مطالعه مقایسه‌ای فعالیت ضدقارچی نانوذرات اکسید روی سنتزشده به روش سبز، عصاره سیر و داروهای ضدقارچی متداول علیه گونه‌های کریپتوکوکوس جداشده از فضولات کبوتر

مقالات آماده انتشار

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه میکروبیولوژی، دانشکده دامپزشکی، دانشگاه دیالی، عراق.

2 گروه پاتولوژی، دانشکده دامپزشکی، دانشگاه دیالی، عراق

3 گروه میکروبیولوژی، دانشکده دامپزشکی، دانشگاه دیالی، عراق

4 گروه زیست‌شناسی، دانشکده علوم، دانشگاه دیالی، عراق

10.22103/jab.2026.27226.1910

چکیده

هدف: Cryptococcus neoformans و Cryptococcus laurentii قارچ‌های بیماری‌زای فرصت‌طلبی هستند که با بیماری کریپتوکوکوزیس ارتباط دارند و فضولات کبوتر یکی از مهم‌ترین مخازن محیطی این میکروارگانیسم‌ها محسوب می‌شود. افزایش مقاومت قارچی و سمیت ناشی از درمان‌های ضدقارچی متداول، ضرورت جستجوی عوامل ضدمیکروبی جایگزین را برجسته ساخته است. هدف این مطالعه، ارزیابی و مقایسه فعالیت ضدقارچی نانوذرات اکسید روی سنتزشده به روش سبز، عصاره سیر و داروهای ضدقارچی رایج علیه گونه‌های Cryptococcus جداشده از فضولات کبوتر بود.
مواد و روش‌ها: در مجموع، 40 نمونه فضولات کبوتر از مناطق مختلف استان دیالی عراق جمع‌آوری شد. جداسازی قارچ‌ها با استفاده از محیط کشت سابورو دکستروز آگار حاوی کلرامفنیکل انجام شد و شناسایی آنها از طریق بررسی میکروسکوپی و سامانه VITEK 2 تأیید گردید. حساسیت ضدقارچی با استفاده از روش انتشار در چاهک آگار و آزمون رقت‌سازی متوالی در محیط مایع ارزیابی شد. زنده‌مانی سلول‌های مخمری با استفاده از شناساگر آلامار بلو مبتنی بر رزازورین بررسی گردید. نانوذرات اکسید روی به روش سنتز سبز و با استفاده از عصاره سیر به‌عنوان عامل احیاکننده و پایدارکننده تولید شدند.
نتایج: از میان نمونه‌های بررسی‌شده، 13 نمونه (32/5 درصد) از نظر گونه‌های Cryptococcus مثبت بودند که از این میان، 61/5 درصد به عنوان C. neoformans و 38/4 درصد به عنوان C. laurentii شناسایی شدند. نانوذرات اکسید روی و عصاره سیر هر دو فعالیت ضدقارچی معنی‌دار و وابسته به غلظت علیه جدایه‌های مورد مطالعه نشان دادند. نانوذرات اکسید روی اثرات مهاری قابل توجهی ایجاد کردند و در غلظت‌های مختلف، هاله‌های عدم رشد و مقادیر متفاوت حداقل غلظت مهارکنندگی مشاهده شد. به‌طور مشابه، عصاره سیر نیز فعالیت ضدقارچی مطلوبی علیه هر دو گونه نشان داد، اگرچه قدرت اثر آن اندکی کمتر از نانوذرات بود. داروهای ضدقارچی متداول از جمله ایتراکونازول و فلوکونازول نیز اثرات مهاری قابل توجهی داشتند، اما الگوهای حساسیت در میان جدایه‌های مختلف متفاوت بود.
نتیجه‌گیری: به طور کلی، نتایج این مطالعه نشان داد که نانوذرات اکسید روی سنتزشده به روش سبز دارای پتانسیل ضدقارچی قوی هستند که احتمالاً ناشی از اندازه نانومتری، سطح ویژه بالا و توانایی آنها در تخریب ساختارهای سلولی قارچ‌ها است. عصاره سیر نیز به واسطه ترکیبات فیتوشیمیایی خود، به‌ویژه آلیسین، فعالیت ضدقارچی قابل توجهی از خود نشان داد. این یافته‌ها بیانگر آن است که فناوری نانوی سبز و ترکیبات مشتق‌شده از گیاهان می‌توانند به عنوان راهبردهای مکمل یا جایگزین امیدوارکننده برای کنترل و درمان عفونت‌های ناشی از Cryptococcus، به‌ویژه در شرایط افزایش مقاومت ضدقارچی، مورد استفاده قرار گیرند.

کلیدواژه‌ها

عنوان مقاله [English]

A comparative study of the antifungal activity of green-synthesized ZnO nanoparticles, garlic extract, and conventional antifungal agents against Cryptococcus spp. isolated from pigeon droppings

نویسندگان [English]

  • Noor Their Talib 1
  • Hiba Yaseen Abbass 2
  • Shahad Abdul Majeed Aswad 3
  • Noor Ali Faleh 4
  • Anaaam Faud Hussain 4
1 Department of Microbiology, College of Veterinary Medicine, University of Diyala, Iraq
2 Department of Pathology, College of Veterinary Medicine, University of Diyala, Iraq
3 Department of Microbiology, College of Veterinary Medicine, University of Diyala, Iraq
4 Department of Biology, College of Science, University of Diyala, Iraq
چکیده [English]

Objective
Cryptococcus neoformans and Cryptococcus laurentii are opportunistic fungal pathogens associated with cryptococcosis, and pigeon droppings serve as a major environmental reservoir for these organisms. Increasing antifungal resistance and toxicity of conventional therapies have prompted the search for alternative antimicrobial agents. This study aimed to evaluate and compare the antifungal activity of green-synthesized zinc oxide nanoparticles (ZnO NPs), garlic (Allium sativum) extract, and conventional antifungal agents against Cryptococcus spp. isolated from pigeon droppings.
Materials and methods
A total of 40 pigeon dropping samples were collected from different locations in Diyala Governorate, Iraq. Fungal isolation was performed using Sabouraud dextrose agar supplemented with chloramphenicol, and identification was confirmed through microscopic examination and the VITEK 2 system. Antifungal susceptibility testing was conducted using the agar well diffusion method and broth microdilution assay. Yeast cell viability was assessed using the resazurin-based Alamar Blue indicator. ZnO nanoparticles were synthesized via a green method using garlic extract as a reducing and stabilizing agent.
Results
Among the examined samples, 13 (32.5%) were positive for Cryptococcus spp., of which 61.5% were identified as C. neoformans and 38.4% as C. laurentii. Both ZnO nanoparticles and garlic extract exhibited significant, concentration-dependent antifungal activity against the tested isolates. ZnO nanoparticles demonstrated notable inhibitory effects, with variable inhibition zones and minimum inhibitory concentration (MIC) values ranging across tested concentrations. Similarly, garlic extract showed appreciable antifungal activity against both species, although with slightly lower potency compared to nanoparticles. Conventional antifungal agents, including itraconazole and fluconazole, also showed inhibitory effects; however, variability in susceptibility patterns was observed among isolates.
Conclusion
Overall, the results indicate that green-synthesized ZnO nanoparticles possess strong antifungal potential, likely due to their nanoscale size, high surface area, and ability to disrupt fungal cellular structures. Garlic extract also contributes antifungal bioactivity through its phytochemical constituents, particularly allicin. The combined findings suggest that both green nanotechnology and plant-derived compounds may serve as promising complementary or alternative strategies for managing Cryptococcus infections, particularly in the context of emerging antifungal resistance.

کلیدواژه‌ها [English]

  • antifungal activity
  • Cryptococcus laurentii
  • Cryptococcus neoformans
  • garlic extract
  • zinc oxide nanoparticles

مراجع

Al-Huthaifi, A. M., Radman, B. A., Al-Alawi, A. A., Mahmood, F., & Liu, T.-B. (2024). Mechanisms and virulence factors of Cryptococcus neoformans dissemination to the central nervous system. Journal of Fungi, 10(8), Article 586. https://doi.org/10.3390/jof10080586
Almeida, F., Wolf, J. M., & Casadevall, A. (2015). Virulence-associated enzymes of Cryptococcus neoformans. Eukaryotic Cell, 14(12), 1173–1185. https://doi.org/10.1128/EC.00103-15
Al-Shammary, M. A. A., & Al-Mayaly, I. K. (2024). Green synthesis of zinc oxide nanoparticles from Lyngbya sp. and their antibacterial activity. Iraqi Journal of Science, 65(8), 4281–4290. https://doi.org/10.24996/ijs.2024.65.8.13
Arora, D. R., & Arora, B. B. (2024). Medical mycology (3rd ed.). CBS Publishers & Distributors Pvt. Ltd.
Arreola, R., Quintero-Fabián, S., López-Roa, R. I., Flores-Gutiérrez, E. O., Reyes-Grajeda, J. P., Carrera-Quintanar, L., & Ortuño-Sahagún, D. (2015). Immunomodulation and anti-inflammatory effects of garlic compounds. Journal of Immunology Research, 2015, Article 401630. https://doi.org/10.1155/2015/401630
Arsic Arsenijevic, V., Pekmezovic, M. G., Meis, J. F., & Hagen, F. (2014). Molecular epidemiology and antifungal susceptibility of Serbian Cryptococcus neoformans isolates. Mycoses, 57(6), 380–387. https://doi.org/10.1111/myc.12171
Baigorria, C. G., Cerioni, L., Debes, M. A., Ledesma, A. E., Alastuey, P., Tirado, M., Volentini, S. I., & Rapisarda, V. A. (2024). Antifungal action of metallic nanoparticles against fungicide-resistant pathogens causing main postharvest lemon diseases. Journal of Fungi, 10(11), Article 782. https://doi.org/10.3390/jof10110782
Balouiri, M., Sadiki, M., & Ibnsouda, S. K. (2016). Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis, 6(2), 71–79. https://doi.org/10.1016/j.jpha.2015.11.005
Borlinghaus, J., Albrecht, F., Gruhlke, M. C. H., Nwachukwu, I. D., & Slusarenko, A. J. (2014). Allicin: Chemistry and biological properties. Molecules, 19(8), 12591–12618. https://doi.org/10.3390/molecules190812591
Bouttier-Figueroa, D. C., Cortez-Valadez, M., Flores-Acosta, M., & Robles-Zepeda, R. E. (2024). Green synthesis of zinc oxide nanoparticles using plant extracts and their antimicrobial activity. BioNanoScience, 14, 3385–3400. https://doi.org/10.1007/s12668-024-01385-5
Briner, S. L., & Doering, T. L. (2025). Cryptococcus. Current Biology, 35(11), R518–R522. https://doi.org/10.1016/j.cub.2025.04.008
Brito, M. O., Bessa, M. A. S., Menezes, R. P., Röder, D. V. D. B., Penatti, M. P. A., Pimenta, J. P., Aguiar, P. A. D. F., & Pedroso, R. D. S. (2019). Isolation of Cryptococcus species from the external environments of hospital and academic areas. Journal of Infection in Developing Countries, 13(6), 545–553. https://doi.org/10.3855/jidc.10849
Bunaciu, A. A., Udriştioiu, E. G., & Aboul-Enein, H. Y. (2015). X-ray diffraction: Instrumentation and applications. Critical Reviews in Analytical Chemistry, 45(4), 289–299. https://doi.org/10.1080/10408347.2014.949616
Canónico-González, Y., Adame-Rodríguez, J. M., Mercado-Hernández, R., & Aréchiga-Carvajal, E. T. (2013). Cryptococcus spp. isolation from excreta of pigeons (Columba livia) in and around Monterrey, Mexico. SpringerPlus, 2(1), Article 632. https://doi.org/10.1186/2193-1801-2-632
Chong, H. S., Dagg, R., Malik, R., Chen, S., & Carter, D. (2010). In vitro susceptibility of the yeast pathogen Cryptococcus to fluconazole and other azoles varies with molecular genotype. Journal of Clinical Microbiology, 48(11), 4115–4120. https://doi.org/10.1128/JCM.01271-10
Clinical and Laboratory Standards Institute. (2026). Performance standards for antimicrobial susceptibility testing (M100, 36th ed.). https://clsi.org/shop/standards/m100/
Daoud, A., Malika, D., Bakari, S., Hfaiedh, N., Mnafguia, K., Kadri, A., & Gharsallah, N. (2015). Assessment of polyphenol composition, antioxidant and antimicrobial properties of various extracts of Date Palm Pollen (DPP) from two Tunisian cultivars. Arabian Journal of Chemistry, 12(8), 3075–3086. https://doi.org/10.1016/j.arabjc.2015.07.014
Davis, L. E., Shen, J., & Royer, R. E. (1994). In vitro synergism of concentrated Allium sativum extract and amphotericin B against Cryptococcus neoformans. Planta Medica, 60(6), 546–549. https://doi.org/10.1055/s-2006-959568
Dorjee, L., Gogoi, R., Kamil, D., Kumar, R., Mondal, T. K., Pattanayak, S., & Gurung, B. (2023). Essential oil-grafted copper nanoparticles as a potential next-generation fungicide for holistic disease management in maize. Frontiers in Microbiology, 14, Article 1204512. https://doi.org/10.3389/fmicb.2023.1204512
Eloff, J. N. (1998). Which extractant should be used for the screening and isolation of antimicrobial components from plants? Journal of Ethnopharmacology, 60(1), 1–8. https://doi.org/10.1016/S0378-8741(97)00123-2
El-Saber Batiha, G., Magdy Beshbishy, A., G. Wasef, L., Elewa, Y. H. A., A. Al-Sagan, A., Abd El-Hack, M. E., Taha, A. E., M. Abd-Elhakim, Y., & Prasad Devkota, H. (2020). Chemical constituents and pharmacological activities of garlic (Allium sativum L.): A review. Nutrients, 12(3), Article 872. https://doi.org/10.3390/nu12030872
Gonelimali, F. D., Lin, J., Miao, W., Xuan, J., Charles, F., Chen, M., & Hatab, S. R. (2018). Antimicrobial properties and mechanism of action of some plant extracts against food pathogens and spoilage microorganisms. Frontiers in Microbiology, 9, Article 1639. https://doi.org/10.3389/fmicb.2018.01639
Herkert, P. F., Meis, J. F., Lucca de Oliveira Salvador, G., Rodrigues Gomes, R., Aparecida Vicente, V., Dominguez Muro, M., Lameira Pinheiro, R., Lopes Colombo, A., Vargas Schwarzbold, A., Sakuma de Oliveira, C., Simão Ferreira, M., Queiroz-Telles, F., & Hagen, F. (2018). Molecular characterization and antifungal susceptibility testing of Cryptococcus neoformans sensu stricto from southern Brazil. Journal of Medical Microbiology, 67(4), 560–569. https://doi.org/10.1099/jmm.0.000698
Ikeda, R., Sugita, T., Jacobson, E. S., & Shinoda, T. (2003). Effects of melanin upon susceptibility of Cryptococcus to antifungals. Microbiology and Immunology, 47(4), 271–277. https://doi.org/10.1111/j.1348-0421.2003.tb03395.x
Jain, N., Jain, P., Rajput, D., Patil, U. K. (2021). Green synthesized plant-based silver nanoparticles: Therapeutic prospective for anticancer and antiviral activity. Micro and Nano Systems Letters, 9, Article 5. https://doi.org/10.1186/s40486-021-00131-6
Kamal, I. M., & Al-haddad, Z. A. A. (2022). Isolation and molecular identification of pathogenic Cryptococcus neoformans from pigeon dropping and human sputum. International Journal of Health Sciences, 6(S4), 8363–8374. https://doi.org/10.53730/ijhs.v6nS4.10554
Khadka, D., Bista, P., Baral, J., Gautam, S. K., Bastakoti, B. P., Poudel, B. R., & Pokhrel, M. R. (2025). Green synthesis of zinc oxide nanoparticles using Swertia chirayita for photocatalytic and antimicrobial activity. Journal of Institute of Science and Technology, 30(1), 45–56. https://doi.org/10.3126/jist.v30i1.72209
Kidd, S., Halliday, C., & Ellis, D. (2026). Description of medical fungi. CABI.
Kwon-Chung, K. J., Fraser, J. A., Doering, T. L., Wang, Z., Janbon, G., Idnurm, A., & Bahn, Y. S. (2014). Cryptococcus neoformans and Cryptococcus gattii, the etiologic agents of cryptococcosis. Cold Spring Harbor Perspectives in Medicine, 4(7), Article a019760. https://doi.org/10.1101/cshperspect.a019760
Mahmoudi, S., Mamishi, S., Banar, M., Pourakbari, B., & Keshavarz, H. (2019). Epidemiology of echinococcosis in Iran: A systematic review and meta-analysis. BMC Infectious Diseases, 19(1), Article 929. https://doi.org/10.1186/s12879-019-4458-5
Mohammadabadi, M. R., & Mozafari, M. R. (2019). Development of nanoliposome-encapsulated thymoquinone: Evaluation of loading efficiency and particle characterization. Journal of Biopharmaceuticals, 11(4), 39–46.
Mohammadabadi, M. R., El-Tamimy, M., Gianello, R., & Mozafari, M. R. (2009). Supramolecular assemblies of zwitterionic nanoliposome-polynucleotide complexes as gene transfer vectors: Nanolipoplex formulation and in vitro characterization. Journal of Liposome Research, 19(2), 105–115. https://doi.org/10.1080/08982100802547326
Montagna, M. T., De Donno, A., Caggiano, G., Serio, F., De Giglio, O., Bagordo, F., D’Amicis, R., Lockhart, S. R., & Cogliati, M. (2018). Molecular characterization of Cryptococcus neoformans and Cryptococcus gattii from environmental sources and genetic comparison with clinical isolates in Apulia, Italy. Environmental Research, 160, 347–352. https://doi.org/10.1016/j.envres.2017.09.032
Montenegro, H., & Paula, C. R. (2000). Environmental isolation of Cryptococcus neoformans var. gattii and C. neoformans var. neoformans in the city of São Paulo, Brazil. Medical Mycology, 38(5), 385–390. https://doi.org/10.1080/mmy.38.5.385.390
Na Pombejra, S., Salemi, M., Phinney, B. S., & Gelli, A. (2017). The metalloprotease, Mpr1, engages AnnexinA2 to promote the transcytosis of fungal cells across the blood-brain barrier. Frontiers in Cellular and Infection Microbiology, 7, Article 296. https://doi.org/10.3389/fcimb.2017.00296
Olszewski, M. A., Noverr, M. C., Chen, G. H., Toews, G. B., Cox, G. M., Perfect, J. R., & Huffnagle, G. B. (2004). Urease expression by Cryptococcus neoformans promotes microvascular sequestration, thereby enhancing central nervous system invasion. The American Journal of Pathology, 164(5), 1761–1771. https://doi.org/10.1016/S0002-9440(10)63734-0
Parekh, J., & Chanda, S. (2007). In vitro antimicrobial activity and phytochemical analysis of some Indian medicinal plants. Turkish Journal of Biology, 31(1), 53–58.
Pincus, I., & Qi, Q. M. (2025). Nanoparticle-induced lipid membrane deformation influences the design of biomedicine. Biophysical Journal. Advance online publication. https://doi.org/10.1016/j.bpj.2025.10.030
Rocha, F. S., Gomes, A. J., Lunardi, C. N., Kaliaguine, S., & Patience, G. S. (2018). Experimental methods in chemical engineering: Ultraviolet visible spectroscopy—UV-Vis. The Canadian Journal of Chemical Engineering, 96(12), 2512–2517. https://doi.org/10.1002/cjce.23344
Shang, A., Cao, S.-Y., Xu, X.-Y., Gan, R.-Y., Tang, G.-Y., Corke, H., Mavumengwana, V., & Li, H.-B. (2019). Bioactive compounds and biological functions of garlic (Allium sativum L.): A review. Foods, 8(7), Article 246. https://doi.org/10.3390/foods8070246
Sionov, E., Chang, Y. C., & Kwon-Chung, K. J. (2013). Azole heteroresistance in Cryptococcus neoformans: Emergence of resistant clones with chromosomal disomy in the mouse brain during fluconazole treatment. Antimicrobial Agents and Chemotherapy, 57(10), 5127–5130. https://doi.org/10.1128/AAC.00694-13
Soltani, M., Bayat, M., Hashemi, S. J., Zia, M., & Pestechian, N. (2013). Isolation of Cryptococcus neoformans from pigeon droppings in Ahwaz, Iran. Turkish Journal of Medical Sciences, 40(2), 313–316. https://doi.org/10.3906/sag-0808-10
Trpković, A., Pekmezović, M., Barać, A., Crnčević Radović, L., & Arsić Arsenijević, V. (2012). In vitro antifungal activities of amphotericin B, 5-fluorocytosine, fluconazole and itraconazole against Cryptococcus neoformans isolated from cerebrospinal fluid and blood from patients in Serbia. Journal de Mycologie Médicale, 22(3), 243–248. https://doi.org/10.1016/j.mycmed.2012.06.002
Walsh, T. J., Hayden, R. T., & Larone, D. H. (2018). Larone's medically important fungi: A guide to identification. ASM Press.
Wu, Q., Cen, F., Xie, Y., Ning, X., Wang, J., Lin, Z., & Huang, J. (2025). Nanoparticle-based antifungal therapies: Innovations, mechanisms and future prospects. PeerJ, 13, Article e19199. https://doi.org/10.7717/peerj.19199
Yamada, Y., & Azuma, K. (1977). Evaluation of the in vitro antifungal activity of allicin. Antimicrobial Agents and Chemotherapy, 11(4), 743–749. https://doi.org/10.1128/AAC.11.4.743
Yang, C. L., Wang, J., & Zou, L. L. (2017). Innate immune evasion strategies against cryptococcal meningitis caused by Cryptococcus neoformans. Experimental and Therapeutic Medicine, 14(6), 5243–5250. https://doi.org/10.3892/etm.2017.5220
Yoshida, S., Kasuga, S., Hayashi, N., Ushiroguchi, T., Matsuura, H., & Nakagawa, S. (1987). Antifungal activity of ajoene derived from garlic. Applied and Environmental Microbiology, 53(3), 615–617. https://doi.org/10.1128/aem.53.3.615-617.1987
Zarrabi, A., Alipoor Amro Abadi, M., Khorasani, S., Mohammadabadi, M., Jamshidi, A., Torkaman, S., Taghavi, E., Mozafari, M. R., & Rasti, B. (2020). Nanoliposomes and tocosomes as multifunctional nanocarriers for the encapsulation of nutraceutical and dietary molecules. Molecules, 25(3), Article 638. https://doi.org/10.3390/molecules25030638

Zarrin, M., Jorfi, M., Amirrajab, N., & Rostami, M. (2010). Isolation of Cryptococcus neoformans from pigeon droppings in Ahwaz, Iran. Turkish Journal of Medical Sciences, 40(2), 313–316. https://doi.org/10.3906/sag-0808-10

مجله بیوتکنولوژی کشاورزی

مقالات آماده انتشار، پذیرفته شده
انتشار آنلاین از تاریخ 12 تیر 1405

فایل ها

هم رسانی

ارجاع به این مقاله

آمار