INTRODUCTION
The alarming growth of infectious diseases and dangerous microorganisms is a major cause for concern. Compared with their bulk equivalents, particles at the nanoscale exhibit unique physicochemical characteristics. A number of medical uses exist for nanoparticles (NPs), including biosensors, imaging, cancer treatment, drug delivery systems, diagnostic tools, and biosensors [2–4]. NPs possess a high surface-to-volume ratio, endowing them with distinctive features and enhancing their potential for biomedical applications. NPs are primarily categorized into three categories in biological applications on the basis of their chemical structure [5,6]. And have a bigger surface area available for contact with microorganisms than micro particles Organic (such as polymers and liposomes) and inorganic NPs (such as metals, metal oxides, quantum dots, ceramics, and carbon-based NPs) are encompassed by this phrase [7–9]. Various methodologies for synthesizing metallic NPs include chemical reduction, electrochemical synthesis, hydrothermal methods, electrophoresis, photochemical synthesis, template-assisted synthesis, chemical vapor deposition, sol–gel processes, pyrolysis, and nanoemulsion. The unique physicochemical properties, biocompatibility, and therapeutic potential of Zn NPs have garnered significant interest as novel nanomaterials. The scientific community is highly intrigued by the potential of Zn NPs as a cancer treatment. Several studies have confirmed that these particles are effective against cancer cells [11]. Zn NPs provide numerous advantageous characteristics and distinct benefits compared with other NP kinds in terms of efficacy, selectivity, and the development of medicine resistance. Ginger, the rhizome of Zingiber officinale, has been utilized for therapeutic purposes since ancient times. It has been a significant plant in the old pharmacopoeias of China and India. Various constituents of ginger, including 6-gingerol, 6-shogaol, and Galano lactone, have been documented to exhibit anti-5HT3 action [12,13]. Z. officinale is utilized to address a range of gastrointestinal issues, including motion sickness, morning sickness, dyspepsia, flatulence, and diarrhea. It is utilized for the management of nausea induced by cancer therapy, postoperative nausea and vomiting, and appetite loss [14–16]. It is also utilized as a digestive aid for mild gastrointestinal disturbances, cough, persistent bronchial issues, low-grade infections of various types, and anorexia situations. Z. officinale is presently available in the forms of extract (Zinaxin), tincture (Bioforce Jan De Vries), pill, and oil for oral use. The present study examined the impact of Zn and aqueous ginger extract, produced via pulsed laser ablation mothed, on certain physical attributes and biological activities of specific bacterial strains.

MATERIASL AND METHODS
Synthesis of Zn NPs
A pure zinc plate (purity level of 99.66%), measuring (1 cm × 1 cm × 2 mm), was immersed in 5 mL of deionized distilled water in a beaker 5 cm from the laser source. It was used in the pulsed laser ablation technique in liquid to remove organic contaminants from the target, which had been previously purified using ethanol and distilled water. A Q-switched Nd:YAG laser that pulses 1000 times per wavelength, has an energy output of 500 mJ, and operates at a repetition rate of 1 Hz was used at a wavelength of 1064 nm. First, plant extract was prepared by purifying and rinsing zingiber root leaves with deionized water. Subsequently, the plants were chopped into small pieces, added to a 100 mL beaker together with 80 mL of deionized water, and stirred at 80 °C for 1 h. Filter paper was used for filtering to further purify the extract solution. Fig. 1 shows the overall process for creating ZnNPs via pulsed laser ablation using zingiber extract. ZnNPs were identified, and their synthesis from ginger extracts was verified using absorption spectra and surface plasmon resonance (SPR) obtained using a Shimadzu Double Beam 1800 UV spectrometer from Japan. Nano-solutions were mixed with KBr granules to determine the effective aggregation of NPs synthesized in different solvents, and the infrared spectra were characterized across the wavenumber range of 400–4000 cm−1 by using a Shimadzu IR Affinity-1CE (Fourier transform infrared spectroscopy, FTIR) spectrophotometer from Japan. field emission scanning electron microscope (FESEM) model (ZEISS SIGMAVP/ Germany).

Antibacterial Activity Tests
 Staphylococcus aureus, Streptococcus spp., Escherichia coli, and Klebsiella pneumoniae were grown on blood agar. Biomeriex’s calibration solution was used to count the number of bacterial cells (about 1.5 × 108 cells/mL). With a loop, a series of bacterial colonies was moved to create suspended bacteria. These bacteria were then placed into tubes with brain heart infusion broth. The tubes were kept at 37° C for 18–24 h. The suspended bacteria were evaluated against the standard MacFarland solution at a concentration of 1.5 × 108 cells/ml. Afterwards, the suspended bacteria were placed on Mueller–Hinton agar plates with a sterile brush and left to dry for a short period. Holes were made in the culture material with a sterilized cork borer. A micropipette was used to place 100 µL of the medication into each well at the doses of 100, 75, 50, 25, and 12.5 µg/mL. The diameter of the inhibitory zone surrounding each hole was assessed to determine how effectively each concentration worked.

Hemolytic Toxicity by Red Blood Cell (RBC) Fragility Test
The RBC fragility test was used to see how well the chemicals could break down RBCs. Fresh human RBCs were washed three times with isotonic saline (0.9% NaCl) and then resuspended to a 2% hematocrit. An equal amount of erythrocyte suspension was mixed with chemical solutions and kept at 37 °C for 1 h. After centrifugation was performed at 1500 rpm for 10 min, the supernatant’s absorbance was measured at 540 nm to determine the amount of hemoglobin released. Distilled water was used to check for complete hemolysis (positive control), and isotonic saline was used as the negative control. The percentage of hemolysis was compared with the controls. Osmotic lysis frees hemoglobin from RBCs, and the amount of this protein in the supernatant is directly related to the number of cells that have been lysed. Photometric measurements at 540 nm were conducted to measure the hemoglobin concentration in the supernatants to show that hemolysis occurred. The absorbance of the supernatant from erythrocytes lysed in distilled water was established as 100% hemolysis. The following equation was used to determine how much hemolysis occurred:

In vitro hemolytic assay using spectrophotometry is a direct and useful means to measure hemolysis quantitatively. This approach assesses the impact of varying biomolecule concentrations on human RBCs. The evaluation of cytotoxicity by hemolytic activity assays has proven to be an efficient alternative screening tool for fundamental toxicity. The evaluation of erythrocyte hemolytic activity against various substances is rapid, reliable, and cost-effective, hence facilitating a reduction in the use of laboratory animals for in vivo testing and furthering the objectives of minimizing, refining, and replacing animal research. The hemolytic activity on human erythrocytes is determined to assess toxicity profiles for human application. The level of in vitro cytotoxicity associated with hemolytic activity is assessed on the basis of mortality rate: 0%–9% indicates non-toxicity, 10%–49% refers to minor toxicity; 50%–89% denotes toxicity, and 90%–100% reflects high toxicity. 

Antioxidant Activity by DPPH Free Radical Scavenging Assay
DPPH radical scavenging experiment was conducted to see how well the chemicals worked as antioxidants. A methanol solution of 0.1 mM DPPH was established. One milliliter of each compound was combined with 1 mL of DPPH solution. The mixtures were kept in the dark at room temperature for 30 min, and the amount of light absorbed at 517 nm was determined. Ascorbic acid served as the positive control. The following formula was used to identify the proportion of radical scavenging activity:

where A control is the absorbance of DPPH solution without sample, and sample A is the absorbance with the compound test.

RESULTS AND DISCUSSION
Many different substances, such as transition metal ions, organic compounds, and biological molecules, can be found using this method [19]. The first sign of nano production was the change in color of Zn colloidal solutions from clear to yellowish-brown, with the color changing depending on how many pulses were used. UV–Vis analysis was performed to obtain a first look at ZnNPs. Fig. 2 shows that the peak absorption range was between 295 and 297 nm when the Nd:YAG laser had a wavelength of 1064 nm and an energy of 500 mJ. The highest absorption showed the SPR of NPs. Fig. 2 shows how the hue of the synthesis solution changed. The Zn combination and the extract solution transitioned from colorless to pale yellow, with the color being constant post-reaction. UV–Vis spectra were acquired relative to water to validate the production of ZnNPs. Fig. 2 illustrates the UV–Vis spectra associated with the ZnNP creation utilizing ginger extracts through pulsed laser ablation. The signal that can be observed in the 400–460 nm region indicated the SPR of the synthesized ZnNPs [20]. The intensity of SPR increased with prolonged reaction time, indicating the production of more ZnNPs. Clusters of ZnNP were continuously generated.
The optical band gap (Eg) of the NPs was ascertained by extrapolating a plot of (αhv) versus the photon energy in the absorption spectra. The absorption coefficient (α) is expressed as (αhv)², and the intercept provides a dependable estimation of the band gap (Eg) [21]. Fig. 3 demonstrates that the energy band gaps were 4.25 eV for Zn, and 3.5 eV for Zn: Zingiber. The reduction in the band gap correlated with an increase in concentration (the specified eliminator duration), which can be attributed to the quality of the prepared sample. As the eliminator’s time increased, the resulting colloid exhibited a more pronounced yellow hue attributed to the ginger extract. The band gap indicates the transfer of electrons from the valence band to the conduction band. A diminished band gap value results in an enhanced transition of electrons to the excited state, hence influencing the adsorption intensity of NPs. The adsorption intensity indicates the transition activity from the excited state to the ground state. The integration of ginger extract into Zn NPs altered the band gap value. The ginger extract can substitute for one of the lattice sites in the Zn crystal structure. Zn has a reduced band gap energy value [22].
The FTIR spectra in Fig. 4 showed absorption peaks that were linked to the vibrations of hydroxyl (O-H), alkane (C-H), aliphatic amines (C-O), and a carboxylic acid (C-O) group. In both pictures, the band below 625 cm−1 showed that ZnNPs and their oxides formed. The number of pulses affected the strength of the absorption peaks and the presence or absence of certain peaks because the number of pulses changes the concentration of the solution [23]. The spectra at 3288 and 3275 cm−1 were likely due to the ambient moisture during the analysis, and they were related to the OH-stretch bonds of the carboxyl group. The peaks at 1321.2 and 1402.2 cm−1 can be attributed to C-O vibrational modes, indicating that CO2 dissolving in air has no effect [24]. FTIR examination of G-ZnO NPs showed that Z. officinale extract works as a reducing and capping agent. The broadband coupling at 3439 cm−1 showed that the broadband (OH) vibrated in a manner that makes it expand. Peaks were found at 2980, 1579, 1420, 1036, 630, and 465 cm−1, along with their wave numbers and functional groups. Fig. 4 shows organic compounds such as polyphenols, carboxylic and aromatic acids, terpenes, and phenols. The signal at 1579 cm−1 clearly showed aromatic compounds, and the peaks at 630 and 465 cm−1 showed that G-ZnO NPs were formed by stretching the Zn-O bond. The biological parts of Z. officinale are exactly copied on the surface of NPs [25,26].
Measurement (FE-SEM) of Zn nanoparticles synthesized via ginger extract synthesized by laser ablation was carried out after deposition on quartz as in Fig. 5, where the diameters of the nanoparticles and the standard deviation and largest and smallest diameters of the nanoparticles were recorded. According to scanning electron microscopy (SEM) analysis, the produced zinc oxide nanoparticles (ZnONPs) are spherical and agglomerated, in a previous study, Vidya et al. reported the synthesis of spherical nanoparticles ranging in diameter from 11 to 25 nm [27]. In our study, however, the diameters range from 45 to 78 nm.

Antibacterial Activity
Fig. 6 and Table 1 show how Zn NPs affects S. aureus, Streptococcus spp., E. coli, and K. pneumoniae and Fig. 6 shows Inhibition rate of type of bacteria by using several solutions of Zn NPs. This effect may be attributed to the different sizes and shapes of the Zn NPs made utilizing the green approach. It had an effect on germs, although not as much as its predecessor. The different sizes of NPs, which are very important, alter how they affect bacteria. A larger size of NPs makes it easier for them to go through the cell wall and into the nucleus [28]. NPs exhibit increased interaction with bacterial cell membranes, hence augmenting their toxicity in comparison to spherical NPs [29]. NPs effectively target pathogenic bacteria because their surface area is much larger than their volume. Smaller NPs have a larger surface area, which increases their surface charge and accelerates the process of killing or stopping bacteria through electrostatic attraction [28,30]. Fig. 7 shows Inhibition rate of type of bacteria by using several solutions of Zn NPs.

RBC Fragility (Hemolysis) Assay 
In the RBC fragility (hemolysis) experiment, the absorbance of the supernatant at around 540 nm quantitatively represents free hemoglobin; thus, a higher absorbance suggests increased membrane damage and RBC lysis. In this work, pure water functioned as the positive control (100% hemolysis) and isotonic saline served as the negative control (0% hemolysis) in accordance with recognized protocols for hemolysis assessment [31,32]. A notable detail that different studies and labs use different hemolysis procedures and readout wavelengths, which could make it challenging to compare results. So, the results should be based on the controls and classification system used in the same experiment [33]. Table 2 shows that Zn had a value of 0.019 (1.06%), which is considered non-toxic by the same standards [32]. Fig. 8 illustrates samples the Toxicity of Zn NPs. We observe from the Table 2 all samples were assessed against the same positive control (1.790). DPPH assay measures how well a test sample can lower the purple DPPH radical by measuring how much light it absorbs (usually at ~517 nm). Formula is used to determine the scavengin: I% = [(A_control − A_sample)/A_control] × 100 [34,35]. Your data consistently uses A_control (DPPH) = 1.137, making it easy to compare inhibitory results between samples and concentrations in the same run. Zn had the most significant result, showing a strong suppression at all concentrations (around 93%–95%) and very low absorbance values (0.061–0.079). This finding indicated almost complete DPPH radical quenching, even at 250 µg/mL. Fig. 9 shows samples the Antioxidant assay of Zn NPs.
This Table 3 presents the descriptive statistical analysis of the toxicity assay for tested compounds at a concentration of 1000 µg/ml. For each compound, the number of replicates was three, and the results are expressed as mean absorbance ± standard deviation, together with the calculated toxicity percentage and the post hoc grouping letters. The mean absorbance values showed a clear variation among the tested compound, indicating different toxicity profiles under the same experimental conditions. The lowest absorbance values were recorded for Zn, with mean values of 0.019 ± 0.003, corresponding to toxicity percentages of 1.04%, respectively. compound were assigned to group c, indicating that the difference them was not statistically significant. These findings suggest that Zn had similarly low toxic effects under the applied conditions, despite Zn showing the numerically lowest toxicity among all tested compounds. The grouping letters provide an important statistical interpretation of these differences, where treatments sharing the same letter are not significantly different, while those with different letters differ significantly. Zn was associated with minimal toxicity.
Table 4 and Table 5 summarizes the antioxidant activity of the tested compounds, namely Zn, at three concentrations: 250, 500, and 1000 µg/ml. For each treatment, the results are presented as the mean absorbance ± standard deviation and the mean inhibition percentage (I%) ± standard deviation, based on three replicates. In addition, significance letters are provided to indicate the statistical grouping among treatments, where values sharing the same letter are not significantly different, whereas values with different letters differ significantly. Overall, the results demonstrate clear differences in antioxidant performance among the tested compounds and across the examined concentrations. Among all treatments, Zn consistently exhibited the highest antioxidant activity, with inhibition percentages of 92.31 ± 0.62%, 94.28 ± 1.14%, and 94.02 ± 0.48% at 250.

CONCLUSION
Zinc nanoparticles were effectively synthesized by a cost-efficient, environmentally benign approach of pulsed laser ablation utilising ginger (Zingiber officinale) extract, a process characterized by Fourier transform analysis. The findings indicated that the synthesized zinc nanoparticles demonstrated antibacterial properties. This phenomenon can be ascribed to the reactive oxygen species generated by the nanoparticles in response to bacteria, along with the electrostatic interactions between the nanoparticles and bacterial species. Moreover, the dimensions of the nanoparticles also affected their antibacterial efficacy. The zinc nanoparticles exhibited greater efficacy against Gram-negative bacterial strains than against Gram-positive pathogens. Consequently, employing an eco-friendly technique to synthesize zinc nanoparticles may substantially influence the treatment of disorders induced by harmful bacteria, thereby presenting prospective uses in the pharmaceutical sector.

CONFLICT OF INTEREST
The authors declare that there is no conflict of interests regarding the publication of this manuscript.