This study aimed to evaluate the in vitro effect of carvacrol on different microorganisms of importance in shrimp farming, as well as its in vivo effect on zootechnical, immunological and microbiological performance, as well as resistance, of Litopenaeus vannamei challenged with Vibrio parahaemolyticus. In particular, the antimicrobial activity of carvacrol was evaluated in vitro by analysis of the minimum inhibitory concentration (MIC) and by agar diffusion disc with Gram-negative and Gram-positive bacteria. The in vivo experiment was conducted using different concentrations of carvacrol (1, 3, 4 and 6 mg mL-1) added to shrimp feed, together with a control diet without carvacrol. After four weeks, zootechnical, immunological and microbiological parameters, as well as resistance, of animals challenged with V. parahaemolyticus were evaluated. The MIC of Vibrio alginolyticus and Vibrio harveyi was 0.078 mg mL-1, while for the other bacteria, it was 0.156 mg mL-1 of carvacrol. The greatest halos of inhibition were observed in V. parahaemolyticus and Vibrio harveyi with significant differences demonstrated for the other microorganisms, except Escherichia coli. The in vivo results showed no significant differences among treatments. In conclusion, the antimicrobial activity of carvacrol was confirmed with Gram-negative and Gram-positive bacteria, and it is suggested that its antimicrobial potential is more effective against Vibrio spp. However, the concentrations of carvacrol used in vivo did not affect the parameters evaluated.


ABCC - Associação Brasileira de Criadores de Camarão. 2017. Vamos reforçar a defesa sanitária do Brasil ou vamos permitir a importação de camarão do Equador?, (1): 18. Disponivel em . Acessado em 16 set., 2020.

AOAC - Association of Official Agricultural Chemists. 2005. Official methods of analysis of AOAC International. 18th ed. AOAC International Ed. Maryland, USA, 1094 p.

Abutbul, S.; Golan-Goldhirsh, A.; Zilberg, D. 2004. Use of Rosmarinus officinalis as a treatment against Streptococcus iniae in tilapia (Oreochromis sp.). Aquaculture, 238(1–4): 97–105.

Ahmadifar, E.; Falahatkar, B.; Akrami, R. 2011. Effects of dietary thymol-carvacrol on growth performance, hematological parameters and tissue composition of juvenile rainbow trout, Oncorhynchus mykiss. Journal of Applied Ichthyology, 27(4): 1057–1060.

Araujo, M.M.; Longo, P.L. 2016. Teste da ação antibacteriana in vitro de óleo essencial comercial de Origanum vulgare (orégano) diante das cepas de Escherichia coli e Staphylococcus aureus. Arquivos do Instituto Biológico, 83: 1–7.

Astashkina, A.; Mann, B.; Grainger, D.W. 2012. A critical evaluation of in vitro cell culture models for high-throughput drug screening and toxicity. Pharmacology and Therapeutics, 134 (1): 82–106.

Baruah, K.; Norouzitallab, P.; Phong, H.P.PD.; Smagghe, G.; Bossier, P. 2017. Enhanced resistance against Vibrio harveyi infection by carvacrol and its association with the induction of heat shock protein 72 in gnotobiotic Artemia franciscana. Cell Stress and Chaperones, 22(3): 377–387.

Bimczok, D.; Rau, H.; Sewekow, E.; Janczyk, P.; Souffrant, W.B.; Rothkötter, H.J. 2008. Influence of carvacrol on proliferation and survival of porcine lymphocytes and intestinal epithelial cells in vitro. Toxicology in Vitro, 22(3): 652–658.

Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72 (1–2): 248–254.

Carvalho, R.I.; Medeiros, A.S.J.; Chaves, M.; Souza, E.L.; Magnani, M. 2018. Lipids, pH, and their interaction affect the inhibitory effects of carvacrol against Salmonella Typhimurium PT4 and Escherichia coli O157:H7. Frontiers in Microbiology, 8: 2701.

Chakraborty, S.B.; Hancz, C. 2011. Application of phytochemicals as immunostimulant, antipathogenic and antistress agents in finfish culture. Reviews in Aquaculture, 3(3): 103–119.

Chavan, P.S.; Tupe, S.G. 2014. Antifungal activity and mechanism of action of carvacrol and thymol against vineyard and wine spoilage yeasts. Food Control, 46: 115–120.

Cosentino, S.; Tuberoso, C.I.G.; Pisano, B.; Satta, M.; Mascia, V.; Arzedi, E.; Palmas, F. 1999. In-vitro antimicrobial activity and chemical composition of Sardinian Thymus essential oils. Letters in Applied Microbiology, 29(2): 130–135.

Fang, S.; Zhou, Q.; Hu, Y.; Liu, F.; Mei, J.; Xie, J. 2019. Antimicrobial carvacrol incorporated in flaxseed gum-sodium alginate active films to improve the quality attributes of Chinese sea bass (Lateolabrax maculatus) during cold storage. Molecules, 24(18): 1–17.

FAO - Food and Agriculture Organization of the United Nations, 2020. The state of world fisheries and aquaculture 2020: sustainability in action. Rome: SOFIA. Available from: Accessed: 10 Nov., 2020.

FDA - Food and Drug Administration. 2017. CRF - Code of federal regulations title 21, 3: 25 - 26.

Garrett, W.S.; Gordon, J.I.; Glimcher, L.H. 2010. Homeostasis and Inflammation in the Intestine. Cell, 140 (6): 859–870.

Gilles, M.; Zhao, J.; Na, M.; Agboola, S. 2010. Chemical composition and antimicrobial properties of essential oils of three Australian Eucalyptus species. Food Chemistry, 119(2): 731–737.

Gong, H.; Lawrence, A.L.; Jiang, D.H.; Castille, F.L.; Gatlin, D.M. 2000. Lipid nutrition of juvenile Litopenaeus vannamei: I. Dietary cholesterol and de-oiled soy lecithin requirements and their interaction. Aquaculture, 190(3–4): 305–324.

Gracia-Valenzuela, M.H.; Vergara-Jiménez, M.J.; Baez-Flores, M.E.; Cabrera-Chavez, F. 2014. Antimicrobial effect of dietary oregano essential oil against vibrio bacteria in shrimps. Archives of Biological Sciences, 66(4): 1367–1370.

Guarda, A.; Rubilar, J.F.; Miltz, J.; Galotto, M.J. 2011. The antimicrobial activity of microencapsulated thymol and carvacrol. International Journal of Food Microbiology, 146(2): 144–150.

Guimarães, A.G.; Oliveira, G.F.; Melo, M.S.; Cavalcanti, S.C.H.; Antoniolli, A.R.; Bonjardim, L.R.; Silva, F.A.; Santos, J.P.; Rocha, R.F.; Moreira, J.C. F.; Araujo, A.A.S.; Gelain, D.P.; Quintans-junior, L.J. 2010. Bioassay-guided evaluation of antioxidant and antinociceptive activities of carvacrol. Basic and Clinical Pharmacology and Toxicology, 107(6): 949–957.

Hajlaoui, H.; Snoussi, M.; Noumi, E.; Zanetti, S.; Ksouri, R.; Bakhrouf A. 2010. Chemical composition, antioxidant and antibacterial activities of the essential oils of five Tunisian aromatic plants. Italian Journal of Food Science, 22(3): 320–329.

Inamuco, J.; Veenendaal, A.K.J.; Burt, S.A.; Post, J.A.; Bokhoven, J.L.M.T.V.; Haagsman, H.P.; Veldhuizen, E.J.A. 2012. Sub-lethal levels of carvacrol reduce Salmonella Typhimurium motility and invasion of porcine epithelial cells. Veterinary Microbiology, 157(1–2): 200–207.

Knobloch, K.; Pauli, A.; Iberl, B.; Weigand, H.; Weis, N. 1989. Antibacterial and Antifungal Properties of Essential Oil Components. Journal of Essential Oil Research, 1(3): 119-128.

Lemos, M.F.; Lemos, M.F.; Pacheco, H.P.; Guimarães, A.C.; Fronza, M.; Endringer, D.C.; Scherer, R. 2017. Seasonal variation affects the composition and antibacterial and antioxidant activities of Thymus vulgaris. Industrial Crops and Products, 95: 543–548.

Lima, D.S.; Lima, J.C.; Calvacanti, R.M.C.B.; Santos, B.H.C.; Lima, I.O. 2017. Estudo da atividade antibacteriana dos monoterpenos timol e carvacrol contra cepas de Escherichia coli produtoras de β-lactamases de amplo espectro. Revista Pan-Amazônica de Saúde, 8(1): 17–21.

Maggioni, D.S; Andreatta, E.R.; Hermes, E.M.; Barracco, M.A. 2004. Evaluation of some hemato-immunological parameters in female shrimp Litopenaeus vannamei submitted to unilateral eyestalk ablation in association with a diet supplemented with superdoses of ascorbic acid as a form of immunostimulation. Aquaculture, 241(1–4): 501–515.

Marinelli, L.; Stefano, A.D.; Cacciatore, I. 2018. Carvacrol and its derivatives as antibacterial agents. Phytochemistry Reviews, 17(4): 903–921.

NCCLS - National Committee for Clinical Laboratory Standards. 2006. Methods for Broth Dilution Susceptibility Testing of Bacteria Isolated from Aquatic Animals; Approved Guideline. NCCLS document M49-A, 26. Wayne, Pennsylvania, U.S.A.

Nevas, M.; Korhonen, A.R.; Lindström, M.; Turkki, P.; Korkeala, H. 2004. Antibacterial efficiency of Finnish spice essential oils against pathogenic and spoilage bacteria. Journal of food protection, 67(1): 199–202.

Nostro, A.; Blanco, A.R.; Cannatelli, M.A.; Enea, V.; Flamini, G.; Morelli, I.; Roccaro, A.S.; Alonzo, V. 2004. Susceptibility of methicillin-resistant staphylococci to oregano essential oil, carvacrol and thymol. FEMS Microbiology Letters, 230(2): 191–195.

Nostro, A.; Cellini, L.; Zimbalatti, V.; Blanco A.R.; Marino, A.; Pizzimenti, F.; Giulio, M.D.; Bisignano, G. 2012. Enhanced activity of carvacrol against biofilm of Staphylococcus aureus and Staphylococcus epidermidis in an acidic environment. Acta pathologica, microbiologica et immunologica scandinavica, 120(12): 967–973.

NRC - National Research Council. 2011. Nutrient Requirements of Fish and Shrimp. Washington: National Academies Press, 390p.

Rattanachaikunsopon, P.; Phumkhachorn, P. 2009. Prophylactic effect of Andrographis paniculata extracts against Streptococcus agalactiae infection in Nile tilapia (Oreochromis niloticus). Journal of Bioscience and Bioengineering, 107(5): 579–582.

Rattanachaikunsopon, P.; Phumkhachorn, P. 2010. Assessment of synergistic efficacy of carvacrol and cymene against Edwardsiella tarda in vitro and in Tilapia ( Oreochromis niloticus). African Journal of Microbiology Research, 4(5): 420–425.

Ray, A.J.; Lewis, B.L.; Browdy, C.L.; Leffler, J.W. 2010. Suspended solids removal to improve shrimp (Litopenaeus vannamei) production and an evaluation of a plant-based feed in minimal-exchange, superintensive culture systems. Aquaculture, 299(1–4): 89–98.

Rombout J.H.W.M.; Abelli, L.; Picchietti, S.; Scapigliati, G.; Kiron, V. 2011. Teleost intestinal immunology. Fish and Shellfish Immunology, 31(5): 616–626.

Roselli, M.; Britti, M.S.; Huërou-Luron, I.L.; Marfaing, H.; Zhu, W.Y.; Mengheri, E. 2007. Effect of different plant extracts and natural substances (PENS) against membrane damage induced by enterotoxigenic Escherichia coli K88 in pig intestinal cells. Toxicology in Vitro, 21(2): 224–229.

Samy, R.P.; Gopalakrishnakone, P. 2010. Therapeutic potential of plants as anti-microbials for drug discovery. Evidence-based Complementary and Alternative Medicine, 7(3): 283–294.

Sánchez, C.; Aznar, R.; Sánchez, G. 2015. The effect of carvacrol on enteric viruses. International Journal of Food Microbiology, 192: 72–76

Santos, H.M.; Tsai, C.; Maquiling, K.R.A.; Tayo, L.L.; Mariatulqabtiah, A.R.; Lee, C.; Chuang, K.P. 2020. Diagnosis and potential treatments for acute hepatopancreatic necrosis disease (AHPND): a review. Aquaculture International, 28: 169-185.

Sikkema, J.; Bont, J.A.; Poolman, B. 1995. Mechanisms of membrane toxicity of hydrocarbons. Microbiological review, 59: 201–222.

Snoussi, M.; Hajlaoui, H.; Noumi, E.; Usai, D.; Sechi, L.A.; Zanetti, S.; Bakhrouf, A. 2008. In-vitro anti-Vibrio spp. activity and chemical composition of some Tunisian aromatic plants. World Journal of Microbiology and Biotechnology, 24(12): 3071–3076.

Söderhäll, K.; Häll, L. 1984. Lipopolysaccharide-induced activation of prophenoloxidase activating system in crayfish haemocyte lysate. Biochimica et Biophysica Acta (BBA) - General Subjects, 797(1): 99–104.

Stratakos, A.C.; Sima, F.; Ward, P.; Linton, M.; Kelly, C.; Pinkerton, L.; Stef, L.; Pet, I.;
Corcionivoschi, N. 2018. The in vitro effect of carvacrol, a food additive, on the pathogenicity of O157 and non-O157 Shiga-toxin producing Escherichia coli. Food Control, 84: 290–296.

Talpur, A.D.; Ikhwanuddin, M.; Bolong, A.M.A. 2013. Nutritional effects of ginger (Zingiber officinale Roscoe) on immune response of Asian sea bass, Lates calcarifer (Bloch) and disease resistance against Vibrio harveyi. Aquaculture, 400–401: 46–52.

Van wyk, P.; Scarpa, J. 1999. Water Quality Requirements and Management. In: Van Wyk, P. et al. (Eds.). Farming Marine Shrimp in Recirculating Freshwater Systems. Florida: Department of Agriculture and Consumer Services. 141–162p.

Volpatti, D.; Bulfon, C.; Tulli, F.; Galeotti, M. 2013. Growth parameters, innate immune response and resistance to Listonella (Vibrio) anguillarum of Dicentrarchus labrax fed carvacrol supplemented diets. Aquaculture Research, 45(1): 31–44.

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.

Zhou, Q.C.; Zeng, W.P.; Wang, H.L.; Wang, T.; Wang, Y.L.; Xie, F.J. 2012. Dietary arginine requirement of juvenile Pacific white shrimp, Litopenaeus vannamei. Aquaculture, 364–365: 252–258.
How to Cite
COELHO, Jaqueline da Rosa et al. IN VITRO ANTIMICROBIAL ACTIVITY OF CARVACROL AGAINST SHRIMP PATHOGENS AND ITS USE AS FEED ADDITIVE FOR THE PACIFIC WHITE SHRIMP. Boletim do Instituto de Pesca, [S.l.], v. 47, sep. 2021. ISSN 1678-2305. Available at: <>. Date accessed: 26 may 2022. doi:
Scientific Article

Most read articles by the same author(s)