Comparative Epigenomic Profiling and Gene Expression Patterns of Zebrafish, Danio rerio, Administrated by Dietary Agrimos®

Document Type : Original Research

Authors
1 Department of Natural Resources, Isfahan University of Technology, Isfahan, P.O. Box: 84156-83111, Islamic Republic of Iran.
2 Faculty of Natural Resources, University of Tehran, Karaj, Islamic Republic of Iran.
Abstract
The different aspects of using dietary supplements such as prebiotics in aquaculture and their effects on innate immune response, and especially their vertical transmission, are of a grave importance. To address such issues in both horizontal and vertical transmission of boosting immune system, the present study was designed to investigate the effect of different levels of dietary Agrimos® on the innate immune-related gene expression [Lysozyme (Lyz) and Tumor Necrosis Factor alpha (TNFα)], DNA methylation, and three Histone MethylTransferase [HMTs (H3K4, H3K9, H3K27)] activities as well as growth performance in zebrafish (Danio rerio). Three hundred and sixty healthy 24-days-old zebrafish were randomly distributed in twelve aquaria assigned to four groups. Zebrafish were fed with either control diet or a diet supplemented by different levels (0.2, 0.4, and 0.8%) of Agrimos® for 90 days. The offspring of each treatment was assessed to find the potential of vertical transmission of immunity by using this dietary prebiotic. At the end of the experiment, gene expression studies revealed significant up-regulation (P˂ 0.05) of TNFα and Lyz genes in 0.2 and 0.4% Agrimos® fed fish compared with the control group. Although our findings showed that supplemented diet reduced DNA methylation (P˂ 0.05) in Agrimos® treatments compared with the control, there was no significant change in all three HMTs’ activities among experimental groups (P> 0.05). The result shows the successful transmission of Lyz gene expression as an innate immune response to the offspring of the treated adults and supports a direct role of DNA demethylation in the regulation of these candidate gene expressions, suggesting possible role of diet on regulating the epigenetic processes.

Keywords


1. Akbari, M., Taghizadeh, V., Heidarieh, M. and Hajimoradloo, A. 2017. The key role of tumor necrosis factor alpha (TNF-α) in vaccinated rainbow trout via irradiated Ichthyophthirius multifiliis trophont. Vet Arh, 87(2), 229-237.
2. Akhter, N., Wu, B., Memon, A. M. and Mohsin, M. 2015. Probiotics and prebiotics associated with aquaculture: A review. Fish Shellfish Immun, 45, 733-741.
3. Anderson, I., Reiner, A. H., Aanes, H., Alestrom, P. and Collas, P. 2012. Developmental features of DNA methylation during activation of the embryonic zebrafish genome. Genome Biol, 13, 65-78.
4. Beikzadeh, M., Peighambardoust, S.H., Beikzadeh, S. and Homayoun-Rad, A. 2017. Effects of Inulin, Oligofroctose and Oligofroctose-enriched Inulin on physiochemical, staling and sensory properties of prebiotic cake. J Agr Sci Tech, 19, 1241-1252.
5. Buentello, J. A., Neill, W. H. and Gatlin, D. M. 2010. Effects of dietary prebiotics on the growth, feed efficiency and non-specific immunity of juvenile red drum Sciaenops ocellatus fed soybean-based diets. Aquac Res, 41, 411-418.
6. Cerezuela, R., Meseguer, J. and Esteban, A. 2011. Current knowledge in synbiotic use for fish aquaculture: a review. J Aquac Res Dev. https://doi.org/10.4172/2155-9546, S1-008
7. Chernyavskaya, Y., Mudbhary, R., Zhang, C., Tokarz, D., Jacob, V., Gopinath, S., Sun, X. and Wang, s. 2017. Loss of DNA methylation in zebrafish embryos activates retrotransposons to trigger antiviral signaling. Development, 144, 2925-2939.
8. Davidovic, R. S., Bozovic, A. M., Mandusic, V. L. and Krajnovic, M. M. 2014. Methylation-specific PCR: four steps in primer design. Cent Eur J Biol, 9, 1127-1139.
9. EU. 2010. Directive 2010/63/EU of the European parliament and of the council of 22 September 2010 on the protection of animals used for scientific purposes. O J, 276, 33-79.
10. Forsatkar, M.N., Nematollahi, M.A., Rafiee, G., Farahmand, H. and Lawrence, C. 2018. Effects of the prebiotic mannan-oligosaccharide on feed deprived zebrafish: growth and reproduction. Aquac Res: 49(8), 2822-2832.
11. Ganguly, S., Paul, I. and Mukhopadhayay, S. K. 2010. Application and effectiveness of immunostimulants, probiotics, and prebiotics in aquaculture: a review. Isr J Aquac, 62, 130-138.
12. Gazzar, m., Yoza, B. K., Hu, J., Cousart, S. L. and McCall, C. E. 2007. Epigenetics silencing of tumor necrosis factor α during endotoxin tolerance. J Biol Chem, 282, 26857-26864.
13. Hoseinifar, S.H., Van Doan, H. and Ashouri, G. 2019. Galactooligosaccharide effects as prebiotic on intestinal microbiota of different fish species. Agro J, 14(3), 266-278.
14. Huidobro, C., Fernandez, A. F. and Fraga, M. F. 2013. Aging epigenetics: Causes and consequences. Mol Aspects Med, 34, 765-781.
15. Kim, D. and Austin, B. 2006. Innate immune responses in rainbow trout (Oncorhynchus mykiss, Walbaum) induced by probiotics. Fish Shellfish Immun, 21, 513-524.
16. Lindeman, L. C., Winata, C. L., Aanes, H., Mathavan, S., Alestrom, P. and Collas, P. 2010. Chromatin states of developmentally-regulated genes revealed by DNA and histone methylation patterns in Zebrafish embryos. Int J Dev Biol, 54, 803-813.
17. Livak, K. J. and Schmittgen, T. D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25, 402-408.
18. Massicotte, R., Whitelaw, E. and Angers, B. 2011. DNA methylation: a source of random variation in natural populations. Epigenetics, 6, 421-427.
19. Mehrad, B., Jafaryan, H. and Taati, M. 2012. Assessment of the effects of dietary vitamin E on growth performance and reproduction of zebrafish, Danio rerio (Pisces, Cyprinidae). Oceanogr Mar Sci, 3(1), 1-7.
20. Moghadam, H., Morkore, T. and Robinson, N. 2015. Epigenetics-potential for programming fish for aquaculture? J Mar Sci Eng, 3, 175-192.
21. Mohamed, K., Megahed, M. E. and Ali, M. A. M. 2017. Effect of dietary supplementation of Agrimos® on growth performance, feed utilization and immunological parameters of Macrobrachium rosenbergii juveniles. Aquacult Int, 25, 1441-1452.
22. Opazo, R., Valladares, L. and Romero, J. 2017. Comparison of gene expression patterns of key growth genes between different rate growths in zebrafish (Danio rerio) siblings. Lat Am J Aquat Res, 45, 766-775.
23. Paul, B., Barnes, S., Demark-Wahnefried, W., Morrow, C., Salvador, C., Skibola, C. h. and Tollefsbol, T. O. 2015. Influence of diet and the gut microbiome on epigenetic modulation in cancer and other diseases. Clin Epigenetics, 7, 112-123.
24. Peterson, B. C., Bramble, T. C. and Manning, B. B. 2010. Effects of Bio-Mos® on growth and survival of channel catfish challenged with Edwardsiella ictaluri. J World Aquacult Soc, 41, 149-155.
25. Piccolo, G., Centodocati, G., Bovera, F., Marrone, R. and Nizza, A. 2013. Effects of mannan oligosaccharide and inulin on sharpsnout seabream (Diplodus puntazzo) in the context of partial fish meal substitution by soybean meal. Ital J Anim Sci, 12, 133-138.
26. Ribas, L. and Piferrer, F. 2013. The zebrafish (Danio rerio) as a model organism, with emphasis on applications for finish aquaculture research. Review in Aquaculture, 5, 1-32.
27. Rolim, P. M. 2015. Development of prebiotic food products and health benefits. J Food Sci Technol, 35, 3-10.
28. Saleem, M., Abbas, K., Manan, M., Ijaz, H., Ahmed, B., Ali, M., Hanif, M., Farooqi, A. A. and Qadir, M. I. 2015. Epigenetic therapy for cancer. Pak J pharm Sci, 28, 1023-1032.
29. Sullivan, K. E., Reddy, A. B. M., Dietzmann, K., Suriano, R., Kocieda, V. P., Stewart, M. and Bhatia, M. 2007. Epigenetics regulation of tumor necrosis factor alpha. Mol Cell Biol, 27, 5147-5160.
30. Tarnecki, A.M., Burgos, F.A., Ray, C.L. and Arias, C.R. 2017. Fish intestinal microbiome: diversity and symbiosis unraveled by metagenomics. J Appl Microbiol, 123, 2-17.
31. Torrecillas, S., Makol, A., Caballero, M. J., Montero, D., Gines, R. and Sweetman, J. 2011. Improved feed utilization, intestinal mucus production and immune parameters in sea bass (Dicentrarchus labrax) fed mannan oligosaccharides (MOS). Aquac Nutr, 17, 223-233.
32. Torrecillas, S., Makol, A., Betancor, M. B., Montero, D., Caballero, M. J. and Sweetman, J. 2013. Enhanced intestinal epithelial barrier health status on European sea bass (Dicentrarchus labrax) fed mannan oligosaccharides. Fish Shellfish Immun, 34, 1485-1495.
33. Torrecillas, S., Montero, D. and Izquierdo, M. 2014. Improved health and growth of fish fed mannan oligosaccharides: Potential mode of action. Fish Shellfish Immun, 36, 525-544.
34. Triantaphyllopoulos, K. A., Ikonomopoulos, I. and Bannister, A. 2016. Epigenetics and inheritance of phenotype variation in livestock. Epigenetics Chromatin, 9, 31-49.
35. Ulloa, P. E., Medrano, J. F. and Feijoo, C. G. 2014. Zebrafish as animal model for aquaculture nutrition research, Front Genet, 5, 313-318.
36. Vaz, C., Wee, C. W., Lee, G. P., Ingham, P. W. and Tanavde, V. 2015. Deep sequencing of small RNA facilitates tissue and sex associated microRNA discovery in Zebrafish. BMC Genomics, 16, 950-966.
37. Wang, Zh. and Zhang, Sh. 2010. The role of lysozyme and complement in the antibacterial activity of Zebrafish (Danio rerio) egg cytosol. Fish Shellfish Immun, 29, 773-777.
38. Welker, T. L., Lim, C., Yildrim-Aksoy, M. and Klesius, P. H. 2011. Effect of short-term feeding duration of diets containing commercial whole-cell yeast or yeast subcomponents on immune function and disease resistance in channel catfish, Ictalurus punctatus. J Anim Physiol Anim Nutr, 96, 159-171.
39. Wu, S. F., Zhang, H., Hammoud, S. S., Potok, M., Nix, D. A., Jones, D. A. and Cairns, B. R. 2011. DNA methylation profiling in Zebrafish. Method cell biol, 104, 327-339.
40. Yar Ahmadi, P., Farahmand, H., Kolangi Miandare, H., Mirvaghefi, A. and Hoseinifar, S. H. 2014. The effects of dietary Immunogen® on innate immune response, immune related genes expression and disease resistance of rainbow trout (Oncorhynchus mykiss). Fish Shellfish Immun, 37, 209-214.
41. Yarahmadi, P., Kolangi Miandare, H., Farahmand, H., Mirvaghefi, A. and Hoseinifar, S. H. 2014. Dietary fermentable fiber upregulated immune related genes expression, increased innate immune response and resistance of rainbow trout (Oncorhynchus mykiss) against Aeromonas hydrophila. Fish Shellfish Immun, 41, 326-331.
42. Yousefi, S., Hoseinifar, S. H., Paknejad, H. and Hajimoradloo, A. 2018. The effects of dietary supplement of galactooligosaccharide on innate immunity, immune related genes expression and growth performance in zebrafish (Danio rerio). Fish Shellfish Immun, 73, 192-196.
43. Yuji-Sado, R., Raulino-Domanski, F., Freitas, P. F. and Baioco-Sales, F. 2015. Growth, immune status and intestinal morphology of Nile tilapia fed dietary prebiotics (mannan oligosaccharides-MOS). Latin American Journal of Aquaculture Research, 43, 944-952.
44. Zhang, N., Luo, G., Tan, H., Liu, W. and Hou, Z. 2016. Growth, digestive enzyme activity and welfare of tilapia reared in a biofloc-based system with poly-β-hydroxybutyric as a carbon source. Aquaculture, 464, 710-717.