Clove Essential Oil from Eugenia caryophyllus Induces Anesthesia, Alters Swimming Performance, Heart Functioning and Decreases Survival Rate During Recovery of Daphnia magna

Clove oil is a common anesthetic used in aquaculture. However little is known on its effects on non-target crustaceans living in fish ponds and other natural reservoirs. The purpose of the investigation was to determine survival, time to anesthesia (TtA) and recovery (TtR), swimming activity, heart functioning during 5 and 45-min exposure of Daphnia magna to clove oil at concentrations of 150, 100, 50, 25, 10 and 1 µl/L. Swimming behavior and heart activity of Daphnia were evaluated by video image analysis with the use of Tracker® v.4.82 and Image Tool® software. The results showed that the oil induced anesthesia at three highest concentrations: 150, 100 and 50 μl/L. TtA and TtR were dependent on the size of the animal and concentration of the oil. The shortest TtA and the longest TtR were noted for the smallest daphnids anaesthetized for 45 minutes at 150, 100 and 50 ul/L. Lethality of small and large daphnids previously exposed to anesthetic concentrations for both times was observed at 24th hour of recovery period suggesting latent toxicity of clove oil. The oil inhibited heart rate, heart muscle contraction and swimming velocity at anesthetic (150, 100, 50 μl/L) and at non-anesthetic concentrations of 25, 10 and 1μl/L. Recovery period after previous 45-min anesthesia showed temporal increase however and subsequent decrease of Daphnia swimming velocity and heart activity resulting probably from latent toxicity. The results of the present study indicate that clove oil induces rapid immobilization and latent toxicity in Daphnia magna, at lower concentrations that those recommended for fish anesthesia and suggest that excessive use of the oil in natural ponds or other water reservoirs may be detrimental to coexisting cladocerans leading to disturbance of ecological balance.

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Ahmad, M.F. 1969. Anaesthetic effects of tricane methane sulphonate (MS 222 Sandoz) on Gammarus pulex (L.) (Amphipoda). Crustaceana, 16: 197201.

Akbari, S., Khoshnod, M.J., Rajaian, H. and Afsharnasab, M. 2010. The use of eugenol as an anesthetic in transportation of with indian shrimp (Fenneropenaeus indicus) post larvae. Turk. J. Fish. Aquat. Sci., 10: 423-429.

Akbulut, B., Cakmak, E., Aksungur, N. and Cavdar, Y. 2010. Effect of exposure duration on time to recovery from anaesthesia of clove oil in juvenile for Russian sturgeon. Turk. J. Fish. Aquat. Sci., 11(3):463-467. doi: 10.4194/1303-2712-v11_3_17.

Alqareer, A., Alyahya, A. and Andersson, L. 2006. The effect of clove and benzocaine versus placebo as topical anesthetics. J. Dent., 34:747-750.

American Society of Testing and Materials. 1986. Standard practice for conducting static acute toxicity tests on wastewaters with Daphnia: Annual book of ASTM standards. Vol. 11.04. Philadelphia: 4229-4284.

Anderson, W.G., McKinley, S.R. and Covalecchia, M. 1997. The use of clove oil as an anesthetic for rainbow trout and its effects on swimming performance. N. Am. J. Fish. Manage, 17: 301-307.

Arina, B. and Iqbal, A. 2002. In vitro fungitoxicity of the essential oil of Syzygium aromaticum. World J. Microbiol. Biotech., 18: 317-319.

Bekker, J.M. and Krijgsman, B.J. 1951. Physiological investigations into the heart function of Daphnia. J. Physiol., 115: 249-257.

Boyer, S.E., White, J.S., Stier, A.C. and Osenberg, C.W. 2009. Effects of the fish anesthetic, clove oil (eugenol), on coral health and growth. J. Exp. Mar. Biol. Ecol., 369: 5357.

Bownik, A., Stępniewska, Z. and Skowroński, T. 2014. Protective effects of ectoine on heat-stressed Daphnia magna. J. Comp. Physiol., 184(B): 961-976.

Bownik, A. and Stępniewska, Z. 2015. Protective effects of ectoine on behavioral, physiological and biochemical parameters of Daphnia magna subjected to hydrogen peroxide. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 170: 38-49 doi:10.1016/j.cbpc.2015.02.002

Campbell, A.K., Wann, K.T. and Matthews, S.B. 2004. Lactose causes heart arrhythmia in the water flea Daphnia pulex. Comp. Biochem. Physiol., 139(B): 225-234.

Chaieb, K., Zmantar, T., Ksouri, R., Hajlaoui, H., Mahdouani, K., Abdelly, C. and Bakhrouf, A. 2007. Antioxidant properties of essential oil of Eugenia caryophyllata and its antifungal activity against a large number of clinical Candida species. Mycosis, 50: 403-406.

Cooke, S.J., Suski, C.D., Ostrand, K.G., Tufts, B.L. and Wahl, D.H. 2004. Behavioral and physiological assessment of low concentrations of clove oil anaesthetic for handling and transporting largemouth bass (Micropterus salmoides). Aquaculture, 239: 509- 529.

Coyle, S.D., Durborow, R.M. and Tidwell, J.H. 2004. Anesthetics in aquaculture. SRAC Publication, No. 3900.

Coyle, S.D., Dasgupta, S., Tidwell, J.H., Beavers, T., Bright, L.A. and Yasharan, D.K. 2005. Comparative efficacy of anesthetics for the freshwater prawn Macrobrachium rosenbergii. J. World Aquacult. Soc., 36: 282290.

Damiani, C.E., Moreira, C.M., Zhang, H.T., Creazzo, T.L. and Vassallo, D.V. 2004. Effects of eugenol, an essential oil, on the mechanical and electrical activities of cardiac muscle. J. Cardiovasc. Pharmacol., 44:688-695.

Dodson, S.I. and Frey, DG. 2001. Cladocera and other branchiopoda. In: J.H. Thorp and A.P. Covich (Eds.), Ecology and classification of North American Freshwater Invertebrates. Elsevier, 778 pp.

FDA. 2007. Guidance for industry concerns related to the use of clove oil as an anesthetic for fish. Food and Drug Administration Center for Veterinary Medicine, Rockville, MD. www.fda.gov/downloads/animal veterinary/guidancecomplianceenforcement/guidancef orindustry/ucm052520.pdf

Fu, Y., Zu, Y., Chen, L., Shi, X., Wang, Z, Sun, S. and Efferth, T. 2007. Antimicrobial activity of clove and rosemary essential oils alone and in combination. Phytother. Res., 21: 989-994.

Hoskonen, P. and Pirhonen, J. 2004. Temperature effects on anaesthesia with clove oil in six temperate-zone fishes. J. Fish Biol., 64: 11361142.

Houston, A.H. and Corlett, J.T. 1976. Specimen weight and MS-222. J. Fish Res. Board Can., 33: 14031407.

Isaacs, G. 1983. Permanent local anaesthesia and anhydrosis after clove oil spillage. Lancet, 1: 882883.

Jaganathan, S.K., Mazumdar, A., Mondhe, D. and Mandal, M. 2011. Apoptotic effect of eugenol in human colon cancer cell lines. Cell Biol. Int., 35: 607615.

Lahlou, S., Interaminense, L.F., Magalhães, P.J., LealCardoso, J.H. and Duarte, G.P. 2004. Cardiovascular effects of eugenol, a phenolic compound present in many plant essential oils, in normotensive rats. J. Cardiovasc. Pharmacol., 43: 250 257. Munday, P.L. and Wilson, S.K. 1997. Comparative efficacy of clove oil and other chemicals in anesthetization of Pomacentrus amoboinensis, a coral reef fish. J. Fish Biol., 51: 931938.

Norton, J.H., Dashorst, M., Lansky, T.M. and Mayer, R.J. 1996. An evaluation of some relaxants for use with pearl oysters. Aquaculture, 144: 3952.

www.epa.gov/pesticides/biopesticides/ingredients/fact sheets/factsheet_florals.htm

Ögretmen, F. and Gӧkcek, K. 2013. Comparative Efficacy of three anesthetic agents on juvenile African Catfish, Clarias gariepinus (Burchell, 1822). Turk J Fish Aquat Sci., 13: 51-56.

Ozeki, M. 1975. The effects of eugenol on the nerve and muscle in crayfish. Comp. Biochem. Physiol. Part C, 50: 183-191.

Parodi, T.V., Cunha, M.A., Heldwein, C.G., de Souza, D.M., Martins, Á.C., Garcia, L.O., Wasielesky, WJr, Monserrat, J.M., Schmidt, D., Caron, B.O., Heinzmann, B. and Baldisserotto, B. 2012. The anesthetic efficacy of eugenol and the essential oils of Lippia alba and Aloysia triphylla in post-larvae and sub-adults of Litopenaeus vannamei (Crustacea, Penaeidae). Comp. Biochem. Physiol., 155(C):462 468.

Prashar, A., Locke, I.C. and Evans, C.S. 2006. Cytotoxicity of clove (Syzygium aromaticum) oil and its major components to human skin cells. Cell Prolif., 39: 241- 248.

Prince, A. and Powell, C. 2000. Clove oil as an anaesthetic for invasive field procedures on adult rainbow trout. N. Am. J. of Fish. Manag., 20: 10291032.

Saydmohammed, M. and Pal, A.K. 2009. Anesthetic effect of eugenol and menthol on handling stress in Macrobrachium rosenbergii. Aquaculture, 298: 162- 167.

Sensch, O., Vierling, W., Brandt, W. and Reiter, M. 2000. Effects of inhibition of calcium and potassium currents in guinea-pig cardiac contraction: comparison of b-caryophyllene oxide, eugenol, and nifedipine. Brit. J. Pharm., 131: 10891096.

Shima, J.S., Osenberg, C.W., Mary, C.M. and Rogers, L. 2006. Implications of changing coral communities: do larval traits or habitat features drive variation in density-dependent mortality and recruitment of juvenile reef fish? Proc. of the 10th Int. Coral Reef Symp., Okinawa, Japan: 226231.

Shimizu, N., Ogino, C., Kawanishi, T. and Hayashi, Y. 2002. Fractal analysis of Daphnia motion for acute toxicity bioassay. Inc. Environ. Toxicol., 17: 441-448.

Sladky, K.K., Swanson, C.R., Stoskopf, M.K., Loomis, 166 A. Bownik / Turk. J. Fish. Aquat. Sci. 15: 157-166 (2015) M.R. and Lewbart, G.A. 2001. Comparative efficacy of tricaine methanesulfonate and clove oil for use as anesthetics in red pacu (Piaractus brachypomis). Amer. J. Vet. Res., 62: 337342

Soto, C.G. 1995. Clove oil as a fish anaesthetic for measuring length and weight of rabbitfish (Siganus lineatus). Aquaculture, 136: 149152.

Svoboda, M. and Kolarova, J. 1999. A survey of anaesthetics used in the fish farming. Health Protection of Fish Proceeding of Papers. Research Institute of Fish Culture and Hydrobiology VodAany, 49-72. (In Czech):

Tierney, K.B. 2011. Behavioral assessments of neurotoxic effects and neurodegeneration in zebrafish. Biochim. Biophys. Acta (BBA) Mol. Basis Dis., 1812: 381- 389.

US EPA. 2004. Floral Attractants, Repellents, and Insecticides. US Environmental Protection Agency. Velisek, J., Svobodova, Z. and Piackova, V. 2005. Effects of clove oil anaesthesia on rainbow trout (Oncorhynchus mykiss). Acta Vet. Brno, 74: 139-146.

Venarsky, M.P. and Wilhelm, F.M. 2006. Use of clove oil to anaesthetize freshwater amphipods. Hydrobiologia, 568: 425-432.

Villegas-Navarro, A., Rosas, L.E. and Reyes, J.L. 2003. The heart of Daphnia magna: effects of four cardioactive drugs. Comp. Biochem. Physiol., 136(C): 127134.