Photo-oxidative degradation of disposable mask and the effect on the biodegradation process in soil system

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In the terrestrial environment, disposable masks will experience various stages of the degradation process which are influenced by abiotic factors involving sunlight and air as well as biotic factors involving organisms in the soil. There are not many studies that reveal the success of the degradation of disposable masks in nature, so this research was conducted to look at the influence of natural factors in the degradation process of disposable masks. The experiment, which was carried out for 100 days, involved pseudomonas bacteria , worms and plants to condition the degradation media to resemble natural conditions. Changes in disposable masks due to chemical and biological degradation are shown from the results of use tests using microscope, thickness, FTIR and SEM. The results of the analysis showed that there was significant damage to disposable masks through examination with a microscope during the treatment of the masks using a photo oxidation process followed by treatment involving worms and bacteria. Conversely, the formation of important ketone, ester and hydroxide groups after the photooxidation process as shown by the FTIR results have not significant gap with the result in mask without photo-oxidation treatment. Furthermore, Pseudomonas bacteria were able to reduce the thickness of disposable masks effectively compared to single treatment using worms. This research shows the very important role of oxygen availability and sunlight in the degradation process of disposable masks before they reach the complete stage of degradation. biodegradation disposable mask photo-oxidative degradation soil system Figures Figure 1 Figure 2 Figure 3 Introduction The surge in disposable mask waste can be attributed to the heightened demand resulting from the COVID-19 pandemic. The production of masks has escalated in response to the growing demand, which is closely correlated with daily usage. Face masks are utilized to safeguard Public Health against the transmission of diseases. The global monthly usage of masks has surged to 129 billion, with projections indicating a further upward trend (Wang et al., 2020 ). On a daily basis, individuals dispose of around 15.5 trillion tons of mask garbage, which corresponds to 2 million tons of plastic waste. Each mask weighs around 15.5 grams (Wang et al., 2020 ). Hence, the surge in discarded face masks has emerged as a worldwide contributor to plastic contamination. The daily quantity of disposable mask garbage in Indonesia ranges from 53,840 to 80,759 million pieces. Approximately 10 million masks are projected to be discarded on a monthly basis, with an expected total of 1.56 billion masks being disposed of in the year 2020. Disposable masks mostly comprise polypropylene, polyacrylonitrile, and polystyrene polymers (Akber Abbasi et al., 2020 ). Multiple studies have proven that mask waste is a contributing factor to plastic pollution. The COVID-19 epidemic has worsened the problem of plastic pollution, with thousands of discarded masks being disposed of in the environment. This has resulted in potential toxicological risks, particularly for marine environments (Luna et al., 2015 ). Mask manufacture involves the use of chemicals that can undergo decomposition or degradation as a result of physical, chemical, or biological interactions with the natural environment. This particular variant of a three-layer disposable mask offers antibacterial protection, effectively mitigating the transmission of diseases through droplets emitted by individuals in close proximity. This mask serves multiple functions, including filtration, moisture absorption, and water resistance. According to (Farzaneh and Shirinbayan, 2022 ), its effectiveness has been demonstrated and is claimed to be comparable to that of a N95 respirator. However, it is unable to block extremely minute droplets. A disposable mask is composed of three layers: an inner layer that is hydroscopic, a middle layer that acts as a fine filter, and a back layer that is water resistant. The mask consists of three layers: the outer layer is composed of spun-bonded non-woven fabric, the middle layer is made of melt-blown non-woven fabric, and the inner layer is also spun-bonded non-woven fabric (J. Wang et al., 2023 ). Non-woven materials, like polypropylene, possess notable attributes such as high temperature tolerance and a structurally stable composition. In addition, non-woven materials are single-use only. The intermolecular connections between the fibers of the mask are strengthened as a result of the introduction of exogenous substances. Disposable masks can undergo biodegradation in the soil, facilitated by the actions of living organisms (Lucas et al., 2008 ). Nevertheless, disposable masks are typically composed of polypropylene or polyethylene, both of which are robust polymers that exhibit limited degradability. This can result in an extended duration for the natural degradation process, potentially exceeding a decade (Canopoli et al., 2020 ). Consequently, multiple techniques have been devised to accelerate this process of degradation. Liu et al., ( 2022 ) conducted an experiment to assess the properties of disposable masks following exposure to UV radiation. The results revealed a reduction in elongation at break, tensile strength, and maximum force of the masks, suggesting the occurrence of chemical bond breakdown. A recent study conducted by Y. Wang et al., ( 2023 ) employed the oxo degradation technique, utilizing O 3 and H 2 O 2 , to facilitate the degradation of polyethylene. The investigation revealed alterations in the polymer bonds, rendering them more susceptible to breakage. The combination of UV radiation with oxidation chemicals, such as hydrogen peroxide, can generate hydroxyl radicals. These radicals facilitate the degradation of the polymer bonds. Easton et al., ( 2023 ) documented the development of superficial cavities, depressions, and cracks on the surface of the microplastic, as well as alterations in the proportion of oxygen-containing functional groups, following its exposure to UV/ H 2 O 2 treatment. The presence of cavities, cracks, and alterations in polymer linkages that arise during the photo-oxo degradation process will facilitate the acceleration of the biodegradation process. The objective of this study is to assess the impact of photo-oxo degradation on the biodegradation process inside the soil system. In order to replicate a natural biodegradation environment, single-use masks were placed in a pot along with pseudomona s bacteria, earthworms, and plants for a duration of over 100 days. The masks underwent evaluation of their physical and chemical alterations following treatment by the use of SEM, Microscope, micrometer screw gauge, and FTIR analysis. By integrating the photo-oxo degradation and biodegradation mechanisms, we aim to address the escalating issue of mask waste, which has been exacerbated by the COVID-19 pandemic. Materials and Method 2.1 Materials The sample used is the Sensi mask of a surgical face mask, a 3-ply earloop, SENSI brand, obtained from Arista Latino Company, Indonesia, standardized for Disposable Mask Characteristics (SNI EN 14683:2019; AC: 2019; and SNI 8488:2018). The soil medium used is loamy soil. The compost used was Trubus compost obtained from a plant store. Additional worms used were Lumbricus rubellus and Capsicum annuum L plants obtained from plant stores. When Pseudomonas aeruginosa bacteria obtained from the Indonesian Institute of Sciences (LIPI) at IPB University, Indonesia (Fungai) arrive in the laboratory, the standard number of bacteria added by McFarland (0.5) is equivalent to 1.5 x 10 8 bacterial cells/ml. 2.2 Preparation of Pseudomonas aeruginosa bacteria Pseudomonas aeruginosa bacteria were obtained from the Indonesian Institute of Sciences (LIPI) at IPB University, Indonesia. According to (Palareti et al., 2016 ), bacteria in the genus Pseudomonas aeruginosa are a good bioremediation agent for hydrophobic polymers due to the attachment of bacterial cells to the surface of the polymer, which many bacteria do not have. These bacteria have many catalytic enzymes and pathways for very high metabolism. specific area of plastic polymer. Thus, the presence of these bacteria will make Polypropylene (PP) more hydrophilic so as to facilitate microbial attack for further degradation processes (Habib et al., 2020 ). Bacterial cultures obtained in test tube containers were taken using sterile wire loops and inoculated with the culture method on Nutrient Agar (NA) media, MERCK 20 gram/L, in a petri dish. Bacteria were incubated for 7 days on Nutrient Agar (NA) at 37°C. After the bacteria are ready, the bacteria in solid media are diluted before being applied with the 0.5 McFarland standard, which is commonly used as a turbidity comparison in bacterial cultures in liquid medium, where the density of bacteria in the liquid medium is 1 x 10 7 – 1 x 10 8 cells/ml. The preparation of a standard 0.5 McFarland solution was carried out by sterilizing all equipment using an autoclave at 121°C for 15 minutes. A test tube was then filled with 9 ml of sterile distilled water under aseptic conditions. Pseudomonas aeruginosa from solid media was transferred in a zig-zag manner into the test tube containing sterile distilled water using a disposable 6-inch cotton swab. The suspension was mixed until homogeneous, and the turbidity was adjusted to the 0.5 McFarland standard (1 × 10⁸ cells/ml). The prepared suspension was subsequently applied to the soil. 2.3 Photo-oxidative degradation of disposable mask using UV/H 2 O 2 The sample preparation stage before treatment is an important step. SENSI surgical face mask, a 3-ply earloop used as many as 10 sheets, which were removed, then the UV/H 2 O 2 process was carried out with a composition of hydrogen peroxide H 2 O 2 12% V/V assisted by irradiating UV-C lamps at a wavelength of 248–262 nm. According to (Beltrán et al., 2024 ) UV at a wavelength of 180/254 nm produces significant shape changes in the form of cracks and stretching. The UV/H 2 O 2 experiment was carried out for 30 hours at room temperature (25°C) to speed up the mask degradation process. After going through these stages, the mask is allowed to air dry at room temperature (25°C–30°C) for 1 day until it is dry. Next, the mask is cut into four parts and tied end to end with thread to be inserted in the middle of the soil treatment with the aim of facilitating mask sampling. Mask samples without treatment were also used in this study, as many as four sheets with the same treatment were used. The average thickness of the control mask for the front, middle, and back layers measured separately was 0.102 ± 0.010 mm; 0.108 ± 0.010mm and 0.086 ± 0.007mm were measured using a Digital micrometer of 0.001 mm, Syntek 0–25 mm (China). 2.4 Biodegradation of photo-oxidized mask The biodegradation process was conducted for two months with two collections, in the first and the second month. Two masks were taken for each collection and then placed in a clip container for storage and analysis. The variables used were 5 pots with each name, namely P1 (photo-oxidized mask + no bacteria + no worms), P2 (photo-oxidized mask + bacteria + no worms), P3 (photo-oxidized mask + bacteria + worms), P4 (untreated mask + bacteria + worms), and P5 (untreated mask + no bacteria + no worms). A sample of the SENSI surgical face mask, a 3-ply earloop that has been conditioned with a string, is inserted into half the depth of the pot, and as many as four masks are spread around the plants. Each pot is prepared so that the soil stays moist and the plants receive regular waterings to remain alive. The medium in this study was garden soil (loamy soil), which had been conditioned in the pot according to the variables. Garden soil was taken at different points at depth (0–15 cm), which was then homogenized as soil without treatment and with the addition of Trubus compost in a ratio of 3:1 as treated soil. The addition of worms, namely 18 Lumbricus rubellus with uniform sizes, chili plants of the Capsicum annuum L type used were ± 20 cm high, and 0.5 McFarland standard Pseudomonas aeruginosa bacteria. Bacteria are added above the soil surface by as much as 0.5 McFarland standard as a result of changes in mask degradation. The research treatment was placed outdoors with sunlight settings using black nets (paranet). 2.5 Microscopy analysis The treatment of mask analysis using a microscope needs to be prepared by cleaning particles in the form of dirt and solution attached to the mask so that there are no contaminants in the mask under the microscope. The samples analyzed were mask samples without treatment, photo-oxidized mask samples and treated samples in the soil. The protocol was carried out by first washing the mask sample thoroughly with 96% (v/v) ethanol (14.44 M) and stirring at 600 rpm for ± 15 minutes or until the sample is clean. Samples that had been added to ethanol were washed with distilled water for 3 minutes and allowed to stand for 1 minute to separate the organic matter (Herrera et al., 2021 ). If there are still particles or dirt attached to the mask, carefully sort them out using tweezers. Clean mask samples were air-dried for 24 hours to facilitate analysis using a microscope. After going through the protocol, the masks were visually seen under the Stereo Zoom NSZ 60 Microscope, Opti lab Advance Plus were photographed before and after treatment at 2X magnification and 5X maximum magnification. Opti lab is an application used to run a microscope camera. Then the images obtained from the Opti lab application were analyzed using the Image Raster application. A scanning electron microscope (SEM) analysis was also carried out to see changes in the morphology of the mask surface (Arkatkar et al., 2010 ). SEM analysis was performed at 1000x magnification, 3000x, 5000x, and 10,000x. SEM analysis was carried out on samples of masks without treatment, photo-oxidized masks, and masks after treatment in the 2nd month to see changes in the surface of the masks before and after degradation in ISO 846 and ISO 11266, which were indicated by the colonization of microorganisms on the sample surface (Lucas et al., 2008 ). SEM images will provide a visual form showing that masks have smooth surface shapes and mask fibers that are tight or stretchy (Gomes de Aragão Belé et al., 2021). After exposure to photoocsidation and mask treatment, the soil will show stretching or cracks in the fiber structure. 2.6 FTIR analysis Fourier Transform infrared Spectroscopy (FTIR) analysis was performed to see changes in structural and functional groups on the polypropylene mask pieces. FTIR analysis was carried out on untreated masks, photo-oxidized masks, and treated masks using a Perkin Elmer Type Frontier instrument capable of producing a spectrum of 4000 − 500 cm-1 (SNI 19-4370-2004 method) and ASTM D6288-89. Fourier transform infrared Spectroscopy (FTIR) spectroscopy will reveal chemical bonds (functional groups). The identification that is easily provided through FTIR analysis is a carbon-based polymer. Information via FTIR tells the state of the oxygen bonding components (e.g., carbonyl groups) and hydroxyl groups in the polypropylene material from mask oxidation (6). The determination of the normal intensity of a functional group is based on transmittance (%) (Habib et al., 2020 ). Result and Discussion 3.1 Physical characteristics of photo-oxidized and biodegraded mask A 3-layer disposable mask, each of which has different material characteristics: the outer layer is made of spun bond polypropylene, the middle layer is made of Melt-blown polypropylene, and the inner layer is made of a mixture of polypropylene with polyester or cotton will have a different degradation process description with the same treatment. The photooxidation treatment of disposable masks causes significant cracks on their surface, leading to brittleness of the material. These cracks result from the breakdown of polymer chains reacting with hydroxyl radicals (Liu et al., 2022). These cracks can reduce the elasticity of the mask fibers, causing the photo oxidized mask fibers to look more tenuous than those without oxidation (Tsubone et al., 2019). The photooxidation treatment also accelerated the degradation of the masks in soil compared to the masks that were not subjected to similar treatment. This can be observed in Fig. 1 (pots 1, 2, and 3), which show more tenuous mask fibers and more biofilm build-up compared to pots 4 and 5 that were not photo oxidized. More biofilm formation on oxidized mask surfaces occurs because photooxidation accelerates the biodegradation process. Photooxidation causes mechanical fragmentation of the mask material, producing small fragments that provide more surface for soil microbes to break down the mask polymer (Rizzarelli et al., 2021). In addition, the addition of Pseudomonas aeruginosa bacteria and earthworms led to increased biofilm production. Pseudomonas aeruginosa bacteria can trigger the rate of bacterial activity in the soil. It can be seen in pots 3 and 4 that the addition of biofilm increased significantly which was marked by the presence of more and more yellow color in the 2nd layer. Pseudomonas aeruginosa bacteria are known to be able to degrade various organic and inorganic compounds, and produce enzymes such as proteases and lipases that play a role in breaking down the molecular structure of mask polymers, thus supporting the degradation process (Mohanan et al., 2020). The addition of earthworms in the soil system, as shown in pot 3 and pot 4, plays a role in accelerating the biodegradation process carried out by bacteria. Earthworms break down organic matter into smaller particles through mechanical digestion (Wang et al., 2021), such digestion results in nutrient-rich excretions often referred to as vermicast. Vermicast contains organic substances and beneficial microbes that can stimulate the formation of bacteria, favorable bacterial growth resulting in a significant increase in biofilm compared to pot 5 and pot 2. In addition, the burrowing activity performed by earthworms also increases soil aeration and moisture, creating more conducive conditions for decomposing microorganisms (Ahmed and Al-Mutairi, 2022). Based on Table 1, the change in thickness of each layer of the disposable mask is seen to decrease significantly due to the photooxidation treatment compared to the control. The photooxidation process involves a reaction between the polypropylene in the mask and oxygen in the air, triggered by exposure to ultraviolet (UV) light from the sun. UV radiation breaks the polymer chains present in polypropylene, generating free radicals, this is a precursor to the degradation of polymer molecules into smaller molecular fragments, such as hydroperoxides and carbonyls. These fragments cause brittleness of the material, so the mask layer begins to crack (Tang et al., 2019). Biodegradation usually consists of three distinct steps: 1) formation of biofilm formation on the plastic surface; 2) decomposition of the plastic into smaller molecules through the action of extracellular enzymes secreted by the microorganisms; and 3) consumption and further metabolism of these smaller molecules these smaller molecules within the cell (Andrady et al., 2022a). The thickness test was conducted to see the change in thickness of each mask layer after the degradation process. Ultraviolet radiation in the photooxidation process on disposable mask polymers results in damage to the polymer chain which has an impact on the deterioration of plastic mechanical properties including the change of thickness (Wang et al., 2021) . Table 1. Thickness analysis of disposable mask. Data are shown as mean ± SD (n = 2). In table.1 . shows a significant decrease in thickness in all mask layers after photooxidation treatment. The decrease also occurred after the disposable masks were buried in the ground, both with the addition of bacteria and without the addition of bacteria. Meanwhile, the masks with photooxidation treatment and continued burial in soil with the addition of bacteria (P2 and P3) showed a greater decrease in thickness when compared to disposable masks without photooxidation both with burial in soil using bacteria and without bacteria. However, photodegradation is an important process in plastic degradation that causes changes in plastic thickness and structural integrity (Sivasankar and Sunitha, 2024). After the photooxidation lasted longer, the cracks on the surface of the mask began to deepen and spread to the inside of the mask, reducing the thickness of the mask gradually (Table 2). Macroscopically, the thickness of the mask will decrease as the material structure is destroyed. In addition, the addition of bacteria and earthworms to the biodegradation process in the soil also causes a decrease in thickness in each layer. Bacteria release extracellular enzymes that break down the polymer chains in the mask. The broken polymer chains cause the mask to lose thickness (Mohanan et al., 2020). Meanwhile, the addition of earthworms plays a role in accelerating the biodegradation process by providing a physical and chemical environment where decomposing microbes thrive such as Pseudomonas aeruginosa bacteria (Ahmed and Al-Mutairi, 2022). Seen in P4, the addition of bacteria and earthworms resulted in a decrease in thickness compared to P5, without the addition of bacteria and earthworms. Table 2 Thickness analysis of disposable mask. Data are shown as mean ± SD (n = 2). Layer Sample Untreated Mask Photo-oxidized Mask P1 Mask P2 Mask P3 Mask P4 Mask P5 Mask 1st Layer 0.102 ± 0.010 0.077 ± 0.007 0.070 ± 0.009 0.069 ± 0.006 0.070 ± 0.005 0.081 ± 0.001 0.087 ± 0.001 2nd Layer 0.108 ± 0.010 0.087 ± 0.006 0.079 ± 0.001 0.081 ± 0.001 0.079 ± 0.002 0.089 ± 0.001 0.090 ± 0.002 3rd Layer 0.086 ± 0.007 0.067 ± 0.008 0.061 ± 0.002 0.065 ± 0.001 0.061 ± 0.002 0.067 ± 0.002 0.069 ± 0.002 3.2 FTIR analysis of photo-oxidized and biodegraded mask Oxidative photodegradation (photo-oxidation) of polypropylene is an important stage before reaching the biodegradation stage, due to the fact that biodegradation process in plastics occurs when carbonyl groups have been formed on polymers (Andrady et al., 2022a). In the research of Khoironi et al., (2020), the photo-oxo degradation process is able to provide significant changes in the molecular structure of organic polypropylene with the formation of new groups that are very important in the attack of microorganisms. The formation of carbonyl groups such as carboxylic acids, aldehydes and ketones will increase the moisture (hydrophilicity) of polypropylene, increase amorphous properties, form pores on the plastic surface due to holes and cracks (Esmaeili et al., 2013). Of course, these conditions will facilitate the attack of microorganisms that will work in biodegradation (Mohanan et al., 2020). Photo-oxidation of polypropylene disposable masks produces carbonyl, chain unsaturation and carboxylic acid products, which will initiate the next process where weathering due to this attack will increase the ability to absorb ultraviolet light photo-initiation (Andrady et al., 2022b). As a consequence of the photo-oxidation process, there is a great influence on the physical and mechanical properties of polypropylene in disposable masks. In the oxygen diffusion controlled photo-oxidation process, crystallinity will also increase with the oxidation rate. The dismemberment of macromolecules involves the formation of smaller molecules that allow easier crystallization. Furthermore, this change will lead to a decrease in molar mass caused by chain cutting due to UV exposure for a long time, thus changing from ductile to brittle behavior which results in easy attack by microorganisms in the biodegradation process (Yousif and Haddad, 2013). From Fig. 2. Shows changes in all layers of disposable masks. These changes regarding the formation of new functional groups indicate the uptake of oxygen due to surface vulnerability as a result of ultra-violet radiation. In the first layer of the mask after being treated with photo-oxidation, there are several peaks formed at waves that are not present in the control mask, each of which is 899 cm-3 which is aimed at ketones, then 1102 cm -1 and 1256 cm -3 which both point to carboxylic acids, then 1302 cm -3 which points to esters and 2838 cm -3 at the -OH group. Similar to what happened in the first layer, some new peaks also appeared in the second layer of disposable masks after photo-oxidation treatment. The appearance of new peaks is indicated by peaks at a wavelength of 899 cm -3 which is aimed at ketones then 1102 cm -3 and 1256 cm -3 which both lead to carboxylic acids and 1302 cm-3 which leads to esters, this also appears in the third layer except 1302 cm -3 which is not visible in the third layer of disposable masks after photo-oxidation treatment According to Zhu et al., (2020), cycles in photo-oxidation will facilitate the attack of microorganisms to produce rapid fragmentation which will lead to complete mineralization in the biodegradation process. The presence of worms and pseudomona s bacteria in this study did not appear to have a significant impact after the disposable masks were buried in the soil for a long time. In 2nd layer disposable masks (Fig. 2B) with photo-oxidation treatment (P2, P3), there is a peak at 1302 cm-3 that leads to esters and a peak at 3393 cm-3 that leads to ketones, these two peaks have the same intensity even though the two reactors (P2 and P3) were treated with different organism interventions, it appears that the presence of pseudomonas and worms does not have an impact on the presence of worms. on the other hand, a peak appears to appear in the FTIR analysis results for masks without photo-oxidation (P4 and P5) at a wavelength of 1302 cm -3 which is directed at the ketone functional group. In the first layer of disposable masks new peaks appeared only on the masks in reactors P2, P3 and P4 where the reactors received organism treatment, although it seems that organisms do not really play a role in this degradation process but the appearance of peaks at important wavelengths produced by the masks in the reactor indicates the role of the organism. The peaks that appear are at a wavelength of 1164 cm − 3 which leads to carboxylic acids and a wave of 1654 cm − 3 for P2 which leads to the formation of ketones and a wavelength of 1647 cm − 3 in the three reactors which leads to carbonyl groups. In the third layer of disposable masks, both masks that have undergone photo-oxidation and those without photo-oxidation treatment do not seem to provide significant results after being buried in the ground. There are no significant peaks that indicate changes in functional groups due to biological or abiotic attacks in the soil, except for the appearance of one peak at a wavelength of 3695 cm- 3 which leads to hydroxyl and carbonyl groups that appear on the mask at P2, P3 and P4. 3.3 Morphological analysis using scanning electron microscopy To evaluate the impact of photooxidation on biodegradation efficacy, masks were buried for 100 days, including both treated and untreated masks. Figure 3. illustrates alterations in the morphology of the mask fibers both untreated and photo- oxidized masks post-burial. The changes encompass the formation of biofilms, the emergence of cracks on the mask fibers' surface, and an increase in mask porosity. This may arise from microbial activity, chemical reactions, and physical degradation. Polypropylene mask fibers can serve as a substrate for biofilm development due to their properties. Hydrocarbon-degrading bacteria attaching to the mask's surface generate enzymes that cleave the long polymer chains in polypropylene into smaller units, thus altering the physical properties of the mask 2425. Figure 3. shows uneven surfaces, cracks and loosen fibers in photo-oxidized masks, which were not appeared in non-photo-oxidized masks. This results align with a study by 6, which reported that disposable masks began to degrade after being exposed to UV irradiation for 36 hour. In current study, the photooxidation process was conducted for 72 hours and assisted by an oxidizing agent (H 2 O 2 ). When H 2 O 2 is exposed to UV light, it breaks down into two hydroxyl radicals (•OH) .These radicals attack the polymer chains of the mask, which over time, can cause the mask to become brittle and loosen its fibers 6 The fine cracks formed through photooxidation cause the mask fiber's surface rougher. The rough texture of the mask fiber increases the surface area for bacterial adhesion, while the irregularities on the rough surface shield the bacteria, safeguarding them from being displaced by physical disruptions such as water flow from irrigation (Mu et al., 2023). Research indicates that bacteria, including Escherichia coli, Pseudomonas aeruginosa , and Staphylococcus aureus , adhere and move more effectively to rough polypropylene fibers compared to smooth ones. The increased movement and adherence of bacteria result in the formation of more biofilms (Cai et al., 2019). In this investigation, the biofilm developed in P3 exhibited no significant difference when compared to P5. This may be affected by soil conditions that facilitate the enzymatic activities of Pseudomonas aeruginosa on masks, regardless of photooxidation treatment on masks. Furthermore, in the control mask, the disparity in surface roughness of the mask fibers with and without photooxidation was minimal, resulting in comparable bacterial activity on both treatment. Consequently, additional research is required to determine the optimal duration for the photooxidation process of disposable masks. The incorporation of Pseudomonas aeruginosa into the growth medium augmented biofilm production, as illustrated in Fig. 3, where P2, P3, and P4 exhibit more biofilm accumulation compared to reactors P5 and P1. The introduction of Pseudomonas aeruginosa serves as an inoculant, stimulating the rate of bacterial activity in the soil. The incorporation of worms into the growth media enhances the quantity of biofilm produced, as worms optimize the physical and chemical conditions conducive to microbial proliferation. Worms make castings (excrement) that are abundant in organic nutrients and create an optimal environment for bacterial proliferation. These castings possess an increased surface area and comprise water, nutrients, and microbes, which enhance microbial activity. Furthermore, earthworms enhance soil aeration and water retention via their burrows, fostering an environment with elevated oxygen and water availability, both crucial for bacterial metabolism (Buivydaitė et al., 2023; Duan et al., 2024; Zhang et al., 2025). Disposable surgical masks commonly comprise three layers, each constructed from distinct materials with certain characteristics that enhance the mask's overall efficacy. The outer layer consists of spun-bond polypropylene and possesses hydrophobic properties, which reject water and inhibit the ingress of droplets and larger particles into the mask (Res et al., 2018). The second layer consists of melt-blown polypropylene, formed from ultra-fine fibers that generate a dense, electrostatically charged structure. The third layer of disposable surgical masks comprises spun-bond polypropylene blended with soft polyester or cotton (J. Wang et al., 2023). Figure 3. illustrates that biofilm accumulation is more pronounced in the middle layer than in the other layers. This may result from the material composition and structure of the melt-blown polypropylene utilized in this layer. The melt-blown polypropylene layer possesses a thin and porous structure that more effectively captures particles and retains moisture compared to the outer and inner layers. The accumulated moisture fosters an environment that promotes bacterial proliferation. The increased surface area of the intermediate layer offers more areas for bacterial attachment and biofilm formation. In contrast to the hydrophobic outer layer that repels moisture, the inner layer's composition enables it to retain moisture and organic matter, including soil nutrients, so facilitating bacterial colonization. The confined particles and moisture create an environment and nutrient source for bacteria, hence improving biofilm formation. The high surface area, moisture retention, and porous characteristics of the middle layer render it more prone to bacterial colonization and biofilm formation than the other mask layers. This aligns with study (Jeong et al., 2021), which shown that bacterial activity in the middle layer exceeded that of the outer and inner layers. Conclusion From the data obtained in this study, there is a tendency to have the same phenomenon, where photo-oxidation treatment on disposable mask samples does not provide significant changes after the mask is buried in the ground. Mechanical changes using tests using a microscope, thickness and SEM show the influence of pretreatment on masks that experience photo-oxidation. Significantly, the appearance of biofilm is accompanied by the formation of holes and an increase in pores on the surface of the mask which plays a role in mask fragility. Biological attack also causes the mask fibers to be more tenuous, this shows the increased hydrophilic properties of the mask, it seems that the moisture generated by the biofilm plays a big role in this. The mechanical changes that occur are of course supported by changes in the molecular structure of the mask, where the FTIR test results show the emergence of important functional groups that play a role in the degradation process, namely carboxylates, ketones and esters in Reactor R3, R4 and R5. To find out more precisely about the involvement of organisms in achieving complete biodegradation results, it is necessary to carry out a longer burial process by monitoring the life of the organisms involved and the stability of temperature and humidity. Declarations Conflict of Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Ethical Declaration This study was conducted in accordance with the applicable institutional and national guidelines. The photo-oxidation procedure adhered to ASTM D5208 (Standard Practice for Fluorescent Ultraviolet Exposure of Photodegradable Plastics). Plant physiological responses were assessed using standard, widely accepted physiological evaluation methods, as no specific guideline exists for this type of plant assay. Ethical approval was not required for this research because it did not involve human participants, personal data, or vertebrate animals. Consent to Participate Not applicable. No human participants were involved in this study. Consent to Publish Not applicable. This manuscript does not contain any individual person’s data in any form (including images, videos, or identifiable information). Fundings This research was financially supported by The World Class University Research (RWCU) by Diponegoro University, Grant-No: 222–771/UN7.D2/PP/IV/2025 and KAKENHI (24K00992). Author Contribution Conceptualization: Hadiyanto, Adian Khoironi; Methodology: Hadiyanto, Adian Khoironi, Inggar Dianratri; Investigation and Data Curation: Falvocha Alifsmara Joelyna, Rifqi Ahmad Baihaqi, Wahyu Zuli Pratiwi, Wahyu Diski Pratama; Formal Analysis: Falvocha Alifsmara Joelyna; Validation: Widayat, Marcelinus Christwardana, Adhelia Intan Sabhira; Resources and Supervision: Hadiyanto, Adian Khoironi, Widayat; Writing Original Draft: Adian Khoironi; Writing Review & Editing: Adian Khoironi, Marcelinus Christwardana, Hadiyanto, Tomoya Kataoka; Project Administration and Funding Acquisition: Hadiyanto, Adian Khoironi Acknowledgement The authors would like to express their profound gratitude to the laboratory assistants at the Center of Biomass and Renewable Energy Laboratory for their essential technical support for the entirety of this research endeavor. Their proficiency and assistance have been vital in the accomplished implementation of this project. 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1","display":"","copyAsset":false,"role":"figure","size":93124,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectrum of untreated and treated mask using photo-oxidation, A) 1\u003csup\u003est\u003c/sup\u003e layer, B) 2\u003csup\u003end\u003c/sup\u003e layer, and C) 3\u003csup\u003erd\u003c/sup\u003e layer.\u003c/p\u003e","description":"","filename":"Picture1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8082479/v1/77b52facbc4f52f51d7b0f72.jpg"},{"id":97142560,"identity":"86b095ce-ed86-452b-9033-1bd5bc3630dc","added_by":"auto","created_at":"2025-12-01 10:07:43","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":78728,"visible":true,"origin":"","legend":"\u003cp\u003eFTIR spectrum of photo-oxidized mask and biodegraded mask in different conditions, A) 1\u003csup\u003est\u003c/sup\u003e layer, B) 2\u003csup\u003end\u003c/sup\u003e layer, and C) 3\u003csup\u003erd\u003c/sup\u003e layer.\u003c/p\u003e","description":"","filename":"Picture2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8082479/v1/fea2bc8e4dcdd6a53b0e8fa8.jpg"},{"id":97133280,"identity":"1a96c89b-8d2f-4595-abf6-d0041a569625","added_by":"auto","created_at":"2025-12-01 09:07:20","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1201337,"visible":true,"origin":"","legend":"\u003cp\u003eSEM image of mask.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8082479/v1/4b6eb7d6d1985ba7005f9046.png"},{"id":105892733,"identity":"b367017c-c1da-427c-b66b-b006bb2aeb50","added_by":"auto","created_at":"2026-04-01 08:14:15","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2968797,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8082479/v1/6b4f7061-6c86-4e50-8d26-145dd2b475cd.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Photo-oxidative degradation of disposable mask and the effect on the biodegradation process in soil system","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe surge in disposable mask waste can be attributed to the heightened demand resulting from the COVID-19 pandemic. The production of masks has escalated in response to the growing demand, which is closely correlated with daily usage. Face masks are utilized to safeguard Public Health against the transmission of diseases. The global monthly usage of masks has surged to 129\u0026nbsp;billion, with projections indicating a further upward trend (Wang et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). On a daily basis, individuals dispose of around 15.5 trillion tons of mask garbage, which corresponds to 2\u0026nbsp;million tons of plastic waste. Each mask weighs around 15.5 grams (Wang et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Hence, the surge in discarded face masks has emerged as a worldwide contributor to plastic contamination. The daily quantity of disposable mask garbage in Indonesia ranges from 53,840 to 80,759\u0026nbsp;million pieces. Approximately 10\u0026nbsp;million masks are projected to be discarded on a monthly basis, with an expected total of 1.56\u0026nbsp;billion masks being disposed of in the year 2020.\u003c/p\u003e\u003cp\u003eDisposable masks mostly comprise polypropylene, polyacrylonitrile, and polystyrene polymers (Akber Abbasi et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Multiple studies have proven that mask waste is a contributing factor to plastic pollution. The COVID-19 epidemic has worsened the problem of plastic pollution, with thousands of discarded masks being disposed of in the environment. This has resulted in potential toxicological risks, particularly for marine environments (Luna et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Mask manufacture involves the use of chemicals that can undergo decomposition or degradation as a result of physical, chemical, or biological interactions with the natural environment. This particular variant of a three-layer disposable mask offers antibacterial protection, effectively mitigating the transmission of diseases through droplets emitted by individuals in close proximity. This mask serves multiple functions, including filtration, moisture absorption, and water resistance. According to (Farzaneh and Shirinbayan, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), its effectiveness has been demonstrated and is claimed to be comparable to that of a N95 respirator. However, it is unable to block extremely minute droplets. A disposable mask is composed of three layers: an inner layer that is hydroscopic, a middle layer that acts as a fine filter, and a back layer that is water resistant. The mask consists of three layers: the outer layer is composed of spun-bonded non-woven fabric, the middle layer is made of melt-blown non-woven fabric, and the inner layer is also spun-bonded non-woven fabric (J. Wang et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Non-woven materials, like polypropylene, possess notable attributes such as high temperature tolerance and a structurally stable composition. In addition, non-woven materials are single-use only. The intermolecular connections between the fibers of the mask are strengthened as a result of the introduction of exogenous substances.\u003c/p\u003e\u003cp\u003eDisposable masks can undergo biodegradation in the soil, facilitated by the actions of living organisms (Lucas et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Nevertheless, disposable masks are typically composed of polypropylene or polyethylene, both of which are robust polymers that exhibit limited degradability. This can result in an extended duration for the natural degradation process, potentially exceeding a decade (Canopoli et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Consequently, multiple techniques have been devised to accelerate this process of degradation. Liu et al., (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) conducted an experiment to assess the properties of disposable masks following exposure to UV radiation. The results revealed a reduction in elongation at break, tensile strength, and maximum force of the masks, suggesting the occurrence of chemical bond breakdown. A recent study conducted by Y. Wang et al., (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) employed the oxo degradation technique, utilizing O\u003csub\u003e3\u003c/sub\u003e and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, to facilitate the degradation of polyethylene. The investigation revealed alterations in the polymer bonds, rendering them more susceptible to breakage. The combination of UV radiation with oxidation chemicals, such as hydrogen peroxide, can generate hydroxyl radicals. These radicals facilitate the degradation of the polymer bonds. Easton et al., (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) documented the development of superficial cavities, depressions, and cracks on the surface of the microplastic, as well as alterations in the proportion of oxygen-containing functional groups, following its exposure to UV/ H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e treatment. The presence of cavities, cracks, and alterations in polymer linkages that arise during the photo-oxo degradation process will facilitate the acceleration of the biodegradation process. The objective of this study is to assess the impact of photo-oxo degradation on the biodegradation process inside the soil system. In order to replicate a natural biodegradation environment, single-use masks were placed in a pot along with \u003cem\u003epseudomona\u003c/em\u003es bacteria, earthworms, and plants for a duration of over 100 days. The masks underwent evaluation of their physical and chemical alterations following treatment by the use of SEM, Microscope, micrometer screw gauge, and FTIR analysis. By integrating the photo-oxo degradation and biodegradation mechanisms, we aim to address the escalating issue of mask waste, which has been exacerbated by the COVID-19 pandemic.\u003c/p\u003e"},{"header":"Materials and Method","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003e2.1 Materials\u003c/h2\u003e\u003cp\u003eThe sample used is the Sensi mask of a surgical face mask, a 3-ply earloop, SENSI brand, obtained from Arista Latino Company, Indonesia, standardized for Disposable Mask Characteristics (SNI EN 14683:2019; AC: 2019; and SNI 8488:2018). The soil medium used is loamy soil. The compost used was Trubus compost obtained from a plant store. Additional worms used were Lumbricus rubellus and Capsicum annuum L plants obtained from plant stores. When \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e bacteria obtained from the Indonesian Institute of Sciences (LIPI) at IPB University, Indonesia (Fungai) arrive in the laboratory, the standard number of bacteria added by McFarland (0.5) is equivalent to 1.5 x 10\u003csup\u003e8\u003c/sup\u003e bacterial cells/ml.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003e2.2 Preparation of \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e bacteria\u003c/h2\u003e\u003cp\u003ePseudomonas aeruginosa bacteria were obtained from the Indonesian Institute of Sciences (LIPI) at IPB University, Indonesia. According to (Palareti et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), bacteria in the genus Pseudomonas aeruginosa are a good bioremediation agent for hydrophobic polymers due to the attachment of bacterial cells to the surface of the polymer, which many bacteria do not have. These bacteria have many catalytic enzymes and pathways for very high metabolism. specific area of plastic polymer. Thus, the presence of these bacteria will make Polypropylene (PP) more hydrophilic so as to facilitate microbial attack for further degradation processes (Habib et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Bacterial cultures obtained in test tube containers were taken using sterile wire loops and inoculated with the culture method on Nutrient Agar (NA) media, MERCK 20 gram/L, in a petri dish. Bacteria were incubated for 7 days on Nutrient Agar (NA) at 37\u0026deg;C. After the bacteria are ready, the bacteria in solid media are diluted before being applied with the 0.5 McFarland standard, which is commonly used as a turbidity comparison in bacterial cultures in liquid medium, where the density of bacteria in the liquid medium is 1 x 10\u003csup\u003e7\u003c/sup\u003e \u0026ndash; 1 x 10\u003csup\u003e8\u003c/sup\u003e cells/ml. The preparation of a standard 0.5 McFarland solution was carried out by sterilizing all equipment using an autoclave at 121\u0026deg;C for 15 minutes. A test tube was then filled with 9 ml of sterile distilled water under aseptic conditions. \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e from solid media was transferred in a zig-zag manner into the test tube containing sterile distilled water using a disposable 6-inch cotton swab. The suspension was mixed until homogeneous, and the turbidity was adjusted to the 0.5 McFarland standard (1 \u0026times; 10⁸ cells/ml). The prepared suspension was subsequently applied to the soil.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003e2.3 Photo-oxidative degradation of disposable mask using UV/H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/h2\u003e\u003cp\u003eThe sample preparation stage before treatment is an important step. SENSI surgical face mask, a 3-ply earloop used as many as 10 sheets, which were removed, then the UV/H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e process was carried out with a composition of hydrogen peroxide H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e 12% V/V assisted by irradiating UV-C lamps at a wavelength of 248\u0026ndash;262 nm. According to (Beltr\u0026aacute;n et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) UV at a wavelength of 180/254 nm produces significant shape changes in the form of cracks and stretching. The UV/H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e experiment was carried out for 30 hours at room temperature (25\u0026deg;C) to speed up the mask degradation process. After going through these stages, the mask is allowed to air dry at room temperature (25\u0026deg;C\u0026ndash;30\u0026deg;C) for 1 day until it is dry. Next, the mask is cut into four parts and tied end to end with thread to be inserted in the middle of the soil treatment with the aim of facilitating mask sampling. Mask samples without treatment were also used in this study, as many as four sheets with the same treatment were used. The average thickness of the control mask for the front, middle, and back layers measured separately was 0.102\u0026thinsp;\u0026plusmn;\u0026thinsp;0.010 mm; 0.108\u0026thinsp;\u0026plusmn;\u0026thinsp;0.010mm and 0.086\u0026thinsp;\u0026plusmn;\u0026thinsp;0.007mm were measured using a Digital micrometer of 0.001 mm, Syntek 0\u0026ndash;25 mm (China).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\u003ch2\u003e2.4 Biodegradation of photo-oxidized mask\u003c/h2\u003e\u003cp\u003eThe biodegradation process was conducted for two months with two collections, in the first and the second month. Two masks were taken for each collection and then placed in a clip container for storage and analysis. The variables used were 5 pots with each name, namely P1 (photo-oxidized mask\u0026thinsp;+\u0026thinsp;no bacteria\u0026thinsp;+\u0026thinsp;no worms), P2 (photo-oxidized mask\u0026thinsp;+\u0026thinsp;bacteria\u0026thinsp;+\u0026thinsp;no worms), P3 (photo-oxidized mask\u0026thinsp;+\u0026thinsp;bacteria\u0026thinsp;+\u0026thinsp;worms), P4 (untreated mask\u0026thinsp;+\u0026thinsp;bacteria\u0026thinsp;+\u0026thinsp;worms), and P5 (untreated mask\u0026thinsp;+\u0026thinsp;no bacteria\u0026thinsp;+\u0026thinsp;no worms). A sample of the SENSI surgical face mask, a 3-ply earloop that has been conditioned with a string, is inserted into half the depth of the pot, and as many as four masks are spread around the plants. Each pot is prepared so that the soil stays moist and the plants receive regular waterings to remain alive. The medium in this study was garden soil (loamy soil), which had been conditioned in the pot according to the variables. Garden soil was taken at different points at depth (0\u0026ndash;15 cm), which was then homogenized as soil without treatment and with the addition of Trubus compost in a ratio of 3:1 as treated soil. The addition of worms, namely 18 Lumbricus rubellus with uniform sizes, chili plants of the Capsicum annuum L type used were \u0026plusmn;\u0026thinsp;20 cm high, and 0.5 McFarland standard \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e bacteria. Bacteria are added above the soil surface by as much as 0.5 McFarland standard as a result of changes in mask degradation. The research treatment was placed outdoors with sunlight settings using black nets (paranet).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\u003ch2\u003e2.5 Microscopy analysis\u003c/h2\u003e\u003cp\u003eThe treatment of mask analysis using a microscope needs to be prepared by cleaning particles in the form of dirt and solution attached to the mask so that there are no contaminants in the mask under the microscope. The samples analyzed were mask samples without treatment, photo-oxidized mask samples and treated samples in the soil. The protocol was carried out by first washing the mask sample thoroughly with 96% (v/v) ethanol (14.44 M) and stirring at 600 rpm for \u0026plusmn;\u0026thinsp;15 minutes or until the sample is clean. Samples that had been added to ethanol were washed with distilled water for 3 minutes and allowed to stand for 1 minute to separate the organic matter (Herrera et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). If there are still particles or dirt attached to the mask, carefully sort them out using tweezers. Clean mask samples were air-dried for 24 hours to facilitate analysis using a microscope. After going through the protocol, the masks were visually seen under the Stereo Zoom NSZ 60 Microscope, Opti lab Advance Plus were photographed before and after treatment at 2X magnification and 5X maximum magnification. Opti lab is an application used to run a microscope camera. Then the images obtained from the Opti lab application were analyzed using the Image Raster application.\u003c/p\u003e\u003cp\u003eA scanning electron microscope (SEM) analysis was also carried out to see changes in the morphology of the mask surface (Arkatkar et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). SEM analysis was performed at 1000x magnification, 3000x, 5000x, and 10,000x. SEM analysis was carried out on samples of masks without treatment, photo-oxidized masks, and masks after treatment in the 2nd month to see changes in the surface of the masks before and after degradation in ISO 846 and ISO 11266, which were indicated by the colonization of microorganisms on the sample surface (Lucas et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). SEM images will provide a visual form showing that masks have smooth surface shapes and mask fibers that are tight or stretchy (Gomes de Arag\u0026atilde;o Bel\u0026eacute; et al., 2021). After exposure to photoocsidation and mask treatment, the soil will show stretching or cracks in the fiber structure.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003e2.6 FTIR analysis\u003c/h2\u003e\u003cp\u003eFourier Transform infrared Spectroscopy (FTIR) analysis was performed to see changes in structural and functional groups on the polypropylene mask pieces. FTIR analysis was carried out on untreated masks, photo-oxidized masks, and treated masks using a Perkin Elmer Type Frontier instrument capable of producing a spectrum of 4000\u0026thinsp;\u0026minus;\u0026thinsp;500 cm-1 (SNI 19-4370-2004 method) and ASTM D6288-89. Fourier transform infrared Spectroscopy (FTIR) spectroscopy will reveal chemical bonds (functional groups). The identification that is easily provided through FTIR analysis is a carbon-based polymer. Information via FTIR tells the state of the oxygen bonding components (e.g., carbonyl groups) and hydroxyl groups in the polypropylene material from mask oxidation (6). The determination of the normal intensity of a functional group is based on transmittance (%) (Habib et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e"},{"header":"Result and Discussion","content":"\u003cdiv id=\"Sec10\"\u003e\n \u003ch2\u003e3.1 Physical characteristics of photo-oxidized and biodegraded mask\u003c/h2\u003e\n \u003cp\u003eA 3-layer disposable mask, each of which has different material characteristics: the outer layer is made of spun bond polypropylene, the middle layer is made of Melt-blown polypropylene, and the inner layer is made of a mixture of polypropylene with polyester or cotton will have a different degradation process description with the same treatment. The photooxidation treatment of disposable masks causes significant cracks on their surface, leading to brittleness of the material. These cracks result from the breakdown of polymer chains reacting with hydroxyl radicals (Liu et al., 2022). These cracks can reduce the elasticity of the mask fibers, causing the photo oxidized mask fibers to look more tenuous than those without oxidation (Tsubone et al., 2019). The photooxidation treatment also accelerated the degradation of the masks in soil compared to the masks that were not subjected to similar treatment. This can be observed in Fig. 1 (pots 1, 2, and 3), which show more tenuous mask fibers and more biofilm build-up compared to pots 4 and 5 that were not photo oxidized.\u003c/p\u003e\n \u003cp\u003eMore biofilm formation on oxidized mask surfaces occurs because photooxidation accelerates the biodegradation process. Photooxidation causes mechanical fragmentation of the mask material, producing small fragments that provide more surface for soil microbes to break down the mask polymer (Rizzarelli et al., 2021). In addition, the addition of \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e bacteria and earthworms led to increased biofilm production. \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e bacteria can trigger the rate of bacterial activity in the soil. It can be seen in pots 3 and 4 that the addition of biofilm increased significantly which was marked by the presence of more and more yellow color in the 2nd layer. \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e bacteria are known to be able to degrade various organic and inorganic compounds, and produce enzymes such as proteases and lipases that play a role in breaking down the molecular structure of mask polymers, thus supporting the degradation process (Mohanan et al., 2020).\u003c/p\u003e\n \u003cp\u003eThe addition of earthworms in the soil system, as shown in pot 3 and pot 4, plays a role in accelerating the biodegradation process carried out by bacteria. Earthworms break down organic matter into smaller particles through mechanical digestion (Wang et al., 2021), such digestion results in nutrient-rich excretions often referred to as vermicast. Vermicast contains organic substances and beneficial microbes that can stimulate the formation of bacteria, favorable bacterial growth resulting in a significant increase in biofilm compared to pot 5 and pot 2. In addition, the burrowing activity performed by earthworms also increases soil aeration and moisture, creating more conducive conditions for decomposing microorganisms (Ahmed and Al-Mutairi, 2022).\u003c/p\u003e\n \u003cp\u003eBased on Table 1, the change in thickness of each layer of the disposable mask is seen to decrease significantly due to the photooxidation treatment compared to the control. The photooxidation process involves a reaction between the polypropylene in the mask and oxygen in the air, triggered by exposure to ultraviolet (UV) light from the sun. UV radiation breaks the polymer chains present in polypropylene, generating free radicals, this is a precursor to the degradation of polymer molecules into smaller molecular fragments, such as hydroperoxides and carbonyls. These fragments cause brittleness of the material, so the mask layer begins to crack (Tang et al., 2019). Biodegradation usually consists of three distinct steps: 1) formation of biofilm formation on the plastic surface; 2) decomposition of the plastic into smaller molecules through the action of extracellular enzymes secreted by the microorganisms; and 3) consumption and further metabolism of these smaller molecules these smaller molecules within the cell (Andrady et al., 2022a).\u003c/p\u003e\n \u003cp\u003eThe thickness test was conducted to see the change in thickness of each mask layer after the degradation process. Ultraviolet radiation in the photooxidation process on disposable mask polymers results in damage to the polymer chain which has an impact on the deterioration of plastic mechanical properties including the change of thickness (Wang et al.,\u0026nbsp;2021) .\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 1.\u0026nbsp;\u003c/strong\u003eThickness analysis of disposable mask. Data are shown as mean \u0026plusmn; SD (n = 2).\u003c/p\u003e\n \u003cp\u003e\u003cimg src=\"https://myfiles.space/user_files/69519_bce2c0439cd956a6/69519_custom_files/img1764579852.png\" style=\"max-width: 100%; cursor: pointer; color: rgb(0, 0, 0); font-family: \u0026quot;Times New Roman\u0026quot;; font-size: medium; font-style: normal; font-variant-ligatures: normal; font-variant-caps: normal; font-weight: 400; letter-spacing: normal; orphans: 2; text-align: start; text-indent: 0px; text-transform: none; widows: 2; word-spacing: 0px; -webkit-text-stroke-width: 0px; white-space: normal; background-color: rgb(255, 255, 255); text-decoration-thickness: initial; text-decoration-style: initial; text-decoration-color: initial;\"\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003eIn \u003cstrong\u003etable.1\u003c/strong\u003e. shows a significant decrease in thickness in all mask layers after photooxidation treatment. The decrease also occurred after the disposable masks were buried in the ground, both with the addition of bacteria and without the addition of bacteria. Meanwhile, the masks with photooxidation treatment and continued burial in soil with the addition of bacteria (P2 and P3) showed a greater decrease in thickness when compared to disposable masks without photooxidation both with burial in soil using bacteria and without bacteria. However, photodegradation is an important process in plastic degradation that causes changes in plastic thickness and structural integrity (Sivasankar and Sunitha, 2024).\u003c/p\u003e\n \u003cp\u003eAfter the photooxidation lasted longer, the cracks on the surface of the mask began to deepen and spread to the inside of the mask, reducing the thickness of the mask gradually (Table 2). Macroscopically, the thickness of the mask will decrease as the material structure is destroyed. In addition, the addition of bacteria and earthworms to the biodegradation process in the soil also causes a decrease in thickness in each layer. Bacteria release extracellular enzymes that break down the polymer chains in the mask. The broken polymer chains cause the mask to lose thickness (Mohanan et al., 2020). Meanwhile, the addition of earthworms plays a role in accelerating the biodegradation process by providing a physical and chemical environment where decomposing microbes thrive such as \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e bacteria (Ahmed and Al-Mutairi, 2022). Seen in P4, the addition of bacteria and earthworms resulted in a decrease in thickness compared to P5, without the addition of bacteria and earthworms.\u003c/p\u003e\n \u003cdiv\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 2\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eThickness analysis of disposable mask. Data are shown as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;2).\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eLayer\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"7\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eUntreated Mask\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePhoto-oxidized Mask\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP1 Mask\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP2 Mask\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP3 Mask\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP4 Mask\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP5 Mask\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\u003e1st Layer\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.102\u0026thinsp;\u0026plusmn;\u0026thinsp;0.010\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.077\u0026thinsp;\u0026plusmn;\u0026thinsp;0.007\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.070\u0026thinsp;\u0026plusmn;\u0026thinsp;0.009\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.069\u0026thinsp;\u0026plusmn;\u0026thinsp;0.006\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.070\u0026thinsp;\u0026plusmn;\u0026thinsp;0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.081\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.087\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2nd Layer\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.108\u0026thinsp;\u0026plusmn;\u0026thinsp;0.010\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.087\u0026thinsp;\u0026plusmn;\u0026thinsp;0.006\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.079\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.081\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.079\u0026thinsp;\u0026plusmn;\u0026thinsp;0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.089\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.090\u0026thinsp;\u0026plusmn;\u0026thinsp;0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3rd Layer\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.086\u0026thinsp;\u0026plusmn;\u0026thinsp;0.007\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.067\u0026thinsp;\u0026plusmn;\u0026thinsp;0.008\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.061\u0026thinsp;\u0026plusmn;\u0026thinsp;0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.065\u0026thinsp;\u0026plusmn;\u0026thinsp;0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.061\u0026thinsp;\u0026plusmn;\u0026thinsp;0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.067\u0026thinsp;\u0026plusmn;\u0026thinsp;0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.069\u0026thinsp;\u0026plusmn;\u0026thinsp;0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\"\u003e\n \u003ch2\u003e3.2 FTIR analysis of photo-oxidized and biodegraded mask\u003c/h2\u003e\n \u003cp\u003eOxidative photodegradation (photo-oxidation) of polypropylene is an important stage before reaching the biodegradation stage, due to the fact that biodegradation process in plastics occurs when carbonyl groups have been formed on polymers (Andrady et al., 2022a). In the research of Khoironi et al., (2020), the photo-oxo degradation process is able to provide significant changes in the molecular structure of organic polypropylene with the formation of new groups that are very important in the attack of microorganisms. The formation of carbonyl groups such as carboxylic acids, aldehydes and ketones will increase the moisture (hydrophilicity) of polypropylene, increase amorphous properties, form pores on the plastic surface due to holes and cracks (Esmaeili et al., 2013). Of course, these conditions will facilitate the attack of microorganisms that will work in biodegradation (Mohanan et al., 2020). Photo-oxidation of polypropylene disposable masks produces carbonyl, chain unsaturation and carboxylic acid products, which will initiate the next process where weathering due to this attack will increase the ability to absorb ultraviolet light photo-initiation (Andrady et al., 2022b).\u003c/p\u003e\n \u003cp\u003eAs a consequence of the photo-oxidation process, there is a great influence on the physical and mechanical properties of polypropylene in disposable masks. In the oxygen diffusion controlled photo-oxidation process, crystallinity will also increase with the oxidation rate. The dismemberment of macromolecules involves the formation of smaller molecules that allow easier crystallization. Furthermore, this change will lead to a decrease in molar mass caused by chain cutting due to UV exposure for a long time, thus changing from ductile to brittle behavior which results in easy attack by microorganisms in the biodegradation process (Yousif and Haddad, 2013).\u003c/p\u003e\n \u003cp\u003eFrom Fig. 2. Shows changes in all layers of disposable masks. These changes regarding the formation of new functional groups indicate the uptake of oxygen due to surface vulnerability as a result of ultra-violet radiation. In the first layer of the mask after being treated with photo-oxidation, there are several peaks formed at waves that are not present in the control mask, each of which is 899 cm-3 which is aimed at ketones, then 1102 cm\u003csup\u003e-1\u003c/sup\u003e and 1256 cm\u003csup\u003e-3\u003c/sup\u003e which both point to carboxylic acids, then 1302 cm\u003csup\u003e-3\u003c/sup\u003e which points to esters and 2838 cm\u003csup\u003e-3\u003c/sup\u003e at the -OH group. Similar to what happened in the first layer, some new peaks also appeared in the second layer of disposable masks after photo-oxidation treatment. The appearance of new peaks is indicated by peaks at a wavelength of 899 cm\u003csup\u003e-3\u003c/sup\u003e which is aimed at ketones then 1102 cm\u003csup\u003e-3\u003c/sup\u003e and 1256 cm\u003csup\u003e-3\u003c/sup\u003e which both lead to carboxylic acids and 1302 cm-3 which leads to esters, this also appears in the third layer except 1302 cm\u003csup\u003e-3\u003c/sup\u003e which is not visible in the third layer of disposable masks after photo-oxidation treatment\u003c/p\u003e\n \u003cp\u003eAccording to Zhu et al., (2020), cycles in photo-oxidation will facilitate the attack of microorganisms to produce rapid fragmentation which will lead to complete mineralization in the biodegradation process. The presence of worms and \u003cem\u003epseudomona\u003c/em\u003es bacteria in this study did not appear to have a significant impact after the disposable masks were buried in the soil for a long time. In 2nd layer disposable masks (Fig. 2B) with photo-oxidation treatment (P2, P3), there is a peak at 1302 cm-3 that leads to esters and a peak at 3393 cm-3 that leads to ketones, these two peaks have the same intensity even though the two reactors (P2 and P3) were treated with different organism interventions, it appears that the presence of \u003cem\u003epseudomonas\u003c/em\u003e and worms does not have an impact on the presence of worms. on the other hand, a peak appears to appear in the FTIR analysis results for masks without photo-oxidation (P4 and P5) at a wavelength of 1302 cm\u003csup\u003e-3\u003c/sup\u003e which is directed at the ketone functional group.\u003c/p\u003e\n \u003cp\u003eIn the first layer of disposable masks new peaks appeared only on the masks in reactors P2, P3 and P4 where the reactors received organism treatment, although it seems that organisms do not really play a role in this degradation process but the appearance of peaks at important wavelengths produced by the masks in the reactor indicates the role of the organism. The peaks that appear are at a wavelength of 1164 cm\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e which leads to carboxylic acids and a wave of 1654 cm\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e for P2 which leads to the formation of ketones and a wavelength of 1647 cm\u003csup\u003e\u0026minus;\u0026thinsp;3\u003c/sup\u003e in the three reactors which leads to carbonyl groups.\u003c/p\u003e\n \u003cp\u003eIn the third layer of disposable masks, both masks that have undergone photo-oxidation and those without photo-oxidation treatment do not seem to provide significant results after being buried in the ground. There are no significant peaks that indicate changes in functional groups due to biological or abiotic attacks in the soil, except for the appearance of one peak at a wavelength of 3695 cm-\u003csup\u003e3\u003c/sup\u003e which leads to hydroxyl and carbonyl groups that appear on the mask at P2, P3 and P4.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\"\u003e\n \u003ch2\u003e3.3 Morphological analysis using scanning electron microscopy\u003c/h2\u003e\n \u003cp\u003eTo evaluate the impact of photooxidation on biodegradation efficacy, masks were buried for 100 days, including both treated and untreated masks. Figure 3. illustrates alterations in the morphology of the mask fibers both untreated and photo- oxidized masks post-burial. The changes encompass the formation of biofilms, the emergence of cracks on the mask fibers\u0026apos; surface, and an increase in mask porosity. This may arise from microbial activity, chemical reactions, and physical degradation. Polypropylene mask fibers can serve as a substrate for biofilm development due to their properties. Hydrocarbon-degrading bacteria attaching to the mask\u0026apos;s surface generate enzymes that cleave the long polymer chains in polypropylene into smaller units, thus altering the physical properties of the mask 2425.\u003c/p\u003e\n \u003cp\u003eFigure 3. shows uneven surfaces, cracks and loosen fibers in photo-oxidized masks, which were not appeared in non-photo-oxidized masks. This results align with a study by 6, which reported that disposable masks began to degrade after being exposed to UV irradiation for 36 hour. In current study, the photooxidation process was conducted for 72 hours and assisted by an oxidizing agent (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e). When H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e is exposed to UV light, it breaks down into two hydroxyl radicals (\u0026bull;OH) .These radicals attack the polymer chains of the mask, which over time, can cause the mask to become brittle and loosen its fibers 6\u003c/p\u003e\n \u003cp\u003eThe fine cracks formed through photooxidation cause the mask fiber\u0026apos;s surface rougher. The rough texture of the mask fiber increases the surface area for bacterial adhesion, while the irregularities on the rough surface shield the bacteria, safeguarding them from being displaced by physical disruptions such as water flow from irrigation (Mu et al., 2023). Research indicates that bacteria, including \u003cem\u003eEscherichia coli, Pseudomonas aeruginosa\u003c/em\u003e, and \u003cem\u003eStaphylococcus aureus\u003c/em\u003e, adhere and move more effectively to rough polypropylene fibers compared to smooth ones. The increased movement and adherence of bacteria result in the formation of more biofilms (Cai et al., 2019). In this investigation, the biofilm developed in P3 exhibited no significant difference when compared to P5. This may be affected by soil conditions that facilitate the enzymatic activities of \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e on masks, regardless of photooxidation treatment on masks. Furthermore, in the control mask, the disparity in surface roughness of the mask fibers with and without photooxidation was minimal, resulting in comparable bacterial activity on both treatment. Consequently, additional research is required to determine the optimal duration for the photooxidation process of disposable masks.\u003c/p\u003e\n \u003cp\u003eThe incorporation of \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e into the growth medium augmented biofilm production, as illustrated in Fig. 3, where P2, P3, and P4 exhibit more biofilm accumulation compared to reactors P5 and P1. The introduction of \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e serves as an inoculant, stimulating the rate of bacterial activity in the soil. The incorporation of worms into the growth media enhances the quantity of biofilm produced, as worms optimize the physical and chemical conditions conducive to microbial proliferation. Worms make castings (excrement) that are abundant in organic nutrients and create an optimal environment for bacterial proliferation. These castings possess an increased surface area and comprise water, nutrients, and microbes, which enhance microbial activity. Furthermore, earthworms enhance soil aeration and water retention via their burrows, fostering an environment with elevated oxygen and water availability, both crucial for bacterial metabolism (Buivydaitė et al., 2023; Duan et al., 2024; Zhang et al., 2025).\u003c/p\u003e\n \u003cp\u003eDisposable surgical masks commonly comprise three layers, each constructed from distinct materials with certain characteristics that enhance the mask\u0026apos;s overall efficacy. The outer layer consists of spun-bond polypropylene and possesses hydrophobic properties, which reject water and inhibit the ingress of droplets and larger particles into the mask (Res et al., 2018). The second layer consists of melt-blown polypropylene, formed from ultra-fine fibers that generate a dense, electrostatically charged structure. The third layer of disposable surgical masks comprises spun-bond polypropylene blended with soft polyester or cotton (J. Wang et al., 2023). Figure 3. illustrates that biofilm accumulation is more pronounced in the middle layer than in the other layers. This may result from the material composition and structure of the melt-blown polypropylene utilized in this layer. The melt-blown polypropylene layer possesses a thin and porous structure that more effectively captures particles and retains moisture compared to the outer and inner layers. The accumulated moisture fosters an environment that promotes bacterial proliferation. The increased surface area of the intermediate layer offers more areas for bacterial attachment and biofilm formation. In contrast to the hydrophobic outer layer that repels moisture, the inner layer\u0026apos;s composition enables it to retain moisture and organic matter, including soil nutrients, so facilitating bacterial colonization. The confined particles and moisture create an environment and nutrient source for bacteria, hence improving biofilm formation. The high surface area, moisture retention, and porous characteristics of the middle layer render it more prone to bacterial colonization and biofilm formation than the other mask layers. This aligns with study (Jeong et al., 2021), which shown that bacterial activity in the middle layer exceeded that of the outer and inner layers.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eFrom the data obtained in this study, there is a tendency to have the same phenomenon, where photo-oxidation treatment on disposable mask samples does not provide significant changes after the mask is buried in the ground. Mechanical changes using tests using a microscope, thickness and SEM show the influence of pretreatment on masks that experience photo-oxidation. Significantly, the appearance of biofilm is accompanied by the formation of holes and an increase in pores on the surface of the mask which plays a role in mask fragility. Biological attack also causes the mask fibers to be more tenuous, this shows the increased hydrophilic properties of the mask, it seems that the moisture generated by the biofilm plays a big role in this. The mechanical changes that occur are of course supported by changes in the molecular structure of the mask, where the FTIR test results show the emergence of important functional groups that play a role in the degradation process, namely carboxylates, ketones and esters in Reactor R3, R4 and R5. To find out more precisely about the involvement of organisms in achieving complete biodegradation results, it is necessary to carry out a longer burial process by monitoring the life of the organisms involved and the stability of temperature and humidity.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eConflict of Interest\u003c/h2\u003e\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003ch2\u003eEthical Declaration\u003c/h2\u003e\u003cp\u003eThis study was conducted in accordance with the applicable institutional and national guidelines. The photo-oxidation procedure adhered to ASTM D5208 (Standard Practice for Fluorescent Ultraviolet Exposure of Photodegradable Plastics). Plant physiological responses were assessed using standard, widely accepted physiological evaluation methods, as no specific guideline exists for this type of plant assay. Ethical approval was not required for this research because it did not involve human participants, personal data, or vertebrate animals.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003ch2\u003eConsent to Participate\u003c/h2\u003e\u003cp\u003eNot applicable. No human participants were involved in this study.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConsent to Publish\u003c/strong\u003e\u003cp\u003eNot applicable. This manuscript does not contain any individual person\u0026rsquo;s data in any form (including images, videos, or identifiable information).\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFundings\u003c/h2\u003e\u003cp\u003eThis research was financially supported by The World Class University Research (RWCU) by Diponegoro University, Grant-No: 222\u0026ndash;771/UN7.D2/PP/IV/2025 and KAKENHI (24K00992).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eConceptualization: Hadiyanto, Adian Khoironi; Methodology: Hadiyanto, Adian Khoironi, Inggar Dianratri; Investigation and Data Curation: Falvocha Alifsmara Joelyna, Rifqi Ahmad Baihaqi, Wahyu Zuli Pratiwi, Wahyu Diski Pratama; Formal Analysis: Falvocha Alifsmara Joelyna; Validation: Widayat, Marcelinus Christwardana, Adhelia Intan Sabhira; Resources and Supervision: Hadiyanto, Adian Khoironi, Widayat; Writing Original Draft: Adian Khoironi; Writing Review \u0026amp; Editing: Adian Khoironi, Marcelinus Christwardana, Hadiyanto, Tomoya Kataoka; Project Administration and Funding Acquisition: Hadiyanto, Adian Khoironi\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThe authors would like to express their profound gratitude to the laboratory assistants at the Center of Biomass and Renewable Energy Laboratory for their essential technical support for the entirety of this research endeavor. Their proficiency and assistance have been vital in the accomplished implementation of this project.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe data supporting the findings of this study are available from the corresponding author upon reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAhmed N, Al-Mutairi KA. Earthworms Effect on Microbial Population and Soil Fertility as Well as Their Interaction with Agriculture Practices. Sustain. 2022;14:1\u0026ndash;17. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/su14137803\u003c/span\u003e\u003cspan address=\"10.3390/su14137803\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAkber Abbasi S, Khalil AB, Arslan M. 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J Hazard Mater. 2020;383:121065. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jhazmat.2019.121065\u003c/span\u003e\u003cspan address=\"10.1016/j.jhazmat.2019.121065\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\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":"biodegradation, disposable mask, photo-oxidative degradation, soil system","lastPublishedDoi":"10.21203/rs.3.rs-8082479/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8082479/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe increase in the amount of disposable mask waste causes problems in reducing soil quality. In the terrestrial environment, disposable masks will experience various stages of the degradation process which are influenced by abiotic factors involving sunlight and air as well as biotic factors involving organisms in the soil. There are not many studies that reveal the success of the degradation of disposable masks in nature, so this research was conducted to look at the influence of natural factors in the degradation process of disposable masks. The experiment, which was carried out for 100 days, involved \u003cem\u003epseudomonas bacteria\u003c/em\u003e, worms and plants to condition the degradation media to resemble natural conditions. Changes in disposable masks due to chemical and biological degradation are shown from the results of use tests using microscope, thickness, FTIR and SEM. The results of the analysis showed that there was significant damage to disposable masks through examination with a microscope during the treatment of the masks using a photo oxidation process followed by treatment involving worms and bacteria. Conversely, the formation of important ketone, ester and hydroxide groups after the photooxidation process as shown by the FTIR results have not significant gap with the result in mask without photo-oxidation treatment. Furthermore, \u003cem\u003ePseudomonas\u003c/em\u003e bacteria were able to reduce the thickness of disposable masks effectively compared to single treatment using worms. This research shows the very important role of oxygen availability and sunlight in the degradation process of disposable masks before they reach the complete stage of degradation.\u003c/p\u003e","manuscriptTitle":"Photo-oxidative degradation of disposable mask and the effect on the biodegradation process in soil system","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-12-01 09:07:15","doi":"10.21203/rs.3.rs-8082479/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":"eec76995-fd62-4e65-87a6-4bd0151a54ad","owner":[],"postedDate":"December 1st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-04-01T08:11:43+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-01 09:07:15","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8082479","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8082479","identity":"rs-8082479","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}