Agricultural Biotechnology Journal

Agricultural Biotechnology Journal

Experimental and In Silico evaluation of a novel ZnO/Alhagi maurorum/PVA composite film against multidrug-resistant Pseudomonas aeruginosa harboring blaTEM and blaIMP genes

Document Type : Research Paper

Authors
Nisreen Kaddim Radi 1
Shireen Sabah Kamil 2
Safa Hasan Radhi 3
Hussein J. Hussein 2
Ali H. Al-Marzoqi 2
Lubna Abdulazeem 4
Israa A. Ibraheam 2
Ali Salam Kashkool 5
1 Department of Biology, College of Science for Women, University of Babylon, Iraq.
2 Department of Biology, College of science for women, University of Babylon, Iraq.
3 Biology Department, College of science, Al-Qasim Green University, Babylon 51013, Iraq
4 DNA Research center, University of Babylon, Iraq.
5 Middle Technical University, Kut Technical Institute, Iraq
10.22103/jab.2026.27227.1909
Abstract
Objective
A major contributor to burn infections, Pseudomonas aeruginosa is known for its aggressiveness and medication resistance. This study was carried out to prepare and characterize ZnONPs/PVA (nanoparticles) film loaded with Alhagi maurorum extract for multidrug-resistant Pseudomonas aeruginosa isolated from skin burn infections, as well as to molecularly detect and sequence blaTEM and blaIMP genes followed by an in-silico docking analysis to investigate the antibacterial mechanism.

Materials and methods
Alhagi maurorum shoots were collected. Voucher specimens were retained. The GC-MS analysis was performed. Zinc oxide nanoparticles (ZnO-NPs) were used in the polymer matrix. Comprehensive assessments were conducted to demonstrate the successful entrapment and homogeneous distribution of bioactivating nanocomponents in the polymeric matrix of the fabricated composite film. Fifty bacterial isolates were obtained from burn swabs that had been isolated from the burn unit at AL-Sadiq Teaching Hospital, Babylon, Iraq. The antimicrobial susceptibility of P. aeruginosa isolates (n = 20) was determined. Genomic DNA from bacterial isolates was isolated and PCR was performed. The nucleotide sequences of the blaTEM gene from P. aeruginosa were acquired from the National Center for Biotechnology Information (NCBI). SPSS software was used for statistical analysis.
Results
GC-MS revealed that the mass spectrum and structure of twelve chemical components extracted from Alhagi maurorum using methanol are displayed in the GC-MS data. Among them, 9,12-Octadecadienoic acid (Z,Z)-(Linoleic) was unique. By incorporating these phytochemicals into a PVA polymer matrix with zinc nanoparticles, we created a film that decomposes naturally. The film was tested against medical isolates of Pseudomonas aeruginosa, which are drug-resistant bacteria. Among 50 bacterial isolates, 20 were identified as P. aeruginosa. PCR analysis revealed the presence of blaTEM and blaIMP genes in a substantial proportion of the isolates. Molecular docking results suggest that the ZnO nanostructure has the highest binding potential toward the 4b7q protein compared to linoleic acid and PVA. The strong binding affinity and low inhibition constant of ZnO highlight its potential role as a stable binder or inhibitor, whereas linoleic acid and PVA are less effective.
Conclusion: The strong antibacterial properties of the composite film are probably the result of the PVA polymer, zinc nanoparticles, and plant extract working in concert. The ZnO/Alhagi/PVA composite film demonstrated promising antibacterial and antioxidant activities and may represent a potential biodegradable wound-dressing material for managing MDR P. aeruginosa infections.
Keywords
Alhagi marorum
antibacterial films
beta-lactamase
polyvinyl alcohol
zinc nanoparticles
Abazari, M., Badeleh, S. M., Barkhordari, S., Mohammed, S. S., Ahmed, M. M., Mohammed Ameen, M. S., & Abdollahi, H. (2025). Preparation and characterization of polyvinyl alcohol thin films: Rheological, physicochemical and functional properties. Heliyon, 11(12), Article e43604. https://doi.org/10.1016/j.heliyon.2025.e43604
Abdelrazek, E. M., Elashmawi, I. S., & Labeeb, S. (2010). Chitosan filler effects on the experimental characterization, spectroscopic investigation and thermal studies of PVA/PVP blend films. Physica B: Condensed Matter, 405(8), 2021-2027. https://doi.org/10.1016/j.physb.2010.01.095
Abdelrazek, E. M., Abdelghany, A. M., & Tarabiah, A. E. (2019). AC conductivity and dielectric characteristics of PVA/PVP nanocomposite filled with MWCNTs. Journal of Materials Science: Materials in Electronics, 30, 15521-15533. https://doi.org/10.1007/s10854-019-01929-2
Abdulbary, M. (2019). Study of some properties and antibacterial activity of (Alhagi maurorum Medik) collected from the desert of Al-Najaf. Journal of Physics: Conference Series, 1294, Article 062081. https://doi.org/10.1088/1742-6596/1294/6/062081
Ahmed, S., Ahmad, M., Swami, B. L., & Ikram, S. (2016). A review on plants extract mediated synthesis of silver nanoparticles for antimicrobial applications: A green expertise. Journal of Advanced Research, 7(1), 17-28. https://doi.org/10.1016/j.jare.2015.02.007
Al-Duhaidahawi, D., Al-Zubaidy, H. F. S., Al-Khafaji, K., & Al-Amiery, A. (2022). Synthesis, anti-inflammatory effects, molecular docking and molecular dynamics studies of 4-hydroxy coumarin derivatives as inhibitors of COX-II enzyme. Journal of Molecular Structure, 1247, Article 131377. https://doi.org/10.1016/j.molstruc.2021.131377
Al-Snafi, A. (2015). Alhagi maurorum as a potential medicinal herb: An overview. International Journal of Pharmacy Review and Research, 5, 130-136.
Asadpour, S., Raeisi vanani, A., Kooravand, M., & Asfaram, A. (2022). A review on zinc oxide/poly(vinyl alcohol) nanocomposites: Synthesis, characterization and applications. Journal of Cleaner Production, 362, Article 132297. https://doi.org/10.1016/j.jclepro.2022.132297
Barbălată-Mândru, M., Serbezeanu, D., Butnaru, M., Rîmbu, C. M., Enache, A. A., & Aflori, M. (2022). Poly(vinyl alcohol)/plant extracts films: Preparation, surface characterization and antibacterial studies against Gram positive and Gram negative bacteria. Materials, 15(7), Article 2493. https://doi.org/10.3390/ma15072493
Benson, D. A., Cavanaugh, M., Clark, K., Karsch-Mizrachi, I., Lipman, D. J., Ostell, J., & Sayers, E. W. (2013). GenBank. Nucleic Acids Research, 41(D1), D36-D42. https://doi.org/10.1093/nar/gks1195
Brzęk, P., & Sterzyński, T. (2020). Stress whitening as an observation method of residual stress in MABS polymer material through the example of holding pressure in an injection molding process. Polymers, 12(12), Article 2871. https://doi.org/10.3390/polym12122871
Bush, K., & Bradford, P. A. (2020). Epidemiology of β-lactamase-producing pathogens. Clinical Microbiology Reviews, 33(2), Article e00047-19. https://doi.org/10.1128/CMR.00047-19
Castanheira, M., Simner, P. J., & Bradford, P. A. (2021). Extended-spectrum β-lactamases: An update on their characteristics, epidemiology and detection. JAC-Antimicrobial Resistance, 3(3), Article dlab092. https://doi.org/10.1093/jacamr/dlab092
Eftekhari, F., Sarcheshmehpour, M., Lohrasbi-Nejad, A., & Boroomand, N. (2025). Effects of mycorrhizal and Trichoderma treatment on enhancing maize tolerance to salinity and drought stress, through metabolic and enzymatic evaluation. BMC Plant Biology, 25(1), Article 687. https://doi.org/10.1186/s12870-025-06729-x
El-Hachem, N., Haibe-Kains, B., Khalil, A., Kobeissy, F. H., & Nemer, G. (2017). AutoDock and AutoDockTools for protein-ligand docking: Beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) as a case study. Methods in Molecular Biology, 1598, 391-403. https://doi.org/10.1007/978-1-4939-6952-4_20
Franco, D., Calabrese, G., Guglielmino, S. P. P., & Conoci, S. (2022). Metal-based nanoparticles: Antibacterial mechanisms and biomedical application. Microorganisms, 10(9), Article 1778. https://doi.org/10.3390/microorganisms10091778
Ghasemi, H., Omranifard, M., Bahrami, M., Moghimian, M., & Farzi, S. (2025). Challenges in burn care management: A qualitative study of health professionals' and patients' perspectives. Scientific Reports, 15(1), Article 33401. https://doi.org/10.1038/s41598-025-18722-3
Gulcin, İ., & Alwasel, S. H. (2023). DPPH radical scavenging assay. Processes, 11(8), Article 2248. https://doi.org/10.3390/pr11082248
Hakeem, H. S., Muhsen, M. F., & Taieh, N. K. (2024). Optical characterization of polyvinyl alcohol-zinc oxide nanocomposite films fabricated via spray coating and enhanced through plasma modification: A comprehensive study. Journal of Nanotechnology, 2024, Article 8878626. https://doi.org/10.1155/jnt/8878626
Hussein, A. A., Kaddim Radi, N., & Mohammed Sahi, N. (2025). Integrative computational and experimental study of propolis, polyvinyl alcohol, and Alhagi maurorum complex as anticancer and antibacterial agents. Journal of Biomaterials Science, Polymer Edition, 36(12), 1718-1748. https://doi.org/10.1080/09205063.2025.2464448
Ibrahim, S., Al-Dahir, S., Al Mulla, T., Lami, F., Hossain, S. M. M., Baqui, A., & Burnham, G. (2021). Resilience of health systems in conflict affected governorates of Iraq, 2014-2018. Conflict and Health, 15(1), Article 76. https://doi.org/10.1186/s13031-021-00412-2
Jabalia, N., Kumar, A., Kumar, V., & Rani, R. (2021). In silico approach in drug design and drug discovery: An update. In S. K. Singh (Ed.), Innovations and implementations of computer aided drug discovery strategies in rational drug design (pp. 1-28). Springer. https://doi.org/10.1007/978-981-15-8936-2_10
Liang, X., Zhong, H.-J., Ding, H., Yu, B., Ma, X., Liu, X., Chong, C.-M., & He, J. (2024). Polyvinyl alcohol (PVA)-based hydrogels: Recent progress in fabrication, properties, and multifunctional applications. Polymers, 16(19), Article 2755. https://doi.org/10.3390/polym16192755
Mostafa, R. M., & Essawy, H. S. (2019). Assessment of camel thorn (Alhagi maurorum) as new sources of bioactive compounds using GC-MS technique. Plant Omics, 12(1), 70-76.
Muhammad, M., Waheed, A., Wahab, A., Majeed, M., Nazim, M., Liu, Y.-H., Li, L., & Li, W.-J. (2024). Soil salinity and drought tolerance: An evaluation of plant growth, productivity, microbial diversity, and amelioration strategies. Plant Stress, 11, Article 100319. https://doi.org/10.1016/j.stress.2023.100319
Nandy, S., Das, A. K., & Dudeja, M. (2017). Prevalence of metallo beta lactamase in clinical isolates of Pseudomonas aeruginosa in a tertiary care hospital. International Journal of Community Medicine and Public Health, 2(4), 566-569. https://doi.org/10.18203/2394-6040.ijcmph20151048
O'Boyle, N. M., Banck, M., James, C. A., Morley, C., Vandermeersch, T., & Hutchison, G. R. (2011). Open Babel: An open chemical toolbox. Journal of Cheminformatics, 3, Article 33. https://doi.org/10.1186/1758-2946-3-33
Oliver, A., Arca-Suárez, J., Gomis-Font, M. A., González-Pinto, L., López-Causapé, C., & Others. (2025). Emerging resistance mechanisms to newer β-lactams in Pseudomonas aeruginosa. Clinical Microbiology and Infection, 31(11), 1790-1796. https://doi.org/10.1016/j.cmi.2025.03.013
Premkumar, H. (2024). Synthesis, characterization and photocatalytic performance of polyvinyl alcohol/zinc oxide nanocomposites: A comprehensive study. Nano-Structures & Nano-Objects, 39, Article 101208. https://doi.org/10.1016/j.nanoso.2024.101208
Qin, S., Xiao, W., Zhou, C., Pu, Q., Deng, X., Lan, L., Liang, H., Song, X., Wu, M., & Others. (2022). Pseudomonas aeruginosa: Pathogenesis, virulence factors, antibiotic resistance, interaction with host, technology advances and emerging therapeutics. Signal Transduction and Targeted Therapy, 7(1), Article 199. https://doi.org/10.1038/s41392-022-01056-1
Radi, N. K., & Al-Marzoqi, A. H. (2022). Detection of virulent genetic markers like blaTEM, blaIMP1 of pathogenic E. coli strain isolated from Iraqi frozen chicken meat. International Journal of Health Sciences, 6(S1), 6864-6874. https://doi.org/10.53730/ijhs.v6nS1.6679
Rafi, M. O., Al-Khafaji, K., Mandal, S. M., Meghla, N. S., Biswas, P. K., Rahman, M. S., & Others. (2023). A subunit vaccine against pneumonia: Targeting Streptococcus pneumoniae and Klebsiella pneumoniae. Network Modeling Analysis in Health Informatics and Bioinformatics, 12(1), Article 21. https://doi.org/10.1007/s13721-023-00416-3
Saber, T. M., Abo-Elmaaty, A. M. A., Said, E. N., Beheiry, R. R., Moselhy, A. A., Abdelgawad, F. E., Arisha, M. H., Saber, T., Arisha, A. H., & Fahmy, E. M. (2022). Alhagi maurorum ethanolic extract rescues hepato-neurotoxicity and neurobehavioral alterations induced by lead in rats via abrogating oxidative stress and the caspase-3-dependent apoptotic pathway. Antioxidants, 11(10), Article 1992. https://doi.org/10.3390/antiox11101992
Sabr, O. H., Obaid, M. N., Al-Mutairi, N. H., Krynke, M., & Mielczarek, K. (2024). Study the effect of zinc oxide nanoparticles on degradation, antibacterial, thermal and morphological properties of polyvinyl alcohol films. Production Engineering Archives, 30(4), 528-536. https://doi.org/10.30657/pea.2024.30.49
Shalmashi, H., Farajnia, S., Sadeghi, M., Tanoumand, A., Veissi, K., Hamishekar, H., & Gotaslou, R. (2022). Detection of ESBLs types blaCTX-M, blaSHV and blaTEM resistance genes among clinical isolates of Pseudomonas aeruginosa. Gene Reports, 28, Article 101637. https://doi.org/10.1016/j.genrep.2022.101637
Singh, S., Sahoo, R. K., & Sahu, M. C. (2025). Understanding recent developments in colistin resistance: Mechanisms, clinical implications, and future perspectives. Antibiotics, 14(10), Article 958. https://doi.org/10.3390/antibiotics14100958
Sirelkhatim, A., Mahmud, S., Seeni, A., Kaus, N. H. M., Ann, L. C., Bakhori, S. K. M., Hasan, H., & Mohamad, D. (2015). Review on zinc oxide nanoparticles: Antibacterial activity and toxicity mechanism. Nano-Micro Letters, 7, 219-242. https://doi.org/10.1007/s40820-015-0040-x
Tacconelli, E., Carrara, E., Savoldi, A., Harbarth, S., Mendelson, M., Monnet, D. L., Pulcini, C., Kahlmeter, G., Kluytmans, J., Carmeli, Y., Ouellette, M., Outterson, K., Patel, J., Cavaleri, M., Cox, E. M., Houchens, C. R., Grayson, M. L., Hansen, P., Singh, N., ... Magrini, N. (2018). Discovery, research, and development of new antibiotics: The WHO priority list of antibiotic-resistant bacteria and tuberculosis. The Lancet Infectious Diseases, 18(3), 318-327. https://doi.org/10.1016/S1473-3099(17)30753-3
Tamura, K., Stecher, G., Peterson, D., Filipski, A., & Kumar, S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution, 30(12), 2725-2729. https://doi.org/10.1093/molbev/mst197
Türker, F., Noma, S. A. A., Aktaş, A., Al-Khafaji, K., Taşkın Tok, T., Ateş, B., & Gök, Y. (2020). The (NHC)PdBr₂(2-aminopyridine) complexes: Synthesis, characterization, molecular docking study, and inhibitor effects on the human serum carbonic anhydrase and serum bovine xanthine oxidase. Monatshefte für Chemie - Chemical Monthly, 151, 1557-1567. https://doi.org/10.1007/s00706-020-02687-2
Udayagiri, H., Sana, S. S., Dogiparthi, L. K., Vadde, R., Varma, R. S., Koduru, J. R., Ghodake, G. S., Somala, A. R., Boya, V. K. N., Kim, S. C., & Karri, R. R. (2024). Phytochemical fabrication of ZnO nanoparticles and their antibacterial and anti-biofilm activity. Scientific Reports, 14(1), Article 19714. https://doi.org/10.1038/s41598-024-69044-9
Vasile, C., & Baican, M. (2021). Progresses in food packaging, food quality, and safety—Controlled-release antioxidant and/or antimicrobial packaging. Molecules, 26(5), Article 1263. https://doi.org/10.3390/molecules26051263
Wen, Y. H., Tsou, C. H., de Guzman, M. R., Huang, D., Yu, Y. Q., Gao, C., Zhang, X. M., Du, J., Zheng, Y. T., Zhu, H., & Wang, Z. H. (2022). Antibacterial nanocomposite films of poly(vinyl alcohol) modified with zinc oxide-doped multiwalled carbon nanotubes as food packaging. Polymer Bulletin, 79, 3847-3866. https://doi.org/10.1007/s00289-021-03666-1
Wong, T. T., Amigues, S., & Awaja, F. (2025). The influence of zinc oxide nanoparticles on dispersion, rheology, and mechanical properties of epoxy-based composites. Polymers, 17(24), Article 3253. https://doi.org/10.3390/polym17243253
Xie, Y., Cai, P., Cao, X., Chen, B., & Pan, Y. (2024). Water-resistant poly(vinyl alcohol)/ZnO nanopillar composite films for antibacterial packaging. ACS Omega, 9(51), 50403-50413. https://doi.org/10.1021/acsomega.4c07173
Yue, Y., Cheng, X., Liu, H., Zang, M., Zhao, B., Zhao, X., & Wang, L. (2023). Gellan gum and polyvinyl alcohol based triple-layer films enriched with Alhagi sparsifolia flower extract: Preparation, characterization, and application of dried shrimp preservation. Foods, 12(21), Article 3979. https://doi.org/10.3390/foods12213979
Agricultural Biotechnology Journal
Volume 18, Issue 4
Summer 2026
Pages 339-372