Comparative analysis of antimicrobial properties of maggot (Musca domestica) crude extract, maggot chitosan and chitosan nanoparticles

Comparative analysis of antimicrobial properties of maggot (Musca domestica) crude extract, maggot chitosan and chitosan nanoparticles | 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 Comparative analysis of antimicrobial properties of maggot (Musca domestica) crude extract, maggot chitosan and chitosan nanoparticles Maira Munir, Saffora Riaz, Saima Samra, Ammara Zeb This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6084200/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 Background: Maggot metabolites exhibit strong antibacterial and pro-inflammatory properties, making them a significant focus of scientific research. Aim: This study aimed to compare the antimicrobial properties of maggot extracts (MEx), maggot chitosan (MCs), and maggot chitosan nanoparticles (CNPs). Methodology: Maggot chitosan with 90.57% degree of deacetylation was extracted. CNPs were synthesized via ionotropic gelation with sodium tripolyphosphate and the characterization results confirmed their semi-crystalline structure with particle size of 262 nm. Results: GC-MS analysis of MEx identified twenty bioactive compounds in MEx. By using disc diffusion method, all treatments showed significant antibacterial activity ( P = 0.0004 ) against different bacterial strains. MEx exhibited the highest antibacterial activity against E. coli (2021), P. aeruginosa (101 and 310), S. aureus (723) and K. pneumoniae (310) with a zones of inhibition 20.3 ± 0.3 mm, 29.6 ± 0.6 mm, 18.6 ± 0.3 mm, 18.6 ± 0.3 mm and 22.6 ± 0.3 mm respectively. MCs showed the highest antibacterial activity against E. coli strains (1876 and 1609) with zones of inhibition 31.6 ± 0.6 mm and 20.3 ± 0 mm. The antibacterial activity of CNPs was comparatively lower than that of MEx and MCs. In antifungal susceptibility tests, all treatments were significantly sensitive to both Aspergillus molds ( P = 0.0001 ). However, MEx showed the highest antifungal activity against A. flavus and A. niger with zones of inhibition 23.3 ± 0.3 mm and 23 ± 0.5 mm. Findings: The findings suggest that these cost-effective medications would be very effective in preventing infections in the body. Additionally, they have minimal side effects. Further research and development could lead to new, effective medications based on these agents. Biological sciences/Biochemistry Biological sciences/Microbiology Biological sciences/Zoology Health sciences/Diseases Physical sciences/Nanoscience and technology alternative therapeutics biomedical applications pathogen inhibition bioactive compounds natural antimicrobial agents 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 Figure 16 Figure 17 Figure 18 Figure 19 Figure 20 Figure 21 Figure 22 Figure 23 Figure 24 Figure 25 Figure 26 INTRODUCTION Maggots are the larval stage of the Musca domestica species that belong to the Diptera: Muscidea family. The common housefly is acknowledged for its role as a sanitation pest. However, because of the short life span, high potential for reproduction, and efficient breakdown of organic waste, it has also been emerged as a valuable source of protein, chitin, and chitosan [ 1 – 2 ]. Housefly larvae have been a source for the extraction of valuable materials like peptides, chitosan, phospholipids, and antibiotics. Based on initial observations, it was determined that roughly 55% of dried housefly larvae's composition is protein, 9.1% is crude chitin, and 8.8% is fat [ 3 – 4 ]. Housefly larvae are well-known for their non-toxic properties. They have been clinically used to treat various conditions, such as ecthyma, wounds, and bacterial infections [ 5 – 6 ]. Due to the ease of access and low cost associated with breeding Musca domestica , it has been proposed as a potential alternative to Lucilia sericata for this therapy [ 7 ]. Maggot therapy, widely known for its powerful ability to fight off different infections and emerges as the fastest and most effective method for clearing necrotic tissue and treating infections [ 8 ]. Maggots have been observed to reduce the amount of bacteria in wounds due to their multiple antibacterial actions [ 9 ]. Maggot excretions can terminate several pro-inflammatory reactions by neutrophils, including degranulation, chemotaxis, respiratory burst, and integrin expression [ 10 ]. Chitinous materials such as chitin and chitosan are among the planet's second-most abundant biopolymers, following cellulose. Their versatility spans over two hundred applications, ranging from agriculture to biomedicine, cosmetics, food, and textiles, show potential as industrial effluent refiners and chelating agents [ 11 – 12 ]. Chitin mainly comes from the shells of crustaceans such as shrimp and crabs [ 13 ]. However, these shells have a lot of calcium, wax, and pigment, which makes it expensive to extract chitosan from them. Comparatively, housefly larvae make chitin extraction easier when contrasted with shrimp or crabs [ 14 ], as they have reduced amounts of ash, crude fat, and crude protein [ 15 ]. Chitosan is obtained from the process of deacetylation of chitin, a process that removes acetate groups. The shells of housefly larvae are rich in chitin, making them a significant source of chitosan [ 16 ]. The depolymerization of chitosan has significant interest as it enhances the water solubility of the resulting products. Furthermore, numerous chemical alterations have been investigated to enhance the solubility of chitosan and broaden its potential applications [ 17 – 18 ]. Chitosan and its oligosaccharides offer a lot of benefits, such as fighting tumors, protecting the nervous system, combating fungal infections, and reducing inflammation [ 19 ]. In recent years, chitin, chitosan, and their derivatives have gained significant attention for their antimicrobial properties against diverse microorganisms, including bacteria, yeast, and fungi [ 20 ]. Chitosan shows its ability to fight bacteria best in acidic conditions because it doesn't dissolve well above pH 6.5 [ 21 ]. It is more effective and has increased efficacy in combating gram-positive bacteria in comparison to gram-negative ones [ 22 ]. The antibacterial and antifungal profile of chitosan and its derivatives have made them significant components in commercial disinfectants [ 23 ]. Chitosan has abundant advantages over other disinfectants because it is highly effective against bacteria and less harmful to mammalian cells [ 24 ]. The antimicrobial activity of chitosan is the most significant bioactivity and it has been used in the development of biomedical materials as well as in improving the functionality of other polymeric materials such as fibers and food preservation [ 25 – 26 ]. Chitin and chitosan, both, have the ability to stimulate the host's defense system and protect the host against pathogen invasion [ 27 ]. Two main mechanisms have been proposed for how chitosan inhibits microbial cells. One is a positive charge present on chitosan that disrupts bacterial metabolism by sticking to the surface of bacterial cells [ 28 – 29 ]. Another reason is that chitosan adsorbs, preventing the transcription of RNA from DNA by binding to DNA molecules [ 19 ]. Chitosan is an ideal material for enzyme immobilization because of its enhanced resistance to chemical degradation and the ability to prevent metal ions from interfering with enzymes [ 30 ]. The amino functional group found in chitosan is suitable for immobilizing enzymes [ 31 ]. Research on the pro- and anti-inflammatory properties of chitosan and its derivatives revealed that they suppress the development of colitis and increase the induction of the anti-inflammatory IL-10 cytokine in animal blood [ 32 ]. Nanotechnology has gained prominence in several fields because of its unique qualities and the growing problem of microbial drug resistance. Nanoparticles (NPs) can interact with biological molecules and, most importantly, microorganisms due to their unique physical and chemical properties that allow them to pass through barriers [ 33 ]. Chitosan nanoparticles (CNPs) combine the qualities of nanoparticles with those of chitosan, including surface and interface effects, small size, and quantum size effects [ 34 ]. The surface-to-volume ratio of chitosan is higher at the nanoscale, which increases its affinity for bacteria and fungi. Chitosan nanoparticles directly interfere with microbial cell membranes, cell walls, key proteins, and enzymes, capable of inhibiting pathogen growth and inducing cell death through mechanisms distinct from conventional antibiotics, which may have developed resistance. The size, shape, and chemical properties of nanoparticles can be manipulated to enhance these molecular interactions, optimizing their overall effectiveness [ 33 , 35 ]. Consequently, a variety of medications can be produced using nanoparticle manufacturing techniques [ 36 ]. Chitosan-based nanoparticles show potential in medicine and the food industry because materials that are nanoscale in size acquire unique characteristics that set them apart from bulk materials and isolated atoms [ 37 ], and are employed in the control of ocular infections, gastrointestinal disorders, lung disorders, cancer, and drug delivery to the brain [ 38 – 39 ]. When chitosan comes into contact with anions, it has the ability to form gel and beads. Due to this attribute, it can be used to deliver drugs [ 40 ]. The development of a wide range of colloidal delivery vehicles has been made possible by the potential use of CNPs as carriers [ 41 ]. In biological media, chitosan nanoparticles (CNPs) can withstand biological barriers and prevent macromolecules from degradation. It can also deliver medicinal products or macromolecules to a target location by controlled release [ 42 ]. Small-sized CNPs also contribute to its efficiency in interfacial interaction with cell membranes because the tiny particles are taken up by the cell through endocytosis [ 43 ]. CNPs can increase the bioavailability of pharmaceutical products by altering the pharmacokinetics and insulating the encapsulated drugs [ 44 ]. In a dose-dependent manner, chitosan nanoparticles can effectively cause human keratinocytes and monocytes to produce cytokines [ 45 ]. CNPs are an excellent environmentally friendly material with non-toxic bioactive and natural materials with exceptional physicochemical, antimicrobial, and biological qualities [ 41 ]. CNPs have been studied extensively in the fields of biomedical materials and tissue engineering because of their good water solubility, biocompatibility, biodegradability, antimicrobial, film forming properties, hemostasis, wound healing, and anti-inflammatory effects. They were successfully used as a drug delivery system. Target therapy may benefit from the use of these NPs [ 46 ]. Recent studies underscore the potential of maggot therapy and chitosan-based treatments for different infection control. The efficacy of maggot extract (MEx) and chitosan in enhancing wound healing and exhibiting antimicrobial properties [ 47 – 48 ]. Maggot chitosan (MCs) and chitosan nanoparticles (CNPs) are effective against various bacterial strains [ 47 ]. Additionally, The potential of chitosan nanoparticles in biomedical applications, which is supported by your UV analysis showing specific absorbance characteristics for CNPs [ 48 ]. The study aimed to compare the antimicrobial properties of maggot extract, maggot chitosan, and chitosan-based nanoparticles. The objectives were: to extract chitin from Musca domestica for chitosan preparation, to synthesize and characterize chitosan nanoparticles using UV-visible spectroscopy, XRD, FTIR, and SEM, and to evaluate and compare the antimicrobial effectiveness of maggot extract and chitosan nanoparticles against E. coli , Pseudomonas aeruginosa , Staphylococcus aureus , and Klebsiella pneumoniae . MATERIALS AND METHODS The experiment was conducted in the Entomology Lab of the Department of Zoology at Lahore College for Women University, Lahore. Maggot collection A housefly breeding site was established in a plastic jar. Flies were allowed to feed on chicken offal. The breeding sites were left undisturbed for a week, allowing the flies to lay eggs, hatch, and develop into larvae or maggots. During this period, the breeding site was regularly monitored to check the optimal environmental factors, such as temperature and moisture levels, to ensure ideal conditions for maggot growth. After 7 to 8 days, the breeding sites were carefully examined, and the mature maggots, second and third larval instars were carefully collected. After the collection, maggots were killed by placing them in hot water at 80°C for 3 to 5 minutes and then washed with saline water to remove any debris. The maggots were dried in an air-dried oven at 55°C. Once dried, the maggots were stored in airtight containers or sealed bags at room temperature for future use. Preparation of crude extract of maggots The crude extract of maggots was prepared following the methodology of [6], with some modifications. About two hundred grams of maggots were selected and crushed in a mortar and pestle. The crushed maggots were added to a conical flask and two volumes of 10X Tris NaCl EDTA (TNE) buffer (pH 7.5-8) precooled at 4 o C were added. The mixture was homogenized for 15 minutes at room temperature. After sufficient homogenization, the conical flask was covered with aluminum foil and it was left at room temperature for 24 hours. The next day, the homogenate was transferred to a sterile falcon tube and centrifuged in a refrigerated centrifugal machine (MSE Harrier 18/80) at 6000 rpm for 20 minutes at 4 o C. The supernatant was transferred to a new falcon tube and again three volumes of precooled 10X TNE buffer were added. This mixture was placed in a hot water bath at 70 o C for 10 minutes. After this, the mixture was cooled and again centrifuged at 6000 rpm for 20 minutes at 4 o C. The resulting supernatant, which was the pure crude extract of maggots, was transferred to a new sterile falcon tube. The extract was frozen for 24 hours in a freezer, then freeze dried at -80 o C. The frozen dried powder was stored at -20 o C. GC-MS analysis of crude extract GC-MS analysis of maggot extract was performed using a SHIMADZU GCMS QP-2010 system. An analytical column HP 5MS (30m x 0.25mm x 0.25mm) was employed, with the GC equipped with a manual splitless injector set at 240°C. The injector volume was 2 µl, and the pressure was maintained at 11.567 psi, with a split ratio of 200:1. The gas saver was turned off, and the total flow rate was set at 204 ml/min, with helium used as the carrier gas. For the oven program, the temperature was initially set at 40°C with a hold time of 1 minute, followed by ramping to 280°C at a rate of 10°C/min and holding for 5 minutes at this temperature. The mass spectrometer (MS) was set to a range of m/z 35-450, with a solvent delay of 0.1 minutes. The MS quad temperature was maintained at 150°C. The GC inlet temperature was set to 280°C for optimal performance. The results was observed accordingly. Isolation of chitin from maggots Chitin was isolated from the larva (maggots) of house flies [4], and this process was used with some modifications and involved several steps. Defatting One hundred grams of dried maggots were collected and crushed into a fine powder using a mortar and pestle. The dried, crushed maggots were transferred to a beaker. About 500 ml (v/v) of the chloroform (Sigma-Aldrich CAS No. 67-66-3) and methanol (Sigma-Aldrich CAS No. 67-56-1) mixture (7:3) was added to the crushed, dried maggots. This mixture was placed on a hot magnet plate and stirred for about 4 hours at 30 o C. After 4 hours, the mixture was allowed to cool down to room temperature and filtrated. The filtrate was washed with distilled water until a neutral pH was attained. Air-dried the defatted crude chitin in an oven at 45 o C for 24 hours and measured the dry weight of the obtained material. Deproteinization The dried, obtained material was placed in a 5% (w/w) NaOH (Sigma-Aldrich Lot# STBH4233) solution in a ratio of 1:10. This mixture was stirred for about 2 hours at 75 o C. After 2 hours, the mixture was allowed to cool down to room temperature, and then the mixture was filtrated and all the dissolved protein content was discarded. The filtrate was washed with distilled water until a neutral pH was attained. The crude chitin was air-dried at 45 o C for 24 hours. The dry weight of the obtained material was measured using a weight balance. Demineralization The obtained crude chitin was placed in 2% (v/v) HCl (Sigma-Aldrich CAS No. 7647-01-0) in a ratio of 1:10. This mixture was placed on a hot magnetic plate and stirred for about 2 hours at 27 o C. After 2 hours, the mixture was cooled to room temperature and then it was filtrated. Filtrate was washed with distilled water until its neutral pH was maintained. The obtained material was air-dried at 45 o C for 24 hours. The weight of the obtained air dried chitin was measured. Preparation of chitosan from isolated chitin Chitosan was prepared from the isolated chitin by the process of deacetylation [49], with some modifications in methodology. Deacetylation of chitin The obtained chitin was placed in a 50% (w/w) NaOH (Sigma-Aldrich Lot# STBH4233) solution. This suspension was stirred so that chitin was completely soaked in the solution and left at room temperature for about 30 minutes. The suspension was placed on a hot magnetic plate and allowed to stir continuously at 100 o C for about 2 hours. After 2 hours, the suspension was subjected to filtration and the filtrate was washed until a neutral pH was maintained. This obtained material, chitosan, was air-dried completely at 45 o C for 12 hours. The obtained chitosan was measured after complete air drying. Decolorization The process of decolorization was performed in order to remove excess pigments from chitosan. The obtained chitosan was placed in a 1% CH 3 COOH (Sigma-Aldrich CAS No. 144-55-8) solution in a ratio of 1:100. This mixture was placed on a hot magnetic plate and allowed to continuously stir for about 30 minutes at 27 o C. The decolorized chitosan was filtrated and washed to maintain a neutral pH. Repeat the process twice for 10 minutes to remove excess pigments. The decolorized chitosan was air dried at 45 o C for 12 hours. The decolorized chitosan was subjected to a ball milling machine, and a fine powder of the prepared chitosan was made at 200 rpm speed for 40-45 minutes. FT-IR analysis of chitosan to determine degree of deacetylation The obtained chitosan was subjected to FT-IR analysis and it was done on IRTracer-100 FTIR Spectrophotometer-Shimadzu, to determine its peaks and degree of acetylation was determined by using the formula given by [50]. Preparation of chitosan nanoparticles Chitosan nanoparticles were prepared using the ion-gelatin method [51], with some modifications. Different concentrations of chitosan (0.1%, 0.2%, 0.3%, 0.4%, and 0.5%) were prepared by dissolving the respective amount of chitosan in 1% (v/v) glacier acetic acid (BDH Laboratory Supplies, Lot# B261651423) at a temperature of 60°C on a hot magnetic plate for 30 minutes at a speed of 900 rpm. After 30 minutes, the chitosan solution was sonicated in an ultrasonic water bath for 30 minutes to remove excess bubbles and ensure a homogeneous solution. The pH of the chitosan solution was adjusted to 5 using a 20% (w/w) NaOH (Sigma-Aldrich Lot# STBH4233) solution. A 0.1% sodium tripolyphosphate (TPP) (Sigma-Aldrich CAS No. 23-85-03) solution was prepared, and its pH was also adjusted to 5 using 0.1M HCl (Sigma-Aldrich CAS No. 7647-01-0). TPP was used as a cross-linker. TPP was gradually added drop by drop into the chitosan solution at room temperature using volume ratio of 1:1 at a speed of 700 rpm on a hot magnet plate. The dripping time should be less than 5 minutes. The solution was stirred on a hot magnetic plate at 700 rpm for 30 minutes. Subsequently, the chitosan-TPP mixture was centrifuged in a refrigerated centrifuge machine (MSE Harrier 18/80) at 6000 rpm for 15 minutes at 4°C. The pellet was collected by discarding the supernatant. Purification of chitosan nanoparticles Deionized water was added to a collected palette, and it was vortexed for a few seconds to dissolve the palette in deionized water. The palette was again refrigerated and centrifuged (MSE Harrier 18/80) at 6000 rpm at 4 o C for 15 minutes. The supernatant was discarded, and the pure palate was collected. The collected palette was the chitosan nanoparticles. Freeze drying of chitosan nanoparticles Glucose (Systerm CAS No. 50-99-7) 5% and sucrose (Sigma-Aldrich CAS No. 57-50-1) 5% were added to chitosan nanoparticles in a volume ratio of 1:1 as cryo-protectants. The nanoparticles were freeze-dried at -80 o C for 24 hours and stored at -20 o C. Characterization of nanoparticles These chitosan nanoparticles were characterized at the Central Research Lab at Lahore College for Women University. Different characterization processes that were used to characterize nanoparticles included the following techniques: X-ray diffraction (XRD) The obtained nanoparticles were subjected to X-ray Diffractometer in order to assess the crystalline structure of the nanoparticles. This technique directed X-rays at the sample and measured the diffraction patterns to determine the arrangement of atoms within the crystal lattice. The resulting data provided information on the crystalinity, phase composition, and other structural parameters of the nanoparticles. Scanning electron microscopy (SEM) SEM was employed to visualize the morphology of the nanoparticles. This method involved scanning the surface of the sample with a focused beam of electrons, which interacted with the atoms in the sample to produce images with high resolution and depth of field. SEM images allowed for the observation of the size, shape, and surface characteristics of the nanoparticles. Fourier-transform infrared spectroscopy (FTIR) FTIR was used for identifying functional groups and the chemical composition of the nanoparticles. In this technique, the sample was exposed to infrared radiation machine (IRTracer-100 FTIR Spectrophotometer-Shimadzu), and the resulting absorption spectra were analyzed to identify specific molecular vibrations and chemical bonds present in the sample. This analysis provided insights into the chemical structure and composition of the chitosan nanoparticles. UV-visible spectroscopy UV-Vis spectroscopy was employed to analyze the optical properties of the nanoparticles. The sample was exposed to ultraviolet and visible light, and the absorption spectra were recorded to determine the electronic transitions and band gaps within the nanoparticles. The results of these analyses were observed to ensure the proper characterization of the chitosan nanoparticles. Assessment of antibacterial activity Bacterial strains Bacterial strains were obtained from Institute of Microbiology, University of Veterinary and Animal Sciences, Lahore. These strains were Escherichia coli (1609, 1874, 2021) Pseudomonas aeruginosa (101, 310) Klebsiella pneumoniae (310) Staphylococcus aureus (723) Sterilization All glassware (including beakers, flasks, test tubes, and glass petri plates), cotton, cotton buds, Eppendorf tubes, Micropipette tips, forceps, and the inoculum loops, forceps were autoclaved at 121 °C and 15 psi for 15 minutes to eliminate all microorganisms and contaminants. Preparation of media Various nutrient media were prepared to provide the necessary nutrients and growth conditions to cultivate bacteria. These media served as the basis for isolating and identifying bacteria. Nutrient agar: Nutrient agar media was prepared by dissolving 14g of weighed nutrient agar (Scharlau Lot# 18155) in 500 ml of distilled water in a conical flask. The solution was thoroughly mixed in order to dissolve it, and after that, it was sterilized in an autoclave at 121 o C for 15 minutes. Nutrient broth: The thirteen grams of nutrient broth (Sigma-Aldrich Lot# 8276350) powder were measured using a weight balance and added to a conical flask. It was then dissolved in 1000 ml of distilled water and gently heated to create a clear solution. The flask was covered with aluminum foil, and the media were autoclaved at 121 °C for 15 minutes. Eosin methylene blue (EMB) agar: EMB agar (Lab M Limited Batch No. 111833/201) was prepared by measuring 37.5 grams of eosin methylene blue agar and added to a conical flask. About 1000 ml of distilled water was added in the conical flask. The suspension was soaked for 10 minutes. After that, it was heated and mixed until a uniform mixture was formed. Sterilization was achieved by autoclaving at 15 psi of pressure (121 °C) for 15 minutes. MacConkey agar: About 51.6g of MacConkey agar (HIMEDIA Lot No. 0000187708) was dissolved in 1000 ml of distilled water and heated with constant stirring to completely dissolve the media. It was then autoclaved at 121°C temperature and 15 psi pressure for 15 minutes. Blood agar: About 40 g of blood agar base (Sigma-Aldrich Lot # 9874589) was measured using a weight balance. The measured blood agar base was added to a sterilized conical flask containing distilled water. The suspension was boiled until it dissolved completely. It was then autoclaved at 121°C for 15 minutes. After autoclaving, the mixture was cooled to 45–50°C, and 6% sterile horse blood was aseptically added. Preparation of media plates The laminar flow cabinet was cleaned and disinfected with ethyl alcohol (Sigma-Aldrich CAS No. 64-17-5). UV light was turned on for 10 to 15 minutes and then switched off. Two burners were lit on both sides of the cabinet, and the blower and fluorescent light were activated. Autoclaved media were placed inside the cabinet and poured into sterilized petri plates, which were covered until they solidified. The petri plates were sealed with paraffin wax or tape to prevent contamination and placed upside down in an incubator at 37 °C for 24 hours. Inoculum development Nutrient broth (3-4ml) was taken into each sterilized test tube with the help of a micropipette. Inoculated the test tubes with a pure culture of test bacterial strains and incubated at 37°C in a shaking incubator for 18- 24 hours. Antibacterial activity of treatments The antibacterial activity of the maggot extract (MEx), maggot chitosan (MCs) and chitosan nanoparticles (CNPs) against different bacterial strains was determined using the disc diffusion method. Preparation of filter paper discs Sterile filter paper was cut into 6mm discs and soaked them in different concentrations of MEx (2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), 10 mg/ml (D)), MCs (2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), 10 mg/ml (D)), CNPs (2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), 10 mg/ml (D)), and saline water for negative control. For positive control, antibiotic susceptibility discs of imipenem (10 mcg) were used. Ciprofloxacin (5 mcg) was used as a resistant group. Disc-diffusion method In this method, a loop full of bacterial culture was taken from stock and inoculated in nutrient broth at 37°C for 24 hours. Put the broth medium on a rotatory shaker overnight at 200 rpm. After 24 hours, swab the broth media on nutrient agar using a cotton swab. Different agar plates for each bacteria were prepared and grouped into different categories: Positive control group : exposed to standard imipenem (10 mcg); Negative control group : exposed to saline water; Resistant groups : exposed to standard Ciprofloxacin (5 mcg); Treatment group I : exposed to different concentrations of maggot extract (MEx); Treatment group II : exposed to different concentrations of maggot chitosan (MCs); Treatment group III : exposed to different concentrations of chitosan nanoparticles (CNPs). Filter paper discs were applied to agar plates corresponding to each group. The discs were gently pressed onto the agar surface to ensure even distribution and incubated at 37 °C for 24 hours. After overnight incubation, zones of inhibition were measured in millimeters and compared with those of control groups. Morphological identification of bacterial strains In the study, bacterial colonies were morphologically identified after 24 to 48 hours of incubation on different agar petri plates. The bacterial colony size, color, and shape were observed to identify the bacterial species. Microscopic identification of bacterial strains Gram staining was used to microscopically identify bacteria into two large groups, gram-positive and gram-negative bacteria. Gram staining To prepare the sample, a drop of saline and a lapful of culture were mixed on a slide to form a smear by gently stirring. The smear was then fixed by passing it through the flame. Subsequently, the smear was flooded with crystal violet and left for 1 minute. Afterward, the slide was tilted slightly and rinsed with distilled water. Next, the slide was flooded gently with grams of iodine and left for 1 minute before being rinsed again with distilled water. At this stage, the smear appeared purplish in color. The slide was then decolorized by drop wise addition of 95% alcohol onto it for 5–10 seconds. After rinsing with distilled water, the slide was gently flooded with the counter-stain safranin for 45 seconds and then rinsed again with distilled water. Finally, the slide was blotted dry with bibulous paper and analyzed under a light microscope at different magnification. Gram-positive bacteria stained purple, while gram-negative bacteria retained safranin and appeared pink in color . Assessment of Anti-fungal Property Collection of fungal strains Fungal strains were obtained from Entomology lab of the department of zoology, Lahore College For Women University, Lahore, Pakistan. The obtained fungal strains were of Aspergillus niger Aspergillus flavus Preparation of media In order to cultivate fungi, SDA media were prepared to provide the necessary nutrients and growth conditions. The SDA media served as the foundation for the isolation and identification of fungi. Preparation of Sabouraud Dextrose Agar (SDA): Measured about 65g of SDA (Sigma-Aldrich CAS No. 57-48-7) and dissolved it in 1000ml of distilled water. The mixture was heated until completely dissolved and then autoclaved at 121°C and 15 psi pressure for 15 minutes. Preparation of petri plates After cleaning the laminar airflow chamber with ethyl alcohol (Sigma-Aldrich CAS No. 64-17-5), the UV light was turned on for 15 minutes. After 15 minutes, it was turned off, and autoclaved media were placed inside the laminar flow. The two Bunsen burners were lit, and then the media were poured into the petri plates, filling up to three-quarters of each plate. The plates were allowed to solidify in the chamber. After solidification, the plates were sealed with paraffin wax, labeled, and left to set in the incubator upside down for 24 hours at 27°C. Antifungal activity of treatments The antifungal effect of maggot crude extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs) were assessed against two different molds fungus. Antifungal activity was assessed using the disc diffusion method. Preparation of filter paper discs Sterilized filter paper was cut into 6 mm round discs and soaked in different concentrations of MEx (2.5 mg/mL (A), 5 mg/mL (B), 7.5 mg/mL (C), 10 mg/mL (D)), MCs (2.5 mg/mL (A), 5 mg/mL (B), 7.5 mg/mL (C), 10 mg/mL (D)), and CNPs (2.5 mg/mL (A), 5 mg/mL (B), 7.5 mg/mL (C), 10 mg/mL (D)). Saline water was used as a negative control and 1 mL of 2% ketoconazole (10 µg/mL) as a positive control. These concentrations were prepared in distilled water. Disc-diffusion method From the previous 7-day-old cultured plates of Aspergillus strains, a small amount of fungal mold was taken with the help of a sterilized swab and swabbed all around the petri plates. After swabbing, the different SDA petri plates were divided into the following groups: Positive control group: Exposed to ketoconazole (10 µg/mL) as a positive control and saline water as a negative control. Treatment group I: Exposed to different concentrations of MEx. Treatment group II: Exposed to different concentrations of MCs. Treatment group III: Exposed to different concentrations of CNPs. Filter paper discs corresponding to each group were applied to the SDA plates. The discs were gently pressed onto the agar surface to ensure even distribution and incubated at 27°C for 5 days. Measurement of growth inhibition zones After the incubation period, the diameter of the fungal colonies on each plate was measured and recorded, and these measurements were compared to those of the control plates. Macroscopic identification of fungi The morphological characters of each colony developed after 6 days. The fungal isolates were identified using standard mycological methods that involve a combination of macroscopic and microscopic examinations. The colony features (color, shape, size, and hyphae) were macroscopically observed in Petri plates to study the fungal morphology. Microscopic examination of fungi The microscopic examination was done under a microscope after staining. A glass slide with 70% alcohol and lacto phenol blue received a fungal specimen via a sterile needle. After being covered with a coverslip, microscopic features were observed under a light microscope. Statistical analysis The data was analyzed using GraphPad Prism 8.0.2 (263), IBM SPSS Statistics 25, Microsoft Excel (2013), and Origin 2018 64 Bit. Means, standard deviations, and standard errors were calculated. Statistical tests, including ANOVA, were employed to assess the antibacterial and antifungal effectiveness of maggot extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs). RESULTS GC-MS analysis of crude extract The qualitative GC-MS analysis of the maggot crude extract revealed 20 distinct peaks; these peaks were identified by comparing their retention times (RT) with standard peaks using MS Interpreter (Ver. 3.4.5.) software. The quantitative analysis of the peaks provided the average concentration of these compounds in the extract solution (MEx). In addition to their antibacterial, antifungal, and anti-inflammatory properties, these compounds exhibit a wide array of bioactive effects suitable for potential medicinal applications. For example, several alcohols and esters found in the extract demonstrate antioxidant potential, which could be beneficial in addressing conditions related to oxidative stress. Furthermore, certain compounds, like cyclic lactones and specific fatty acid derivatives, showed promising neuroprotective properties. Table 1 illustrates all the summarized results of GC-MS of maggot crude extract compounds with their retention time, molecular weight, and formula. Chitosan preparation Chitosan was prepared through a series of different steps, with each step resulting in a significant reduction in material weight. It showed the degree to which each method separated and purified the chitosan from the original maggot material. The yield of each processing step are summarized in Table 2 and Figure 1 detailing the materials obtained along with the corresponding dry weights. Measurement of degree of deacetylation of chitosan The prepared chitosan underwent FT-IR analysis, and the resulting data were utilized to generate a graph spectrum between wavelengths of 4000 cm -1 and 500 cm -1 on the x axis and transmittance on the y axis using Origin 2018 64Bit software, as shown in Figure 2. In the FTIR (Fourier Transform Infrared) analysis of chitosan, several characteristic peaks were observed. Notable peaks included a broad absorption around 3350-3500 cm⁻¹, corresponding to O-H and N-H stretching vibrations, and a peak at 1650 cm⁻¹ associated with the amide I band from C=O stretching vibrations. Additional significant peaks were observed at 1580 cm⁻¹ for N-H bending vibrations, 1420 cm⁻¹ for CH2 bending vibrations, and 1320 cm⁻¹ for C-N stretching vibrations or the amide III band. Peaks at 1150 cm⁻¹ and 1080 cm⁻¹ represented C-O-C and C-O stretching vibrations, respectively, which were indicative of glyosidic linkages in the chitosan backbone. These specific peaks are important in confirming the chemical structure and composition of chitosan, as well as it provide insight into its degree of deacetylation and impurities present in the sample. According to the equations proposed by [50], the degree of deacetylation of the prepared chitosan sample was determined to be 90.57%. Preperation of chitosan nanoparticles The chitosan nanoparticles were prepared using the ion-gelatin method. At concentration of 0.1% and 0.2%, no pellet formation was observed after refrigerated centrifugation. At 0.3% chitosan, a small pellet briefly formed but rapidly disintegrated, preventing its collection for further analysis. However, at a concentration of 0.4%, a stable pellet was successfully observed and collected. The palette at 0.4% chitosan maintained its structure, and this palette was used for further characterization and was used in the further study. At 0.5% chitosan concentration, chitosan was not dissolved properly in the 1% acetic acid solution, and its incomplete dissolution led to the formation of a suspension at the bottom of the flask. Due to this suspension, the cohesive pellet was formed on post-centrifugation, indicating insufficient dissolution in the solution. Characterization results of nanoparticles The characterization results of chitosan nanoparticles are discussed below. FTIR analysis The FTIR spectra of chitosan nanoparticles (CNPs) revealed significant changes induced by interactions with sodium tripolyphosphate (STPP), which were crucial for their structural and functional properties. Key observations included intensified O-H stretching vibrations at 3416 cm -1 , indicating enhanced hydrogen bonding facilitated by STPP. Shifts in NH2 and OH stretching vibrations to 3292 cm -1 reflected structural modifications in CSNPs compared to native chitosan. Changes in amide bands (Amide I at 1648 cm -1 and Amide II at 1586 cm -1 ) suggested altered hydrogen bonding patterns, influenced by STPP interactions, as shown in Figure 3. Additionally, the presence of P=O stretching vibrations at 2407 cm -1 confirmed the incorporation of phosphate groups from STPP, enhancing nanoparticle stability and cross-linking capabilities. UV-Visible analysis In the UV absorbance graph of chitosan nanoparticles (CNPs), a distinct and sharp peak was observed at 247 nm, indicating the wavelength where these nanoparticles absorbed light most effectively. This peak, characterized by its straight and noise-free shape, as shown in Figure 4, represented specific electronic transitions or absorption bands inherent to the CNPs' molecular structure. The sharpness and clarity of the peak suggested a high degree of uniformity and consistent size distribution of the CNPs. XRD analysis The XRD pattern of chitosan nanoparticles exhibits a prominent broad peak around 2θ = 10°, indicating a significant amorphous character within the material. This broad peak is typical for polymers like chitosan, reflecting the irregular arrangement of the polymer chains. Additionally, the pattern displays several minor peaks between 20° and 80°, which suggest the presence of small crystalline regions or impurities within the predominantly amorphous matrix, as shown in Figure 5. These crystalline regions could be inherent to the semi-crystalline nature of chitosan or may have formed during the nanoparticle preparation process. SEM analysis The scanning electron microscopy (SEM) image of the chitosan nanoparticles revealed a complex morphology characterized by a mixture of amorphous and crystalline structures. At a magnification of 495x and with a scale bar of 10 µm, the nanoparticles showed a rough surface and had an average diameter of 262 nm. The morphology exhibited irregular, amorphous regions alongside smooth crystalline areas, indicating a heterogeneous structure, as shown in Figure 6. The amorphous regions of the nanoparticles display an irregular and non-uniform texture, lacking a defined shape or smoothness. In contrast, the crystalline regions exhibit smooth and well-defined surfaces, suggesting partial crystallization within the chitosan matrix. Assessment of anti-bacterial activity Antibacterial susceptibility test Antibacterial susceptibility tests were conducted to evaluate the effectiveness of different concentrations of maggot crude extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs) against various bacterial strains. The concentrations tested for all the above treatments included 2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), and 10 mg/ml (D). Imipenem 10 mcg (P) was used as a positive control, saline water as a negative control (N), and Ciprofloxacin (5 mcg) (R) was used as a resistance group. These treatments were applied to selected bacterial strains, and their zones of inhibition were measured to determine the antibacterial activity at each concentration. The results obtained after statistical analysis are listed below. Antibacterial activity against E. coli strains Antibacterial activity of the treatments against three strains of E. coli (1874, 1609, and 2021) was measured. From treatment group, MCs at concentration D (10 mg/ml) showed the highest zone of inhibition (21.6 ± 0.6) against E. coli 1874 and (20.3 ± 0.3) E. coli 1609, as shown in Figure 7 and 8, while MEx demonstrated the highest zone of inhibition (20.3 ± 0.3) against E. coli 2021, as shown in Figure 9. The mean values of the zones of inhibition are listed in Table 3. Figure 10 shows the graphical representation of the zones of inhibition of the treatments against the E. coli strains. Antibacterial activity against P. aeruginosa strains Antibacterial activity against two strains of P. aeruginosa (101 and 310) was measured. From treatment groups, MEx at concentration D (10 mg/ml) showed the highest zone of inhibition ( 29.6 ± 0.6; 18.6 ± 0.3) against both P. aeruginosa strains 101 and P310, as shown in Figure 11 and Figure 12. The mean values of the zones of inhibition are listed in Table 4. Figure 13 shows the graphical representation of the zones of inhibition of the treatments against the P. aeruginosa strains. The results indicated that the zones of inhibition increased with the increased in the concentration of the treatments against P. aeruginosa strains . Antibacterial activity against S. aureus strain Antibacterial activity against S. aureus (723) was measured. From treatment group, MEx at concentration D (10 mg/ml) showed the highest zone of inhibition (18.6 ± 0.3), as shown in Figure 14. The mean values of the zones of inhibition are listed in Table 5. Figure 15 shows the graphical representation of the zones of inhibition of the treatments against the S. aureus strain. These results underscore the effectiveness of MEx at higher concentrations in inhibiting the growth of S. aureus . Antibacterial activity against K. pneumoniae strain Antibacterial activity against K. pneumoniae (K310) was measured. From treatment group, MEx at concentration D (10 mg/ml) showed the highest zone of inhibition (2.6 ± 0.3), as shown in Figure 16. The mean values of the zones of inhibition are listed in Table 6. Figure 17 shows the graphical representation of the zones of inhibition of the treatments against the K. pneumoniae strain. These results underscore the effectiveness of MEx at higher concentrations in inhibiting the growth of K. pneumoniae. Statistical analysis of antibacterial activity Statistical analysis of variance (ANOVA) was performed using GraphPad Prism 8.0.2 software, revealing significant variations in zones of inhibition among different treatment groups. The interaction between bacterial strains and treatments was also significant (P < 0.005), suggesting complex relationships influencing antibacterial efficacy across various treatments and bacterial strains. Table 7 illustrates the detailed statistical analysis of antibacterial activity. Morphological and microscopic examination of bacterial strains Identification of Escherichia coli : E. coli is a gram-negative rod shaped and facultative bacteria. It gave pinkish red color on gram staining. It gave greenish fluorescence colonies on EMB (Eosin Methylene Blue) media and pinkish colonies on MacConkey media, as shown in Figure 18. Identification of Klebsiella pneumoniae: Klebsiella pneumoniae is a gram-negative, rod-shaped, and facultative bacterium. It produced a pinkish-red color on Gram staining. On EMB (Eosin Methylene Blue) media, it formed purple colonies usually without metallic green sheen. On MacConkey agar, it produced mucoid, pink colonies due to its ability to ferment lactose, as shown in the Figure 19. Identification of Pseudomonas aeruginosa: Pseudomonas aeruginosa is a gram-negative, rod-shaped, and obligate aerobic bacterium. It produced a pinkish-red color on Gram staining. On EMB (Eosin Methylene Blue) media, it formed colorless colonies because it does not ferment lactose. On MacConkey agar, it also produced colorless colonies, because it is a non-lactose fermenter, as shown in Figure 20. Identification of Staphylococcus aureus : Staphylococcus aureus is a gram-positive, spherical (coccus) bacterium. It gave a purple or blue appearance when observed under a microscope after Gram staining due to its thick peptidoglycan cell wall retaining the crystal violet dye. On blood agar, Staphylococcus aureus produced whitish colonies, as shown in Figure 21. Assessment of antifungal activity Antifungal susceptibility test Antifungal susceptibility tests were conducted to evaluate the effectiveness of different concentrations of maggot crude extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs) against two different molds of Aspergillus. The concentrations tested for all the above treatments included 2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), and 10 mg/ml (D). Ketoconazole (10 µg/mL) was used as a positive control (P), and normal saline water was used as a negative control (N). These treatments were applied to selected fungal molds, and their zones of inhibition were measured to determine the antifungal activity at each concentration. The results obtained after statistical analysis are described below. Antifungal activity against Aspergillus species Antifungal activity against two different molds of Aspergillus ( A. flavus and A. niger ) was measured. From treatment groups, MEx at concentration D (10 mg/ml) showed the highest zone of inhibition ((23.3 ± 0.3; 23 ± 0.5)) against both molds of Aspergillus, as shown in Figure 22 and Figure 23. The mean values of the zones of inhibition are listed in Table 8. Figure 24 shows the graphical representation of the zones of inhibition of the treatments against the Aspergillus species. Statistical analysis of antifungal activity The data was statistically analyzed by using GraphPad Prism 8.0.2 software, ANOVA test was applied to statistically analyze the data. The results of ANOVA analysis of variance examined the zone of inhibition against fungal molds, specifically A. flavus and A. niger , revealed significant effects of different treatments. Significant main effects are observed for treatments (P < 0.005), suggesting effective relationships influencing antifungal efficacy across various treatments and fungal molds. Table 9 describes the detailed results of statistical analysis of antifungal activity. Morphological and microscopic identification of Aspergillus species Identification of Aspergillus flavus : Aspergillus flavus colonies, when grown on SDA (Sabouraud Dextrose Agar) media, exhibited a yellow-green surface. It showed a velvety to cotton-like texture. Under the microscope, Aspergillus flavus showed septate, hyaline hyphae and long, unbranched conidiophores ending in spherical vesicles covered with phialides. The rough-walled conidia were spherical, as shown in Figure 25. Identification of Aspergillus niger : Aspergillus niger colonies exhibited a white base with black colonies on the surface of SDA (Sabouraud Dextrose Agar) media. It showed a granular to powdery texture. Microscopic examination with lactophenol cotton blue staining revealed septate, clear hyphae and long, smooth conidiophores ending in large, round vesicles. The conidia were in large, radiating chains, forming a dense, black powdery mass, as shown in Figure 26. DISCUSSION The biomedical field has seen a growing interest in the use of natural and biocompatible materials for therapeutic applications. Among these, maggot extract, chitosan, and its derivative products have emerged as effective options. Their unique properties, including biocompatibility, biodegradability, and potent bioactivity, make them attractive for various therapeutic purposes. This study evaluated the antimicrobial activity using the disc diffusion method. All treatments demonstrated positive antimicrobial effects. However, there were certain differences in efficacy among the treatments for each group. In this study, the substantial reduction during the production of chitosan was observed which is in accordance with the previous research [ 52 – 54 ]. The degree of deacetylation of chitosan was determined to be 90.57%, is comparable to values reported by [ 50 ], where deacetylation levels typically range from 85–95% for effective chitosan preparation. The stability of the chitosan nanoparticles pellet prepared by the ion-gelatin method at a 0.4% chitosan concentration and the sedimentation problem at higher concentrations align with findings from [ 55 ], who reported similar challenges in nanoparticle preparation related to chitosan concentration. In the current findings, the GC-MS analysis of the maggot crude extract (MEx) identified twenty distinct compounds with diverse bioactive properties. The identified compounds include alcohols, esters, cyclic lactones, fatty acid derivatives, phthalate and n-octane, which have shown promising biological activities in previous studies. For instance, the antioxidant properties of compounds like 3-methyl-1-butanol (isoamyl alcohol) and isovaleric acid isopentyl ester are consistent with the previous findings [ 56 ], who reported similar compounds exhibiting significant antioxidant effects. Additionally, the presence of cyclic lactones and fatty acid derivatives, including oleic acid, highlighted potential neuroprotective properties as demonstrated by previous literature [ 57 ], who reported the neuroprotective effects of oleic acid. Diethyl phthalate stands out for its anti-inflammatory properties. Previous research [ 58 ] also demonstrated the anti-inflammatory effects of similar compounds, including phthalates. Furthermore, the antibacterial activity of n-octane aligns with the research of [ 59 ]. In this research work, the characterization of the CNPs revealed key structural details. The FTIR spectra revealed intensified O-H and NH 2 stretching vibrations and the presence of P = O stretching, indicating successful cross-linking with sodium tripolyphosphate (STPP), which supports the findings of [ 47 ], who reported that STPP enhances nanoparticle stability and cross-linking, as well as [ 48 ], who observed similar effects of STPP on nanoparticle properties. The UV-Visible analysis of the prepared chitosan nanoparticles was identified a sharp peak at 247 nm that indicated the presence of CO group as analyzed by [ 60 ]. XRD analysis of the current study revealed a broad peak around 2θ = 10°, indicative of the amorphous nature of chitosan nanoparticles, with minor peaks suggesting some degree of crystalinity. This finding is consistent with previous studies by [ 61 – 62 ], who examined an amorphous structure with minor crystalline regions in chitosan nanoparticles. SEM analysis further confirmed the heterogeneous morphology of the nanoparticles, revealing a mixture of amorphous and crystalline structures with an average particle size of 262 nm. These observations are in accordance with the findings of [ 63 ], who reported similar structural characteristics in chitosan nanoparticles. In the present study, the results of the antibacterial susceptibility tests demonstrated notable efficacy of maggot crude extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs) against various bacterial strains. MEx exhibited the highest antibacterial activity against E. coli (2021), P. aeruginosa (101 and 310), S. aureus (723), and K. pneumoniae (310) strains, with the greatest zones of inhibition observed at the highest concentration (10 mg/ml). This finding aligns with previous studies by [ 64 – 65 ], who demonstrated the strong antimicrobial properties of crude maggot extract. Recent literature [ 66 ] examined the compounds within MEx and found them to be particularly effective against the tested bacterial strains. Similarly, MCs showed promising results against all tested bacterial strains, with particularly strong antibacterial effects at higher concentrations against E. coli (1874 and 1609). This is consistent with the research findings of [ 67 ], who indicated that chitosan-based materials have potent antibacterial effects due to their ability to disrupt microbial cell membranes. CNPs also exhibited antibacterial activity against all the mentioned strains, but their effectiveness was comparatively less than that of MEx and MCs. However, according to [ 68 ], chitosan nanoparticles showed the highest antibacterial activity. In the present findings, E. coli was identified by its characters like greenish fluorescence on Eosin Methylene Blue (EMB) media and pinkish colonies on MacConkey media. These observations align with the work of [ 69 ], who described similar characteristics for E. coli . [ 70 ] demonstrated the ability of E. coli to ferment lactose, producing acid that results in color changes on these media. Klebsiella pneumoniae was also confirmed by the formation of mucoid, pink colonies on MacConkey agar and purple colonies on EMB media without a metallic green sheen. These results were in accordance with previous studies by [ 71 ], who reported similar colony morphologies and fermentation profiles for K. pneumoniae . Pseudomonas aeruginosa , a non-lactose fermenter, formed colorless colonies on both EMB and MacConkey media, and appeared pinkish-red on Gram staining. These observations are in consistent with the findings of [ 72 ]. Staphylococcus aureus displayed its characteristic gram-positive, spherical morphology, with a purple or blue appearance on Gram staining due to the retention of crystal violet dye in its thick peptidoglycan cell wall. The production of whitish colonies on blood agar is indicative of its ability to grow on enriched media and form distinctive colonies. These findings are in line with the study by [ 73 ], which described similar staining characteristics and colony morphology for S. aureus . In the current analysis, the antifungal susceptibility tests showed significant differences in the effectiveness of various treatments against Aspergillus flavus and Aspergillus niger . Maggot crude extract (MEx) at the highest concentration (10 mg/ml) had the most notable antifungal activity, with inhibition zones of 23.3 ± 0.3 mm for A. flavus and 23 ± 0.5 mm for A. niger . These results highlight the strong potential of MEx as an antifungal agent. This observation is in line with previous research by [ 74 ], which supports these findings. Maggot chitosan (MCs) showed impressive antifungal activity, with the highest concentration (10 mg/ml) yielding inhibition zones of 20 ± 0.5 mm for both A. flavus and A. niger . Similarly, chitosan nanoparticles (CNPs) were effective, demonstrating inhibition zones of 22.6 ± 0.3 mm for A. flavus and 18.6 ± 0.3 mm for A. niger at the same concentration. These findings support the previous study by [ 75 ], who also found that the increased surface area and reactivity of chitosan nanoparticles enhance their effectiveness against fungal strains. In this research, Aspergillus flavus formed yellow-green colonies on Sabouraud Dextrose Agar (SDA) with a velvety to cotton-like texture. The observed microscopic structures were septate, hyaline hyphae, long unbranched conidiophores ending in spherical vesicles, and rough-walled spherical conidia. These findings are in line with those by [ 76 ], who also noted the distinctive yellow-green pigmentation and conidial structures of A. flavus . On the other hand, Aspergillus niger produced colonies with a white base and black surface on SDA, displaying a granular to powdery texture. Microscopic examination showed septate, clear hyphae, long smooth conidiophores with large round vesicles, and dense, black conidial heads. This matches the descriptions by [ 77 ], who highlighted the characteristic black pigmentation and granular colony texture of A. niger , along with its conidial arrangement. CONCLUSION The findings underscore the potential of MEx, MCs, and CNPs as natural and biopolymer-based treatments as effective alternatives to traditional treatments, demonstrating significant benefit in antimicrobial effects suggest promising applications in therapeutic settings, offering new avenues for enhancing patient care and treatment outcomes. These treatments not only exhibit strong efficacy in inhibiting bacterial and fungal infections. Future research should focus on elucidating the specific mechanisms underlying their therapeutic effects, optimising their formulations for clinical use, and exploring their potential for treating a broader range of medical conditions. Declarations I, MAIRA MUNIR, hereby declare that this manuscript titled “ Comparative analysis of antimicrobial properties of maggot ( Musca domestica ) crude extract, maggot chitosan and chitosan nanoparticles ” is an original work, and it has not been submitted or published elsewhere. All data and findings presented in this manuscript are accurate and have been obtained through ethical research practices. I confirm that there are no conflicts of interest related to this manuscript. I have adhered to the ethical guidelines and standards for research. The study has received the necessary ethical approval from Lahore College for Women University, Lahore, Pakistan, ss per guidelines of OCCUPATIONAL AND HEALTH ACT, 2007, and follows the protocols set forth by the relevant ethical review boards. Research Ethical Review Committee Prof. Dr. Farzana Rashid Prof. Dr. Saima Sharif Dr. Shagufta Naz (Associate Professor) Dr. Ghazala Jabeen (Associate Professor) Dr. Sumera Sajjad (Associate Professor) Consent to Publish Additionally, I confirm that all authors listed in this paper have contributed to the research, and their consent has been obtained for publication. The authors have no conflict of interest to declare. Funding This research did not receive any grant from any funding agencies. Data Availability All data supporting the findings of this study are available within the paper. I declare that this manuscript is my original work. 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Isolation and identification of Aspergillus niger from the onions in Bharuch, Gujarat, India. Adv Microbiol Res. 2024;5(1):178–80.Top of Form Tables Table 1: Summarized results of GC-MS analysis of maggot crude extract (MEx). Sr.No. RT Compound Name Area Area% Molecular Weight Molecular Formula 1 2.409 3-methyl-1-butanol (isoamyl alcohol) 1285204 30.72 88 C 5 H 12 O 2 2.975 n-Octane 71570 1.71 114 n-C 8 H 18 3 6.431 1-(5-Bicyclo [2.2.1] heptyl) ethylamine 51030 1.22 139 C 9 H 17 N 4 7.541 n-Pentanal or Valeraldehyde 31265 0.75 86 C 5 H 10 O 5 7.646 Isovaleric acid, Isopentyl ester. 300577 7.18 172 C 10 H 20 O 2 6 12.019 Furfuryl alcohol, tetrahydro- 88655 2.12 102 C 5 H 10 O 2 7 14.765 Diethyl phthalate 494809 11.83 222 C 12 H 14 O 4 8 18.542 Z,Z-8,10-hexadecadien-1-ol 49884 1.19 238 C 16 H 30 O 9 18.948 9-Octadecenoic acid (Z)-, methyl ester 40653 0.97 296 C 19 H 36 O 2 10 20.286 Oleic acid (9Z)-, 9-octadecenoic acid (Z)- 294803 7.05 282 C 18 H 34 O 2 11 20.346 Dimethoxybicyclo [3.3.1]nona-2,4-dione 460266 11 212 C 11 H 16 O 4 12 22.249 9,12-Octadecadienoic acid (Z,Z)- 45124 1.08 280 C 18 H 32 O 2 13 22.297 Z,Z-3,13-Octadecadien-1-ol (4Z,13Z)- 29522 0.71 266 C 18 H 34 O 2 14 23.429 Z,Z-2,13-Octadecadien-1-ol (2Z,13Z)- 25609 0.61 266 C 18 H 34 O 2 15 23.85 Z,Z-3,13-Octadecadien-1-ol (4Z,13Z)- 84265 2.01 266 C 18 H 34 O 2 16 23.94 Z,Z-2,13-Octadecadien-1-ol (2Z,13Z)- 89464 0.7 266 C 18 H 34 O 2 17 24.002 Z,Z-2,13-Octadecadien-1-ol (2Z,13Z)- 493430 11.79 266 C 18 H 34 O 2 18 24.265 6-Octadecenoic acid, methyl ester (Z)- 193460 4.62 296 C 19 H 36 O 2 19 24.527 6-Octadecenoic acid, methyl ester (Z)- 70025 1.67 296 C 19 H 36 O 2 20 25.755 Oleic acid, 9-octadecenoic acid (Z)- 44280 1.06 282 C 18 H 34 O 2 Table 2: Summarized results of chitosan prepared from Musca domestica. Process Material obtained Dry weight (grams) Defatting Defatted crude chitin 88.6 Deproteinization Deproteinized crude chitin 9.36 Demineralization Chitin 5.9 Deacetylation Chitosan 3.10 Decolorization Decolorized Chitosan 2.55 Table 3: Antibacterial activity of treatments against E. coli strains. Groups Zones of inhibition in mm (mean ± SE) against E. coli strains E. coli (1874) E. coli (1609) E. coli (2021) Control Positive Control 23.6 ± 0.3 42.6 ± 0.6 41 ± 0.5 Negative Control 0 0 0 Resistance 0 0 0 Treatment Groups Concentrations of MEx A 11 ± 0.5 12.6 ± 0.3 15.6 ± 0.8 B 13.6 ± 0.6 14.3 ± 0.8 19.3 ± 0.8 C 16.6 ± 0.6 15.6 ± 0.3 18.3 ± 0.3 D 14.6 ± 0.8 17.6 ± 0.3 20.3 ± 0.3 Concentrations of MCs A 11.6 ±0.3 14 ± 0.5 13 ± 0.5 B 12.6 ± 0.3 18 ± 0.5 12 ± 0.5 C 14.6 ± 0.6 16 ± 0.5 14 ± 0.5 D 21.6 ± 0.6 20.3 ± 0.3 18 ± 0.5 Concentrations of CNPs A 8.3 ± 0.3 11.6 ± 0.3 7.3±0.3 B 9 ± 0.5 8.3 ± 0.8 9.3 ± 0.3 C 9.3 ± 0.3 9.3 ± 0.3 12.6 ± 0.8 D 9.5 ± 0.2 12 ± 0.5 13.6 ± 0.3 Table 4: Antibacterial activity of treatments against P. aeruginosa strains. Groups Zones of inhibition in mm (mean ± SE) against P. aeruginosa strains Control Groups P. aeruginosa (101) P. aeruginosa (310) Positive Control 44.3 ± 0.6 37.3 ± 0.8 Negative Control 0 0 Resistance 0 0 Treatment Groups Concentrations of MEx A 12.6 ± 0.3 9.3 ± 0.3 B 18.6 ± 0.3 10.6 ± 0.3 C 23.3 ± 0.6 12.6 ± 0.3 D 29.6 ± 0.6 18.6 ± 0.3 Concentrations of MCs A 15.3 ± 0.3 10.6 ± 0.3 B 13 ± 1 11 ± 0.5 C 15.6 ± 0.8 13 ± 0.5 D 17.6 ± 0.8 17 ± 0.5 Concentrations of CNPs A 11 ± 0.5 10 ± 0.5 B 16 ± 0.5 11.14 ± 0.8 C 11.67 ± 0.6 11.3 ± 0.8 D 12.6 ± 0.3 12.6 ± 0.6 Table 5: Antibacterial activity of treatments against S. aureus . Groups Zones of inhibition in mm (mean ± SE) against S. aureus Control Groups S. aureus (723) Positive Control 42.3 ± 0.6 Negative Control 0 Resistance 0 Treatment Groups Concentrations of MEx A 11 ± 0.5 B 14 ± 0.5 C 18.3 ± 0.3 D 18.6 ± 0.3 Concentrations of MCs A 10 ± 0.5 B 8.6 ± 0.3 C 13 ± 0.5 D 17.3 ± 0.6 Concentrations of CNPs A 8.3 ± 0.3 B 11.3 ± 0.3 C 11 ± 0.5 D 9.3 ± 0.6 Table 6: Antibacterial activity of treatments against K. pneumoniae . Groups Zones of inhibition in mm (mean ± SE) against K. pneumoniae Control Groups K. pneumoniae (310) Positive Control 41.6 ± 0.8 Negative Control 0 Resistance 0 Treatment Groups Concentrations of MEx A 11.6 ± 0.6 B 16.3 ± 0.3 C 18.3 ± 0.3 D 22.6 ± 0.3 Concentrations of MCs A 12.3 ± 0.3 B 15.3 ± 0.8 C 13 ± 0.5 D 17.3 ± 0.8 Concentrations of CNPs A 9.3 ± 0.8 B 8.6 ± 0.3 C 11.3 ± 0.8 D 12.6 ± 0.6 Table 7: Statistical analysis of antibacterial activity. ANOVA test Sum of square (SS) Degree of freedom (DF) Mean square (MS) F P value Row Factor 7994 14 571 82.67 P < 0.005 Column Factor 190.7 6 31.78 4.601 P < 0.005 Residual 580.2 84 6.907 Table 8: Antifungal activity of treatments against Aspergillus species. Groups Zones of inhibition in mm (mean ± SE) against Aspergillus molds Control Groups A. flavus A. niger Positive Control 29.6 ± 0.6 35.3 ± 0.3 Negative Control 0 0 Treatment Groups Concentrations of MEx A 15 ± 0.5 14 ± 0.5 B 17.3 ± 0.8 15 ± 0.5 C 18 ± 0.5 17.3 ± 0.8 D 23.3 ± 0.3 23 ± 0.5 Concentrations of MCs A 15 ± 0.5 15 ± 0.5 B 16 ± 0.5 15.6 ± 0.6 C 18 ± 0.5 17 ± 0.5 D 20 ± 0.5 20 ± 0.5 Concentrations of CNPs A 13 ± 0.5 11.3 ± 0.3 B 15.3 ± 0.3 15 ± 0.5 C 16 ± 0.5 16.6 ± 0.3 D 22.6 ± 0.3 18.6 ± 0.3 Table 9: Statistical analysis of antifungal activity. ANOVA test Sum of square (SS) Degree of freedom (DF) Mean square (MS) F P value Row Factor 1788 14 127.7 61.74 P<0.005 Column Factor 0.972 1 0.972 0.4699 P<0.005 Residual 28.96 14 2.068 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-6084200","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":424262172,"identity":"42489c7c-55c5-45a8-9ddb-eb4d3a6f3f7c","order_by":0,"name":"Maira Munir","email":"","orcid":"","institution":"Anhui University","correspondingAuthor":false,"prefix":"","firstName":"Maira","middleName":"","lastName":"Munir","suffix":""},{"id":424262173,"identity":"9576c494-2ac6-4094-905c-27af5e6e2644","order_by":1,"name":"Saffora 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chitosan nanoparticles.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/74cfb840d6f0382aed528924.png"},{"id":77859774,"identity":"ad39f9b8-d784-4067-a388-5c6f7dd0d4f0","added_by":"auto","created_at":"2025-03-06 08:33:55","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":10929,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eUV-visible graph of chitosan nanoparticles.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/7d8728d6b36d090520b895db.png"},{"id":77859773,"identity":"f7bd93ea-139c-4719-a900-a60679981698","added_by":"auto","created_at":"2025-03-06 08:33:51","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":11738,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eXRD graph of chitosan nanoparticles.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/1d7f22a7de0b44a2b32e01d3.png"},{"id":77861040,"identity":"e0b54250-3c07-4cd3-a667-c00c5fd9abb3","added_by":"auto","created_at":"2025-03-06 08:41:46","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":392700,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMorphology of chitosan nanoparticles by SEM analysis.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig6.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/640b801e926791b1c058b6df.png"},{"id":77861226,"identity":"2c129e89-3eff-41a1-8f10-74d9a3b8062c","added_by":"auto","created_at":"2025-03-06 08:49:46","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1804643,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntibacterial activity against strains of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eE. coli \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e(1874). (a) Positive control (P), negative control (N), and resistant group (R); (b) Concentrations of MEx; (c) Concentrations of MCs; (d) Concentrations of CNPs.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig7.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/a149f4cd4b8d44bbee1a237d.png"},{"id":77861022,"identity":"760de3b9-ade5-4976-bab7-6a16fe765ed2","added_by":"auto","created_at":"2025-03-06 08:41:45","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":1872738,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntibacterial activity against strains of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eE. coli \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e(1609). (a) Positive control (P), negative control (N), and resistant group (R)); (b) Concentrations of MEx; (c) Concentrations of MCs; (d) Concentrations of CNPs.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig8.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/24b28666d72c856063f0fa7e.png"},{"id":77861043,"identity":"cdd63bf5-43a6-4c60-a892-fab9b9091d35","added_by":"auto","created_at":"2025-03-06 08:41:46","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":1766489,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntibacterial activity against strains of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eE. coli \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e(2021). (a) Positive control (P), negative control (N), and resistant group (R)); (b) Concentrations of MEx; (c) Concentrations of MCs; (d) Concentrations of CNPs.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig9.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/b9f7dd3f831eedbdfbd3fb2f.png"},{"id":77861052,"identity":"e5210580-a2ef-4f59-8574-2c0c9a551b1f","added_by":"auto","created_at":"2025-03-06 08:41:47","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":24223,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical representation of antibacterial activity of treatments against \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eE.coli \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003estrains.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig10.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/8249f5d961fe8a4b7db0d089.png"},{"id":77861039,"identity":"5fbda9f7-776b-4292-aa39-64682d279938","added_by":"auto","created_at":"2025-03-06 08:41:45","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":1478316,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntibacterial activity against strains of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePseudomonas aeruginosa\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e (101). (a) Antibacterial effects of positive control (P), negative control (N), and resistant group (R); (b) Concentrations of MEx; (c) Concentrations of MCs; (d) Concentrations of CNPs.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig11.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/59872dd314a491c9a56840c1.png"},{"id":77861049,"identity":"3c328d39-c8fa-4cba-8b00-7ea851af9bc2","added_by":"auto","created_at":"2025-03-06 08:41:47","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":1469822,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntibacterial activity against strains of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePseudomonas aeruginosa\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e (310). (a) Antibacterial effects of positive control (P), negative control (N), and resistant group (R); (b) Concentrations of MEx; (c) Concentrations of MCs; (d) Concentrations of CNPs.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig12.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/7ceaf77f5116176af72dc933.png"},{"id":77859760,"identity":"0152c86c-2c28-407d-8a74-8815e76c5f2e","added_by":"auto","created_at":"2025-03-06 08:33:49","extension":"png","order_by":13,"title":"Figure 13","display":"","copyAsset":false,"role":"figure","size":22668,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical representation of antibacterial activity of treatments against \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eP. aeruginosa\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e strains.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig13.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/8ce1aa204bcac401b3632d79.png"},{"id":77861046,"identity":"80b3f683-ccdf-4aec-825c-2b7709dae983","added_by":"auto","created_at":"2025-03-06 08:41:46","extension":"png","order_by":14,"title":"Figure 14","display":"","copyAsset":false,"role":"figure","size":1546902,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntibacterial activity against strains of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eStaphylococcus aureus.\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e (a) Antibacterial effects of positive control (P), negative control (N), and resistant group (R); (b) Concentrations of MEx; (c) Concentrations of MCs; (d) Concentrations of CNPs.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig14.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/1811fad4c3efec93c526f220.png"},{"id":77859748,"identity":"cc96feff-0988-4f35-a796-50fbfd319afa","added_by":"auto","created_at":"2025-03-06 08:33:48","extension":"png","order_by":15,"title":"Figure 15","display":"","copyAsset":false,"role":"figure","size":20986,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical representation of antibacterial activity of treatments against \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eS. aureus\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e strain.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig15.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/49e685b9564371b780094a0d.png"},{"id":77859708,"identity":"42b82bdc-499b-45ee-81e1-f84203e535f8","added_by":"auto","created_at":"2025-03-06 08:33:46","extension":"png","order_by":16,"title":"Figure 16","display":"","copyAsset":false,"role":"figure","size":1564892,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntibacterial activity against strains of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eKlebsiella pneumoniae\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. (a) Antibacterial effects of positive control (P), negative control (N), and resistant group (R); (b) Concentrations of MEx; (c) Concentrations of MCs; (d) Concentrations of CNPs.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig16.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/cb003ed5f22bcde872e45220.png"},{"id":77859693,"identity":"6976d628-fa36-47ca-b097-7ff82cd768fe","added_by":"auto","created_at":"2025-03-06 08:33:45","extension":"png","order_by":17,"title":"Figure 17","display":"","copyAsset":false,"role":"figure","size":17886,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical representation antibacterial activity of treatments against \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eK. pneumoniae \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003estrain.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig17.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/95b9ab1201aa0869009c2257.png"},{"id":77859703,"identity":"f39f1fd1-fd7b-42e1-94b1-a83aefea5444","added_by":"auto","created_at":"2025-03-06 08:33:46","extension":"png","order_by":18,"title":"Figure 18","display":"","copyAsset":false,"role":"figure","size":1287871,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMorphological and microscopic identification of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eE. coli\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. A: Greenish colonies of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eE. coli\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e on Eosin Methylene Blue (EMB) media; B: Pink colonies of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eE. coli\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e on MacConkey media; C: Microscopic examination of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eE. coli\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e after Gram staining.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig18.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/a4eecb0e5b8dbcdffcfe6c0e.png"},{"id":77859759,"identity":"5a6d2b37-83f5-4b8f-a4c3-91b3a4ebc9c9","added_by":"auto","created_at":"2025-03-06 08:33:48","extension":"png","order_by":19,"title":"Figure 19","display":"","copyAsset":false,"role":"figure","size":1263689,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMorphological and microscopic identification of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eKlebsiella pneumoniae\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. A: mucoid pinkish colonies of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eK. pneumoniae\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e on EMB media; B: purplish colonies of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eK. pneumoniae\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e on MacConkey media; C: microscopic examination of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eK. pneumoniae\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon gram staining.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig19.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/ce79af65aaeda09106b11617.png"},{"id":77861063,"identity":"19a78258-7114-40d1-a63f-b69d27374a43","added_by":"auto","created_at":"2025-03-06 08:41:49","extension":"png","order_by":20,"title":"Figure 20","display":"","copyAsset":false,"role":"figure","size":1100562,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMorphological and microscopic identification of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ePseudomonas aeruginosa\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. A: colorless colonies of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eP. aeruginosa\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e on EMB media; B: colorless colonies of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eP. aeruginosa\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e on MacConkey media; C: microscopic examination of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eP. aeruginosa \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon gram staining.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig20.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/7809d04d4b4867bd587f45d0.png"},{"id":77859722,"identity":"8dfae268-9758-4a09-8048-bcca631717de","added_by":"auto","created_at":"2025-03-06 08:33:46","extension":"png","order_by":21,"title":"Figure 21","display":"","copyAsset":false,"role":"figure","size":745257,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMorphological and microscopic identification of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eStaphylococcus aureus\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. A: whitish colonies of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eS. aureus\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e on Blood agar; B: microscopic examination of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eS. aureus\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e after gram staining.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig21.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/6c085fc85e4f0a7aeefb5cf0.png"},{"id":77861047,"identity":"497c382f-77b3-4005-aff2-0029fadb9e5e","added_by":"auto","created_at":"2025-03-06 08:41:46","extension":"png","order_by":22,"title":"Figure 22","display":"","copyAsset":false,"role":"figure","size":1784122,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntifungal activity against \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAspergillus flavus\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. (a) Antifungal effects of positive control (P) and negative control (N); (b) Concentrations of MEx; (c) Concentrations of MCs; (d) Concentrations of CNPs.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig22.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/decc282492947986324a02d6.png"},{"id":77859745,"identity":"7145fd00-02e4-40dd-a941-2ee84f5eb0db","added_by":"auto","created_at":"2025-03-06 08:33:48","extension":"png","order_by":23,"title":"Figure 23","display":"","copyAsset":false,"role":"figure","size":1324242,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAntifungal activity against \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAspergillus niger\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. (a) Antifungal effects of positive control (P) and negative control (N); (b) Concentrations of MEx; (c) Concentrations of MCs; (d) Concentrations of CNPs.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig23.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/e3d3696433f37dbd3654e5c2.png"},{"id":77859730,"identity":"95fe41a8-4717-4616-8d43-a698b6fc07b3","added_by":"auto","created_at":"2025-03-06 08:33:47","extension":"png","order_by":24,"title":"Figure 24","display":"","copyAsset":false,"role":"figure","size":22791,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical representation of antifungal activity of treatments against Aspergillus species.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig24.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/970f7aec82e447b75fa8e394.png"},{"id":77859736,"identity":"31af604c-5c07-4c1c-9326-c4e57e6fba8f","added_by":"auto","created_at":"2025-03-06 08:33:47","extension":"png","order_by":25,"title":"Figure 25","display":"","copyAsset":false,"role":"figure","size":835411,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eIdentification of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAspergillus flavus\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. A: On SDA media; B: Microscopic examination.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig25.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/fa657117e4d2577e0e1e9992.png"},{"id":77859738,"identity":"d3d339cb-ed5e-4e61-a358-cd60534aa335","added_by":"auto","created_at":"2025-03-06 08:33:47","extension":"png","order_by":26,"title":"Figure 26","display":"","copyAsset":false,"role":"figure","size":944433,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eIdentification of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAspergillus niger\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e. A: On SDA media; B: Microscopic examination.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig26.png","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/48762eb67ef1b50f6d1860cd.png"},{"id":79548389,"identity":"bbab4548-79e3-4054-8998-f1f6c9d0063c","added_by":"auto","created_at":"2025-03-31 06:17:41","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":33740777,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6084200/v1/b56d2b48-3e27-4f1e-abca-8657f66be9a4.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Comparative analysis of antimicrobial properties of maggot (Musca domestica) crude extract, maggot chitosan and chitosan nanoparticles","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eMaggots are the larval stage of the \u003cem\u003eMusca domestica\u003c/em\u003e species that belong to the Diptera: Muscidea family. The common housefly is acknowledged for its role as a sanitation pest. However, because of the short life span, high potential for reproduction, and efficient breakdown of organic waste, it has also been emerged as a valuable source of protein, chitin, and chitosan [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Housefly larvae have been a source for the extraction of valuable materials like peptides, chitosan, phospholipids, and antibiotics. Based on initial observations, it was determined that roughly 55% of dried housefly larvae's composition is protein, 9.1% is crude chitin, and 8.8% is fat [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHousefly larvae are well-known for their non-toxic properties. They have been clinically used to treat various conditions, such as ecthyma, wounds, and bacterial infections [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Due to the ease of access and low cost associated with breeding \u003cem\u003eMusca domestica\u003c/em\u003e, it has been proposed as a potential alternative to \u003cem\u003eLucilia sericata\u003c/em\u003e for this therapy [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Maggot therapy, widely known for its powerful ability to fight off different infections and emerges as the fastest and most effective method for clearing necrotic tissue and treating infections [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Maggots have been observed to reduce the amount of bacteria in wounds due to their multiple antibacterial actions [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Maggot excretions can terminate several pro-inflammatory reactions by neutrophils, including degranulation, chemotaxis, respiratory burst, and integrin expression [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eChitinous materials such as chitin and chitosan are among the planet's second-most abundant biopolymers, following cellulose. Their versatility spans over two hundred applications, ranging from agriculture to biomedicine, cosmetics, food, and textiles, show potential as industrial effluent refiners and chelating agents [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Chitin mainly comes from the shells of crustaceans such as shrimp and crabs [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. However, these shells have a lot of calcium, wax, and pigment, which makes it expensive to extract chitosan from them. Comparatively, housefly larvae make chitin extraction easier when contrasted with shrimp or crabs [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], as they have reduced amounts of ash, crude fat, and crude protein [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eChitosan is obtained from the process of deacetylation of chitin, a process that removes acetate groups. The shells of housefly larvae are rich in chitin, making them a significant source of chitosan [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. The depolymerization of chitosan has significant interest as it enhances the water solubility of the resulting products. Furthermore, numerous chemical alterations have been investigated to enhance the solubility of chitosan and broaden its potential applications [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Chitosan and its oligosaccharides offer a lot of benefits, such as fighting tumors, protecting the nervous system, combating fungal infections, and reducing inflammation [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. In recent years, chitin, chitosan, and their derivatives have gained significant attention for their antimicrobial properties against diverse microorganisms, including bacteria, yeast, and fungi [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Chitosan shows its ability to fight bacteria best in acidic conditions because it doesn't dissolve well above pH 6.5 [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. It is more effective and has increased efficacy in combating gram-positive bacteria in comparison to gram-negative ones [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe antibacterial and antifungal profile of chitosan and its derivatives have made them significant components in commercial disinfectants [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Chitosan has abundant advantages over other disinfectants because it is highly effective against bacteria and less harmful to mammalian cells [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. The antimicrobial activity of chitosan is the most significant bioactivity and it has been used in the development of biomedical materials as well as in improving the functionality of other polymeric materials such as fibers and food preservation [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Chitin and chitosan, both, have the ability to stimulate the host's defense system and protect the host against pathogen invasion [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Two main mechanisms have been proposed for how chitosan inhibits microbial cells. One is a positive charge present on chitosan that disrupts bacterial metabolism by sticking to the surface of bacterial cells [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Another reason is that chitosan adsorbs, preventing the transcription of RNA from DNA by binding to DNA molecules [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eChitosan is an ideal material for enzyme immobilization because of its enhanced resistance to chemical degradation and the ability to prevent metal ions from interfering with enzymes [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. The amino functional group found in chitosan is suitable for immobilizing enzymes [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Research on the pro- and anti-inflammatory properties of chitosan and its derivatives revealed that they suppress the development of colitis and increase the induction of the anti-inflammatory IL-10 cytokine in animal blood [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eNanotechnology has gained prominence in several fields because of its unique qualities and the growing problem of microbial drug resistance. Nanoparticles (NPs) can interact with biological molecules and, most importantly, microorganisms due to their unique physical and chemical properties that allow them to pass through barriers [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Chitosan nanoparticles (CNPs) combine the qualities of nanoparticles with those of chitosan, including surface and interface effects, small size, and quantum size effects [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. The surface-to-volume ratio of chitosan is higher at the nanoscale, which increases its affinity for bacteria and fungi. Chitosan nanoparticles directly interfere with microbial cell membranes, cell walls, key proteins, and enzymes, capable of inhibiting pathogen growth and inducing cell death through mechanisms distinct from conventional antibiotics, which may have developed resistance. The size, shape, and chemical properties of nanoparticles can be manipulated to enhance these molecular interactions, optimizing their overall effectiveness [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Consequently, a variety of medications can be produced using nanoparticle manufacturing techniques [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eChitosan-based nanoparticles show potential in medicine and the food industry because materials that are nanoscale in size acquire unique characteristics that set them apart from bulk materials and isolated atoms [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e], and are employed in the control of ocular infections, gastrointestinal disorders, lung disorders, cancer, and drug delivery to the brain [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. When chitosan comes into contact with anions, it has the ability to form gel and beads. Due to this attribute, it can be used to deliver drugs [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. The development of a wide range of colloidal delivery vehicles has been made possible by the potential use of CNPs as carriers [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. In biological media, chitosan nanoparticles (CNPs) can withstand biological barriers and prevent macromolecules from degradation. It can also deliver medicinal products or macromolecules to a target location by controlled release [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eSmall-sized CNPs also contribute to its efficiency in interfacial interaction with cell membranes because the tiny particles are taken up by the cell through endocytosis [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. CNPs can increase the bioavailability of pharmaceutical products by altering the pharmacokinetics and insulating the encapsulated drugs [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. In a dose-dependent manner, chitosan nanoparticles can effectively cause human keratinocytes and monocytes to produce cytokines [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. CNPs are an excellent environmentally friendly material with non-toxic bioactive and natural materials with exceptional physicochemical, antimicrobial, and biological qualities [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eCNPs have been studied extensively in the fields of biomedical materials and tissue engineering because of their good water solubility, biocompatibility, biodegradability, antimicrobial, film forming properties, hemostasis, wound healing, and anti-inflammatory effects. They were successfully used as a drug delivery system. Target therapy may benefit from the use of these NPs [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eRecent studies underscore the potential of maggot therapy and chitosan-based treatments for different infection control. The efficacy of maggot extract (MEx) and chitosan in enhancing wound healing and exhibiting antimicrobial properties [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. Maggot chitosan (MCs) and chitosan nanoparticles (CNPs) are effective against various bacterial strains [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. Additionally, The potential of chitosan nanoparticles in biomedical applications, which is supported by your UV analysis showing specific absorbance characteristics for CNPs [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe study aimed to compare the antimicrobial properties of maggot extract, maggot chitosan, and chitosan-based nanoparticles. The objectives were: to extract chitin from \u003cem\u003eMusca domestica\u003c/em\u003e for chitosan preparation, to synthesize and characterize chitosan nanoparticles using UV-visible spectroscopy, XRD, FTIR, and SEM, and to evaluate and compare the antimicrobial effectiveness of maggot extract and chitosan nanoparticles against \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e, \u003cem\u003eStaphylococcus aureus\u003c/em\u003e, and \u003cem\u003eKlebsiella pneumoniae\u003c/em\u003e.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003eThe experiment was conducted in the Entomology Lab of the Department of Zoology at Lahore College for Women University, Lahore.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMaggot collection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA housefly breeding site was established in a plastic jar. Flies were allowed to feed on chicken offal. \u0026nbsp;The breeding sites were left undisturbed for a week, allowing the flies to lay eggs, hatch, and develop into larvae or maggots. During this period, the breeding site was regularly monitored to check the optimal environmental factors, such as temperature and moisture levels, to ensure ideal conditions for maggot growth. After 7 to 8 days, the breeding sites were carefully examined, and the mature maggots, second and third larval instars were carefully collected. After the collection, maggots were killed by placing them in hot water at 80\u0026deg;C for 3 to 5 minutes and then washed with saline water to remove any debris. The maggots were dried in an air-dried oven at 55\u0026deg;C. Once dried, the maggots were stored in airtight containers or sealed bags at room temperature for future use.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of crude extract of maggots\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe crude extract of maggots was prepared following the methodology of\u0026nbsp;[6], with some modifications. About two hundred grams of maggots were selected and crushed in a mortar and pestle. The crushed maggots were added to a conical flask and two volumes of 10X Tris NaCl EDTA (TNE) buffer (pH 7.5-8) precooled at 4\u003csup\u003eo\u003c/sup\u003eC were added. The mixture was homogenized for 15 minutes at room temperature. After sufficient homogenization, the conical flask was covered with aluminum foil and it was left at room temperature for 24 hours.\u003c/p\u003e\n\u003cp\u003eThe next day, the homogenate was transferred to a sterile falcon tube and centrifuged in a refrigerated centrifugal machine (MSE Harrier 18/80) at 6000 rpm for 20 minutes at 4\u003csup\u003eo\u003c/sup\u003eC. The supernatant was transferred to a new falcon tube and again three volumes of precooled 10X TNE buffer were added. This mixture was placed in a hot water bath at 70\u003csup\u003eo\u003c/sup\u003eC for 10 minutes. After this, the mixture was cooled and again centrifuged at 6000 rpm for 20 minutes at 4\u003csup\u003eo\u003c/sup\u003eC. The resulting supernatant, which was the pure crude extract of maggots, was transferred to a new sterile falcon tube. The extract was frozen for 24 hours in a freezer, then freeze dried at -80\u003csup\u003eo\u003c/sup\u003eC. The frozen dried powder was stored at -20\u003csup\u003eo\u003c/sup\u003eC.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGC-MS analysis of crude extract\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGC-MS analysis of maggot extract was performed using a SHIMADZU GCMS QP-2010 system. An analytical column HP 5MS (30m x 0.25mm x 0.25mm) was employed, with the GC equipped with a manual splitless injector set at 240\u0026deg;C. The injector volume was 2 \u0026micro;l, and the pressure was maintained at 11.567 psi, with a split ratio of 200:1. The gas saver was turned off, and the total flow rate was set at 204 ml/min, with helium used as the carrier gas. For the oven program, the temperature was initially set at 40\u0026deg;C with a hold time of 1 minute, followed by ramping to 280\u0026deg;C at a rate of 10\u0026deg;C/min and holding for 5 minutes at this temperature. The mass spectrometer (MS) was set to a range of m/z 35-450, with a solvent delay of 0.1 minutes. The MS quad temperature was maintained at 150\u0026deg;C. The GC inlet temperature was set to 280\u0026deg;C for optimal performance. The results was observed accordingly.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIsolation of chitin from maggots\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eChitin was isolated from the larva (maggots) of house flies [4], and this process was used with some modifications and involved several steps.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDefatting\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOne hundred grams of dried maggots were collected and crushed into a fine powder using a mortar and pestle. The dried, crushed maggots were transferred to a beaker. About 500 ml (v/v) of the chloroform (Sigma-Aldrich CAS No. 67-66-3) and methanol (Sigma-Aldrich CAS No. 67-56-1) mixture (7:3) was added to the crushed, dried maggots. This mixture was placed on a hot magnet plate and stirred for about 4 hours at 30\u003csup\u003eo\u003c/sup\u003eC. After 4 hours, the mixture was allowed to cool down to room temperature and filtrated. The filtrate was washed with distilled water until a neutral pH was attained. Air-dried the defatted crude chitin in an oven at 45\u003csup\u003eo\u003c/sup\u003eC for 24 hours and measured the dry weight of the obtained material.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeproteinization\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe dried, obtained material was placed in a 5% (w/w) NaOH (Sigma-Aldrich Lot# STBH4233) solution in a ratio of 1:10. This mixture was stirred for about 2 hours at 75\u003csup\u003eo\u003c/sup\u003eC. After 2 hours, the mixture was allowed to cool down to room temperature, and then the mixture was filtrated and all the dissolved protein content was discarded. The filtrate was washed with distilled water until a neutral pH was attained. The crude chitin was air-dried at 45\u003csup\u003eo\u003c/sup\u003eC for 24 hours. The dry weight of the obtained material was measured using a weight balance.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDemineralization\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe obtained crude chitin was placed in 2% (v/v) HCl (Sigma-Aldrich CAS No. 7647-01-0) in a ratio of 1:10. This mixture was placed on a hot magnetic plate and stirred for about 2 hours at 27\u003csup\u003eo\u003c/sup\u003eC. After 2 hours, the mixture was cooled to room temperature and then it was filtrated. Filtrate was washed with distilled water until its neutral pH was maintained. The obtained material was air-dried at 45\u003csup\u003eo\u003c/sup\u003eC for 24 hours. The weight of the obtained air dried chitin was measured.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of chitosan from isolated chitin\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eChitosan was prepared from the isolated chitin by the process of deacetylation [49], with some modifications in methodology.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeacetylation of chitin\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe obtained chitin was placed in a 50% (w/w) NaOH (Sigma-Aldrich Lot# STBH4233) solution. This suspension was stirred so that chitin was completely soaked in the solution and left at room temperature for about 30 minutes. The suspension was placed on a hot magnetic plate and allowed to stir continuously at 100\u003csup\u003eo\u003c/sup\u003eC for about 2 hours. After 2 hours, the suspension was subjected to filtration and the filtrate was washed until a neutral pH was maintained. This obtained material, chitosan, was air-dried completely at 45\u003csup\u003eo\u003c/sup\u003eC for 12 hours. The obtained chitosan was measured after complete air drying.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDecolorization\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe process of decolorization was performed in order to remove excess pigments from chitosan. The obtained chitosan was placed in a 1% CH\u003csub\u003e3\u003c/sub\u003eCOOH (Sigma-Aldrich CAS No. 144-55-8) solution in a ratio of 1:100. This mixture was placed on a hot magnetic plate and allowed to continuously stir for about 30 minutes at 27\u003csup\u003eo\u003c/sup\u003eC. \u0026nbsp;The decolorized chitosan was filtrated and washed to maintain a neutral pH. Repeat the process twice for 10 minutes to remove excess pigments. The decolorized chitosan was air dried at 45\u003csup\u003eo\u003c/sup\u003eC for 12 hours. The decolorized chitosan was subjected to a ball milling machine, and a fine powder of the prepared chitosan was made at 200 rpm speed for 40-45 minutes.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFT-IR analysis of chitosan to determine degree of deacetylation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe obtained chitosan was subjected to FT-IR analysis and it was done on IRTracer-100 FTIR Spectrophotometer-Shimadzu, to determine its peaks and degree of acetylation was determined by using the formula given by [50].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of chitosan nanoparticles\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eChitosan nanoparticles were prepared using the ion-gelatin method [51], with some modifications. Different concentrations of chitosan (0.1%, 0.2%, 0.3%, 0.4%, and 0.5%) were prepared by dissolving the respective amount of chitosan in 1% (v/v) glacier acetic acid (BDH Laboratory Supplies, Lot# B261651423) at a temperature of 60\u0026deg;C on a hot magnetic plate for 30 minutes at a speed of 900 rpm. After 30 minutes, the chitosan solution was sonicated in an ultrasonic water bath for 30 minutes to remove excess bubbles and ensure a homogeneous solution. The pH of the chitosan solution was adjusted to 5 using a 20% (w/w) NaOH (Sigma-Aldrich Lot# STBH4233) solution. A 0.1% sodium tripolyphosphate (TPP) (Sigma-Aldrich CAS No. 23-85-03) solution was prepared, and its pH was also adjusted to 5 using 0.1M HCl (Sigma-Aldrich CAS No. 7647-01-0). TPP was used as a cross-linker. TPP was gradually added drop by drop into the chitosan solution at room temperature using volume ratio of 1:1 at a speed of 700 rpm on a hot magnet plate. The dripping time should be less than 5 minutes. The solution was stirred on a hot magnetic plate at 700 rpm for 30 minutes. Subsequently, the chitosan-TPP mixture was centrifuged in a refrigerated centrifuge machine (MSE Harrier 18/80) at 6000 rpm for 15 minutes at 4\u0026deg;C. The pellet was collected by discarding the supernatant.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePurification of chitosan nanoparticles\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDeionized water was added to a collected palette, and it was vortexed for a few seconds to dissolve the palette in deionized water. The palette was again refrigerated and centrifuged (MSE Harrier 18/80) at 6000 rpm at 4\u003csup\u003eo\u0026nbsp;\u003c/sup\u003eC\u0026nbsp;for 15 minutes. The supernatant was discarded, and the pure palate was collected. The collected palette was the chitosan nanoparticles.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFreeze drying of chitosan nanoparticles \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGlucose (Systerm CAS No. 50-99-7) 5% and sucrose (Sigma-Aldrich CAS No. 57-50-1) 5% were added to chitosan nanoparticles in a volume ratio of 1:1 as cryo-protectants. The nanoparticles were freeze-dried at -80\u003csup\u003eo\u0026nbsp;\u003c/sup\u003eC\u0026nbsp;for 24 hours and stored at -20\u003csup\u003eo\u003c/sup\u003eC.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCharacterization of nanoparticles\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThese chitosan nanoparticles were characterized at the Central Research Lab at Lahore College for Women University. Different characterization processes that were used to characterize nanoparticles included the following techniques:\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eX-ray diffraction (XRD)\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe obtained nanoparticles were subjected to X-ray Diffractometer in order to assess the crystalline structure of the nanoparticles. This technique directed X-rays at the sample and measured the diffraction patterns to determine the arrangement of atoms within the crystal lattice. The resulting data provided information on the crystalinity, phase composition, and other structural parameters of the nanoparticles.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eScanning electron microscopy (SEM)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSEM was employed to visualize the morphology of the nanoparticles. This method involved scanning the surface of the sample with a focused beam of electrons, which interacted with the atoms in the sample to produce images with high resolution and depth of field. SEM images allowed for the observation of the size, shape, and surface characteristics of the nanoparticles.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFourier-transform infrared spectroscopy (FTIR)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFTIR was used for identifying functional groups and the chemical composition of the nanoparticles. In this technique, the sample was exposed to infrared radiation machine (IRTracer-100 FTIR Spectrophotometer-Shimadzu), and the resulting absorption spectra were analyzed to identify specific molecular vibrations and chemical bonds present in the sample. This analysis provided insights into the chemical structure and composition of the chitosan nanoparticles.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eUV-visible spectroscopy\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUV-Vis spectroscopy was employed to analyze the optical properties of the nanoparticles. The sample was exposed to ultraviolet and visible light, and the absorption spectra were recorded to determine the electronic transitions and band gaps within the nanoparticles.\u003c/p\u003e\n\u003cp\u003eThe results of these analyses were observed to ensure the proper characterization of the chitosan nanoparticles.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAssessment of antibacterial activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBacterial strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBacterial strains were obtained from Institute of Microbiology, University of Veterinary and Animal Sciences, Lahore. These strains were\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eEscherichia coli\u0026nbsp;\u003c/em\u003e(1609, 1874, 2021)\u003c/p\u003e\n\u003cp\u003e\u003cem\u003ePseudomonas aeruginosa\u0026nbsp;\u003c/em\u003e(101, 310)\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eKlebsiella pneumoniae\u0026nbsp;\u003c/em\u003e(310)\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eStaphylococcus aureus\u0026nbsp;\u003c/em\u003e(723)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSterilization\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll glassware (including beakers, flasks, test tubes, and glass petri plates), cotton, cotton buds, Eppendorf tubes, Micropipette tips, forceps, and the inoculum loops, forceps were autoclaved at 121 \u0026deg;C and 15 psi for 15 minutes to eliminate all microorganisms and contaminants.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of media\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eVarious nutrient media were prepared to provide the necessary nutrients and growth conditions to cultivate bacteria. These media served as the basis for isolating and identifying bacteria.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNutrient agar:\u0026nbsp;\u003c/strong\u003eNutrient agar media was prepared by dissolving 14g of weighed nutrient agar (Scharlau Lot# 18155) in 500 ml of distilled water in a conical flask. The solution was thoroughly mixed in order to dissolve it, and after that, it was sterilized in an autoclave at 121\u003csup\u003eo\u003c/sup\u003eC for 15 minutes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNutrient broth:\u0026nbsp;\u003c/strong\u003eThe thirteen grams of nutrient broth (Sigma-Aldrich Lot# 8276350) powder were measured using a weight balance and added to a conical flask. It was then dissolved in 1000 ml of distilled water and gently heated to create a clear solution. The flask was covered with aluminum foil, and the media were autoclaved at 121 \u0026deg;C for 15 minutes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEosin methylene blue (EMB) agar:\u0026nbsp;\u003c/strong\u003eEMB agar (Lab M Limited Batch No. 111833/201) was prepared by measuring 37.5 grams of eosin methylene blue\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eagar and added to a conical flask. About 1000 ml of distilled water was added in the conical flask. The suspension was soaked for 10 minutes. After that, it was heated and mixed until a uniform mixture was formed. Sterilization was achieved by autoclaving at 15 psi of pressure (121 \u0026deg;C) for 15 minutes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMacConkey agar:\u0026nbsp;\u003c/strong\u003eAbout 51.6g of MacConkey agar (HIMEDIA Lot No. 0000187708) was dissolved in 1000 ml of distilled water and heated with constant stirring to completely dissolve the media. It was then autoclaved at 121\u0026deg;C temperature and 15 psi pressure for 15 minutes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBlood agar:\u0026nbsp;\u003c/strong\u003eAbout 40 g of blood agar base (Sigma-Aldrich Lot # 9874589) was measured using a weight balance. The measured blood agar base was added to a sterilized conical flask containing distilled water. The suspension was boiled until it dissolved completely. It was then autoclaved at 121\u0026deg;C for 15 minutes. After autoclaving, the mixture was cooled to 45\u0026ndash;50\u0026deg;C, and 6% sterile horse blood was aseptically added.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of media plates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe laminar flow cabinet was cleaned and disinfected with ethyl alcohol (Sigma-Aldrich CAS No. 64-17-5). UV light was turned on for 10 to 15 minutes and then switched off. Two burners were lit on both sides of the cabinet, and the blower and fluorescent light were activated. Autoclaved media were placed inside the cabinet and poured into sterilized petri plates, which were covered until they solidified. The petri plates were sealed with paraffin wax or tape to prevent contamination and placed upside down in an incubator at 37 \u0026deg;C for 24 hours.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInoculum development\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNutrient broth (3-4ml) was taken into each sterilized test tube with the help of a micropipette. Inoculated the test tubes with a pure culture of test bacterial strains and incubated at 37\u0026deg;C in a shaking incubator for 18- 24 hours.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntibacterial activity of treatments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe antibacterial activity of the maggot extract (MEx), maggot chitosan (MCs) and chitosan nanoparticles (CNPs) against different bacterial strains was determined using the disc diffusion method.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of filter paper discs\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSterile filter paper was cut into 6mm discs and soaked them in different concentrations of MEx (2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), 10 mg/ml (D)), MCs (2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), 10 mg/ml (D)), CNPs (2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), 10 mg/ml (D)), and saline water for negative control. For positive control, antibiotic susceptibility discs of imipenem (10 mcg) were used. Ciprofloxacin (5 mcg) was used as a resistant group.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisc-diffusion method\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn this method, a loop full of bacterial culture was taken from stock and inoculated in nutrient broth at 37\u0026deg;C for 24 hours. Put the broth medium on a rotatory shaker overnight at 200 rpm. After 24 hours, swab the broth media on nutrient agar using a cotton swab. \u0026nbsp;Different agar plates for each bacteria were prepared and grouped into different categories:\u0026nbsp;\u003c/p\u003e\n\u003col style=\"list-style-type: lower-roman;\"\u003e\n \u003cli\u003e\u003cstrong\u003ePositive control group\u003c/strong\u003e: exposed to standard imipenem (10 mcg);\u0026nbsp;\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eNegative control group\u003c/strong\u003e: exposed to saline water;\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eResistant groups\u003c/strong\u003e: exposed to \u0026nbsp; standard Ciprofloxacin (5 mcg);\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eTreatment group I\u003c/strong\u003e: exposed to different concentrations of maggot extract (MEx);\u0026nbsp;\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eTreatment group II\u003c/strong\u003e: exposed to different concentrations of maggot chitosan (MCs);\u0026nbsp;\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eTreatment group III\u003c/strong\u003e: exposed to different concentrations of chitosan nanoparticles (CNPs).\u0026nbsp;\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eFilter paper discs were applied to agar plates corresponding to each group. The discs were gently pressed onto the agar surface to ensure even distribution and incubated at 37 \u0026deg;C for 24 hours. After overnight incubation, zones of inhibition were measured in millimeters and compared with those of control groups.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMorphological identification of bacterial strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn the study, bacterial colonies were morphologically identified after 24 to 48 hours of incubation on different agar petri plates. The bacterial colony size, color, and shape were observed to identify the bacterial species.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMicroscopic identification of bacterial strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGram staining was used to microscopically identify bacteria into two large groups, gram-positive and gram-negative bacteria.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGram staining\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo prepare the sample, a drop of saline and a lapful of culture were mixed on a slide to form a smear by gently stirring. The smear was then fixed by passing it through the flame. Subsequently, the smear was flooded with crystal violet and left for 1 minute. Afterward, the slide was tilted slightly and rinsed with distilled water. Next, the slide was flooded gently with grams of iodine and left for 1 minute before being rinsed again with distilled water. At this stage, the smear appeared purplish in color. The slide was then decolorized by drop wise addition of 95% alcohol onto it for 5\u0026ndash;10 seconds. After rinsing with distilled water, the slide was gently flooded with the counter-stain safranin for 45 seconds and then rinsed again with distilled water. Finally, the slide was blotted dry with bibulous paper and analyzed under a light microscope at different magnification. Gram-positive bacteria stained purple, while gram-negative bacteria retained safranin and appeared pink in color\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAssessment of Anti-fungal Property\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCollection of fungal strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFungal strains were obtained from Entomology lab of the department of zoology, Lahore College For Women University, Lahore, Pakistan. The obtained fungal strains were of\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAspergillus niger\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eAspergillus flavus\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of media\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn order to cultivate fungi, SDA media were prepared to provide the necessary nutrients and growth conditions. The SDA media served as the foundation for the isolation and identification of fungi. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of Sabouraud Dextrose Agar (SDA):\u0026nbsp;\u003c/strong\u003eMeasured about 65g of SDA (Sigma-Aldrich CAS No. 57-48-7) and dissolved it in 1000ml of distilled water. The mixture was heated until completely dissolved and then autoclaved at 121\u0026deg;C and 15 psi pressure for 15 minutes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of petri plates\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter cleaning the laminar airflow chamber with ethyl alcohol (Sigma-Aldrich CAS No. 64-17-5), the UV light was turned on for 15 minutes. After 15 minutes, it was turned off, and autoclaved media were placed inside the laminar flow. The two Bunsen burners were lit, and then the media were poured into the petri plates, filling up to three-quarters of each plate. The plates were allowed to solidify in the chamber. After solidification, the plates were sealed with paraffin wax, labeled, and left to set in the incubator upside down for 24 hours at 27\u0026deg;C.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntifungal activity of treatments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe antifungal effect of maggot crude extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs) were assessed against two different molds fungus. Antifungal activity was assessed using the disc diffusion method.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreparation of filter paper discs\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSterilized filter paper was cut into 6 mm round discs and soaked in different concentrations of MEx (2.5 mg/mL (A), 5 mg/mL (B), 7.5 mg/mL (C), 10 mg/mL (D)), MCs (2.5 mg/mL (A), 5 mg/mL (B), 7.5 mg/mL (C), 10 mg/mL (D)), and CNPs (2.5 mg/mL (A), 5 mg/mL (B), 7.5 mg/mL (C), 10 mg/mL (D)). Saline water was used as a negative control and 1 mL of 2% ketoconazole (10 \u0026micro;g/mL) as a positive control. These concentrations were prepared in distilled water.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisc-diffusion method\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFrom the previous 7-day-old cultured plates of Aspergillus strains, a small amount of fungal mold was taken with the help of a sterilized swab and swabbed all around the petri plates. After swabbing, the different SDA petri plates were divided into the following groups:\u0026nbsp;\u003c/p\u003e\n\u003col style=\"list-style-type: lower-roman;\"\u003e\n \u003cli\u003e\u003cstrong\u003ePositive control group:\u003c/strong\u003e Exposed to ketoconazole (10 \u0026micro;g/mL) as a positive control and saline water as a negative control.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eTreatment group I:\u003c/strong\u003e Exposed to different concentrations of MEx.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eTreatment group II:\u003c/strong\u003e Exposed to different concentrations of MCs.\u003c/li\u003e\n \u003cli\u003e\u003cstrong\u003eTreatment group III:\u003c/strong\u003e Exposed to different concentrations of CNPs.\u0026nbsp;\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eFilter paper discs corresponding to each group were applied to the SDA plates. The discs were gently pressed onto the agar surface to ensure even distribution and incubated at 27\u0026deg;C for 5 days.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMeasurement of growth inhibition zones\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter the incubation period, the diameter of the fungal colonies on each plate was measured and recorded, and these measurements were compared to those of the control plates.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMacroscopic identification of fungi\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe morphological characters of each colony developed after 6 days. The fungal isolates were identified using standard mycological methods that involve a combination of macroscopic and microscopic examinations. The colony features (color, shape, size, and hyphae) were macroscopically observed in Petri plates to study the fungal morphology. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMicroscopic examination of fungi\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe microscopic examination was done under a microscope after staining. A glass slide with 70% alcohol and lacto phenol blue received a fungal specimen via a sterile needle. After being covered with a coverslip, microscopic features were observed under a light microscope.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data was analyzed using GraphPad Prism 8.0.2 (263), IBM SPSS Statistics 25, Microsoft Excel (2013), and Origin 2018 64 Bit. Means, standard deviations, and standard errors were calculated. Statistical tests, including ANOVA, were employed to assess the antibacterial and antifungal effectiveness of maggot extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs).\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cp\u003e\u003cstrong\u003eGC-MS analysis of crude extract\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe qualitative GC-MS analysis of the maggot crude extract revealed 20 distinct peaks; these peaks were identified by comparing their retention times (RT) with standard peaks using MS Interpreter (Ver. 3.4.5.) software. The quantitative analysis of the peaks provided the average concentration of these compounds in the extract solution (MEx). In addition to their antibacterial, antifungal, and anti-inflammatory properties, these compounds exhibit a wide array of bioactive effects suitable for potential medicinal applications. For example, several alcohols and esters found in the extract demonstrate antioxidant potential, which could be beneficial in addressing conditions related to oxidative stress. Furthermore, certain compounds, like cyclic lactones and specific fatty acid derivatives, showed promising neuroprotective properties. Table 1 illustrates all the summarized results of GC-MS of maggot crude extract compounds with their retention time, molecular weight, and formula.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eChitosan preparation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eChitosan was prepared through a series of different steps, with each step resulting in a significant reduction in material weight. It showed the degree to which each method separated and purified the chitosan from the original maggot material. The yield of each processing step are summarized in Table 2 and Figure 1 detailing the materials obtained along with the corresponding dry weights.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMeasurement of degree of deacetylation of chitosan\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe prepared chitosan underwent FT-IR analysis, and the resulting data were utilized to generate a graph spectrum between wavelengths of 4000 cm\u003csup\u003e-1\u003c/sup\u003e and 500 cm\u003csup\u003e-1\u003c/sup\u003e on the x axis and transmittance on the y axis using Origin 2018 64Bit software, as shown in Figure 2. In the FTIR (Fourier Transform Infrared) analysis of chitosan, several characteristic peaks were observed. Notable peaks included a broad absorption around 3350-3500 cm⁻\u0026sup1;, corresponding to O-H and N-H stretching vibrations, and a peak at 1650 cm⁻\u0026sup1; associated with the amide I band from C=O stretching vibrations. Additional significant peaks were observed at 1580 cm⁻\u0026sup1; for N-H bending vibrations, 1420 cm⁻\u0026sup1; for CH2 bending vibrations, and 1320 cm⁻\u0026sup1; for C-N stretching vibrations or the amide III band. Peaks at 1150 cm⁻\u0026sup1; and 1080 cm⁻\u0026sup1; represented C-O-C and C-O stretching vibrations, respectively, which were indicative of glyosidic linkages in the chitosan backbone.\u003c/p\u003e\n\u003cp\u003eThese specific peaks are important in confirming the chemical structure and composition of chitosan, as well as it provide insight into its degree of deacetylation and impurities present in the sample. According to the equations proposed by [50], the degree of deacetylation of the prepared chitosan sample was determined to be 90.57%.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePreperation of chitosan nanoparticles\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe chitosan nanoparticles were prepared using the ion-gelatin method. At concentration of 0.1% and 0.2%, no pellet formation was observed after refrigerated centrifugation. At 0.3% chitosan, a small pellet briefly formed but rapidly disintegrated, preventing its collection for further analysis. However, at a concentration of 0.4%, a stable pellet was successfully observed and collected. The palette at 0.4% chitosan maintained its structure, and this palette was used for further characterization and was used in the further study. At 0.5% chitosan concentration, chitosan was not dissolved properly in the 1% acetic acid solution, and its incomplete dissolution led to the formation of a suspension at the bottom of the flask. Due to this suspension, the cohesive pellet was formed on post-centrifugation, indicating insufficient dissolution in the solution.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCharacterization results of nanoparticles\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe characterization results of chitosan nanoparticles are discussed below.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFTIR analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe FTIR spectra of chitosan nanoparticles (CNPs) revealed significant changes induced by interactions with sodium tripolyphosphate (STPP), which were crucial for their structural and functional properties. Key observations included intensified O-H stretching vibrations at 3416 cm\u003csup\u003e-1\u003c/sup\u003e, indicating enhanced hydrogen bonding facilitated by STPP. Shifts in NH2 and OH stretching vibrations to 3292 cm\u003csup\u003e-1\u003c/sup\u003e reflected structural modifications in CSNPs compared to native chitosan. Changes in amide bands (Amide I at 1648 cm\u003csup\u003e-1\u003c/sup\u003e and Amide II at 1586 cm\u003csup\u003e-1\u003c/sup\u003e) suggested altered hydrogen bonding patterns, influenced by STPP interactions, as shown in Figure 3. Additionally, the presence of P=O stretching vibrations at 2407 cm\u003csup\u003e-1\u003c/sup\u003e confirmed the incorporation of phosphate groups from STPP, enhancing nanoparticle stability and cross-linking capabilities.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eUV-Visible analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn the UV absorbance graph of chitosan nanoparticles (CNPs), a distinct and sharp peak was observed at 247 nm, indicating the wavelength where these nanoparticles absorbed light most effectively. This peak, characterized by its straight and noise-free shape, as shown in Figure 4, represented specific electronic transitions or absorption bands inherent to the CNPs\u0026apos; molecular structure. The sharpness and clarity of the peak suggested a high degree of uniformity and consistent size distribution of the CNPs.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eXRD analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe XRD pattern of chitosan nanoparticles exhibits a prominent broad peak around 2\u0026theta; = 10\u0026deg;, indicating a significant amorphous character within the material. This broad peak is typical for polymers like chitosan, reflecting the irregular arrangement of the polymer chains. Additionally, the pattern displays several minor peaks between 20\u0026deg; and 80\u0026deg;, which suggest the presence of small crystalline regions or impurities within the predominantly amorphous matrix, as shown in Figure 5. These crystalline regions could be inherent to the semi-crystalline nature of chitosan or may have formed during the nanoparticle preparation process.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSEM analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe scanning electron microscopy (SEM) image of the chitosan nanoparticles revealed a complex morphology characterized by a mixture of amorphous and crystalline structures. At a magnification of 495x and with a scale bar of 10 \u0026micro;m, the nanoparticles showed a rough surface and had an average diameter of 262 nm. The morphology exhibited irregular, amorphous regions alongside smooth crystalline areas, indicating a heterogeneous structure, as shown in Figure 6.\u003c/p\u003e\n\u003cp\u003eThe amorphous regions of the nanoparticles display an irregular and non-uniform texture, lacking a defined shape or smoothness. In contrast, the crystalline regions exhibit smooth and well-defined surfaces, suggesting partial crystallization within the chitosan matrix.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAssessment of anti-bacterial activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntibacterial susceptibility test\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAntibacterial susceptibility tests were conducted to evaluate the effectiveness of different concentrations of maggot crude extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs) against various bacterial strains. The concentrations tested for all the above treatments included 2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), and 10 mg/ml (D). Imipenem 10 mcg (P) was used as a positive control, saline water as a negative control (N), and Ciprofloxacin (5 mcg) (R) was used as a resistance group. These treatments were applied to selected bacterial strains, and their zones of inhibition were measured to determine the antibacterial activity at each concentration. The results obtained after statistical analysis are listed below.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntibacterial activity against \u003cem\u003eE. coli\u003c/em\u003e strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAntibacterial activity of the treatments against three strains of \u003cem\u003eE. coli\u003c/em\u003e (1874, 1609, and 2021) was measured. From treatment group, MCs at concentration D (10 mg/ml) showed the highest zone of inhibition (21.6 \u0026plusmn; 0.6) against \u003cem\u003eE. coli\u003c/em\u003e 1874 and (20.3 \u0026plusmn; 0.3)\u0026nbsp;\u003cem\u003eE. coli\u003c/em\u003e 1609, as shown in Figure 7 and 8, while MEx demonstrated the highest zone of inhibition (20.3 \u0026plusmn; 0.3) against \u003cem\u003eE. coli\u003c/em\u003e 2021, as shown in Figure 9. The mean values of the zones of inhibition are listed in Table 3. Figure 10 shows the graphical representation of the zones of inhibition of the treatments against the \u003cem\u003eE. coli\u003c/em\u003e strains.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntibacterial activity against\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eP. aeruginosa\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAntibacterial activity against two strains of \u003cem\u003eP. aeruginosa\u003c/em\u003e (101 and 310) was measured. From treatment groups, MEx at concentration D (10 mg/ml) showed the highest zone of inhibition \u003cstrong\u003e(\u003c/strong\u003e29.6 \u0026plusmn; 0.6; 18.6 \u0026plusmn; 0.3)\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eagainst both \u003cem\u003eP. aeruginosa\u003c/em\u003e strains 101 and P310, as shown in Figure 11 and Figure 12. The mean values of the zones of inhibition are listed in Table 4. Figure 13 shows the graphical representation of the zones of inhibition of the treatments against the \u003cem\u003eP. aeruginosa\u003c/em\u003e strains. The results indicated that the zones of inhibition increased with the increased in the concentration of the treatments against \u003cem\u003eP. aeruginosa strains\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntibacterial activity against\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eS. aureus\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;strain\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAntibacterial activity against \u003cem\u003eS. aureus\u003c/em\u003e (723) was measured. From treatment group, MEx at concentration D (10 mg/ml) showed the highest zone of inhibition (18.6 \u0026plusmn; 0.3), as shown in Figure 14. The mean values of the zones of inhibition are listed in Table 5. Figure 15 shows the graphical representation of the zones of inhibition of the treatments against the \u003cem\u003eS. aureus\u003c/em\u003e strain. These results underscore the effectiveness of MEx at higher concentrations in inhibiting the growth of \u003cem\u003eS. aureus\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntibacterial activity against\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eK. pneumoniae\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;strain\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAntibacterial activity against \u003cem\u003eK. pneumoniae\u003c/em\u003e (K310) was measured. From treatment group, MEx at concentration D (10 mg/ml) showed the highest zone of inhibition (2.6 \u0026plusmn; 0.3), as shown in Figure 16. The mean values of the zones of inhibition are listed in Table 6. Figure 17 shows the graphical representation of the zones of inhibition of the treatments against the \u003cem\u003eK. pneumoniae\u003c/em\u003e strain. These results underscore the effectiveness of MEx at higher concentrations in inhibiting the growth of \u003cem\u003eK. pneumoniae.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis of antibacterial activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStatistical analysis of variance (ANOVA) was performed using GraphPad Prism 8.0.2 software, revealing significant variations in zones of inhibition among different treatment groups. The interaction between bacterial strains and treatments was also significant (P \u0026lt; 0.005), suggesting complex relationships influencing antibacterial efficacy across various treatments and bacterial strains. Table 7 illustrates the detailed statistical analysis of antibacterial activity.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMorphological and microscopic examination of bacterial strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of \u003cem\u003eEscherichia coli\u003c/em\u003e:\u0026nbsp;\u003c/strong\u003e \u003cem\u003eE. coli\u0026nbsp;\u003c/em\u003eis a gram-negative rod shaped and facultative bacteria. It gave pinkish red color on gram staining. It gave greenish fluorescence colonies on EMB (Eosin Methylene Blue) media and pinkish colonies on MacConkey media, as shown in Figure 18.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of \u003cem\u003eKlebsiella pneumoniae:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cem\u003eKlebsiella pneumoniae\u003c/em\u003e is a gram-negative, rod-shaped, and facultative bacterium. It produced a pinkish-red color on Gram staining. On EMB (Eosin Methylene Blue) media, it formed purple colonies usually without metallic green sheen. On MacConkey agar, it produced mucoid, pink colonies due to its ability to ferment lactose, as shown in the Figure 19.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of \u003cem\u003ePseudomonas aeruginosa:\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e is a gram-negative, rod-shaped, and obligate aerobic bacterium. It produced a pinkish-red color on Gram staining. On EMB (Eosin Methylene Blue) media, it formed colorless colonies because it does not ferment lactose. On MacConkey agar, it also produced colorless colonies, because it is a non-lactose fermenter, as shown in Figure 20.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of \u003cem\u003eStaphylococcus aureus\u003c/em\u003e:\u003c/strong\u003e\u003cem\u003e\u0026nbsp;Staphylococcus aureus\u003c/em\u003e is a gram-positive, spherical (coccus) bacterium. It gave a purple or blue appearance when observed under a microscope after Gram staining due to its thick peptidoglycan cell wall retaining the crystal violet dye. On blood agar, \u003cem\u003eStaphylococcus aureus\u003c/em\u003e produced whitish colonies, as shown in Figure 21.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAssessment of antifungal activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntifungal susceptibility test\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAntifungal susceptibility tests were conducted to evaluate the effectiveness of different concentrations of maggot crude extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs) against two different molds of Aspergillus. The concentrations tested for all the above treatments included 2.5 mg/ml (A), 5 mg/ml (B), 7.5 mg/ml (C), and 10 mg/ml (D). Ketoconazole (10 \u0026micro;g/mL) was used as a positive control (P), and normal saline water was used as a negative control (N). These treatments were applied to selected fungal molds, and their zones of inhibition were measured to determine the antifungal activity at each concentration. The results obtained after statistical analysis are described below.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAntifungal activity against Aspergillus species\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAntifungal activity against two different molds of Aspergillus (\u003cem\u003eA. flavus\u0026nbsp;\u003c/em\u003eand\u003cem\u003e\u0026nbsp;A. niger\u003c/em\u003e) was measured. From treatment groups, MEx at concentration D (10 mg/ml) showed the highest zone of inhibition ((23.3 \u0026plusmn; 0.3; 23 \u0026plusmn; 0.5)) against both molds of Aspergillus, as shown in Figure 22 and Figure 23. The mean values of the zones of inhibition are listed in Table 8. Figure 24 shows the graphical representation of the zones of inhibition of the treatments against the Aspergillus species.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis of antifungal activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data was statistically analyzed by using GraphPad Prism 8.0.2 software, ANOVA test was applied to statistically analyze the data. The results of ANOVA analysis of variance examined the zone of inhibition against fungal molds, specifically \u003cem\u003eA. flavus\u003c/em\u003e and \u003cem\u003eA. niger\u003c/em\u003e, revealed significant effects of different treatments. Significant main effects are observed for treatments (P \u0026lt; 0.005), suggesting effective relationships influencing antifungal efficacy across various treatments and fungal molds. Table 9 describes the detailed results of statistical analysis of antifungal activity.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMorphological and microscopic identification of Aspergillus species\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of \u003cem\u003eAspergillus flavus\u003c/em\u003e:\u0026nbsp;\u003c/strong\u003e\u003cem\u003eAspergillus flavus\u0026nbsp;\u003c/em\u003ecolonies, when grown on SDA (Sabouraud Dextrose Agar) media, exhibited a yellow-green surface. It showed a velvety to cotton-like texture. Under the microscope, \u003cem\u003eAspergillus flavus\u003c/em\u003e showed septate, hyaline hyphae and long, unbranched conidiophores ending in spherical vesicles covered with phialides. The rough-walled conidia were spherical, as shown in Figure 25.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of \u003cem\u003eAspergillus niger\u003c/em\u003e:\u0026nbsp;\u003c/strong\u003e\u003cem\u003eAspergillus niger\u003c/em\u003e colonies exhibited a white base with black colonies on the surface of SDA (Sabouraud Dextrose Agar) media. It showed a granular to powdery texture. Microscopic examination with lactophenol cotton blue staining revealed septate, clear hyphae and long, smooth conidiophores ending in large, round vesicles. The conidia were in large, radiating chains, forming a dense, black powdery mass, as shown in Figure 26.\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThe biomedical field has seen a growing interest in the use of natural and biocompatible materials for therapeutic applications. Among these, maggot extract, chitosan, and its derivative products have emerged as effective options. Their unique properties, including biocompatibility, biodegradability, and potent bioactivity, make them attractive for various therapeutic purposes. This study evaluated the antimicrobial activity using the disc diffusion method. All treatments demonstrated positive antimicrobial effects. However, there were certain differences in efficacy among the treatments for each group.\u003c/p\u003e \u003cp\u003eIn this study, the substantial reduction during the production of chitosan was observed which is in accordance with the previous research [\u003cspan additionalcitationids=\"CR53\" citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. The degree of deacetylation of chitosan was determined to be 90.57%, is comparable to values reported by [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e], where deacetylation levels typically range from 85\u0026ndash;95% for effective chitosan preparation. The stability of the chitosan nanoparticles pellet prepared by the ion-gelatin method at a 0.4% chitosan concentration and the sedimentation problem at higher concentrations align with findings from [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e], who reported similar challenges in nanoparticle preparation related to chitosan concentration.\u003c/p\u003e \u003cp\u003eIn the current findings, the GC-MS analysis of the maggot crude extract (MEx) identified twenty distinct compounds with diverse bioactive properties. The identified compounds include alcohols, esters, cyclic lactones, fatty acid derivatives, phthalate and n-octane, which have shown promising biological activities in previous studies. For instance, the antioxidant properties of compounds like 3-methyl-1-butanol (isoamyl alcohol) and isovaleric acid isopentyl ester are consistent with the previous findings [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e], who reported similar compounds exhibiting significant antioxidant effects. Additionally, the presence of cyclic lactones and fatty acid derivatives, including oleic acid, highlighted potential neuroprotective properties as demonstrated by previous literature [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e], who reported the neuroprotective effects of oleic acid. Diethyl phthalate stands out for its anti-inflammatory properties. Previous research [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e] also demonstrated the anti-inflammatory effects of similar compounds, including phthalates. Furthermore, the antibacterial activity of n-octane aligns with the research of [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn this research work, the characterization of the CNPs revealed key structural details. The FTIR spectra revealed intensified O-H and NH\u003csub\u003e2\u003c/sub\u003e stretching vibrations and the presence of P\u0026thinsp;=\u0026thinsp;O stretching, indicating successful cross-linking with sodium tripolyphosphate (STPP), which supports the findings of [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e], who reported that STPP enhances nanoparticle stability and cross-linking, as well as [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e], who observed similar effects of STPP on nanoparticle properties. The UV-Visible analysis of the prepared chitosan nanoparticles was identified a sharp peak at 247 nm that indicated the presence of CO group as analyzed by [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. XRD analysis of the current study revealed a broad peak around 2θ\u0026thinsp;=\u0026thinsp;10\u0026deg;, indicative of the amorphous nature of chitosan nanoparticles, with minor peaks suggesting some degree of crystalinity. This finding is consistent with previous studies by [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e], who examined an amorphous structure with minor crystalline regions in chitosan nanoparticles. SEM analysis further confirmed the heterogeneous morphology of the nanoparticles, revealing a mixture of amorphous and crystalline structures with an average particle size of 262 nm. These observations are in accordance with the findings of [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e], who reported similar structural characteristics in chitosan nanoparticles.\u003c/p\u003e \u003cp\u003eIn the present study, the results of the antibacterial susceptibility tests demonstrated notable efficacy of maggot crude extract (MEx), maggot chitosan (MCs), and chitosan nanoparticles (CNPs) against various bacterial strains. MEx exhibited the highest antibacterial activity against \u003cem\u003eE. coli\u003c/em\u003e (2021), \u003cem\u003eP. aeruginosa\u003c/em\u003e (101 and 310), \u003cem\u003eS. aureus\u003c/em\u003e (723), and \u003cem\u003eK. pneumoniae\u003c/em\u003e (310) strains, with the greatest zones of inhibition observed at the highest concentration (10 mg/ml). This finding aligns with previous studies by [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e], who demonstrated the strong antimicrobial properties of crude maggot extract. Recent literature [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e] examined the compounds within MEx and found them to be particularly effective against the tested bacterial strains. Similarly, MCs showed promising results against all tested bacterial strains, with particularly strong antibacterial effects at higher concentrations against \u003cem\u003eE. coli\u003c/em\u003e (1874 and 1609). This is consistent with the research findings of [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e], who indicated that chitosan-based materials have potent antibacterial effects due to their ability to disrupt microbial cell membranes. CNPs also exhibited antibacterial activity against all the mentioned strains, but their effectiveness was comparatively less than that of MEx and MCs. However, according to [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e], chitosan nanoparticles showed the highest antibacterial activity.\u003c/p\u003e \u003cp\u003eIn the present findings, \u003cem\u003eE. coli\u003c/em\u003e was identified by its characters like greenish fluorescence on Eosin Methylene Blue (EMB) media and pinkish colonies on MacConkey media. These observations align with the work of [\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e], who described similar characteristics for \u003cem\u003eE. coli\u003c/em\u003e. [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e] demonstrated the ability of \u003cem\u003eE. coli\u003c/em\u003e to ferment lactose, producing acid that results in color changes on these media. \u003cem\u003eKlebsiella pneumoniae\u003c/em\u003e was also confirmed by the formation of mucoid, pink colonies on MacConkey agar and purple colonies on EMB media without a metallic green sheen. These results were in accordance with previous studies by [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e], who reported similar colony morphologies and fermentation profiles for \u003cem\u003eK. pneumoniae\u003c/em\u003e. \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e, a non-lactose fermenter, formed colorless colonies on both EMB and MacConkey media, and appeared pinkish-red on Gram staining. These observations are in consistent with the findings of [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. \u003cem\u003eStaphylococcus aureus\u003c/em\u003e displayed its characteristic gram-positive, spherical morphology, with a purple or blue appearance on Gram staining due to the retention of crystal violet dye in its thick peptidoglycan cell wall. The production of whitish colonies on blood agar is indicative of its ability to grow on enriched media and form distinctive colonies. These findings are in line with the study by [\u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e73\u003c/span\u003e], which described similar staining characteristics and colony morphology for \u003cem\u003eS. aureus\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eIn the current analysis, the antifungal susceptibility tests showed significant differences in the effectiveness of various treatments against \u003cem\u003eAspergillus flavus\u003c/em\u003e and \u003cem\u003eAspergillus niger\u003c/em\u003e. Maggot crude extract (MEx) at the highest concentration (10 mg/ml) had the most notable antifungal activity, with inhibition zones of 23.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3 mm for \u003cem\u003eA. flavus\u003c/em\u003e and 23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5 mm for \u003cem\u003eA. niger\u003c/em\u003e. These results highlight the strong potential of MEx as an antifungal agent. This observation is in line with previous research by [\u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e74\u003c/span\u003e], which supports these findings. Maggot chitosan (MCs) showed impressive antifungal activity, with the highest concentration (10 mg/ml) yielding inhibition zones of 20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5 mm for both \u003cem\u003eA. flavus\u003c/em\u003e and \u003cem\u003eA. niger\u003c/em\u003e. Similarly, chitosan nanoparticles (CNPs) were effective, demonstrating inhibition zones of 22.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3 mm for \u003cem\u003eA. flavus\u003c/em\u003e and 18.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3 mm for \u003cem\u003eA. niger\u003c/em\u003e at the same concentration. These findings support the previous study by [\u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e75\u003c/span\u003e], who also found that the increased surface area and reactivity of chitosan nanoparticles enhance their effectiveness against fungal strains.\u003c/p\u003e \u003cp\u003eIn this research, \u003cem\u003eAspergillus flavus\u003c/em\u003e formed yellow-green colonies on Sabouraud Dextrose Agar (SDA) with a velvety to cotton-like texture. The observed microscopic structures were septate, hyaline hyphae, long unbranched conidiophores ending in spherical vesicles, and rough-walled spherical conidia. These findings are in line with those by [\u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e76\u003c/span\u003e], who also noted the distinctive yellow-green pigmentation and conidial structures of \u003cem\u003eA. flavus\u003c/em\u003e. On the other hand, \u003cem\u003eAspergillus niger\u003c/em\u003e produced colonies with a white base and black surface on SDA, displaying a granular to powdery texture. Microscopic examination showed septate, clear hyphae, long smooth conidiophores with large round vesicles, and dense, black conidial heads. This matches the descriptions by [\u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e77\u003c/span\u003e], who highlighted the characteristic black pigmentation and granular colony texture of \u003cem\u003eA. niger\u003c/em\u003e, along with its conidial arrangement.\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eThe findings underscore the potential of MEx, MCs, and CNPs as natural and biopolymer-based treatments as effective alternatives to traditional treatments, demonstrating significant benefit in antimicrobial effects suggest promising applications in therapeutic settings, offering new avenues for enhancing patient care and treatment outcomes. These treatments not only exhibit strong efficacy in inhibiting bacterial and fungal infections. Future research should focus on elucidating the specific mechanisms underlying their therapeutic effects, optimising their formulations for clinical use, and exploring their potential for treating a broader range of medical conditions.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eI, MAIRA MUNIR, hereby declare that this manuscript titled \u0026ldquo;\u003cstrong\u003eComparative analysis of antimicrobial properties of maggot (\u003cem\u003eMusca domestica\u003c/em\u003e) crude extract, maggot chitosan and chitosan nanoparticles\u003c/strong\u003e\u0026rdquo; is an original work, and it has not been submitted or published elsewhere. All data and findings presented in this manuscript are accurate and have been obtained through ethical research practices. I confirm that there are no conflicts of interest related to this manuscript. I have adhered to the ethical guidelines and standards for research. The study has received the necessary ethical approval from Lahore College for Women University, Lahore, Pakistan, ss per guidelines of OCCUPATIONAL AND HEALTH ACT, 2007, and follows the protocols set forth by the relevant ethical review boards.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResearch Ethical Review Committee\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eProf. Dr. Farzana Rashid\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eProf. Dr. Saima Sharif\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eDr. Shagufta Naz (Associate Professor)\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eDr. Ghazala Jabeen (Associate Professor)\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eDr. Sumera Sajjad (Associate Professor)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to Publish\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAdditionally, I confirm that all authors listed in this paper have contributed to the research, and their consent has been obtained for publication. The authors have no conflict of interest to declare.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any grant from any funding agencies.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data supporting the findings of this study are available within the paper.\u003cbr\u003e\u0026nbsp;I declare that this manuscript is my original work. The research, analysis, and conclusions presented in this manuscript are entirely my own, and no part of this work has been copied, or reproduced from any other source.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eJing YJ, Hao YJ, Qu H, Shan Y, Li DS, Du RQ. Studies on the antibacterial activities and mechanisms of chitosan obtained from cuticles of housefly larvae. 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Polymer. 2002;43(4):1541\u0026ndash;7.\u003c/li\u003e\n \u003cli\u003eQi L, Xu Z, Jiang X, Hu C, Zou X. Preparation and antibacterial activity of chitosan nanoparticles. Carbohydr Res. 2004;339(16):2693\u0026ndash;700.\u003c/li\u003e\n \u003cli\u003eChung YC, Wang HL, Chen YM, Li SL. Effect of abiotic factors on the antibacterial activity of chitosan against waterborne pathogens. Bioresour Technol. 2003;88(3):179\u0026ndash;84.\u003c/li\u003e\n \u003cli\u003eLiu XF, Guan YL, Yang DZ, Li Z, Yao KD. Antibacterial action of chitosan and carboxymethylated chitosan. Appl Polym Sci. 2001;79(7):1324\u0026ndash;35.\u003c/li\u003e\n \u003cli\u003eKim S, Fernandes MM, Matam\u0026aacute; T, Loureiro A, Gomes AC, Cavaco-Paulo A. Chitosan\u0026ndash;lignosulfonates sono-chemically prepared nanoparticles: characterization and potential applications. Colloids Surf B Biointerfaces. 2013;103:1\u0026ndash;8.\u003c/li\u003e\n \u003cli\u003eXing Y, Xu Q, Yang SX, Chen C, Tang Y, Sun S, Li X. Preservation mechanism of chitosan-based coating with cinnamon oil for fruits storage based on sensor data. Sensors. 2016;16(7):1111.\u003c/li\u003e\n \u003cli\u003eChung YC, Chen CY. Antibacterial characteristics and activity of acid-soluble chitosan. Bioresour Technol. 2008;99(8):2806\u0026ndash;14.\u003c/li\u003e\n \u003cli\u003eChung Y, Su Y, Chen C, Jia G, Wang H, Wu J. Relationship between antibacterial activity of chitosan and surface characteristics of cell wall. Acta Pharmacol Sin. 2004;25:932\u0026ndash;53.\u003c/li\u003e\n \u003cli\u003eJe J, Kim S. Chitosan derivatives killed bacteria by disrupting the outer and inner membrane. J Agric Food Chem. 2006;54(18):6629\u0026ndash;33.\u003c/li\u003e\n \u003cli\u003eYang K, Xu NS, Su WW. Co-immobilized enzymes in magnetic chitosan beads for improved hydrolysis of macromolecular substrates under a time-varying magnetic field. 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Synthesis and optimization of chitosan nanoparticles: Potential applications in nanomedicine and biomedical engineering. Caspian J Intern Med. 2014;5(3):156\u0026ndash;62.\u003c/li\u003e\n\u003c/ol\u003e\n\u003col start=\"44\"\u003e\n \u003cli\u003eShi B, Shen Z, Zhang H, Bi J, Dai S. Exploring N-imidazolyl-O-carboxymethyl chitosan for high performance gene delivery. Biomacromolecules. 2012;13(1):146\u0026ndash;53.\u003c/li\u003e\n\u003c/ol\u003e\n\u003col start=\"45\" type=\"1\"\u003e\n \u003cli\u003eFriedman AJ, Phan J, Schairer DO, Champer J, Qin M, Pirouz A, Kim J. Antimicrobial and anti-inflammatory activity of chitosan\u0026ndash;alginate nanoparticles: a targeted therapy for cutaneous pathogens. J Invest Dermatol. 2013;133(5):1231\u0026ndash;9.\u003c/li\u003e\n \u003cli\u003eHejazi R, Amiji MM. Chitosan-based gastrointestinal delivery systems. J Control Release. 2003;89(2):151\u0026ndash;65.\u003c/li\u003e\n \u003cli\u003eRosyada A, Sunarharum WB, Waziiroh E. Characterization of chitosan nanoparticles as an edible coating material. In: IOP Conference Series: Earth and Environmental Science. 2019;230(1):012043\u0026ndash;59.\u003c/li\u003e\n \u003cli\u003eSorrentino A, Gorrasi G, Vittoria V. Potential perspectives of bio-nanocomposites for food packaging applications. Trends Food Sci Technol. 2007;18:84\u0026ndash;95.\u003c/li\u003e\n \u003cli\u003eYen MT, Tseng YH, Li RC, Mau JL. Antioxidant properties of fungal chitosan from shiitake stipes. Food Sci Technol. 2007;12:255\u0026ndash;68.\u003c/li\u003e\n \u003cli\u003eKasaai MR. A review of several reported procedures to determine the degree of N-acetylation for chitin and chitosan using infrared spectroscopy. Carbohydr Polym. 2008;71(4):497\u0026ndash;508.Top of FormBottom of Form\u003c/li\u003e\n \u003cli\u003eCalvo P, Remunan-Lopez C, Vila-Jato JL, Alonso MJ. Novel hydrophilic chitosan\u0026ndash;polyethylene oxide nanoparticles as protein carriers. J Appl Polym Sci. 1997;63(1):125\u0026ndash;32.\u003c/li\u003e\n \u003cli\u003eKim MW, Han YS, Jo YH, Choi MH, Kang SH, Kim SA, Jung WJ. Extraction of chitin and chitosan from housefly, Musca domestica, pupa shells. J Entomol Res. 2016;46(5):324\u0026ndash;8.\u003c/li\u003e\n \u003cli\u003eHassan MI, Mohamed AF, Taher FA, Kamel MR. Antimicrobial activities of chitosan nanoparticles prepared from Lucilia cuprina maggots (Diptera: Calliphoridae). J Egypt Soc Parasitol. 2016;46(3):519\u0026ndash;26.\u003c/li\u003e\n \u003cli\u003eHasaballah AI. Crude and chitosan nano-particles extracts of some maggots as antioxidant and anticancer agents. J Adv Biol Biotechnol. 2019;21(1):1\u0026ndash;8.\u003c/li\u003e\n \u003cli\u003eBangun H, Tandiono S, Arianto A. Preparation and evaluation of chitosan-tripolyphosphate nanoparticles suspension as an antibacterial agent. J Appl Pharm Sci. 2018;8(12):147\u0026ndash;56.\u003c/li\u003e\n \u003cli\u003eKaskoniene V, Ruockuviene G, Kaskonas P, Akuneca I, Maruska A. Chemometric analysis of bee pollen based on volatile and phenolic compound compositions and antioxidant properties. J Food Anal Methods. 2015;8:1150\u0026ndash;63.\u003c/li\u003e\n \u003cli\u003eSong J, Kim YS, Lee DH, Lee SH, Park HJ, Lee D, Kim H. Neuroprotective effects of oleic acid in rodent models of cerebral ischaemia. Sci Rep. 2019;9(1):10732\u0026ndash;41.\u003c/li\u003e\n \u003cli\u003eMahmood F, Khan JA, Mahnashi MH, Jan MS, Javed MA, Rashid U, Bungau S. Anti-inflammatory, analgesic and antioxidant potential of new (2S, 3S)-2-(4-isopropylbenzyl)-2-methyl-4-nitro-3-phenylbutanals and their corresponding carboxylic acids through in vitro, in silico and in vivo studies. Molecules. 2022;27(13):4068\u0026ndash;73.\u003c/li\u003e\n \u003cli\u003eYarinich LA, Burakova EA, Zakharov BA, Boldyreva EV, Babkina IN, Tikunova NV, Silnikov VN. Synthesis and structure\u0026ndash;activity relationship of novel 1,4-diazabicyclo[2.2.2]octane derivatives as potent antimicrobial agents. Eur J Med Chem. 2015;95:563\u0026ndash;73.\u003c/li\u003e\n \u003cli\u003eVaezifar S, Razavi S, Golozar MA, Karbasi S, Morshed M, Kamali M. Effects of some parameters on particle size distribution of chitosan nanoparticles prepared by ionic gelation method. J Clust Sci. 2013;24:891\u0026ndash;903.\u003c/li\u003e\n \u003cli\u003eAnand M, Sathyapriya P, Maruthupandy M, Beevi AH. Synthesis of chitosan nanoparticles by TPP and their potential mosquito larvicidal application. Front Lab Med. 2018;2(2):72\u0026ndash;8.\u003c/li\u003e\n \u003cli\u003eKarimi MH, Mahdavinia GR, Massoumi B. pH-controlled sunitinib anticancer release from magnetic chitosan nanoparticles crosslinked with \u0026kappa;-carrageenan. Mater Sci Eng C. 2018;91:705\u0026ndash;14.\u003c/li\u003e\n \u003cli\u003eAlehosseini E, Shahiri Tabarestani H, Kharazmi MS, Jafari SM. Physicochemical, thermal, and morphological properties of chitosan nanoparticles produced by ionic gelation. Foods. 2022;11(23):3841\u0026ndash;56.\u003c/li\u003e\n \u003cli\u003eZheng C, Zhou L. Antibacterial potency of housefly larvae extract from sewage sludge through bioconversion. Environ Sci. 2013;25(9):1897\u0026ndash;905.\u003c/li\u003e\n \u003cli\u003eAmer MS, Hammad KM, Hasaballah AI, Shehata AZI, Saeed SM. Effectiveness evaluation of Chrysomya albiceps (Diptera: Calliphoridae) and Musca domestica (Diptera: Muscidae) maggots extracts as antimicrobial and antiviral agent. Egypt J Aquat Biol Fish. 2019;23(3):561\u0026ndash;73.\u003c/li\u003e\n \u003cli\u003eRashid S, Momeni-Moghaddam M, Ghavidel Z. Lucilia sericata maggot extract: a promising tool against biofilms of antimicrobial resistant strains of Staphylococcus aureus and Pseudomonas aeruginosa. Med Bacteriol. 2024;12(1):43\u0026ndash;58.\u003c/li\u003e\n \u003cli\u003eWang L, Pang Y, Xin M, Li M, Shi L, Mao Y. Evaluating the antibacterial and antibiofilm activities of chitosan derivatives containing six-membered heterocyclics against E. coli and S. aureus. Colloids Surf B Biointerfaces. 2024;242:114084\u0026ndash;97.\u003c/li\u003e\n \u003cli\u003eObuotor TM, Ejalonibu MA, Kolawole AO, Sanusi JF, Alake AO. Comparative study of antimicrobial activities of chitosan, chitosan-silver nanocomposite, and chitosan-ampicillin hybrid on selected bacteria. Sustain Technol. 2024;13(1):1\u0026ndash;11.\u003c/li\u003e\n \u003cli\u003eRidah BA, Minai-Tehrani D, Hamad AS. Isolation and identification of Escherichia coli from children with urinary tract infection in Kirkuk city. Cent Asian J Med Nat Sci. 2024;5(1):1\u0026ndash;7.\u003c/li\u003e\n \u003cli\u003eBakshi P, Bhowmik A, Ahsan S, Alim SR. Identification of antibiotic-resistant pathogens and virulence genes in Escherichia coli isolates from food samples in the Dhaka University campus of Bangladesh. Food Sci Nutr. 2024;12(3):1995\u0026ndash;2002.\u003c/li\u003e\n \u003cli\u003eAlaa OA, Elzeftawy HM, Salama HF. Advanced studies on extended spectrum beta-lactamase producing Enterobacteriaceae in dairy cattle farms at Behaira province. Adv Vet Res. 2024;14(3):470\u0026ndash;4.\u003c/li\u003e\n\u003c/ol\u003e\n\u003col start=\"72\"\u003e\n \u003cli\u003eAkrami S, Ekrami A, Jahangirimehr F, Yousefi Avarvand A. High prevalence of multidrug-resistant Pseudomonas aeruginosa carrying integron and exoA, exoS, and exoU genes isolated from burn patients in Ahvaz, southwest Iran: A retrospective study. Health Sci Rep. 2024;7(6):2164\u0026ndash;78.\u003c/li\u003e\n \u003cli\u003eMiguel CF, Pereira CC. Evaluation of the feasibility for replacing sheep blood with human blood in culture media used in microbiological diagnostics. Anais Acad Bras Cienc. 2024;96(2):20231168\u0026ndash;89.\u003c/li\u003e\n \u003cli\u003eEl-Bassiony GM, Stoffolano JJG. In-vitro antimicrobial activity of maggot excretions/secretions of Sarcophaga (Liopygia) argyrostoma (Robineau-Desvoidy). Afr J Microbiol Res. 2016;10(27):1036\u0026ndash;43.\u003c/li\u003e\n \u003cli\u003ePoznanski P, Hameed A, Orczyk W. Chitosan and chitosan nanoparticles: parameters enhancing antifungal activity. Molecules. 2023;28(7):2996\u0026ndash;3007.\u003c/li\u003e\n \u003cli\u003eJeawel FB, Tawoos TAWAA, Ibrahim RA, Nayef AM, Hassan AA, Jaber DT, Muhaisen NA. Estimate correlation between Aspergillus spp. and asthma patients. Curr Clin Med Educ. 2024;2(6):36\u0026ndash;47.\u003c/li\u003e\n \u003cli\u003ePal M. Isolation and identification of Aspergillus niger from the onions in Bharuch, Gujarat, India. Adv Microbiol Res. 2024;5(1):178\u0026ndash;80.Top of Form\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1: Summarized results of GC-MS analysis of maggot crude extract (MEx).\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSr.No.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCompound Name\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eArea\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eArea%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMolecular Weight\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMolecular Formula\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e2.409\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e3-methyl-1-butanol (isoamyl alcohol)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e1285204\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e30.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e12\u003c/sub\u003eO\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e2.975\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003en-Octane\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e71570\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e1.71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e114\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003en-C\u003csub\u003e8\u003c/sub\u003eH\u003csub\u003e18\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e6.431\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e1-(5-Bicyclo [2.2.1] heptyl) ethylamine\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e51030\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e1.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e139\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e9\u003c/sub\u003eH\u003csub\u003e17\u003c/sub\u003eN\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e7.541\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003en-Pentanal or Valeraldehyde\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e31265\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e7.646\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eIsovaleric acid, Isopentyl ester.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e300577\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e7.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e172\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e10\u003c/sub\u003eH\u003csub\u003e20\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e12.019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eFurfuryl alcohol, tetrahydro-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e88655\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e2.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e102\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e10\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e14.765\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eDiethyl phthalate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e494809\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e11.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e222\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e12\u003c/sub\u003eH\u003csub\u003e14\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e18.542\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eZ,Z-8,10-hexadecadien-1-ol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e49884\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e1.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e238\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e16\u003c/sub\u003eH\u003csub\u003e30\u003c/sub\u003eO\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e18.948\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e9-Octadecenoic acid (Z)-, methyl ester\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e40653\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e0.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e296\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e19\u003c/sub\u003eH\u003csub\u003e36\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e20.286\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eOleic acid (9Z)-, 9-octadecenoic acid (Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e294803\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e7.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e282\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e20.346\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eDimethoxybicyclo [3.3.1]nona-2,4-dione\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e460266\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e212\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e11\u003c/sub\u003eH\u003csub\u003e16\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e22.249\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e9,12-Octadecadienoic acid (Z,Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e45124\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e1.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e280\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e32\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e22.297\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eZ,Z-3,13-Octadecadien-1-ol (4Z,13Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e29522\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e0.71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e266\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e23.429\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eZ,Z-2,13-Octadecadien-1-ol (2Z,13Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e25609\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e0.61\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e266\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e23.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eZ,Z-3,13-Octadecadien-1-ol (4Z,13Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e84265\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e2.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e266\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e23.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eZ,Z-2,13-Octadecadien-1-ol (2Z,13Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e89464\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e266\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e24.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eZ,Z-2,13-Octadecadien-1-ol (2Z,13Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e493430\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e11.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e266\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e24.265\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e6-Octadecenoic acid, methyl ester (Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e193460\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e4.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e296\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e19\u003c/sub\u003eH\u003csub\u003e36\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e24.527\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e6-Octadecenoic acid, methyl ester (Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e70025\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e1.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e296\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e19\u003c/sub\u003eH\u003csub\u003e36\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e25.755\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003eOleic acid, 9-octadecenoic acid (Z)-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 10px;\"\u003e\n \u003cp\u003e44280\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e1.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e282\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003eC\u003csub\u003e18\u003c/sub\u003eH\u003csub\u003e34\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2: Summarized results of chitosan prepared from \u003cem\u003eMusca domestica.\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 26px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eProcess\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMaterial obtained\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDry weight (grams)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 26px;\"\u003e\n \u003cp\u003eDefatting\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003eDefatted crude chitin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e88.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 26px;\"\u003e\n \u003cp\u003eDeproteinization\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003eDeproteinized crude chitin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e9.36\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 26px;\"\u003e\n \u003cp\u003eDemineralization\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003eChitin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e5.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 26px;\"\u003e\n \u003cp\u003eDeacetylation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003eChitosan\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e3.10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 26px;\"\u003e\n \u003cp\u003eDecolorization\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 40px;\"\u003e\n \u003cp\u003eDecolorized Chitosan\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 32px;\"\u003e\n \u003cp\u003e2.55\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3: Antibacterial activity of treatments against \u003cem\u003eE. coli\u0026nbsp;\u003c/em\u003estrains.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"17\" style=\"width: 11px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" style=\"width: 88px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZones of inhibition in mm (mean\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026plusmn; SE) against \u003cem\u003eE. coli\u0026nbsp;\u003c/em\u003estrains\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 36px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 21px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eE. coli\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e(1874)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eE. coli\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e(1609)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eE. coli\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e(2021)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" style=\"width: 15px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eControl\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 21px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 21px;\"\u003e\n \u003cp\u003e23.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e42.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e41 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 21px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNegative Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 21px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 21px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eResistance\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 21px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"12\" style=\"width: 15px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\" style=\"width: 21px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MEx\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e11 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e12.6 \u0026plusmn; 0.3\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e15.6 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e13.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e14.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e19.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e16.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e15.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e18.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e14.6 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e17.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e20.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 21px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e11.6 \u0026plusmn;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e14 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e13 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e12.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e18 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e12 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e14.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e16 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e14 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e21.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e20.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e18 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 21px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of CNPs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e8.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e11.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e7.3\u0026plusmn;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e9 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e8.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e9.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e9.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e9.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e12.6 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 6px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e9.5 \u0026plusmn; 0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e12 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e13.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4: Antibacterial activity of treatments against\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eP. aeruginosa\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;strains.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"17\" style=\"width: 12px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"5\" style=\"width: 87px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZones of inhibition in mm (mean\u0026nbsp;\u003c/strong\u003e\u0026plusmn;\u003cstrong\u003e\u0026nbsp;SE) against\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eP. aeruginosa\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;strains\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 16px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eControl Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 26px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 22px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eP. aeruginosa\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e(101)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 22px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eP. aeruginosa\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e(310)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 26px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e44.3\u0026nbsp;\u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e37.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 26px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNegative Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 26px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eResistance\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"12\" style=\"width: 16px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\" style=\"width: 22px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MEx\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e12.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e9.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e18.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e10.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e23.3 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e12.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e29.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e18.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 22px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e15.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e10.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e13 \u0026plusmn; 1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e11 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e15.6 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e13 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e17.6 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e17 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 22px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of CNPs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e11 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e10 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e16 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e11.14 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e11.67 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e11.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 4px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e12.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 22px;\"\u003e\n \u003cp\u003e12.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5: Antibacterial activity of treatments against\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eS. aureus\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"17\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" style=\"width: 72px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZones of inhibition in mm (mean\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026plusmn; SE) against \u003cem\u003eS. aureus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 27px;\"\u003e\n \u003cp\u003eControl Groups\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 28px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 16px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eS. aureus\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;(723)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 28px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e42.3\u0026nbsp;\u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 28px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNegative Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 28px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eResistance\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"12\" style=\"width: 27px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\" style=\"width: 20px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MEx\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e11 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e14 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e18.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e18.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 20px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e10 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e8.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e13\u0026nbsp;\u0026plusmn;\u0026nbsp;0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e17.3 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 20px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of CNPs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e8.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e11.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e11 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" style=\"width: 16px;\"\u003e\n \u003cp\u003e9.3 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 6: Antibacterial activity of treatments against\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eK. pneumoniae\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"17\" style=\"width: 13px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" style=\"width: 86px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZones of inhibition in mm (mean\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026plusmn; SE)\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eagainst\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eK. pneumoniae\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 17px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eControl Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 31px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eK. pneumoniae\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e(310)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 31px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e41.6\u0026nbsp;\u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 31px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNegative Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 31px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eResistance\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"12\" style=\"width: 17px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\" style=\"width: 24px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MEx\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e11.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e16.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e18.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e22.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 24px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e12.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e15.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e13 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e17.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 24px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of CNPs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e9.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e8.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e11.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 7px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 36px;\"\u003e\n \u003cp\u003e12.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 7:\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eStatistical analysis of antibacterial activity.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eANOVA test\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSum of square (SS)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 22px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDegree of freedom (DF)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 20px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean square (MS)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRow Factor\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17px;\"\u003e\n \u003cp\u003e7994\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 22px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 20px;\"\u003e\n \u003cp\u003e571\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e82.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003eP \u0026lt; 0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eColumn Factor\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17px;\"\u003e\n \u003cp\u003e190.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 22px;\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 20px;\"\u003e\n \u003cp\u003e31.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\n \u003cp\u003e4.601\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003e\u0026nbsp;P \u0026lt; 0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eResidual\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17px;\"\u003e\n \u003cp\u003e580.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 22px;\"\u003e\n \u003cp\u003e84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 20px;\"\u003e\n \u003cp\u003e6.907\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 9px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 8: Antifungal activity of treatments against\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAspergillus species.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"16\" style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"5\" style=\"width: 86px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eZones of inhibition in mm (mean\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e\u0026plusmn; SE) against Aspergillus molds\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" style=\"width: 18px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eControl Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 30px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eA. flavus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003eA. niger\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePositive Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e29.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e35.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 30px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eNegative Control\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"12\" style=\"width: 18px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTreatment Groups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"4\" style=\"width: 25px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MEx\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e15 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e14 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e17.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e15 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e18 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e17.3 \u0026plusmn; 0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e23.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e23 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 25px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of MCs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e15 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e15 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e16 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e15.6 \u0026plusmn; 0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e18 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e17 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e20 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e20 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"4\" style=\"width: 25px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eConcentrations of CNPs\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e13 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e11.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e15.3 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e15 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e16 \u0026plusmn; 0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e16.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 5px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eD\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e22.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 18px;\"\u003e\n \u003cp\u003e18.6 \u0026plusmn; 0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 9: Statistical analysis of antifungal activity.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eANOVA test\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSum of square (SS)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDegree of freedom (DF)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean square (MS)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eRow Factor\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003e1788\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e127.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17px;\"\u003e\n \u003cp\u003e61.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003eP\u0026lt;0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eColumn Factor\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003e0.972\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e0.972\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17px;\"\u003e\n \u003cp\u003e0.4699\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003eP\u0026lt;0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 14px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eResidual\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 15px;\"\u003e\n \u003cp\u003e28.96\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e2.068\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 17px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width: 14px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n"}],"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":"alternative therapeutics, biomedical applications, pathogen inhibition, bioactive compounds, natural antimicrobial agents","lastPublishedDoi":"10.21203/rs.3.rs-6084200/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6084200/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eMaggot metabolites exhibit strong antibacterial and pro-inflammatory properties, making them a significant focus of scientific research.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAim: \u003c/strong\u003eThis study aimed to compare the antimicrobial properties of maggot extracts (MEx), maggot chitosan (MCs), and maggot chitosan nanoparticles (CNPs).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethodology: \u003c/strong\u003eMaggot chitosan with 90.57% degree of deacetylation was extracted. CNPs were synthesized via ionotropic gelation with sodium tripolyphosphate and the characterization results confirmed their semi-crystalline structure with particle size of 262 nm.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eGC-MS analysis of MEx identified twenty bioactive compounds in MEx. By using disc diffusion method, all treatments showed significant antibacterial activity (\u003cem\u003eP = 0.0004\u003c/em\u003e) against different bacterial strains. MEx exhibited the highest antibacterial activity against \u003cem\u003eE. coli\u003c/em\u003e (2021), \u003cem\u003eP. aeruginosa\u003c/em\u003e (101 and 310), \u003cem\u003eS. aureus\u003c/em\u003e (723) and \u003cem\u003eK. pneumoniae\u003c/em\u003e (310) with a zones of inhibition 20.3 ± 0.3 mm, 29.6 ± 0.6 mm, 18.6 ± 0.3 mm, 18.6 ± 0.3 mm and 22.6 ± 0.3 mm respectively. MCs showed the highest antibacterial activity against \u003cem\u003eE. coli\u003c/em\u003e strains (1876 and 1609) with zones of inhibition 31.6 ± 0.6 mm and 20.3 ± 0 mm. The antibacterial activity of CNPs was comparatively lower than that of MEx and MCs. In antifungal susceptibility tests, all treatments were significantly sensitive to both Aspergillus molds (\u003cem\u003eP = 0.0001\u003c/em\u003e). However, MEx showed the highest antifungal activity against \u003cem\u003eA. flavus\u003c/em\u003e and \u003cem\u003eA. niger\u003c/em\u003ewith zones of inhibition 23.3 ± 0.3 mm and 23 ± 0.5 mm.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFindings:\u003c/strong\u003e The findings suggest that these cost-effective medications would be very effective in preventing infections in the body. Additionally, they have minimal side effects. Further research and development could lead to new, effective medications based on these agents.\u003c/p\u003e","manuscriptTitle":"Comparative analysis of antimicrobial properties of maggot (Musca domestica) crude extract, maggot chitosan and chitosan nanoparticles","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-03-06 08:33:40","doi":"10.21203/rs.3.rs-6084200/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":"22585afa-daf7-405d-9a72-d4d86555a3f2","owner":[],"postedDate":"March 6th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":45210641,"name":"Biological sciences/Biochemistry"},{"id":45210642,"name":"Biological sciences/Microbiology"},{"id":45210643,"name":"Biological sciences/Zoology"},{"id":45210644,"name":"Health sciences/Diseases"},{"id":45210645,"name":"Physical sciences/Nanoscience and technology"}],"tags":[],"updatedAt":"2025-03-31T06:09:21+00:00","versionOfRecord":[],"versionCreatedAt":"2025-03-06 08:33:40","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6084200","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6084200","identity":"rs-6084200","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}