; ZnO (hexagonal) phases, with the maximum fraction of volume belonging to the NiO. The images of SEM by mixed oxide nanocomposites display nanoparticles in the form of irregular shapes. The phases indicate the presence of trimetallic oxide heterojunctions, which strongly impact the overall properties of nanocomposites (NCs). Also, the antibacterial activities of nanocomposites are studied, which showed that at 200°C, the nanocomposite exhibited antibacterial action against gram-negative bacteria (GNB), whereas at 500°C, it demonstrated efficacy against gram-positive bacteria (GPB) and also affected GPB at higher doses.\u003c/p\u003e","manuscriptTitle":"Antibacterial Application of Heterogeneous CuO-NiO-ZnO Metal Oxides Nanocomposites","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-17 18:52:11","doi":"10.21203/rs.3.rs-7532947/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-09-16T14:47:17+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-16T12:10:43+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-09-11T12:32:49+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"147772660391377003611306022246456272627","date":"2025-09-10T15:41:51+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"33638056782066870931089371302663842456","date":"2025-09-10T12:32:11+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-10T09:15:03+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-08T03:33:36+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-08T03:32:31+00:00","index":"","fulltext":""},{"type":"submitted","content":"Catalysis Letters","date":"2025-09-04T06:46:56+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"catalysis-letters","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Catalysis Letters](https://link.springer.com/journal/10562)","snPcode":"10562","submissionUrl":"https://submission.springernature.com/new-submission/10562/3","title":"Catalysis Letters","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"4857570c-8333-40f7-97f9-6a51c3fe66e8","owner":[],"postedDate":"September 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2025-10-09T12:53:32+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-17 18:52:11","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7532947","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7532947","identity":"rs-7532947","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}