Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2024-0107
Animal Reproduction (AR)
ORIGINAL ARTICLE

Tricaine methanesulfonate and eugenol during semen collection do not affect fertilization and hatching rates on Rhamdia quelen

Nathalia dos Santos Teixeira; Lis Santos Marques; Rômulo Batista Rodrigues; Thales de Souza França; Jhony Lisbôa Benato; Maritza Pérez Atehortúa; Darlan Gusso; Itamar Cossina Gomes; Thaiza Rodrigues de Freitas; Henrique Mautone Gomes; Helen Tais Rosa-Silva; Alexandre Kleber Silveira; José Claudio Fonseca Moreira; Danilo Pedro Streit Junior

http://dx.doi.org/10.1590/1984-3143-AR2024-0107 Anim Reprod, vol.23, n2, e20240107, 2026
PDF
Downloads: 0
Views: 94

Abstract

The reproduction of some fish species in captivity is only possible by imposing artificial reproductive procedures, and the manipulation of fish for these purposes is a stressor. Thus, anesthesia can reduce stress during handling. However, it is necessary to investigate the possible side effects on breeding and the general health of the animal. Therefore, we aimed to investigate the impact of tricaine methanesulfonate (MS-222) and eugenol (EUG) at concentrations of 300 mg L-1 and 40 mg L-1, respectively, on several stress- and reproductive-related parameters in Rhamdia quelen anesthetized before semen collection. After hormonal induction, 24 sexually mature males (534.4 ± 259 g) were randomly assigned to treatment groups and semen and blood samples were collected. Anesthesia recovery and induction time were extended in fish anesthetized with EUG; however, plasma cortisol concentrations did not differ among the treatment groups. EUG caused higher DNA fragmentation in blood cells than in the control group (without anesthesia). EUG increased the monocyte count compared to the other experimental groups. MS-222 showed a lower sulfhydryl group (SH) quantitation than the other experimental groups. The anesthetics used before semen collection in this study had no adverse effects on the fertilization or hatching capacity of anesthetized R. quelen. MS-222 (300 mg L-1) and EUG (40 mg L-1), despite being related to hematological and semen changes, did not negatively affect the reproductive capacity of R. quelen males. Thus, anesthesia is recommended for R. quelen during semen collection, considering reproductive parameters and animal welfare.

Keywords

DNA damage, fish anesthesia, fish welfare, oxidative stress, reproductive management

References

Allison LN. The effect of Tricaine Methanesulfonate (M.S. 222) on the motility of brook trout sperm. Prog Fish-Cult. 1961;23(1):46-7. https://doi.org/10.1577/1548-8659(1961)23[46:TEOTMM]2.0.CO;2.

Amado LL, Garcia ML, Ramos PB, Freitas RF, Zafalon B, Ferreira JLR, Yunes JS, Monserrat JM. A method to measure total antioxidant capacity against peroxyl radicals in aquatic organisms: application to evaluate microcystins toxicity. Sci Total Environ. 2009;407(6):2115-23. https://doi.org/10.1016/j.scitotenv.2008.11.038. PMid:19095287.

Aravena-Canales D, Valenzuela-Muñoz V, Gallardo-Escarate C, Molina A, Valdés JA. Molecular and epigenetic responses to crowding stress in rainbow trout (Oncorhynchus mykiss) skeletal muscle. Front Endocrinol. 2025;16:1571111. https://doi.org/10.3389/fendo.2025.1571111. PMid:40309435.

Ayala-Soldado N, Mora-Medina R, Molina-López AM, Lora-Benítez AJ, Moyano-Salvago R. Evaluation of the effectiveness of eugenol and MS-222 as anesthetics in zebrafish in repeated exposures and post-anesthesia behaviour. Animals. 2024;14(16):2418. https://doi.org/10.3390/ani14162418. PMid:39199952.

Ayyat MS, Khalil NA, Al-Sagheer AA, Elmemy MM, Abdel-Monem UM. Maintaining the physiological stability during artificial spawning of Liza ramada in captivity. Aquacult Int. 2025;33(1):44. https://doi.org/10.1007/s10499-024-01721-y.

Azizi S, Balasch JC, Cartan S, Jerez-Cepa I, Mancera JM, Tort L, Khansari AR. Enhancing farmed fish welfare: evaluating the effectiveness of plant-based stress mitigating agents as sedatives in sea bass (Dicentrarchus labrax) following intraperitoneal vaccination. Fish Shellfish Immunol. 2025;156:110058. https://doi.org/10.1016/j.fsi.2024.110058. PMid:39613169.

Bagheri S, Gholamhosseini A, Yaghoobpour T, Adel M. Uses of Eugenol in aquaculture: benefits and applications: a review. Ann Anim Sci. 2025;25(3):945-95. https://doi.org/10.2478/aoas-2024-0109.

Barbas LAL, Torres MF, Costa BMP, Feitosa MJM, Maltez LC, Amado LL, Toda YPS, Batista PS, Cabral DAC, Hamoy M. Eugenol induces body immobilization yet evoking an increased neuronal excitability in fish during short-term baths. Aquat Toxicol. 2021;231:105734. https://doi.org/10.1016/j.aquatox.2020.105734. PMid:33385846.

Barcellos G, Kreutz LC, Souza C, Rodrigues LB, Fioreze I, Quevedo RM et al. Hematological changes in jundiá, Rhamdia quelen (Quoy and Gaimard Pimelodidae), after acute and chronic stress caused by usual aquacultural management, with emphasis on immunosuppressive effects. Aquaculture. 2004;237(1-4):229-36. https://doi.org/10.1016/j.aquaculture.2004.03.026.

Barcellos LJG, Woehl VM, Wassermann GF, Quevedo RM, Ittzés I, Krieger MH. Plasma levels of cortisol and glucose in response to capture and tank transference in Rhamdia quelen (Quoy & Gaimard), a South American catfish. Aquacult Res. 2001;121–123(2):121-3. https://doi.org/10.1046/j.1365-2109.2001.00539.x.

Barton BA, Iwama GK. Physiological changes in fish from stress in aquaculture with emphasis on the response and effects of corticosteroids. Annu Rev Fish Dis. 1991;1:3-26. https://doi.org/10.1016/0959-8030(91)90019-G.

Blom E. A one-minute live-dead sperm stain by means of eosin-nigrosin. Fertil Steril. 1950;1(2):176-7. https://doi.org/10.1016/S0015-0282(16)30125-X.

Boaventura TP, Souza CF, Ferreira AL, Favero GC, Baldissera MD, Heinzmann BM, Baldisserotto B, Luz RK. Essential oil of Ocimum gratissimum (Linnaeus, 1753) as anesthetic for Lophiosilurus alexandri: Induction, recovery, hematology, biochemistry and oxidative stress. Aquaculture. 2020;529:735676. https://doi.org/10.1016/j.aquaculture.2020.735676.

Bojarski B, Witeska M, Kondera E. Blood biochemical biomarkers in fish toxicology-a review. Animals (Basel). 2025;15(7):965. https://doi.org/10.3390/ani15070965. PMid:40218358.

Bombardelli RA, Morschbacher EF, Campagnolo R, Sanches EA, Syperreck MA. Insemination dose for artificial fertilization of grey jundia oocytes, Rhamdia quelen (Quoy & Gaimardm, 1824). Rev Bras Zootec. 2006;35:1251-7. https://doi.org/10.1590/S1516-35982006000500001.

Cabrita E, Martínez-Páramo S, Gavaia PJ, Riesco MF, Valcarce DG, Sarasquete C, Herráez MP, Robles V. Factors enhancing fish sperm quality and emerging tools for sperm analysis. Aquaculture. 2014;432:389-401. https://doi.org/10.1016/j.aquaculture.2014.04.034.

Campbell DLM, Lea JM, Keshavarzi H, Lee C. Virtual fencing is comparable to electric tape fencing for cattle behavior and welfare. Front Vet Sci. 2019;6:445. https://doi.org/10.3389/fvets.2019.00445. PMid:31921906.

Carolsfeld J, Godinho HP, Zaniboni Filho E, Harvey BJ. Cryopreservation of sperm in Brazilian migratory fish conservation. J Fish Biol. 2003;63(2):472-89. https://doi.org/10.1046/j.1095-8649.2003.00170.x.

Cericato L, Neto JGM, Kreutz LC, Quevedo RM, da Rosa JGS, Koakoski G, Centenaro L, Pottker E, Marqueze A, Barcellos LJ. Responsiveness of the interrenal tissue of Jundiá (Rhamdia quelen) to an in vivo ACTH test following acute exposure to sublethal concentrations of agrichemicals. Comp Biochem Physiol C Toxicol Pharmacol. 2009;149(3):363-7. https://doi.org/10.1016/j.cbpc.2008.09.002. PMid:18824247.

Coimbra VC, Rodrigues J, Santos RS, Rodrigues RB, Streit-Jr D, Caldas ALS, Albuquerque ESN, Ferreira EJS, Maximino C, Siqueira-Silva DH. Cryopreserved sperm does not affect larval ontogeny and quality in Rhamdia quelen. Fish Physiol Biochem. 2025;51(1):42. https://doi.org/10.1007/s10695-025-01455-5. PMid:39907872.

Collins AR. The comet assay: a heavenly method! Mutagenesis. 2015;30(1):1-4. https://doi.org/10.1093/mutage/geu079. PMid:25527721.

Collymore C, Banks EK, Turner PV. Lidocaine hydrochloride compared with MS222 for the euthanasia of zebrafish (Danio rerio). J Am Assoc Lab Anim Sci. 2016;55(6):816-20. PMid:27931323.

Corso MN, Marques LS, Gracia LFG, Rodrigues RB, Barcellos LJG, Streit DP Jr. Effects of different doses of eugenol on plasma cortisol levels and the quality of fresh and frozen-thawed sperm in South American catfish (Rhamdia quelen). Theriogenology. 2019;125:135-9. https://doi.org/10.1016/j.theriogenology.2018.10.033. PMid:30414567.

Dhabhar FS. Effects of stress on immune function: the good, the bad, and the beautiful. Immunol Res. 2014;58(2-3):193-210. https://doi.org/10.1007/s12026-014-8517-0. PMid:24798553.

Dietrich GJ, Kowalski R, Wojtczak M, Dobosz S, Goryczko K, Ciereszko A. Motility parameters of rainbow trout (Oncorhynchus mykiss) spermatozoa in relation to sequential collection of milt, time of post-mortem storage and anesthesia. Fish Physiol Biochem. 2006;31(1):1-9. https://doi.org/10.1007/s10695-005-3527-4.

Dong H, Wang W, Duan Y, Li H, Liu Q, Sun Y, Zhang J. Transcriptomic analysis of juvenile Chinese sea bass (Lateolabrax maculatus) anesthetized by MS-222 (tricaine methanesulfonate) and eugenol. Fish Physiol Biochem. 2020;46(3):909-20. https://doi.org/10.1007/s10695-019-00755-x. PMid:31916052.

Ellman GL. Tissue sufhydryl groups. Arch Biochem Biophys. 1959;82(1):70-7. https://doi.org/10.1016/0003-9861(59)90090-6. PMid:13650640.

Esterbauer H, Cheeseman KH. Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods Enzymol. 1990;186:407-21. https://doi.org/10.1016/0076-6879(90)86134-H. PMid:2233308.

Félix L, Vieira R, Monteiro SM, Venâncio C. A meta‐analytic review of monoterpene for fish anaesthesia. Fish Fish. 2023;24(3):367-80. https://doi.org/10.1111/faf.12731.

Ferreira JM, Jorge S, Félix L, Morello GM, Olsson IAS, Valentim AM. Behavioural aversion and cortisol level assessment when adult zebrafish are exposed to different anaesthetics. Biology (Basel). 2022;11(10):1433. https://doi.org/10.3390/biology11101433. PMid:36290337.

Fish RE, Brown MJ, Danneman PJ, Karas AZ. Anesthesia and analgesia in laboratory animals, laboratory animal health services. 2nd ed. Amsterdam: Elsevier; 2008.

Fortes CHM, Ferrari FT, Baldisserotto B, Schmidt D, Sutili FJ, Heiznmann BM. anesthetic potential of essential oils from brazilian native plants in Rhamdia quelen juveniles (silver catfish). Neotrop Ichthyol. 2024;22(3):e240034. https://doi.org/10.1590/1982-0224-2024-0034.

França TS, Gomes IC, Sanches EA, Pérez-Atehortúa MP, Teixeira NS, Rodrigues RB et al. Fish sperm cryopreservation using biodegradable containers: new low-cost and environment-friendly methodology. Reproduction. 2023;166(2):89-97. https://doi.org/10.1530/REP-23-0046. PMid:37204211.

Goes MD, Goes ESR, Ribeiro RP, Lopera-Barrero NM, Castro PL, Bignotto TS et al. Natural and artificial spawning strategies with fresh and cryopreserved semen in Rhamdia quelen: reproductive parameters and genetic variability of offspring. Theriogenology. 2017;88:254-63.e3. https://doi.org/10.1016/j.theriogenology.2016.09.029. PMid:27814913.

Gomes DP, Chaves BW, Becker AG, Baldisserotto B. Water parameters affect anaesthesia induced by eugenol in silver catfish, Rhamdia quelen. Aquacult Res. 2011;2011(6):878-86. https://doi.org/10.1111/j.1365-2109.2011.02864.x.

Gorissen M, Flik G. The endocrinology of the stress an Adaptation-Physiological View. In: Schreck CS, Tort L, Farrell AP, Brauner CJ, editors. Fish physiology. Cambridge, MA: Academic Press; 2016. p. 75-111. (Vol. 35). https://doi.org/10.1016/B978-0-12-802728-8.00003-5.

Gouveia K, Hurst JL. Improving the practicality of using non-aversive handling methods to reduce background stress and anxiety in laboratory mice. Sci Rep. 2019;9(1):20305. https://doi.org/10.1038/s41598-019-56860-7. PMid:31889107.

Gressler LT, Parodi TV, Riffel APK, DaCosta ST, Baldisserotto B. Immersion anaesthesia with tricaine methanesulphonate or propofol on different sizes and strains of silver catfish Rhamdia quelen. J Fish Biol. 2012;81(4):1436-45. https://doi.org/10.1111/j.1095-8649.2012.03409.x. PMid:22957883.

Gressler LT, Riffel APK, Parodi TV, Saccol EMH, Koakoski G, Da Costa ST, Pavanato MA, Heinzmann BM, Caron B, Schmidt D, Llesuy SF, Barcellos LJG, Baldisserotto B. Silver catfish Rhamdia quelen immersion anaesthesia with essential oil of Aloysia triphylla (L’Hérit) Britton or tricaine methanesulfonate: effect on stress response and antioxidant status. Aquacult Res. 2014;45(6):1061-72. https://doi.org/10.1111/are.12043.

Gressler LT, Sutili FJ, Loebens L, Medianeira E, Baldisserotto B. Histological and antioxidant responses in Rhamdia quelen sedated with propofol. Aquacult Res. 2015;47(7):2297-306. https://doi.org/10.1111/are.12682.

Guptha JS, Renuka M, Suneetha Y, Reddy MS. Evaluation of antioxidant defence system during xenobiotic induced oxidative stress in freshwater fish Oreochromis mossambicus. Int J Fish Aquat Sci. 2016;4:379-85.

He R, Lei B, Su Y, Wang A, Cui K, Shi X, Chen X. Effectiveness of eugenol as an anesthetic for adult spotted sea bass (Lateolabrax maculatus). Aquaculture. 2020;523:735180. https://doi.org/10.1016/j.aquaculture.2020.735180.

Hilbig CC, do Nascimento NF, Heinen AL, Neto AT, Funghetto JP, Bombardelli RA et al. Effects of dietary fatty acids on the reproduction of South American female catfish Rhamdia quelen (Quoy & Gaimard, 1824). Lat Am J Aquat Res. 2019;47(3):456-66. https://doi.org/10.3856/vol47-issue3-fulltext-8.

Hoseini SM, Taheri Mirghaed A, Yousefi M. Application of herbal anaesthetics in aquaculture. Rev Aquacult. 2018;11(3):550-64. https://doi.org/10.1111/raq.12245.

Hsu JCN, Rairat T, Lu YP, Chou CC. The use of tricaine methanesulfonate (MS-222) in Asian seabass (Lates calcarifer) at different temperatures: study of optimal doses, minimum effective concentration, blood biochemistry, immersion pharmacokinetics, and tissue distributions. Vet Sci. 2023;10(9):539. https://doi.org/10.3390/vetsci10090539. PMid:37756061.

Jia Y, Xie T, Gao Y, Qin H, Guan C. Anesthetics efficacy and physiological response of MS222 and clove oil in spotted knifejaw Oplegnathus punctatus. Aquacult Rep. 2022;25:101201. https://doi.org/10.1016/j.aqrep.2022.101201.

Junmahasathien T, Kheawfu K, Chittasupho C, Nuanmanee S, Chaisri W, Pikulkaew S. An evaluation of the anesthetic properties and serum biochemical response of Ocimum essential oil as a new anesthetic agent for goldfish (Carassius auratus). Vet Sci. 2025;12(11):1069. https://doi.org/10.3390/vetsci12111069. PMid:41295707.

Kamatou GP, Vermaak I, Viljoen AM. Eugenol - From the remote Maluku Islands to the international market place: a review of a remarkable and versatile molecule. Molecules. 2012;17(6):6953-81. https://doi.org/10.3390/molecules17066953. PMid:22728369.

Klak K, Maciuszek M, Pijanowski L, Marcinkowska M, Homa J, Verburg-van Kemenade BML, Rakus K, Chadzinska M. Evolutionarily conserved mechanisms regulating stress-induced neutrophil redistribution in fish. Front Immunol. 2024;15:1330995. https://doi.org/10.3389/fimmu.2024.1330995. PMid:38515741.

Lavalle C, Clarence S, Khan H, Shires K, Parrott J. Current research and guidelines for euthanasia in laboratory fish with a focus on fathead minnows. Lab Anim. 2025;59(3):319-31. https://doi.org/10.1177/00236772241288146. PMid:39862184.

Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz A, Ahn BW, Shaltiel S, Stadtman ER. Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol. 1990;186:464-78. https://doi.org/10.1016/0076-6879(90)86141-H. PMid:1978225.

Li X, Lin X, Zhuang Z, Luo Y, Li Y, Isse SA, Chen Z, Meng F, Luo Q, Zhu B, Chen X. Reproductive toxicity and transgenerational effects of anesthetic etomidate in zebrafish: PCOS-like ovarian changes, oxidative stress, and metabolism disruption. Environ Health. 2025;3(11):1353-65. https://doi.org/10.1021/envhealth.5c00156. PMid:41306307.

Macaulay G, Bui S, Oppedal F, Dempster T. Challenges and benefits of applying fish behaviour to improve production and welfare in industrial aquaculture. Rev Aquacult. 2020;13(2):934-48. https://doi.org/10.1111/raq.12505.

Maricchiolo G, Genovese L. Some contributions to knowledge of stress response in innovative species with particular focus on the use of the anaesthetics. Open Mar Biol J. 2011;5(1):24-33. https://doi.org/10.2174/1874450801105010024.

Martins C, Rueff J, Rodrigues AS. Genotoxic alkenylbenzene flavourings, a contribution to risk assessment. Food Chem Toxicol. 2018;118:861-79. https://doi.org/10.1016/j.fct.2018.06.020. PMid:29913232.

Miliorini AB, Murgas LDS, Rosa PV, Oberlender G, Pereira GJM, Da Costa DV. A morphological classification proposal for curimba (Prochilodus lineatus) sperm damages after cryopreservation. Aquacult Res. 2011;42(2):177-87. https://doi.org/10.1111/j.1365-2109.2010.02575.x.

Minaz M, Alparslan C, Er A. Using machine learning to predict anesthetic dose in fish: a case study using nutmeg oil. Front Vet Sci. 2025;12:1652115. https://doi.org/10.3389/fvets.2025.1652115. PMid:40927168.

Minaz M, Félix L. Trends and advancements in fish anaesthesia over the last decade. Aquaculture. 2025;612:743147. https://doi.org/10.1016/j.aquaculture.2025.743147.

Nascimento HS, Crispim BA, Francisco LFV, Merey FM, Kummrow F, Viana LF, Inoue LAKA, Barufatti A. Genotoxicity evaluation of three anesthetics commonly employed in aquaculture using Oreochromis niloticus and Astyanax lacustris. Aquacult Rep. 2020;17:100357. https://doi.org/10.1016/j.aqrep.2020.100357.

Nascimento TP, Barros FJT, Couto OLP, Paz MDR, Junior SVN, Sousa LM, Pereira TS. Anesthesia of Baryancistrus xanthellus (Siluriformes: Loricariidae): bioassays with exposure and re-exposure cause reverse tolerance to eugenol. Aquacult Rep. 2025;42:102807. https://doi.org/10.1016/j.aqrep.2025.102807.

Neumann G, Sanches PV, Bombardelli RA. Effects on fertility of motile sperm to egg ratio with use of cryopreserved Rhamdia quelen semen at different post-activation times. Anim Reprod Sci. 2019;201:84-92. https://doi.org/10.1016/j.anireprosci.2019.01.001. PMid:30621964.

Ohnesorge N, Heinl C, Lewejohann L. Current methods to investigate nociception and pain in zebrafish. Front Neurosci. 2021;15:632634. https://doi.org/10.3389/fnins.2021.632634. PMid:33897350.

Ohnesorge N, Wilzopolski J, Steinfath M, Lewejohann L, Banneke S, Heinl C. Assessment of the effect of tricaine (MS-222)-induced anesthesia on brain-wide neuronal activity of zebrafish (Danio rerio) larvae. Front Neurosci. 2024;18:1456322. https://doi.org/10.3389/fnins.2024.1456322. PMid:39381680.

Oliveira CB, Lemos CHP, Vidal LVO, Couto RD, Pereira DSP, Copatti CE. Anaesthesia with eugenol in hybrid Amazon catfish (Pseudoplatystoma reticulatum × Leiarius marmoratus) handling: biochemical and haematological responses. Aquaculture. 2019a;501:255-9. https://doi.org/10.1016/j.aquaculture.2018.11.046.

Oliveira RH, Pereira‐da‐Silva EM, Viegas EMM. Clove oil attenuates stress responses in lambari, Astyanax altiparanae. Aquacult Res. 2019b;2019:1-7. https://doi.org/10.1111/are.14293.

Pereira CR, Barcellos LJG, Kreutz LC, Quevedo RM, Ritter F, Silva LB. Embryonic and larval development of jundiaá (Rhamdia quelen, Quoy & Gaimard, 1824, Pisces, Teleostei), a South American catfish. Braz J Biol. 2006;66(4):1057-63. https://doi.org/10.1590/S1519-69842006000600013. PMid:17299942.

Pérez-Atehortúa M, Galuppo AG, Rodrigues RB, França TS, Teixeira NS, Freitas TR et al. The use of differential separation and density gradient with AllGrad 90% after thawing improves the sperm quality of South American catfish (Rhamdia quelen). Aquaculture. 2022;553:738072. https://doi.org/10.1016/j.aquaculture.2022.738072.

Pickering AD, Pottinger TG, Christie P. Recovery of the brown trout, Salmo trutta L, from acute handling stress : a time-course study. J Fish Biol. 1982;20(2):229-44. https://doi.org/10.1111/j.1095-8649.1982.tb03923.x.

Pietsch C, Pawlak P, Konrad J. Acute stress effects over time on the stress axis in the carp (Cyprinus carpio) brain. Aquacult Res. 2025;2025(1):121460. https://doi.org/10.1155/are/1121460.

Popovic BNT, Strunjak-Perovic I, Coz-Rakovac R, Barisic J, Jadan M, Berakovic AP, Klobucar RS. Tricaine methane-sulfonate (MS-222) application in fish anaesthesia. J Appl Ichthyology. 2012;2012:1-12. https://doi.org/10.1111/j.1439-0426.2012.01950.x.

Ranzani-Paiva MJT, Pádua SB, Tavares-Dias M, Egami MI. Métodos para análise hematológica em peixes. 1. ed. Maringá: SciELO/EDUEM; 2013. https://doi.org/10.7476/9788576286530.

Readman GD, Owen SF, Knowles TG, Murrell JC. Species specific anaesthetics for fish anaesthesia and euthanasia. Sci Rep. 2017;7(1):7102. https://doi.org/10.1038/s41598-017-06917-2. PMid:28769117.

Rosa-Silva HT, Panzenhagen AC, Espitia-Pérez P, Teixeira AA, Roitman A, Almeida RF et al. Effects of foetal and breastfeeding exposure to methylmercury (MeHg) and retinol palmitate (Vitamin A) in rats: redox parameters and susceptibility to DNA damage in liver. Mutat Res Genet Toxicol Environ Mutagen. 2020;858-860:503239. https://doi.org/10.1016/j.mrgentox.2020.503239. PMid:33198929.

Ross L, Ross B. Anaesthetic and Sedative Techniques for Aquatic Animals. 3rd ed. Oxford: Wiley-Blackwell; 2008.. https://doi.org/10.1002/9781444302264.

Salah A, Bouaziz C, Amara I, Abid-Essefi S, Bacha H. Eugenol protects against citrinin-induced cytotoxicity and oxidative damages in cultured human colorectal HCT116 cells. Environ Sci Pollut Res Int. 2019;26(30):31374-83. https://doi.org/10.1007/s11356-019-06212-9. PMid:31473926.

Salbego J, Toni C, Becker AG, Zeppenfeld CC, Menezes CC, Loro VL, Heinzmann BM, Baldisserotto B. Biochemical parameters of silver catfish (Rhamdia quelen) after transport with eugenol or essential oil of Lippia alba added to the water. Braz J Biol. 2017;77(4):696-702. https://doi.org/10.1590/1519-6984.16515. PMid:28492807.

Sanches EA, Neumann G, Toledo CPR, Bombardelli RA, Piana PA, Romagosa E. Temperature and storage period over spermatic parameters of jundiá,Rhamdia quelen (Quoy & Gaimard, 1824). Aquacult Res. 2011;44(4):534-41. https://doi.org/10.1111/j.1365-2109.2011.03056.x.

Sandoval‐Vargas L, Jiménez MS, González JR, Villalobos EF, Cabrita E, Isler IV. Oxidative stress and use of antioxidants in fish semen cryopreservation. Rev Aquacult. 2020;13(1):365-87. https://doi.org/10.1111/raq.12479.

Schreck CB, Tort L. 2016. Biology of Stress in Fish. In: Schreck CB, Tort L, Farrell AP, Brauner CJ, editors. The Concept of Stress in Fish. 1st ed. London: Elsevier; 2016.

Seibel H, Baßmann B, Rebl A. Blood will tell: what hematological analyses can reveal about fish welfare. Front Vet Sci. 2021;8:616955. https://doi.org/10.3389/fvets.2021.616955. PMid:33860003.

Silva LDL, Silva DT, Garlet QI, Cunha MA, Mallmann CA, Baldisserotto B et al. Anesthetic activity of Brazilian native plants in silver catfish (Rhamdia quelen). Neotrop Ichthyol. 2013;11(2):443-51. https://doi.org/10.1590/S1679-62252013000200014.

Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 1988;175(1):184-91. https://doi.org/10.1016/0014-4827(88)90265-0. PMid:3345800.

Sneddon LU. Clinical anesthesia and analgesia in fish. J Exot Pet Med. 2012;21(1):32-43. https://doi.org/10.1053/j.jepm.2011.11.009.

Sneddon LU. Evolution of nociception and pain: evidence from fish models. Philos Trans R Soc Lond B Biol Sci. 2019;374(1785):20190290. https://doi.org/10.1098/rstb.2019.0290. PMid:31544617.

Sopinka NM, Donaldson MR, O’Connor CM, Suski CD, Cooke SJ. Stress indicators in fish. 1st ed. In Schreck CB, Tort L, Farrell AP, Brauner CJ, editors. The Concept of Stress in Fish, London: Elsevier; 2016.. https://doi.org/10.1016/B978-0-12-802728-8.00011-4.

Souza VV, Moreira DP, Braz-Mota S, Valente W, Cotta GC, Rodrigues MS, Nóbrega RH, Corrêa RDS, Hoyos DCM, Sanches EA, Val AL, Lacerda SMDSN. Simulated climate change and atrazine contamination can synergistically impair zebrafish testicular function. Sci Total Environ. 2024;946:174173. https://doi.org/10.1016/j.scitotenv.2024.174173. PMid:38925398.

Streit-Junior D, Moraes G, Ribeiro R, Povh J, Souza E. Avaliação de diferentes técnicas para coloração de sêmen de peixes. Arq Ciências Veterinárias e Zool da UNIPAR. 2004;7:157-62.

Tago A, Yokoyama S, Ishikawa M, Koshio S. Pharmacokinetics of Eugenol in Japanese Flounder, Paralichthys olivaceus. J World Aquacult Soc. 2017;49(4):780-7. https://doi.org/10.1111/jwas.12438.

Teixeira NS, Marques LS, Rodrigues RB, Gusso D, Pinheiro GT, Machado TLF, Streit DP Jr. Effects of anesthetic MS-222 on stress and reproduction of South American silver catfish (Rhamdia quelen) males. Anim Reprod Sci. 2021;225:106669. https://doi.org/10.1016/j.anireprosci.2020.106669. PMid:33360163.

Tort L, Puigcerver M, Crespo S, Padrós F. Cortisol and haematological response in sea bream and trout subjected to the anaesthetics clove oil and 2-phenoxyethanol. Aquacult Res. 2002;33(11):907-10. https://doi.org/10.1046/j.1365-2109.2002.00741.x.

Velisek J, Stara A, Li Z, Silovska S, Turek J. Comparison of the effects of four anaesthetics on blood biochemical pro fi les and oxidative stress biomarkers in rainbow trout. Aquaculture. 2011;310(3-4):369-75. https://doi.org/10.1016/j.aquaculture.2010.11.010.

Wagner E, Arndt R, Hilton B. Physiological stress responses, egg survival and sperm motility for rainbow trout broodstock anesthetized with clove oil, tricaine methanesulfonate or carbon dioxide. Aquaculture. 2002;211(1-4):353-66. https://doi.org/10.1016/S0044-8486(01)00878-X.

Wojan EM, Carreiro NC, Clendenen DA, Neldner HM, Castillo C, Bertram SM, Kolluru GR. The effects of commonly used anaesthetics on colour measurements across body regions in the poeciliid fish, Girardinus metallicus. J Fish Biol. 2019;95(5):1320-30. https://doi.org/10.1111/jfb.14138. PMid:31515796.

Wong D, von Keyserlingk MAG, Richards JG, Weary DM. Conditioned place avoidance of zebrafish (Danio rerio) to three chemicals used for euthanasia and anesthesia. PLoS One. 2014;9(2):e88030. https://doi.org/10.1371/journal.pone.0088030. PMid:24505365.

Yada T, Tort L. Biology of stress in fish. In: Schreck CB, Tort L, Farrell AP, Brauner CJ, editors. Stress and disease resistance: immune system and immunoendocrine interactions. 1st ed. London: Elsevier Inc.; 2016, p. 365-403.

Yogalakshmi B, Viswanathan P, Anuradha CV. Investigation of antioxidant, anti-inflammatory and DNA-protective properties of eugenol in thioacetamide-induced liver injury in rats. Toxicology. 2010;268(3):204-12. https://doi.org/10.1016/j.tox.2009.12.018. PMid:20036707.

Zahran E, Risha E, Rizk A. Comparison propofol and eugenol anesthetics efficacy and effects on general health in Nile Tilapia. Aquaculture. 2021;534:736251. https://doi.org/10.1016/j.aquaculture.2020.736251.

Zanin M, Varela AS Jr, Acosta IB, Gheller SMM, Zimerman E, Froes CN et al. Tricaine methanesulfonate (MS-222) on the spermatic quality of zebrafish, Danio rerio. Aquaculture. 2021;533:736090. https://doi.org/10.1016/j.aquaculture.2020.736090.
 

Submitted date:
09/11/2024

Accepted date:
12/17/2025

69c44aaaa953951933112684 animreprod Articles
Links & Downloads
1984-3143 (Electronic) 1806-9614 (Printed)
Anim Reprod ©2026 All rights reserved.

Anim Reprod

Share this page
Page Sections