آسیب‌شناسی بافت روده در فیل‌ماهی Huso huso پرورشی تغذیه شده با نانوذرات سلنیوم و آهن

نوع مقاله : مقاله پژوهشی

نویسندگان

انستیتو تحقیقات بین المللی ماهیان خاویاری، موسسه تحقیقات علوم شیلاتی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی (AREEO)، رشت، گیلان، ایران

چکیده

این مطالعه با هدف بررسی تأثیر سطوح مختلف نانوذرات سلنیوم (Se-NPs) و نانوذرات آهن (Fe-NPs) بر ساختار بافت روده فیل­ماهی (Huso huso) پرورشی انجام شد. تعداد 135 عدد فیل­ماهی پرورشی (با میانگین وزنی 3/32 ± 4/276 گرم و میانگین درازای کل 2 ± 40 سانتی متر) به طور تصادفی در 27 مخزن فایبر­گلاس در 9 تیمار مختلف با 3 تکرار توزیع شدند. تیمارها شامل تیمار 1: شاهد (بدون افزودن نانوذرات)، تیمار 2: mg/kg Se-NPs 1، تیمار 3: mg/kg Se-NPs 2، تیمار 4: mg/kg Fe-NPs 100، تیمار 5: mg/kg Fe-NPs 100 + mg/kg Se-NPs 1، تیمار 6: mg/kg Fe-NPs 100 + mg/kg Se-NPs 2، تیمار 7: mg/kg Fe-NPs 200، تیمار 8: mg/kg Fe-NPs 200 + mg/kg Se-NPs 1، تیمار 9: mg/kg Fe-NPs 200 + mg/kg Se-NPs 2 بود. میزان غذادهی به ماهی­ها در طی دوره پرورشی بر اساس وزن و درجه‌حرارت آب 3-2% زی‌توده بود و روزانه در سه نوبت (ساعات 8، 15 و 21) و به مدت 8 هفته انجام شد. در پایان آزمایـش از بافت روده شـش ماهی از هر تیمار نمونه‌برداری شد. نمونه­ها طبـق روش استاندارد بافت­شناسی با استفاده از دستگاه عمل­آوری بافت، آبگیری، شفاف‌سازی، پارافینه و قالب­گیری شدند. سپس توسط دستگاه میکروتوم برش داده شد و با روش هماتوکسیلین-ائوزین (H&E) رنگ‌آمیزی و توسط میکروسکوپ نوری بررسی شدند. آسیب­های مشاهده شده در بافت روده شامل استحاله (دژنراسیون) پرزها، نفوذ یاخته های التهابی در لایه زیر مخاطی، نفوذ یاخته های التهابی در لایه عضلانی، پرخونی (زیر لایه سروزی) و نکروز در لایه عضلانی بود. در مجموع کمترین میزان آسیب در تیمار 5 (برابر با شاهد) و سپس تیمار 8 مشاهده شد. نتایج نشان می‌دهد که استفاده تلفیقی از نانوذرات آهن و سلنیوم به ترتیب با دوزهای 100 میلی‌گرم نانوذره آهن و 1 میلی‌گرم نانوذره سلنیوم در هر کیلوگرم جیره غذایی فیل‌ماهی مناسب‌تر است.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Histopathology of dietary exposure to selenium and iron nanoparticles on the intestine in reared beluga, Huso huso

نویسندگان [English]

  • Forouzan Bagherzadeh Lakani
  • Soheil Bazari Moghaddam,
  • Mehdi Masoumzadeh
  • Ayoub Yousefi Jourdehi
  • ,Jalil Jalilpoor
International Sturgeon Research Institute, Iranian Fisheries Science Research Institute, Agricultural Research Education and Extension Organization (AREEO), Rasht, Guilan, Iran
چکیده [English]

Introduction: Sustainable aquaculture relies on balanced diets that ensure optimal growth and health. Nanotechnology applications in aquaculture, such as using selenium and iron nanoparticles (Se-NPs, Fe-NPs), show promise for enhancing nutrition. However, their combined effects on intestinal health in beluga are unclear. Therefore, this study aimed to investigate the individual and combined effects of different dietary levels of Se-NPs and Fe-NPs on the intestinal histopathology of juvenile beluga, to identify the optimal and safest supplementation ratio.
Materials and methods: A total of 135 beluga juveniles (with an initial weight of 276.4 ± 32.3 g and an initial length of 40 ± 2 cm) were randomly distributed in 27 fiberglass tanks at 9 different treatments with 3 replicates. Treatments included T1 (control, no added NPs), T2 (1 mg/kg Se-NPs), T3 (2 mg/kg Se-NPs), T4 (100 mg/kg Fe-NPs), T5 (100 mg/kg Fe-NPs +1 mg/kg Se-NPs), T6 (100 mg/kg Fe-NPs +2 mg/kg Se-NPs), T7 (200 mg/kg Fe-NPs), T8 (200 mg/kg Fe-NPs +1 mg/kg Se-NPs), and T9 (200 mg/kg Fe-NPs +2 mg/kg Se-NPs). Before starting the experiment, the fish were adapted to the rearing conditions for a week. The amount of feed during the rearing period was 2-3% of the fish's weight, adjusted according to water temperature. The fish were fed three times a day (8:00-15:00 and 21:00) for 8 weeks. To find the histopathological changes, six fish intestine tissues from each group (two fish of each replicate) were sampled at the end of the experiment. Samples were dehydrated by routine methods and embedded in paraffin wax. They were sectioned by microtome and stained with Hematoxylin and Eosin (H&E).
Results and discussion: Histopathological examination revealed intestinal alterations including villi degeneration, inflammatory cell infiltration in both submucosal and muscular layers, hyperemia, and necrosis. Evaluation of treatment responses showed that the combination of 100 mg/kg Fe-NPs with 1 mg/kg Se-NPs (T₅) demonstrated the most favorable outcome, with tissue integrity comparable to the control group. The T₈ treatment also showed relatively preserved intestinal morphology. In contrast, treatments with higher nanoparticle concentrations exhibited significantly more severe pathological changes, including prominent hyperemia and inflammatory infiltration. These findings indicate that the 100:1 Fe-NPs to Se-NPs ratio effectively mitigates the adverse effects of both elements.
Conclusions: It is concluded that the combined use of iron and selenium nanoparticles at doses of 100 mg/kg Fe-NPs and 1 mg/kg Se-NPs in the diet of beluga is the most appropriate and optimal combination, resulting in minimal histopathological damage to the intestinal tissue. This ratio effectively balances the pro-oxidant potential of iron with the antioxidant capacity of selenium, thereby preserving intestinal health and integrity.
Conflicts of interest: The authors have no conflicts of interest to declare for the publication of the present work.
Funding: This article is a part of the project number 12-32-12-011-98001-991148, which was carried out with the agricultural research education and extension organization (AREEO).
Acknowledgments: The authors are grateful to researchers of the International Sturgeon Research Institute for their support during this study.

کلیدواژه‌ها [English]

  • Beluga (Huso huso)
  • Nanoparticle
  • Selenium
  • Iron
  • Histopathology
  • Intestine

مراجع

Abd El-Kader, M.F., Fath El-Bab, A.F., Abd-Elghany, M.F., Abdel-Warith, A.W.A., Younis, E.M., Dawood, M.A. 2021. Selenium nanoparticles act potentially on the growth performance, hemato-biochemical indices, antioxidative, and immune-related genes of European sea bass (Dicentrarchus labrax). Biological Trace Element Research 199: 3126-3134. DOI: 10. 1007/ s12011-​020-​02431-1
Abdel Tawwab, M., Samir, F., Abd El-Naby, A.S., Monier, M.N. 2018. Antioxidative and immunostimulatory effect of dietary cinnamon nanoparticles on the performance of Nile tilapia, Oreochromis niloticus (L.) and its susceptibility to hypoxia stress and Aeromonas hydrophila infection. Fish & Shellfish Immunology 74: 19-25. DOI: 10.1016/j.fsi.2017.12.033.
Abdel-Tawwab, M., Shukry, M., Farrag, F.A., El-Shafai, N.M., Dawood, M.A., Abdel-Latif, H.M. 2021. Dietary sodium butyrate nanoparticles enhanced growth, digestive enzyme activities, intestinal histomorphometry, and transcription of growth-related genes in Nile tilapia juveniles. Aquaculture 536: 736467. DOI: 10.1016/j.aquaculture.2021.736467.
Antony Jesu Prabhu, P., Schrama, J.W., Kaushik, S.J. 2016. Mineral requirements of fish: a systematic review. Reviews in Aquaculture 8: 172-219. DOI: 10.1111/raq.12090.
Behera, T., Swain, P., Rangacharulu, P.V., Samanta, M. 2014. Nano-Fe as feed additive improves the hematological and immunological parameters of fish Labeo rohita H. Applied Nanoscience 6: 687-694. DOI: 10.1007/s13204-013-0251-8.
Bhattacharyya, A., Reddy, S.J., Hasan, M.M., Adeyemi, M.M., Marye, R.R., Naika, H. 2015. Nanotechnology-a unique future technology in aquaculture for the food security. International Journal of Bioassays 4: 4115-4126.
Brigelius-Flohé, R., Maiorino, M. 2013. Glutathione peroxidases. Biochimica et Biophysica Acta (BBA)-General Subjects 1830: 3289-3303. DOI: 10.1016/j.bbagen.2012.11.020.
Chris, U.O., Singh, N., Agarwal, A. 2018. Nanoparticles as feed supplement on growth behaviour of cultured catfish (Clarias gariepinus) fingerlings. Materials Today: Proceedings 5: 9076-9081. DOI: 10.1016/j.matpr.2017.10.023.
Dawood, M.A., Zommara, M., Eweedah, N.M., Helal, A.I. 2020a. The evaluation of growth performance, blood health, oxidative status and immune-related gene expression in Nile tilapia (Oreochromis niloticus) fed dietary nanoselenium spheres produced by lactic acid bacteria. Aquaculture 515: 734571. DOI: 10. 1016/j. Aquaculture. 734571.
Dawood, M.A., Eweedah, N.M., Moustafa, E.M., El-Sharawy, M.E., Soliman, A.A., Amer, A.A., Atia, M.H. 2020b. Copper nanoparticles mitigate the growth, immunity, and oxidation resistance in common carp (Cyprinus carpio). Biological Trace Element Research 198: 283-292. DOI: 10. 1007/ s12011-​020-​02068-0.
Dawood, M.A., Zommara, M., Eweedah, N.M., Helal, A.I. 2020c. Synergistic effects of selenium nanoparticles and vitamin E on growth, immune-related gene expression, and regulation of antioxidant status of Nile tilapia (Oreochromis niloticus). Biological Trace Element Research 195: 624-635. DOI: 10.1007/s12011-019-01857-6.
El-Sharawy, M.E., Hamouda, M., Soliman, A.A., Amer, A.A., El-Zayat, A.M., Sewilam, H., Younis, E.M., Abdel-Warith, A.W.A., Dawood, M.A. 2021. Selenium nanoparticles are required for the optimum growth behavior, antioxidative capacity, and liver wellbeing of Striped catfish (Pangasianodon hypophthalmus). Saudi Journal of Biological Sciences 28: 7241-7247. DOI: 10.1016/j.sjbs.2021.08.023.
Ghaniem, S., Nassef, E., Zaineldin, A.I., Bakr, A., Hegazi, S. 2022. A comparison of the beneficial effects of inorganic, organic, and elemental nano-selenium on Nile tilapia: growth, immunity, oxidative status, gut morphology, and immune gene expression. Biological Trace Element Research 200: 5226-5241. DOI: 10.1007/s12011-021-03075-5.
Ghazi, S., Diab, A. M., Khalafalla, M. M., Mohamed, R. A. 2022. Synergistic effects of selenium and zinc oxide nanoparticles on growth performance, hemato-biochemical profile, immune and oxidative stress responses, and intestinal morphometry of Nile tilapia (Oreochromis niloticus). Biological Trace Element Research 200: 364-374. DOI: 10.1007/s12011-021-02631-3.
Halliwell, B., Gutteridge, J.M. 2015. Free radicals in biology and medicine. Oxford University Press, Oxford, New York, 823 p.
Hung, S.S., Groff, J.M., Lutes, P.B., Fynn-Aikins, F.K. 1990. Hepatic and intestinal histology of juvenile white sturgeon fed different carbohydrates. Aquaculture 87: 349-360. DOI: 10.1016/0044-8486(90)90072-U
Ibrahim, M.S., El‐gendy, G.M., Ahmed, A.I., Elharoun, E.R., Hassaan, M.S. 2021. Nanoselenium versus bulk selenium as a dietary supplement: Effects on growth, feed efficiency, intestinal histology, haemato‐biochemical and oxidative stress biomarkers in Nile tilapia (Oreochromis niloticus Linnaeus, 1758) fingerlings. Aquaculture Research 52: 5642-5655. DOI: 10.1111/are.15439.
Iheanacho, S.C., Ikwo, N., Igweze NO, Chukwuidha C, Ogueji EO, Onyeneke R. 2018. Effect of different dietary inclusion levels of melon seed (Citrullus lanatus) peel on growth, haematology and histology of Oroechromis niloticus juvenile. Turkish Journal of Fisheries and Aquatic Sciences 18: 377-384. DOI: 10.4194/1303-2712-v18_3_03.
Kazemi E, Sourinejad I, Ghaedi A, Johari SA, Ghasemi Z. 2020. Effect of different dietary zinc sources (mineral, nanoparticulate, and organic) on quantitative and qualitative semen attributes of rainbow trout (Oncorhynchus mykiss). Aquaculture 515: 734529. DOI: 10.1016/j.aquaculture.2019.734529.
Khalil, H.S., Maulu, S., Verdegem, M., Abdel‐Tawwab, M. 2023. Embracing nanotechnology for selenium application in aquafeeds. Reviews in Aquaculture 15: 112-129. DOI: 10.1111/raq.12705.
Khosravi-Katuli, K., Prato, E., Lofrano, G., Guida, M., Vale, G., Libralato, G. 2017. Effects of nanoparticles in species of aquaculture interest. Environmental Science and Pollution Research 24: 17326-17346. DOI: 10.1007/s11356-017-9360-3.
Kord, M.I., Srour, T.M., Farag, A.A., Omar, E.A., Nour, A.M., Khalil, H.S. 2021. The immunostimulatory effects of commercial feed additives on growth performance, non-specific immune response, antioxidants assay, and intestinal morphometry of Nile tilapia, Oreochromis niloticus. Frontiers in Physiology 12: 111. DOI: 10.3389/fphys.2021.627499.
Lall, S.P. 2002. The minerals. In: Hardy, R.H., Halver, J.E. (Eds.). Fish Nutrition, 3rd edn. Academic Press, San Diego, CA, USA, 259-308.
Mohammady, E.Y., Elashry, M.A., Ibrahim, M.S., Elarian, M., Salem, S.M., El-Haroun, E.R., Hassaan, M.S. 2024. Nano iron versus bulk iron forms as functional feed additives: growth, body indices, hematological assay, plasma metabolites, immune, anti-oxidative ability, and intestinal morphometric measurements of Nile tilapia, Oreochromis niloticus. Biological Trace Element Research 202: 787-799. DOI: 10.1007/s12011-023-03708-x.
Nair, M., Jayant, R.D., Kaushik, A., Sagar, V. 2016. Getting into the brain: Potential of nanotechnology in the management of NeuroAIDS. Advanced Drug Delivery Reviews 103: 202-217. DOI: 10.1016/j.addr.2016.02.008.
Nasr-Eldahan, S., Nabil-Adam, A., Shreadah, M.A., Maher, A.M., Ali, TE-S. 2021. A review article on nanotechnology in aquaculture sustainability as a novel tool in fish disease control. Aquaculture International 29: 1-22. DOI: 10.1007/s10499-021-00677-7.
National Research Council (NRC). 2011. Nutrient requirements of fish and shrimp. National Academy Press. Washington: DC., 392 p.
 Stankus, A. 2021. State of world aquaculture 2020 and regional reviews: FAO webinar series. FAO Aquaculture Newsletter 63: 17-18.
Ogunji, J.O., Nimptsch, J., Wiegand, C., Schulz, C., Rennert, B. 2011. Effect of housefly maggot meal (magmeal) diets on catalase, and glutathione S-transferase in the liver and gills of carp Cyprinus carpio fingerling. International Aquatic Research 3: 11-20.
Papanikolaou, G., Pantopoulos, K. 2005. Iron metabolism and toxicity. Toxicology and Applied Pharmacology 202: 199-211.  DOI: 10.1016/j.taap.2004.06.021.
Ramsden, S.R., Smith, T.J., Shaw, B.J. Handy, R.D. 2009. Dietary exposure to titanium dioxide nanoparticles in rainbow trout, (Oncorhynchus mykiss): No effect on growth, but subtle biochemical disturbances in the brain. Ecotoxicology 18: 939-951. DOI: 10.1007/s10646-009-0357-7.
Saffari, S., Keyvanshokooh, S., Zakeri, M., Johari, S.A., Pasha-Zanoosi, H., Mozanzadeh, M.T. 2018. Effects of dietary organic, inorganic, and nanoparticulate selenium sources on growth, hemato-immunological, and serum biochemical parameters of common carp (Cyprinus carpio). Fish Physiology and Biochemistry 44: 1087-1097. DOI: 10. 1007/s10695-​018-​0496-y.
Schwaiger, J., Wanke, R., Adam, S., Pawert, M., Honnen, W., Triebskorn, R. 1997. The use of histopathological indicators to evaluate contaminant-related stress in fish. Journal of Aquatic Ecosystem Stress and Recovery 6: 75-86.
Shenkin, A. 2006. Micronutrients in health and disease. Postgraduate Medical Journal 82: 559-567. DOI: 10.1136/pgmj.2006.047670.
Shi, J., Votruba, A.R., Farokhzad, O.C., Langer, R. 2010. Nanotechnology in drug delivery and tissue engineering: from discovery to applications. Nano Letters 10: 3223-30. DOI: 10.1021/nl102184c.
Terova, G., Rimoldi, S., Izquierdo, M., Pirrone, C., Ghrab, W., Bernardini, G. 2018. Nano-delivery of trace minerals for marine fish larvae: influence on skeletal ossification, and the expression of genes involved in intestinal transport of minerals, osteoblast differentiation, and oxidative stress response. Fish Physiology and Biochemistry 44: 1375-1391. DOI: 10.1007/s10695-018-0528-7.
Wang, H., Zhu, H., Wang, X., Li, E., Du, Z., Qin, J., Chen, L. 2018.  Comparison of copper bioavailability in copper-methionine, nano-copper oxide and copper sulfate additives in the diet of Russian sturgeon Acipenser gueldenstaedtii. Aquaculture 482: 146-154. DOI: 10.1016/j.aquaculture.2017.09.037.
Wang, Y., Yan, X., Fu, I. 2013. Effect of selenium nanoparticles with different sizes in primary cultured intestinal epithelial cells of crucian carp, Carassius auratus gibelio. International Journal of Nano Medicine 8: 4007-4013. DOI: 10.2147/IJN.S43691.
Watanabe, T., Kiron, V., Satoh, S. 1997. Trace mineral in fish nutrition. Aquaculture 151: 185-207. DOI: 10.1016/S0044-8486(96)01503-7.

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