Effect of garlic extract supplementation on growth performance, nonspecific immunity, and antibacterial activity of skin mucus in goldfish, Carassius auratus

Hamidreza Ahmadniaye Motlagh, omid Safari, Sajjad Pourmozaffar


In the present study, the effects of dietary supplementation of garlic extract on growth performance, skin mucus immunological parameters and antibacterial activity of Carassius auratus were examined. Fish were stocked in 100 L glass tanks (6 fish per tank) in triplicate and fed diets containing different garlic extracts (0 (control), 5, 10, and 15 ml/kg] for eight weeks. At the end of feeding period, the fish skin mucus was collected for evaluating the components of non-specific immune system (including lysozyme, complement, total immunoglobulin, dissolved protein, and alkaline phosphatase). Additionally, antimicrobial activity of the skin mucus against Aeromonas hydrophila, Yersinia ruckeri, Micrococcus luteus, Streptococcus faecium, and S. iniae was assessed. After the feeding trial, the fish fed diets containing garlic extract showed no significant difference in growth parameters. Significantly higher skin mucus lysozyme, complement, alkaline phosphatase activities, and total immunoglobulin and dissolved protein concentration were observed in the fish fed garlic extract-supplemented diets (P<0.0001). The antimicrobial activity of the skin mucus increased along with the increase in the dietary garlic extract levels (P<0.0001). Moreover, garlic extract exhibited the antimicrobial activity against pathogenic bacterial species. The highest level of dietary garlic extract (15 ml/kg) led to significantly higher inhibition zones against pathogenic bacterial species compared to the other garlic extract levels (P<0.0001). The optimal administration of garlic extract at 15 ml/kg enhance skin mucus immune parameters and antimicrobial activity in goldfish.


Antimicrobial activity, Garlic extract, Growth Immune parameters, Skin mucus.

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Adineh H., Harsij M., Jafaryan H., Asadi M. (2020). The effects of microencapsulated garlic (Allium sativum) extract on growth performance, body composition, immune response and antioxidant status of rainbow trout (Oncorhynchus mykiss) juveniles. Journal of Applied Animal Research, 48(1): 372-378.

Ahmadniaye Motlagh H., Safari O., Paolucci M. (2019). Effect of different levels of milkweed (Calotropis persica) seed powder on the growth parameters, immunity and gut microbiota of Oncorhynchus mykiss. Iranian Journal of Veterinary Science and Technology, 11(1): 43-50.

Ahmadniaye Motlagh H., Safari O., Selahvarzi Y., Baghalian A., Kia E. (2020). Non-specific immunity promotion in response to garlic extract supplemented diets in female Guppy (Poecilia reticulata). Fish and Shellfish Immunology, 97(July): 96-99.

Aly S.M., Mohamed M.F. (2010). Echinacea purpurea and Allium sativum as immunostimulants in fish culture using Nile tilapia (Oreochromis niloticus). Journal of Animal Physiology and Animal Nutrition, 94(5): 31-39.

Arreola R., Quintero-Fabián S., López-Roa R., Flores-Gutiérrez E., Reyes-Grajeda J., Carrera-Quintanar L., Ortuño-Sahagún D. (2015). Immunomodulation and anti-inflammatory effects of garlic compounds. Journal of Immunology Research, 2015: 1-13.

Bhatwalkar S.B., Mondal R., Krishna S.B.N., Adam J.K., Govender P., Anupam R. (2021). antibacterial properties of organosulfur compounds of garlic (Allium sativum). Frontiers in Microbiology, 12: 1-20.

Breyer K.E., Getchell R.G., Cornwell E.R., Wooster G.A., Ketola H.G., Bowser P.R. (2015). Efficacy of an extract from garlic, Allium sativum, against infection with the Furunculosis bacterium, Aeromonas salmonicida, in rainbow trout, Oncorhynchus mykiss. Journal of the World Aquaculture Society, 46(3): 273-282.

Castillo S., Rosales M., Pohlenz C., Gatlin D.M. (2014). Effects of organic acids on growth performance and digestive enzyme activities of juvenile red drum Sciaenops ocellatus. Aquaculture, 433: 6-12.

Chen Y.Y., Chen J.C., Kuo Y.H., Lin Y.C., Chang Y.H., Gong H.Y., Huang C.L. (2016). Lipopolysaccharide and β-1,3-glucan-binding protein (LGBP) bind to seaweed polysaccharides and activate the prophenoloxidase system in white shrimp Litopenaeus vannamei. Developmental and Comparative Immunology, 55: 144-151.

Daniels C.L., Merrifield D.L., Ringø E., Davies S.J. (2013). Probiotic, prebiotic and synbiotic applications for the improvement of larval European lobster (Homarus gammarus) culture. Aquaculture, 416-417: 396-406.

Fierro-Coronado J.A., Angulo C., Rubio-Castro A., Luna-González A., Cáceres-Martínez C.J., Ruiz-Verdugo C.A., Diarte-Plata G. (2018). Dietary fulvic acid effects on survival and expression of immune-related genes in Litopenaeus vannamei challenged with Vibrio parahaemolyticus. Aquaculture Research, 49(9): 3218-3227.

Ghehdarijani M.S., Hajimoradloo A., Ghorbani R., Roohi Z. (2016). The effects of garlic-supplemented diets on skin mucosal immune responses, stress resistance and growth performance of the Caspian roach (Rutilus rutilus) fry. Fish and Shellfish Immunology, 49: 79-83.

Gholipour Kanani H., Nobahar Z., Kakoolaki S., Jafarian H. (2014). Effect of ginger and garlic-supplemented diet on growth performance, some hematological parameters and immune responses in juvenile Huso huso. Fish Physiology and Biochemistry, 40(2): 481-490.

Guo J.J., Kuo C.M., Chuang Y.C., Hong J.W., Chou R.L., Chen T.I. (2012). The effects of garlic-supplemented diets on antibacterial activity against Streptococcus iniae and on growth in orange-spotted grouper, Epinephelus coioides. Aquaculture, 364-365: 33-38.

Harikrishnan R., Balasundaram C., Kim M.C., Kim J.S., Han Y.J., Heo M.S. (2009). Innate immune response and disease resistance in Carassius auratus by triherbal solvent extracts. Fish and Shellfish Immunology, 27(3): 508-515.

Harikrishnan R., Kim M.C., Kim J.S., Balasundaram C., Heo M.S. (2011). Protective effect of herbal and probiotics enriched diet on haematological and immunity status of Oplegnathus fasciatus (Temminck & Schlegel) against Edwardsiella tarda. Fish and Shellfish Immunology, 30(3): 886-893.

Hoseini S.M., Gharavi B., Mirghaed A.T., Hoseinifar S.H., Van Doan H. (2021). Effects of dietary phytol supplementation on growth performance, immunological parameters, antioxidant and stress responses to ammonia exposure in common carp, Cyprinus carpio (Linnaeus, 1758). Aquaculture, 545: 737151.

Hoseini S.M., Mirghaed A.T., Iri Y., Ghelichpour M. (2018). Effects of dietary cineole administration on growth performance, hematological and biochemical parameters of rainbow trout (Oncorhynchus mykiss). Aquaculture, 495: 766-772.

Hoseini S.M., Yousefi M. (2019). Beneficial effects of thyme (Thymus vulgaris) extract on oxytetracycline-induced stress response, immunosuppression, oxidative stress and enzymatic changes in rainbow trout (Oncorhynchus mykiss). Aquaculture Nutrition, 25(2): 298-309.

Hoseinifar S.H., Khalili M., Rufchaei R., Raeisi M., Attar M., Cordero H., Esteban M.Á. (2015). Effects of date palm fruit extracts on skin mucosal immunity, immune related genes expression and growth performance of common carp (Cyprinus carpio) fry. Fish and Shellfish Immunology, 47(2): 706-711.

Hoseinifar S.H., Shakouri M., Doan H., Van, Shafiei S., Yousefi M., Raeisi M., Raeisi M., Yousefi S., Harikrishnan R., Reverter M. (2020). Dietary supplementation of lemon verbena (Aloysia citrodora) improved immunity, immune-related genes expression and antioxidant enzymes in rainbow trout (Oncorrhyncus mykiss). Fish and Shellfish Immunology, 99: 379-385.

Hoseinifar S.H., Zoheiri F., Dadar M., Rufchaei R., Ringø E. (2016). Dietary galactooligosaccharide elicits positive effects on non-specific immune parameters and growth performance in Caspian white fish (Rutilus frisii kutum) fry. Fish and Shellfish Immunology, 56: 467-472.

Tamadoni Jahromi S., Pourmozaffar S., Rameshi H., Gozari M., Nahavandi R. (2020). Evaluation of hemolymph biochemical properties, clearance rate, bacterial microbiota and expression of HSP genes of gulf pearl oyster Pinctada radiata in response to salinity changes. Fisheries Science, 86(6): 1055-1065.

Tamadoni Jahromi S., Pourmozaffar S., Jahanbakhshi A., Rameshi H., Gozari M., Khodadadi M., Sohrabipour J., Behzadi S., Barzkar N., Nahavandi R., Zahedi M.R., Moezzi M. (2021). Effect of different levels of dietary Sargassum cristaefolium on growth performance, hematological parameters, histological structure of hepatopancreas and intestinal microbiota of Litopenaeus vannamei. Aquaculture, 533: 1-17.

Karimi Pashaki A., Zorriehzahra S.M.J., Ghasemi M., Sharif Rohani M., Hosseini S. (2018). Effects of dietary garlic extract on some blood, immunity and growth parameters of common carp fingerlings (Cyprinus carpio). Iranian Journal of Aquatic Animal Health, 4(2): 28-39.

Labrador J.R.P., Guinares R.C., Hontiveros G.J.S. (2016). Effect of garlic powder-supplemented diets on the growth and survival of Pacific white leg shrimp (Litopenaeus vannamei). Cogent Food and Agriculture, 2(1): 1-8.

Lallès J.P. (2019). Biology, environmental and nutritional modulation of skin mucus alkaline phosphatase in fish: A review. Fish and Shellfish Immunology, 89(January): 179-186.

Lee D.H., Lim S.R., Han J.J., Lee S.W., Ra C.S., Kim J.D. (2014). Effects of dietary garlic powder on growth, feed utilization and whole body composition changes in fingerling sterlet sturgeon, Acipenser ruthenus. Asian-Australasian Journal of Animal Sciences, 27(9): 1303-1310.

Li F., An X., Wu D., Xu J., Chen Y., Li W., Cao Y., Guo X., Lin X., Li C., Liu S., Song H. (2019). Engineering microbial consortia for high-performance cellulosic hydrolyzates-fed microbial fuel cells. Frontiers in Microbiology, 10(Mar): 1-10.

Lowry O., Rosebrough N., Farr A.L., Randall R. (1951). Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry, 193(1): 265-275.

Magarinos B., Pazos F., Santos Y., Romalde J.L., Toranzo A.E. (1995). Response of Pasteurella piscicida and Flexibacter maritimus to skin mucus of marine fish. Diseases of Aquatic Organisms, 21(2): 103-108.

Mahmoud R., Aziza A., Marghani B., Eltaysh R. (2019). Influence of ginger and garlic supplementation on growth performance, whole body composition and oxidative stress in the muscles of Nile tilapia (Oreochromis niloticus). Advances in Animal and Veterinary Sciences, 7(5): 397-404.

Mansouri Taee H., Hajimoradloo A., Hoseinifar S.H., Ahmadvand H. (2017). Dietary Myrtle (Myrtus communis L.) improved non-specific immune parameters and bactericidal activity of skin mucus in rainbow trout (Oncorhynchus mykiss) fingerlings. Fish and Shellfish Immunology, 64 (March): 320-324.

Mimeault C., Woodhouse A.J., Miao X.S., Metcalfe C.D., Moon T.W., Trudeau V.L. (2005). The human lipid regulator, gemfibrozil bioconcentrates and reduces testosterone in the goldfish, Carassius auratus. Aquatic Toxicology, 73(1): 44-54.

Natasya-Ain R., Eirna-Liza N., Jasmin M.Y., Karim M. (2018). Antibacterial activity of garlic extracts on fish pathogenic bacteria. Journal of Environmental Biology, 39(5): 808-812.

Ndong D., Fall J. (2011). The effect of garlic (Allium sativum) on growth and immune responses of hybrid tilapia (Oreochromis niloticus x Oreochromis aureus). Journal of Clinical Immunology and Immunopathology Research, 3(1): 1-9.

Nya E.J., Austin B. (2009). Use of garlic, Allium sativum, to control Aeromonas hydrophila infection in rainbow trout, Oncorhynchus mykiss (Walbaum). Journal of Fish Diseases, 32(11): 963-970.

Nya E.J., Dawood Z., Austin B. (2010). The garlic component, allicin, prevents disease caused by Aeromonas hydrophila in rainbow trout, Oncorhynchus mykiss (Walbaum). Journal of Fish Diseases, 33(4): 293-300.

Paray B.A., Hoseini S.M., Hoseinifar S.H., Van Doan H. (2020). Effects of dietary oak (Quercus castaneifolia) leaf extract on growth, antioxidant, and immune characteristics and responses to crowding stress in common carp (Cyprinus carpio). Aquaculture, 524: 735276.

Pourmozaffar S., Hajimoradloo A., Paknejad H., Rameshi H. (2019). Effect of dietary supplementation with apple cider vinegar and propionic acid on hemolymph chemistry, intestinal microbiota and histological structure of hepatopancreas in white shrimp, Litopenaeus vannamei. Fish and Shellfish Immunology, 86: 900-905.

Sahu S., Das B.K., Mishra B.K., Pradhan J., Sarangi N. (2007). Effect of Allium sativum on the immunity and survival of Labeo rohita infected with Aeromonas hydrophila. Journal of Applied Ichthyology, 23(1): 80-86.

Sarhadi I., Alizadeh E., Ahmadifar E., Adineh H., Dawood M.A.O. (2020). Skin mucosal, serum immunity and antioxidant capacity of common carp (Cyprinus carpio) fed Artemisia (Artemisia annua). Annals of Animal Science, 20 (3): 1011-1027.

Sharma V., Sharma A., Kansal L. (2010). The effect of oral administration of Allium sativum extracts on lead nitrate induced toxicity in male mice. Food and Chemical Toxicology, 48(3): 928-936.

Sheikhzadeh N., Heidarieh M., Karimi Pashaki A., Nofouzi K., Ahrab Farshbafi M., Akbari M. (2012). Hilyses®, fermented Saccharomyces cerevisiae, enhances the growth performance and skin non-specific immune parameters in rainbow trout (Oncorhynchus mykiss). Fish and Shellfish Immunology, 32(6): 1083-1087.

Stolen J.S., Fletcher T.C., Rowley A.F., Zelikoff J.T., Kaattari S.L., Smith S.A. (1994). Techniques in fish immunology. SOS publications. 222 p.

Sukumaran V., Park S.C., Giri S.S. (2016). Role of dietary ginger Zingiber officinale in improving growth performances and immune functions of Labeo rohita fingerlings. Fish and Shellfish Immunology, 57: 362-370.

Talpur A.D., Ikhwanuddin M. (2012). Dietary effects of garlic (Allium sativum) on haemato-immunological parameters, survival, growth, and disease resistance against Vibrio harveyi infection in Asian sea bass, Lates calcarifer (Bloch). Aquaculture, 364-365: 6-12.

Tanekhy M., Fall J. (2015). Expression of innate immunity genes in kuruma shrimp Marsupenaeus japonicus after in vivo stimulation with garlic extract (allicin). Veterinarni Medicina, 60(1): 39-47.

Ushimaru P.I., Da Silva M.T.N., Di Stasi L.C., Barbosa L., Fernandes A. (2007). Antibacterial activity of medicinal plant extracts. Brazilian Journal of Microbiology, 38(4): 717-719.

Vaseeharan B., Prasad G.S., Ramasamy P., Brennan G. (2011). Antibacterial activity of Allium sativum against multidrug-resistant Vibrio harveyi isolated from black gill-diseased Fenneropenaeus indicus. Aquaculture International, 19(3): 531-539.

Vuddhakul V., Bhoopong P., Hayeebilan F., Subhadhirasakul S. (2007). Inhibitory activity of Thai condiments on pandemic strain of Vibrio parahaemolyticus. Food Microbiology, 24(4): 413-418.

Wang C., Lu S., Li J., Wang L., Jiang H., Liu Y., Yin J. (2020). Effects of dietary myo-inositol on growth, antioxidative capacity, and nonspecific immunity in skin mucus of taimen Hucho taimen fry. Fish Physiology and Biochemistry, 46(3): 1011-1018.

Yin X.L., Li Z.J., Yang K., Lin H.Z., Guo Z.X. (2014). Effect of guava leaves on growth and the non-specific immune response of Penaeus monodon. Fish and Shellfish Immunology, 40(1): 190-196.

Yogeeswaran A., Velmurugan S., Punitha S.M.J., Babu M.M., Selvaraj T., Kumaran T., Citarasu T. (2012). Protection of Penaeus monodon against white spot syndrome virus by inactivated vaccine with herbal immunostimulants. Fish and Shellfish Immunology, 32(6): 1058-1067.

Yousefi M., Shabunin S.V., Vatnikov Y.A., Kulikov E.V., Adineh H., Hamidi M.K., Hoseini S.M. (2020a). Effects of lavender (Lavandula angustifolia) extract inclusion in diet on growth performance, innate immunity, immune-related gene expression, and stress response of common carp, Cyprinus carpio. Aquaculture, 515: 734588.

Yousefi M., Vatnikov Y.A., Kulikov E.V., Plushikov V.G., Drukovsky S.G., Hoseinifar S.H., Van Doan H. (2020b). The protective effects of dietary garlic on common carp (Cyprinus carpio) exposed to ambient ammonia toxicity. Aquaculture, 526: 735400.

Yu M.C., Li Z.J., Lin H.Z., Wen G.L., Ma S. (2009). Effects of dietary medicinal herbs and Bacillus on survival, growth, body composition, and digestive enzyme activity of the white shrimp Litopenaeus vannamei. Aquaculture International, 17(4): 377-384.

Zaefarian A., Yeganeh S., Adhami B. (2017). Dietary effects of garlic powder (Allium sativum) on growth, blood indices, carcass composition, and lysozyme activity in brown trout (Salmo caspius) and resistance against Yersinia ruckeri infection. Aquaculture International, 25(6): 1987-1996.

Zargar A., Taheri Mirghaed A., Mirzargar S.S., Ghelichpour M., Yousefi M., Hoseini S.M. (2020). Dietary ginger administration attenuates oxidative stress and immunosuppression caused by oxytetracycline in rainbow trout (Oncorhynchus mykiss). Aquaculture Research, 51(10): 4215-4224.

Zheng Z.L., Tan J.Y.W., Liu H.Y., Zhou X.H., Xiang X., Wang K.Y. (2009). Evaluation of oregano essential oil (Origanum heracleoticum L.) on growth, antioxidant effect and resistance against Aeromonas hydrophila in channel catfish (Ictalurus punctatus). Aquaculture, 292(3-4): 214-218.


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