Advertisement

Effects of dietary thyme essential oil on hemato-immunological indices, intestinal morphology, and microbiota of Nile tilapia

  • Gustavo Moraes Ramos Valladão
  • Sílvia Umeda Gallani
  • Suzana Kotzent
  • Inácio Mateus Assane
  • Fabiana PilarskiEmail author
Article
  • 35 Downloads

Abstract

Thyme (Thymus vulgaris) essential oil (TVEO) is a herbal medicine with one of the highest levels of antimicrobial activity. Although TVEO has been broadly used in poultry and swine production due to its immunostimulatory and growth-promoting characteristics, the effects of TVEO on fish are poorly characterized. In this study, Nile tilapia (Oreochromis niloticus L.) were fed 0, 0.1, 0.5, and 1% TVEO for 15 days. Subsequently, blood parameters, intestinal morphology, and the population of Bacillus bacteria in the intestine were evaluated. The numbers of lymphocytes (p < 0.05) and leukocytes (p < 0.05) significantly increased in the blood of the fish fed the highest dose of TVEO. Based on the normal behavior of the fish and the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, which were not altered (p > 0.05), this study concluded that the diets were safe and showed no negative or toxic effects. Even at doses as high as 1%, TVEO did not alter the population of beneficial Bacillus bacteria in the gut. In conclusion, supplementation with TVEO stimulated the cellular components of the non-specific immune response of Nile tilapia without causing deleterious effects or altering the population of important intestinal bacteria.

Keywords

Bacillus Herbal medicine Immunostimulant Innate immunity Plant extract 

Notes

Acknowledgments

The authors thank Dra. Vany P. Ferraz (Chromatography Laboratory, Department of Chemistry, UFMG) for the assistance with the analysis of the EOs. The manuscript was edited for proper English language, grammar, punctuation, spelling, and overall style by one or more of the highly qualified native English speaking editors at American Journal Experts.

Funding information

This work was supported by grants from the National Council for Scientific and Technological Development-CNPq (140487/2014-0), and the São Paulo Research Foundation-FAPESP and the Coordination of Superior Level Staff Improvement-CAPES (2014/14039-9).

Compliance with ethical standards

The experiment was approved by the Ethics Committee on the Use of Animals (CEUA) of the School of Agricultural Sciences and Veterinary Medicine at Universidade Estadual Paulista (UNESP), Jaboticabal, SP, Brazil, under protocol number 13019/15. The approval was consistent with the ethical principles adopted by the Brazilian College of Animal Experimentation.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Abdollahzadeh E, Rezaei M, Hosseini H (2014) Antibacterial activity of plant essential oils and extracts: the role of thyme essential oil, nisin, and their combination to control Listeria monocytogenes inoculated in minced fish meat. Food Control 35:177–183.  https://doi.org/10.1016/j.foodcont.2013.07.004 CrossRefGoogle Scholar
  2. Al-Sagheer AA, Mahmoud HK, Reda FM, Mahgoub SA, Ayyat MS (2017) Supplementation of diets for Oreochromis niloticus with essential oil extracts from lemongrass (Cymbopogon citratus) and geranium (Pelargonium graveolens) and effects on growth, intestinal microbiota, antioxidant and immune activities. Aquac Nutr 24:1006–1014.  https://doi.org/10.1111/anu.12637 CrossRefGoogle Scholar
  3. Ambrosio CM, Alencar SM, Sousa RL, Moreno AM, Gloria EM (2017) Antimicrobial activity of several essential oils on pathogenic and beneficial bacteria. Ind Crop Prod 97:128–136.  https://doi.org/10.1016/j.indcrop.2016.11.045 CrossRefGoogle Scholar
  4. AOAC (2006) Official methods of analysis, 18th edn. Association of Official Analytical Chemists, GaithersburgGoogle Scholar
  5. Biller-Takahashi JD, Takahashi LS, Saita MV, Gimbo RY, Urbinati EC (2013) Leukocytes respiratory burst activity as indicator of innate immunity of pacu Piaractus mesopotamicus. Braz J Biol 73:425–429.  https://doi.org/10.1590/S1519-69842013000200026 CrossRefPubMedGoogle Scholar
  6. Dauqan EM, Abdullah A (2017) Medicinal and functional values of thyme (Thymus vulgaris L.) herb. J Appl Biol Biotechnol 5:17–22.  https://doi.org/10.7324/JABB.2017.50203 CrossRefGoogle Scholar
  7. Ellis AE (1990) Lysozyme Assays. In: Stolen JS, Fletcher TC, Anderson DP, Roberson BS, Muiswinkel WB (eds) Techniques in fish immunology. Sos Publications, California, pp 101–103Google Scholar
  8. Fabio A, Cermelli C, Fabio G, Nicoletti P, Quaglio P (2007) Screening of the antibacterial effects of a variety of essential oils on microorganisms responsible for respiratory infections. Phytother Res 21:374–377.  https://doi.org/10.1002/ptr.1968 CrossRefPubMedGoogle Scholar
  9. FAO (2016) The state of world fisheries and aquaculture 2016. Contributing to food security and nutrition for all. Rome, p 200Google Scholar
  10. Gundersen H J G (1977) Notes on the estimation of the numerical density of arbitrary profiles: the edge effect. J Microsc 111:219–223Google Scholar
  11. Gutierrez J, Rodriguez G, Barry-Ryan C, Bourke P (2008) Efficacy of plant essential oils against foodborne pathogens and spoilage bacteria associated with ready-to-eat vegetables: antimicrobial and sensory screening. J Food Prot 71:1846–1854.  https://doi.org/10.4315/0362-028X-71.9.1846 CrossRefPubMedGoogle Scholar
  12. Han GQ, Xiang ZT, Yu B, Chen DW, Qi HW, Mao XB, Chen H, Mao Q, Huang ZQ (2012) Effects of different starch sources on Bacillus spp. in intestinal tract and expression of intestinal development related genes of weanling piglets. Mol Biol Rep 39:1869–1876.  https://doi.org/10.1007/s11033-011-0932-x CrossRefPubMedGoogle Scholar
  13. Haygood AM, Jha R (2016) Strategies to modulate the intestinal microbiota of Tilapia (Oreochromis sp.) in aquaculture: a review. Rev Aquac 10:320–333.  https://doi.org/10.1111/raq.12162 CrossRefGoogle Scholar
  14. Hernández A, García BG, Caballero MJ, Hernández MD (2016) The inclusion of thyme essential oil in the feed of gilthead seabream (Sparus aurata) promotes changes in the frequency of lymphocyte aggregates in gut-associated lymphoid tissue. Aquac Res 47:3341–3345.  https://doi.org/10.1111/are.12758 CrossRefGoogle Scholar
  15. Khan RU, Naz S, Nikousefat Z, Tufarelli V, Laudadio V (2012) Thymus vulgaris: alternative to antibiotics in poultry feed. World's Poult Sci J 68:401–408.  https://doi.org/10.1017/S0043933912000517 CrossRefGoogle Scholar
  16. Navarrete P, Toledo I, Mardones P, Opazo R, Espejo R, Romero J (2010) Effect of Thymus vulgaris essential oil on intestinal bacterial microbiota of rainbow trout, Oncorhynchus mykiss (Walbaum) and bacterial isolates. Aquac Res 41:667–678.  https://doi.org/10.1111/j.1365-2109.2010.02590.x CrossRefGoogle Scholar
  17. Olmos J, Paniagua-Michel J (2014) Bacillus subtilis a potential probiotic bacterium to formulate functional feeds for aquaculture. J Microb Biochem Technol 6:361–365.  https://doi.org/10.4172/1948-5948.1000169 CrossRefGoogle Scholar
  18. Osorio A, Silva TM, Duarte LP, Ferraz VP, Pereira MT, Mercadante-Simões MO, Evangelista FCG, Sabino AP, Alcântara AF (2015) Essential oil from flowers of Solanum stipulaceum: composition, effects of γ-radiation, and antileukemic activity. J Braz Chem Soc 26:2233–2240.  https://doi.org/10.5935/0103-5053.20150209. CrossRefGoogle Scholar
  19. Ouwehand AC, Tiihonen K, Kettunen H, Peuranen S, Schulze H, Rautonen N (2010) In vitro effects of essential oils on potential pathogens and beneficial members of the normal microbiota. Vet Med 55:71–78CrossRefGoogle Scholar
  20. Pfaffl MW (2004) Quantification strategies in real-time PCR. In: Bustin SA (ed) A-Z of quantitative PCR. International University Line, California, pp 87–112Google Scholar
  21. Placha I, Takacova J, Ryzner M, Cobanova K, Laukova A, Strompfova V, Venglovska K, Faix S (2014) Effect of thyme essential oil and selenium on intestine integrity and antioxidant status of broilers. Br Poult Sci 55:105–114.  https://doi.org/10.1080/00071668.2013.873772 CrossRefPubMedGoogle Scholar
  22. Ranzani-Paiva MJT, Pádua SB, Tavares-Dias M, Egami MI (2013) Métodos para análise hematológica em peixes. EDUEM, MaringáCrossRefGoogle Scholar
  23. Raut JS, Karuppayil SM (2014) A status review on the medicinal properties of essential oils. Ind Crop Prod 62:250–264.  https://doi.org/10.1016/j.indcrop.2014.05.055 CrossRefGoogle Scholar
  24. Ren P, Xu L, Yang Y, He S, Liu W, Ringø E, Zhou Z (2013) Lactobacillus planarum subsp. plantarum JCM 1149 vs. Aeromonas hydrophila NJ-1 in the anterior intestine and posterior intestine of hybrid tilapia Oreochromis niloticus♀× Oreochromis aureus♂: an ex vivo study. Fish Shellfish Immunol 35:146–153.  https://doi.org/10.1016/j.fsi.2013.04.023 CrossRefPubMedGoogle Scholar
  25. Rieger AM, Barreda DR (2011) Antimicrobial mechanisms of fish leukocytes. Dev Comp Immunol 35:1238–1245.  https://doi.org/10.1016/j.dci.2011.03.009 CrossRefPubMedGoogle Scholar
  26. Saleh N, Allam T, El-latif AA, Ghazy E (2014) The effects of dietary supplementation of different levels of thyme (Thymus vulgaris) and ginger (Zingiber officinale) essential oils on performance, hematological, biochemical and immunological parameters of broiler chickens. Glob Vet 12:736–744.  https://doi.org/10.5829/idosi.gv.2014.12.06.83189 CrossRefGoogle Scholar
  27. Santos WM, Brito TS, Prado SDA, Oliveira CG, Paula AC, Melo DC, Ribeiro PA (2016) Cinnamon (Cinnamomum sp.) inclusion in diets for Nile tilapia submitted to acute hypoxic stress. Fish Shellfish Immunol 54:551–555.  https://doi.org/10.1016/j.fsi.2016.04.135 CrossRefGoogle Scholar
  28. Scapigliati G (2013) Functional aspects of fish lymphocytes. Dev Comp Immunol 41:200–208.  https://doi.org/10.1016/j.dci.2013.05.012 CrossRefPubMedGoogle Scholar
  29. Song SK, Beck BR, Kim D, Park J, Kim J, Kim HD, Ringø E (2014) Prebiotics as immunostimulants in aquaculture: a review. Fish Shellfish Immunol 40:40–48.  https://doi.org/10.1016/j.fsi.2014.06.016 CrossRefPubMedGoogle Scholar
  30. Sönmez AY, Bilen S, Alak G, Hisar O, Yanık T, Biswas G (2015) Growth performance and antioxidant enzyme activities in rainbow trout (Oncorhynchus mykiss) juveniles fed diets supplemented with sage, mint and thyme oils. Fish Physiol Biochem 41:165–175.  https://doi.org/10.1007/s10695-014-0014-9 CrossRefPubMedGoogle Scholar
  31. Sutili FJ, Velasquez A, Pinheiro CG, Heinzmann BM, Gatlin DM, Baldisserotto B (2016) Evaluation of Ocimum americanum essential oil as an additive in red drum (Sciaenops ocellatus) diets. Fish Shellfish Immunol 56:155–161.  https://doi.org/10.1016/j.fsi.2016.07.008 CrossRefPubMedGoogle Scholar
  32. Sutili FJ, Gatlin DM, Heinzmann BM, Baldisserotto B (2017) Plant essential oils as fish diet additives: benefits on fish health and stability in feed. Rev Aquacult 10:716–726.  https://doi.org/10.1111/raq.12197 CrossRefGoogle Scholar
  33. Tran NT, Xiong F, Hao YT, Zhang J, Wu SG, Wang GT (2017) Two biomass preparation methods provide insights into studying microbial communities of intestinal mucosa in grass carp (Ctenopharyngodon idellus). Aquac Res 48:4272–4283.  https://doi.org/10.1111/are.13248 CrossRefGoogle Scholar
  34. Valladão GMR, Gallani SU, Pilarski F (2015) Phytotherapy as an alternative for treating fish disease. J Vet Pharmacol Ther 38:417–428.  https://doi.org/10.1111/jvp.12202 CrossRefPubMedGoogle Scholar
  35. Valladão GMR, Gallani SU, Pala G, Jesus RB, Kotzent S, Costa JC, Silva TFA, Pilarski F (2017) Practical diets with essential oils of plants activate the complement system and alter the intestinal morphology of Nile tilapia. Aquac Res 48:5640–5649.  https://doi.org/10.1111/are.13386 CrossRefGoogle Scholar
  36. Vallejos-Vidal E, Reyes-López F, Teles M, MacKenzie S (2016) The response of fish to immunostimulant diets. Fish Shellfish Immunol 56:34–69.  https://doi.org/10.1016/j.fsi.2016.06.028 CrossRefPubMedGoogle Scholar
  37. Van Hai N (2015) Research findings from the use of probiotics in tilapia aquaculture: a review. Fish Shellfish Immunol 45:592–597.  https://doi.org/10.1016/j.fsi.2015.05.026 CrossRefPubMedGoogle Scholar
  38. Zadmajid V, Mohammadi CH (2017) Dietary thyme essential oil (Thymus vulgaris) changes serum stress markers, enzyme activity, and hematological parameters in gibel carp (Carassius auratus gibelio) exposed to silver nanoparticles. Iran J Fish Sci 16:1063–1084Google Scholar
  39. Zanuzzo FS, Urbinati EC, Rise ML, Hall JR, Nash GW, Gamperl AK (2015) Aeromonas salmonicida induced immune gene expression in Aloe vera fed steelhead trout, Oncorhynchus mykiss (Walbaum). Aquaculture 435:1–9.  https://doi.org/10.1016/j.aquaculture.2014.09.010 CrossRefGoogle Scholar
  40. Zeppenfeld CC, Hernández DR, Santinón JJ, Heinzmann BM, Cunha MA, Schmidt D, Baldisserotto B (2016) Essential oil of Aloysia triphylla as feed additive promotes growth of silver catfish (Rhamdia quelen). Aquac Nutr 22:933–940.  https://doi.org/10.1111/anu.12311 CrossRefGoogle Scholar
  41. Zorriehzahra MJ, Delshad ST, Adel M, Tiwari R, Karthik K, Dhama K, Lazado CC (2016) Probiotics as beneficial microbes in aquaculture: an update on their multiple modes of action: a review. Vet Q 36:228–241.  https://doi.org/10.1080/01652176.2016.1172132 CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Postgraduate Program in AquacultureUniversidade Nilton LinsManausBrazil
  2. 2.Microbiology and Parasitology LaboratoryAquaculture Center at Universidade Estadual Paulista (CAUNESP)JaboticabalBrazil
  3. 3.Department of Agricultural MicrobiologyUniversidade Estadual Paulista (UNESP)JaboticabalBrazil

Personalised recommendations