Antibacterial activity of Clinoptilolite/TiO 2 /ZnO photocatalyst against Escherichia coli and Staphylococcus aureus in water resources

Antibacterial activity of Clinoptilolite/TiO 2 /ZnO photocatalyst against Escherichia coli and Staphylococcus aureus in water resources | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Antibacterial activity of Clinoptilolite/TiO 2 /ZnO photocatalyst against Escherichia coli and Staphylococcus aureus in water resources Maryam Abdi, Mansoor Anbia, Maliheh safavi, Mohammad Sepehrian, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2384778/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Water treatment is crucial due to rising water demand across the board, including drinking, agriculture, industry, etc. Pathogens are a type of contamination that must be studied to improve water purification methods. This study investigates the elimination of Escherichia coli ( E. coli ) and Staphylococcus aureus ( S. aureus ) using Clinoptilolite/TiO 2 /ZnO nanocomposite with varied TiO 2 /ZnO ratios of 1:1, 1:2, and 2:1 under UV A and UV C light irradiation. To synergize the effect of coupling TiO 2 with ZnO, we synthesized nanocomposite Clinoptilolite/TiO 2 /ZnO to make a powerful system for water and wastewater disinfection. The characterization tests, including X-ray diffraction, field emission scanning electron microscopy, X-ray energy diffraction, Fourier-transform infrared spectroscopy, diffuse reflection spectroscopy, and nitrogen adsorption-desorption, were performed to confirm the structure. The optimum response for bacterial removal was found by using the determination-optimal (D-optimal) design of response surface methodology (RSM). 100% removal was obtained for E. coli and S. aureus under UV A irradiation and contact time of 100 min for both of them. Biological sciences/Microbiology Earth and environmental sciences/Environmental sciences Health sciences/Health care Antibacterial activity Clinoptilolite/TiO2/ZnO Photocatalyst UV light Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Introduction Water and wastewater treatment have recently received a lot of attention. A highly efficient system for water and wastewater treatment is necessary because of increasing water demands and different usages such as drinking water, agriculture, municipal use and, industry 1 . In the last decade, the pathogens such as bacteria, viruses, fungi, and protozoa have got most of the attention in water sanitation 2 . Waterborne diseases such as fecal contamination cause 3.3% of mortality in the world 3 . The pathogen, like E. coli , are members of the fecal coliform group, E. coli released into the environment from fecal material, which is known as an indicator of fecal contaminant. S. aureus is a species of gram-positive that causes most human diseases. These opportunistic human pathogens, such as S. aureus and E. coli , usually cause watery diarrhea, typhoid, gastroenteritis, neonatal meningitis, urinary tract infection, and Haemolytic-Uraemic Syndrome (HUS) 4 – 6 . There are various water disinfection technologies, such as ozonation, chlorination, UV irradiation, advanced oxidation, membrane techniques, and using photocatalysts 7 . The use of photocatalysts has various benefits, including their widespread availability, low cost, lack of harmful disinfection by-products, and efficacy against microorganisms 8 , 9 . Photocatalysts, as semiconductors that undergo an induced photoreaction, have photocatalytic capabilities that remove microorganisms from water or wastewater. Bacteria can be eliminated using photocatalytic processes using conventional photocatalysts, such as TiO 2 10–13 , ZnO 14 – 16 , CuO 17 , 18 , and Ag 19 – 21 . There are two bands in the mentioned photocatalysts: valence band and the conduction band. The gap between two bands is referred to as the band-gap. Light absorption results in the creation of electrons and holes in the conduction and valence bands. The hole in the valence band can function as an oxidant and create highly reactive hydroxyl radicals. The electrons in the conduction band could undergo successive reduction reactions 22 – 24 . To a large extent, photocatalytic efficiency is dependent on crystal structure. TiO 2 is one of the semiconductors which is extensively applied in water disinfection. TiO 2 has four polymorphs, including monoclinic (TiO 2 B), anatase, brookite, and rutile of which, anatase and rutile are more common. Anatase shows higher stability and photocatalytic activity than rutile. Also, anatase has the longest charge carrier lifetime and causes a higher photocatalytic effect than the other TiO 2 polymorphs 25 . The synthesis method has an effect on the surface area of TiO 2 nanoparticles 26 . The advantages of TiO 2 as a photocatalyst include insolubility in water, non-toxicity, low cost, eco-friendliness, high stability, and accessibility 27 , 28 . ZnO is an inexpensive, highly redox-active, and antibacterial transition metal oxide that is also a well-known photocatalyst 29 . ZnO appears in three crystal structures including wurtzite, zinc blend, and rock salt 30 . A powerful system for water and wastewater disinfection is obtained by synergizing the impact of coupling photocatalysts such as TiO2 with ZnO. ZnO causes charge transfer, so it prevents the recombination of electron holes in TiO 2 31 . Supporting materials such as zeolites, crystalline materials with porous structures, play an essential role in photocatalytic processes 32 , 33 . The two necessary reactants in zeolite synthesis are silicon and aluminum 34 . Using zeolite as support reduces the secondary pollutants of free nanoparticles in water, prevents nanoparticle aggregation, regulates nanoparticle release, and increases contact area 35 . The following is the common mechanism of photocatalytic antibacterial activity: photocatalyst can generate electron-hole pairs under UV irradiation. As a result of the photocatalytic reaction reactive oxygen species (ROS) such as superoxide radicals, hydroxyl, and hydrogen peroxide are produced. Due to the assault of ROS on Coenzyme A in the cell membrane, cellular respiration activity is hindered. ROS has the ability to punch holes in the cell membrane. Because of this, certain vital cations (like K + ) escape, leading to the death of the microorganism 36 , 37 . RSM is an efficient statistical strategy for experimental design. In this study, RSM was applied to optimize the parameters and investigate the independent parameters, including the concentration of photocatalyst (A, mg/mL), contact time (B, min), the ratio of TiO 2 /ZnO (C), and type of UV light (D). This research assessed the efficacy of a new nanocomposite, Clinoptilolite/TiO2/ZnO, for bacterial eradication under UV A and UV B radiation. Material And Methods Materials. All chemical materials were of analytical grade. Ethanol (absolute for analysis, Merck,), Titanium tetrachloride was used as TiO 2 precursor (Merck, purity 98%), Zinc acetate dihydrate was used as ZnO precursor (Merck, purity 98.5%), Sodium hydroxide (Merck,purity 99%), Sodium chloride extra pure (Merck, purity 99.5%), Nitric acid (Merck, purity 65%), Natural Clinoptilolite purchased from Semnan Negin powder company, Iran. Bacterial strains including Escherichia coli PTCC 1330 and Staphylococcus aureus PTCC 1112 were received from the Persian Type Culture Collection (PTCC) in Tehran, Iran, for antimicrobial testing. Mueller Hinton Broth and Brain Heart Infusion (BHI) agar purchased from Condalab, Spain. Clinoptilolite preparation. In order to remove the Clinoptilolite impurities, 10g of Clinoptilolite was rinsed with distilled water, filtered and dried at 110°C for 24h to eliminate impurities. Nitric acid 1M was mixed with zeolite in the ratio of 1:10 and stirred in bath oil at 80°C under reflux reaction for 2h. After that, the mixture was then filtered and washed repeatedly to attain a pH 7. Natural zeolite samples were dried for 24 h at 110°C and then calcined at 500°C for 4h 38 . Synthesis of TiO 2 . Through the use of the sol-gel process, nanoparticles of TiO 2 in the anatase phase were successfully produced. 2mL of TiCl 4 was added dropwise to 20 mL absolute ethanol at 25°C and stirred until a pale-yellow solution was obtained. The solution was stirring continuously for 1h and then, aged for 5 days until gel formation. Following the production of the gel, it was dried for 24h at 80°C and calcined for 1h at 500°C. The white powder is TiO 2 crystals in the anatase phase 39 . Synthesis of nanocomposite Clinoptilolite/TiO 2 /ZnO. The solution mixing method was applied to synthesize Clinoptilolite/TiO 2 /ZnO nanocomposite. The TiO 2 nanoparticles and Zinc acetate dihydrate (with the ratio of 1:1, 2:1, 1:2) immersed in 10 mL ethanol and 10 mL distilled water. Next, 1g Clinoptilolite, and 4cc Nitric acid with pH 3.5 were added to the solution. The mixture was stirred for 5h at 25°C and was dried in an oven at 110°C for 12h. The final powder calcinated in a muffle furnace at 400°C for 4h to obtain nanocomposite Clinoptilolite/TiO 2 /ZnO. The photocatalyst characterization. X-ray diffraction (XRD, Philips1830, Netherland) applied to determine crystal properties of nanocomposite. Scherrer equation (D = 0.89λ/βcosθ) provides the photocatalyst crystallite size 40 . Fourier infrared spectroscopy (FT-IR, Perkinelmer100, USA) performed to determine functional groups. The morphology and size of nanoparticles were analyzed by transmission electron microscopy (TEM, Philips-EM208S- 100 kv, Netherland). Field emission scanning electron (FE-SEM, Tescan Mira3) images taken to analyze the morphology and size of nanoparticles. In addition, energy dispersive X-ray (EDX, Tescan Mira3) images taken to obtain an elemental analysis of photocatalysts. UV-vis diffuse reflectance spectra (DRS, Shimadzu UV1800, Japan) of photocatalysts measured in the wavelength range of 200–800 nm. The specific surface areas and pore size studied by Brunauer-Emmett-Teller at 77 K (BET, Micromeritics ASAP2020). Bacterial culture preparation. A sanitized 50 mL of Muller-Hinton broth medium was inoculated with the E. coli (PTCC 1330) and S. aureus (PTCC 1112) strains, which were then incubated at 37°C with 200 rpm shaking overnight in a shaking incubator until entering log phase. Next, the optical density (OD) of saturated growth media adjusted to 0.5 McFarland (1.5 × 10 8 CFU/mL). Photocatalyst experimental setup. 0.5 mL of bacterial suspension is mentioned in previous part, diluted in tubs containing 4.5 mL of saline water (NaCl 0.9% solution). After serial dilution, the content of the fifth tube in the series was poured into plates. The photocatalysts with different concentrations (2, 6.5, 11, 15.5, and 20 mg/mL) and various ratios of TiO 2 /ZnO (1:1, 2:1, 1:2) added to the plates as a treatment. The plates exposed to UV A (Philips, 40W, 365nm) illumination for 20, 50, and 100min and UV C (Philips, 15W, 254nm) illumination for 5, 10, and 20min. UV light located 30 cm away from plates. After that, 1 mL of treated solution was cultured by spread plate method on the surface of the BHI agar plate and incubated for 13–18 h at 37°C. Finally, to evaluate the antibacterial effect of photocatalyst colony counting was done manually on treated plates. Response surface methodology. The RSM was employed to evaluate the first and higher-order main effects of each of the mentioned factors and to analyze the impact of the factor interactions to optimize further which variables favor maximum bacterial removal. Expert software version 12.0.1.0 (Stat-Ease Inc., Minneapolis, USA) applied to conduct RSM in this research. D-optimal is design type under RSM (Table 1 ). Design-expert is statistical programme that is extremely committed to the execution of experiment design. It provides comparisons, screening, characterization, optimization, and tough parameter designs. Statistics including analysis of variance (ANOVA) and three-dimensional (3D) curves are used to depict the effect of the factors on the response. The D-optimal quadratic model was employed in this investigation to optimize four variables concentration of photocatalyst (A, mg/mL), contact time (B, min), the ratio of TiO 2 /ZnO (C) and type of UV light (D) for maximum bacteria removal. To get optimal levels of the four selected critical factors for maximal bacterial reduction, thirty runs for E. coli removal (Table 2 ) and twenty-six runs for S. aureus removal (Table 3) were performed. Table 1. The experimental parameters and levels. Symbol variables Units Type of variable Levels Level 1 Level 2 Level 3 Level 4 Level 5 A Concentration of photocatalyst mg/mL Numeric 2 6.5 11 15.5 20 B Contact time min Numeric 5 10 20 50 100 C Ratio TiO 2 /ZnO - Numeric 1 * 2 ** 3 *** - − D Type of UV light - Non-Numeric UV A UV C - - − * Clinoptilolite/TiO 2 /ZnO (1:1) ** Clinoptilolite/TiO 2 /ZnO (1:2) *** Clinoptilolite/TiO 2 /ZnO (2:1) Table 2. The actual and predicted value of response for E. coli . Run A: photocatalyst Concentration (mg/mL) B: Contact time (min) C: Ratio TiO 2 /ZnO D: Type of UV light Actual values (CFU/mL) Predicted values (CFU/mL) 1 20 50 1 UV A 0 476.71 2 20 20 1 UV A 200 685.75 3 2 20 1 UV A 500 0 4 2 50 1 UV A 550 852.12 5 11 50 3 UV A 150 4526.46 6 20 20 3 UV A 15500 16015.29 7 2 5 3 UV C 11000 8260.87 8 20 20 3 UV C 1250 2715.69 9 15.5 5 1 UV C 100 1150.12 10 2 5 1 UV C 13200 13347.43 11 20 20 3 UV A 17600 16015.29 12 11 50 2 UV A 3600 366.58 13 15.5 10 3 UV C 150 1484.76 14 20 5 2 UV C 1350 2946.62 15 6.5 100 2 UV A 0 0 16 15.5 5 1 UV C 650 1150.12 17 2 100 3 UV A 1150 1557.02 18 2 5 3 UV C 7450 8260.87 19 6.5 10 3 UV C 2000 3528.84 20 11 100 3 UV A 0 0 21 15.5 10 2 UV C 50 1322.23 22 2 100 1 UV A 0 2288.30 23 20 5 3 UV C 9050 4351.18 24 20 100 2 UV A 2400 3710.49 25 6.5 100 1 UV A 0 0 26 20 20 2 UV C 350 2076.62 27 11 100 3 UV A 2200 0 28 20 20 1 UV C 1600 1437.55 29 20 20 1 UV C 3650 1437.55 30 15.5 20 1 UV C 2800 1179.44 Table 3. The actual and predicted value of response for S. aureus. Run A: photocatalyst Concentration (mg/mL) B: Contact time (min) C: Ratio TiO 2 /ZnO D: Type of UV light Actual values (CFU/mL) Predicted values (CFU/mL) 1 20 50 1 UV A 15800 15795.27 2 20 20 1 UV A 46000 45290.27 3 20 20 3 UV A 95000 105400 4 15.5 100 2 UV A 0 19442.76 5 2 5 3 UV C 110000 116200 6 20 20 3 UV C 47200 57382.40 7 2 20 1 UV C 0 0 8 2 5 1 UV C 124000 129000 9 20 20 3 UV A 115000 105400 10 20 100 1 UV A 34600 34077.04 11 11 50 2 UV A 40000 39750.79 12 15.5 10 3 UV C 37600 37686.72 13 20 5 2 UV C 124000 104200 14 6.5 100 2 UV A 24400 28438.42 15 15.5 5 1 UV C 43200 45245.99 16 2 100 3 UV A 124000 140600 17 2 5 3 UV C 124000 116200 18 6.5 10 3 UV C 28000 22342.29 19 11 100 3 UV A 84000 65441.98 20 15.5 10 2 UV C 21 13212.39 21 2 100 1 UV A 77600 67221.48 22 20 5 3 UV C 124000 132600 23 2 5 2 UV C 115000 111000 24 15.5 5 1 UV C 40600 45245.99 25 20 20 2 UV C 36000 23385.01 26 11 100 3 UV A 76000 65441.98 Results And Discussion XRD characterization. The XRD analysis for nanocomposite Clinoptilolite/TiO 2 /ZnO presented in Fig. 1 2θ is in the range between 5° and 90°. Figure 1 a represents diffraction peaks of raw Clinoptilolite, which are located at 2θ = 10, 11.4, 17, 21, 23, 26, 50, and 68 (JCPDS card No. 25-1349). Figure 1 b represents diffraction peaks of anatase TiO 2 at 2θ = 25.3, 37.8, 47.9, 54 and 55, which are respectively assign to the {101}, {004}, {200}, {204} and {116} crystal plane (JCPDS card No. 78-1349). Figures 1 c, d, and e show nanocomposite Clinoptilolite/TiO 2 /ZnO with different ratios of TiO 2 /ZnO 1:1, 2:1, and 1:2, respectively. Diffraction peaks of ZnO at 2θ = 31.82, 34.47, 36.3, 47.63, 56.65, 62.9, 68.03 and 69.12, which are respectively attributed to the {100}, {002}, {200}, {101}, {102}, {180}, {103}, {200} and {112} crystal plane (JCPDS card No. 36-1451). As shown in Fig. 1 , the intensity of TiO 2 and ZnO in nanocomposite Clinoptilolite/TiO 2 /ZnO with the molar ratio of TiO 2 /ZnO 1:1 is equal. In the 2:1 molar ratio of TiO 2 /ZnO, the peak intensity of TiO 2 is approximately twice that of ZnO, and in the 1:2 molar ratio of TiO 2 /ZnO the peak intensity of ZnO is about twice that of TiO 2 , which is also well seen in the XRD pattern. The XRD pattern demonstrates that the peaks related to support, TiO 2 and ZnO are all observed, which indicates the interaction between support and nanoparticles with no distraction during the synthesis process. Optical properties. To represent the ability of UV absorbance by photocatalyst, UV-vis DRS was performed. As demonstrated in Fig. 2 , photocatalysts which were synthesized in all ratios (TiO 2 /ZnO 1:1, 2:1, and 1:2), revealed optical properties for all wavelengths of UV (UV C = 200–280 nm, UV B = 280–315 nm, and UV A = 315–400 nm). According to the UV-vis DRS result, UV C irradiation was selected as the shortest wavelength, and UV A irradiation selected as the longest wavelength in this research. FT-IR spectra. The FT-IR spectra of nanocomposite Clinoptilolite/TiO 2 /ZnO with different ratios are presented in Fig. 3 . Figure 3 a is related to raw Clinoptilolite. The peak at 839 cm − 1 corresponds to the stretching vibration of Si-O-Al, and a broad peak at 1080 cm − 1 observed, which is related to the stretching vibration of Si-O-Si. The peak at 680 cm − 1 belongs to the stretching vibration of SiO 2 . The weak stretching band near 3500 cm − 1 and the bending vibration band near 1600 cm − 1 correspond to hydroxyl (-OH) groups in Clinoptilolite. Figure 3 b, c, and d are related to nanocomposite Clinoptilolite/TiO 2 /ZnO with different ratios of TiO 2 /ZnO 1:1, 2:1, and 1:2, respectively. The peak appeared at 1066 cm − 1 and corresponded to TiO 2 , which is led to a slight shift toward a shorter wavenumber compared to 1080 cm − 1 , indicating specific chemical interactions between TiO 2 and Clinoptilolite. A slight shift in the hydroxyl group is observed in the nanocomposite corresponding to Clinoptilolite, which is due to the binding of zinc cation in the Clinoptilolite structure during the ion exchange process. The shift in the hydroxyl group occurs because of overlapping between ZnO and hydroxyl groups that are present outside or inside the Clinoptilolite structure. FE-SEM and TEM characterization. Figure 4 illustrates FE-SEM images of the nanocomposite. Figure 4 a illustrates raw Clinoptilolite with a uniform and sheet structure. Figures 4 b, c and d illustrate nanocomposite Clinoptilolite/TiO 2 /ZnO with molar ratios of 1:1, 2:1 and 1:2 respectively. After loading spherical nanoparticles of TiO 2 and ZnO, the surface becomes rough, which indicates nanoparticles were loaded on the Clinoptilolite surface as well. Clinoptilolite prevents the dispersion of nanoparticles in an aqueous medium and prevents secondary contamination. It was observed from TEM image (Fig. 5 a) that the morphology of TiO 2 and ZnO nanoparticles are pseudo-spherical and road-like shape. The average particle size is estimated to be about 36.75 nm (Fig. 5 b). EDX analysis. The EDX spectrum of raw Clinoptilolite was demonstrated in Fig. 6 a. However, Figs. 6 b, c, and d show nanocomposite Clinoptilolite/TiO 2 /ZnO with molar ratios 1:1, 2:1, and 1:2, respectively. At ratio 2:1, the intensity of Ti peaks is twice that of Zn, and at ratio 1:2 the intensity of Zn peaks twice that of Ti approximately. EDX mapping images illustrate the distribution of elements in Fig. 7 . The elements of Ti and Zn were distributed on Clinoptilolite as well. BET measurements. BET analysis provides useful information about porosity and surface area. Figure 8 shows the BET of raw Clinoptilolite, and nanocomposite Clinoptilolite/TiO 2 /ZnO (1:1) at 77 K. According to IUPAC classification, nanocomposite was demonstrated as a typical IV adsorption-desorption isotherm with hysteresis loop which is confirmed the mesoporous structure of nanocomposite. Table 4 summarizes the specific surface area (S BET (m 2 /g)), pore volume (V total (cm 3 /g)), and average pore diameter (d (nm)) by BJH model. As results show that raw Clinoptilolite´s surface area is 99.1101 m 2 /g while the surface area of nanocomposite Clinoptilolite/TiO 2 /ZnO (1:1) is 34.1292 m 2 /g which demonstrated as surface area decrease after compositing. The pore size distribution by the BJH method showed that the pore size increased because of compositing TiO 2 and ZnO with Clinoptilolite. Table 4 Information of N 2 adsorption-desorption analysis. Compound S BET (m 2 /g) V total (cm 3 /g) d (nm) Raw Clinoptilolite 99.1101 0.130309 10.5674 Clinoptilolite/TiO 2 /ZnO (1:1) 34.1292 0.134773 20.935 Antibacterial activity. The antibacterial effect of photocatalyst was tested in contrast to a blank control (treatment without any light irradiation), light control under UV irradiation, and dark control without any light irradiation (both with photocatalyst treatment). The test results showed that no antibacterial activity observed in control conditions. As natural zeolites contain various mineral elements and impurities, the presence of elements with photocatalytic activities examined. To investigate the photocatalytic effect of Clinoptilolite, the antibacterial test performed. With increasing incubation time up to 100 minutes, the results indicated that Clinoptilolite had no antibacterial activity. Analysis of variance (ANOVA) and prediction of the model equation. As recommended by the software, the quadratic order equation was utilized to analyze the responses, and the final equation for the concentration of bacteria was developed as given in “equation (1)” and “equation (2)”. In “equation (1)” and “equation (2)”, R shows the final concentration of bacteria (CFU/mL), A shows concentration of photocatalyst (mg/mL), B shows contact time (min), C shows ratio of TiO 2 /ZnO, and D shows type of UV light. The ANOVA indicates the adequacy and significance of the model. Tables 5 and 6 summarize the quadratic regression model for E. coli and S. aureus , respectively. According to ANOVA for E. coli , the F-value of the model is 11.53, and the P-value is < 0.0001. The F-value of the model for S. aureus is 16.90, and the P-value is 0.0001. Higher F-values and lower P-values indicate a favorable effect of parameters. Among the independent variables, concentration of photocatalyst and contact time had the greatest effect on bacterial removal. For E. coli , the lack of fit is 2.39, which is not significant and the corresponding P-value is > 0.05. The lack of fit for S. aureus , is 3.30 which is also not significant, and the corresponding P-value is > 0.05. The determination coefficient (R 2 ) for E. coli is 0.8585, while for S. aureus it is 0.9482. The results showed that R 2 has a good corresponding with the adjusted R 2 value. The predicted values of final E. coli and S. aureus concentration plotted versus actual values illustrated in Fig. 9 and Fig. 10 , respectively. Table 5. ANOVA for quadratic model ( E. coli ). Source Sum of squares Degree of freedom (df) Mean square F-value P-Value Model 08 + 6.151E 10 07 + E6.151 11.53 < 0.0001 Significant A- Concentration 07 + 2.2386E 1 07 + 2.2386E 4.47 0.0479 B- Time 07 + 2.026E 1 07 + 2.026E 3.80 0.0633 C- Ratio 06 + 4.193E 1 06 + 4.193E 0.7859 0.3864 D- Light 05 + 1.146E 1 05 + 1.146E 0.0215 0.8850 AB 06 + 2.748E 1 06 + 2.748E 0.5151 0.4817 AC 07 + 4/056E 1 07 + 4/056E 7.60 0.0125 AD 07 + 6.133E 1 07 + 6.133E 11.49 0.0031 BC 07 + 2.695E 1 07 + 2.695E 5.05 0.0367 CD 08 + 1.099E 1 08 + 1.099E 20.61 0.0002 A 2 07 + 8.443E 1 07 + 8.443E 15.82 0.0008 Residual 08 + 1.014E 19 06 + 5.336E Lack Of Fit 07 + 8.820E 14 06 + 6.300E 2.39 0.1717 Not significant Pure Error 07 + 1.318E 5 06 + 2.636E Cor Total 08 + 7.164E 29 R 2 = 0.8585, Adj R 2 = 0.7840, Pred R 2 = 0.6551 Table 6. ANOVA for quadratic model ( S. aureus ). Source Sum of squares Degree of freedom (df) Mean square F-value P-Value Model 10 + 4.643E 13 09 + E3.579 16.90 < 0.0001 Significant A- Concentration 08 + 1.651E 1 08 + 1.651E 0.7809 0.3942 B- Time 10 + 1.170E 1 10 + 1.170E 55.34 < 0.0001 C- Ratio 09 + 7.970E 1 09 + 7.970E 37.70 < 0.0001 D- Light 10 + 1.130E 1 10 + 1.130E 53.45 < 0.0001 AB 09 + 2.368E 1 09 + 2.368E 11.20 0.0058 AC 09 + 1.000E 1 09 + 1.000E 4.73 0.0503 AD 09 + 2.776E 1 09 + 2.776E 13.13 0.0035 BC 09 + 1.085E 1 09 + 1.085E 5.13 0.0428 BD 09 + 1.488E 1 09 + 1.488E 7.04 0.0210 CD 07 + 7.409E 1 07 + 7.409E 0.3505 0.5648 A 2 10 + 1.720E 1 10 + 1.720E 81.36 < 0.0001 B 2 08 + 4.995E 1 08 + 4.995E 2.36 0.1502 C 2 08 + 5.181E 1 08 + 5.181E 2.45 0.1434 Residual 09 + 2.537E 12 08 + 2.114E Lack Of Fit 09 + 2.203E 8 08 + 2.754E 3.30 0.01316 Not significant Pure Error 08 + 3.334E 4 07 + 8.335E Cor Total 10 + 4.897E 25 Interactive effects of variables. A 3D response surface graph plotted to elucidate the effect of each independent variable. The graphs demonstrate the interactive impact of concentration and contact time, concentration and ratio and contact time, and ratio to remove E. coli and S. aureus bacteria in water (Figs. 12 , 13 , and 14 ) According to the graph results, we can obtain optimal conditions. Effect of concentration and time interaction. The interactive effect of concentration and time for E. coli was not significant. Figure 11 shows the one-factor plot of E. coli under UV A irradiation at 100 min contact time in a ratio of 1:1 as an optimum condition. According to Fig. 11 , the number of colonies was eliminated by 100% at two concentrations of 2 mg/mL and 6.5 mg/mL. However, 100% removal was predictable at the contact time of 100 min in the ratio of 1:1 over the broad concentration range (Fig. 12 a). Comparing the actual results for E. coli indicate that at a concentration of 20 mg/mL and contact time of 20 min, the number of colonies in ratio 1:1 was 200 CFU/mL (Table. 2, run 2). Increasing contact time from 20 min to 50 min, at a concentration of 20 mg/mL and ratio of 1:1, caused decreasing the number of colonies from 200 CFU/mL to 0 CFU/mL (Table. 2, runs 2 and 1). Also, increasing concentration from 2 mg/mL to 20 mg/mL, at contact time of 50 min and ratio of 1:1, caused decreasing the number of colonies from 550 CFU/mL to 0 CFU/mL (Table. 2, runs 4 and 1). 3D response surface plot of concentration and time for S. aureus under UV A is demonstrated in Fig. 12 b. RSM has predicted that at times above 50 minutes in ratio 1:1 and concentrations between 15.5 mg/mL and 18.5 mg/mL, the number of colonies reaches zero. Also, actual data (Table. 3, run 4) in ratio 1:2 demonstrated that at a concentration of 15.5 mg/mL, and contact time of 100 min, the number of colonies reaches zero. Increasing concentration from 15.5 mg/mL to 18.5 mg/mL and contact time up to 100 min causes a decrease in the number of colonies. The interactive effects of concentration and time for E. coli and S. aureus under UV C were demonstrated in Fig. 12 c and Fig. 12 d, respectively. Interactive effect of concentration and contact time under UV C for E. coli shown that with increasing concentration, the number of colonies decreases but does not reach zero. At a concentration of 15.5 mg/mL and a contact time of 10 min in a ratio of 1:2, the number of colonies reaches 50 CFU/mL (Table 2 , run 21). Interactive effect of concentration and contact time under UV C for S. aureus shown in the ratio of 1:1 (Fig. 12 d). According to RSM prediction, increasing contact time from 10 min to 20 min in the range of 2 mg/mL to 18.5 mg/mL of photocatalyst, the number of colonies reaches zero. Comparing the actual results for S. aureus indicate that at a concentration of 20 mg/mL and contact time of 5 min, the number of colonies in ratio 1:2 was 124000 CFU/mL (Table. 3, run 13). Increasing contact time from 5 min to 10 min, at a concentration of 15.5 mg/mL and ratio of 1:2, caused decreasing the number of colonies from 124000 CFU/mL to 21 CFU/mL (Table. 3, run 20). Also, increasing concentration from 2 mg/mL to 15.5 mg/mL, at contact time of 5 min and ratio of 1:1, caused decreasing the number of colonies from 124000 CFU/mL to 43200 CFU/mL (Table. 3, run 8 and 15). According to the results, an increase in concentration likely produced more ROS, and bacterial cell surfaces had increased contact with ROS, indicating that ROS can disrupt the bacterial membrane. Also, increasing contact time is probably a likely culprit, as the ROS has more opportunity to damage the bacterial membrane with increased contact time. Increasing the concentration of nanoparticles and contact time reduces the number of colonies 41 . Similar research has shown that E. coli requires more contact time than S. aureus to be inactive 13 . Effect of concentration and ratio interaction. Different contact time (20, 50, and 100 min) under UV A examined in this experiment. Figures 13 a and 13 b represented concentration and ratio interaction for E. coli and S. aureus , respectively. It was predicted by RSM that the number of E. coli colonies decreases to zero once the concentration of photocatalyst (ratio 1:1) is between 2 and 19 mg/mL and the contact time exceeds 20 min. The experimental data indicates that at a contact time of 20 min, a concentration of 2 mg/mL, and a ratio of 1:1, the number of colonies was 500 CFU/mL (Table 2 , run 3). Increasing concentration from 2 mg/mL to 20 mg/mL at the same time (20 min) and ratio (1:1) causes a decrease in the number of colonies to 200 CFU/mL (Table 2 , run 2). Moreover, increasing contact time from 20 min to 50 min at the same concentration (20 mg/mL) and ratio (1:1) reduces the number of colonies from 200 CFU/mL to 0 CFU/mL (Table 2 , run 1). The interactive effect of concentration and ratio for S. aureus at a contact time of 100 min is also shown. Colony counts decreased to zero as predicted by RSM over a broad concentration range (6.5–17.5 mg/mL) at a ratio of 1:1. The actual result (Table 3, run 4) revealed that the number of colonies reaches zero at a concentration of 15.5 mg/mL and a ratio of 1:2. Different contact times (5, 10, and 20 min) under UV C are represented in Figs. 13 c and 13 d for E. coli and S.aureus , respectively. For E. coli , at a contact time of 10 min and concentration of 15.5 mg/mL, in a ratio of 1:2, the number of colonies was 50 CFU/mL (Table 2 , run 21). Also, for S.aureus , the minimum number of colonies (21 CFU/mL) observed at a concentration of 15.5 mg/mL and contact time of 10 min, in ratio of 1:2 (Table 3, run 20). According to Fig. 13 d, in ratios of 1:1 and 1:2, at a concentration 11 mg/mL, 100% removal was predicted for S. aureus (contact time 10 min). All of the actual results confirm the plots. Based on results obtained from the graphs, the ratios 1:1 and 1:2 were the most effective at removing E. coli and S. aureus , respectively. It Should be noted that S. aureus is more susceptible to excess zinc oxide nanoparticles in ratio 1:2 42,43 and also E. coli is more susceptible to titanium dioxide nanoparticles in a ratio of 1:1 44 . The photocatalytic activity of TiO 2 -ZnO binary oxide with different molar ratio of TiO 2 and ZnO was examined by Siwi´nska-Stefa´nska et al. The results indicated the sufficient amount of ZnO can suppress recombination, and causes charge transfer carries on very well 45 . Based on similar research, TiO 2 and ZnO as nanocomposite can increase antibacterial activity against E. coli and S. aureus 46 . Previous researches on some pollutant in aqueous solution highlighted that ZnO more efficient than TiO 2 in photocatalytic degradation 47 , 48 . Effect of time and ratio interaction. The interactive effect of time and ratio at different concentrations of photocatalyst (2, 6.5, 11, 15.5, and 20 mg/mL) under UV A for E. coli and S. aureus represented in Figs. 14 a and b, respectively. The optimum conditions for eradicating E. coli (0 CFU/mL), were achieved in a ratio of 1:1, concentration of 2 mg/mL and contact time of 100 min (Table. 2, run 22). The actual result indicates that increasing contact time from 50 min to 100 min at the same concentration (2 mg/mL) and a ratio of (1:1) cause a decrease in the number of colonies from 550 CFU/mL to 0 CFU/mL (Table. 2, runs 4 and 22). Moreover, in ratio 1:1 and concentration of 6.5 mg/mL, at a contact time of 100 min, the number of colonies reaches zero too (Table 2 , run 25). RSM predicted that at a concentration of 11 mg/mL, the number of colonies would reach zero between 64 and 100 mins at a ratio of 1:1. The actual result represented at a concentration of 15.5 mg/mL, contact time of 100 min, and a ratio of 1:2, the number of colonies reaches zero. Interactive effects of time and ratio at different concentrations of photocatalyst (2, 6.5, 11, 15.5, and 20 mg/mL) under UV C were also investigated for E. coli and S. aureus , which are represented in Figs. 14 c and d, respectively. For E. coli , at a concentration of 2 mg/mL and a contact time of 5 min, the number of colonies in two ratios, 1:1 and 2:1, was 13200 CFU/mL and 7450 CFU/mL, respectively (Table 2 , run 10 and run 18). The number of colonies decreased from 7450 CFU/mL to 2000 CFU/mL when the concentration was increased from 2 mg/mL to 6.5 mg/mL and the contact time was increased from 5 to 10 min in a 2:1 ratio (Table 2 , run 18 and run 19). In addition, increasing concentration from 2 mg/mL to 15.5 mg/mL and contact time from 5 to 20 min in a ratio of 1:1 reduced the number of colonies from 13200 CFU/mL to 2800 CFU/mL (Table 2 , run 10 and run 30). According to the results, increasing concentration and contact time cause a decrease in the number of colonies under UV C irradiation. Moreover, increasing contact time from 5 min to 20 min for S. aureus in a ratio of 1:1 caused the number of colonies to reduce from 124000 CFU/mL to 0 CFU/mL (Table. 3, runs 8 and 7) at the same concentration (20 mg/mL). The findings show that contact time length is most likely important in the eradication of diverse bacterial strains because increasing contact time allows more ROS to permeate the bacterial membrane 13 , 36 , 49 . A similar result reported by Azizi-Lalabadi et al. who demonstrated that 2 mg/mL and 3 mg/mL of nanocomposite TiO 2 /ZnO/4A zeolite with a ratio of 2.5:2.5:95% w/w under visible illumination indicated 100% eradication of E. coli and S. aureus , during 24h contact time. Compositing nanoparticles with zeolite enhanced antibacterial activity by regulating nanoparticle release 35 . In comparison to Azizi-Lalabadi et al., the current investigation reported 100% removal of E. coli and S. aureus during a shorter contact period (100 min) under UV A illumination (photocatalyst concentration = 2 and 15.5 mg/mL and ratios of 1:1 and 1:2, respectively). Determination of optimum conditions for maximum bacterial removal using RSM. The main objective of the optimization is to identify the optimal values of parameters for eliminating bacteria from the model. In the optimization step, the desired response goal (bacterial removal efficiency) was chosen to minimize the value of S. aureus and E. coli , and the parameters applied current concentration of photocatalyst, contact time, a ratio of TiO 2 /ZnO, and type of UV light (D) were chosen to be within range. The Design-Expert version 12.0.1.0 (Stat-Ease Inc., Minneapolis, USA) software program found 100 options with a high desirability of 1.00 for achieving optimal circumstances for maximal bacteria eradication. The optimum condition has been obtained at 100 min contact time under UV A irradiation. At 2 mg/mL and 15.5 mg/mL of photocatalyst and at a TiO2/ZnO ratio of 1:1 and 1:2, respectively, 100% eradication of E. coli and S. aureus was achieved (Fig. 15 ). Also, 100% removal of S. aureus was predicted at concentration 11 mg/mL from 64 min to 100 min in ratio 1:1. To verify the adequacy of the model and the reliability of the optimization stage, further tests were conducted under the suggested optimal circumstances using the above-referenced ideal circumstances, we were able to remove 100% of the E. coli and S. aureus from each experiment as expected. These outcomes prove the RSM's efficacy as a useful tool for achieving optimum working circumstances. Conclusion Nanocomposite Clinoptilolite/TiO 2 /ZnO with different ratios of TiO 2 /ZnO (1:1, 2:1, and 1:2) was synthesized. The antibacterial effect of the novel photocatalyst on E. coli and S. aureus bacteria evaluated under UV A and UV B irradiation. The RSM has chosen to optimize the variables and the interactive effect of photocatalyst concentration, contact time, and TiO 2 /ZnO ratio were investigated as the most important parameters. According to the results, contact time duration had a key role in disinfection. In the current research, UV A was more efficient than UV C because it provides sufficient contact time, is less expensive, and consumes less energy. The maximal E. coli and S. aureus elimination was achieved under these ideal circumstances: source light = UV A , minimum concentration of photocatalyst = 2 and 15.5 mg/mL, contact time = 100 mins and ratios 1:1 and 1:2, respectively. Also, 100% removal of S. aureus was predicted at a concentration of 11 mg/mL from 64 min to 100 min in a ratio of 1:1. These optimal results exhibit high fidelity to the experimental test. Declarations Data availability All data that support the findings of this study are available within the article. Acknowledgements The authors are thankful to Research Council of Iran University of Science and Technology (Tehran) for financial support. 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(2017) doi:10.1016/j.watres.2017.10.007. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-2384778","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":163895715,"identity":"1f623591-3150-4b23-80c3-71fb822027ff","order_by":0,"name":"Maryam Abdi","email":"","orcid":"","institution":"Iran University of Science and Technology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Maryam","middleName":"","lastName":"Abdi","suffix":""},{"id":163895717,"identity":"052c6ea1-e710-41eb-abc1-83bfd82709fb","order_by":1,"name":"Mansoor 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09:29:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2384778/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2384778/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":31119288,"identity":"548935a6-4d0c-4653-b6a2-d527c958daa5","added_by":"auto","created_at":"2023-01-04 19:06:43","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":51766,"visible":true,"origin":"","legend":"\u003cp\u003eXRD pattern for: \u003cstrong\u003e(a)\u003c/strong\u003e raw Clinoptilolite, \u003cstrong\u003e(b)\u003c/strong\u003e anatase TiO\u003csub\u003e2\u003c/sub\u003e, \u003cstrong\u003e(c)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1), \u003cstrong\u003e(d)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (2:1), \u003cstrong\u003e(e)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:2).\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/ff89c96c15e10f808f3d2271.png"},{"id":31118019,"identity":"5b4aefee-1c3e-4795-81e7-2b59816dde76","added_by":"auto","created_at":"2023-01-04 18:58:43","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":44237,"visible":true,"origin":"","legend":"\u003cp\u003eUV-vis DRS \u003cstrong\u003e(a)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1), \u003cstrong\u003e(b)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (2:1), \u003cstrong\u003e(c)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:2).\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/c3e9d1109e9dcbee089e3d70.png"},{"id":31119606,"identity":"d2af8fe9-06f2-4fd9-953b-ec25fd6ed32c","added_by":"auto","created_at":"2023-01-04 19:14:43","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":64601,"visible":true,"origin":"","legend":"\u003cp\u003eFT-IR spectrum: \u003cstrong\u003e(a)\u003c/strong\u003e raw Clinoptilolite, \u003cstrong\u003e(b)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1), \u003cstrong\u003e(c)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (2:1), \u003cstrong\u003e(d)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:2).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/e16e081088bc51f46578985a.png"},{"id":31119294,"identity":"24edb216-cf50-4f8e-9a82-dd65e9daca12","added_by":"auto","created_at":"2023-01-04 19:06:43","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":469562,"visible":true,"origin":"","legend":"\u003cp\u003eFE-SEM images: \u003cstrong\u003e(a)\u003c/strong\u003e raw Clinoptilolite, \u003cstrong\u003e(b)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1), \u003cstrong\u003e(c)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (2:1), \u003cstrong\u003e(d)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:2).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/b3e4d00ad2e91e76b6ca2c4e.png"},{"id":31119289,"identity":"61ffb277-b333-4482-9220-7b16b8523910","added_by":"auto","created_at":"2023-01-04 19:06:43","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":240227,"visible":true,"origin":"","legend":"\u003cp\u003eTEM image of \u003cstrong\u003e(a)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1), \u003cstrong\u003e(b)\u003c/strong\u003e corresponding particle size distribution histogram.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/eb4cd53b0969372027245eee.png"},{"id":31118021,"identity":"cfe893e7-652c-4a7a-af0d-01bb4ba986e4","added_by":"auto","created_at":"2023-01-04 18:58:43","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":121869,"visible":true,"origin":"","legend":"\u003cp\u003eEDX images: \u003cstrong\u003e(a)\u003c/strong\u003e raw Clinoptilolite, \u003cstrong\u003e(b)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1), \u003cstrong\u003e(c)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (2:1), \u003cstrong\u003e(d)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:2).\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/30a8ed28a2e28a4d96852ae0.png"},{"id":31119953,"identity":"30ce62d0-8f53-4b0d-a1fc-98578fad8080","added_by":"auto","created_at":"2023-01-04 19:22:43","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":607756,"visible":true,"origin":"","legend":"\u003cp\u003eEDX mapping: \u003cstrong\u003e(a-c)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1), \u003cstrong\u003e(d-f)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (2:1), \u003cstrong\u003e(g-i)\u003c/strong\u003e nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:2).\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/f0b447be835739cabc826139.png"},{"id":31119607,"identity":"e0f81239-a9fe-44b6-ba17-aa8aedec1349","added_by":"auto","created_at":"2023-01-04 19:14:43","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":86067,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a)\u003c/strong\u003e N\u003csub\u003e2\u003c/sub\u003e adsorption-desorption isotherm of Clinoptilolite, \u003cstrong\u003e(b)\u003c/strong\u003e N\u003csub\u003e2\u003c/sub\u003e adsorption-desorption isotherm of nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1).\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/93e426ed60863ee067fa03df.png"},{"id":31118024,"identity":"ef33976d-72a0-41a5-9da7-524baae6620f","added_by":"auto","created_at":"2023-01-04 18:58:43","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":50706,"visible":true,"origin":"","legend":"\u003cp\u003ePlot reveals the distribution of predicted final \u003cem\u003eE. coli \u003c/em\u003econcentration (CFU/mL) vs.\u003c/p\u003e\n\u003cp\u003eactual final \u003cem\u003eE. coli \u003c/em\u003econcentration (CFU/mL) after treated with Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1, 2:1, and 1:2).\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/7be9ac3e1b08bece77f3044c.png"},{"id":31118029,"identity":"46016dd6-99f3-421b-b512-d7e1ca3414b7","added_by":"auto","created_at":"2023-01-04 18:58:43","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":50486,"visible":true,"origin":"","legend":"\u003cp\u003ePlot reveals the distribution of predicted final \u003cem\u003eS. aureus \u003c/em\u003econcentration (CFU/mL) vs. actual final \u003cem\u003eS. aureus \u003c/em\u003econcentration (CFU/mL) after treated with Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1, 2:1, and 1:2).\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/845e16a81d43a0cfcc8b0816.png"},{"id":31119291,"identity":"6f7dc984-7509-4f47-8d05-a71d9965bbc0","added_by":"auto","created_at":"2023-01-04 19:06:43","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":81758,"visible":true,"origin":"","legend":"\u003cp\u003eOne-factor plot for various concentration (2 mg/mL to 20 mg/mL) of Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (\u003cem\u003eE. coli\u003c/em\u003e under UV\u003csub\u003eA\u003c/sub\u003e, contact time = 100 min, ratio 1:1).\u003c/p\u003e","description":"","filename":"11.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/415c83d7859b09dd4e4d499b.png"},{"id":31118032,"identity":"6e19b679-5ab7-4ca2-9582-50696dcf6779","added_by":"auto","created_at":"2023-01-04 18:58:43","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":624789,"visible":true,"origin":"","legend":"\u003cp\u003e3D response surface plot demonstrates effect of various concentrations of Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO and time at ratio 1:1, \u003cstrong\u003e(a)\u003c/strong\u003e for \u003cem\u003eE. coli\u003c/em\u003e under UV\u003csub\u003eA,\u003c/sub\u003e \u003cstrong\u003e(b)\u003c/strong\u003e for \u003cem\u003eS. aureus \u003c/em\u003eunder UV\u003csub\u003eA\u003c/sub\u003e, \u003cstrong\u003e(c)\u003c/strong\u003e for \u003cem\u003eE. coli\u003c/em\u003e under UV\u003csub\u003eC \u003c/sub\u003eand \u003cstrong\u003e(d)\u003c/strong\u003e for \u003cem\u003eS. aureus \u003c/em\u003eunder UV\u003csub\u003eC\u003c/sub\u003e.\u003c/p\u003e","description":"","filename":"12.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/14f260d2f5405b61bbee3ecb.png"},{"id":31118031,"identity":"adeb7102-9c51-4369-8bbb-c351f5dfd4d6","added_by":"auto","created_at":"2023-01-04 18:58:43","extension":"png","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":632048,"visible":true,"origin":"","legend":"\u003cp\u003e3D response surface plot demonstrates the effect of various concentrations of Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO and ratio, \u003cstrong\u003e(a)\u003c/strong\u003e For \u003cem\u003eE. coli\u003c/em\u003e under UV\u003csub\u003eA\u003c/sub\u003e, contact time 20 min \u003cstrong\u003e(b)\u003c/strong\u003e For \u003cem\u003eS. aureus \u003c/em\u003eunder UV\u003csub\u003eA\u003c/sub\u003e, contact time 100 min, \u003cstrong\u003e(c)\u003c/strong\u003e For \u003cem\u003eE. coli\u003c/em\u003e under UV\u003csub\u003eC\u003c/sub\u003e, contact time 20 min and \u003cstrong\u003e(d)\u003c/strong\u003e For \u003cem\u003eS. aureus \u003c/em\u003eunder UV\u003csub\u003eC\u003c/sub\u003e, contact time 10 min.\u003c/p\u003e","description":"","filename":"13.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/0e074979df113cb7e7655f56.png"},{"id":31118033,"identity":"121e10ce-0bca-4caf-ba34-9f221ceeebe4","added_by":"auto","created_at":"2023-01-04 18:58:43","extension":"png","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":592658,"visible":true,"origin":"","legend":"\u003cp\u003e3D response surface plot demonstrate effect of time and ratio for Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO, \u003cstrong\u003e(a)\u003c/strong\u003e For \u003cem\u003eE. coli\u003c/em\u003e under UV\u003csub\u003eA\u003c/sub\u003e, concentration 2 mg/mL \u003cstrong\u003e(b)\u003c/strong\u003e For \u003cem\u003eS. aureus \u003c/em\u003eunder UV\u003csub\u003eA\u003c/sub\u003e, concentration 11 mg/mL, \u003cstrong\u003e(c)\u003c/strong\u003e For \u003cem\u003eE. coli\u003c/em\u003e under UV\u003csub\u003eC\u003c/sub\u003e concentration 20 mg/mL and \u003cstrong\u003e(d)\u003c/strong\u003e For \u003cem\u003eS. aureus \u003c/em\u003eunder UV\u003csub\u003eC\u003c/sub\u003e concentration 2 mg/mL.\u003c/p\u003e","description":"","filename":"14.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/a8e13960bead274b6e46d7a2.png"},{"id":31118030,"identity":"99bf6088-0b74-41a7-98f5-6e52e1eaa880","added_by":"auto","created_at":"2023-01-04 18:58:43","extension":"png","order_by":15,"title":"Figure 15","display":"","copyAsset":false,"role":"figure","size":629002,"visible":true,"origin":"","legend":"\u003cp\u003eThe optimum experimental results under UV\u003csub\u003eA\u003c/sub\u003e at 100 min contact time \u003cstrong\u003e(a)\u003c/strong\u003e\u003cem\u003e E. coli\u003c/em\u003e in light control \u003cstrong\u003e(b)\u003c/strong\u003e \u003cem\u003eE. coli\u003c/em\u003e after treatment (2 mg/mL of Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1)) \u003cstrong\u003e(c)\u003c/strong\u003e \u003cem\u003eS. aureus \u003c/em\u003ein light control \u003cstrong\u003e(d)\u003c/strong\u003e \u003cem\u003eS. aureus \u003c/em\u003eafter treatment (15.5 mg/mL of Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:2)).\u003c/p\u003e","description":"","filename":"15.png","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/41c65d6c6383f12555730acf.png"},{"id":32682493,"identity":"0f2daec2-a804-4522-aae3-edd3293a5ed8","added_by":"auto","created_at":"2023-02-09 05:44:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4630154,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2384778/v1/61266e4c-b8e1-43cc-8d7f-2c9608d93fdf.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Antibacterial activity of Clinoptilolite/TiO 2 /ZnO photocatalyst against Escherichia coli and Staphylococcus aureus in water resources","fulltext":[{"header":"Introduction","content":"\u003cp\u003eWater and wastewater treatment have recently received a lot of attention. A highly efficient system for water and wastewater treatment is necessary because of increasing water demands and different usages such as drinking water, agriculture, municipal use and, industry \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. In the last decade, the pathogens such as bacteria, viruses, fungi, and protozoa have got most of the attention in water sanitation \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Waterborne diseases such as fecal contamination cause 3.3% of mortality in the world \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e. The pathogen, like \u003cem\u003eE. coli\u003c/em\u003e, are members of the fecal coliform group, \u003cem\u003eE. coli\u003c/em\u003e released into the environment from fecal material, which is known as an indicator of fecal contaminant. \u003cem\u003eS. aureus\u003c/em\u003e is a species of gram-positive that causes most human diseases. These opportunistic human pathogens, such as \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eE. coli\u003c/em\u003e, usually cause watery diarrhea, typhoid, gastroenteritis, neonatal meningitis, urinary tract infection, and Haemolytic-Uraemic Syndrome (HUS) \u003csup\u003e\u003cspan additionalcitationids=\"CR5\" citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThere are various water disinfection technologies, such as ozonation, chlorination, UV irradiation, advanced oxidation, membrane techniques, and using photocatalysts \u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. The use of photocatalysts has various benefits, including their widespread availability, low cost, lack of harmful disinfection by-products, and efficacy against microorganisms \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. Photocatalysts, as semiconductors that undergo an induced photoreaction, have photocatalytic capabilities that remove microorganisms from water or wastewater. Bacteria can be eliminated using photocatalytic processes using conventional photocatalysts, such as TiO\u003csub\u003e2\u003c/sub\u003e \u003csup\u003e10\u0026ndash;13\u003c/sup\u003e, ZnO \u003csup\u003e\u003cspan additionalcitationids=\"CR15\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e, CuO \u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e,\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e, and Ag \u003csup\u003e\u003cspan additionalcitationids=\"CR20\" citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. There are two bands in the mentioned photocatalysts: valence band and the conduction band. The gap between two bands is referred to as the band-gap. Light absorption results in the creation of electrons and holes in the conduction and valence bands. The hole in the valence band can function as an oxidant and create highly reactive hydroxyl radicals. The electrons in the conduction band could undergo successive reduction reactions \u003csup\u003e\u003cspan additionalcitationids=\"CR23\" citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. To a large extent, photocatalytic efficiency is dependent on crystal structure. TiO\u003csub\u003e2\u003c/sub\u003e is one of the semiconductors which is extensively applied in water disinfection. TiO\u003csub\u003e2\u003c/sub\u003e has four polymorphs, including monoclinic (TiO\u003csub\u003e2\u003c/sub\u003e B), anatase, brookite, and rutile of which, anatase and rutile are more common. Anatase shows higher stability and photocatalytic activity than rutile. Also, anatase has the longest charge carrier lifetime and causes a higher photocatalytic effect than the other TiO\u003csub\u003e2\u003c/sub\u003e polymorphs \u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. The synthesis method has an effect on the surface area of TiO\u003csub\u003e2\u003c/sub\u003e nanoparticles \u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e. The advantages of TiO\u003csub\u003e2\u003c/sub\u003e as a photocatalyst include insolubility in water, non-toxicity, low cost, eco-friendliness, high stability, and accessibility \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e,\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e. ZnO is an inexpensive, highly redox-active, and antibacterial transition metal oxide that is also a well-known photocatalyst \u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. ZnO appears in three crystal structures including wurtzite, zinc blend, and rock salt \u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e. A powerful system for water and wastewater disinfection is obtained by synergizing the impact of coupling photocatalysts such as TiO2 with ZnO. ZnO causes charge transfer, so it prevents the recombination of electron holes in TiO\u003csub\u003e2\u003c/sub\u003e \u003csup\u003e31\u003c/sup\u003e. Supporting materials such as zeolites, crystalline materials with porous structures, play an essential role in photocatalytic processes \u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e,\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e. The two necessary reactants in zeolite synthesis are silicon and aluminum \u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e. Using zeolite as support reduces the secondary pollutants of free nanoparticles in water, prevents nanoparticle aggregation, regulates nanoparticle release, and increases contact area \u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e. The following is the common mechanism of photocatalytic antibacterial activity: photocatalyst can generate electron-hole pairs under UV irradiation. As a result of the photocatalytic reaction reactive oxygen species (ROS) such as superoxide radicals, hydroxyl, and hydrogen peroxide are produced. Due to the assault of ROS on Coenzyme A in the cell membrane, cellular respiration activity is hindered. ROS has the ability to punch holes in the cell membrane. Because of this, certain vital cations (like K\u003csup\u003e+\u003c/sup\u003e) escape, leading to the death of the microorganism \u003csup\u003e\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e,\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eRSM is an efficient statistical strategy for experimental design. In this study, RSM was applied to optimize the parameters and investigate the independent parameters, including the concentration of photocatalyst (A, mg/mL), contact time (B, min), the ratio of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (C), and type of UV light (D). This research assessed the efficacy of a new nanocomposite, Clinoptilolite/TiO2/ZnO, for bacterial eradication under UV\u003csub\u003eA\u003c/sub\u003e and UV\u003csub\u003eB\u003c/sub\u003e radiation.\u003c/p\u003e"},{"header":"Material And Methods","content":"\u003cp\u003e\u003cstrong\u003eMaterials.\u003c/strong\u003e All chemical materials were of analytical grade. Ethanol (absolute for analysis, Merck,), Titanium tetrachloride was used as TiO\u003csub\u003e2\u003c/sub\u003e precursor (Merck, purity 98%), Zinc acetate dihydrate was used as ZnO precursor (Merck, purity 98.5%), Sodium hydroxide (Merck,purity 99%), Sodium chloride extra pure (Merck, purity 99.5%), Nitric acid (Merck, purity 65%), Natural Clinoptilolite purchased from Semnan Negin powder company, Iran. Bacterial strains including \u003cem\u003eEscherichia coli\u003c/em\u003e PTCC 1330 and \u003cem\u003eStaphylococcus aureus\u003c/em\u003e PTCC 1112 were received from the Persian Type Culture Collection (PTCC) in Tehran, Iran, for antimicrobial testing. Mueller Hinton Broth and Brain Heart Infusion (BHI) agar purchased from Condalab, Spain.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinoptilolite preparation.\u003c/strong\u003e In order to remove the Clinoptilolite impurities, 10g of Clinoptilolite was rinsed with distilled water, filtered and dried at 110\u0026deg;C for 24h to eliminate impurities. Nitric acid 1M was mixed with zeolite in the ratio of 1:10 and stirred in bath oil at 80\u0026deg;C under reflux reaction for 2h. After that, the mixture was then filtered and washed repeatedly to attain a pH 7. Natural zeolite samples were dried for 24 h at 110\u0026deg;C and then calcined at 500\u0026deg;C for 4h \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e38\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSynthesis of TiO\u003c/strong\u003e \u003csub\u003e\u0026nbsp;\u003cstrong\u003e2\u003c/strong\u003e\u0026nbsp;\u003c/sub\u003e. Through the use of the sol-gel process, nanoparticles of TiO\u003csub\u003e2\u003c/sub\u003e in the anatase phase were successfully produced. 2mL of TiCl\u003csub\u003e4\u003c/sub\u003e was added dropwise to 20 mL absolute ethanol at 25\u0026deg;C and stirred until a pale-yellow solution was obtained. The solution was stirring continuously for 1h and then, aged for 5 days until gel formation. Following the production of the gel, it was dried for 24h at 80\u0026deg;C and calcined for 1h at 500\u0026deg;C. The white powder is TiO\u003csub\u003e2\u003c/sub\u003e crystals in the anatase phase \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSynthesis of nanocomposite Clinoptilolite/TiO\u003c/strong\u003e \u003csub\u003e\u0026nbsp;\u003cstrong\u003e2\u003c/strong\u003e\u0026nbsp;\u003c/sub\u003e \u003cstrong\u003e/ZnO.\u003c/strong\u003e The solution mixing method was applied to synthesize Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO nanocomposite. The TiO\u003csub\u003e2\u003c/sub\u003e nanoparticles and Zinc acetate dihydrate (with the ratio of 1:1, 2:1, 1:2) immersed in 10 mL ethanol and 10 mL distilled water. Next, 1g Clinoptilolite, and 4cc Nitric acid with pH 3.5 were added to the solution. The mixture was stirred for 5h at 25\u0026deg;C and was dried in an oven at 110\u0026deg;C for 12h. The final powder calcinated in a muffle furnace at 400\u0026deg;C for 4h to obtain nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThe photocatalyst characterization.\u003c/strong\u003e X-ray diffraction (XRD, Philips1830, Netherland) applied to determine crystal properties of nanocomposite. Scherrer equation (D\u0026thinsp;=\u0026thinsp;0.89\u0026lambda;/\u0026beta;cos\u0026theta;) provides the photocatalyst crystallite size \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e. Fourier infrared spectroscopy (FT-IR, Perkinelmer100, USA) performed to determine functional groups. The morphology and size of nanoparticles were analyzed by transmission electron microscopy (TEM, Philips-EM208S- 100 kv, Netherland). Field emission scanning electron (FE-SEM, Tescan Mira3) images taken to analyze the morphology and size of nanoparticles. In addition, energy dispersive X-ray (EDX, Tescan Mira3) images taken to obtain an elemental analysis of photocatalysts. UV-vis diffuse reflectance spectra (DRS, Shimadzu UV1800, Japan) of photocatalysts measured in the wavelength range of 200\u0026ndash;800 nm. The specific surface areas and pore size studied by Brunauer-Emmett-Teller at 77 K (BET, Micromeritics ASAP2020).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBacterial culture preparation.\u003c/strong\u003e A sanitized 50 mL of Muller-Hinton broth medium was inoculated with the \u003cem\u003eE. coli\u003c/em\u003e (PTCC 1330) and \u003cem\u003eS. aureus\u003c/em\u003e (PTCC 1112) strains, which were then incubated at 37\u0026deg;C with 200 rpm shaking overnight in a shaking incubator until entering log phase. Next, the optical density (OD) of saturated growth media adjusted to 0.5 McFarland (1.5 \u0026times; 10\u003csup\u003e8\u003c/sup\u003e CFU/mL).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePhotocatalyst experimental setup.\u003c/strong\u003e 0.5 mL of bacterial suspension is mentioned in previous part, diluted in tubs containing 4.5 mL of saline water (NaCl 0.9% solution). After serial dilution, the content of the fifth tube in the series was poured into plates. The photocatalysts with different concentrations (2, 6.5, 11, 15.5, and 20 mg/mL) and various ratios of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1, 2:1, 1:2) added to the plates as a treatment. The plates exposed to UV\u003csub\u003eA\u003c/sub\u003e (Philips, 40W, 365nm) illumination for 20, 50, and 100min and UV\u003csub\u003eC\u003c/sub\u003e (Philips, 15W, 254nm) illumination for 5, 10, and 20min. UV light located 30 cm away from plates. After that, 1 mL of treated solution was cultured by spread plate method on the surface of the BHI agar plate and incubated for 13\u0026ndash;18 h at 37\u0026deg;C. Finally, to evaluate the antibacterial effect of photocatalyst colony counting was done manually on treated plates.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResponse surface methodology.\u003c/strong\u003e The RSM was employed to evaluate the first and higher-order main effects of each of the mentioned factors and to analyze the impact of the factor interactions to optimize further which variables favor maximum bacterial removal. Expert software version 12.0.1.0 (Stat-Ease Inc., Minneapolis, USA) applied to conduct RSM in this research. D-optimal is design type under RSM (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Design-expert is statistical programme that is extremely committed to the execution of experiment design. It provides comparisons, screening, characterization, optimization, and tough parameter designs. Statistics including analysis of variance (ANOVA) and three-dimensional (3D) curves are used to depict the effect of the factors on the response. The D-optimal quadratic model was employed in this investigation to optimize four variables concentration of photocatalyst (A, mg/mL), contact time (B, min), the ratio of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (C) and type of UV light (D) for maximum bacteria removal. To get optimal levels of the four selected critical factors for maximal bacterial reduction, thirty runs for \u003cem\u003eE. coli\u003c/em\u003e removal (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e) and twenty-six runs for \u003cem\u003eS. aureus\u003c/em\u003e removal (Table 3) were performed.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cspan dir=\"LTR\"\u003eTable 1.\u003c/span\u003e\u003c/strong\u003e\u003cspan dir=\"LTR\"\u003e\u0026nbsp;The experimental parameters and levels.\u003c/span\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" id=\"Taba\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eSymbol\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003evariables\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eUnits\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eType of variable\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"6\"\u003e\n \u003cp\u003eLevels\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLevel 1\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLevel 2\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLevel 3\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLevel 4\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eLevel 5\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eConcentration of photocatalyst\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003emg/mL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNumeric\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eContact time\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003emin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNumeric\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eRatio TiO\u003csub\u003e2\u003c/sub\u003e/ZnO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNumeric\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003csub\u003e\u0026minus;\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eType of UV light\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNon-Numeric\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003csub\u003e\u0026minus;\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"10\"\u003e* Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"10\"\u003e** Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:2)\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"10\"\u003e*** Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (2:1)\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cspan dir=\"LTR\"\u003eTable 2.\u0026nbsp;\u003c/span\u003e\u003c/strong\u003e\u003cspan dir=\"LTR\"\u003eThe actual and predicted value of response for \u003cem\u003eE. coli\u003c/em\u003e.\u003c/span\u003e\u003c/p\u003e\n\u003ctable border=\"1\" id=\"Tab1\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eRun\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eA: photocatalyst Concentration\u003c/p\u003e\n \u003cp\u003e(mg/mL)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eB: Contact time\u003c/p\u003e\n \u003cp\u003e(min)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eC: Ratio TiO\u003csub\u003e2\u003c/sub\u003e/ZnO\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eD: Type of UV light\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eActual values\u003c/p\u003e\n \u003cp\u003e(CFU/mL)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePredicted values\u003c/p\u003e\n \u003cp\u003e(CFU/mL)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e476.71\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e685.75\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e550\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e852.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4526.46\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16015.29\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8260.87\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1250\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2715.69\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1150.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13347.43\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16015.29\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e366.58\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1484.76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1350\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2946.62\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e650\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1150.12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1557.02\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7450\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8260.87\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3528.84\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1322.23\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2288.30\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9050\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4351.18\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2400\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3710.49\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e350\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2076.62\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1437.55\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3650\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1437.55\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2800\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1179.44\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\u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cbr\u003e\u003c/div\u003e\n\u003cdiv align=\"left\" class=\"colspec\"\u003e\u003cstrong\u003eTable 3.\u003c/strong\u003e The actual and predicted value of response for S. aureus.\u003c/div\u003e\n\u003ctable border=\"1\" id=\"Tab2\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eRun\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eA: photocatalyst Concentration\u003c/p\u003e\n \u003cp\u003e(mg/mL)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eB: Contact time\u003c/p\u003e\n \u003cp\u003e(min)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eC: Ratio TiO\u003csub\u003e2\u003c/sub\u003e/ZnO\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eD: Type of UV light\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eActual values\u003c/p\u003e\n \u003cp\u003e(CFU/mL)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePredicted values\u003c/p\u003e\n \u003cp\u003e(CFU/mL)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15800\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15795.27\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e46000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e45290.27\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e95000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e105400\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19442.76\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e110000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e116200\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e47200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e57382.40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e124000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e129000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e115000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e105400\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e34600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e34077.04\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e40000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e39750.79\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e37600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e37686.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e124000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e104200\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e24400\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e28438.42\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e43200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e45245.99\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e124000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e140600\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e124000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e116200\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e28000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22342.29\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e84000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e65441.98\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13212.39\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e77600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e67221.48\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e124000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e132600\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e115000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e111000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e40600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e45245.99\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eC\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e36000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e23385.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUV\u003csub\u003eA\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e76000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e65441.98\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Results And Discussion","content":"\u003cp\u003e\u003cstrong\u003eXRD characterization.\u003c/strong\u003e The XRD analysis for nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO presented in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e 2\u0026theta; is in the range between 5\u0026deg; and 90\u0026deg;. Figure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003ea represents diffraction peaks of raw Clinoptilolite, which are located at 2\u0026theta;\u0026thinsp;=\u0026thinsp;10, 11.4, 17, 21, 23, 26, 50, and 68 (JCPDS card No. 25-1349). Figure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eb represents diffraction peaks of anatase TiO\u003csub\u003e2\u003c/sub\u003e at 2\u0026theta;\u0026thinsp;=\u0026thinsp;25.3, 37.8, 47.9, 54 and 55, which are respectively assign to the {101}, {004}, {200}, {204} and {116} crystal plane (JCPDS card No. 78-1349). Figures \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003ec, d, and e show nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO with different ratios of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO 1:1, 2:1, and 1:2, respectively. Diffraction peaks of ZnO at 2\u0026theta;\u0026thinsp;=\u0026thinsp;31.82, 34.47, 36.3, 47.63, 56.65, 62.9, 68.03 and 69.12, which are respectively attributed to the {100}, {002}, {200}, {101}, {102}, {180}, {103}, {200} and {112} crystal plane (JCPDS card No. 36-1451). As shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e, the intensity of TiO\u003csub\u003e2\u003c/sub\u003e and ZnO in nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO with the molar ratio of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO 1:1 is equal. In the 2:1 molar ratio of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO, the peak intensity of TiO\u003csub\u003e2\u003c/sub\u003e is approximately twice that of ZnO, and in the 1:2 molar ratio of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO the peak intensity of ZnO is about twice that of TiO\u003csub\u003e2\u003c/sub\u003e, which is also well seen in the XRD pattern. The XRD pattern demonstrates that the peaks related to support, TiO\u003csub\u003e2\u003c/sub\u003e and ZnO are all observed, which indicates the interaction between support and nanoparticles with no distraction during the synthesis process.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eOptical properties.\u003c/strong\u003e To represent the ability of UV absorbance by photocatalyst, UV-vis DRS was performed. As demonstrated in Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, photocatalysts which were synthesized in all ratios (TiO\u003csub\u003e2\u003c/sub\u003e/ZnO 1:1, 2:1, and 1:2), revealed optical properties for all wavelengths of UV (UV\u003csub\u003eC\u003c/sub\u003e = 200\u0026ndash;280 nm, UV\u003csub\u003eB\u003c/sub\u003e = 280\u0026ndash;315 nm, and UV\u003csub\u003eA\u003c/sub\u003e = 315\u0026ndash;400 nm). According to the UV-vis DRS result, UV\u003csub\u003eC\u003c/sub\u003e irradiation was selected as the shortest wavelength, and UV\u003csub\u003eA\u003c/sub\u003e irradiation selected as the longest wavelength in this research.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFT-IR spectra.\u003c/strong\u003e The FT-IR spectra of nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO with different ratios are presented in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. Figure \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003ea is related to raw Clinoptilolite. The peak at 839 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e corresponds to the stretching vibration of Si-O-Al, and a broad peak at 1080 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e observed, which is related to the stretching vibration of Si-O-Si. The peak at 680 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e belongs to the stretching vibration of SiO\u003csub\u003e2\u003c/sub\u003e. The weak stretching band near 3500 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and the bending vibration band near 1600 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e correspond to hydroxyl (-OH) groups in Clinoptilolite. Figure \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eb, c, and d are related to nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO with different ratios of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO 1:1, 2:1, and 1:2, respectively. The peak appeared at 1066 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and corresponded to TiO\u003csub\u003e2\u003c/sub\u003e, which is led to a slight shift toward a shorter wavenumber compared to 1080 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, indicating specific chemical interactions between TiO\u003csub\u003e2\u003c/sub\u003e and Clinoptilolite. A slight shift in the hydroxyl group is observed in the nanocomposite corresponding to Clinoptilolite, which is due to the binding of zinc cation in the Clinoptilolite structure during the ion exchange process. The shift in the hydroxyl group occurs because of overlapping between ZnO and hydroxyl groups that are present outside or inside the Clinoptilolite structure.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFE-SEM and TEM characterization.\u003c/strong\u003e Figure \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e illustrates FE-SEM images of the nanocomposite. Figure \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003ea illustrates raw Clinoptilolite with a uniform and sheet structure. Figures \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eb, c and d illustrate nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO with molar ratios of 1:1, 2:1 and 1:2 respectively. After loading spherical nanoparticles of TiO\u003csub\u003e2\u003c/sub\u003e and ZnO, the surface becomes rough, which indicates nanoparticles were loaded on the Clinoptilolite surface as well. Clinoptilolite prevents the dispersion of nanoparticles in an aqueous medium and prevents secondary contamination. It was observed from TEM image (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003ea) that the morphology of TiO\u003csub\u003e2\u003c/sub\u003e and ZnO nanoparticles are pseudo-spherical and road-like shape. The average particle size is estimated to be about 36.75 nm (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eb).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEDX analysis.\u003c/strong\u003e The EDX spectrum of raw Clinoptilolite was demonstrated in Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003ea. However, Figs. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003eb, c, and d show nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO with molar ratios 1:1, 2:1, and 1:2, respectively. At ratio 2:1, the intensity of Ti peaks is twice that of Zn, and at ratio 1:2 the intensity of Zn peaks twice that of Ti approximately. EDX mapping images illustrate the distribution of elements in Fig. \u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e. The elements of Ti and Zn were distributed on Clinoptilolite as well.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBET measurements.\u003c/strong\u003e BET analysis provides useful information about porosity and surface area. Figure \u003cspan class=\"InternalRef\"\u003e8\u003c/span\u003e shows the BET of raw Clinoptilolite, and nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1) at 77 K. According to IUPAC classification, nanocomposite was demonstrated as a typical IV adsorption-desorption isotherm with hysteresis loop which is confirmed the mesoporous structure of nanocomposite. Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e summarizes the specific surface area (S\u003csub\u003eBET\u003c/sub\u003e (m\u003csup\u003e2\u003c/sup\u003e/g)), pore volume (V\u003csub\u003etotal\u003c/sub\u003e (cm\u003csup\u003e3\u003c/sup\u003e/g)), and average pore diameter (d (nm)) by BJH model. As results show that raw Clinoptilolite\u0026acute;s surface area is 99.1101 m\u003csup\u003e2\u003c/sup\u003e/g while the surface area of nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1) is 34.1292 m\u003csup\u003e2\u003c/sup\u003e/g which demonstrated as surface area decrease after compositing. The pore size distribution by the BJH method showed that the pore size increased because of compositing TiO\u003csub\u003e2\u003c/sub\u003eand ZnO with Clinoptilolite.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable border=\"1\" id=\"Tab3\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eInformation of N\u003csub\u003e2\u003c/sub\u003e adsorption-desorption analysis.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCompound\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eS\u003csub\u003eBET\u003c/sub\u003e (m\u003csup\u003e2\u003c/sup\u003e/g)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eV\u003csub\u003etotal\u003c/sub\u003e (cm\u003csup\u003e3\u003c/sup\u003e/g)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ed (nm)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eRaw Clinoptilolite\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e99.1101\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.130309\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.5674\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eClinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e34.1292\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.134773\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.935\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\u003eAntibacterial activity.\u003c/strong\u003e The antibacterial effect of photocatalyst was tested in contrast to a blank control (treatment without any light irradiation), light control under UV irradiation, and dark control without any light irradiation (both with photocatalyst treatment). The test results showed that no antibacterial activity observed in control conditions. As natural zeolites contain various mineral elements and impurities, the presence of elements with photocatalytic activities examined. To investigate the photocatalytic effect of Clinoptilolite, the antibacterial test performed. With increasing incubation time up to 100 minutes, the results indicated that Clinoptilolite had no antibacterial activity.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAnalysis of variance (ANOVA) and prediction of the model equation.\u003c/strong\u003e As recommended by the software, the quadratic order equation was utilized to analyze the responses, and the final equation for the concentration of bacteria was developed as given in \u0026ldquo;equation (1)\u0026rdquo; and \u0026ldquo;equation (2)\u0026rdquo;.\u003c/p\u003e\n\u003cdiv class=\"Section2\" id=\"Sec4\"\u003e\n \u003cp\u003e\u003cimg 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\" width=\"666\" height=\"307\"\u003e\u003c/p\u003e\n \u003cdiv class=\"Section3\" id=\"Sec5\"\u003e\n \u003cp\u003eIn \u0026ldquo;equation (1)\u0026rdquo; and \u0026ldquo;equation (2)\u0026rdquo;, R shows the final concentration of bacteria (CFU/mL), A shows concentration of photocatalyst (mg/mL), B shows contact time (min), C shows ratio of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO, and D shows type of UV light.\u003c/p\u003e\n \u003cp\u003eThe ANOVA indicates the adequacy and significance of the model. Tables \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e and 6 summarize the quadratic regression model for \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e, respectively. According to ANOVA for \u003cem\u003eE. coli\u003c/em\u003e, the F-value of the model is 11.53, and the P-value is \u0026lt;\u0026thinsp;0.0001. The F-value of the model for \u003cem\u003eS. aureus\u003c/em\u003e is 16.90, and the P-value is 0.0001. Higher F-values and lower P-values indicate a favorable effect of parameters. Among the independent variables, concentration of photocatalyst and contact time had the greatest effect on bacterial removal. For \u003cem\u003eE. coli\u003c/em\u003e, the lack of fit is 2.39, which is not significant and the corresponding P-value is \u0026gt;\u0026thinsp;0.05. The lack of fit for \u003cem\u003eS. aureus\u003c/em\u003e, is 3.30 which is also not significant, and the corresponding P-value is \u0026gt;\u0026thinsp;0.05. The determination coefficient (R\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e) for \u003cem\u003eE. coli\u003c/em\u003e is 0.8585, while for \u003cem\u003eS. aureus\u003c/em\u003e it is 0.9482. The results showed that R\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e has a good corresponding with the adjusted R\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e value. The predicted values of final \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e concentration plotted versus actual values illustrated in Fig. \u003cspan class=\"InternalRef\"\u003e9\u003c/span\u003e and Fig. \u003cspan class=\"InternalRef\"\u003e10\u003c/span\u003e, respectively.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 5.\u003c/strong\u003e ANOVA for quadratic model (\u003cem\u003eE. coli\u003c/em\u003e).\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable border=\"1\" id=\"Tabc\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSource\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSum of squares\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDegree of freedom\u003c/p\u003e\n \u003cp\u003e(df)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean square\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eF-value\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-Value\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eModel\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;6.151E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;E6.151\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.53\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA- Concentration\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;2.2386E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;2.2386E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0479\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB- Time\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;2.026E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;2.026E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0633\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC- Ratio\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e06\u0026thinsp;+\u0026thinsp;4.193E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e06\u0026thinsp;+\u0026thinsp;4.193E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7859\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.3864\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eD- Light\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e05\u0026thinsp;+\u0026thinsp;1.146E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e05\u0026thinsp;+\u0026thinsp;1.146E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0215\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8850\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e06\u0026thinsp;+\u0026thinsp;2.748E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e06\u0026thinsp;+\u0026thinsp;2.748E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.5151\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.4817\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;4/056E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;4/056E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0125\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;6.133E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;6.133E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0031\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;2.695E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;2.695E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0367\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;1.099E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;1.099E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e20.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;8.443E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;8.443E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0008\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eResidual\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;1.014E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e06\u0026thinsp;+\u0026thinsp;5.336E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLack Of Fit\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;8.820E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e06\u0026thinsp;+\u0026thinsp;6.300E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1717\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot significant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePure Error\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;1.318E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e06\u0026thinsp;+\u0026thinsp;2.636E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCor Total\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;7.164E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eR\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.8585, Adj R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.7840, Pred R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.6551\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 6.\u003c/strong\u003e ANOVA for quadratic model (\u003cem\u003eS. aureus\u003c/em\u003e).\u003c/p\u003e\n \u003ctable border=\"1\" id=\"Tab4\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSource\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSum of squares\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eDegree of freedom\u003c/p\u003e\n \u003cp\u003e(df)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMean square\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eF-value\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-Value\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eModel\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u0026thinsp;+\u0026thinsp;4.643E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;E3.579\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e16.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSignificant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA- Concentration\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;1.651E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;1.651E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7809\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.3942\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB- Time\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u0026thinsp;+\u0026thinsp;1.170E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u0026thinsp;+\u0026thinsp;1.170E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e55.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC- Ratio\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;7.970E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;7.970E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e37.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eD- Light\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u0026thinsp;+\u0026thinsp;1.130E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u0026thinsp;+\u0026thinsp;1.130E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e53.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;2.368E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;2.368E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0058\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;1.000E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;1.000E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0503\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;2.776E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;2.776E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0035\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;1.085E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;1.085E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0428\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eBD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;1.488E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;1.488E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0210\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;7.409E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;7.409E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.3505\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.5648\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u0026thinsp;+\u0026thinsp;1.720E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u0026thinsp;+\u0026thinsp;1.720E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e81.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;4.995E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;4.995E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1502\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eC\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;5.181E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;5.181E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1434\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eResidual\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;2.537E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;2.114E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLack Of Fit\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e09\u0026thinsp;+\u0026thinsp;2.203E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;2.754E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.01316\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot significant\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePure Error\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e08\u0026thinsp;+\u0026thinsp;3.334E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e07\u0026thinsp;+\u0026thinsp;8.335E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCor Total\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u0026thinsp;+\u0026thinsp;4.897E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003cstrong\u003eInteractive effects of variables.\u003c/strong\u003e A 3D response surface graph plotted to elucidate the effect of each independent variable. The graphs demonstrate the interactive impact of concentration and contact time, concentration and ratio and contact time, and ratio to remove \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e bacteria in water (Figs. \u003cspan class=\"InternalRef\"\u003e12\u003c/span\u003e, \u003cspan class=\"InternalRef\"\u003e13\u003c/span\u003e, and \u003cspan class=\"InternalRef\"\u003e14\u003c/span\u003e) According to the graph results, we can obtain optimal conditions.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eEffect of concentration and time interaction.\u003c/strong\u003e The interactive effect of concentration and time for \u003cem\u003eE. coli\u003c/em\u003e was not significant. Figure \u003cspan class=\"InternalRef\"\u003e11\u003c/span\u003e shows the one-factor plot of \u003cem\u003eE. coli\u003c/em\u003e under UV\u003csub\u003eA\u003c/sub\u003e irradiation at 100 min contact time in a ratio of 1:1 as an optimum condition. According to Fig. \u003cspan class=\"InternalRef\"\u003e11\u003c/span\u003e, the number of colonies was eliminated by 100% at two concentrations of 2 mg/mL and 6.5 mg/mL. However, 100% removal was predictable at the contact time of 100 min in the ratio of 1:1 over the broad concentration range (Fig. \u003cspan class=\"InternalRef\"\u003e12\u003c/span\u003ea). Comparing the actual results for \u003cem\u003eE. coli\u003c/em\u003e indicate that at a concentration of 20 mg/mL and contact time of 20 min, the number of colonies in ratio 1:1 was 200 CFU/mL (Table. 2, run 2). Increasing contact time from 20 min to 50 min, at a concentration of 20 mg/mL and ratio of 1:1, caused decreasing the number of colonies from 200 CFU/mL to 0 CFU/mL (Table. 2, runs 2 and 1). Also, increasing concentration from 2 mg/mL to 20 mg/mL, at contact time of 50 min and ratio of 1:1, caused decreasing the number of colonies from 550 CFU/mL to 0 CFU/mL (Table. 2, runs 4 and 1). 3D response surface plot of concentration and time for \u003cem\u003eS. aureus\u003c/em\u003e under UV\u003csub\u003eA\u003c/sub\u003e is demonstrated in Fig. \u003cspan class=\"InternalRef\"\u003e12\u003c/span\u003eb. RSM has predicted that at times above 50 minutes in ratio 1:1 and concentrations between 15.5 mg/mL and 18.5 mg/mL, the number of colonies reaches zero. Also, actual data (Table. 3, run 4) in ratio 1:2 demonstrated that at a concentration of 15.5 mg/mL, and contact time of 100 min, the number of colonies reaches zero. Increasing concentration from 15.5 mg/mL to 18.5 mg/mL and contact time up to 100 min causes a decrease in the number of colonies. The interactive effects of concentration and time for \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e under UV\u003csub\u003eC\u003c/sub\u003e were demonstrated in Fig. \u003cspan class=\"InternalRef\"\u003e12\u003c/span\u003ec and Fig. \u003cspan class=\"InternalRef\"\u003e12\u003c/span\u003ed, respectively. Interactive effect of concentration and contact time under UV\u003csub\u003eC\u003c/sub\u003e for \u003cem\u003eE. coli\u003c/em\u003e shown that with increasing concentration, the number of colonies decreases but does not reach zero. At a concentration of 15.5 mg/mL and a contact time of 10 min in a ratio of 1:2, the number of colonies reaches 50 CFU/mL (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, run 21). Interactive effect of concentration and contact time under UV\u003csub\u003eC\u003c/sub\u003e for \u003cem\u003eS. aureus\u003c/em\u003e shown in the ratio of 1:1 (Fig. \u003cspan class=\"InternalRef\"\u003e12\u003c/span\u003ed). According to RSM prediction, increasing contact time from 10 min to 20 min in the range of 2 mg/mL to 18.5 mg/mL of photocatalyst, the number of colonies reaches zero. Comparing the actual results for \u003cem\u003eS. aureus\u003c/em\u003e indicate that at a concentration of 20 mg/mL and contact time of 5 min, the number of colonies in ratio 1:2 was 124000 CFU/mL (Table. 3, run 13). Increasing contact time from 5 min to 10 min, at a concentration of 15.5 mg/mL and ratio of 1:2, caused decreasing the number of colonies from 124000 CFU/mL to 21 CFU/mL (Table. 3, run 20). Also, increasing concentration from 2 mg/mL to 15.5 mg/mL, at contact time of 5 min and ratio of 1:1, caused decreasing the number of colonies from 124000 CFU/mL to 43200 CFU/mL (Table. 3, run 8 and 15). According to the results, an increase in concentration likely produced more ROS, and bacterial cell surfaces had increased contact with ROS, indicating that ROS can disrupt the bacterial membrane. Also, increasing contact time is probably a likely culprit, as the ROS has more opportunity to damage the bacterial membrane with increased contact time. Increasing the concentration of nanoparticles and contact time reduces the number of colonies \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e41\u003c/span\u003e\u003c/sup\u003e. Similar research has shown that \u003cem\u003eE. coli\u003c/em\u003e requires more contact time than \u003cem\u003eS. aureus\u003c/em\u003e to be inactive \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eEffect of concentration and ratio interaction.\u003c/strong\u003e Different contact time (20, 50, and 100 min) under UV\u003csub\u003eA\u003c/sub\u003e examined in this experiment. Figures \u003cspan class=\"InternalRef\"\u003e13\u003c/span\u003ea and \u003cspan class=\"InternalRef\"\u003e13\u003c/span\u003eb represented concentration and ratio interaction for \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e, respectively. It was predicted by RSM that the number of \u003cem\u003eE. coli\u003c/em\u003e colonies decreases to zero once the concentration of photocatalyst (ratio 1:1) is between 2 and 19 mg/mL and the contact time exceeds 20 min. The experimental data indicates that at a contact time of 20 min, a concentration of 2 mg/mL, and a ratio of 1:1, the number of colonies was 500 CFU/mL (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, run 3). Increasing concentration from 2 mg/mL to 20 mg/mL at the same time (20 min) and ratio (1:1) causes a decrease in the number of colonies to 200 CFU/mL (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, run 2). Moreover, increasing contact time from 20 min to 50 min at the same concentration (20 mg/mL) and ratio (1:1) reduces the number of colonies from 200 CFU/mL to 0 CFU/mL (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, run 1). The interactive effect of concentration and ratio for \u003cem\u003eS. aureus\u003c/em\u003e at a contact time of 100 min is also shown. Colony counts decreased to zero as predicted by RSM over a broad concentration range (6.5\u0026ndash;17.5 mg/mL) at a ratio of 1:1. The actual result (Table 3, run 4) revealed that the number of colonies reaches zero at a concentration of 15.5 mg/mL and a ratio of 1:2. Different contact times (5, 10, and 20 min) under UV\u003csub\u003eC\u003c/sub\u003e are represented in Figs. \u003cspan class=\"InternalRef\"\u003e13\u003c/span\u003ec and \u003cspan class=\"InternalRef\"\u003e13\u003c/span\u003ed for \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS.aureus\u003c/em\u003e, respectively. For \u003cem\u003eE. coli\u003c/em\u003e, at a contact time of 10 min and concentration of 15.5 mg/mL, in a ratio of 1:2, the number of colonies was 50 CFU/mL (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, run 21). Also, for \u003cem\u003eS.aureus\u003c/em\u003e, the minimum number of colonies (21 CFU/mL) observed at a concentration of 15.5 mg/mL and contact time of 10 min, in ratio of 1:2 (Table 3, run 20). According to Fig. \u003cspan class=\"InternalRef\"\u003e13\u003c/span\u003ed, in ratios of 1:1 and 1:2, at a concentration 11 mg/mL, 100% removal was predicted for \u003cem\u003eS. aureus\u003c/em\u003e (contact time 10 min). All of the actual results confirm the plots. Based on results obtained from the graphs, the ratios 1:1 and 1:2 were the most effective at removing \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e, respectively. It Should be noted that \u003cem\u003eS. aureus\u003c/em\u003e is more susceptible to excess zinc oxide nanoparticles in ratio 1:2 \u003csup\u003e42,43\u003c/sup\u003e and also \u003cem\u003eE. coli\u003c/em\u003e is more susceptible to titanium dioxide nanoparticles in a ratio of 1:1 \u003csup\u003e44\u003c/sup\u003e. The photocatalytic activity of TiO\u003csub\u003e2\u003c/sub\u003e-ZnO binary oxide with different molar ratio of TiO\u003csub\u003e2\u003c/sub\u003e and ZnO was examined by Siwi\u0026acute;nska-Stefa\u0026acute;nska et al. The results indicated the sufficient amount of ZnO can suppress recombination, and causes charge transfer carries on very well \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/sup\u003e. Based on similar research, TiO\u003csub\u003e2\u003c/sub\u003e and ZnO as nanocomposite can increase antibacterial activity against \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e. Previous researches on some pollutant in aqueous solution highlighted that ZnO more efficient than TiO\u003csub\u003e2\u003c/sub\u003e in photocatalytic degradation \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e47\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e48\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eEffect of time and ratio interaction.\u003c/strong\u003e The interactive effect of time and ratio at different concentrations of photocatalyst (2, 6.5, 11, 15.5, and 20 mg/mL) under UV\u003csub\u003eA\u003c/sub\u003e for \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e represented in Figs. \u003cspan class=\"InternalRef\"\u003e14\u003c/span\u003ea and b, respectively. The optimum conditions for eradicating \u003cem\u003eE. coli\u003c/em\u003e (0 CFU/mL), were achieved in a ratio of 1:1, concentration of 2 mg/mL and contact time of 100 min (Table. 2, run 22). The actual result indicates that increasing contact time from 50 min to 100 min at the same concentration (2 mg/mL) and a ratio of (1:1) cause a decrease in the number of colonies from 550 CFU/mL to 0 CFU/mL (Table. 2, runs 4 and 22). Moreover, in ratio 1:1 and concentration of 6.5 mg/mL, at a contact time of 100 min, the number of colonies reaches zero too (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, run 25). RSM predicted that at a concentration of 11 mg/mL, the number of colonies would reach zero between 64 and 100 mins at a ratio of 1:1. The actual result represented at a concentration of 15.5 mg/mL, contact time of 100 min, and a ratio of 1:2, the number of colonies reaches zero. Interactive effects of time and ratio at different concentrations of photocatalyst (2, 6.5, 11, 15.5, and 20 mg/mL) under UV\u003csub\u003eC\u003c/sub\u003e were also investigated for \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e, which are represented in Figs. \u003cspan class=\"InternalRef\"\u003e14\u003c/span\u003ec and d, respectively. For \u003cem\u003eE. coli\u003c/em\u003e, at a concentration of 2 mg/mL and a contact time of 5 min, the number of colonies in two ratios, 1:1 and 2:1, was 13200 CFU/mL and 7450 CFU/mL, respectively (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, run 10 and run 18). The number of colonies decreased from 7450 CFU/mL to 2000 CFU/mL when the concentration was increased from 2 mg/mL to 6.5 mg/mL and the contact time was increased from 5 to 10 min in a 2:1 ratio (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, run 18 and run 19). In addition, increasing concentration from 2 mg/mL to 15.5 mg/mL and contact time from 5 to 20 min in a ratio of 1:1 reduced the number of colonies from 13200 CFU/mL to 2800 CFU/mL (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, run 10 and run 30). According to the results, increasing concentration and contact time cause a decrease in the number of colonies under UV\u003csub\u003eC\u003c/sub\u003e irradiation. Moreover, increasing contact time from 5 min to 20 min for \u003cem\u003eS. aureus\u003c/em\u003e in a ratio of 1:1 caused the number of colonies to reduce from 124000 CFU/mL to 0 CFU/mL (Table. 3, runs 8 and 7) at the same concentration (20 mg/mL). The findings show that contact time length is most likely important in the eradication of diverse bacterial strains because increasing contact time allows more ROS to permeate the bacterial membrane \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e13\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e36\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e49\u003c/span\u003e\u003c/sup\u003e. A similar result reported by Azizi-Lalabadi et al. who demonstrated that 2 mg/mL and 3 mg/mL of nanocomposite TiO\u003csub\u003e2\u003c/sub\u003e/ZnO/4A zeolite with a ratio of 2.5:2.5:95% w/w under visible illumination indicated 100% eradication of \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e, during 24h contact time. Compositing nanoparticles with zeolite enhanced antibacterial activity by regulating nanoparticle release \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e. In comparison to Azizi-Lalabadi et al., the current investigation reported 100% removal of \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e during a shorter contact period (100 min) under UV\u003csub\u003eA\u003c/sub\u003e illumination (photocatalyst concentration\u0026thinsp;=\u0026thinsp;2 and 15.5 mg/mL and ratios of 1:1 and 1:2, respectively).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eDetermination of optimum conditions for maximum bacterial removal using RSM.\u003c/strong\u003e The main objective of the optimization is to identify the optimal values of parameters for eliminating bacteria from the model. In the optimization step, the desired response goal (bacterial removal efficiency) was chosen to minimize the value of \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eE. coli\u003c/em\u003e, and the parameters applied current concentration of photocatalyst, contact time, a ratio of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO, and type of UV light (D) were chosen to be within range. The Design-Expert version 12.0.1.0 (Stat-Ease Inc., Minneapolis, USA) software program found 100 options with a high desirability of 1.00 for achieving optimal circumstances for maximal bacteria eradication. The optimum condition has been obtained at 100 min contact time under UV\u003csub\u003eA\u003c/sub\u003e irradiation. At 2 mg/mL and 15.5 mg/mL of photocatalyst and at a TiO2/ZnO ratio of 1:1 and 1:2, respectively, 100% eradication of \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e was achieved (Fig. \u003cspan class=\"InternalRef\"\u003e15\u003c/span\u003e). Also, 100% removal of \u003cem\u003eS. aureus\u003c/em\u003e was predicted at concentration 11 mg/mL from 64 min to 100 min in ratio 1:1. To verify the adequacy of the model and the reliability of the optimization stage, further tests were conducted under the suggested optimal circumstances using the above-referenced ideal circumstances, we were able to remove 100% of the \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e from each experiment as expected. These outcomes prove the RSM\u0026apos;s efficacy as a useful tool for achieving optimum working circumstances.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eNanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO with different ratios of TiO\u003csub\u003e2\u003c/sub\u003e/ZnO (1:1, 2:1, and 1:2) was synthesized. The antibacterial effect of the novel photocatalyst on \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e bacteria evaluated under UV\u003csub\u003eA\u003c/sub\u003e and UV\u003csub\u003eB\u003c/sub\u003e irradiation. The RSM has chosen to optimize the variables and the interactive effect of photocatalyst concentration, contact time, and TiO\u003csub\u003e2\u003c/sub\u003e/ZnO ratio were investigated as the most important parameters. According to the results, contact time duration had a key role in disinfection. In the current research, UV\u003csub\u003eA\u003c/sub\u003e was more efficient than UV\u003csub\u003eC\u003c/sub\u003e because it provides sufficient contact time, is less expensive, and consumes less energy. The maximal \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e elimination was achieved under these ideal circumstances: source light\u0026thinsp;=\u0026thinsp;UV\u003csub\u003eA\u003c/sub\u003e, minimum concentration of photocatalyst\u0026thinsp;=\u0026thinsp;2 and 15.5 mg/mL, contact time\u0026thinsp;=\u0026thinsp;100 mins and ratios 1:1 and 1:2, respectively. Also, 100% removal of \u003cem\u003eS. aureus\u003c/em\u003e was predicted at a concentration of 11 mg/mL from 64 min to 100 min in a ratio of 1:1. These optimal results exhibit high fidelity to the experimental test.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data that support the findings of this study are available within the article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are thankful to Research Council of Iran University of Science and Technology (Tehran) for financial support.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMaryam Abdi: writing of manuscript, data collection, analyzed the data, designing, carried out experiments. Mansoor Anbia: supervised the study and project administration. Maliheh Safavi: advising antibacterial experiments, review and editing. Mohammad Sepehrian: provided a guidance of the theory, guiding some of experiments. Elham Nezafatian: performed the statistical analysis.\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"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003ePiadeh, F., Moghaddam, M. R. A. \u0026amp; Mardan, S. Present situation of wastewater treatment in the Iranian industrial estates: Recycle and reuse as a solution for achieving goals of eco-industrial parks. \u003cem\u003eResour. Conserv. Recycl.\u003c/em\u003e \u003cstrong\u003e92\u003c/strong\u003e, 172\u0026ndash;178 (2014).\u003c/li\u003e\n\u003cli\u003eJacangelo, J. G., Trussell, R. R., Harza, M. W. \u0026amp; Lane, S. M. Presented, Followed By a Discussion of. 1\u0026ndash;15 (1999).\u003c/li\u003e\n\u003cli\u003eWorld Health Organization (WHO). Water, Sanitation, Hygiene and Health: A Primer for Health Professionals. \u003cem\u003eWorld Heal. 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(2017) doi:10.1016/j.watres.2017.10.007. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Antibacterial activity, Clinoptilolite/TiO2/ZnO, Photocatalyst, UV light","lastPublishedDoi":"10.21203/rs.3.rs-2384778/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2384778/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eWater treatment is crucial due to rising water demand across the board, including drinking, agriculture, industry, etc. Pathogens are a type of contamination that must be studied to improve water purification methods. This study investigates the elimination of \u003cem\u003eEscherichia coli\u003c/em\u003e (\u003cem\u003eE. coli\u003c/em\u003e) and \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (\u003cem\u003eS. aureus\u003c/em\u003e) using Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO nanocomposite with varied TiO\u003csub\u003e2\u003c/sub\u003e/ZnO ratios of 1:1, 1:2, and 2:1 under UV\u003csub\u003eA\u003c/sub\u003e and UV\u003csub\u003eC\u003c/sub\u003e light irradiation. To synergize the effect of coupling TiO\u003csub\u003e2\u003c/sub\u003e with ZnO, we synthesized nanocomposite Clinoptilolite/TiO\u003csub\u003e2\u003c/sub\u003e/ZnO to make a powerful system for water and wastewater disinfection. The characterization tests, including X-ray diffraction, field emission scanning electron microscopy, X-ray energy diffraction, Fourier-transform infrared spectroscopy, diffuse reflection spectroscopy, and nitrogen adsorption-desorption, were performed to confirm the structure. The optimum response for bacterial removal was found by using the determination-optimal (D-optimal) design of response surface methodology (RSM). 100% removal was obtained for \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eS. aureus\u003c/em\u003e under UV\u003csub\u003eA\u003c/sub\u003e irradiation and contact time of 100 min for both of them.\u003c/p\u003e","manuscriptTitle":"Antibacterial activity of Clinoptilolite/TiO 2 /ZnO photocatalyst against Escherichia coli and Staphylococcus aureus in water resources","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-01-04 18:58:37","doi":"10.21203/rs.3.rs-2384778/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9b5eb5c9-66ab-4864-8dce-f56f0eebb78c","owner":[],"postedDate":"January 4th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":18045134,"name":"Biological sciences/Microbiology"},{"id":18045135,"name":"Earth and environmental sciences/Environmental sciences"},{"id":18045136,"name":"Health sciences/Health care"}],"tags":[],"updatedAt":"2023-02-09T05:44:32+00:00","versionOfRecord":[],"versionCreatedAt":"2023-01-04 18:58:37","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2384778","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2384778","identity":"rs-2384778","version":["v1"]},"buildId":"WrCJVZZCHTDjtuVLN7oU0","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}