Introduction
In comparison to traditional physical and chemical approaches, the development of natural resources for human health through the synthesis of nanoparticles (NPs) utilizing materials from animals (peels, fish, blood, bones, etc.) results in advancements in the production of low-cost, environmentally friendly, and long-lasting stabilized NPs. Massive population growth and a quick biological revolution have become significant environmental contamination characteristics. When used in medication, pesticides and insecticides might readily float into water or be transported by the wind, producing several hazardous substances that could harm human health and the environment (Luceri, Francese, Lembo, Ferraris, & Balagna, 2023; Mercan, Niculescu, & Grumezescu, 2022; Sharmin et al., 2021). On the other hand, pathogenic microbes participate in releasing highly toxic pollutants, such as Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, and Candida. As a result, early monitoring and detection of these dangerous bacteria has emerged as a critical strategy for success. Nanoparticles are an essential component of this technology that provides workable solutions (Khan, Hamadneh, & Khan, 2016). Three categories comprise the different types of nanoparticles: metal oxide, nanocomposite, and nano-metal (Khan, Saeed, & Khan, 2019). Although there are several applications for nanomaterials, little study has been done on nanocomposite, particularly concerning harmful bacteria (Prabhu & Poulose, 2012). Both positive and negative bacteria severely threaten human health and the environment. When extreme toxicity is introduced into human systems in large concentrations, it can be fatal. It is essential to develop nanoparticles that can rapidly eradicate this bacterium for human health and environmental preservation (Prabhu & Poulose, 2012). The toxic substances secreted by bacteria and fungi are endotoxins and aflatoxin, and their pollutants affect the airways, mouth, digestive, reproductive, and urinary tracts, as well as the contamination of food and feed with mycotoxins.
In the last few decades, metal oxides like AgO, SnO2, ZnO, Fe2O3, CuO, and beyond (Galúcio et al., 2022). Marine shrimp is a popular marine variety and a good source of protein. While marine shrimp contains few calories, it contains many essential nutrients. These nutrients include selenium, a mineral that shields cells from damage, and vitamin B12, which helps create red blood cells and maintains the neurological system's normal operation (USDA, 2020; USEPA, ). Because of their intriguing physicochemical properties, iron oxide nanoparticles (IO-NPs) have also gained traction in research areas looking into anticancer, antifungal, plant growth, and antibacterial applications (Yamada, Seno, Masuoka, & Yamashita, 1998). Because plant-based nanoparticles are more stable and economical than those derived from microbes, their biological production has potential value (Betty, Choudhury, & Shah, 2022). Because of its regulated size, strong magnetic, and low toxicity, IO-NPs' nanoparticle activity is quite significant (Nasiri, 2023). To the best of the author's knowledge, the reason for this sort of work was the shortage of reports and studies about the use of PLA (Nd-YAG) procedures during the research on animal extracts. The primary objective of this work is to compare the AgO/Fe2O3-NCs utilizing the PLA (Nd-YAG) approach. The PLA (Nd-YAG) process can lead to higher purity, a suitable crystalline structure, non-toxicity, safety, affordability, and environmental friendliness.
Marine shrimp are low in fat and abundant in minerals and proteins, making them beneficial for health. Furthermore, two essential biological properties of thin products are the chemopreventive and chemoprotective properties of its lipids. Several biological activities of methanolic and lipidic extracts of shrimp waste and muscle have been reported. Through techniques gathered under the term "chemoprevention," which refers to using natural or synthetic compounds to prevent cancer and microbial formation, these actions, which can influence biological processes, have been related to cancer prevention (López-Saiz, Suárez-Jiménez, Plascencia-Jatomea, & Burgos-Hernández, 2013). One kind of shrimp that works well with particle structure is the white shrimp.
The previous studies in 2024 on iron oxide NP synthesis via a chemical method on bacterial cells (gram-positive and gram-negative) by Rishikesh Kumar et al. (Kumar, Kumari, & Sahoo, 2024). In 2024, metal and metal oxide nanoparticles will be synthesized using green synthesis for medical applications. Study effect size, shape, and distribution of NPs in their toxicity by de Jesus, R. A., et al. (Jesus et al., 2024).
In this work, the AgO/Fe2O3-NCs preparation involves mixing the marine shrimp shell extract with Ag2NO3 and FeCl3 salt for the first time via a PLA (Nd-YAG) method at 300 oC for 2 hours for antimicrobial activity. AgO/Fe2O3-NCs were characterized using XRD analysis (XRD-6000/Japan), and the JCPDS card (Joint Committee on Powder Diffraction Standards) was used to compare the findings. XRD measurements in the Nanotechnology Laboratory and Advanced Materials/The Materials Research Department/The Ministry of Science and Technology in Iraq have been used to examine the orientation of AgO/Fe2O3-NCs developed samples. AgO/Fe2O3-NCs shape and particle size were analyzed using FE-SEM, TEM, and EDS-Mapping, tested in Iran-Mashhad at "Tescan Mira3 SEM-Czechia.". The absorption peaks, the compound, and the functional group were determined via the FTIR spectrum (Perkin-Elmer Spectrum GX FTIR).
Experimental part
Preparation of the marine shrimp shell extract
The biological name of the marine shrimp shell is Caridean (Islam et al., 2024). The local market in Baghdad, Iraq, provided the shells of marine shrimp. Marine shrimp shell samples taken for this investigation were taken to remove contaminants, sliced into little pieces, and then dried for nine days in an Umbra case. The dehydrated samples were then ground into a fine powder in a professional blender made of stainless steel. 200 ml of deionized water and 10 g of powdered mixture were used to create animal extracts. A magnetic stirrer brought The solution to a boil at 80 °C for two hours. The solution was frozen at room temperature and filtered through a Whatman filter sheet. Figure 1 (A–C) shows the stages of making the marine shrimp shell extract.
Preparation of AgO/Fe2O3 NCs from marine shrimp shell extract by PLA (Nd-YAG technique)
To prepare AgO/Fe2O3-NCs, a magnetic stirrer was used to dissolve 5 g of Ag2NO3 in 100 ml of deionized water for 60 minutes at 5000 rpm/min. Next, following our earlier research, a FeCl3 solution was added to 100 millilitres of deionized water and exposed to PLA (Nd: YAG) radiation (Hajiali, Daneshjou, Daneshjoo, & Khajeh, 2024). After that, 100 ml of marine shrimp extract was mixed with 200 ml of Ag2NO3 and Fe2O3 solutions. This colloid was irradiated with 200 Mj and 1500 pulses at 6 Hz. The distance between the laser gun and the sample is 5 cm. The AgO/Fe2O3 NCs colloidal solution was created in the same solution and then exposed to another round of laser radiation. For seventy minutes, the two solutions were combined using a magnetic stirrer. Deionized water was used to wash the precipitate many times. For three hours, the drying procedure was conducted in an oven set at 200°C. Experimental conditions such as laser parameters, reaction environment, and precursor concentrations are shown in Table 2 .
Characterization of AgO/Fe2O3 NCs synthesized by the PLA (Nd-YAG technique)
The Shimadzu-Japan XRD-6000 apparatus was utilized, and its measurement range was 10 to 80°. At 40 kV and 30 mA, the X-ray generator ran. The XRD instrument's scan range was used to compare the acquired diffraction patterns to the information supplied by the Joint Committee on Powder Diffraction Standards (JCPDS) card. The function groups are identified using the FTIR spectrum (Perkin-Elmer Spectrum GX FTIR). FTIR spectroscopy is An essential tool for determining a substance's molecular structure. A molecule's basic parts and bonding arrangement can be ascertained by examining the single resonant vibrational modes of its different branches or body regions. Since each molecule has a distinct infrared spectrograph that may be used to identify it, the opposite process is also employed, much like a fingerprint. Over the past ten years, FTIR spectroscopy analysis—a technique based on the concept of infrared spectroscopy—has found more use in researching materials at the nanoscale (Baudot, Tan, & Kong, 2010). Using a FESEM device, Tuscan Mira3 FESEM-Czechia in Mashhad, Iran, has evaluated the samples. One of the most commonly used methods for characterizing nanoparticles is FE-SEM with Energy Dispersive Spectrometry (EDS).
Because of their small size (1–100 nm), optical microscopes cannot detect them, necessitating such approaches. High-resolution imaging in FE-SEM enables the imaging of particles, while EDS provides elemental and chemical information on nanoparticles. (FE-SEM and TEM) offer essential insights into the size and shape of the generated nanoparticles (NPs). At various magnifications, the FE-SEM and TEM fields yielded microstructural and nanoscale data, such as particle size and morphological features, at a voltage of 20.00 KV. TEM was the most advanced and effective technology for imaging materials. However, the TEM tools could not handle the demands of photographing such one-nm-sized atom clusters due to insufficient spatial resolution. Feynman emphasized the need for improved spatial resolution in TEM equipment (Anjum, 2016).
Antimicrobial activity of AgO/Fe2O3 NCs prepared from marine shrimp shell extract by PLA (Nd-YAG) technique
The antimicrobial potential of the prepared samples (AgO/Fe2O3-NCs) was investigated against gram-negative and gram-positive bacterial strains using an agar-well diffusion assay. About 20 mL of Muller-Hinton (MH) agar was aseptically poured into sterile Petri dishes via diffusion. The bacterial species were collected using a sterile wire loop from their stock cultures. After culturing the organisms, six mm-diameter wells were bored on the agar plates using a sterile tip. In the bored wells, 30 mg/L concentrations of the samples (AgO/Fe2O3-NCs) worked. After being incubated at 37°C for the whole night, the test organisms and the AgO/Fe2O3-NCs samples were placed on cultivated plates. The average diameter of the zones of inhibition was then measured and recorded. The University of Baghdad—College of Education for Pure Science (Ibn al-Haitham) microbiology lab supplied the microbial cultures. This test was conducted using sterile petri plates with a 90-mm diameter that contained sterile nutritional agar medium using the agar-well diffusion technique. In 1000 millilitres of deionized water, 0.5 grams of peptone—which supplies organic nitrogen—0.3 grams of yeast extract—which supplies vitamins, carbohydrates, nitrogen, and salts—and 28 grams of agar—which gives the combination solidity—were dissolved to create nutrient agar medium. At 27 oC, the pH was corrected to 6.9. After mixing these components and boiling them for around 15 minutes at 121 oC to ensure they are well incorporated, the mixture is placed into petri dishes and covered immediately to solidify. Next, AgO/Fe2O3-NCs were dissolved in a 30 mg/mL solution of dimethyl sulfate (DMSO) solvent. The Petri plate surface was swabbed with the recently created microorganisms. Using a sterile gel puncher or cork borer, 8 mm diameter wells were made in the medium containing bacterial and fungal life for every plate. For bacteria and fungi, 40 μL of AgO/Fe2O3-NCs and DMSO control are present in each well at the appropriate concentration. The test specimens were incubated for 24 hours at ±25 oC to grow bacteria and fungus. The width of the inhibitory zone was then measured in millimetres. A bacterial culture dish was fertilized for AgO/Fe2O3-NCs by the PLA (Nd-YAG) technique, explaining the different levels of inhibition of zones in the dose-depending style. The AgO/Fe2O3-NCs intrinsically exhibit high levels of inhibition of zone versus gram-negative (Escherichia coli) and gram-positive (Staphylococcus aureus, Staphylococcus epidermidis). The following calculation has been used to calculate the percentage of the inhibition zone (Hajiali et al., 2024):
Inhibition zone(%)=
Result and Discussion
XRD of AgO/Fe2O3 NCs prepared from marine shrimp shell extract by PLA (Nd-YAG technique)
The XRD pattern was used for the structure-properties study of AgO/Fe2O3 NCs prepared from the marine shrimp shell extract via the PLA (Nd-YAG) technique at 200 Mj with 1500 pulses. Figure 3 and Table 3 show good crystallinity and many diffraction peaks for AgO/Fe2O3 NCs. The crystal structure of AgO/Fe2O3 NCs is cubic-like and has been shown and matched with (00-087-1526). There are four more distinguished peaks below the AgO NCs at 32.1o, 46.6o, 67o, and 76.1o that correspond to (111), (200), (220), and (311), respectively. Fe2O3 nanoparticles produce overlapping diffraction angles with miller (012), (024), and (116) patterns. The crystallite size of AgO/Fe2O3 NCs is 9.6 to 19.5 nm. The formula developed by Debye Scherer was used to determine the crystallite size of AgO/Fe2O3 NCs (Abid & Kadhim, 2020; Abid, Abid, Aziz, & Rashid, 2021):
Where λ is the wavelength of the X-ray = 1.5406 Å, β is the full-width half maximum (FWHM) of the peak in radians, and θ is the Bragg angle.
Figure 3
XRD pattern ofAgO/Fe2O3 NCs preparation from shrimp marine extract by PLA (Nd-YAG) technique.
Table 3
The diffraction angles, Miller indices, full width at half maximum (FWHM), and crystallite size for every diffraction angle.
FE-SEM of AgO/Fe2O3 NCs prepared from marine shrimp shell extract by PLA (Nd-YAG technique
The FE-SEM pictures at an accelerated voltage of 10 kV and a magnification of 50K are shown in Figure 4. AgO/Fe2O3-NCs have an irregular form, as shown in Figure 4(A), and their mean particle size is between 29 and 40 nm. These images display the apparent NP aggregation. Thermodynamics dictates that due to the high power of laser ablation, species collisions in relatively high-energy states should produce NPs soon at the beginning of ablation. In contrast, neutral species aggregations may be the source of nucleation. Figure 4 (B) illustrates how the AgO/Fe2O3-NCs assemble into multi-layered structures after longer ablation durations (Amrillah, 2022; Karpagavinayagam & Vedhi, 2019).
TEM of AgO/Fe2O3 NCs prepared from marine shrimp shell extract by the PLA (Nd-YAG) technique
To identify the morphology and the particle size of AgO/Fe2O3 NCs prepared from marine shrimp extract by PLA (Nd-YAG) technique via TEM device. TEM images show that AgO/Fe2O3 NCs are spherical and aggregated significantly, as shown in Figure 5A–B. The color of AgO/Fe2O3 NCs changed from white to lead dark when using marine shrimp shell extract.
EDS and mapping of AgO/Fe2O3 NCs prepared from marine shrimp shell extract by PLA (Nd-YAG) technique
AgO-Fe2O3 NCs' elemental composition was examined using EDS. Figure 6 and Table 4 illustrate that iron (Fe), oxygen (O), and silver (Ag) were the three prominent peaks. The many phytochemicals included in the extract were the primary cause of the signals for C, N, and K. The presence of an oxygen signal further confirms the creation of iron oxide nanoparticles. For Fe, O, and Ag, the weight percent (wt%) from nanoparticles was determined to be 16.33%, 28.33%, and 24.35%, respectively (Ali, Hussein, & Dakhil, 2021; Elbasuney, El-Sayyad, Attia, & Abdelaziz, 2022). The as-prepared nanoparticles may be readily magnetically recovered due to the high Fe loading. It would result from the marine shrimp extract of the substance. The EDX elemental mapping of a single particle aggregate is shown in Figure 6B. The X-ray signals from Ag, Fe, and O are represented by blue, green, and yellow, respectively. The collected data demonstrates that all of these components' spatial distribution inside the nanocomposite is relatively uniform (Kadhim, Abid, & Salih, 2023; Salih, Sabah, Kadhim, Kadhum, & Abid, 2023).
Figure 6
Mapping and EDS images of AgO/Fe2O3 NCs preparation from the marine shrimp shell extract by PLA (Nd-YAG) technique.
Table 4
The elements, line type, weight, weight sigma, and atomic percentageof AgO/Fe2O3 NCs preparation from the marine shrimp shell extract by PLA (Nd-YAG) technique.
FTIR spectrum of AgO/Fe2O3 NCs prepared from marine shrimp shell extract by PLA (Nd-YAG) technique
The FTIR spectra of AgO/Fe2O3 NCs in the 4000-500 cm−1 range are shown in Figure 7 . The peaks at 3899.61, 3847.18, 3742.40, 3675.89, 3646.54, 3491.07, 3450.86, and 3413.63 cm−1 correspond to the O-H stretching of the free OH group. An alkynes' C-H bond stretching vibrations are responsible for the 3063.64 cm−1 peak. The peak at 2923.77 cm−1 is caused by the amine salt's N-H bond stretching. A signal at 2854.52 cm-1 represents asymmetric stretching vibrations of CH2 in aliphatic hydrocarbons. A 2586.08 and 2367.49 cm−1 peak are attributed to alkynes' C=C stretching vibrations. After phase transfer, a new band at 1843.88 cm−1 develops. The carboxylic acid's C=O bond stretching vibrations are responsible for the 1742.41 cm−1 peak. This corresponds to the symmetric vibration modes of the COO group, indicating chemical interactions between the carboxyl groups. The stretching vibration of an acyclic C-C produced from aromatic rings, shown by the strong peak at 1636.35 cm-1 with 1523.83 and 1456.22, is present in marine shrimp marine shell extract. To the amine group, 1393 cm−1 and 1042 cm−1 assicated. Ag-O bending vibrations (fingerprint) had 785.44 and 821.47 cm−1 peaks, while the Fe-O vibrations (fingerprint) had 785.13 and 726.46 cm−1 peaks, as shown in Figure 7 (Batoo et al., 2024; Jasim, Dakhil, Hussein, & Abdullah, 2020; Kadhim et al., 2022; Satheesh, Vignesh, Suganthi, & Rajarajan, 2014).
Antimicrobial activity of AgO/Fe2O3 NCs prepared from marine shrimp shell extract by PLA (Nd-YAG) technique
The AgO/Fe2O3 NCs made from marine shrimp shell extract using the PLA (Nd-YAG) approach showed antibacterial efficacy against both gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria. The antibacterial activity of marine shrimp shell extract stabilizer-based AgO/Fe2O3-NCs was examined using the agar well diffusion method compared to bacteria (Aldahasi, Shami, & Mohammed, 2024; Chircov et al., 2022; Dewangan, Bhatia, Singh, & Carabineiro, 2024; Ezealigo, Ezealigo, Aisida, & Ezema, 2021; Ghazzy et al., 2024; Li et al., 2019). The inhibition zones of pathogenic gram-negative bacteria (Escherichia coli) and gram-positive bacteria (Staphylococcus aureus and Staphylococcus epidermidis) are displayed in Figure 8(A–C). One well-indicated via AgO/Fe2O3-NCs is loaded with 30 mg/L (AgO/Fe2O3-NCs) (Attia et al., 2022; Edwar, Naji, & Maleki, 2023; Ezealigo, Ezealigo, Aisida, & Ezema, 2021; Zhang et al., 2022). It determined the proportion of destroyed bacteria when AgO/Fe2O3-NCs were used with marine shrimp shell extract. This is because it was evident that the biomolecules in the chard extract had a more significant impact than the biomolecules in the shrimp extract, and the outcome was excellent and distinct from previous findings (Shaymaa, Ismail, Kadhim, Ban, & Yousif, 2023).
The inhibition of zone (%) of AgO/Fe2O3-NCs using marine shrimp extract by a PLA (Nd-YAG) technique at 30 mg/L. With the applied PLA technique, when employing AgO/Fe2O3-NCs from marine shrimp shell extract, it was shown that the percentage of destruction for gram-positive and gram-negative bacteria was good. The results of the destruction of the three types of bacteria and a kind of fungi, as shown in Figure 9; Figure 8 and Table 5 (Ansari et al., 2023; Ejileugha, Ezealisiji, Ezejiofor, & Orisakwe, 2021; Homsi, Al-Said, & Tahan, 2023; Nzilu et al., 2023; Shah & Bharadvaja, 2024; Shaymaa, Ismail, Ali, Hussien, & Alheety, 2023), Because of their ultra-small, controlled size, AgO/Fe2O3 NCs are ideal for performing antibacterial operations and battling internal microorganisms. The nanocomposites' crystallite size, particle size, shape, elemental composition, and functional groups are so tiny that bacteria cells may readily phagocytose them. Furthermore, numerous NC kinds have structures that bacteria can use. (for example, liposomal NCs, whose walls are composed of one or more lipid bilayers surrounding sphere-shaped NCs), and the flexibility of NCs to enter host cells via endocytosis allows the majority of the nano to be released intracellularly (Wang, Hu, & Shao, 2017).
The antifungal activity of Candida was used in the AgO/Fe2O3-NCs preparation of marine shrimp extract using the PLA (Nd-YAG) technique. The antifungal activity of AgO/Fe2O3-NCs from marine shrimp shell extract stabilizer-based was investigated versus Candida albicans by the agar well diffusion technique. It is observed that the effect of AgO/Fe2O3-NCs on Candida is high, as shown in Figure 8D (Abbas, Gdoura, & Al-Marjani, 2024; Carrière & Larue, 2012; Ghelich et al., 2022; Kadhim, Abid, Latif, Salih, & Al-Kazazz, 2024; Prathap et al., 2023).
One of the leading causes of illness in both industrialized and poor countries is thought to be bacterial infection. The pathogens change throughout time, developing resistance to previously found antibiotics. Every year, two million individuals worldwide are afflicted with various types of bacteria, and 700,000 of them die as a result of bacterial resistance. For instance, in the US and Europe, Staphylococcus aureus, Staphylococcus epidermidis, and Escherichia coli are responsible for 50,000 deaths annually. Because of this, specialists are growing more worried about how well-suited contemporary antibiotics are to treating a range of infectious diseases. A lot of research was done on creating new, alternative drugs to treat resistance problems. Nature is a possible source for drug discovery. The biological components and phytochemicals found in plants have long piqued the interest of scientists due to their various structures, low toxicity, and increased human acceptability. There are several of these antimicrobial phytochemicals (Jubair, Rajagopal, Chinnappan, Abdullah, & Fatima, 2021).
Figure 8
Depicts the antimicrobial activity of AgO/Fe2O3 NCs prepared from marine shrimp shell extract by the PLA (Nd-YAG) technique, A) Escherichia coli bacteria, B) Staphylococcus aureus bacteria, C) Staphylococcus epidermidis, and D) Candida albicans.
Figure 9
D) depicts the antimicrobial activity of control (D. W.), A) Escherichia coli bacteria, B) Staphylococcus aureus bacteria, C) Staphylococcus epidermidis, and D) Candida albicans.
Table 5
Results of the antimicrobial activity of AgO/Fe2O3 NCs prepared from marine shrimp shell extract by PLA (Nd-YAG) technique.
Conclusion
This research developed a natural resource marine shrimp shell extract for synthesising AgO/Fe2O3-NCs by the PLA (Nd-YAG) technique at 1500 pulses and 200 Mj. As revealed by XRD analysis, AgO/Fe2O3-NCs have crystallite sizes ranging from 9.6 to 19.5 nm. AgO/Fe2O3-NCs had a particle size and shape of around 10 nm, and FE-SEM examination revealed that the particles had a nano-spherical structure. The AgO/Fe2O3-NCs properties have excellent behavior toward anti-negative, anti-positive bacterial, and antifungal activity. TEM scans indicated that AgO/Fe2O3 NCs were spherical and aggregation-shaped, with sizes ranging from 15.4 to 23 nm. Using EDS analysis, the elemental values of AgO/Fe2O3-NCs were found to be 16.33%, 28.33%, and 24.35%, respectively. According to the FTIR spectra, the prominent absorption peaks of AgO/Fe2O3-NCs are 726 cm-1. AgO/Fe2O3-NCs EDS mapping results show that the spatial distribution of all these components inside the NCs is relatively uniform. Relative to AgO/Fe2O3-NCs, they exhibited the best response to high killing for antibacterial. The inhibition zones of antibacterial agents utilizing AgO/Fe2O3 NCs were determined using the diffusion technique. The inhibition zones measured 29 to 13.5 mm for gram-positive bacteria (Staphylococcus aureus and Staphylococcus epidermidis) and 40 mm for gram-negative bacteria (Escherichia coli); Candida had an inhibition zone diameter (IZD) of 42 mm. The nanocomposite used against bacteria and fungi was the mechanism of effect of the properties. To enhance the inhibitory zone region, the following antibacterial and antifungal characteristics must be changed and improved: Average grain size: When particles are lowered to the nanoscale, their specific surface area and the number of active sites rise.
Conflicts of interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Author contributions
The authors confirm contribution to the paper as follows: study conception and design: Duha A. Kadhim; data collection: Abdul Haleem A. Raheem; analysis and interpretation of results: Nada Qasim; supervisor: Muslim A. Abid; draft manuscript preparation: Ihab Adnan Thabet. All authors reviewed the results and approved the final version of the manuscript.
