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Communication

Anaesthetic Effect of Clove Basil (Ocimum gratissimum) Essential Oil on Macrobrachium rosenbergii Post-Larvae

by
Cecília de Souza Valente
1,*,
Arielly Fávaro Mendes
2,
Caio Henrique do Nascimento Ferreira
3,
Bernardo Baldisserotto
4,
Berta Maria Heinzmann
5,
André Martins Vaz-dos-Santos
6 and
Eduardo Luis Cupertino Ballester
2,7
1
School of Biological & Chemical Sciences, University of Galway, H91TK33 Galway, Ireland
2
Graduate Program in Aquaculture Engineering, Federal University of Paraná, Palotina 85953-128, Brazil
3
Post Graduation Program in Aquaculture, Faculty of Agricultural and Veterinary Sciences, São Paulo State University, São Paulo 14884-900, Brazil
4
Department of Physiology and Pharmacology, Federal University of Santa Maria, Santa Maria 97105-900, Brazil
5
Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria 97105-900, Brazil
6
Sclerochronology Laboratory, Post Graduation Program in Aquaculture and Sustainable Development, Federal University of Paraná, Palotina 85953-128, Brazil
7
Shrimp Culture Laboratory, Post Graduation Program in Aquaculture and Sustainable Development, Federal University of Paraná, Palotina 85953-128, Brazil
*
Author to whom correspondence should be addressed.
Aquac. J. 2024, 4(3), 192-202; https://doi.org/10.3390/aquacj4030014
Submission received: 14 June 2024 / Revised: 12 August 2024 / Accepted: 9 September 2024 / Published: 12 September 2024
Browse Figure
Figure 1
<p>Polynomial curve (red line) and confidence intervals (blue lines) of the time (black dots) required to induce sedation (<b>A</b>), anaesthesia (<b>B</b>), and recovery (<b>C</b>) of <span class="html-italic">Macrobrachium rosenbergii</span> post-larvae exposed to different concentrations of <span class="html-italic">Ocimum gratissimum</span> essential oil. Confidence intervals for the standard deviation values of the polynomial standardized residuals in the sedation (<b>D</b>), anaesthesia (<b>E</b>), and recovery time (<b>F</b>); overlaps indicate statistical similarity (α = 0.05).</p> ">
Versions Notes

Abstract

:
This study evaluated the anaesthetic potential of clove basil (Ocimum gratissimum) essential oil (EO-OG) in the post-larvae (PLs) of M. rosenbergii. The PLs were individually transferred to aquariums (500 mL) containing 50, 100, 150, 200, or 300 μL L−1 EO-OG. A sixth group of PLs was exposed to freshwater only, and a seventh group was exposed to the highest concentration of ethanol used to dilute EO-OG. Upon reaching the anaesthesia stage, the PLs were transferred to aquariums (500 mL) with freshwater only to evaluate the recovery time. The shortest sedation times were observed at 200 (15.98 s) and 300 μL L−1 (25.85 s). The shortest anaesthesia time was observed at 200 μL L−1 (22.52 s). The longest recovery time was observed at 100 μL L−1 (1367.10 s); the shortest recovery time was observed at 150 μL L−1 (630.10 s). No mortality or adverse effects were observed in any experimental group. The concentration of 150 μL L−1 resulted in sedation (49.24 s), anaesthesia (80.10 s), and recovery (630.10 s) times within those suggested as adequate for freshwater prawn farming. This research demonstrated the sedative and anaesthetic properties of EO-OG and its potential application in prawn farming. This study recommends using 150 μL L−1 of EO-OG in M. rosenbergii PLs.

1. Introduction

The giant river prawn (Macrobrachium rosenbergii) stands out due to its high commercial value in crustacean farming. M. rosenbergii is the largest species in the genus Macrobrachium, family Palaemonidae. This species excels for its favourable characteristics in cultivation, such as easy reproduction in captivity, high fecundity, rusticity, and high resistance to diseases [1]. Freshwater prawns have the advantage of not depending on marine water, making it possible farm them in regions farther from the sea. Also, the production of M. rosenbergii presents a simple maturation and larviculture stage, with a production system compatible with small properties [2]. The world production of M. rosenbergii reached 294 thousand tons in 2020 [3], showing a considerable production increase compared to previous years and indicating an expanding market. Moreover, prawn farming has shown great growth in recent years, especially in emerging countries, thus playing an important socioeconomic role in income generation at regional and national levels [4].
In aquaculture systems, routine husbandry practices such as handling, biometric measurements, and transport can cause stress to farmed animals, negatively affecting their growth performance and welfare. To mitigate this impact, several torpor procedures (e.g., thermal shock) and anaesthetic substances (e.g., MS-222) are used in aquaculture. Nevertheless, hypothermia may cause inappropriate analgesia and anaesthesia [5]. Likewise, synthetic anaesthetics can cause adverse effects on both aquatic animals and farmers, as well as on the environment [6]. Given this, recent studies have sought alternatives to synthetic anaesthetics. Most of these alternative compounds also are sustainable and effective substitutes for conventional drugs and chemicals for disease control and prophylaxis in aquaculture [7]. In this context, the use of essential oils (EOs) as sedatives and natural anaesthetics has been shown to be efficient in mitigating the adverse effects during routine husbandry practices and as a sustainable alternative to synthetic compounds.
The EOs are natural volatile compounds formed by a complex mixture of low-molecular-weight volatile substances. They are secondary metabolites of aromatic plants and can be synthesized or extracted from different plant organs, including the leaves, flowers, stems, seeds, roots, and fruits [8]. The EOs are commonly limpid, but some can also be coloured; they are lipophilic and mainly formed by terpenes and terpenoids (major constituents), phenylpropanoids, and aromatic and aliphatic compounds [9]. EOs extracted from plants of the Ocimum genus have beneficial biological activities such as antimicrobial, antioxidant, anti-inflammatory, antinociceptive, analgesic, and immunomodulatory properties [10,11].
Ocimum gratissimum is an aromatic subshrub of the Lamiaceae family, popularly known as clove basil or scent leaf. It can reach one meter in height and is found in tropic and subtropical regions [12]. Its leaves are ovate-lanceolate, with double-toothed edges, membranous, and 4–8 cm long. It presents white inflorescences, arranged in erect paniculate racemes and, generally, in groups of three. It has small, capsule-type fruits with four spherical seeds [13]. The EO of Ocimum gratissimum (EO-OG) is mainly composed of eugenol, which is believed to be the major contributor to the biological activities of this EO [14,15]. The O. gratissimum plant is currently used as a medicinal plant to treat human illness and as a natural condiment and tea for human consumption [12,16,17]. In the aquaculture context, EO-OG has shown beneficial biological properties including an anaesthetic effect [18,19] and therapeutic uses [20,21]. Thereby, one anticipates the achievement of commercial use and formal authorization of EO-OG as an anaesthetic compound for aquaculture purposes.
In this context, this study aimed to answer the following question: Do natural anaesthetics such as essential oils have a sedative and anaesthetic effect on the PLs of freshwater prawns kept under optimum husbandry conditions? To answer the guiding question, the present work aimed to evaluate the potential sedative and anaesthetic effect of EO-OG on M. rosenbergii PLs, tested in five different concentrations.

2. Materials and Methods

2.1. Plant Origin and Essential Oil Extraction and Composition

The leaves of O. gratissimum were collected from the botanical collection of the Federal University of Santa Maria (UFSM, Brazil) Campus of Frederico Westphalen; the specimen was identified by Mr Adelino Filho and deposited in the Biology Department herbarium, UFSM, under voucher specimen number SMDB 11167. The EO was extracted according to de Souza Valente, dos Santos [18]. Briefly, the leafy material was extracted by hydro-distillation and, to separate the EO from the hydrolate, the mixture was placed in a separating funnel and was subjected to liquid–liquid partition with hexane P.A. A rotary evaporator, at 40 °C, was used to concentrate the hexane fraction and obtain the pure EO-OG. The extracted EO-OG was stored at 4 °C in an amber glass bottle until further analysis and use. The EO-OG composition was analysed by a gas chromatography-mass spectroscopy (GC-MS)–total ion chromatogram (TIC), and its compounds were identified using the Kováts retention index and a mass spectral library. The compounds identified were eugenol (88.46%), β-caryophyllene (5.30%), copaene (0.80%), E-β-ocimene (0.64%), and germacrene D (0.47%).

2.2. Experimental Design and Anaesthetic Trial

The post-larvae (PLs) of M. rosenbergii (0.0036 ± 0.005 g, total n = 280) were obtained from the Sustainable Aquaculture Research and Development Centre, Federal University of Paraná, Maripá, Paraná, Brazil. Water parameters were monitored twice a day and maintained at optimal levels for the farming of M. rosenbergii PLs throughout this study. The observed maximum and minimum values were as follows: temperature, 25.0–29.1 °C; pH, 7.75–8.24; and dissolved oxygen, 6.14–9.12 mg L−1. Survival was monitored twice a day throughout the anaesthetic trial.
The PLs were randomly collected from the larval hatchery and rearing tank and individually transferred to a 500 mL aquarium (temperature: 26.5–28.2 °C) containing one of the following concentrations of EO-OG (n = 40 PL concentration−1): 50, 100, 150, 200, and 300 μL L−1, previously diluted in ethanol P.A. (1:10). An additional experimental group of PLs was exposed to freshwater only (negative control), and another experimental group was exposed to the highest concentration of ethanol used to dilute the EO-OG (i.e., 2700 μL L−1; vehicle control). Each prawn was used only one time. Upon reaching the anaesthetic stage, the PLs were transferred to a 500 mL aquarium with freshwater only (without EO-OG) to evaluate the recovery time. The induction stage was observed for a maximum of 30 min. The sedation, anaesthesia, and recovery times were recorded in seconds (s) using a digital stopwatch.
The identification of the sedation, anaesthesia, and recovery stages was evaluated according to Coyle, Dasgupta [22]. The sedation stage was characterized by a partial loss of balance and reaction to touch stimuli. The anaesthesia stage was associated with a complete loss of balance and lack of reaction to touch stimuli. Finally, the recovery stage (in the aquarium with anaesthetic-free water) was considered complete when the prawn regained balance and reached the normal body posture, i.e., an upright position at the bottom of the aquarium.

2.3. Statistical Analyses

The time of each phase (sedation, anaesthesia, and recovery) was compared between the different concentrations of O. gratissimum essential oil using a polynomial contrast test [23,24]. A priori, linear (y = a + bx), quadratic (y = ax2 + bx + c), and cubic (y = ax3 + bx2 + cx + d) models were fitted, and the best fit was selected based on the lowest value of the Akaike information criterion [23,25]. The non-linear least squares method using the Levenberg–Marquardt algorithm with 9999 iterations was used to fit the models [26]. The standardized residuals were analysed; the data with |Z| > 1.96 were removed, and the models were re-fitted. The significance of the new standardized residuals was tested using Levene’s test [27]. The similarity between the different concentrations of O. gratissimum essential oil was assessed graphically by the intersection of the confidence intervals for the standard deviation values. A confidence level of 0.05 was used for all statistical procedures.

3. Results

Significant differences (p < 0.05) in the sedation, anaesthesia, and recovery times of M. rosenbergii PLs induced by varying concentrations of O. gratissimum essential oil were observed and properly described by cubic polynomial models (Figure 1, Table 1). Based on the results from the polynomial contrast test, the highest sedation time was observed at a concentration of 100 μL L−1 (41 s), being significantly different from all other values observed (Figure 1A), even though the concentration of 150 μL L−1 showed a higher arithmetic mean (49 s). The polynomial contrast test does not compare the means, and in a regression analysis, the reference mean is observed on the trend line. The arithmetic mean of the data at the concentration of 150 μL L−1 was higher due to presenting few low values, while the range of variation was large in the data at the concentration of 100 μL L−1.
The shortest sedation time was observed at a concentration of 200 μL L−1 (15 s) (Figure 1A). The concentrations of 100 and 200 μL L−1, with opposite behaviour, were significantly different from all the others (Figure 1D). The highest anaesthesia times were observed at 150 (80 s) and 100 μL L−1 (60 s), respectively (Figure 1B), which were similar (Figure 1E). Although the shortest induction time to the anaesthesia stage was observed at a concentration of 200 μL L−1 (22 s) (Figure 1B), it was statistically similar to the concentration of 50 μL L−1 (54 s) (Figure 1E). The longest recovery time was observed at a concentration of 100 μL L−1 (1367 s), followed by the concentration of 50 μL L−1 (1006 s), from which it did not differ (Figure 1C,F). The shortest recovery time was observed at a concentration of 150 μL L−1 (630 s) (Figure 1C), significantly different from the lower concentrations but with an unclear trend in relation to higher concentrations (Figure 1F). In all phases analysed, the models showed a trough close to the 250 μL L−1 concentration, an effect of the polynomial modelling, as this concentration was not tested.
All induction and recovery times were within those deemed adequate for prawn anaesthesia (Table 2). The concentration of 150 μL L−1, followed by the concentration of 200 μL L−1, resulted in sedation (49 s and 15 s), anaesthesia (80 s and 22 s), and recovery times (630 s and 829 s) appropriate for the PL management of M. rosenbergii. The EO-OG concentrations evaluated did not cause any death or side effects during the anaesthesia trial. Likewise, no mortality was observed in the control groups. The ethanol P.A. did not cause any sedative or anaesthetic effect on M. rosenbergii PLs at the concentration tested.

4. Discussion

Increasing attention has been paid to the welfare of decapod crustaceans worldwide. In 2022, the United Kingdom included decapod crustaceans in its Animal Welfare (Sentience) Bill, thus legally acknowledging the sentience of these animals, alongside vertebrates and cephalopods [28]. One anticipates that countries in the European Union, the USA, and Brazil, will also advance their protective animal legislation and include decapods in their legislative scope. Thereby, it is expected that an increased protection of animal interests will lead to improvements in the welfare of farmed decapods. Accommodating these advances in practical and applied knowledge will improve aquaculture. For example, attention is increasing on how decapod crustaceans are maintained in the sphere of freshwater prawn farming. Management practices (e.g., biometrics and transport) can be refined to improve growth performance, increase survival, and facilitate animal handling. Also, the refinement of practices with animals positively influences the results of scientific studies.
Decapod crustaceans respond to noxious stimuli and experience pain and stress [29,30,31]. Therefore, the use of analgesics and anaesthetics is recommended to ensure animal welfare, suppress nociception, reduce animal stress, and facilitate husbandry practices. Natural anaesthetics such as EO-OG could aid in practices that aim for low-stress methods when handling, transporting, and sampling prawns. A recent study conducted by our research group has validated the use of EO-OG as a potential plant-derived anaesthetic for M. rosenbergii juveniles; it recommended the concentration of 400 μL L−1, with the use of mild alkaline water [18]. Similarly, the EO-OG induces adequate anaesthesia in pink shrimp (Farfantepenaeus paulensis; 100 μL L−1) [32]; pacamã catfish (Lophiosilurus alexandri; 90 and 150 mg L−1) [33]; matrinxã fish (Brycon amazonicus; 20 to 60 mg L−1) [34]; Brazilian flounder (Paralichthys orbignyanus; 10 mg L−1 for sedation, 50 to 100 mg L−1 for anaesthesia) [35]; Nile tilapia (Oreochromis niloticus; 5 mg L−1 for sedation, 90 to 150 mg L−1 for anaesthesia) [36], cachara catfish (Pseudoplatystoma reticulatum; 187 mL L−1) [37], and silver catfish (Rhamdia quelen; 150 to 300 mg L−1) [19]. The additional biological properties of EO-OG in aquatic animals include anthelmintic activity [20], antiparasitic properties [21], antioxidant effect [21,38], and immunostimulant capabilities [38].
The biological properties of EO-OG can be attributed to its bioactive compounds, particularly its major constituent, i.e., eugenol, in a potentiation, additive, or synergic effect with minor constituents, such as β-caryophyllene, copaene, E-β-ocimene, and germacrene D [16,18,39]. Eugenol has been largely used as an effective plant-derived anaesthetic for decapod crustaceans, particularly through immersion, including as a sedative for transportation, to induce surgical anaesthesia, and for euthanasia by overdose [40]. The main mechanism of action of eugenol as an anaesthetic is attributed to the inhibition of nerve impulses by blocking selective channels, particularly voltage-gated sodium, potassium, and calcium channels, with an effect on nociceptive and non-nociceptive nerve fibres [5,41,42]. Another mechanism that contributes to eugenol’s sedative and anaesthetic effect is the inhibition of synaptic transmission through enhancing the activity of the GABAA receptors [41,42]. Based on results from clove oil, whose main compounds are eugenol and isoeugenol, the main biodistribution and excretion vias of eugenol may include the intestinal tract and the gills [43].
Similarly, the minor compounds of EO-OG may also contribute to the EO-OG anaesthetic effect. For instance, β-caryophyllene is a sesquiterpene with known antinociceptive and anaesthetic effects in humans [44], rats [45], mice [46], fish [47], and crustaceans [18]; it is present in EOs such as lemon verbena (Aloysia triphylla) [48], candeia (Eremanthus erythropappus) [49], catnip (Nepeta cataria) [50], thyme (Thymus vulgaris) [47], rosemary (Rosmarinus officinalis), oregano (Origanum vulgare), basil (Ocimum sp.), ginger (Zingiber officinale), and clove (Syzygium aromaticum) [51]. Likewise, copaene is a sesquiterpene present in different EOs with sedative and anaesthetic effects in aquatic animals, such as EOs of basil (Ocimum sp.) and bushy lippia (Lippia alba) [52,53,54], and geranium oil (Pelargonium graveolens) [55]. Also, ocimene, a monoterpene, is one of the compounds of EOs with potential for aquatic animals, such as basil (Ocimum sp.) [52,53,56], lavender (Lavandula angustifolia) [57], and piper (Piper divaricatum) [58]. Equally, germacrene is a sesquiterpene and can be found in EOs such as geranium oil (P. graveolens) [55], piper (P. divaricatum) [58], black cinnamon (Nectandra megapotamica) [59], and ginger (Zingiber officinale) [37].
In the present study, the sedation and anaesthesia induction times were directly influenced by the concentration of EO-OG used. That is, the anaesthetic effect of EO-OG was concentration dependent, i.e., increasing concentrations of EO-OG led to reduced induction times, particularly above 200 μL L−1. Previous studies showed that many plant-derived anaesthetics, including EO-OG, lead to sedation and anaesthesia in aquatic animals in a concentration-dependent manner [18,19,32,35]. A parallel with the concentration effect observed in inhalation anaesthesia in humans might be hypothesized. The concentration effect is determined by the ratio between the alveolar concentration (FA) and the inspired concentration (FI), observed during the initial administration of inhaled anaesthetics with nitrous oxide [60]. The higher the inspired concentration of the anaesthetic gas, the faster the FA/FI ratio increases, which leads to an anaesthetic induction acceleration [61,62]. Likewise, in humans, volatile anaesthetics cause a concentration-dependent response in terms of anaesthetic effects. That is, the amnesia, sedation, unconsciousness, and immobilization effects are concentration dependent, in the order of sensitiveness of the anaesthetic endpoints [63]. Similarly, a threshold for the anaesthetic activity to EO-OG may be theorized. The shortest sedation and anaesthesia induction times were observed at the second highest tested concentration (i.e., 200 µL L−1), suggesting the anaesthetic activity of EO-OG is concentration dependent, although after a threshold, its bioactivity may decrease. Future studies may investigate whether an analogous concentration effect and a concentration-dependent response, with a threshold, of the anaesthetic triad also occur in decapod crustaceans. This might relate to the gills, the gas exchange ratio by decapods’ cardiovascular system, and the respiratory pigment haemocyanin.
Likewise, the life stage of M. rosenbergii may influence the anaesthetic effectiveness of EO-OG. When comparing the results of the present study with a recent work of our group [18], a lower concentration (i.e., half to less than half concentration) and a lower induction time (i.e., half time) were necessary to induce sedation and anaesthesia in M. rosenbergii PLs in comparison to M. rosenbergii juveniles. Similar results were observed in the Chinese grass shrimp (Palaemonetes sinensis) anaesthetised with eugenol. Based on their body size, smaller P. sinensis presented a shorter induction time than larger P. sinensis [64]. Likewise, the PLs of the Pacific white shrimp (Penaeus vannamei) required a lower anaesthetic concentration than sub-adult animals, when anaesthetized with eugenol (175 and 200 µL L−1, respectively) and Lippia alba EO (500 and 750 µL L−1, respectively) [65]. Differences in the gill surface area may be one way to understand this variation in anaesthesia response. The smaller the body size, the larger the gill surface area; thus, smaller animals (e.g., PLs compared to juveniles) have higher anaesthetic absorption rates than larger animals [32]. Likewise, similar to what is observed in fish, the metabolism may also influence the response variation. The high basal metabolic rate of small animals results in a high consumption of oxygen [6], leading to a greater absorption of the anaesthetic compound present in the water bath.
To be convenient for the aquaculture context, an adequate anaesthetic ideally would induce anaesthesia within three to five minutes, allow a smooth recovery time of up to ten minutes, and not cause any adverse effect [66,67]. In the present study, the EO-OG concentration of 150 μL L−1, followed by the concentration of 200 μL L−1, resulted in adequate sedation, anaesthesia, and recovery times for M. rosenbergii PLs, without any adverse effect. Considering a potential predilection for using the lowest possible effective concentration of EO-OG, both in terms of cost and management, and a short recovery time, the concentration of 150 μL L−1 of EO-OG proved to be the most recommended among the concentrations studied. Management practices in prawn farming, such as capture, handling, sampling, and transport, can cause animal stress and loss of production, impacting prawn welfare and leading to eventual mortality. These practices can be facilitated using natural anaesthetics such as EO-OG, potentially facilitating animal handling and preserving the prawns’ welfare.

5. Conclusions

Based on the results obtained in this study, the recommended concentration of O. gratissimum essential oil to be used in routine freshwater prawn husbandry, such as handling, sampling, and transport of M. rosenbergii PLs, is 150 μL L−1. This concentration leads to sedation and anaesthesia induction within 49.24 and 80.10 s, respectively, and recovery within 630.10 s, without causing any adverse effect, thus being suggested as suitable for use in prawn farming. Further research will benefit from studying the bioactive compounds of EO-OG and their mechanism of action as a plant-derived anaesthetic for prawns.

Author Contributions

Conceptualization, E.L.C.B.; methodology, E.L.C.B.; formal analysis, C.d.S.V., A.F.M., C.H.d.N.F., B.B., B.M.H. and A.M.V.-d.-S.; investigation, A.F.M. and C.H.d.N.F.; resources, B.B. and E.L.C.B.; data curation, A.F.M. and C.H.d.N.F.; writing—original draft preparation, C.d.S.V. and A.F.M.; writing—review and editing, C.d.S.V., A.F.M., C.H.d.N.F., B.B., B.M.H., A.M.V.-d.-S. and E.L.C.B.; visualization, C.d.S.V. and A.M.V.-d.-S.; supervision, E.L.C.B.; project administration, E.L.C.B.; funding acquisition, E.L.C.B. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the National Council of Technological and Scientific Development (CNPq), Brazil, under grant No. PQ 311456/2020-0, to Eduardo Luis Cupertino Ballester. Eduardo Luis Cupertino Ballester (No. PQ 311456/2020-0), André Martins Vaz-dos-Santos (No. CNPq 308082/2022-2), and Bernardo Baldisserotto (No. PQ 301816/2022-2) are recipients of CNPq productivity research grants.

Institutional Review Board Statement

Ethical review and approval were waived for this study because the current Brazilian animal protective legislation (Law No. 11,794, Brazil) does not require ethical approval by an Animal Welfare Committee when the study encompasses the use of decapod crustaceans. This study was conducted in a research facility from a Brazilian federal university. This study followed the recommended commercial and academic parameters for prawn farming, including adequate feeding and water quality. Prawns were euthanized following torpor with thermal shock.

Data Availability Statement

All data that support the findings of this study are available in this manuscript.

Conflicts of Interest

This study was part of Ms. Arielly Fávaro Mendes’s graduation thesis. The authors declare no additional conflicts of interest.

References

  1. Cavallo, R.O.; Lavens, P.; Sorgeloos, P. Reproductive Performance of Macrobrachium rosenbergii Females in Captivity. J. World Aquac. Soc. 2001, 32, 60–67. [Google Scholar] [CrossRef]
  2. Barros, H.P.; Valenti, W.C. Comportamento alimentar do camarão de água doce, Macrobrachium rosenbergii (De Man) (Crustacea, Palaemonidae) durante a fase larval: Análise qualitativa. Rev. Bras. Zool. 1997, 14, 785–793. [Google Scholar] [CrossRef]
  3. FAO. The State of World Fisheries and Aquaculture 2022: Towards Blue Transformation. In The State of World Fisheries and Aquaculture (SOFIA); Food and Agricultural Organization of the United Nations: Rome, Italy, 2022; p. 266. [Google Scholar]
  4. Machado, I.D.S.; Nunes, C.A.R.; dos Santos, H.B.L.; Lima, J.A.; Meira, T.M.; Da silva Sena, E.; da Silva, W.N. Desempenho do camarão Macrobrachium amazonicum (Heller, 1862) (Crustacea: Decapoda: Palaemonidae), em diferentes densidades. Rev. Bras. Eng. Pesca 2018, 11, 29–37. [Google Scholar] [CrossRef]
  5. de Souza Valente, C. Anaesthesia of decapod crustaceans. Vet. Anim. Sci. 2022, 16, 100252. [Google Scholar] [CrossRef] [PubMed]
  6. Zahl, I.H.; Samuelsen, O.; Kiessling, A. Anaesthesia of farmed fish: Implications for welfare. Fish Physiol. Biochem. 2012, 38, 201–218. [Google Scholar] [CrossRef] [PubMed]
  7. Aydın, B.; Barbas, L.A.L. Sedative and anesthetic properties of essential oils and their active compounds in fish: A review. Aquaculture 2020, 520, 734999. [Google Scholar] [CrossRef]
  8. Khan, S.; Sahar, A.; Tariq, T.; Sameen, A.; Tariq, F. Chapter 1—Essential oils in plants: Plant physiology, the chemical composition of the oil, and natural variation of the oils (chemotaxonomy and environmental effects, etc.). In Essential Oils; Nayik, G.A., Ansari, M.J., Eds.; Academic Press: Cambridge, MA, USA, 2023; pp. 1–36. [Google Scholar] [CrossRef]
  9. Bakkali, F.; Averbeck, S.; Averbeck, D.; Idaomar, M. Biological effects of essential oils—A review. Food Chem. Toxicol. 2008, 46, 446–475. [Google Scholar] [CrossRef]
  10. Pandey, A.K.; Singh, P.; Tripathi, N.N. Chemistry and bioactivities of essential oils of some Ocimum species: An overview. Asian Pac. J. Trop. Biomed. 2014, 4, 682–694. [Google Scholar] [CrossRef]
  11. Avetisyan, A.; Markosian, A.; Petrosyan, M.; Sahakyan, N.; Babayan, A.; Aloyan, S.; Trchounian, A. Chemical composition and some biological activities of the essential oils from basil Ocimum different cultivars. BMC Complement. Altern. Med. 2017, 17, 60. [Google Scholar] [CrossRef]
  12. Mbegbu, N.; Nwajinka, C.; Amaefule, D. Thin layer drying models and characteristics of scent leaves (Ocimum gratissimum) and lemon basil leaves (Ocimum africanum). Heliyon 2021, 7, e05945. [Google Scholar] [CrossRef]
  13. Lorenzi, H.; Matos, F.J.d.A.; Cavalleiro, A.d.S.; Brochini, V.F.; Souza, V.C. Plantas Medicinais no Brasil: Nativas e Exóticas, 3rd ed.; Instituo Plantarum: Nova Odessa, Brazil, 2021; 576p. [Google Scholar]
  14. Mahapatra, S.K.; Roy, S. Phytopharmacological approach of free radical scavenging and anti-oxidative potential of eugenol and Ocimum gratissimum Linn. Asian Pac. J. Trop. Med. 2014, 7, S391–S397. [Google Scholar] [CrossRef] [PubMed]
  15. Priyanka, C.; Shivika, S.; Vikas, S. Ocimum gratissimum: A Review on Ethnomedicinal Properties, Phytochemical Constituents, and Pharmacological Profile. In Biotechnological Approaches for Medicinal and Aromatic Plants: Conservation, Genetic Improvement and Utilization; Kumar, N., Ed.; Springer: Singapore, 2018; pp. 251–270. [Google Scholar] [CrossRef]
  16. Ugbogu, O.C.; Emmanuel, O.; Agi, G.O.; Ibe, C.; Ekweogu, C.N.; Ude, V.C.; Uche, M.E.; Nnanna, R.O.; Ugbogu, E.A. A review on the traditional uses, phytochemistry, and pharmacological activities of clove basil (Ocimum gratissimum L.). Heliyon 2021, 7, e08404. [Google Scholar] [CrossRef] [PubMed]
  17. Di Stasi, L.C.; Oliveira, G.P.; Carvalhaes, M.A.; Queiroz, M., Jr.; Tien, O.S.; Kakinami, S.H.; Reis, M.S. Medicinal plants popularly used in the Brazilian Tropical Atlantic Forest. Fitoterapia 2002, 73, 69–91. [Google Scholar] [CrossRef] [PubMed]
  18. de Souza Valente, C.; dos Santos, G.; Becker, A.G.; Heinzmann, B.M.; Caron, B.O.; Baldisserotto, B.; Ballester, E.L.C. Anaesthetic effect of clove basil (Ocimum gratissimum L.) essential oil on the giant river prawn (Macrobrachium rosenbergii, De Man 1879) exposed to different water pHs. Aquac. Int. 2024, 32, 1493–1505. [Google Scholar] [CrossRef]
  19. Silva, L.d.L.; Parodi, T.V.; Reckziegel, P.; Garcia, V.d.O.; Bürger, M.E.; Baldisserotto, B.; Malmann, C.A.; Pereira, A.M.S.; Heinzmann, B.M. Essential oil of Ocimum gratissimum L.: Anesthetic effects, mechanism of action and tolerance in silver catfish, Rhamdia quelen. Aquaculture 2012, 350–353, 91–97. [Google Scholar] [CrossRef]
  20. Meneses, J.; do Couto, M.; Sousa, N.; Cunha, F.d.S.; Abe, H.; Ramos, F.; Chagas, E.; Chaves, F.; Martins, M.; Maria, A. Efficacy of Ocimum gratissimum essential oil against the monogenean Cichlidogyrus tilapiae gill parasite of Nile tilapia. Arq. Bras. Med. Veterinária Zootec. 2018, 70, 497–504. [Google Scholar] [CrossRef]
  21. Bandeira Jr, G.; Pês, T.S.; Saccol, E.M.; Sutili, F.J.; Rossi Jr, W.; Murari, A.L.; Heinzmann, B.M.; Pavanato, M.A.; de Vargas, A.C.; Silva, L.d.L. Potential uses of Ocimum gratissimum and Hesperozygis ringens essential oils in aquaculture. Ind. Crops Prod. 2017, 97, 484–491. [Google Scholar] [CrossRef]
  22. Coyle, S.D.; Dasgupta, S.; Tidwell, J.H.; Beavers, T.; Bright, L.A.; Yasharian, D.K. Comparative efficacy of anesthetics for the freshwater prawn Macrobrachiurn rosenbergii. J. World Aquac. Soc. 2005, 36, 282–290. [Google Scholar] [CrossRef]
  23. Yossa, R.; Verdegem, M. Misuse of multiple comparison tests and underuse of contrast procedures in aquaculture publications. Aquaculture 2015, 437, 344–350. [Google Scholar] [CrossRef]
  24. de Souza Valente, C.; Coates, C.J.; Cagol, L.; Bombardelli, R.A.; Becker, A.G.; Schmidt, D.; Heinzmann, B.M.; Vaz-dos-Santos, A.M.; Baldisserotto, B.; Ballester, E.L.C. Antioxidant status and performance of Macrobrachium rosenbergii juveniles fed diets containing non-nutritive Aloysia triphylla essential oil. Aquac. Int. 2024, 1–14. [Google Scholar] [CrossRef]
  25. Zuur, A.F.; Ieno, E.N.; Walker, N.J.; Saveliev, A.A.; Smith, G.M. Mixed Effects Models and Extensions in Ecology with R; Springer: Berlin/Heidelberg, Germany, 2009; Volume 574. [Google Scholar]
  26. Motulsky, H.; Christopoulos, A. Fitting Models to Biological Data Using Linear and Nonlinear Regression: A Practical Guide to Curve Fitting; Oxford University Press: Oxford, UK, 2004. [Google Scholar]
  27. NIST/SEMATECH. Engineering Statistics: E-Handbook of Statistical Methods; NIST/SEMATECH: Gaithersburg, MD, USA, 2024. [CrossRef]
  28. Parliament, U. Animal Welfare (Sentience) Bill [HL]. In Bill 255; Parliament, U., Ed.; House of Lords: London, UK, 2022. [Google Scholar]
  29. Gherardi, F. Behavioural indicators of pain in crustacean decapods. Ann. Dell’istituto Super. Sanità 2009, 45, 432–438. [Google Scholar] [CrossRef] [PubMed]
  30. Elwood, R.W. Potential pain in fish and decapods: Similar experimental approaches and similar results. Front. Vet. Sci. 2021, 8, 631151. [Google Scholar] [CrossRef] [PubMed]
  31. Crump, A.; Browning, H.; Schnell, A.; Burn, C.; Birch, J. Sentience in decapod crustaceans: A general framework and review of the evidence. Anim. Sentience 2022, 7, 1. [Google Scholar] [CrossRef]
  32. Becker, A.J.; Vaz, L.J.; Garcia, L.d.O.; Wasielesky Jr, W.; Heinzmann, B.M.; Baldisserotto, B. Anesthetic potential of different essential oils for two shrimp species, Farfantepenaeus paulensis and Litopenaeus vannamei (Decapoda, Crustacea). Ciência Rural 2021, 51, e20200793. [Google Scholar] [CrossRef]
  33. Boaventura, T.P.; Souza, C.F.; Ferreira, A.L.; Favero, G.C.; Baldissera, M.D.; Heinzmann, B.M.; Baldisserotto, B.; Luz, R.K. Essential oil of Ocimum gratissimum (Linnaeus, 1753) as anesthetic for Lophiosilurus alexandri: Induction, recovery, hematology, biochemistry and oxidative stress. Aquaculture 2020, 529, 735676. [Google Scholar] [CrossRef]
  34. Ribeiro, A.S.; Batista, E.d.S.; Dairiki, J.K.; Chaves, F.C.M.; Inoue, L.A.K.A. Anesthetic properties of Ocimum gratissimum essential oil for juvenile matrinxã. Acta Scientiarum. Anim. Sci. 2016, 38, 1–7. [Google Scholar] [CrossRef]
  35. Benovit, S.C.; Gressler, L.T.; de Lima Silva, L.; de Oliveira Garcia, L.; Okamoto, M.H.; dos Santos Pedron, J.; Sampaio, L.A.; Rodrigues, R.V.; Heinzmann, B.M.; Baldisserotto, B. Anesthesia and transport of Brazilian flounder, Paralichthys orbignyanus, with essential oils of Aloysia gratissima and Ocimum gratissimum. J. World Aquac. Soc. 2012, 43, 896. [Google Scholar] [CrossRef]
  36. Ferreira, A.L.; Favero, G.C.; Boaventura, T.P.; de Freitas Souza, C.; Ferreira, N.S.; Descovi, S.N.; Baldisserotto, B.; Heinzmann, B.M.; Luz, R.K. Essential oil of Ocimum gratissimum (Linnaeus, 1753): Efficacy for anesthesia and transport of Oreochromis niloticus. Fish Physiol. Biochem. 2021, 47, 135–152. [Google Scholar] [CrossRef]
  37. da Silva, L.A.; Martins, M.A.; Santo, F.E.; Oliveira, F.C.; Chaves, F.C.M.; Chagas, E.C.; Martins, M.L.; de Campos, C.M. Essential oils of Ocimum gratissimum and Zingiber officinale as anesthetics for the South American catfish Pseudoplatystoma reticulatum. Aquaculture 2020, 528, 735595. [Google Scholar] [CrossRef]
  38. Abdel-Tawwab, M.; Adeshina, I.; Jenyo-Oni, A.; Ajani, E.K.; Emikpe, B.O. Growth, physiological, antioxidants, and immune response of African catfish, Clarias gariepinus (B.), to dietary clove basil, Ocimum gratissimum, leaf extract and its susceptibility to Listeria monocytogenes infection. Fish Shellfish Immunol. 2018, 78, 346–354. [Google Scholar] [CrossRef]
  39. Efferth, T.; Koch, E. Complex Interactions between Phytochemicals. The Multi-Target Therapeutic Concept of Phytotherapy. Curr. Drug Targets 2011, 12, 122–132. [Google Scholar] [CrossRef] [PubMed]
  40. Spoors, F.; James, M.A.; Mendo, T.; McKnight, J.C.; Bønnelycke, E.-M.S.; Khan, N. Investigating clove oil and its derivatives as anaesthetic agents for decapod crustaceans to improve welfare commercially and at slaughter. Front. Anim. Sci. 2023, 4, 1180977. [Google Scholar] [CrossRef]
  41. Lehotzky, D.; Eske, A.I.; Zupanc, G.K.H. The effect of eugenol anesthesia on the electric organ discharge of the weakly electric fish Apteronotus leptorhynchus. Fish Physiol. Biochem. 2023, 49, 1321–1338. [Google Scholar] [CrossRef] [PubMed]
  42. Chung, G.; Oh, S.B. Eugenol as Local Anesthetic. In Natural Products: Phytochemistry, Botany and Metabolism of Alkaloids, Phenolics and Terpenes; Ramawat, K.G., Mérillon, J.-M., Eds.; Springer: Berlin/Heidelberg, Germany, 2013; pp. 4001–4015. [Google Scholar] [CrossRef]
  43. Kaewmalun, S.; Yata, T.; Kitiyodom, S.; Yostawonkul, J.; Namdee, K.; Kamble, M.T.; Pirarat, N. Clove Oil-Nanostructured Lipid Carriers: A Platform of Herbal Anesthetics in Whiteleg Shrimp (Penaeus vannamei). Foods 2022, 11, 3162. [Google Scholar] [CrossRef]
  44. Fidyt, K.; Fiedorowicz, A.; Strządała, L.; Szumny, A. β-caryophyllene and β-caryophyllene oxide—Natural compounds of anticancer and analgesic properties. Cancer Med. 2016, 5, 3007–3017. [Google Scholar] [CrossRef]
  45. Ghelardini, C.; Galeotti, N.; Di Cesare Mannelli, L.; Mazzanti, G.; Bartolini, A. Local anaesthetic activity of β-caryophyllene. Il Farm. 2001, 56, 387–389. [Google Scholar] [CrossRef]
  46. Moghrovyan, A.; Parseghyan, L.; Sevoyan, G.; Darbinyan, A.; Sahakyan, N.; Gaboyan, M.; Karabekian, Z.; Voskanyan, A. Antinociceptive, anti-inflammatory, and cytotoxic properties of Origanum vulgare essential oil, rich with β-caryophyllene and β-caryophyllene oxide. Korean J Pain 2022, 35, 140–151. [Google Scholar] [CrossRef]
  47. Boaventura, T.P.; dos Santos, F.A.C.; de Sena Souza, A.; Batista, F.S.; Júlio, G.S.C.; Luz, R.K. Thymol and linalool chemotypes of the essential oil of Thymus vulgaris (thyme) as anesthetic for Colossoma macropomum: Physiology and feed consumption. Aquaculture 2022, 554, 738161. [Google Scholar] [CrossRef]
  48. Kim, N.S.; Lee, D.S. Headspace solid-phase microextraction for characterization of fragrances of lemon verbena (Aloysia triphylla) by gas chromatography-mass spectrometry. J. Sep. Sci. 2004, 27, 96–100. [Google Scholar] [CrossRef]
  49. Sousa, O.V.; Silvério, M.S.; Del-Vechio-Vieira, G.; Matheus, F.C.; Yamamoto, C.H.; Alves, M.S. Antinociceptive and anti-inflammatory effects of the essential oil from Eremanthus erythropappus leaves. J. Pharm. Pharmacol. 2008, 60, 771–777. [Google Scholar] [CrossRef]
  50. de Souza, A.d.S.L.; Peret, A.C.; Hamoy, M.; de Souza, R.A.L.; Torres, M.F.; Barbas, L.A.L. Propofol and essential oil of Nepeta cataria induce anaesthesia and marked myorelaxation in tambaqui Colossoma macropomum: Implications on cardiorespiratory responses. Aquaculture 2019, 500, 160–169. [Google Scholar] [CrossRef]
  51. Francomano, F.; Caruso, A.; Barbarossa, A.; Fazio, A.; La Torre, C.; Ceramella, J.; Mallamaci, R.; Saturnino, C.; Iacopetta, D.; Sinicropi, M.S. β-Caryophyllene: A Sesquiterpene with Countless Biological Properties. Appl. Sci. 2019, 9, 5420. [Google Scholar] [CrossRef]
  52. da Silva, E.; Aldegunde, M.; da Silva, D.F.; Lopes, C.; Bertoldi, F.C.; Weber, R.A. Assessment of induction and recovery times of anaesthesia in Astyanax bimaculatus using 2-phenoxyethanol and the essential oils of Melaleuca alternifolia and Ocimum gratissimum. Aquac. Res. 2020, 51, 577–583. [Google Scholar] [CrossRef]
  53. Rucinque, D.S.; Ferreira, P.F.; Leme, P.R.P.; Lapa-Guimarães, J.; Viegas, E.M.M. Ocimum americanum and Lippia alba essential oils as anaesthetics for Nile tilapia: Induction, recovery of apparent unconsciousness and sensory analysis of fillets. Aquaculture 2021, 531, 735902. [Google Scholar] [CrossRef]
  54. Yigit, N.O.; Metin, S.; Sabuncu, O.F.; Didinen, B.I.; Didinen, H.; Ozmen, O.; Koskan, O. Efficiency of Ocimum basilicum and Eucalyptus globulus essential oils on anesthesia and histopathology of rainbow trout, Oncorhynchus mykiss. J. World Aquac. Soc. 2022, 53, 1051–1061. [Google Scholar] [CrossRef]
  55. Can, E.; Kizak, V.; Seyhaneyildiz Can, Ş.; Özçiçek, E. Anesthetic potential of geranium (Pelargonium graveolens) oil for two cichlid species, Sciaenochromis fryeri and Labidochromis caeruleus. Aquaculture 2018, 491, 59–64. [Google Scholar] [CrossRef]
  56. de Lima Boijink, C.; Queiroz, C.A.; Chagas, E.C.; Chaves, F.C.M.; Inoue, L.A.K.A. Anesthetic and anthelminthic effects of clove basil (Ocimum gratissimum) essential oil for tambaqui (Colossoma macropomum). Aquaculture 2016, 457, 24–28. [Google Scholar] [CrossRef]
  57. Yigit, N.O.; Metin, S.; Didinen, B.I.; Didinen, H.; Ozmen, O. Effect of lavander (Lavandula angustifolia) and laurel (Laurus nobilis) essential oils as anesthesics in rainbow trout (Oncorhynchus mykiss). Aquaculture 2022, 557, 738328. [Google Scholar] [CrossRef]
  58. Vilhena, C.S.; do Nascimento, L.A.S.; de Aguiar Andrade, E.H.; da Silva, J.K.d.R.; Hamoy, M.; Torres, M.F.; Barbas, L.A.L. Essential oil of Piper divaricatum induces a general anaesthesia-like state and loss of skeletal muscle tonus in juvenile tambaqui, Colossoma macropomum. Aquaculture 2019, 510, 169–175. [Google Scholar] [CrossRef]
  59. Ferraz, E.d.O.; Vieira, M.A.R.; Ferreira, M.I.; Fernandes Junior, A.; Marques, M.O.M.; Minatel, I.O.; Albano, M.; Sambo, P.; Lima, G.P.P. Seasonality effects on chemical composition, antibacterial activity and essential oil yield of three species of Nectandra. PLoS ONE 2018, 13, e0204132. [Google Scholar] [CrossRef]
  60. Myatt, J. Pharmacology of Inhalational Anaesthetics. Update Anaesth. Educ. Anaesth. Worldw. 2008, 24, 102–107. [Google Scholar]
  61. Khan, K.S.; Hayes, I.; Buggy, D.J. Pharmacology of anaesthetic agents II: Inhalation anaesthetic agents. Contin. Educ. Anaesth. Crit. Care Pain 2013, 14, 106–111. [Google Scholar] [CrossRef]
  62. Wouden, J.; Miller, K.W. General anesthetic pharmacology. In Principles of Pharmacology: The Pathophysiologic Basis of Drug Therapy, 3rd ed.; Lippincott Williams & Wilkins: Philadelphia, PA, USA, 2012; pp. 240–259. [Google Scholar]
  63. Hudson, A.E.; Herold, K.F.; Hemmings, H.C. Chapter 10—Pharmacology of Inhaled Anesthetics. In Pharmacology and Physiology for Anesthesia; Hemmings, H.C., Egan, T.D., Eds.; W.B. Saunders: Philadelphia, PA, USA, 2013; pp. 159–179. [Google Scholar] [CrossRef]
  64. Li, Y.; She, Q.; Han, Z.; Sun, N.; Liu, X.; Li, X. Anaesthetic Effects of Eugenol on Grass Shrimp (Palaemonetes sinensis) of Different Sizes at Different Concentrations and Temperatures. Sci. Rep. 2018, 8, 11007. [Google Scholar] [CrossRef] [PubMed]
  65. Parodi, T.V.; Cunha, M.A.; Heldwein, C.G.; de Souza, D.M.; Martins, Á.C.; Garcia, L.d.O.; Junior, W.W.; Monserrat, J.M.; Schmidt, D.; Caron, B.O. The anesthetic efficacy of eugenol and the essential oils of Lippia alba and Aloysia triphylla in post-larvae and sub-adults of Litopenaeus vannamei (Crustacea, Penaeidae). Comp. Biochem. Physiol. Part C Toxicol. Pharmacol. 2012, 155, 462–468. [Google Scholar] [CrossRef] [PubMed]
  66. Bodur, T.; Afonso, J.M.; Montero, D.; Navarro, A. Assessment of effective dose of new herbal anesthetics in two marine aquaculture species: Dicentrarchus labrax and Argyrosomus regius. Aquaculture 2018, 482, 78–82. [Google Scholar] [CrossRef]
  67. Marking, L.L.; Meyer, F.P. Are Better Anesthetics Needed in Fisheries? Fisheries 1985, 10, 2–5. [Google Scholar] [CrossRef]
Aquacj 04 00014 g001
Figure 1. Polynomial curve (red line) and confidence intervals (blue lines) of the time (black dots) required to induce sedation (A), anaesthesia (B), and recovery (C) of Macrobrachium rosenbergii post-larvae exposed to different concentrations of Ocimum gratissimum essential oil. Confidence intervals for the standard deviation values of the polynomial standardized residuals in the sedation (D), anaesthesia (E), and recovery time (F); overlaps indicate statistical similarity (α = 0.05).
Figure 1. Polynomial curve (red line) and confidence intervals (blue lines) of the time (black dots) required to induce sedation (A), anaesthesia (B), and recovery (C) of Macrobrachium rosenbergii post-larvae exposed to different concentrations of Ocimum gratissimum essential oil. Confidence intervals for the standard deviation values of the polynomial standardized residuals in the sedation (D), anaesthesia (E), and recovery time (F); overlaps indicate statistical similarity (α = 0.05).
Aquacj 04 00014 g001
Table 1. Coefficient (a, b, c, d) and standard error (S.E.) values of the cubic polynomials fitted for the time required to induce sedation, anaesthesia, and recovery in Macrobrachium rosenbergii post-larvae exposed to different concentrations of Ocimum gratissimum essential oil (F: model statistics, p: probability value, and standard error of the model).
Table 1. Coefficient (a, b, c, d) and standard error (S.E.) values of the cubic polynomials fitted for the time required to induce sedation, anaesthesia, and recovery in Macrobrachium rosenbergii post-larvae exposed to different concentrations of Ocimum gratissimum essential oil (F: model statistics, p: probability value, and standard error of the model).
Modela±S.E.b±S.E.c±S.E.d±S.E.Fp-ValueS.E. Regression
Sedation time1.862 × 10−50.0000−0.0090.00111.2480.16571.79566.808673.0462.59 × 10−3110.13
Anaesthesia time3.559 × 10−50.0000−0.0180.00182.5910.2658−37.188710.973347.0131.74 × 10−2216.32
Recovery time2.737 × 10−40.0000−0.1360.023017.0333.4830522.9480145.107041.0751.35 × 10−20219.44
Table 2. Time (in seconds) required to induce sedation and anaesthesia and recovery time of Macrobrachium rosenbergii post-larvae exposed to different concentrations of Ocimum gratissimum essential oil (EO-OG).
Table 2. Time (in seconds) required to induce sedation and anaesthesia and recovery time of Macrobrachium rosenbergii post-larvae exposed to different concentrations of Ocimum gratissimum essential oil (EO-OG).
EO-OG Concentrations
(μL L−1)		Mean (s)S.E.Min (s)Max (s)
Sedation5044.801.2231.0058.00
10041.531.8025.0063.00
15049.241.1833.0062.00
20015.980.6310.0023.00
30025.851.2914.0047.00
Anaesthesia5054.171.1240.0066.00
10060.432.5732.0095.00
15080.102.9741.00112.00
20022.521.0212.0045.00
30048.161.6830.0080.00
Recovery501006.6025.20649.001296.00
1001367.1019.801116.001554.00
150630.1011.30502.00770.00
200829.6014.40693.001005.00
300765.0015.30600.00948.00
S.E.: standard error. Min: minimum. Max: maximum.
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MDPI and ACS Style

de Souza Valente, C.; Mendes, A.F.; do Nascimento Ferreira, C.H.; Baldisserotto, B.; Heinzmann, B.M.; Vaz-dos-Santos, A.M.; Ballester, E.L.C. Anaesthetic Effect of Clove Basil (Ocimum gratissimum) Essential Oil on Macrobrachium rosenbergii Post-Larvae. Aquac. J. 2024, 4, 192-202. https://doi.org/10.3390/aquacj4030014

AMA Style

de Souza Valente C, Mendes AF, do Nascimento Ferreira CH, Baldisserotto B, Heinzmann BM, Vaz-dos-Santos AM, Ballester ELC. Anaesthetic Effect of Clove Basil (Ocimum gratissimum) Essential Oil on Macrobrachium rosenbergii Post-Larvae. Aquaculture Journal. 2024; 4(3):192-202. https://doi.org/10.3390/aquacj4030014

Chicago/Turabian Style

de Souza Valente, Cecília, Arielly Fávaro Mendes, Caio Henrique do Nascimento Ferreira, Bernardo Baldisserotto, Berta Maria Heinzmann, André Martins Vaz-dos-Santos, and Eduardo Luis Cupertino Ballester. 2024. "Anaesthetic Effect of Clove Basil (Ocimum gratissimum) Essential Oil on Macrobrachium rosenbergii Post-Larvae" Aquaculture Journal 4, no. 3: 192-202. https://doi.org/10.3390/aquacj4030014

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