ABSTRACT
The aim of this study was to determine the anesthetic efficacy of the essential oils (EOs) of Aniba rosaeodora (EOAR) and Aniba parviflora (EOAP) and one of their main compounds, linalool, in two forms: synthetic and extracted from EOAR (linalool-AR) in tambaqui (Colossoma macropomum). In the first experiment, the anesthetic induction and recovery of juveniles exposed to 25- 200 µL L-1 of EOAR or 50- 300 µL L-1 of EOAP or synthetic linalool or linalool-AR was evaluated. The second experiment observed the behavioral effects of long-term exposure (12h) of these EOs and linalools (5 and 10 µL L-1). Fish exposed to 50-200 µL L-1 of EOAR and 100-300 µL L-1 of EOAP and both linalools reached deep anesthesia between 1-10 min. Induction time for all anesthesia stages decreased with the increasing concentration of the anesthetics. Linalool-AR showed lengthier time for anesthesia induction in some stages and for recovery at 100 and 200 µL L-1 in comparison to synthetic linalool. Normal equilibrium and swimming behavior was observed in fish exposed to the EOs and linalools throughout the 12 h of exposure. In conclusion, both EOs and linalools can be used as anesthetics and sedatives in tambaqui.
Keywords:
Anesthetics; Linalool isomers; Rosewood oil; Sedation; Swimming behavior
RESUMO
O objetivo deste trabalho foi determinar a eficácia anestésica dos óleos essenciais (OEs) de Aniba rosaeodora (OEAR) Aniba parviflora (OEAP) e um de seus compostos majoritários, linalol, em duas formas: sintética e extraída a partir de OEAR (linalol-AR) em tambaqui (Colossoma macropomum). No primeiro experimento avaliou-se a indução anestésica e a recuperação de juvenis expostos a 25- 200 µL L-1 de EOAR ou 50- 300 µL L-1 de EOAP ou linalol sintético ou linalol-AR. No segundo experimento observaram-se os efeitos comportamentais de uma longa exposição (12h) a estes OEs e linalois (5 e 10 µL L-1). Os peixes expostos a 50-200 µL L-1 de EOAR e 100-300 µL L-1 de OEAP e ambos os linalois alcançaram anestesia profunda entre 1-10 min. Tempo de indução a todos os estágios de anestesia diminuiu com o aumento na concentração dos anestésicos. Linalol-AR levou um maior tempo para induzir anestesia e para recuperação com 100 e 200 µL L-1 em comparação ao composto sintético. Peixes expostos aos OES e linalois por 12 h apresentaram equilíbrio e comportamento natatório normais. Em conclusão, tanto os OEs como o linalol sintético ou natural de AR podem ser usados como anestésicos e sedativos em tambaqui.
Palavras-chave:
Anestésicos; Isômeros de linalool; Natação; Óleo de palisandro; Sedação
Introduction
The anesthetic efficacy of several vegetable extractives and essential oils (EO) have been demonstrated in fish (Keene et al., 1988Keene JL, Noakes DLG, Moccia RD, Soto CG. The efficacy of clove oil as an anaesthetic for rainbow trout, Oncorhynchus mykiss (Walbaum). Aquac Res . 1988; 29(2):89-101.; Cunha et al., 2010Cunha MA, Barros FMC, Garcia LO, Veeck APL, Heinzmann BM, Loro VL et al. Essential oil of Lippia alba: A new anesthetic for silver catfish, Rhamdia quelen. Aquaculture . 2010; 306:403-6.; Pádua et al., 2013Pádua SB, Dias Neto J, Sakabe R, Claudiano GS, Chagas EC, Pilarski F. Variáveis hematológicas em tambaquis anestesiados com óleo de cravo e benzocaína. Pesqui Agropec Bras. 2013; 48(8):1171-74.; Silva et al., 2013aSilva LL, Garlet QI, Benovit SC, Dolci G, Mallmann CA, Bürger ME et al. Sedative and anesthetic activities of the essential oils of Hyptis mutabilis (Rich.) Briq. and their isolated components in silver catfish (Rhamdia quelen). Braz J Med Biol Res . 2013a; 46(9):771-09., bSilva LL, 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 . 2013b; 11:443-51.; Parodi et al., 2014Parodi TV, Cunha MA, Becker AG, Zeppenfeld CC, Martins DI, Koakoski G et al. Anesthetic activity of the essential oil of Aloysia triphylla and effectiveness in reducing stress during transport of albino and gray strains of silver catfish, Rhamdia quelen. Fish Physiol Biochem . 2014; 40(2):323-34.; Benovit et al., 2015Benovit SC, Silva LL, Salbego J, Loro VL, Mallmann CA, Baldisserotto B, Flores EM, Heinzmann BM. Anesthetic activity and bio-guided fractionation of the essential oil of Aloysia gratissima (Gillies & Hook.) Tronc. in silver catfish Rhamdia quelen. An Acad Bras Ciênc. 2015; 87(3):1675-89.; Barbas et al., 2016Barbas LAL, Stringhetta GR, Garcia LO, Figueiredo MRC, Sampaio LA. Jambu, Spilanthes acmella as a novel anaesthetic for juvenile tambaqui, Colossoma macropomum: Secondary stress responses during recovery. Aquaculture . 2016; 456:70-05., 2017Barbas LAL, Maltez LC, Stringhetta GR, Garcia LO, Monserrat JM, Silva DT et al. Properties of two plant extractives as anaesthetics and antioxidants for juvenile tambaqui Colossoma macropomum. Aquaculture. 2017; 469:79-87.; Pedrazzani, Neto, 2016Pedrazzani AS, Neto AO. The anaesthetic effect of camphor (Cinnamomum camphora), clove (Syzygium aromaticum) and mint (Mentha arvensis) essential oils on clown anemonefish, Amphiprion ocellaris (Cuvier 1830). Aquac Res . 2016; 47(3):769-76.; Cunha et al., 2017Cunha JA, Scheeren CA, Salbego J, Gressler LT, Madaloz LM, Bandeira-Junior G et al. Essential oils of Cunila galioides and Origanum majorana as anesthetics for Rhamdia quelen: efficacy and effects on ventilation and ionoregulation. Neotrop Ichthyol. 2017; 15(1):e160076. Available from: https://dx.doi.org/10.1590/1982-0224-20160076
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; Saccol et al., 2017Saccol EMH, Toni C, Pês T, Ourique GM, Gressler LT, Silva LVF et al. Anaesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils on tambaqui (Colossoma macropomum). Aquac Res . 2017; 48(5):2012-31.; Souza et al., 2017Souza CF, Baldissera MD, Salbego J, Lopes JM, Vaucher RA, Mourão RHV et al. Physiological responses of Rhamdia quelen (Siluriformes: Heptapteridae) to anesthesia with essential oils from two different chemotypes of Lippia alba. Neotrop Ichthyol . 2017; 15(1):e160083. Available from: https://dx.doi.org/10.1590/1982-0224-20160083.
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; Bodur et al., 2018Bodur T, Afonso JM, Montero D, Navarro A. Assessment of effective dose of new herbal anesthetics in two marine aquaculture species: Dicentrarchus labrax and Argyrosomus regius. Aquaculture . 2018; 482:78-82.), as well as some of their main compounds, as menthol (Façanha, Gomes, 2005Façanha MF, Gomes LC. A eficácia do mentol como anestésico para tambaqui (Colossoma macropomum, Characiformes: Characidae). Acta Amaz. 2005; 35(1):71-05.; Mazandarani, Hoseini, 2017Mazandarani M, Hoseini SM, Ghomshani MD. Effects of linalool on physiological responses of Cyprinus carpio (Linnaeus, 1758) and water physico-chemical parameters during transportation. Aquac Res . 2017; 48(12):5775-81.), globulol (Silva et al., 2013aSilva LL, Garlet QI, Benovit SC, Dolci G, Mallmann CA, Bürger ME et al. Sedative and anesthetic activities of the essential oils of Hyptis mutabilis (Rich.) Briq. and their isolated components in silver catfish (Rhamdia quelen). Braz J Med Biol Res . 2013a; 46(9):771-09.), (+)-spathulenol (Benovit et al., 2015Benovit SC, Silva LL, Salbego J, Loro VL, Mallmann CA, Baldisserotto B, Flores EM, Heinzmann BM. Anesthetic activity and bio-guided fractionation of the essential oil of Aloysia gratissima (Gillies & Hook.) Tronc. in silver catfish Rhamdia quelen. An Acad Bras Ciênc. 2015; 87(3):1675-89.), myrcene (Mirghaed et al., 2016Mirghaed AT, Ghelichpour M, Hoseini SM. Myrcene and linalool as new anesthetic and sedative agents in common carp, Cyprinus carpio - comparison with eugenol. Aquaculture . 2016; 464:165-70.), (+)-dehydrofukinone (Garlet et al., 2016Garlet QI, Pires LC, Silva DT, Spall S, Gressler LT, Bürger ME et al. Effect of (+)-dehydrofukinone on GABA(A) receptors and stress response in fish model. Braz J Med Biol Res . 2016; 49(1):e4872. Available from: http://dx.doi.org/10.1590/1414-431X20154872
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), thymol, carvacrol (Bianchini et al., 2017Bianchini AE, Garlet QI, Cunha JA, Bandeira-Junior G, Brusque ICM, Salbego J et al. Monoterpenoids (thymol, carvacrol and S-(+)-linalool) with anesthetic activity in silver catfish (Rhamdia quelen): evaluation of acetylcholinesterase and GABAergic activity. Braz J Med Biol Res. 2017; 50(12):e6346. Available from: http://dx.doi.org/10.1590/1414-431X20176346
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), and 1,8-cineole (Mazandarani, Hoseini, 2017Mazandarani M, Hoseini SM. Menthol and 1,8-cineole as new anaesthetics in common carp, Cyprinus carpio (Linnaeus, 1758). Aquac Res . 2017; 48(6):3041-51.).
Synthetic linalool (a mix of S-(+) and R-(-) isomers) presented anesthetic efficacy in common carp (Cyprinus carpio) (Mirghaed et al., 2016Mirghaed AT, Ghelichpour M, Hoseini SM. Myrcene and linalool as new anesthetic and sedative agents in common carp, Cyprinus carpio - comparison with eugenol. Aquaculture . 2016; 464:165-70.) and S-(+)-linalool isolated from the EO of Lippia alba had a similar sedation profile to the EO of this plant at a proportional linalool concentration in silver catfish (Rhamdia quelen) (Heldwein et al., 2014Heldwein CG, Silva LL, Gai EZ, Roman C, Parodi TV, Bürger ME et al. S-(+)-linalool from Lippia alba: sedative and anesthetic for silver catfish (Rhamdia quelen). Vet Anaesth Analg. 2014; 41(6):621-09.). The sedation was induced in silver catfish without differences between the compounds; however, R-(-)-linalool promoted faster anesthesia (Silva et al., 2017Silva LL, Balconi LS, Gressler LT, Garlet QI, Sutili FJ, Vargas AC et al. S-(+)- and R-(-)-linalool: a comparison of the in vitro anti-Aeromonas hydrophila activity and anesthetic properties in fish. An Acad Bras Cienc. 2017; 89(1):203-12.).
Experiments with rodents demonstrated the sedative effect of the EO of Aniba rosaeodora Ducke (EOAR, rosewood oil), a large tree native to the Amazon region (Almeida et al. 2009Almeida RN, Araujo DA, Goncalves JC, Montenegro FC, Sousa DP, Leite JR et al. Rosewood oil induces sedation and inhibits compound action potential in rodents. J Ethnopharmacol. 2009; 124(3):440-03.). The major component of EOAR is linalool (90.0%, with 80% being S-(+) and 20% being R-(-) isomers) (Lupe et al., 2008Lupe F, Souza R, Barata L. Seeking a sustainable alternative to Brazilian rosewood. Perfum Flavor. 2008; 33:40-03.; Fidelis et al., 2013Fidelis CHV, Sampaio PTB, Krainovic PM, Augusto F, Barata LES. Correlation between maturity of tree and GC×GC-qMS chemical profiles of essential oil from leaves of Aniba rosaeodora Ducke. Microchemical J. 2013; 109:73-07.). The EO of Aniba parviflora (Meisn.) Mez. (EOAP), another tree morphologically similar to A. rosaeodora (Galaverna et al., 2015Galaverna RS, Sampaio PTB, Barata LES, Eberlin MN, Fidelis CHV. Differentiation of two morphologically similar Amazonian Aniba species by mass spectrometry leaf fingerprinting. Anal Methods. 2015; 7(5):1984-90.), also contains linalool (29.6%) as main compound, as well as E-caryophyllene (10.9%), α-phellandrene (10.5%), β-selinene (8.0%), jinkoh eremol (7.2%) and p-cymene (6.3%) (Souza, 2010Souza RCZ. Avaliação das frações voláteis de espécies de Aniba por microextração em fase sólida acoplada a cromatografia gasosa (SPME-CG) e cromatografia gasosa bidimensional abrangente (CG × CG). [Master Thesis]. Campinas, SP: Instituto de Química - Universidade de Campinas; 2010.).
Tambaqui Colossoma macropomum (Cuvier, 1816) is the most raised native species in Brazil, mainly in the Amazon region (Valladão et al., 2016Valladão GMR, Gallani SU, Pilarski F. South American fish for continental aquaculture. Rev Aquacult. 2016, doi: 10.1111/raq.12164.
https://doi.org/10.1111/raq.12164...
), where both Aniba trees can be found. Tambaqui has been used as a model of Amazonian fish in studies of anesthesia with vegetable extractives and isolated compounds (Façanha, Gomes, 2005Façanha MF, Gomes LC. A eficácia do mentol como anestésico para tambaqui (Colossoma macropomum, Characiformes: Characidae). Acta Amaz. 2005; 35(1):71-05.; Roubach et al., 2005Roubach R, Gomes LC, Fonseca FAL, Val AL. Eugenol as an efficacious anaesthetic for tambaqui, Colossoma macropomum (Cuvier). Aquac Res . 2005; 36(11):1056-61.; Inoue et al., 2011Inoue LAKA, Boijink CL, Ribeiro PT, Silva AMD, Affonso EG. Avaliação de respostas metabólicas do tambaqui exposto ao eugenol em banhos anestésicos. Acta Amaz . 2011; 41(2):327-32.; Pádua et al., 2013Pádua SB, Dias Neto J, Sakabe R, Claudiano GS, Chagas EC, Pilarski F. Variáveis hematológicas em tambaquis anestesiados com óleo de cravo e benzocaína. Pesqui Agropec Bras. 2013; 48(8):1171-74.; Barbas et al., 2016Barbas LAL, Stringhetta GR, Garcia LO, Figueiredo MRC, Sampaio LA. Jambu, Spilanthes acmella as a novel anaesthetic for juvenile tambaqui, Colossoma macropomum: Secondary stress responses during recovery. Aquaculture . 2016; 456:70-05., 2017Barbas LAL, Maltez LC, Stringhetta GR, Garcia LO, Monserrat JM, Silva DT et al. Properties of two plant extractives as anaesthetics and antioxidants for juvenile tambaqui Colossoma macropomum. Aquaculture. 2017; 469:79-87.; Saccol et al., 2017Saccol EMH, Toni C, Pês T, Ourique GM, Gressler LT, Silva LVF et al. Anaesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils on tambaqui (Colossoma macropomum). Aquac Res . 2017; 48(5):2012-31.). Consequently, the aim of the present study is to verify the anesthetic efficacy of EOAR, EOAP and one of their main compounds, linalool, in two forms: synthetic and extracted from EOAR (linalool-AR).
Material and Methods
Essential oils and linalool. The EOAR and EOAP extraction from the leaves and their analysis was performed as described by Fidelis et al. (2013Fidelis CHV, Sampaio PTB, Krainovic PM, Augusto F, Barata LES. Correlation between maturity of tree and GC×GC-qMS chemical profiles of essential oil from leaves of Aniba rosaeodora Ducke. Microchemical J. 2013; 109:73-07.) and Souza (2010Souza RCZ. Avaliação das frações voláteis de espécies de Aniba por microextração em fase sólida acoplada a cromatografia gasosa (SPME-CG) e cromatografia gasosa bidimensional abrangente (CG × CG). [Master Thesis]. Campinas, SP: Instituto de Química - Universidade de Campinas; 2010.), respectively. The synthetic linalool [(±)-3,7-Dimethyl-1,6-octadien-3-ol, (±)-3,7-Dimethyl-3-hydroxy-1,6-octadiene] was purchased from Sigma-Aldrich. Linalool-AR was extracted from EOAR as described by Zellner et al. (2006Zellner BD, Presti LM, Barata LES, Dugo P, Dugo G, Mondello L. Evaluation of leaf-derived extracts as an environmentally sustainable source of essential oils by using gas chromatography−mass spectrometry and enantioselective gas chromatography−olfactometry. Anal Chem. 2006; 78(3):883-90.).
Fish and culture conditions. The experiments were conducted in a fish farm (Santarém, Pará, Brazil) under authorization of Secretaria Estadual de Desenvolvimento Agropecuário e de Pesca (SEDAP, Pará, Brazil) and Instituto Chico Mendes de Conservação da Biodiversidade (ICMBio)/Sistema de Autorização e Informação em Biodiversidade (SISBIO, nº 35427-1). Capture and maintenance of juveniles of tambaqui (voucher number UFOPA-I 000143) was performed as described by Saccol et al. (2017Saccol EMH, Toni C, Pês T, Ourique GM, Gressler LT, Silva LVF et al. Anaesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils on tambaqui (Colossoma macropomum). Aquac Res . 2017; 48(5):2012-31.). The experimental protocol was approved by the Committee on Ethics in Research of Universidade do Estado do Pará (Campus XII, Santarém) under registration nº 42/2012. All animal manipulations were carried out in compliance with the guidelines of ethics committee mentioned above.
Anesthesia induction and recovery. Tambaqui juveniles (2.0 ± 0.2 g, 5.0 ± 0.2 cm) were individually transferred to continuously aerated aquaria containing 500 mL of water and 25, 50, 100 or 200 µL L-1 of EOAR or 50, 100, 200 or 300 µL L-1 of EOAP or synthetic linalool or linalool-AR, first diluted in ethanol (1:10) to enable better dilution in water. Previous tests were made with 200 µL L-1 of both EOs and compounds, based on studies of Cunha et al. (2010Cunha MA, Barros FMC, Garcia LO, Veeck APL, Heinzmann BM, Loro VL et al. Essential oil of Lippia alba: A new anesthetic for silver catfish, Rhamdia quelen. Aquaculture . 2010; 306:403-6.) with the EO of Lippia alba chemotype linalool (around 47% linalool) and Heldwein et al. (2014Heldwein CG, Silva LL, Gai EZ, Roman C, Parodi TV, Bürger ME et al. S-(+)-linalool from Lippia alba: sedative and anesthetic for silver catfish (Rhamdia quelen). Vet Anaesth Analg. 2014; 41(6):621-09.) and Silva et al. (2017Silva LL, Balconi LS, Gressler LT, Garlet QI, Sutili FJ, Vargas AC et al. S-(+)- and R-(-)-linalool: a comparison of the in vitro anti-Aeromonas hydrophila activity and anesthetic properties in fish. An Acad Bras Cienc. 2017; 89(1):203-12.) with linalool in silver catfish. Based on the time to reach deep anesthesia using this concentration, the concentration range to be tested with each EO or compound was chosen. The maximum evaluation time was 30 min, and 10 juveniles were used for each concentration tested; each juvenile was only used once. Ethanol alone did not produce any anesthetic effect in tambaqui (Saccol et al., 2017Saccol EMH, Toni C, Pês T, Ourique GM, Gressler LT, Silva LVF et al. Anaesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils on tambaqui (Colossoma macropomum). Aquac Res . 2017; 48(5):2012-31.). The evaluation of each anesthesia stage was an adaptation of Small (2003Small BC. Anesthetic efficacy of metomidate and comparison of plasma cortisol responses to tricaine methanesulfonate, quinaldine and clove oil anesthetized channel catfish Ictalurus punctatus. Aquaculture . 2003; 218(1-4):177-85.): stage I - light sedation, with decreased reactivity to external stimuli; stage II - deep sedation, with partial loss of equilibrium and erratic swimming; and stage III - deep anesthesia, with a total loss of equilibrium, cessation of locomotion and no response to tactile stimuli (pressure of a glass rod on the caudal peduncle).
Long-term exposure. The long-term exposure was performed to analyze the use of the EOs and linalool for fish transport. Juveniles were maintained individually for 12 h (n=10 each concentration tested) in continuously aerated 1 L aquaria containing 5 or 10 µL L-1 of each tested EO or linalool. Tambaquis exposed to 25 µL L-1 EOAR reached deep sedation within 30 min (see results) and consequently the concentrations chosen for log-term exposure were lower than this concentration to avoid fish reaching anesthesia as time went by, which is not recommended for transportation (Becker et al., 2012Becker AG, Parodi TV, Heldwein CG, Zeppenfeld CC, Heinzmann BM, Baldisserotto B. Transportation of silver catfish, Rhamdia quelen, in water with eugenol and the essential oil of Lippia alba. Fish Physiol Biochem. 2012; 38(3):789-96.). Swimming behavior was observed every hour up to the end of the experiment, to check if fish showed agitation (sudden swimming bursts) or any loss of equilibrium. The temperature was 25.6-26.4oC and dissolved oxygen levels (YSI multiparameter- Professional plus-) were above 6.0 mg L-1 throughout the experiment.
Statistical analyses. The data are expressed as mean ± SEM. The evaluation of anesthetic activity was performed by regression analysis (concentration x time of anesthesia induction; concentration x time of recovery from anesthesia) using Sigma Plot 11.0 software. When no relationship was found, homogeneity of variances between concentrations was tested with the Levene test. Data were log transformed to obtain homoscedasticity and then were analyzed using one-way ANOVA and Tukey’s test. Comparisons between synthetic linalool and linalool-AR were tested by two-way ANOVA (linalool type x concentration). All statistical analyses were performed using the software Statistica 7.0. Minimum significance level was P<0.05.
Results
Anesthesia induction and recovery. No fish died during the course of the anesthesia experiment. Induction time for all anesthesia stages decreased as the concentrations of EOAR, EOAP, synthetic and linalool-AR were raised, but a significant relationship for concentration x anesthesia induction time was not observed in the stage of light sedation for EOAR and linalool-AR (Figs. 1-2). Fish exposed to 25 µL L-1 EOAR and 50 µL L-1 of EOAP and both linalools did not reach deep anesthesia (Tabs 1-2). Linalool-AR showed a longer time for anesthesia induction in some stages and for recovery at 100 and 200 µL L-1 (Tab. 2) in comparison to the synthetic mixture. Recovery time was lengthened as the concentration of EOAR and synthetic linalool increased and from 50 to 100 µL L-1 linalool-AR. The recovery time was not affected significantly by EOAP concentration (Tabs. 1-2). A significant relationship for concentration x recovery time was observed only in fish anesthetized with synthetic linalool (Fig. 2a).
Long-term exposure. Fish exposed to both concentrations of EOs and linalools did not show any loss of equilibrium or sudden swimming bursts throughout the 12 h of exposure.
Relationships of concentration x anesthesia induction or recovery time in tambaqui, Colossoma macropomum, exposed to the essential oils. a. Aniba rosaeodora (EOAR); deep sedation: y=24.8+(8300/x), r2=0.705, deep anesthesia: y=-102.3+(36527/x), r2=0.913. Light sedation and recovery: no significant relationship. b. Aniba parviflora (EOAP); light sedation: y=2.08+(6563/x), r2=0.832, deep sedation: y=13.2+(10098/x), r2=0.703, deep anesthesia: y=-79.8+(41067/x), r2=0.906. Recovery: no significant relationship. y = time to reach stage or recovery (s) and x = concentration (µL L-1).
Relationships concentration x anesthesia induction or recovery time in tambaqui, Colossoma macropomum, exposed to the linalools. a. synthetic linalool; light sedation: y=4.4+(4281/x), r2=0.716, deep sedation: y=-22.5+(12252/x), r2=0.773, deep anesthesia: y=15.3+(17878/x), r2=0.669, recovery: y=43.9+0.66x+0.0015x2, r2=0.712. b. linalool extracted from Aniba rosaeodora; deep sedation: y=29.0+(6010/x), r2=0.784, deep anesthesia: y=-151.3+(60541/x), r2=0.873. Light sedation and recovery: no significant relationship. y = time to reach stage or recovery (s) and x = concentration (µL L-1).
Time required for induction and recovery from anesthesia using the essential oils of Aniba rosaeodora (EOAR) and Aniba parviflora (EOAP). Different letters in the column indicate significant difference between concentrations. One-way ANOVA and Tukey’s test (P<0.05). Stages in which there was a significant relationship concentration x time of anesthesia induction, no statistical comparison between concentrations was made.
Time required for induction and recovery from anesthesia using synthetic linalool and linalool extracted from the essential oil of Aniba rosaeodora (linalool-AR). *significantly different from synthetic linalool. Different letters in the column indicate significant difference between concentrations. One-way ANOVA and Tukey’s test (P<0.05). Stages in which there was a significant relationship concentration x time of anesthesia induction, no statistical comparison between concentrations was made.
Discussion
Time for tambaqui to reach anesthesia reduced as the concentration of EOAR, EOAP and both linalools increased. The same pattern was observed for juveniles of this species with the same size of the present study when exposed to the essential oils from the leaves of Myrcia sylvatica, Curcuma longa (Saccol et al., 2017Saccol EMH, Toni C, Pês T, Ourique GM, Gressler LT, Silva LVF et al. Anaesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils on tambaqui (Colossoma macropomum). Aquac Res . 2017; 48(5):2012-31.) and Nectandra grandiflora (Barbas et al., 2017Barbas LAL, Maltez LC, Stringhetta GR, Garcia LO, Monserrat JM, Silva DT et al. Properties of two plant extractives as anaesthetics and antioxidants for juvenile tambaqui Colossoma macropomum. Aquaculture. 2017; 469:79-87.), as well as 56-88 g specimens with the compounds eugenol (Roubach et al., 2005Roubach R, Gomes LC, Fonseca FAL, Val AL. Eugenol as an efficacious anaesthetic for tambaqui, Colossoma macropomum (Cuvier). Aquac Res . 2005; 36(11):1056-61.) and menthol (Façanha, Gomes, 2005Façanha MF, Gomes LC. A eficácia do mentol como anestésico para tambaqui (Colossoma macropomum, Characiformes: Characidae). Acta Amaz. 2005; 35(1):71-05.). The increased concentration of synthetic linalool and the S-(+) and R-(-) isomers also decreased anesthesia induction time in common carp (4-5 g) (Mirghaed et al., 2016Mirghaed AT, Ghelichpour M, Hoseini SM. Myrcene and linalool as new anesthetic and sedative agents in common carp, Cyprinus carpio - comparison with eugenol. Aquaculture . 2016; 464:165-70.) and silver catfish (4-9 g) (Heldwein et al., 2014Heldwein CG, Silva LL, Gai EZ, Roman C, Parodi TV, Bürger ME et al. S-(+)-linalool from Lippia alba: sedative and anesthetic for silver catfish (Rhamdia quelen). Vet Anaesth Analg. 2014; 41(6):621-09.; Silva et al., 2017Silva LL, Balconi LS, Gressler LT, Garlet QI, Sutili FJ, Vargas AC et al. S-(+)- and R-(-)-linalool: a comparison of the in vitro anti-Aeromonas hydrophila activity and anesthetic properties in fish. An Acad Bras Cienc. 2017; 89(1):203-12.), respectively. The increase in time for anesthesia recovery with the increase in EOAR and synthetic linalool concentrations in tambaqui was also observed in this species when anesthetized with the essential oil of N. grandiflora (Barbas et al., 2017Barbas LAL, Maltez LC, Stringhetta GR, Garcia LO, Monserrat JM, Silva DT et al. Properties of two plant extractives as anaesthetics and antioxidants for juvenile tambaqui Colossoma macropomum. Aquaculture. 2017; 469:79-87.), eugenol (Roubach et al., 2005Roubach R, Gomes LC, Fonseca FAL, Val AL. Eugenol as an efficacious anaesthetic for tambaqui, Colossoma macropomum (Cuvier). Aquac Res . 2005; 36(11):1056-61.) and menthol (Façanha, Gomes, 2005Façanha MF, Gomes LC. A eficácia do mentol como anestésico para tambaqui (Colossoma macropomum, Characiformes: Characidae). Acta Amaz. 2005; 35(1):71-05.) and in silver catfish anesthetized with both linalool isomers (Silva et al., 2017Silva LL, Garlet QI, Benovit SC, Dolci G, Mallmann CA, Bürger ME et al. Sedative and anesthetic activities of the essential oils of Hyptis mutabilis (Rich.) Briq. and their isolated components in silver catfish (Rhamdia quelen). Braz J Med Biol Res . 2013a; 46(9):771-09.). However, recovery time showed no relationship with synthetic linalool concentration in common carp (Mirghaed et al., 2016Mirghaed AT, Ghelichpour M, Hoseini SM. Myrcene and linalool as new anesthetic and sedative agents in common carp, Cyprinus carpio - comparison with eugenol. Aquaculture . 2016; 464:165-70.) with either concentration of the essential oils of M. sylvatica and C. longa in tambaqui (Saccol et al., 2017Saccol EMH, Toni C, Pês T, Ourique GM, Gressler LT, Silva LVF et al. Anaesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils on tambaqui (Colossoma macropomum). Aquac Res . 2017; 48(5):2012-31.). Recovery time from anesthesia varies with the anesthetic and the species analyzed; the present study demonstrated that the EOAP concentration did not change recovery time in tambaqui.
Exposure to 25 and 50 µL L-1 EOAR was sufficient to induce sedation and deep anesthesia in tambaqui; therefore, this essential oil was more efficient than EOAP, synthetic linalool and linalool-AR because they need two-fold concentrations to provoke the same effects. Apparently, some minor compounds of EOAR (all less than 10%) contribute to improve the anesthetic efficacy of linalool. EOAP anesthetic induction time is within the synthetic linalool and linalool-AR. As EOAP contains only 29.6% linalool, the other compounds are probably involved in its anesthetic effect. E-caryophyllene is a major compound of the essential oil of L. alba linalool chemotype, which anesthetizes silver catfish (Souza et al., 2017Souza CF, Baldissera MD, Salbego J, Lopes JM, Vaucher RA, Mourão RHV et al. Physiological responses of Rhamdia quelen (Siluriformes: Heptapteridae) to anesthesia with essential oils from two different chemotypes of Lippia alba. Neotrop Ichthyol . 2017; 15(1):e160083. Available from: https://dx.doi.org/10.1590/1982-0224-20160083.
https://dx.doi.org/10.1590/1982-0224-201...
), and p-cymene is an important compound of the essential oil of Cinnamomum camphora, which has anesthetic activity in clown anemonefish (Amphiprion ocellaris) (Pedrazzani, Neto, 2016Pedrazzani AS, Neto AO. The anaesthetic effect of camphor (Cinnamomum camphora), clove (Syzygium aromaticum) and mint (Mentha arvensis) essential oils on clown anemonefish, Amphiprion ocellaris (Cuvier 1830). Aquac Res . 2016; 47(3):769-76.). α-phellandrene and p-cymene are major compounds of the essential oil of Foeniculum vulgare, which reduces anxiety in mice (Mesfin et al., 2014Mesfin M, Asres K, Shibeshi W. Evaluation of anxiolytic activity of the essential oil of the aerial part of Foeniculum vulgare Miller in mice. BMC Complement Altern Med. 2014; 14:310. Available from: https://doi.org/10.1186/1472-6882-14-310
https://doi.org/10.1186/1472-6882-14-310...
), β-selinene is a major compound of the essential oil of M. sylvatica with anesthetic properties (Saccol et al., 2017Saccol EMH, Toni C, Pês T, Ourique GM, Gressler LT, Silva LVF et al. Anaesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils on tambaqui (Colossoma macropomum). Aquac Res . 2017; 48(5):2012-31.). Peritoneal and intracerebroventricular injection of jinkoh eremol decreased both methamphetamine- and apomorphine-induced spontaneous motility in mice (Okugawa et al., 1996Okugawa H, Ueda R, Matsumoto K, Kawanishi K, Kato A. Effect of jinkoh-eremol and agarospirol from agarwood on the central nervous system in mice. Planta Med. 1996; 62(1):2-6.).
The essential oil of M. sylvatica needed the same concentrations as EOAP to induce sedation and deep anesthesia in tambaqui (Saccol et al., 2017Saccol EMH, Toni C, Pês T, Ourique GM, Gressler LT, Silva LVF et al. Anaesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils on tambaqui (Colossoma macropomum). Aquac Res . 2017; 48(5):2012-31.), but the essential oils of C. longa and N. grandiflora were less efficient in specimens of the same size (Saccol et al., 2017Saccol EMH, Toni C, Pês T, Ourique GM, Gressler LT, Silva LVF et al. Anaesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils on tambaqui (Colossoma macropomum). Aquac Res . 2017; 48(5):2012-31.; Barbas et al., 2017Barbas LAL, Maltez LC, Stringhetta GR, Garcia LO, Monserrat JM, Silva DT et al. Properties of two plant extractives as anaesthetics and antioxidants for juvenile tambaqui Colossoma macropomum. Aquaculture. 2017; 469:79-87.). The waxy extract of the flowers of Spilanthes acmella presented a better anesthetic efficiency in 46 g tambaqui because 5 mg L-1 was sufficient to induce sedation and deep anesthesia (Barbas et al., 2016Barbas LAL, Stringhetta GR, Garcia LO, Figueiredo MRC, Sampaio LA. Jambu, Spilanthes acmella as a novel anaesthetic for juvenile tambaqui, Colossoma macropomum: Secondary stress responses during recovery. Aquaculture . 2016; 456:70-05., 2017Barbas LAL, Maltez LC, Stringhetta GR, Garcia LO, Monserrat JM, Silva DT et al. Properties of two plant extractives as anaesthetics and antioxidants for juvenile tambaqui Colossoma macropomum. Aquaculture. 2017; 469:79-87.).
Exposure to 50 µL L-1 synthetic linalool also induced light sedation in common carp, but only fish exposed to 200 µL L-1 reached deep anesthesia (Mirghaed et al., 2016Mirghaed AT, Ghelichpour M, Hoseini SM. Myrcene and linalool as new anesthetic and sedative agents in common carp, Cyprinus carpio - comparison with eugenol. Aquaculture . 2016; 464:165-70.), and in both situations induction time was longer than for tambaqui. Time to induce light sedation with 60 µL L-1 S-(+) and R-(-)-linalool in silver catfish (Silva et al., 2017Silva LL, Balconi LS, Gressler LT, Garlet QI, Sutili FJ, Vargas AC et al. S-(+)- and R-(-)-linalool: a comparison of the in vitro anti-Aeromonas hydrophila activity and anesthetic properties in fish. An Acad Bras Cienc. 2017; 89(1):203-12.) is similar to that for synthetic and linalool-AR in tambaqui, but silver catfish took longer to reach deep anesthesia with both linalool isomers. However, in this context, it is noteworthy that tambaqui experiments were carried out at higher water temperatures than those used in silver catfish, which can explain the differences observed in the recovery time. The fish size and water parameters from these studies were also different, and as anesthesia induction time may be affected by these parameters (Gomes et al., 2011Gomes DP, Chaves BW, Becker AG, Baldisserotto B. Water parameters affect anaesthesia induced by eugenol in silver catfish, Rhamdia quelen. Aquac Res. 2011; 42(6):878-86.), they may also explain these differences.
The R-(-)-linalool promoted faster anesthesia in silver catfish within the 180-500 µL L-1 range compared to the S-(+) isomer (Silva et al., 2017Silva LL, Balconi LS, Gressler LT, Garlet QI, Sutili FJ, Vargas AC et al. S-(+)- and R-(-)-linalool: a comparison of the in vitro anti-Aeromonas hydrophila activity and anesthetic properties in fish. An Acad Bras Cienc. 2017; 89(1):203-12.). The same authors also demonstrated that synthetic linalool contains almost equal concentrations of both isomers. Thus, the higher percentage of the R-(-)-linalool isomer in the synthetic sample explains why linalool-AR (20% of the R-(-)-isomer) took slightly longer to induce some stages of anesthesia in tambaqui.
The transport of common carp for 3 h with 50-200 µL L-1 synthetic linalool was not recommended because it decreased dissolved oxygen levels, has no benefits in preventing ion loss and increased stress compared to the control fish (Mazandarani et al., 2017Mazandarani M, Hoseini SM, Ghomshani MD. Effects of linalool on physiological responses of Cyprinus carpio (Linnaeus, 1758) and water physico-chemical parameters during transportation. Aquac Res . 2017; 48(12):5775-81.). However, exposure of tambaqui for 12 h to 5 or 10 µL L-1 of EOAR, EOAP, synthetic linalool and linalool-AR did not change equilibrium and swimming behavior. Therefore, these concentrations may be tested in the transport of this species evaluating ionoregulatory parameters and stress response.
In conclusion, considering anesthesia induction and recovery time, EOAR, EOAP, synthetic linalool and linalool-AR can be used as anesthetics and probably as sedatives for the transport in tambaqui. Additional studies must investigate whether these anesthetics can avoid or reduce stress provoked by handling and/or transportation.
Acknowledgments
We thank INCT-ADAPTA 2 (Conselho Nacional de Desenvolvimento Científico e Tecnológico/Fundação de Amparo à Pesquisa no Estado do Amazonas). B. Baldisserotto received CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico) research fellowship and L.L. Silva and C. Toni CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) PhD fellowships.
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Publication Dates
-
Publication in this collection
2018
History
-
Received
16 Oct 2017 -
Accepted
23 Feb 2018