Immunohematological parameters of rainbow trout (Oncorhynchus mykiss) fed supplemented diet with different forms of barberry root (Berberis vulgaris)

  • Sara Ramezanzadeh
  • Abdolmohamad Abedian KenariEmail author
  • Mohammad Esmaeili
Original Article


In this experiment, the effect of barberry root powder and methanol extract on the growth and immunohematological parameters of rainbow trout (Oncorhynchus mykiss) was studied. A total of 306 rainbow trout fingerlings (15.55 ± 0.76 g) were fed for 8 weeks with six experimental diets: control (C), BRP10 (10 g kg−1 of barberry root powder), BRP20 (20 g kg−1 of barberry root powder), BRE250 (250 mg kg−1 of barberry root extract), BRE500 (500 mg kg−1 of barberry root extract), and oxytetracycline OTC30 (30 g kg−1). The results indicated BRE250, BRE500, and OTC30 had significantly higher weight gain (93.04, 92.52, and 90.48 g, respectively) compared with other dietary treatments (P < 0.05). The highest levels of WBC (26.5 × 103), RBC (1.8 × 106), HB (9.47 g dl−1), MCHC (22.19 g dl−1), lymphocytes (67.67%), lysozyme (65.67), and ACH50 (139) in fish fed BRE500 diet were observed (P < 0.05). Also, inhibition zone appeared with the barberry against Aeromonas hydrophila in vitro. Totally, it is suggested that the BRE500 diet can be used as a natural stimulant of growth and immunity in the aquaculture industry.


Bacteria challenging Blood chemistry Immune system Immune stimulants Hematology Rainbow trout 





Mean corpuscular volume


Mean corpuscular hemoglobin


Mean corpuscular hemoglobin concentration


Alternative complement activity




Tryptic soy agar


White blood cell


Red blood cell







Thanks are extended to Mr. Kamali for helping us doing some analytical experiments.

Funding information

The authors wish to thank the Tarbiat Modares University (Noor, Mazandaran, Iran) for their financial support.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Abtahi B, Yousefi M, Kenari AA (2013) Influence of dietary nucleotides supplementation on growth, body composition and fatty acid profile of Beluga sturgeon juveniles (Huso huso). Aquac Res 44:254–260CrossRefGoogle Scholar
  2. Adel M, Amiri AA, Zorriehzahra J, Nematolahi A, Esteban MÁ (2015) Effects of dietary peppermint (Mentha piperita) on growth performance, chemical body composition and hematological and immune parameters of fry Caspian white fish (Rutilus frisii kutum). Fish Shellfish Immunol 45:841–847CrossRefGoogle Scholar
  3. Adel M, Pourgholam R, Zorriehzahra J, Ghiasi M (2016) Hemato–immunological and biochemical parameters, skin antibacterial activity, and survival in rainbow trout (Oncorhynchus mykiss) following the diet supplemented with Mentha piperita against Yersinia ruckeri. Fish Shellfish Immunol 55:267–273CrossRefGoogle Scholar
  4. Ahmadifar E, Sheikhzadeh N, Roshanaei K, Dargahi N, Faggio C (2019) Can dietary ginger (Zingiber officinale) alter biochemical and immunological parameters and gene expression related to growth, immunity and antioxidant system in zebrafish (Danio rerio)? Aquaculture 507:341–348CrossRefGoogle Scholar
  5. Altunoglu YC, Bilen S, Ulu F, Biswas G (2017) Immune responses to methanolic extract of black cumin (Nigella sativa) in rainbow trout (Oncorhynchus mykiss). Fish Shellfish Immunol 67:103–109CrossRefGoogle Scholar
  6. Amar EC, Kiron V, Satoh S, Okamoto N, Watanabe T (2000) Effects of dietary β-carotene on the immune response of rainbow trout Oncorhynchus mykiss. Fish Sci 66:1068–1075CrossRefGoogle Scholar
  7. Arowosegbe S, Wintola OA, Afolayan AJ (2012) Phytochemical constituents and allelopathic effect of Aloe ferox Mill. root extract on tomato. J. Med. Plants Res 6:2094–2099Google Scholar
  8. Asadi M, Mirvaghefei A, Nematollahi M, Banaee M, Ahmadi K (2012) Effects of Watercress (Nasturtium nasturtium) extract on selected immunological parameters of rainbow trout (Oncorhynchus mykiss). Open Vet J 2:32–39Google Scholar
  9. Awad E, Austin B (2010) Use of lupin, Lupinus perennis, mango, Mangifera indica, and stinging nettle, Urtica dioica, as feed additives to prevent Aeromonas hydrophila infection in rainbow trout (Oncorhynchus mykiss). J Fish Dis 33:413–420CrossRefGoogle Scholar
  10. Awad E, Awaad A (2017) Role of medicinal plants on growth performance and immune status in fish. Fish Shellfish Immunol 67:40–54CrossRefGoogle Scholar
  11. Awad E, Austin D, Lyndon A, Awaad A (2019) Possible effect of hala extract (Pandanus tectorius) on immune status, anti-tumour and resistance to Yersinia ruckeri infection in rainbow trout (Oncorhynchus mykiss). Fish Shellfish Immunol 87:620–626CrossRefGoogle Scholar
  12. Bilen S, Bulut M, Bilen AM (2011) Immunostimulant effects of Cotinus coggyria on rainbow trout (Oncorhynchus mykiss). Fish Shellfish Immunol 30:451–455CrossRefGoogle Scholar
  13. Bilen S, Altunoglu YC, Ulu F, Biswas G (2016a) Innate immune and growth promoting responses to caper (Capparis spinosa) extract in rainbow trout (Oncorhynchus mykiss). Fish Shellfish Immunol 57:206–212CrossRefGoogle Scholar
  14. Bilen S, Ünal S, Güvensoy H (2016b) Effects of oyster mushroom (Pleurotus ostreatus) and nettle (Urtica dioica) methanolic extracts on immune responses and resistance to Aeromonas hydrophila in rainbow trout (Oncorhynchus mykiss). Aquaculture 454:90–94CrossRefGoogle Scholar
  15. Blaxhall P, Daisley K (1973) Routine haematological methods for use with fish blood. J. Fish Biol 5:771–781CrossRefGoogle Scholar
  16. Bulfon C, Bongiorno T, Messina M, Volpatti D, Tibaldi E, Tulli F (2017) Effects of Panax ginseng extract in practical diets for rainbow trout (Oncorhynchus mykiss) on growth performance, immune response and resistance to Yersinia ruckeri. Aquac Res 48:2369–2379CrossRefGoogle Scholar
  17. Chakraborty SB, Horn P, Hancz C (2014) Application of phytochemicals as growth-promoters and endocrine modulators in fish culture. Rev Aquacult 6:1–19CrossRefGoogle Scholar
  18. Citarasu T (2010) Herbal biomedicines: a new opportunity for aquaculture industry. Aquac Int 18:403–414CrossRefGoogle Scholar
  19. Clerton P, Troutaud D, Verlhac V, Gabaudan J, Deschaux P (2001) Dietary vitamin E and rainbow trout (Oncorhynchus mykiss) phagocyte functions: effect on gut and on head kidney leucocytes. Fish Shellfish Immunol 11:1–13CrossRefGoogle Scholar
  20. Dawood MA, Koshio S, Esteban MÁ (2018) Beneficial roles of feed additives as immunostimulants in aquaculture: a review. Rev Aquacult 10:950–974CrossRefGoogle Scholar
  21. Declaration of Helsinki. Recommendations Guiding Physicians in Biomedical Research Involving Human Subjects, 29th World Medical Assembly, Tokyo, Japan, October 1975Google Scholar
  22. Dembitsky VM, Gloriozova TA, Poroikov VV (2015) Naturally occurring plant isoquinoline N-oxide alkaloids: their pharmacological and SAR activities. Phytomedicine 22:183–202CrossRefGoogle Scholar
  23. Diler O, Gormez O, Diler I, Metin S (2017) Effect of oregano (Origanum onites L.) essential oil on growth, lysozyme and antioxidant activity and resistance against Lactococcus garvieae in rainbow trout (Oncorhynchus mykiss). Aquacult. Nutr 23:844–851Google Scholar
  24. Ding J, Wang Z, Jiang S, Liu J, Liu L (2006) Effects of berberis extract on growth, nutrient apparent metabolic rate and digestive enzyme activity of broilers. Feed Review 6Google Scholar
  25. Direkbusarakom S (2011) Application of medicinal herbs to aquaculture in Asia. WJST 1:7–14Google Scholar
  26. Dügenci SK, Arda N, Candan A (2003) Some medicinal plants as immunostimulant for fish. J Ethnopharmacol 88:99–106CrossRefGoogle Scholar
  27. Esmaeili M, Abedian Kenari A, Rombenso A (2017a) Effects of fish meal replacement with meat and bone meal using garlic (Allium sativum) powder on growth, feeding, digestive enzymes and apparent digestibility of nutrients and fatty acids in juvenile rainbow trout (Oncorhynchus mykiss Walbaum, 1792). Aquac Nutr 23:1225–1234CrossRefGoogle Scholar
  28. Esmaeili M, Kenari AA, Rombenso A (2017b) Immunohematological status under acute ammonia stress of juvenile rainbow trout (Oncorhynchus mykiss Walbaum, 1792) fed garlic (Allium sativum) powder-supplemented meat and bone meal-based feeds. Comp Clin Path 26:853–866CrossRefGoogle Scholar
  29. Farsani MN, Hoseinifar SH, Rashidian G, Farsani HG, Ashouri G, Van Doan H (2019) Dietary effects of Coriandrum sativum extract on growth performance, physiological and innate immune responses and resistance of rainbow trout (Oncorhynchus mykiss) against Yersinia ruckeri. Fish Shellfish Immunol 91:233–240CrossRefGoogle Scholar
  30. Fatehi M, Saleh TM, Fatehi-Hassanabad Z, Farrokhfal K, Jafarzadeh M, Davodi S (2005) A pharmacological study on Berberis vulgaris fruit extract. J Ethnopharmacol 102:46–52CrossRefGoogle Scholar
  31. Food and Agriculture Organization (FAO) (2018) The State of World Fisheries and Aquaculture. FAO Fisheries and Aquaculture Department, RomeGoogle Scholar
  32. Freile M et al (2003) Antimicrobial activity of aqueous extracts and of berberine isolated from Berberis heterophylla. Fitoterapia 74:702–705CrossRefGoogle Scholar
  33. Ghareeb DA, Elwakeel EH, Khalil R, Aziz MS (2016) Investigation of the immunomodulatory effect of Berberis vulgaris on core-pulsed dendritic cell vaccine. BMC Complement Altern Med 16:325–336CrossRefGoogle Scholar
  34. Gómez-Estaca J, De Lacey AL, López-Caballero M, Gómez-Guillén M, Montero P (2010) Biodegradable gelatin–chitosan films incorporated with essential oils as antimicrobial agents for fish preservation. Food Microbiol 27:889–896CrossRefGoogle Scholar
  35. Gutfinger T (1981) Polyphenols in olive oils. J Am Oil Chem Soc 58:966–968CrossRefGoogle Scholar
  36. Haghighi M, Rohani MS (2013) The effects of powdered ginger (Zingiber officinale) on the haematological and immunological parameters of rainbow trout Oncorhynchus mykiss. JMPHTR 1:8–12Google Scholar
  37. Hosseinpour Aghaei R, Abedian Kenari A, Yazdani Sadati MA, Esmaeili M (2018) The effect of time-dependent protein restriction on growth factors, nonspecific immunity, body composition, fatty acids and amino acids in the Siberian sturgeon (Acipenser baerii). Aquac Res 49:3033–3044CrossRefGoogle Scholar
  38. Hosseinzadeh H, Ramezani M, Shafaei H, Taghiabadi E (2013) Anticonvulsant effect of Berberis integerrima L. root extracts in mice. J Acupunct Meridian Stud 6:12–17CrossRefGoogle Scholar
  39. Imenshahidi M, Hosseinzadeh H (2019) Berberine and barberry (Berberis vulgaris): a clinical review. Phytother Res 33:504–523CrossRefGoogle Scholar
  40. Jafari AA, Shohrati M, Mahmoudi R, Haj Hoseini R, Nosratpour S, Pajohi-Alamoti LA, Mohamad Latifi A (2014) Chemical composition and biological activities of Scrophularia striata extracts. Minerva Biotec 26:183–189Google Scholar
  41. Kaneda Y, Torii M, Tanaka T, Aikawa M (1991) In vitro effects of berberine sulphate on the growth and structure of Entamoeba histolytica, Giardia lamblia and Trichomonas vaginalis. Ann Trop Med Parasitol 85:417–425CrossRefGoogle Scholar
  42. Kenari AA, Sotoudeh E, Rezaei MH (2011) Dietary soybean phosphatidylcholine affects growth performance and lipolytic enzyme activity in Caspian brown trout (Salmo trutta Caspius) alevin. Aquac Res 42:655–663CrossRefGoogle Scholar
  43. Kenari AA, Mahmoudi N, Soltani M, Abediankenari S (2013) Dietary nucleotide supplements influence the growth, haemato-immunological parameters and stress responses in endangered Caspian brown trout (Salmo trutta caspius Kessler, 1877). Aquac Nutr 19:54–63CrossRefGoogle Scholar
  44. Kiasalari Z, Khalili M, Ahmadi P (2011) Effect of alcoholic extract of Berberis Vulgaris fruit on acute and chronic inflammation in male rats. JBUMS 13:28–35Google Scholar
  45. Koduru S, Grierson D, Van de Venter M, Afolayan A (2007) Anticancer activity of steroid alkaloids isolated from Solanum aculeastrum. Pharm Biol 45:613–618CrossRefGoogle Scholar
  46. Kosalec I, Gregurek B, Kremer D, Zovko M, Sanković K, Karlović K (2009) Croatian barberry (Berberis croatica Horvat): a new source of berberine—analysis and antimicrobial activity. World J Microbiol Biotechnol 25:145–150CrossRefGoogle Scholar
  47. Kumar N, Sharma J, Singh SP, Singh A, Krishna VH, Chakrabarti R (2019) Validation of growth enhancing, immunostimulatory and disease resistance properties of Achyranthes aspera in Labeo rohita fry in pond conditions. Heliyon 5:e01246CrossRefGoogle Scholar
  48. Langmead L, Rampton D (2001) Herbal treatment in gastrointestinal and liver disease benefits and dangers. Aliment Pharmacol Ther 15:1239–1252CrossRefGoogle Scholar
  49. Lazavi F, Mirmiran P, Sohrab G, Nikpayam O, Angoorani P, Hedayati M (2018) The barberry juice effects on metabolic factors and oxidative stress in patients with type 2 diabetes: a randomized clinical trial. Complement Ther Clin Pract 31:170–174CrossRefGoogle Scholar
  50. Lee J, Durst RW, Wrolstad RE (2005) Determination of total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines by the pH differential method: collaborative study. J AOAC Int 88:1269–1278Google Scholar
  51. Mahady GB, Pendland SL, Stoia A, Chadwick LR (2003) In vitro susceptibility of Helicobacter pylori to isoquinoline alkaloids from Sanguinaria canadensis and Hydrastis canadensis. Phytother Res 17:217–221CrossRefGoogle Scholar
  52. Matani Bour H, Esmaeili M, Abedian Kenari A (2018) Growth performance, muscle and liver composition, blood traits, digestibility and gut bacteria of beluga (Huso huso) juvenile fed different levels of soybean meal and lactic acid. Aquac Nutr 24:1361–1368CrossRefGoogle Scholar
  53. Minaiyan M, Ghannadi A, Mahzouni P, Jaffari-Shirazi E (2011) Comparative study of Berberis vulgaris fruit extract and berberine chloride effects on acetic acid-induced colitis in rats. Iran J Pharm Res 10:97–104Google Scholar
  54. Mo WY, Chen Z, Leung HM, Leung AOW (2017) Application of veterinary antibiotics in China’s aquaculture industry and their potential human health risks. Environ Sci Pollut Res Int 24:8978–8989CrossRefGoogle Scholar
  55. Moghanlou KS, Isfahani EN, Dorafshan S, Tukmechi A, Aramli MS (2018) Effects of dietary supplementation with Stachys lavandulifolia Vahl extract on growth performance, hemato-biochemical and innate immunity parameters of rainbow trout (Oncorhynchus mykiss). Anim Feed Sci Technol 237:98–105CrossRefGoogle Scholar
  56. National Research Council (2011a) In: Guide for the care and use of laboratory animals Washington, DC: National AcademyGoogle Scholar
  57. National Research Council (NRC) (2011b) Nutrient Requirements of Fish and Shrimp. The National Academies Press, Washington, DCGoogle Scholar
  58. Oskoii SB, Kohyani AT, Parseh A, Salati AP, Sadeghi E (2012) Effects of dietary administration of Echinacea purpurea on growth indices and biochemical and hematological indices in rainbow trout (Oncorhynchus mykiss) fingerlings. Fish Physiol Biochem 38:1029–1034CrossRefGoogle Scholar
  59. Oujifard A, Seyfabadi J, Kenari AA, Rezaei M (2012) Fish meal replacement with rice protein concentrate in a practical diet for the Pacific white shrimp, Litopenaeus vannamei Boone, 1931. Aquac Int 20:117–129CrossRefGoogle Scholar
  60. Potdar D, Hirwani R, Dhulap S (2012) Phyto-chemical and pharmacological applications of Berberis aristata. Fitoterapia 83:817–830CrossRefGoogle Scholar
  61. Pourmoghim H, Haghighi M, Rohani MS (2015) Effect of dietary inclusion of Origanum vulgare extract on non-specific immune responses and hematological parameters of rainbow trout (Oncorhynchus mykiss). BEPLS 4:33–39Google Scholar
  62. Rajaian H, Jalaee J, Aghajani A (2006) Berberis vulgaris as growth promoter in broiler chickens. Int J Poult Sci 5:395–397CrossRefGoogle Scholar
  63. Reverter M, Bontemps N, Lecchini D, Banaigs B, Sasal P (2014) Use of plant extracts in fish aquaculture as an alternative to chemotherapy: current status and future perspectives. Aquaculture 433:50–61CrossRefGoogle Scholar
  64. Romero J, Feijoó CG, Navarrete P (2012) Antibiotics in aquaculture–use, abuse and alternatives. In: Health and environment in aquaculture. InTechGoogle Scholar
  65. Roohani AM, Abedian Kenari A, Fallah Kapoorchali M, Borani MS, Zoriezahra SJ, Smailey AH, Esmaeili M, Rombenso AN (2019) Effect of spirulina Spirulina platensis as a complementary ingredient to reduce dietary fish meal on the growth performance, whole-body composition, fatty acid and amino acid profiles, and pigmentation of Caspian brown trout (Salmo trutta caspius) juveniles. Aquac Nutr 25:633–645CrossRefGoogle Scholar
  66. Santos L, Ramos F (2018) Antimicrobial resistance in aquaculture: current knowledge and alternatives to tackle the problem. Int J Antimicrob Agents 52:135–143CrossRefGoogle Scholar
  67. Sarraf M, Beig-babaei A, Naji-Tabasi S (2019) Investigating functional properties of barberry species: an overview. J Sci Food Agric (in press)Google Scholar
  68. Sepehrimanesh M, Poorbaghi SL, Rajaian H, Dadras H, Razeghian Jahromi I (2012) Effects of addition of Berberis vulgaris root powder to the arbor acers chicks ration as a growth promoter. J Med Plants 2:130–136Google Scholar
  69. Stermitz FR, Beeson TD, Mueller PJ, Hsiang J-F, Lewis K (2001) Staphylococcus aureus MDR efflux pump inhibitors from a Berberis and a Mahonia (Sensu strictu) species. Biochem Syst Ecol 29:793–798CrossRefGoogle Scholar
  70. Stratev D, Zhelyazkov G, Noundou XS, Krause RW (2018) Beneficial effects of medicinal plants in fish diseases. Aquac Int 26:289–308CrossRefGoogle Scholar
  71. Taheri S, Zarei A, Ashtiyani SC, Rezaei A, Zaheiri S (2012) Evaluation of the effects of hydroalcoholic extract of Berberis vulgaris root on the activity of liver enzymes in male hypercholesterolemic rats. Avicenna J Phytomed 2:153–161Google Scholar
  72. Tomosaka H, Chin YW, Salim AA, Keller WJ, Chai H, Kinghorn AD (2008) Antioxidant and cytoprotective compounds from Berberis vulgaris (barberry). Phytother Res 22:979–981CrossRefGoogle Scholar
  73. Tukmechi A, Morshedi A, Delirezh N (2007) Changes in intestinal microflora and humoral immune response following probiotic administration in rainbow trout (Oncorhynchus mykiss). J Anim Vet Adv 6:1183–1189Google Scholar
  74. Tung Y-T, Wu J-H, Hsieh C-Y, Chen P-S, Chang S-T (2009) Free radical-scavenging phytochemicals of hot water extracts of Acacia confusa leaves detected by an on-line screening method. Food Chem 115:1019–1024CrossRefGoogle Scholar
  75. Van Hai N (2015) The use of medicinal plants as immunostimulants in aquaculture: a review. Aquaculture 446:88–96CrossRefGoogle Scholar
  76. Vaseeharan B, Thaya R (2014) Medicinal plant derivatives as immunostimulants: an alternative to chemotherapeutics and antibiotics in aquaculture. Aquac Int 22:1079–1091CrossRefGoogle Scholar
  77. Vazirzadeh A, Dehghan F, Kazemeini R (2017) Changes in growth, blood immune parameters and expression of immune related genes in rainbow trout (Oncorhynchus mykiss) in response to diet supplemented with Ducrosia anethifolia essential oil. Fish Shellfish Immunol 69:164–172CrossRefGoogle Scholar
  78. Villinski J, Dumas E, Chai H-B, Pezzuto J, Angerhofer C, Gafner S (2003) Antibacterial activity and alkaloid content of Berberis thunbergii, Berberis vulgaris and Hydrastis canadensis. Pharm Biol 41:551–557CrossRefGoogle Scholar
  79. Wenk C (2003) Herbs and botanicals as feed additives in monogastric animals. Asian Australas J Anim Sci 16:282–289CrossRefGoogle Scholar
  80. Xiong W, Sun Y, Zhang T, Ding X, Li Y, Wang M, Zeng Z (2015) Antibiotics, antibiotic resistance genes, and bacterial community composition in fresh water aquaculture environment in China. Microb Ecol 70:425–432CrossRefGoogle Scholar
  81. Yazdani A, Poorbaghi SL, Habibi H, Nazifi S, Far FR, Sepehrimanesh M (2013) Dietary Berberis vulgaris extract enhances intestinal mucosa morphology in the broiler chicken (Gallus gallus). Comp Clin Pathol 22:611–615CrossRefGoogle Scholar
  82. Zeilab Sendijani R, Abedian Kenari A, Smiley AH, Esmaeili M (2019) The effect of extract from dill (Anethum graveolens) on the growth performance, body composition, immune system and antioxidant system of rainbow trout (Oncorhynchus mykiss). N Am J Aquac (in press)Google Scholar
  83. Zhou W, Rahimnejad S, Lu K, Wang L, Liu W (2018) Effects of berberine on growth, liver histology, and expression of lipid-related genes in blunt snout bream (Megalobrama amblycephala) fed high-fat diets. Fish Physiol Biochem 1–9Google Scholar

Copyright information

© Springer-Verlag London Ltd., part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Aquaculture, Faculty of Natural Resources and Marine SciencesTarbiat Modares UniversityNoorIran
  2. 2.Institute for Marine and Antarctic StudiesUniversity of TasmaniaHobartAustralia

Personalised recommendations