Behavioral, electrocardiographic evaluation, and mechanism of action of Curcuma longa essential oil in juvenile tambaqui Colossoma macropomum (Cuvier, 1818) subjected to immersion baths at different concentrations

Behavioral, electrocardiographic evaluation, and mechanism of action of Curcuma longa essential oil in juvenile tambaqui Colossoma macropomum (Cuvier, 1818) subjected to immersion baths at different concentrations | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Behavioral, electrocardiographic evaluation, and mechanism of action of Curcuma longa essential oil in juvenile tambaqui Colossoma macropomum (Cuvier, 1818) subjected to immersion baths at different concentrations Luciana Esquerdo Cerqueira, Daniella Bastos de Araújo, Luciana Eiró-Quirino, and 10 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5154092/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The use of natural agents as anesthetics in fish management aims to minimize stress during procedures. This study aims to investigate the behavioral, electrocardiographic characteristics, and the pharmacological mechanism of action of Curcuma longa essential oil (CLEO) in Colossoma macropomum . The study was conducted on juvenile Colossoma macropomum (18.13 ± 2.1g) (n = 153), using CLEO concentrations of 125 µL.L⁻¹, 150 µL.L⁻¹, 175 µL.L⁻¹, 200 µL.L⁻¹, and 250 µL.L⁻¹, analyzing anesthetic induction and recovery behavior (Experiment I), electrocardiogram (Experiment II), and the underlying mechanism of action (Experiment III). Fish exposed to CLEO concentrations reached a deep anesthesia stage in a concentration-dependent manner. However, the effects obtained during anesthetic induction were reversible, both in behavioral parameters and cardiac activity. The mechanism of action was observed after administration of flumazenil 1 mg/kg i.p., which showed greater resistance to loss of postural reflex and shorter latency for recovery. The mechanism of action indicates involvement of inhibitory GABA neurotransmission. The cardiac effects at the concentrations used were compatible with anesthesia, but no arrhythmias occurred that could compromise the hemodynamics of the fish, demonstrating its safety for short-duration anesthesia induction in Colossoma macropomum . Curcuma longa electrocardiographic behavioral Colossoma macropomum tambaqui Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 1 Introduction Aquaculture activities subject fish to various potential stressors. Handling and transport procedures, for instance, can result in injuries that serve as entry points for infections and bacteria (Ribeiro et al., 2016 ; Barbas et al., 2017 ; Primadona et al., 2017 ). Additionally, high stocking densities in aquaculture environments increase stress levels in fish (Zahl et al., 2012 ), which significantly impacts their physiology and survival (Harmon, 2009 ). Stress contributes to elevated fish mortality rates (Delbon & Paiva, 2012 ; Abdel-Aziz, 2013 ), potentially leading to significant resource and productivity losses in the production chain (Davis, 2010 ; Becker et al., 2012 ; Hohlenwerger et al., 2017 ; Teixeira et al., 2018 ). As such, anesthetic strategies have become crucial in ensuring the success of aquaculture by promoting animal welfare during and after handling procedures (Bianchini et al., 2017 ; Hoseini et al., 2019 ). Among anesthetics used in fish, those of natural origin have been widely studied due to their efficacy in transport when administered in low doses to induce anesthesia (Suwandi, Nugraha, & Zulfamy, 2013 ; Aydın et al., 2015 ). Additionally, they increase post-administration survival rates (Hasan et al., 2016), exhibit low tissue residue in animals, and reduce the number of pollutants released into the environment (Ramanayaka & Atapattu, 2006 ; Rezende, Pascoal, Vianna, & Lanna, 2017 ). One such natural product is Curcuma longa essential oil (CLEO), which has demonstrated potential for use in fish anesthesia. According to Schramm et al. ( 2013 ), certain constituents of this species may exert pharmacological effects possibly associated with the GABAergic system. Curcuma longa is well-known for its diverse pharmacological and therapeutic properties (Aggarwal et al., 2009), including antioxidant, anticancer, antiatherosclerotic, antidiabetic, anti-inflammatory, immunomodulatory, and sedative activities (Yu, Kong & Chen, 2002 ; Vijayasteltar et al., 2011 ; Priya et al., 2012 ; Parveen et al., 2013 ). Tambaqui ( Colossoma macropomum ) is the most widely farmed native fish species in Brazil and across several nations in South and Central America, according to reports from FAO ( 2014 ) and IBGE ( 2020 ). Belonging to the family Serrasalmidae, tambaqui is endemic to the Amazon and Orinoco River basins (Merola, 1987 ; Goulding, 1982 ). This Amazonian species exhibits high growth rates and feed conversion efficiency, as demonstrated by Merola and Cantelmo ( 1987 ). The growing interest in tambaqui aquaculture can be attributed to its high meat quality, growth rate, and feed efficiency (Guimarães et al., 2014 ). Furthermore, studies by Barbas et al. ( 2016 a, 2016 b) suggest that tambaqui is a suitable model for anesthetic studies in tropical freshwater species. According to data from the Brazilian Aquaculture Association (2024), Brazil produced approximately 887,029 tons of native fish in 2023, with tambaqui being the second most exported species, demonstrating an 809% increase in production (PEIXE BR, 2024 ) compared to the previous year. This growing profitability has spurred interest in developing more efficient farming strategies to enhance productivity, as well as the health and welfare of these fish (Guilherme et al., 2022 ; Luz et al., 2021 ; Nakayama et al., 2022 ; Vieira et al., 2022 ). The concern for fish welfare is highly relevant in anesthesia studies. Electrocardiography (ECG) is one of the tools used to ensure that vital functions are not compromised during anesthesia. Studies by Arnaout et al. ( 2007 ) and Koopman et al. ( 2021 ) noted the similarity between the ECG waveforms (P waves, QRS complexes, and T waves) of adult zebrafish and humans. Additionally, zebrafish studies (Genge et al., 2016 ; Le et al., 2022 ) have been successful in screening drug-induced cardiac damage, including drug-induced QT interval prolongation. Thus, this study evaluates the behavioral and electrocardiographic responses of tambaqui ( Colossoma macropomum ) subjected to immersion baths with varying concentrations of Curcuma longa essential oil, aiming to ensure the safe use of this oil in anesthetic induction and recovery. 2 Materials and methods 2.1 Experimental Animals The experimental subjects used in this study were Colossoma macropomum (tambaqui) (n = 108, males and females). The fish were housed in aquaria at the Experimental Facility of the Laboratory of Pharmacology and Toxicology of Natural Products at the Federal University of Pará (ICB/UFPA) under controlled temperature conditions (25–27°C) and a 12-hour light/12-hour dark photoperiod. The animals were fed commercial feed (32% protein) twice daily to satiation. Unconsumed feed and feces were removed by siphoning, and approximately 30% of the tank water was renewed daily with filtered water from the same source. During a 10-day acclimatization period, water quality parameters were monitored, including water temperature (26.3°C) and pH (7.6). The research project was submitted to and approved by the UFPA Research Ethics Committee file number 9261280624 [ID 002674]. 2.2 Organoleptic Properties and Chromatographic Analysis Curcuma longa essential oil (CLEO) was purchased from Laszlo Aromatherapy Laboratory (Brazil, CNPJ: 07.997.093/0001–10). The oil was extracted by steam distillation, and chromatographic analysis was performed using Gas Chromatography-Mass Spectrometry (GC-MS) on an AGILENT 7820A GC system under the following conditions: HP-5 column (30 m x 0.32 mm x 0.25 µm; AGILENT), column temperature: 70°C (0 min), 3°C/min up to 250°C, injector temperature: 250°C, split ratio: 1:50, detector temperature: FID 260°C, injection volume: 1 µL (1% in chloroform). The phytoconstituents of the oil are listed in Table 1, with the major component being ar-turmerone (Fig. 1 ). Table I Composition of Curcuma longa essential oil (CLEO) Retention time order Identification % 1 Careno 0.4 2 1,8-Cineol 0.4 3 Farneseno 7.2 4 α-curcumeno 5.7 5 β-curcumeno 13.0 6 α-bergamoteno 2.6 7 β-bisaboleno 2.7 8 Sesquifelandreno 3.2 9 Curlona 8.7 10 ar-turmerona 32.5 11 Turmerol 1.5 12 Atlantona 1.0 2.3 Experimental Design 2.3.1 Experiment with Curcuma longa Essential Oil (CLEO) Juvenile tambaqui ( Colossoma macropomum ) weighing 18.13 ± 2.1 g were randomly distributed into the following treatments: a) Control; b) Vehicle group (fish subjected to an immersion bath in 3 mL of 70% alcohol diluted in 1 liter of aquarium water); c) Fish treated with CLEO at concentrations of 125 µL.L⁻¹, 150 µL.L⁻¹, 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹. All fish were subjected to anesthetic induction for a duration of 5 minutes, followed by a 5-minute recovery period in water without CLEO. Each treatment group consisted of 9 fish, for a total of 108 juveniles of Colossoma macropomum . 2.3.2 Experiment 1 - Behavioral Analysis of Anesthetic Induction and Recovery The latency for the loss of posture reflex, characterized by lateral decubitus, was evaluated in fish exposed to CLEO concentrations of 125 µL.L⁻¹, 150 µL.L⁻¹, 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹ (n = 9 per treatment). After anesthetic induction, the latency for recovery of the posture reflex was assessed. 2.3.3 Experiment 2 - Electrocardiogram (ECG) Analysis Cardiac function was monitored in seven groups: a) Control group; b) Vehicle group; c) Groups treated with CLEO at concentrations of 125 µL.L⁻¹, 150 µL.L⁻¹, 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹, totaling 63 animals. Custom electrodes made from 0.3 mm diameter 925 silver with a length of 10 mm were used. The negative reference electrode was positioned at the base of the heart, and the positive recording electrode at the heart apex. The reference electrode was attached to the ventral portion of the left opercular opening (0.2 mm from the opercular cavity), and the recording electrode was inserted 2.0 mm from the right opercular opening. Electrodes were connected to a high-impedance amplifier (Grass Technologies, Model P511), enabling analysis of heart rate (bpm), QRS complex amplitude (mV), QRS complex duration (ms), R-R intervals (ms), P-Q intervals (ms), and Q-T intervals (ms). 2.3.4 Recording and Analysis of ECG Data Electrodes were connected to a digital data acquisition system through a differential high-input impedance amplifier (Grass Technologies, Model P511), filtered at 0.3–300 Hz, amplified 2000x, and monitored with an oscilloscope (ProteK, Model 6510). Signals were digitized at a rate of 1 kHz and stored on a hard drive for later processing using specialized software (LabVIEW Express). Data analysis was performed with a custom Python program utilizing Numpy and Scipy libraries for mathematical processing, and Matplotlib for graph generation. The graphical interface was developed using the PyQt4 library (Hamoy et al., 2023 ). 2.3.5 Experiment 3 - Evaluation of Mechanism Underlying CLEO Effects To assess the GABAergic allosteric activity of CLEO, fish were pretreated with flumazenil (0.1 mg/mL, 1 mg/kg i.p.) 15 minutes prior to CLEO treatment at concentrations of 125 µL.L⁻¹, 150 µL.L⁻¹, 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹. Anesthetic induction was observed until the loss of the posture reflex, followed by recovery time assessment. 2.4 Statistical Analysis Data normality and homogeneity of variances were verified using the Kolmogorov-Smirnov and Levene tests, respectively. One-way ANOVA followed by Tukey's post-hoc test was performed to compare means. Statistical analysis was conducted using GraphPad Prism® 8, with significance levels set at *p < 0.05, **p < 0.01, and ***p < 0.001 for all comparisons. 3 Results The behavioral analysis revealed that CLEO induced a loss of posture reflex in fish, with higher doses resulting in shorter latencies for the onset of this reflex loss. Fish treated with 125 µL.L⁻¹ of CLEO exhibited a mean latency for loss of posture reflex of 169.0 ± 12.00 seconds, which was longer compared to the other groups. The group treated with 150 µL.L⁻¹ had a mean latency of 147.6 ± 11.90 seconds, while groups treated with 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹ showed significantly shorter latencies of 129.1 ± 11.99 seconds, 101.2 ± 7.48 seconds, and 90.56 ± 10.25 seconds, respectively (Fig. 2 A). Recovery of the posture reflex in the group treated with 125 µL.L⁻¹ of CLEO occurred in 123.9 ± 13.30 seconds, which was shorter than the recovery times in the other groups: 150 µL.L⁻¹ (168.9 ± 11.67 seconds), 175 µL.L⁻¹ (206 ± 14.27 seconds), 200 µL.L⁻¹ (245 ± 13.04 seconds), and 225 µL.L⁻¹ (275.4 ± 16.98 seconds). All groups showed concentration-dependent recovery times, with higher concentrations resulting in longer recovery periods. This indicates that the anesthetic effect was reversible but occurred more slowly in fish exposed to higher concentrations (Fig. 2 B). The normal electrocardiogram of tambaqui exhibited a sinus rhythm, with a mean heart rate of 95.22 ± 7.775 bpm. The P wave, QRS complex, and T wave were clearly identifiable (Fig. 3 A, B, and C). The R-R intervals (ms), P-Q intervals (ms), and Q-T intervals (ms) remained consistent with normal cardiac parameters, allowing for the assessment of treatment interference at increasing doses of CLEO. In the control group, the cardiac rhythm was sinusoidal throughout the 5-minute recording (Fig. 3 ). Cardiac activity in the control group exhibited a mean heart rate of 95.22 ± 7.77 bpm, which was similar to the vehicle group (p = 0.999), both maintaining a sinus rhythm with all cardiac deflections visible on the electrocardiogram (Fig. 3 A, B, C, and 4 A). The 30-second amplification allowed observation of all ECG elements: the P wave representing atrial contraction, the QRS complex indicating ventricular contraction, and the T wave representing ventricular repolarization. The 5-second amplification facilitated the evaluation of intervals during immersion treatment with CLEO and subsequent recovery (Fig. 4 A, B, C, D, E, and F). During immersion treatment with 125 µL.L⁻¹, fish exhibited a 15.28% decrease in heart rate compared to the control group (Fig. 4 B). The group treated with 150 µL.L⁻¹ of CLEO showed a 24.15% reduction (Fig. 4 C), while the group treated with 175 µL.L⁻¹ exhibited a 24.61% decrease (Fig. 4 D). A 36.28% reduction was observed in the group treated with 200 µL.L⁻¹ (Fig. 4 E), and the group treated with 225 µL.L⁻¹ presented a 45.85% decrease (Fig. 4 F). Fish treated with CLEO displayed bradycardia while maintaining a sinus rhythm, with the intensity of bradycardia increasing alongside the concentration (Figs. 4 B, C, D, E, and F). The cardiac frequency decreased during treatment with increasing concentrations of CLEO. The control group had a mean frequency of 95.22 ± 7.775 bpm, which was similar to the vehicle group (p = 0.999), but higher than the treated groups. The group treated with 125 µL.L⁻¹ had a mean frequency of 80.67 ± 3.0 bpm, significantly higher than the other treated groups. The 150 µL.L⁻¹ group (72.22 ± 2.72 bpm) was similar to the 175 µL.L⁻¹ group (p = 0.999) but higher than the groups treated with 200 µL.L⁻¹ and 225 µL.L⁻¹. The 225 µL.L⁻¹ group had the lowest frequency compared to all other groups (Fig. 5 A). The mean amplitude of the QRS complex in the control group was 1.634 ± 0.23 mV, which was similar to the other groups (F(6, 56) = 2.348; p = 0.0430) (Fig. 5 B). The mean RR interval for the control group was 633.1 ± 47.59 ms, similar to the vehicle group (p = 0.999), but lower than the other treated groups. The group treated with 125 µL.L⁻¹ had a mean RR interval of 743.9 ± 28.50 ms, which was lower than the other treated groups. The 150 µL.L⁻¹ group (831.4 ± 30.49 ms) was similar to the 175 µL.L⁻¹ group (p = 0.999), while the 225 µL.L⁻¹ group had a higher RR interval than the others (Fig. 5 C). The mean PQ interval for the control group was 82.22 ± 3.52 ms, with no significant difference from the vehicle group (p = 0.999). The groups treated with 125 µL.L⁻¹ (94.56 ± 3.84 ms) and 150 µL.L⁻¹ were similar (p = 0.996). The 225 µL.L⁻¹ group (140.0 ± 4.52 ms) showed a greater PQ interval than the other groups (Fig. 5 D). The mean duration of the QRS complex for the control group during induction was 19.33 ± 1.80 ms, similar to the vehicle group and the group treated with 125 µL.L⁻¹ (p = 0.512), but shorter than the other groups. The 125 µL.L⁻¹ group (20.78 ± 1.30 ms) was similar to the 150 µL.L⁻¹ group (p = 0.7813). The group treated with 175 µL.L⁻¹ was similar to the group treated with 200 µL.L⁻¹ (p = 0.2027). The 200 µL.L⁻¹ group (28.44 ± 1.81 ms) was similar to the 225 µL.L⁻¹ group (p = 0.512) (Fig. 5 E). For the control group, the mean QT interval during induction was 211.9 ± 14.94 ms, similar to the vehicle group (p = 0.998). The group treated with 125 µL.L⁻¹ (242.2 ± 10.62 ms) was similar to the 150 µL.L⁻¹ group (p = 0.964). The 225 µL.L⁻¹ group (338.4 ± 10.35 ms) had a longer QT interval compared to the other groups (Fig. 5 F). During the recovery phase following exposure to CLEO at concentrations of 125 µL.L⁻¹, 150 µL.L⁻¹, 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹, reversibility of electrocardiographic alterations was observed (Figs. 6 A, B, C, D, and E). However, this reversibility was slower in groups treated with higher concentrations of CLEO. During recovery, the group treated with 125 µL.L⁻¹ showed results similar to the control group (p = 0.4759) (Fig. 6 A). The fish treated with 150 µL.L⁻¹ exhibited a 90.08% recovery of cardiac function compared to the control group (Fig. 6 B). For the group treated with 175 µL.L⁻¹ of CLEO, the recovery was 86.34% (Fig. 6 C), while the group treated with 200 µL.L⁻¹ showed a recovery of 85.88% (Fig. 6 D). Lastly, the group treated with 225 µL.L⁻¹ also presented an 85.88% recovery (Fig. 6 E). Although the treated fish exhibited a slow reversibility, they maintained a sinus rhythm (Fig. 6 ). During recovery, the control group exhibited a mean heart rate of 95.22 ± 7.77 bpm, similar to the vehicle group and the group treated with 125 µL.L⁻¹ (p = 0.4759). The groups treated with 150 µL.L⁻¹ (85.11 ± 2.26 bpm) were comparable to those treated with 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹ (p = 0.5156) (Fig. 7 A). The amplitude of the QRS complex during recovery for the control group was 1.634 ± 0.238 mV, similar to the other groups (F(6, 56) = 0.5940, p = 0.7338) (Fig. 7 B). The mean RR interval during recovery for the control group was 633.1 ± 47.59 ms, comparable to the vehicle and 125 µL.L⁻¹ groups (p = 0.5850). The groups treated with 150 µL.L⁻¹ (704.9 ± 19.63 ms) were similar to the 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹ groups (p = 0.2349) (Fig. 7 C). The PQ interval during recovery for the control was 82.22 ± 3.528 ms, similar to the vehicle and treated groups with 125 µL.L⁻¹, 150 µL.L⁻¹, and 175 µL.L⁻¹ (p = 0.0914). The groups treated with 200 µL.L⁻¹ and 225 µL.L⁻¹ showed higher means than the control and vehicle groups yet were similar to the other treated groups (p = 0.091) (Fig. 7 D). The duration of the QRS complex during recovery for the control group was 19.33 ± 1.80 ms, similar to the vehicle and groups treated with 125 µL.L⁻¹, 150 µL.L⁻¹, 175 µL.L⁻¹, and 200 µL.L⁻¹ (p = 0.3891). The groups treated with 225 µL.L⁻¹ were similar to the groups treated with 175 µL.L⁻¹ and 200 µL.L⁻¹ (p = 0.3891) (Fig. 7 E). During recovery, the QT interval for the control was 211.9 ± 14.94 ms, similar to the vehicle, 125 µL.L⁻¹, and 150 µL.L⁻¹ groups (p = 0.9957). The groups treated with 175 µL.L⁻¹ (228.7 ± 4.92 ms) were similar to the 200 µL.L⁻¹ and 225 µL.L⁻¹ groups (p = 0.7295) but were higher than the other groups (Fig. 7 After the administration of flumazenil (1 mg/kg, i.p.), there was an increase in latency for the loss of postural reflex behavior across all tested concentrations of CLEO. The group treated with flumazenil followed by immersion in 125 µL.L⁻¹ showed an increase of 263.13% in latency. Similar increases were observed in the groups treated with 150 µL.L⁻¹ (208.02%), 175 µL.L⁻¹ (181.4%), 200 µL.L⁻¹ (223.22%), and 225 µL.L⁻¹ (215.77%), indicating that after flumazenil administration, latencies were significantly prolonged (Fig. 8 A). During the recovery period, the group that received flumazenil exhibited shorter latencies for recovering the postural reflex after treatment with different concentrations of CLEO. The group treated with 125 µL.L⁻¹ showed a decrease in latency for recovery of the postural reflex by 53.99%. Similar reductions were noted in the groups treated with 150 µL.L⁻¹ (48.18%), 175 µL.L⁻¹ (52.10%), 200 µL.L⁻¹ (43.26%), and 225 µL.L⁻¹ (38.99%) (Fig. 8 B). 4 Discussion The results from the chromatogram in Fig. 1 , obtained via gas chromatography, revealed the predominance of three terpenes in the essential oil, with Ar-turmerone as the major component (32.5%), followed by Curcumene (13%) and Farnesene (7.2%). The presence of Ar-turmerone as the primary component has been confirmed by other studies (Zhang et al., 2017 ; Guimarães et al., 2020 ; Zheng et al., 2020 ), and its therapeutic potential is widely recognized (Avanço et al., 2017 ; Kheira et al., 2020). The anesthetic properties of Ar-turmerone are currently being investigated, given its chemical similarity to synthetic anesthetics, such as propofol, commonly used in fish anesthesia. Behavioral parameters observed during both induction and recovery from anesthesia showed a dose-dependent effect, with groups receiving higher doses exhibiting a more rapid loss of postural reflex and a slower reversibility (Dos Santos et al., 2022; Vieira et al., 2023 ; Dos Santos et al., 2024 ). The anesthetic induction induced by turmeric oil can be explained by its terpene-rich composition, particularly the Ar-turmerone, which, due to its structure, can modulate GABA A receptors (Bianchini et al., 2017 ; Kasai et al., 2014 ; Khumpirapang et al., 2018 ). This mechanism is similar to that of anesthetics used in humans (Zhou et al., 2012 ). Our findings suggest that lower doses of the oil are safer for use, as evidenced by the group exposed to the dose of 125 µL.L⁻¹, which showed a recovery of 90.08% in cardiac function, similar to the control group. This contrasts with the study by Saccol et al. ( 2017 ), which used significantly higher doses (200 µL.L⁻¹, 300 µL.L⁻¹, and 500 µL.L⁻¹) and observed anesthesia only in groups exposed to 200 µL.L⁻¹. A reduction in heart rate (HR) was observed in fish exposed to turmeric oil compared to the control group, with a direct relationship between concentration and cardiac depression. These results are consistent with studies showing that menthol administration also reduced cardiac activity in Colossoma macropomum in a dose-dependent manner, with reversibility to baseline levels (Cantanhêde et al., 2021 ). Moreover, several studies have reported a reduction in the cardiac function of Tambaqui when exposed to different anesthetic concentrations derived from essential oils, with complete recovery post-anesthesia (De Souza et al., 2019 ; Barbas et al., 2017 ). The prolongation of QT and RR intervals observed in Colossoma macropomum exposed to turmeric oil, compared to the control group, indicates cardiac alterations related to increased oil concentration. However, these effects did not result in impaired cardiac function, aligning with findings from Cantanhêde et al. ( 2021 ) and Da Costa et al. ( 2022 ). The absence of atrioventricular block (AVB) suggests cardiovascular safety, as noted by Zena et al. ( 2021 ). The analysis of the QRS interval showed an increase in its duration with the increasing dose of turmeric oil, but the anesthetic induction with the minimum dose was similar to the control group, indicating that cardiac contraction was maintained. These findings are corroborated by studies by De Sousa et al. (2019) and Vilhena et al. ( 2022 ). Regarding the evaluation of CLEO in relation to the GABA A-benzodiazepine receptor, it appears that the components of the oil act synergistically with the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). The pre-treatment with flumazenil, a benzodiazepine receptor antagonist, increased the latency for the loss of postural reflex and accelerated recovery, confirming the involvement of GABA A receptors in the anesthesia induced by CLEO. This study demonstrated the anesthetic activity of CLEO at various treatment concentrations, evidencing the reversibility of behavioral and cardiac effects. According to our results, the safe range for short-duration anesthesia appears to be between 125 and 225 µL.L⁻¹, while for deeper anesthesia, we recommend concentrations of 200 to 225 µL.L⁻¹, without significant impairments in cardiac activity. The mechanism of action of CLEO components involves the potentiation of the GABAergic system, as indicated by the action of flumazenil, suggesting that the anesthesia induced by CLEO is mediated by allosteric mechanisms at the GABA A receptors. Declarations Conflict of interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Publisher’s note All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher. Funding The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by Fundação Amazônia de Amparo a Estudos e Pesquisas do Estado do Pará (FAPESPA) and The APC was funded by Pró-Reitoria de Pesquisa e Pós Graduação—PROPESP/UFPA. Author Contribution Conceived and designed the experiments: L.E.C and M.H. Performed the experiments: L.E.C., L.V.d.S., L.L.d.R., G.B.B and M.H. Writing-original draft and editing: all authors. Financial support and administrative support: M.H. All authors have read and agreed to the published version of the manuscript. Acknowledgments The authors would like to acknowledge Fundação Amazônia de Amparo a Estudos e Pesquisas do Estado do Pará (FAPESPA) for their financial support provided of this research, Programa de Pós graduação em Ciências Farmacêuticas (PPGCF) and Programa de Pós graduação em Farmacologia e Bioquímica (PPGFARMABIO). Data availability statement The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding authors. References Abdel-Aziz MFA (2013) Effect of some environmental conditions and protein quality on reproductive and productive performance of red hybrid tilapia (Oreochromis sp.). MSc thesis, Animal Dep., Fac. Agri., Fayoum university, Egypt. Aggarwal B, Harikumar K (2009) Potential therapeutic effects of curcumin, the antiinflammatory agent, against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplastic diseases. Int. J. Biochem. Cell Biol, 41(1), 40–59. https://doi.org/10.1016/j.biocel.2008.06.010 . Arnaout R, Ferrer T, Huisken J, Spitzer K, Stainier DYR, Tristani-Firouzi M, et al (2007) “Zebrafish model for human long QT syndrome,” Proceedings of the National Academy of Sciences, vol. 104, p. 11316. doi: 10.1073/pnas.0702724104 . Avanço GB, Ferreira FD, Bomfim NS, de Souza Rodrigues dos Santos PA, Peralta RM, Brugnari T, Mallmann CA, de Abreu Filho BA, Mikcha JMG, Machinski M (2017) Curcuma longa L. essential oil composition, antioxidant effect, and effect on Fusarium verticillioides and fumonisin production. Food Control 73:806–813. doi: 10.1016/j.foodcont.2016.09.032 . Aydın I, Akbulut B, Kucuk, E, Kumlu M (2015) Effects of temperature, fish size and dosage of clove oil on anaesthesia in turbot (Psetta maxima Linnaeus). Journal of Fisheries and Aquatic Science. Sci, 15, 899–904. doi: 10.4194/1303-2712-v15_4_13 . Barbas LAL, Hamoy M, de Mello VJ, Barbosa RPM, de Lima HST, Torres MF, Gomes, MRF (2017) Essential oil of citronella modulates electrophysiological responses in tambaqui Colossoma macropomum: a new anaesthetic for use in fish. Aquaculture, 479, 60–68. https://doi.org/10.1016/j.aquaculture.2017.05.027 . Barbas LAL, Maltez LC, Stringhetta GR, de Oliveira GL, Monserrat JM, da Silva DT, Sampaio LA (2017) Properties of two plant extractives as anaesthetics and antioxidants for juvenile tambaqui Colossoma macropomum. Aquaculture, 469, 79–87. https://doi.org/10.1016/j.aquaculture.2016.12.012 . Barbas LAL, Stringhetta GR, de Oliveira GL, Figueiredo MRC, Sampaio LA (2016) Jambu, Spilanthes acmella as a novel anaesthetic for juvenile tambaqui, Colossoma macropomum: secondary stress responses during recovery. Aquaculture, 456, 70–75. https://doi.org/10.1016/j.aquaculture.2016.01.026 . Becker AG, Parodi TV, Heldwein CG, Zeppenfeld CC, Heinzmann BM, Baldisserotto B (2012) Transportation of silver catfish, Rhamdia quelen, in water with eugenol and the essential oil of Lippia alba. Fish Physiol. Biochem. 38, 789–796. doi: 10.1007/s10695-011-9562-4 . Bianchini AE, Garlet QI, Da Cunha JA, Bandeira Junior G, Brusque ICM, Salbego J, Heinzmann BM, Baldisserotto B (2017) Monoterpenoids (Thymol, carvacrol and S-(+)- linalool) with anesthetic activity in silver catfish (Rhamdia quelen): Evaluation of acetylcholinesterase and GABaergic activity. Brazilian J. Med. Biol. Res. 50, 1–8. https://doi.org/10.1590/1414-431x20176346 . Cantanhêde SM, Amado LL, da Costa BMPA, Barbas LAL, Torres MF, Hamoy AO, Hamoy M (2021) Menthol exposure induces reversible cardiac depression and reduces lipid peroxidation in the heart tissue of tambaqui Colossoma macropomum. Aquaculture, 541, 736847. https://doi.org/10.1016/j.aquaculture.2021.736847 Cao Y, Yan H, Yu G, Su R (2019) Flumazenil-insensitive benzodiazepine binding sites in GABA A receptors contribute to benzodiazepine-induced immobility in zebrafish larvae. Life Sci. 15;239:117033. doi: 10.1016/j.lfs.2019.117033 da Costa, BMA, Torres, MF, da Silva RA, Aydın B, Amado LL, Hamoy M, Barbas LAL (2022) Integrated behavioural, neurological, muscular and cardiorespiratory response in tambaqui, Colossoma macropomum anaesthetized with menthol. Aquaculture, 560, 738553. https://doi.org/10.1016/j.aquaculture.2022.738553 Davis MW (2010) Fish stress and mortality can be predicted using reflex impairment. Fish and Fisheries, 11(1), 1–11. https://doi.org/10.1111/j.1467-2979.2009.00331.x de Souza, ADSL, Peret AC, Hamoy M, de Souza RAL, Torres MF, Barbas LAL (2019) Propofol and essential oil of Nepeta cataria induce anaesthesia and marked myorelaxation in tambaqui Colossoma macropomum: implications on cardiorespiratory responses. Aquaculture, 500,160–169. https://doi.org/10.1016/j.aquaculture.2018.10.017 Delbon MC, Paiva MR (2012) Eugenol in tilapia juvenile: concentrations and successive administrations. Boletim do Instituto de Pesca, 38(1): 43–52. Dos Santos MF, do Nascimento LM, da Paz CA, Câmara TM, Motomya YKM, da Cunha Ferreira R, da Silva Deiga Y, Monteiro E, Cantanhêde SM, Amado LL, Hamoy M (2024) Behavioral and electrophysiological study in Colossoma macropomum treated with different concentrations of Nepeta cataria oil in an immersion bath revealed a therapeutic window for anesthesia. Fish Physiol Biochem. Aug;50(4):1651–1665. doi: 10.1007/s10695-024-01361-2 Faleschini T, Syafni N, Schulte HL, Garifulina A, Hering S, Espindola LS, Hamburger M (2023) A neolignan from Connarus tuberosus as an allosteric GABA A receptor modulator at the neurosteroid binding site. Biomed Pharmacother. 2023;161:114498. doi: 10.1016/j.biopha.2023.114498 FAO (2014) Food and Agriculture Organization of the United Nations.El estado mundial de la pesca y la acuicultura. Oportunidades y desafíos. 274p. Felix LM, Luzio A, Antunes L, Coimbra AM, Valentim AM (2021) Malformations and mortality in zebrafish early stages associated with elevated caspase activity after 24 h exposure to MS-222. Toxicology and Applied Pharmacology, 412, Article 115385. https://doi.org/10.1016/j.taap.2020.115385 Genge CE, Lin E, Lee L, Sheng X, Rayani K, Gunawan M, Stevens CM, Li AY, Talab SS, Claydon TW, Hove-Madsen L, Tibbits GF (2016) Reviews of Physiology, Biochemistry and Pharmacology, vol. 171. Springer International Publishing, Cham, pp. 99–136. Goulding M, Carvalho M (1982) Life history and management of the tambaqui (Colossoma macropomum, Characidae): an important Amazonian food fish. Revista Brasileira de Zoologia, 1, 107–133. doi: 10.1590/S0101-81751982000200001 Guilherme HO, Santos FAC, Costa LS, Prado VGL, Palheta GDA, de Melo NFAC, Luz RK, Ribeiro PAP (2022) Feeding, growth, and blood chemistry of the tambaqui (Colossoma macropomum) held under self-feeding and time-restricted automatic feeding conditions. Aquac. Int. 30, 323–339. https://doi.org/10.1007/s10499-021-00801-7 Guimarães IG, Miranda EC, Araújo JG (2014) Coefficients of total tract apparent digestibility of some feedstuffs for Tambaqui (Colossoma macropomum). Animal Feed Science and Technology, 188, 150–155. https://doi.org/10.1016/j.anifeedsci.2013.11.007 Guimarães AFAC, Vinhas AF, Gomes LH, Souza PBK (2020) Essential oil of Curcuma longa Rhizomes chemical composition, yield variation stability. Quim. Nova, Vol. 43, No. 7, 909–913. doi: 10.21577/0100-4042.20170547 Hamoy AO, Fonseca SMD, Cei GL, Júnior FLDA, Hamoy MKO, Ribeiro RM, Barbas LAL, Muto N, Hamoy M (2023) Behavioral, electrocorticographic and electrocardiologic changes in Colossoma macropomum (Tambaqui) in the effect of cunaniol. PLoS One. Jun 30;18(6):e0287681. doi: 10.1371/journal.pone.0287681 Harmon TS, (2009) Methods for reducing stressors and maintaining water quality associated with live fish transport in tanks: a review of the basics. Reviews in Aquaculture, 1, 58–66. https://doi.org/10.1111/j.1753-5131.2008.01003.x Hasan H, Farida EG (2016) Different concentration of rubber seed extract (Hevea brasiliensi) for anesthetic to the survival of prospective broodstock milkfish by using closed transportation. Jurnal Ruaya, 4(2),55–62. https://doi.org/10.1016/j.aaf.201903002 Hohlenwerger JC, Baldisserotto B, Couto RD, Heinzmann BM, da Silva DT, Caron BO, Schmidt D, Copatti CE (2017) Essential oil of Lippia alba in the transport of Nile tilapia. Ciência Rural, 47(3). https://doi.org/10.1590/0103-8478cr20160040 Hoseini SM, Mirghaed AT, Yousefi M (2019) Application of herbal anaesthetics in aquaculture. Reviews in Aquaculture, 11, 550–564. https://doi.org/10.1111/raq.12245 IBGE (2020) Instituto Brasileiro de Geografia e Estatística – Produção de aquicultura. http://www.sidra.ibge.gov.br/bda/tabela/listabl.asp?c=3940&z=t&o=21 . (Accessed 19.09.2024). Kasai M, Hososhima S, Yun- Fei L (2014) Menthol induces surgical anesthesia and rapid movement in fishes. The Open Neuroscience Journal, 8, 1–8. https://doi.org/10.2174/1874082001408010001 Kheira M, Boumediene M, Aicha TT, Pascal S (2023) Chemical Composition and Antioxidant and Antimicrobial Activities of Curcuma longa L. Essential Oils. Egyptian Journal of Chemistry 66(7): 283–295. doi: 10.21608/ejchem.2022.121586.5555 Khumpirapang N, Chaichit S, Jiranusornkul S, Pikulkaew S, Mullertz A, Okonogi S (2018) In vivo anesthetic effect and mecha nism of action of active compounds from Alpinia galanga oil on Cyprinus carpio (koi carp). Aquaculture, 496, 176–184. https://doi.org/10.1016/j.aquaculture.2018.07.017 Kim HT, Lee JH, Park IB, Heo HE, Kim TY, Lee MJ (2010) Long QT syndrome provoked by induction of general anesthesia-A case report. Korean Journal of Anesthesiology, 59(Suppl), S114-S118. https://doi.org/10.4097/kjae.2010.59.S.S114 Koopman CD, De Angelis J, Iyer SP, Verkerk AO, Da Silva J, Berecki G, Jeanes A, Baillie GJ, Paterson S, Uribe V, Ehrlich OV, Robinson SD, Garric L, Petrou S, Simons C, Vetter I, Hogan BM, de Boer TP, Bakkers J, Smith KA (2021) The zebrafish grime mutant uncovers an evolutionarily conserved role for Tmem161b in the control of cardiac rhythm. Proc Natl Acad Sci USA.;11:8. doi: 10.1073/pnas.2018220118 Le T, Zhang J, Nguyen AH, Trigo Torres RS, Vo K, Dutt N, Lee J, Ding Y, Xu X, Lau MPH, Cao H (2022) A novel wireless ECG system for prolonged monitoring of multiple zebrafish for heart disease and drug screening studies. Biosens Bioelectron. Feb 1;197:113808. doi: 10.1016/j.bios.2021.113808 Luz JGR, Nogueira JN, Alves CMG, Videira MN, Canuto KM, Castro KNC, Tavares-Dias M (2021) Essential oil of Alpinia zerumbet (Zingiberaceae) has anthelmintic efficacy against monogenean of Colossoma macropomum (Characiformes: Serrasalmidae). Aquac. Res. 52, 5340–5349. https://doi.org/10.1111/are.15404 McGrath M, Hoyt H, Pence A, Forman SA, Raines DE (2021) Selective actions of benzodiazepines at the transmembrane anaesthetic binding sites of the GABA A receptor: In vitro and in vivo studies. Br J Pharmacol. Dec;178(24):4842–4858. doi: 10.1111/bph.15662 Merola N, Cantelmo AO (1987) Growth, feed conversion and mortality of cagereared tambaqui, Colossoma macropomum, fed various dietary feeding regimes and protein levels. Aquaculture, 66(3–4), 223–233. https://doi.org/10.1016/0044-8486(87)90108-6 ; Nakayama CL, Silva LFS, dos Santos FAC, Boaventura TP, Favero GC, Palheta GDA, Melo NFAC, Romano LA, Luz RK (2022) Zootechnical performance and some physiological indices of tambaqui, Colossoma macropomum juveniles during biofloc maturation and in different feed regimes. Agriculture 12, 1025. https://doi.org/10.3390/agriculture12071025 Parveen Z, Nawaz S, Siddique S, Shahzad K (2013) Composition and antimicrobial activity of the essential oil from leaves of Curcuma longa L. Kasur Variety. Indian Journal of Pharmaceutical Sciences, 75(1), 117–122. https://dx.doi.org/10.4103%2F0250-474X.113544 PEIXE BR (2024). Anuário Peixe-BR da piscicultura 2024. São Paulo. Priborsky J, Velisek J (2018) A review of three commonly used fish anesthetics. Rev. Fish. Sci. Aquac. 26, 417–442. doi: 10.1080/23308249.2018 . 1442812 Primadona R, Lestari S, Baehaki A (2017) The effect of electric current on survival rate of fry finger (Clarias sp.) in dry transportation. Fistech-Jurnal Teknologi Hasil Perikanan, 6(2): 145–152. https://doi.org/10.36706/fishtech.v6i2.5846 Priya R, Prathapan A, Raghu KG, NirmalaMenon A (2012) Chemical composition and in vitro antioxidative potential of essential oil isolated from Curcuma longa L. leaves. Asian Pac J Trop Biomed. S695–9. Ramanayaka JC, Atapattu NSBM (2006) Fish Anaesthetic properties of some local plant material. Tropical Agricultural Research and Extension, 9,1–6. doi: 10.4038/tare.v9i0.5413 Rezende FP, Pascoal LM, Vianna RA, Lanna EAT (2017) Sedation of nile tilapia with essential oils: Tea tree, clove, Eucalyptus, and mint oils. Rev. Caatinga, Mossoró. 30(2), 479–486. https://doi.org/10.1590/1983-21252017v30n223rc Ribeiro AS, Batista ES, Dairiki JK, Chaves FCM, Inoue LA (2016) Anesthetic properties of Ocimum gratissimum essential oil for juvenile matrinxã. Acta Sci. Anim. Sci. 38, 1–7. https://doi.org/10.4025/actascianimsci.v38i1.28787 Saccol EMH, Toni C, Pess TS, Ourique GM, Gressler LT, Silva LVF, Mourão RHV, Oliveira RB, Baldisserotto B, Pavanato MA (2017) Anesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils oils on tambaqui (Colossoma macropomum). Aquac. Res. 48, 2012–2031. doi: 10.1111/are.13034 Schramm A, Ebrahimi SN, Raith M, Zaugg J, Rueda DC, Hering S, Hamburger M (2013) Phytochemi- cal profiling of Curcuma kwangsiensis rhizome extract, and identification of labdane diterpenoids as positive GABAA receptor modulators. Phytochemistry 96, 318–329. doi: 10.1016/j.phytochem.2013.08.004 Suwandi R, Nugraha R, Zulfamy KE (2013) Aplikasi ekstrak daun jambu Psidium guajava var.pomifera pada proses transportasi ikan nila. Oreochromis niloticus) JPHPI,16(1), 69–78. https://doi.org/10.17844/jphpi.v16i1.8107 Teixeira RR, de Souza RC, Sena AC, Baldisserotto B, Heinzmann BM, Copatti CE (2018) Essential oil of Aloysia Triphylla is effective in Nile Tilapia transport. Bol. Inst. Pesca 44, 17–24. https://doi.org/10.20950/1678-2305.2018.263 Vieira LR, Pereira YLG, Diniz LA, Nascimento CP, Da Silva ALM, Azevedo JEC, De Mello VJ, Muto NA, Barbas LAL, Hamoy M (2023) Graded concentrations of lidocaine hydrochloride in the modulation of behavioral, cardiac, and muscular responses of the Amazon freshwater fsh tambaqui (Colossoma macropomum). Aquaculture. 563:738985. https://doi.org/10.1016/j.aquaculture.2022.738985 Vieira ELP, Egger RC, Lima TC, Paiva MB, Gonçalves LU, Honczaryk A, Godoy L, Tavares GC, Kuradomi RY (2022) Short-term preservation of Colossoma macropomum oocytes by different methods and chilling temperatures. Theriogenology 191, 22–34. https://doi.org/10.1016/j Vijayasteltar BL, Kottarapat J, Ramadasan K (2011) An evaluation of antioxidant, anti-inflammatory, and antinociceptive activities of essential oil from Curcuma longa L. Indian Journal of pharmaceutical Sciences, 43(5), 526–531. https://dx.doi.org/10.4103%2F0253-7613.84961 Vilhena CS, da Silva RA, da Costa BMPA, Torres MF, de Mello VJ, Noronha RCR, do Rosário da Silva JK, Hamoy M, Barbas LAL, do Nascimento LAS (2022) Cardiac response in tambaqui Colossoma macropomum anaesthetised with Piper divaricatum essential oil. Fish Physiol Biochem. 48(5):1413–1425. doi: 10.1007/s10695-022-01132-x Wolkers CP, Barbosa Junior A, Menescal-de-Oliveira L, Hoffmann A (2015) GABAA-benzodiazepine receptors in the dorsomedial (Dm) telencephalon modulate restraint-induced antinociception in the fish Leporinus macrocephalus. Physiol Behav.Aug 1;147:175 – 82. doi: 10.1016/j.physbeh.2015.04.037 Yu ZF, Kong LD, Chen Y (2002) Antidepressant activity of aqueous extracts of Curcuma longa in mice. Journal of Ethnopharmacology 83, 161–165. doi: 10.1016/s0378-8741(02)00211-8 Zahl IH, Samuelsen O, Kiessling A (2012) Anaesthesia of farmed fish: implications for welfare. Fish Phys- iol Biochem 38:201–218. https://doi.org/10.1007/s10695-011-9565-1 Zena LA, Ekström A, Gräns A, Olsson C, Axelsson M, Sundh H, Sandblom E (2021) It takes time to heal a broken heart: ventricular plasticity improves heart performance after myocardial infarction in rainbow trout, Oncorhynchus mykiss. J Exp Biol. Dec 1;224(23):jeb243578. doi: 10.1242/jeb.243578 Zhang L, Yang Z, Chen F, Su P, Chen D, Pan W (2017) Composition and bioactivity assessment of essential oils of Curcuma longa L. collected in China. Ind Crops Prod.;109(15):60–73. Zheng Y, Pan C, Zhang Z, Luo W, Liang X, Shi Y, Liang L, Zheng X, Zhang L, Du Z (2020) Antiaging effect of Curcuma longa L. essential oil on ultravioletirradiated skin. Microchemical Journal 154.104608. https://doi.org/10.1016/j.microc.2020.104608 Zhou C, Liu J, Chen XD (2012) General anesthesia mediated by effects on ion channels. World Journal of Critical Care Medicine, 1, 80–93. https://doi.org/10.5492/wjccm.v1.i3.80 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5154092","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":370928660,"identity":"0cfb6cfd-169f-4dbe-8e1e-1a81dc86e63e","order_by":0,"name":"Luciana Esquerdo Cerqueira","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Luciana","middleName":"Esquerdo","lastName":"Cerqueira","suffix":""},{"id":370928661,"identity":"50ef876c-ef72-450b-8a58-b335568f0d7f","order_by":1,"name":"Daniella Bastos de Araújo","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1klEQVRIiWNgGAWjYDCCAyBkwMDA3t4A5BlYkKCF58wBkBYJ4rSAAc+NBBBFhBa+42cPHuYpOCzPI/n86oYfBRIM/O3dCXi1SJ7JSzjMY3DYsEc6p+xmD9BhEmfObsCrxeBAjsHBGQZpjPulc9Ju8AC1GEjkEtBy/g1Yi32P5Jm0m3+I0nIjx+DABwObxB4J9mO3ibJF8sYbsJbkHp4cttsyBhI8BP3Cdz7H+EPCHwnbHvbjz26++WMjx9/ei18LEuAxAJPEKgcB9gekqB4Fo2AUjIIRBABOKEtjW2VFkQAAAABJRU5ErkJggg==","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":true,"prefix":"","firstName":"Daniella","middleName":"Bastos","lastName":"de Araújo","suffix":""},{"id":370928663,"identity":"a7628d9a-bdaf-49d4-9033-0427d65a4b97","order_by":2,"name":"Luciana Eiró-Quirino","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Luciana","middleName":"","lastName":"Eiró-Quirino","suffix":""},{"id":370928664,"identity":"089c140c-be3a-45ef-a1f4-043366b62c21","order_by":3,"name":"Priscille Fidelis Pacheco Hartcopff","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Priscille","middleName":"Fidelis Pacheco","lastName":"Hartcopff","suffix":""},{"id":370928665,"identity":"35097ed5-4bdb-4af0-aa7b-dd7781bac88b","order_by":4,"name":"Clarissa Araújo da Paz","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Clarissa","middleName":"Araújo da","lastName":"Paz","suffix":""},{"id":370928666,"identity":"1b5e3f67-6fae-44c6-a506-28b744e651ad","order_by":5,"name":"Thaysa Sousa Reis","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Thaysa","middleName":"Sousa","lastName":"Reis","suffix":""},{"id":370928667,"identity":"78065c62-b1b2-4847-b218-c4f0550e8bb4","order_by":6,"name":"Sarah Farias Camara","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Sarah","middleName":"Farias","lastName":"Camara","suffix":""},{"id":370928668,"identity":"ea06e30d-ff15-4712-8ca1-0e5615a77d19","order_by":7,"name":"Luana Vasconcelos de Souza","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Luana","middleName":"Vasconcelos","lastName":"de Souza","suffix":""},{"id":370928669,"identity":"b1093fe1-a40f-4601-94f2-731b225a9bc1","order_by":8,"name":"Lucas Lima Rocha","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Lucas","middleName":"Lima","lastName":"Rocha","suffix":""},{"id":370928670,"identity":"6b110d77-4210-4e62-b856-49c23c3f1dd3","order_by":9,"name":"Gabriela Brito Barbosa","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Gabriela","middleName":"Brito","lastName":"Barbosa","suffix":""},{"id":370928671,"identity":"40e3b878-0ff5-469b-ba8f-4d0dc61d51cd","order_by":10,"name":"Rômulo Augusto Feio Farias","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Rômulo","middleName":"Augusto Feio","lastName":"Farias","suffix":""},{"id":370928672,"identity":"361c5e60-e4f1-48c3-b035-7cacffdf2d22","order_by":11,"name":"Nilton Akio Muto","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Nilton","middleName":"Akio","lastName":"Muto","suffix":""},{"id":370928673,"identity":"29c72f48-6ca3-4057-b085-d20cb32afcaa","order_by":12,"name":"Moisés Hamoy","email":"","orcid":"","institution":"Federal University of Pará (UFPA)","correspondingAuthor":false,"prefix":"","firstName":"Moisés","middleName":"","lastName":"Hamoy","suffix":""}],"badges":[],"createdAt":"2024-09-25 19:08:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5154092/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5154092/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":69012116,"identity":"0c77f860-4b81-493f-bb07-15c1380535ca","added_by":"auto","created_at":"2024-11-14 14:02:17","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":18981,"visible":true,"origin":"","legend":"\u003cp\u003eChromatogram of the \u003cem\u003eCurcuma longa\u003c/em\u003e essential oil sample. Performed at the Chromatography Laboratory, Department of Chemistry, Central Analysis Facility – Federal University of Minas Gerais, Avenida Antônio Carlos, 6627 - Campus Pampulha - Belo Horizonte, MG, Brazil\u003c/p\u003e","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-5154092/v1/dcc3bcf0851e6683088a033f.png"},{"id":69012117,"identity":"474020f7-cf47-47a1-9d6d-8de6fdbc1989","added_by":"auto","created_at":"2024-11-14 14:02:17","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":45535,"visible":true,"origin":"","legend":"\u003cp\u003eMean latencies (in seconds) for the loss of posture reflex during immersion baths with different CLEO treatments (A). Recovery of the posture reflex after exposure to various CLEO concentrations (B). (ANOVA followed by Tukey's test; *p\u0026lt;0.05, **p\u0026lt;0.01, and ***p\u0026lt;0.001)\u003c/p\u003e","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-5154092/v1/b29a599a8999320550e96134.png"},{"id":69012120,"identity":"4376343d-ac39-4c89-a556-01f0704e589d","added_by":"auto","created_at":"2024-11-14 14:02:17","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":53145,"visible":true,"origin":"","legend":"\u003cp\u003eNormal electrocardiographic (ECG) recording of tambaqui, \u003cem\u003eColossoma macropomum\u003c/em\u003e, lasting 5 minutes (A). Expanded electrocardiogram during the final 30 seconds of the recording (270-300 s) (B). Amplification of the last 5 seconds of the recording, highlighting morphological elements such as the P wave, QRS complex, and T wave, along with analyzed parameters: heart rate (bpm), amplitude of the QRS complex (mV), P-Q interval (ms), R-R interval (ms), QRS duration (ms), and Q-T interval (ms) (C)\u003c/p\u003e","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-5154092/v1/776da3bc0435887d67057a97.png"},{"id":69013162,"identity":"3bca96bb-5486-4783-8ee3-33da9f65619c","added_by":"auto","created_at":"2024-11-14 14:18:17","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":174591,"visible":true,"origin":"","legend":"\u003cp\u003eElectrocardiographic (ECG) tracings demonstrating cardiac activity in juvenile \u003cem\u003eColossoma macropomum\u003c/em\u003e (left), amplification of the last 30 seconds of the 5-minute recording (270-300 s) (center), illustrating the graph elements evaluated in the study, including the P wave, QRS complex, and T wave (5-second amplification) (right). Cardiac activity in juveniles of \u003cem\u003eColossoma macropomum\u003c/em\u003e during immersion treatment with the vehicle, detailing the cardiac graph elements (A). Groups treated with immersion in different concentrations of CLEO, with amplification and identification of cardiac deflections: 125 µL.L⁻¹ (B), 150 µL.L⁻¹ (C), 175 µL.L⁻¹ (D), 200 µL.L⁻¹ (E), and 225 µL.L⁻¹ (F)\u003c/p\u003e","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-5154092/v1/9178357ec9abf83e2c68cc56.png"},{"id":69012953,"identity":"9427c18e-f2fe-4d83-b519-c19ce66f95cc","added_by":"auto","created_at":"2024-11-14 14:10:17","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":86391,"visible":true,"origin":"","legend":"\u003cp\u003eMean values of heart rate in beats per minute (bpm) (A), mean amplitude of the QRS complex (mV) (B), mean RR intervals (ms) (C), mean PQ interval (ms) (D), duration of the QRS complex (ms) (E), and QT interval (ms) (F) during exposure to CLEO at concentrations of 125 µL.L⁻¹, 150 µL.L⁻¹, 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹ (ANOVA followed by Tukey's test; *p \u0026lt; 0.05, **p \u0026lt; 0.01, ***p \u0026lt; 0.001; n = 9)\u003c/p\u003e","description":"","filename":"Onlinefloatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-5154092/v1/0261db27e55dfe5a0d37a2e5.png"},{"id":69013163,"identity":"31cf9371-20fd-42ff-b421-dba08e749937","added_by":"auto","created_at":"2024-11-14 14:18:17","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":161390,"visible":true,"origin":"","legend":"\u003cp\u003eCardiac activity in juveniles of \u003cem\u003eColossoma macropomum\u003c/em\u003e during recovery following immersion baths with different concentrations of CLEO (left). Amplification of the recording over the last 30 seconds (270-300 s) for the identification of cardiac deflections (center). Recovery period after immersion baths with the following concentrations of CLEO (amplification of 5 s) (right): 125 µL.L⁻¹ (A), 150 µL.L⁻¹ (B), 175 µL.L⁻¹ (C), 200 µL.L⁻¹ (D), and 225 µL.L⁻¹ (E)\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-5154092/v1/052989e1a79535ef97a787c7.png"},{"id":69012955,"identity":"93a72ec0-9661-4e4d-a935-14bcb890b343","added_by":"auto","created_at":"2024-11-14 14:10:17","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":84176,"visible":true,"origin":"","legend":"\u003cp\u003eAverage values of cardiac parameters during recovery from treatment with different concentrations of CLEO in immersion baths at 125 µL.L⁻¹, 150 µL.L⁻¹, 175 µL.L⁻¹, 200 µL.L⁻¹, and 225 µL.L⁻¹. The figure includes the average heart rate (bpm) (A), QRS amplitude (mV) (B), R-R intervals (ms) (C), P-Q intervals (ms) (D), duration of the QRS complex (ms) (E), and QT interval (ms) (F). Data were analyzed using ANOVA followed by Tukey's test; *P\u0026lt;0.05, **P\u0026lt;0.01, ***P\u0026lt;0.001; n=9\u003c/p\u003e","description":"","filename":"Onlinefloatimage8.png","url":"https://assets-eu.researchsquare.com/files/rs-5154092/v1/abbb574d956cd73ca0e4946d.png"},{"id":69012113,"identity":"06d1cc8b-f0f6-4a0a-abe7-e9a9ed5cb5ce","added_by":"auto","created_at":"2024-11-14 14:02:17","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":20412,"visible":true,"origin":"","legend":"\u003cp\u003eCardiac activity in juveniles of \u003cem\u003eColossoma macropomum\u003c/em\u003e during recovery following immersion baths with different concentrations of CLEO (left). Amplification of the recording over the last 30 seconds (270-300 s) for the identification of cardiac deflections (center). Recovery period after immersion baths with the following concentrations of CLEO (amplification of 5 s) (right): 125 µL.L⁻¹ (A), 150 µL.L⁻¹ (B), 175 µL.L⁻¹ (C), 200 µL.L⁻¹ (D), and 225 µL.L⁻¹ (E)\u003c/p\u003e","description":"","filename":"Onlinefloatimage9.png","url":"https://assets-eu.researchsquare.com/files/rs-5154092/v1/8773378fffe0e0c77ec555e7.png"},{"id":72487334,"identity":"7dda6de8-b7ad-4a84-bf07-ffc58c4b55f2","added_by":"auto","created_at":"2024-12-27 19:31:33","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1514652,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5154092/v1/5d79d174-d20a-4b12-9e4a-682adcc86ffe.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Behavioral, electrocardiographic evaluation, and mechanism of action of Curcuma longa essential oil in juvenile tambaqui Colossoma macropomum (Cuvier, 1818) subjected to immersion baths at different concentrations","fulltext":[{"header":"1 Introduction","content":"\u003cp\u003eAquaculture activities subject fish to various potential stressors. Handling and transport procedures, for instance, can result in injuries that serve as entry points for infections and bacteria (Ribeiro et al., \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Barbas et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Primadona et al., \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Additionally, high stocking densities in aquaculture environments increase stress levels in fish (Zahl et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), which significantly impacts their physiology and survival (Harmon, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Stress contributes to elevated fish mortality rates (Delbon \u0026amp; Paiva, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Abdel-Aziz, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), potentially leading to significant resource and productivity losses in the production chain (Davis, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Becker et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Hohlenwerger et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Teixeira et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). As such, anesthetic strategies have become crucial in ensuring the success of aquaculture by promoting animal welfare during and after handling procedures (Bianchini et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Hoseini et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAmong anesthetics used in fish, those of natural origin have been widely studied due to their efficacy in transport when administered in low doses to induce anesthesia (Suwandi, Nugraha, \u0026amp; Zulfamy, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Aydın et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Additionally, they increase post-administration survival rates (Hasan et al., 2016), exhibit low tissue residue in animals, and reduce the number of pollutants released into the environment (Ramanayaka \u0026amp; Atapattu, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Rezende, Pascoal, Vianna, \u0026amp; Lanna, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eOne such natural product is \u003cem\u003eCurcuma longa\u003c/em\u003e essential oil (CLEO), which has demonstrated potential for use in fish anesthesia. According to Schramm et al. (\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), certain constituents of this species may exert pharmacological effects possibly associated with the GABAergic system. \u003cem\u003eCurcuma longa\u003c/em\u003e is well-known for its diverse pharmacological and therapeutic properties (Aggarwal et al., 2009), including antioxidant, anticancer, antiatherosclerotic, antidiabetic, anti-inflammatory, immunomodulatory, and sedative activities (Yu, Kong \u0026amp; Chen, \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Vijayasteltar et al., \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Priya et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Parveen et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eTambaqui\u003c/em\u003e (\u003cem\u003eColossoma macropomum\u003c/em\u003e) is the most widely farmed native fish species in Brazil and across several nations in South and Central America, according to reports from FAO (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) and IBGE (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Belonging to the family Serrasalmidae, \u003cem\u003etambaqui\u003c/em\u003e is endemic to the Amazon and Orinoco River basins (Merola, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1987\u003c/span\u003e; Goulding, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e1982\u003c/span\u003e). This Amazonian species exhibits high growth rates and feed conversion efficiency, as demonstrated by Merola and Cantelmo (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1987\u003c/span\u003e). The growing interest in \u003cem\u003etambaqui\u003c/em\u003e aquaculture can be attributed to its high meat quality, growth rate, and feed efficiency (Guimar\u0026atilde;es et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Furthermore, studies by Barbas et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2016\u003c/span\u003ea, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2016\u003c/span\u003eb) suggest that \u003cem\u003etambaqui\u003c/em\u003e is a suitable model for anesthetic studies in tropical freshwater species.\u003c/p\u003e \u003cp\u003eAccording to data from the Brazilian Aquaculture Association (2024), Brazil produced approximately 887,029 tons of native fish in 2023, with \u003cem\u003etambaqui\u003c/em\u003e being the second most exported species, demonstrating an 809% increase in production (PEIXE BR, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) compared to the previous year. This growing profitability has spurred interest in developing more efficient farming strategies to enhance productivity, as well as the health and welfare of these fish (Guilherme et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Luz et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Nakayama et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Vieira et al., \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe concern for fish welfare is highly relevant in anesthesia studies. Electrocardiography (ECG) is one of the tools used to ensure that vital functions are not compromised during anesthesia. Studies by Arnaout et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2007\u003c/span\u003e) and Koopman et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) noted the similarity between the ECG waveforms (P waves, QRS complexes, and T waves) of adult zebrafish and humans. Additionally, zebrafish studies (Genge et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Le et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) have been successful in screening drug-induced cardiac damage, including drug-induced QT interval prolongation.\u003c/p\u003e \u003cp\u003eThus, this study evaluates the behavioral and electrocardiographic responses of \u003cem\u003etambaqui\u003c/em\u003e (\u003cem\u003eColossoma macropomum\u003c/em\u003e) subjected to immersion baths with varying concentrations of \u003cem\u003eCurcuma longa\u003c/em\u003e essential oil, aiming to ensure the safe use of this oil in anesthetic induction and recovery.\u003c/p\u003e"},{"header":"2 Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Experimental Animals\u003c/h2\u003e \u003cp\u003eThe experimental subjects used in this study were \u003cem\u003eColossoma macropomum\u003c/em\u003e (tambaqui) (n\u0026thinsp;=\u0026thinsp;108, males and females). The fish were housed in aquaria at the Experimental Facility of the Laboratory of Pharmacology and Toxicology of Natural Products at the Federal University of Par\u0026aacute; (ICB/UFPA) under controlled temperature conditions (25\u0026ndash;27\u0026deg;C) and a 12-hour light/12-hour dark photoperiod. The animals were fed commercial feed (32% protein) twice daily to satiation. Unconsumed feed and feces were removed by siphoning, and approximately 30% of the tank water was renewed daily with filtered water from the same source. During a 10-day acclimatization period, water quality parameters were monitored, including water temperature (26.3\u0026deg;C) and pH (7.6). The research project was submitted to and approved by the UFPA Research Ethics Committee file number 9261280624 [ID 002674].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Organoleptic Properties and Chromatographic Analysis\u003c/h2\u003e \u003cp\u003e \u003cem\u003eCurcuma longa\u003c/em\u003e essential oil (CLEO) was purchased from Laszlo Aromatherapy Laboratory (Brazil, CNPJ: 07.997.093/0001\u0026ndash;10). The oil was extracted by steam distillation, and chromatographic analysis was performed using Gas Chromatography-Mass Spectrometry (GC-MS) on an AGILENT 7820A GC system under the following conditions: HP-5 column (30 m x 0.32 mm x 0.25 \u0026micro;m; AGILENT), column temperature: 70\u0026deg;C (0 min), 3\u0026deg;C/min up to 250\u0026deg;C, injector temperature: 250\u0026deg;C, split ratio: 1:50, detector temperature: FID 260\u0026deg;C, injection volume: 1 \u0026micro;L (1% in chloroform). The phytoconstituents of the oil are listed in Table\u0026nbsp;1, with the major component being ar-turmerone (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eTable I\u003c/b\u003e Composition of \u003cem\u003eCurcuma longa\u003c/em\u003e essential oil (CLEO)\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Taba\" border=\"1\"\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRetention time order\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIdentification\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCareno\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1,8-Cineol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFarneseno\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e7.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eα-curcumeno\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eβ-curcumeno\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eα-bergamoteno\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eβ-bisaboleno\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSesquifelandreno\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCurlona\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ear-turmerona\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e32.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTurmerol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAtlantona\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Experimental Design\u003c/h2\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e2.3.1 Experiment with Curcuma longa Essential Oil (CLEO)\u003c/h2\u003e \u003cp\u003eJuvenile tambaqui (\u003cem\u003eColossoma macropomum\u003c/em\u003e) weighing 18.13\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1 g were randomly distributed into the following treatments: a) Control; b) Vehicle group (fish subjected to an immersion bath in 3 mL of 70% alcohol diluted in 1 liter of aquarium water); c) Fish treated with CLEO at concentrations of 125 \u0026micro;L.L⁻\u0026sup1;, 150 \u0026micro;L.L⁻\u0026sup1;, 175 \u0026micro;L.L⁻\u0026sup1;, 200 \u0026micro;L.L⁻\u0026sup1;, and 225 \u0026micro;L.L⁻\u0026sup1;. All fish were subjected to anesthetic induction for a duration of 5 minutes, followed by a 5-minute recovery period in water without CLEO. Each treatment group consisted of 9 fish, for a total of 108 juveniles of \u003cem\u003eColossoma macropomum\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e2.3.2 Experiment 1 - Behavioral Analysis of Anesthetic Induction and Recovery\u003c/h2\u003e \u003cp\u003eThe latency for the loss of posture reflex, characterized by lateral decubitus, was evaluated in fish exposed to CLEO concentrations of 125 \u0026micro;L.L⁻\u0026sup1;, 150 \u0026micro;L.L⁻\u0026sup1;, 175 \u0026micro;L.L⁻\u0026sup1;, 200 \u0026micro;L.L⁻\u0026sup1;, and 225 \u0026micro;L.L⁻\u0026sup1; (n\u0026thinsp;=\u0026thinsp;9 per treatment). After anesthetic induction, the latency for recovery of the posture reflex was assessed.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e \u003ch2\u003e2.3.3 Experiment 2 - Electrocardiogram (ECG) Analysis\u003c/h2\u003e \u003cp\u003eCardiac function was monitored in seven groups: a) Control group; b) Vehicle group; c) Groups treated with CLEO at concentrations of 125 \u0026micro;L.L⁻\u0026sup1;, 150 \u0026micro;L.L⁻\u0026sup1;, 175 \u0026micro;L.L⁻\u0026sup1;, 200 \u0026micro;L.L⁻\u0026sup1;, and 225 \u0026micro;L.L⁻\u0026sup1;, totaling 63 animals. Custom electrodes made from 0.3 mm diameter 925 silver with a length of 10 mm were used. The negative reference electrode was positioned at the base of the heart, and the positive recording electrode at the heart apex. The reference electrode was attached to the ventral portion of the left opercular opening (0.2 mm from the opercular cavity), and the recording electrode was inserted 2.0 mm from the right opercular opening. Electrodes were connected to a high-impedance amplifier (Grass Technologies, Model P511), enabling analysis of heart rate (bpm), QRS complex amplitude (mV), QRS complex duration (ms), R-R intervals (ms), P-Q intervals (ms), and Q-T intervals (ms).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e2.3.4 Recording and Analysis of ECG Data\u003c/h2\u003e \u003cp\u003eElectrodes were connected to a digital data acquisition system through a differential high-input impedance amplifier (Grass Technologies, Model P511), filtered at 0.3\u0026ndash;300 Hz, amplified 2000x, and monitored with an oscilloscope (ProteK, Model 6510). Signals were digitized at a rate of 1 kHz and stored on a hard drive for later processing using specialized software (LabVIEW Express). Data analysis was performed with a custom Python program utilizing Numpy and Scipy libraries for mathematical processing, and Matplotlib for graph generation. The graphical interface was developed using the PyQt4 library (Hamoy et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e \u003ch2\u003e2.3.5 Experiment 3 - Evaluation of Mechanism Underlying CLEO Effects\u003c/h2\u003e \u003cp\u003eTo assess the GABAergic allosteric activity of CLEO, fish were pretreated with flumazenil (0.1 mg/mL, 1 mg/kg i.p.) 15 minutes prior to CLEO treatment at concentrations of 125 \u0026micro;L.L⁻\u0026sup1;, 150 \u0026micro;L.L⁻\u0026sup1;, 175 \u0026micro;L.L⁻\u0026sup1;, 200 \u0026micro;L.L⁻\u0026sup1;, and 225 \u0026micro;L.L⁻\u0026sup1;. Anesthetic induction was observed until the loss of the posture reflex, followed by recovery time assessment.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Statistical Analysis\u003c/h2\u003e \u003cp\u003eData normality and homogeneity of variances were verified using the Kolmogorov-Smirnov and Levene tests, respectively. One-way ANOVA followed by Tukey's post-hoc test was performed to compare means. Statistical analysis was conducted using GraphPad Prism\u0026reg; 8, with significance levels set at *p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, **p\u0026thinsp;\u0026lt;\u0026thinsp;0.01, and ***p\u0026thinsp;\u0026lt;\u0026thinsp;0.001 for all comparisons.\u003c/p\u003e \u003c/div\u003e"},{"header":"3 Results","content":"\u003cp\u003eThe behavioral analysis revealed that CLEO induced a loss of posture reflex in fish, with higher doses resulting in shorter latencies for the onset of this reflex loss. Fish treated with 125 \u0026micro;L.L⁻\u0026sup1; of CLEO exhibited a mean latency for loss of posture reflex of 169.0\u0026thinsp;\u0026plusmn;\u0026thinsp;12.00 seconds, which was longer compared to the other groups. The group treated with 150 \u0026micro;L.L⁻\u0026sup1; had a mean latency of 147.6\u0026thinsp;\u0026plusmn;\u0026thinsp;11.90 seconds, while groups treated with 175 \u0026micro;L.L⁻\u0026sup1;, 200 \u0026micro;L.L⁻\u0026sup1;, and 225 \u0026micro;L.L⁻\u0026sup1; showed significantly shorter latencies of 129.1\u0026thinsp;\u0026plusmn;\u0026thinsp;11.99 seconds, 101.2\u0026thinsp;\u0026plusmn;\u0026thinsp;7.48 seconds, and 90.56\u0026thinsp;\u0026plusmn;\u0026thinsp;10.25 seconds, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eRecovery of the posture reflex in the group treated with 125 \u0026micro;L.L⁻\u0026sup1; of CLEO occurred in 123.9\u0026thinsp;\u0026plusmn;\u0026thinsp;13.30 seconds, which was shorter than the recovery times in the other groups: 150 \u0026micro;L.L⁻\u0026sup1; (168.9\u0026thinsp;\u0026plusmn;\u0026thinsp;11.67 seconds), 175 \u0026micro;L.L⁻\u0026sup1; (206\u0026thinsp;\u0026plusmn;\u0026thinsp;14.27 seconds), 200 \u0026micro;L.L⁻\u0026sup1; (245\u0026thinsp;\u0026plusmn;\u0026thinsp;13.04 seconds), and 225 \u0026micro;L.L⁻\u0026sup1; (275.4\u0026thinsp;\u0026plusmn;\u0026thinsp;16.98 seconds). All groups showed concentration-dependent recovery times, with higher concentrations resulting in longer recovery periods. This indicates that the anesthetic effect was reversible but occurred more slowly in fish exposed to higher concentrations (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe normal electrocardiogram of tambaqui exhibited a sinus rhythm, with a mean heart rate of 95.22\u0026thinsp;\u0026plusmn;\u0026thinsp;7.775 bpm. The P wave, QRS complex, and T wave were clearly identifiable (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA, B, and C). The R-R intervals (ms), P-Q intervals (ms), and Q-T intervals (ms) remained consistent with normal cardiac parameters, allowing for the assessment of treatment interference at increasing doses of CLEO. In the control group, the cardiac rhythm was sinusoidal throughout the 5-minute recording (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eCardiac activity in the control group exhibited a mean heart rate of 95.22\u0026thinsp;\u0026plusmn;\u0026thinsp;7.77 bpm, which was similar to the vehicle group (p\u0026thinsp;=\u0026thinsp;0.999), both maintaining a sinus rhythm with all cardiac deflections visible on the electrocardiogram (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA, B, C, and \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e A). The 30-second amplification allowed observation of all ECG elements: the P wave representing atrial contraction, the QRS complex indicating ventricular contraction, and the T wave representing ventricular repolarization. The 5-second amplification facilitated the evaluation of intervals during immersion treatment with CLEO and subsequent recovery (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eA, B, C, D, E, and F).\u003c/p\u003e \u003cp\u003eDuring immersion treatment with 125 \u0026micro;L.L⁻\u0026sup1;, fish exhibited a 15.28% decrease in heart rate compared to the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB). The group treated with 150 \u0026micro;L.L⁻\u0026sup1; of CLEO showed a 24.15% reduction (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eC), while the group treated with 175 \u0026micro;L.L⁻\u0026sup1; exhibited a 24.61% decrease (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eD). A 36.28% reduction was observed in the group treated with 200 \u0026micro;L.L⁻\u0026sup1; (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eE), and the group treated with 225 \u0026micro;L.L⁻\u0026sup1; presented a 45.85% decrease (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eF). Fish treated with CLEO displayed bradycardia while maintaining a sinus rhythm, with the intensity of bradycardia increasing alongside the concentration (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eB, C, D, E, and F).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe cardiac frequency decreased during treatment with increasing concentrations of CLEO. The control group had a mean frequency of 95.22\u0026thinsp;\u0026plusmn;\u0026thinsp;7.775 bpm, which was similar to the vehicle group (p\u0026thinsp;=\u0026thinsp;0.999), but higher than the treated groups. The group treated with 125 \u0026micro;L.L⁻\u0026sup1; had a mean frequency of 80.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.0 bpm, significantly higher than the other treated groups. The 150 \u0026micro;L.L⁻\u0026sup1; group (72.22\u0026thinsp;\u0026plusmn;\u0026thinsp;2.72 bpm) was similar to the 175 \u0026micro;L.L⁻\u0026sup1; group (p\u0026thinsp;=\u0026thinsp;0.999) but higher than the groups treated with 200 \u0026micro;L.L⁻\u0026sup1; and 225 \u0026micro;L.L⁻\u0026sup1;. The 225 \u0026micro;L.L⁻\u0026sup1; group had the lowest frequency compared to all other groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eThe mean amplitude of the QRS complex in the control group was 1.634\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23 mV, which was similar to the other groups (F(6, 56)\u0026thinsp;=\u0026thinsp;2.348; p\u0026thinsp;=\u0026thinsp;0.0430) (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eThe mean RR interval for the control group was 633.1\u0026thinsp;\u0026plusmn;\u0026thinsp;47.59 ms, similar to the vehicle group (p\u0026thinsp;=\u0026thinsp;0.999), but lower than the other treated groups. The group treated with 125 \u0026micro;L.L⁻\u0026sup1; had a mean RR interval of 743.9\u0026thinsp;\u0026plusmn;\u0026thinsp;28.50 ms, which was lower than the other treated groups. The 150 \u0026micro;L.L⁻\u0026sup1; group (831.4\u0026thinsp;\u0026plusmn;\u0026thinsp;30.49 ms) was similar to the 175 \u0026micro;L.L⁻\u0026sup1; group (p\u0026thinsp;=\u0026thinsp;0.999), while the 225 \u0026micro;L.L⁻\u0026sup1; group had a higher RR interval than the others (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003eThe mean PQ interval for the control group was 82.22\u0026thinsp;\u0026plusmn;\u0026thinsp;3.52 ms, with no significant difference from the vehicle group (p\u0026thinsp;=\u0026thinsp;0.999). The groups treated with 125 \u0026micro;L.L⁻\u0026sup1; (94.56\u0026thinsp;\u0026plusmn;\u0026thinsp;3.84 ms) and 150 \u0026micro;L.L⁻\u0026sup1; were similar (p\u0026thinsp;=\u0026thinsp;0.996). The 225 \u0026micro;L.L⁻\u0026sup1; group (140.0\u0026thinsp;\u0026plusmn;\u0026thinsp;4.52 ms) showed a greater PQ interval than the other groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eD).\u003c/p\u003e \u003cp\u003eThe mean duration of the QRS complex for the control group during induction was 19.33\u0026thinsp;\u0026plusmn;\u0026thinsp;1.80 ms, similar to the vehicle group and the group treated with 125 \u0026micro;L.L⁻\u0026sup1; (p\u0026thinsp;=\u0026thinsp;0.512), but shorter than the other groups. The 125 \u0026micro;L.L⁻\u0026sup1; group (20.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.30 ms) was similar to the 150 \u0026micro;L.L⁻\u0026sup1; group (p\u0026thinsp;=\u0026thinsp;0.7813). The group treated with 175 \u0026micro;L.L⁻\u0026sup1; was similar to the group treated with 200 \u0026micro;L.L⁻\u0026sup1; (p\u0026thinsp;=\u0026thinsp;0.2027). The 200 \u0026micro;L.L⁻\u0026sup1; group (28.44\u0026thinsp;\u0026plusmn;\u0026thinsp;1.81 ms) was similar to the 225 \u0026micro;L.L⁻\u0026sup1; group (p\u0026thinsp;=\u0026thinsp;0.512) (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eE).\u003c/p\u003e \u003cp\u003eFor the control group, the mean QT interval during induction was 211.9\u0026thinsp;\u0026plusmn;\u0026thinsp;14.94 ms, similar to the vehicle group (p\u0026thinsp;=\u0026thinsp;0.998). The group treated with 125 \u0026micro;L.L⁻\u0026sup1; (242.2\u0026thinsp;\u0026plusmn;\u0026thinsp;10.62 ms) was similar to the 150 \u0026micro;L.L⁻\u0026sup1; group (p\u0026thinsp;=\u0026thinsp;0.964). The 225 \u0026micro;L.L⁻\u0026sup1; group (338.4\u0026thinsp;\u0026plusmn;\u0026thinsp;10.35 ms) had a longer QT interval compared to the other groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003eF).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eDuring the recovery phase following exposure to CLEO at concentrations of 125 \u0026micro;L.L⁻\u0026sup1;, 150 \u0026micro;L.L⁻\u0026sup1;, 175 \u0026micro;L.L⁻\u0026sup1;, 200 \u0026micro;L.L⁻\u0026sup1;, and 225 \u0026micro;L.L⁻\u0026sup1;, reversibility of electrocardiographic alterations was observed (Figs.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA, B, C, D, and E). However, this reversibility was slower in groups treated with higher concentrations of CLEO.\u003c/p\u003e \u003cp\u003eDuring recovery, the group treated with 125 \u0026micro;L.L⁻\u0026sup1; showed results similar to the control group (p\u0026thinsp;=\u0026thinsp;0.4759) (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA). The fish treated with 150 \u0026micro;L.L⁻\u0026sup1; exhibited a 90.08% recovery of cardiac function compared to the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eB). For the group treated with 175 \u0026micro;L.L⁻\u0026sup1; of CLEO, the recovery was 86.34% (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eC), while the group treated with 200 \u0026micro;L.L⁻\u0026sup1; showed a recovery of 85.88% (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eD). Lastly, the group treated with 225 \u0026micro;L.L⁻\u0026sup1; also presented an 85.88% recovery (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eE). Although the treated fish exhibited a slow reversibility, they maintained a sinus rhythm (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eDuring recovery, the control group exhibited a mean heart rate of 95.22\u0026thinsp;\u0026plusmn;\u0026thinsp;7.77 bpm, similar to the vehicle group and the group treated with 125 \u0026micro;L.L⁻\u0026sup1; (p\u0026thinsp;=\u0026thinsp;0.4759). The groups treated with 150 \u0026micro;L.L⁻\u0026sup1; (85.11\u0026thinsp;\u0026plusmn;\u0026thinsp;2.26 bpm) were comparable to those treated with 175 \u0026micro;L.L⁻\u0026sup1;, 200 \u0026micro;L.L⁻\u0026sup1;, and 225 \u0026micro;L.L⁻\u0026sup1; (p\u0026thinsp;=\u0026thinsp;0.5156) (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eThe amplitude of the QRS complex during recovery for the control group was 1.634\u0026thinsp;\u0026plusmn;\u0026thinsp;0.238 mV, similar to the other groups (F(6, 56)\u0026thinsp;=\u0026thinsp;0.5940, p\u0026thinsp;=\u0026thinsp;0.7338) (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eThe mean RR interval during recovery for the control group was 633.1\u0026thinsp;\u0026plusmn;\u0026thinsp;47.59 ms, comparable to the vehicle and 125 \u0026micro;L.L⁻\u0026sup1; groups (p\u0026thinsp;=\u0026thinsp;0.5850). The groups treated with 150 \u0026micro;L.L⁻\u0026sup1; (704.9\u0026thinsp;\u0026plusmn;\u0026thinsp;19.63 ms) were similar to the 175 \u0026micro;L.L⁻\u0026sup1;, 200 \u0026micro;L.L⁻\u0026sup1;, and 225 \u0026micro;L.L⁻\u0026sup1; groups (p\u0026thinsp;=\u0026thinsp;0.2349) (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003eThe PQ interval during recovery for the control was 82.22\u0026thinsp;\u0026plusmn;\u0026thinsp;3.528 ms, similar to the vehicle and treated groups with 125 \u0026micro;L.L⁻\u0026sup1;, 150 \u0026micro;L.L⁻\u0026sup1;, and 175 \u0026micro;L.L⁻\u0026sup1; (p\u0026thinsp;=\u0026thinsp;0.0914). The groups treated with 200 \u0026micro;L.L⁻\u0026sup1; and 225 \u0026micro;L.L⁻\u0026sup1; showed higher means than the control and vehicle groups yet were similar to the other treated groups (p\u0026thinsp;=\u0026thinsp;0.091) (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eD).\u003c/p\u003e \u003cp\u003eThe duration of the QRS complex during recovery for the control group was 19.33\u0026thinsp;\u0026plusmn;\u0026thinsp;1.80 ms, similar to the vehicle and groups treated with 125 \u0026micro;L.L⁻\u0026sup1;, 150 \u0026micro;L.L⁻\u0026sup1;, 175 \u0026micro;L.L⁻\u0026sup1;, and 200 \u0026micro;L.L⁻\u0026sup1; (p\u0026thinsp;=\u0026thinsp;0.3891). The groups treated with 225 \u0026micro;L.L⁻\u0026sup1; were similar to the groups treated with 175 \u0026micro;L.L⁻\u0026sup1; and 200 \u0026micro;L.L⁻\u0026sup1; (p\u0026thinsp;=\u0026thinsp;0.3891) (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003eE).\u003c/p\u003e \u003cp\u003eDuring recovery, the QT interval for the control was 211.9\u0026thinsp;\u0026plusmn;\u0026thinsp;14.94 ms, similar to the vehicle, 125 \u0026micro;L.L⁻\u0026sup1;, and 150 \u0026micro;L.L⁻\u0026sup1; groups (p\u0026thinsp;=\u0026thinsp;0.9957). The groups treated with 175 \u0026micro;L.L⁻\u0026sup1; (228.7\u0026thinsp;\u0026plusmn;\u0026thinsp;4.92 ms) were similar to the 200 \u0026micro;L.L⁻\u0026sup1; and 225 \u0026micro;L.L⁻\u0026sup1; groups (p\u0026thinsp;=\u0026thinsp;0.7295) but were higher than the other groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAfter the administration of flumazenil (1 mg/kg, i.p.), there was an increase in latency for the loss of postural reflex behavior across all tested concentrations of CLEO. The group treated with flumazenil followed by immersion in 125 \u0026micro;L.L⁻\u0026sup1; showed an increase of 263.13% in latency. Similar increases were observed in the groups treated with 150 \u0026micro;L.L⁻\u0026sup1; (208.02%), 175 \u0026micro;L.L⁻\u0026sup1; (181.4%), 200 \u0026micro;L.L⁻\u0026sup1; (223.22%), and 225 \u0026micro;L.L⁻\u0026sup1; (215.77%), indicating that after flumazenil administration, latencies were significantly prolonged (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eDuring the recovery period, the group that received flumazenil exhibited shorter latencies for recovering the postural reflex after treatment with different concentrations of CLEO. The group treated with 125 \u0026micro;L.L⁻\u0026sup1; showed a decrease in latency for recovery of the postural reflex by 53.99%. Similar reductions were noted in the groups treated with 150 \u0026micro;L.L⁻\u0026sup1; (48.18%), 175 \u0026micro;L.L⁻\u0026sup1; (52.10%), 200 \u0026micro;L.L⁻\u0026sup1; (43.26%), and 225 \u0026micro;L.L⁻\u0026sup1; (38.99%) (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e8\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"4 Discussion","content":"\u003cp\u003eThe results from the chromatogram in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, obtained via gas chromatography, revealed the predominance of three terpenes in the essential oil, with Ar-turmerone as the major component (32.5%), followed by Curcumene (13%) and Farnesene (7.2%). The presence of Ar-turmerone as the primary component has been confirmed by other studies (Zhang et al., \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Guimar\u0026atilde;es et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Zheng et al., \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), and its therapeutic potential is widely recognized (Avan\u0026ccedil;o et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Kheira et al., 2020). The anesthetic properties of Ar-turmerone are currently being investigated, given its chemical similarity to synthetic anesthetics, such as propofol, commonly used in fish anesthesia.\u003c/p\u003e \u003cp\u003eBehavioral parameters observed during both induction and recovery from anesthesia showed a dose-dependent effect, with groups receiving higher doses exhibiting a more rapid loss of postural reflex and a slower reversibility (Dos Santos et al., 2022; Vieira et al., \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Dos Santos et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). The anesthetic induction induced by turmeric oil can be explained by its terpene-rich composition, particularly the Ar-turmerone, which, due to its structure, can modulate GABA A receptors (Bianchini et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Kasai et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Khumpirapang et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This mechanism is similar to that of anesthetics used in humans (Zhou et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eOur findings suggest that lower doses of the oil are safer for use, as evidenced by the group exposed to the dose of 125 \u0026micro;L.L⁻\u0026sup1;, which showed a recovery of 90.08% in cardiac function, similar to the control group. This contrasts with the study by Saccol et al. (\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), which used significantly higher doses (200 \u0026micro;L.L⁻\u0026sup1;, 300 \u0026micro;L.L⁻\u0026sup1;, and 500 \u0026micro;L.L⁻\u0026sup1;) and observed anesthesia only in groups exposed to 200 \u0026micro;L.L⁻\u0026sup1;.\u003c/p\u003e \u003cp\u003eA reduction in heart rate (HR) was observed in fish exposed to turmeric oil compared to the control group, with a direct relationship between concentration and cardiac depression. These results are consistent with studies showing that menthol administration also reduced cardiac activity in Colossoma macropomum in a dose-dependent manner, with reversibility to baseline levels (Cantanh\u0026ecirc;de et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Moreover, several studies have reported a reduction in the cardiac function of Tambaqui when exposed to different anesthetic concentrations derived from essential oils, with complete recovery post-anesthesia (De Souza et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Barbas et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe prolongation of QT and RR intervals observed in Colossoma macropomum exposed to turmeric oil, compared to the control group, indicates cardiac alterations related to increased oil concentration. However, these effects did not result in impaired cardiac function, aligning with findings from Cantanh\u0026ecirc;de et al. (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) and Da Costa et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The absence of atrioventricular block (AVB) suggests cardiovascular safety, as noted by Zena et al. (\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe analysis of the QRS interval showed an increase in its duration with the increasing dose of turmeric oil, but the anesthetic induction with the minimum dose was similar to the control group, indicating that cardiac contraction was maintained. These findings are corroborated by studies by De Sousa et al. (2019) and Vilhena et al. (\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eRegarding the evaluation of CLEO in relation to the GABA A-benzodiazepine receptor, it appears that the components of the oil act synergistically with the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). The pre-treatment with flumazenil, a benzodiazepine receptor antagonist, increased the latency for the loss of postural reflex and accelerated recovery, confirming the involvement of GABA A receptors in the anesthesia induced by CLEO.\u003c/p\u003e \u003cp\u003eThis study demonstrated the anesthetic activity of CLEO at various treatment concentrations, evidencing the reversibility of behavioral and cardiac effects. According to our results, the safe range for short-duration anesthesia appears to be between 125 and 225 \u0026micro;L.L⁻\u0026sup1;, while for deeper anesthesia, we recommend concentrations of 200 to 225 \u0026micro;L.L⁻\u0026sup1;, without significant impairments in cardiac activity. The mechanism of action of CLEO components involves the potentiation of the GABAergic system, as indicated by the action of flumazenil, suggesting that the anesthesia induced by CLEO is mediated by allosteric mechanisms at the GABA A receptors.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eConflict of interest\u003c/h2\u003e \u003cp\u003eThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.\u003c/p\u003e \u003ch2\u003ePublisher\u0026rsquo;s note\u003c/h2\u003e \u003cp\u003eAll claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.\u003c/p\u003e \u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThe author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by Funda\u0026ccedil;\u0026atilde;o Amaz\u0026ocirc;nia de Amparo a Estudos e Pesquisas do Estado do Par\u0026aacute; (FAPESPA) and The APC was funded by Pr\u0026oacute;-Reitoria de Pesquisa e P\u0026oacute;s Gradua\u0026ccedil;\u0026atilde;o\u0026mdash;PROPESP/UFPA.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eConceived and designed the experiments: L.E.C and M.H. Performed the experiments: L.E.C., L.V.d.S., L.L.d.R., G.B.B and M.H. Writing-original draft and editing: all authors. Financial support and administrative support: M.H. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e \u003cp\u003eThe authors would like to acknowledge Funda\u0026ccedil;\u0026atilde;o Amaz\u0026ocirc;nia de Amparo a Estudos e Pesquisas do Estado do Par\u0026aacute; (FAPESPA) for their financial support provided of this research, Programa de P\u0026oacute;s gradua\u0026ccedil;\u0026atilde;o em Ci\u0026ecirc;ncias Farmac\u0026ecirc;uticas (PPGCF) and Programa de P\u0026oacute;s gradua\u0026ccedil;\u0026atilde;o em Farmacologia e Bioqu\u0026iacute;mica (PPGFARMABIO).\u003c/p\u003e\u003ch2\u003eData availability statement\u003c/h2\u003e \u003cp\u003eThe original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding authors.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbdel-Aziz MFA (2013) Effect of some environmental conditions and protein quality on reproductive and productive performance of red hybrid tilapia (Oreochromis sp.). MSc thesis, Animal Dep., Fac. Agri., Fayoum university, Egypt.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAggarwal B, Harikumar K (2009) Potential therapeutic effects of curcumin, the antiinflammatory agent, against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplastic diseases. Int. J. Biochem. Cell Biol, 41(1), 40\u0026ndash;59. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.biocel.2008.06.010\u003c/span\u003e\u003cspan address=\"10.1016/j.biocel.2008.06.010\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eArnaout R, Ferrer T, Huisken J, Spitzer K, Stainier DYR, Tristani-Firouzi M, et al (2007) \u0026ldquo;Zebrafish model for human long QT syndrome,\u0026rdquo; Proceedings of the National Academy of Sciences, vol. 104, p. 11316. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1073/pnas.0702724104\u003c/span\u003e\u003cspan address=\"10.1073/pnas.0702724104\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAvan\u0026ccedil;o GB, Ferreira FD, Bomfim NS, de Souza Rodrigues dos Santos PA, Peralta RM, Brugnari T, Mallmann CA, de Abreu Filho BA, Mikcha JMG, Machinski M (2017) Curcuma longa L. essential oil composition, antioxidant effect, and effect on Fusarium verticillioides and fumonisin production. Food Control 73:806\u0026ndash;813. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.foodcont.2016.09.032\u003c/span\u003e\u003cspan address=\"10.1016/j.foodcont.2016.09.032\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAydın I, Akbulut B, Kucuk, E, Kumlu M (2015) Effects of temperature, fish size and dosage of clove oil on anaesthesia in turbot (Psetta maxima Linnaeus). Journal of Fisheries and Aquatic Science. Sci, 15, 899\u0026ndash;904. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4194/1303-2712-v15_4_13\u003c/span\u003e\u003cspan address=\"10.4194/1303-2712-v15_4_13\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBarbas LAL, Hamoy M, de Mello VJ, Barbosa RPM, de Lima HST, Torres MF, Gomes, MRF (2017) Essential oil of citronella modulates electrophysiological responses in tambaqui Colossoma macropomum: a new anaesthetic for use in fish. Aquaculture, 479, 60\u0026ndash;68. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aquaculture.2017.05.027\u003c/span\u003e\u003cspan address=\"10.1016/j.aquaculture.2017.05.027\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBarbas LAL, Maltez LC, Stringhetta GR, de Oliveira GL, Monserrat JM, da Silva DT, Sampaio LA (2017) Properties of two plant extractives as anaesthetics and antioxidants for juvenile tambaqui Colossoma macropomum. Aquaculture, 469, 79\u0026ndash;87. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aquaculture.2016.12.012\u003c/span\u003e\u003cspan address=\"10.1016/j.aquaculture.2016.12.012\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBarbas LAL, Stringhetta GR, de Oliveira GL, Figueiredo MRC, Sampaio LA (2016) Jambu, Spilanthes acmella as a novel anaesthetic for juvenile tambaqui, Colossoma macropomum: secondary stress responses during recovery. Aquaculture, 456, 70\u0026ndash;75. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aquaculture.2016.01.026\u003c/span\u003e\u003cspan address=\"10.1016/j.aquaculture.2016.01.026\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBecker AG, Parodi TV, Heldwein CG, Zeppenfeld CC, Heinzmann BM, Baldisserotto B (2012) Transportation of silver catfish, Rhamdia quelen, in water with eugenol and the essential oil of Lippia alba. Fish Physiol. Biochem. 38, 789\u0026ndash;796. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s10695-011-9562-4\u003c/span\u003e\u003cspan address=\"10.1007/s10695-011-9562-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBianchini AE, Garlet QI, Da Cunha JA, Bandeira Junior G, Brusque ICM, Salbego J, Heinzmann BM, Baldisserotto B (2017) Monoterpenoids (Thymol, carvacrol and S-(+)- linalool) with anesthetic activity in silver catfish (Rhamdia quelen): Evaluation of acetylcholinesterase and GABaergic activity. Brazilian J. Med. Biol. Res. 50, 1\u0026ndash;8. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1590/1414-431x20176346\u003c/span\u003e\u003cspan address=\"10.1590/1414-431x20176346\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCantanh\u0026ecirc;de SM, Amado LL, da Costa BMPA, Barbas LAL, Torres MF, Hamoy AO, Hamoy M (2021) Menthol exposure induces reversible cardiac depression and reduces lipid peroxidation in the heart tissue of tambaqui Colossoma macropomum. Aquaculture, 541, 736847. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aquaculture.2021.736847\u003c/span\u003e\u003cspan address=\"10.1016/j.aquaculture.2021.736847\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCao Y, Yan H, Yu G, Su R (2019) Flumazenil-insensitive benzodiazepine binding sites in GABA\u003csub\u003eA\u003c/sub\u003e receptors contribute to benzodiazepine-induced immobility in zebrafish larvae. Life Sci. 15;239:117033. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.lfs.2019.117033\u003c/span\u003e\u003cspan address=\"10.1016/j.lfs.2019.117033\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eda Costa, BMA, Torres, MF, da Silva RA, Aydın B, Amado LL, Hamoy M, Barbas LAL (2022) Integrated behavioural, neurological, muscular and cardiorespiratory response in tambaqui, Colossoma macropomum anaesthetized with menthol. Aquaculture, 560, 738553. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aquaculture.2022.738553\u003c/span\u003e\u003cspan address=\"10.1016/j.aquaculture.2022.738553\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDavis MW (2010) Fish stress and mortality can be predicted using reflex impairment. Fish and Fisheries, 11(1), 1\u0026ndash;11. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1467-2979.2009.00331.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1467-2979.2009.00331.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ede Souza, ADSL, Peret AC, Hamoy M, de Souza RAL, Torres MF, Barbas LAL (2019) Propofol and essential oil of Nepeta cataria induce anaesthesia and marked myorelaxation in tambaqui Colossoma macropomum: implications on cardiorespiratory responses. Aquaculture, 500,160\u0026ndash;169. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aquaculture.2018.10.017\u003c/span\u003e\u003cspan address=\"10.1016/j.aquaculture.2018.10.017\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDelbon MC, Paiva MR (2012) Eugenol in tilapia juvenile: concentrations and successive administrations. Boletim do Instituto de Pesca, 38(1): 43\u0026ndash;52.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDos Santos MF, do Nascimento LM, da Paz CA, C\u0026acirc;mara TM, Motomya YKM, da Cunha Ferreira R, da Silva Deiga Y, Monteiro E, Cantanh\u0026ecirc;de SM, Amado LL, Hamoy M (2024) Behavioral and electrophysiological study in Colossoma macropomum treated with different concentrations of Nepeta cataria oil in an immersion bath revealed a therapeutic window for anesthesia. Fish Physiol Biochem. Aug;50(4):1651\u0026ndash;1665. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s10695-024-01361-2\u003c/span\u003e\u003cspan address=\"10.1007/s10695-024-01361-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFaleschini T, Syafni N, Schulte HL, Garifulina A, Hering S, Espindola LS, Hamburger M (2023) A neolignan from Connarus tuberosus as an allosteric GABA\u003csub\u003eA\u003c/sub\u003e receptor modulator at the neurosteroid binding site. Biomed Pharmacother. 2023;161:114498. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.biopha.2023.114498\u003c/span\u003e\u003cspan address=\"10.1016/j.biopha.2023.114498\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFAO (2014) Food and Agriculture Organization of the United Nations.El estado mundial de la pesca y la acuicultura. Oportunidades y desaf\u0026iacute;os. 274p.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFelix LM, Luzio A, Antunes L, Coimbra AM, Valentim AM (2021) Malformations and mortality in zebrafish early stages associated with elevated caspase activity after 24 h exposure to MS-222. Toxicology and Applied Pharmacology, 412, Article 115385. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.taap.2020.115385\u003c/span\u003e\u003cspan address=\"10.1016/j.taap.2020.115385\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGenge CE, Lin E, Lee L, Sheng X, Rayani K, Gunawan M, Stevens CM, Li AY, Talab SS, Claydon TW, Hove-Madsen L, Tibbits GF (2016) Reviews of Physiology, Biochemistry and Pharmacology, vol. 171. Springer International Publishing, Cham, pp. 99\u0026ndash;136.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGoulding M, Carvalho M (1982) Life history and management of the tambaqui (Colossoma macropomum, Characidae): an important Amazonian food fish. Revista Brasileira de Zoologia, 1, 107\u0026ndash;133. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1590/S0101-81751982000200001\u003c/span\u003e\u003cspan address=\"10.1590/S0101-81751982000200001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuilherme HO, Santos FAC, Costa LS, Prado VGL, Palheta GDA, de Melo NFAC, Luz RK, Ribeiro PAP (2022) Feeding, growth, and blood chemistry of the tambaqui (Colossoma macropomum) held under self-feeding and time-restricted automatic feeding conditions. Aquac. Int. 30, 323\u0026ndash;339.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10499-021-00801-7\u003c/span\u003e\u003cspan address=\"10.1007/s10499-021-00801-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuimar\u0026atilde;es IG, Miranda EC, Ara\u0026uacute;jo JG (2014) Coefficients of total tract apparent digestibility of some feedstuffs for Tambaqui (Colossoma macropomum). Animal Feed Science and Technology, 188, 150\u0026ndash;155. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.anifeedsci.2013.11.007\u003c/span\u003e\u003cspan address=\"10.1016/j.anifeedsci.2013.11.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuimar\u0026atilde;es AFAC, Vinhas AF, Gomes LH, Souza PBK (2020) Essential oil of Curcuma longa Rhizomes chemical composition, yield variation stability. Quim. Nova, Vol. 43, No. 7, 909\u0026ndash;913. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.21577/0100-4042.20170547\u003c/span\u003e\u003cspan address=\"10.21577/0100-4042.20170547\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHamoy AO, Fonseca SMD, Cei GL, J\u0026uacute;nior FLDA, Hamoy MKO, Ribeiro RM, Barbas LAL, Muto N, Hamoy M (2023) Behavioral, electrocorticographic and electrocardiologic changes in Colossoma macropomum (Tambaqui) in the effect of cunaniol. PLoS One. Jun 30;18(6):e0287681. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal.pone.0287681\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0287681\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHarmon TS, (2009) Methods for reducing stressors and maintaining water quality associated with live fish transport in tanks: a review of the basics. Reviews in Aquaculture, 1, 58\u0026ndash;66. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1753-5131.2008.01003.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1753-5131.2008.01003.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHasan H, Farida EG (2016) Different concentration of rubber seed extract (Hevea brasiliensi) for anesthetic to the survival of prospective broodstock milkfish by using closed transportation. Jurnal Ruaya, 4(2),55\u0026ndash;62.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aaf.201903002\u003c/span\u003e\u003cspan address=\"10.1016/j.aaf.201903002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHohlenwerger JC, Baldisserotto B, Couto RD, Heinzmann BM, da Silva DT, Caron BO, Schmidt D, Copatti CE (2017) Essential oil of Lippia alba in the transport of Nile tilapia. Ci\u0026ecirc;ncia Rural, 47(3). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1590/0103-8478cr20160040\u003c/span\u003e\u003cspan address=\"10.1590/0103-8478cr20160040\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHoseini SM, Mirghaed AT, Yousefi M (2019) Application of herbal anaesthetics in aquaculture. Reviews in Aquaculture, 11, 550\u0026ndash;564. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/raq.12245\u003c/span\u003e\u003cspan address=\"10.1111/raq.12245\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIBGE (2020) Instituto Brasileiro de Geografia e Estat\u0026iacute;stica \u0026ndash; Produ\u0026ccedil;\u0026atilde;o de aquicultura.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.sidra.ibge.gov.br/bda/tabela/listabl.asp?c=3940\u0026amp;z=t\u0026amp;o=21\u003c/span\u003e\u003cspan address=\"http://www.sidra.ibge.gov.br/bda/tabela/listabl.asp?c=3940\u0026amp;z=t\u0026amp;o=21\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. (Accessed 19.09.2024).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKasai M, Hososhima S, Yun- Fei L (2014) Menthol induces surgical anesthesia and rapid movement in fishes. The Open Neuroscience Journal, 8, 1\u0026ndash;8. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2174/1874082001408010001\u003c/span\u003e\u003cspan address=\"10.2174/1874082001408010001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKheira M, Boumediene M, Aicha TT, Pascal S (2023) Chemical Composition and Antioxidant and Antimicrobial Activities of Curcuma longa L. Essential Oils. Egyptian Journal of Chemistry 66(7): 283\u0026ndash;295. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.21608/ejchem.2022.121586.5555\u003c/span\u003e\u003cspan address=\"10.21608/ejchem.2022.121586.5555\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKhumpirapang N, Chaichit S, Jiranusornkul S, Pikulkaew S, Mullertz A, Okonogi S (2018) In vivo anesthetic effect and mecha nism of action of active compounds from Alpinia galanga oil on Cyprinus carpio (koi carp). Aquaculture, 496, 176\u0026ndash;184. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aquaculture.2018.07.017\u003c/span\u003e\u003cspan address=\"10.1016/j.aquaculture.2018.07.017\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKim HT, Lee JH, Park IB, Heo HE, Kim TY, Lee MJ (2010) Long QT syndrome provoked by induction of general anesthesia-A case report. Korean Journal of Anesthesiology, 59(Suppl), S114-S118. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.4097/kjae.2010.59.S.S114\u003c/span\u003e\u003cspan address=\"10.4097/kjae.2010.59.S.S114\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKoopman CD, De Angelis J, Iyer SP, Verkerk AO, Da Silva J, Berecki G, Jeanes A, Baillie GJ, Paterson S, Uribe V, Ehrlich OV, Robinson SD, Garric L, Petrou S, Simons C, Vetter I, Hogan BM, de Boer TP, Bakkers J, Smith KA (2021) The zebrafish grime mutant uncovers an evolutionarily conserved role for Tmem161b in the control of cardiac rhythm. Proc Natl Acad Sci USA.;11:8. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1073/pnas.2018220118\u003c/span\u003e\u003cspan address=\"10.1073/pnas.2018220118\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLe T, Zhang J, Nguyen AH, Trigo Torres RS, Vo K, Dutt N, Lee J, Ding Y, Xu X, Lau MPH, Cao H (2022) A novel wireless ECG system for prolonged monitoring of multiple zebrafish for heart disease and drug screening studies. Biosens Bioelectron. Feb 1;197:113808. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bios.2021.113808\u003c/span\u003e\u003cspan address=\"10.1016/j.bios.2021.113808\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLuz JGR, Nogueira JN, Alves CMG, Videira MN, Canuto KM, Castro KNC, Tavares-Dias M (2021) Essential oil of Alpinia zerumbet (Zingiberaceae) has anthelmintic efficacy against monogenean of Colossoma macropomum (Characiformes: Serrasalmidae). Aquac. Res. 52, 5340\u0026ndash;5349. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/are.15404\u003c/span\u003e\u003cspan address=\"10.1111/are.15404\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMcGrath M, Hoyt H, Pence A, Forman SA, Raines DE (2021) Selective actions of benzodiazepines at the transmembrane anaesthetic binding sites of the GABA\u003csub\u003eA\u003c/sub\u003e receptor: In vitro and in vivo studies. Br J Pharmacol. Dec;178(24):4842\u0026ndash;4858. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/bph.15662\u003c/span\u003e\u003cspan address=\"10.1111/bph.15662\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMerola N, Cantelmo AO (1987) Growth, feed conversion and mortality of cagereared tambaqui, Colossoma macropomum, fed various dietary feeding regimes and protein levels. Aquaculture, 66(3\u0026ndash;4), 223\u0026ndash;233. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/0044-8486(87)90108-6\u003c/span\u003e\u003cspan address=\"10.1016/0044-8486(87)90108-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e;\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNakayama CL, Silva LFS, dos Santos FAC, Boaventura TP, Favero GC, Palheta GDA, Melo NFAC, Romano LA, Luz RK (2022) Zootechnical performance and some physiological indices of tambaqui, Colossoma macropomum juveniles during biofloc maturation and in different feed regimes. Agriculture 12, 1025. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agriculture12071025\u003c/span\u003e\u003cspan address=\"10.3390/agriculture12071025\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eParveen Z, Nawaz S, Siddique S, Shahzad K (2013) Composition and antimicrobial activity of the essential oil from leaves of Curcuma longa L. Kasur Variety. Indian Journal of Pharmaceutical Sciences, 75(1), 117\u0026ndash;122. https://dx.doi.org/10.4103%2F0250-474X.113544\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePEIXE BR (2024). Anu\u0026aacute;rio Peixe-BR da piscicultura 2024. S\u0026atilde;o Paulo.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePriborsky J, Velisek J (2018) A review of three commonly used fish anesthetics. Rev. Fish. Sci. Aquac. 26, 417\u0026ndash;442. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/23308249.2018\u003c/span\u003e\u003cspan address=\"10.1080/23308249.2018\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. 1442812\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePrimadona R, Lestari S, Baehaki A (2017) The effect of electric current on survival rate of fry finger (Clarias sp.) in dry transportation. Fistech-Jurnal Teknologi Hasil Perikanan, 6(2): 145\u0026ndash;152. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.36706/fishtech.v6i2.5846\u003c/span\u003e\u003cspan address=\"10.36706/fishtech.v6i2.5846\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePriya R, Prathapan A, Raghu KG, NirmalaMenon A (2012) Chemical composition and in vitro antioxidative potential of essential oil isolated from Curcuma longa L. leaves. Asian Pac J Trop Biomed. S695\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRamanayaka JC, Atapattu NSBM (2006) Fish Anaesthetic properties of some local plant material. Tropical Agricultural Research and Extension, 9,1\u0026ndash;6. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4038/tare.v9i0.5413\u003c/span\u003e\u003cspan address=\"10.4038/tare.v9i0.5413\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRezende FP, Pascoal LM, Vianna RA, Lanna EAT (2017) Sedation of nile tilapia with essential oils: Tea tree, clove, Eucalyptus, and mint oils. Rev. Caatinga, Mossor\u0026oacute;. 30(2), 479\u0026ndash;486. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1590/1983-21252017v30n223rc\u003c/span\u003e\u003cspan address=\"10.1590/1983-21252017v30n223rc\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRibeiro AS, Batista ES, Dairiki JK, Chaves FCM, Inoue LA (2016) Anesthetic properties of Ocimum gratissimum essential oil for juvenile matrinx\u0026atilde;. Acta Sci. Anim. Sci. 38, 1\u0026ndash;7. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.4025/actascianimsci.v38i1.28787\u003c/span\u003e\u003cspan address=\"10.4025/actascianimsci.v38i1.28787\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaccol EMH, Toni C, Pess TS, Ourique GM, Gressler LT, Silva LVF, Mour\u0026atilde;o RHV, Oliveira RB, Baldisserotto B, Pavanato MA (2017) Anesthetic and antioxidant effects of Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils oils on tambaqui (Colossoma macropomum). Aquac. Res. 48, 2012\u0026ndash;2031. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/are.13034\u003c/span\u003e\u003cspan address=\"10.1111/are.13034\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSchramm A, Ebrahimi SN, Raith M, Zaugg J, Rueda DC, Hering S, Hamburger M (2013) Phytochemi- cal profiling of Curcuma kwangsiensis rhizome extract, and identification of labdane diterpenoids as positive GABAA receptor modulators. Phytochemistry 96, 318\u0026ndash;329. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.phytochem.2013.08.004\u003c/span\u003e\u003cspan address=\"10.1016/j.phytochem.2013.08.004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSuwandi R, Nugraha R, Zulfamy KE (2013) Aplikasi ekstrak daun jambu Psidium guajava var.pomifera pada proses transportasi ikan nila. Oreochromis niloticus) JPHPI,16(1), 69\u0026ndash;78. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.17844/jphpi.v16i1.8107\u003c/span\u003e\u003cspan address=\"10.17844/jphpi.v16i1.8107\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTeixeira RR, de Souza RC, Sena AC, Baldisserotto B, Heinzmann BM, Copatti CE (2018) Essential oil of Aloysia Triphylla is effective in Nile Tilapia transport. Bol. Inst. Pesca 44, 17\u0026ndash;24. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.20950/1678-2305.2018.263\u003c/span\u003e\u003cspan address=\"10.20950/1678-2305.2018.263\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVieira LR, Pereira YLG, Diniz LA, Nascimento CP, Da Silva ALM, Azevedo JEC, De Mello VJ, Muto NA, Barbas LAL, Hamoy M (2023) Graded concentrations of lidocaine hydrochloride in the modulation of behavioral, cardiac, and muscular responses of the Amazon freshwater fsh tambaqui (Colossoma macropomum). Aquaculture. 563:738985. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.aquaculture.2022.738985\u003c/span\u003e\u003cspan address=\"10.1016/j.aquaculture.2022.738985\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVieira ELP, Egger RC, Lima TC, Paiva MB, Gon\u0026ccedil;alves LU, Honczaryk A, Godoy L, Tavares GC, Kuradomi RY (2022) Short-term preservation of Colossoma macropomum oocytes by different methods and chilling temperatures. Theriogenology 191, 22\u0026ndash;34. \u003cdiv class=\"ExternalRefDOI\"\u003ehttps://doi.org/10.1016/j\u003c/div\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVijayasteltar BL, Kottarapat J, Ramadasan K (2011) An evaluation of antioxidant, anti-inflammatory, and antinociceptive activities of essential oil from Curcuma longa L. Indian Journal of pharmaceutical Sciences, 43(5), 526\u0026ndash;531. https://dx.doi.org/10.4103%2F0253-7613.84961\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVilhena CS, da Silva RA, da Costa BMPA, Torres MF, de Mello VJ, Noronha RCR, do Ros\u0026aacute;rio da Silva JK, Hamoy M, Barbas LAL, do Nascimento LAS (2022) Cardiac response in tambaqui Colossoma macropomum anaesthetised with Piper divaricatum essential oil. Fish Physiol Biochem. 48(5):1413\u0026ndash;1425. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s10695-022-01132-x\u003c/span\u003e\u003cspan address=\"10.1007/s10695-022-01132-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWolkers CP, Barbosa Junior A, Menescal-de-Oliveira L, Hoffmann A (2015) GABAA-benzodiazepine receptors in the dorsomedial (Dm) telencephalon modulate restraint-induced antinociception in the fish Leporinus macrocephalus. Physiol Behav.Aug 1;147:175\u0026thinsp;\u0026ndash;\u0026thinsp;82. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.physbeh.2015.04.037\u003c/span\u003e\u003cspan address=\"10.1016/j.physbeh.2015.04.037\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYu ZF, Kong LD, Chen Y (2002) Antidepressant activity of aqueous extracts of Curcuma longa in mice. Journal of Ethnopharmacology 83, 161\u0026ndash;165. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/s0378-8741(02)00211-8\u003c/span\u003e\u003cspan address=\"10.1016/s0378-8741(02)00211-8\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZahl IH, Samuelsen O, Kiessling A (2012) Anaesthesia of farmed fish: implications for welfare. Fish Phys- iol Biochem 38:201\u0026ndash;218. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10695-011-9565-1\u003c/span\u003e\u003cspan address=\"10.1007/s10695-011-9565-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZena LA, Ekstr\u0026ouml;m A, Gr\u0026auml;ns A, Olsson C, Axelsson M, Sundh H, Sandblom E (2021) It takes time to heal a broken heart: ventricular plasticity improves heart performance after myocardial infarction in rainbow trout, Oncorhynchus mykiss. J Exp Biol. Dec 1;224(23):jeb243578. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1242/jeb.243578\u003c/span\u003e\u003cspan address=\"10.1242/jeb.243578\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang L, Yang Z, Chen F, Su P, Chen D, Pan W (2017) Composition and bioactivity assessment of essential oils of Curcuma longa L. collected in China. Ind Crops Prod.;109(15):60\u0026ndash;73.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZheng Y, Pan C, Zhang Z, Luo W, Liang X, Shi Y, Liang L, Zheng X, Zhang L, Du Z (2020) Antiaging effect of Curcuma longa L. essential oil on ultravioletirradiated skin. Microchemical Journal 154.104608. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.microc.2020.104608\u003c/span\u003e\u003cspan address=\"10.1016/j.microc.2020.104608\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhou C, Liu J, Chen XD (2012) General anesthesia mediated by effects on ion channels. World Journal of Critical Care Medicine, 1, 80\u0026ndash;93. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.5492/wjccm.v1.i3.80\u003c/span\u003e\u003cspan address=\"10.5492/wjccm.v1.i3.80\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Curcuma longa, electrocardiographic, behavioral, Colossoma macropomum, tambaqui","lastPublishedDoi":"10.21203/rs.3.rs-5154092/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5154092/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe use of natural agents as anesthetics in fish management aims to minimize stress during procedures. This study aims to investigate the behavioral, electrocardiographic characteristics, and the pharmacological mechanism of action of \u003cem\u003eCurcuma longa\u003c/em\u003e essential oil (CLEO) in \u003cem\u003eColossoma macropomum\u003c/em\u003e. The study was conducted on juvenile \u003cem\u003eColossoma macropomum\u003c/em\u003e (18.13\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1g) (n\u0026thinsp;=\u0026thinsp;153), using CLEO concentrations of 125 \u0026micro;L.L⁻\u0026sup1;, 150 \u0026micro;L.L⁻\u0026sup1;, 175 \u0026micro;L.L⁻\u0026sup1;, 200 \u0026micro;L.L⁻\u0026sup1;, and 250 \u0026micro;L.L⁻\u0026sup1;, analyzing anesthetic induction and recovery behavior (Experiment I), electrocardiogram (Experiment II), and the underlying mechanism of action (Experiment III). Fish exposed to CLEO concentrations reached a deep anesthesia stage in a concentration-dependent manner. However, the effects obtained during anesthetic induction were reversible, both in behavioral parameters and cardiac activity. The mechanism of action was observed after administration of flumazenil 1 mg/kg i.p., which showed greater resistance to loss of postural reflex and shorter latency for recovery. The mechanism of action indicates involvement of inhibitory GABA neurotransmission. The cardiac effects at the concentrations used were compatible with anesthesia, but no arrhythmias occurred that could compromise the hemodynamics of the fish, demonstrating its safety for short-duration anesthesia induction in \u003cem\u003eColossoma macropomum\u003c/em\u003e.\u003c/p\u003e","manuscriptTitle":"Behavioral, electrocardiographic evaluation, and mechanism of action of Curcuma longa essential oil in juvenile tambaqui Colossoma macropomum (Cuvier, 1818) subjected to immersion baths at different concentrations","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-14 14:02:12","doi":"10.21203/rs.3.rs-5154092/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"12074501-e435-49a5-a0e1-cdbe8a62266b","owner":[],"postedDate":"November 14th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-27T19:23:25+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-14 14:02:12","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5154092","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5154092","identity":"rs-5154092","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}