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Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii



Wan Noazira Wan Adnan, Nor Juneta Abu Seman, Nurul Ulfah Karim, Safiah Jasmani, Nor Asma Husna Yusoff and Marina Hassan
 
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ABSTRACT

Background and Objective: Studies on plant herbs as alternatives to chemical anaesthetics in fish species are numerous, but little is known on crustaceans. A study was conducted to investigate the efficacy of C. citratus Essential Oil (EO) on the induction and recovery of M. rosenbergii. Materials and Methods: The C. citratus EO was obtained by hydrodistillation and analyzed using GC-MS. The prawns were exposed to C. citratus EO and clove oil in 100-1000 and 200-1000 μL L–1, respectively. Different stages of induction and recovery times were recorded. Results: In GC-MS, citral (78.47%) was detected as a major compound in C. citratus EO. Prawns reached loss equilibrium at 500-1000 μL L–1 C. citratus EO within 15.55-6.52 min. Exposure of prawn to <500 μL L–1 C. citratus EO resulted in a high survival rate (100-94%). In clove oil, all tested concentrations caused significant induction in M. rosenbergii within 20.61-6.47 min. Recovery time and survival rate were significantly decreased with the increase of EO concentrations. The regression model showed the induction time in both anaesthetic agents was dependent on the concentration (R2 = 0.86-0.96). The recovery time of C. citratus EO-exposed prawn was dependent on the concentrations (R2 = 0.59). Conclusion: The study shows the potentiality of C. citratus EO as a natural anaesthetic in M. rosenbergii, although not as efficient as clove oil.

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  How to cite this article:

Wan Noazira Wan Adnan, Nor Juneta Abu Seman, Nurul Ulfah Karim, Safiah Jasmani, Nor Asma Husna Yusoff and Marina Hassan, 2021. Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii. Pakistan Journal of Biological Sciences, 24: 756-764.

DOI: 10.3923/pjbs.2021.756.764

URL: https://scialert.net/abstract/?doi=pjbs.2021.756.764
 
Copyright: © 2021. This is an open access article distributed under the terms of the creative commons attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited.

INTRODUCTION

Anaesthetics are known to be effective in minimizing physical injuries and stress in prawns particularly during handling and transportation1. Tricaine methanesulfonate (MS-222) is a commonly used anaesthetic for aquatic organisms2. However, its usage is reported could cause retinopathy to the users, besides has a certain withdrawal period before product consumption2. Furthermore, this MS-222 also is not effective in many crustacean species3. Recently, the usage of natural plants as an anaesthetic in the aquaculture sector has attracted a lot of attention among researchers. Plant extracts such as Essential Oils (EO) and other active compounds are being reported to be more effective, less expensive and safer for aquatic life. Chilled sawdust has been reported efficiently in anaesthetizing M. rosenbergii during transportation4. A combination of eugenol and menthol effectively anaesthetize adult M. rosenbergii during the handling process1.

Cymbopogon citratus is a plant from the family Gramineae and is widely cultivated in temperate and tropical regions including Indochina, Indonesia and Malaysia5. It is commonly known as lemongrass and popular in Malay, Thai and Vietnamese cuisine; frequently used in food processing as a food flavouring, perfume and cosmetic industry5. Recent studies have demonstrated that essential oil from the genus Cymbopogon has sedative effects on Silver Catfish (Rhamdia quelen) and Tambaqui (Colossoma macropomum)6,7. However, to date, there is still no information on the anaesthetic/sedative properties of C. citratus EO on crustaceans. As citral oil can produce anxiolytic, sedative and motor relaxant effects in mice8, it was then hypothesized herein that C. citratus EO in appropriate concentrations, would promote an anaesthetic effect on giant freshwater prawn (Macrobrachium rosenbergii) as well.

Therefore, the present study aimed to determine the anaesthetic potential of the C. citratus EO through the evaluation of M. rosenbergii subjected to different concentrations of C. citratus EO and their recovery times. The survival rate after anaesthesia also was assessed.

MATERIALS AND METHODS

Study area: The study was carried out in the Institute of Aquaculture Tropical and Fisheries, Universiti Malaysia Terengganu (UMT), Terengganu, Malaysia from March, 2013-September, 2017.

Preparation of experimental animals: Macrobrachium rosenbergii with an average body weight of 18-23 g were purchased from a local farmer at Jelebu, Negeri Sembilan and transported to the Institute of Tropical Aquaculture and Fisheries (AKUATROP) hatchery for acclimation. Animals were maintained in a continuously aerated fibreglass tank (size: 0.3×0.05×0.1 m) with controlled water and oxygen parameters; water temperature: 27±1°C, pH7.6±0.5, dissolved oxygen: 8.5±0.7 mg L–1 and total ammonia level: 0.5±0.5 mg L–1. Dissolved oxygen was measured by YSI oxygen meter (Model Y556, USA) while a total ammonia level in the water was determined using the API Ammonia Test Kit (Mars® Fishcare) (MARS Fishcare, USA). Acclimatization was conducted for 7 days in a Semi-static system where half of the water volume in the tank was changed daily. Prawns were fed twice daily with commercial pellet and starved for 24 hrs before the experiments. Thirty healthy prawns were used in every treatment (n = 30), with ten per replicate and each treatment consisted of three replicates.

Plant collection: Fresh leaves of Cymbopogon citratus were collected from the local market at Gong Badak and Wakaf Tengah, Kuala Terengganu from September, 2012-March, 2013. Samples were cut and dried for 3-5 days in a ventilated drying oven (Memmert Model: SN30, Germany) at 25°C. Dried samples were ground into fine powder form by using a heavy-duty Waring blender (Fisher Scientific, USA) and kept in a sealed dark plastic bag at room temperature (27°C) until extraction started.

Plant extraction and analysis: The Essential Oil (EO) of C. citratus was extracted by hydrodistillation method using a Clevenger type apparatus for 3 hrs9. Collected EO yields were calculated w/w (%) before stored in amber glass bottles at -4°C until further analysis. Analysis of the EO was performed by Hewlett Packard 6890 Gas Chromatography coupled (GC) with Mass Spectrophotometer (MS) (Agilent Technologies, USA) equipped with DB-5 silica capillary column (Agilent Technologies, USA); (size: 30 m×0.25 mm×0.25 μm). The conditions were as follows: detector and injector temperature, 240°C; carrier gas, helium (99.99% purity) at 1.7 mL min–1. The identification of the EO constituents was made through the comparison of substances mass spectrum with the database of the GC-MS (NIST. lib) (Agilent Technologies, USA), literature and retention time index10.

Table 1: Induction stages and behaviour observation adopted by Vartak and Singh11
Image for - Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii

Anaesthesia induction and recovery: Clove oil and C. citratus EO were used in this experiment. Tested concentrations were as follow: clove oil (200, 300, 400, 500, 600, 700, 800, 900 and 1000 μL L–1); C. citratus EO (100, 200, 300, 400, 500, 600,700, 800, 900 and 1000 μL L–1). Firstly, the anaesthetic agent was dissolved in ethanol at a ratio of 1:10. The control treatment used solely ethanol following the highest concentration of anaesthetic agent. M. rosenbergii were caught using a dip net and introduced individually into the anaesthetic bath (size: 35×20×24 cm) containing 10 L of the treatment concentration. Once the prawns reached the last stage of anaesthesia (total loss equilibrium), they were then transferred to a tank with clean water to observe for recovery for a maximum of 30 min and the full recovery time was recorded. After recovery, prawns were grouped according to treatments and transferred to continuously-aerated 50 L tanks and reared for 24 hrs. Any mortality within these 24 hrs was recorded. The behaviours of M. rosenbergii were characterized according to the criteria evaluation of the stages of induction and recovery by Vartak and Singh11, with slight modifications (Table 1). The time required for inducing anaesthesia and recovery was monitored. Similar methods and data observations were applied in both anaesthetics’ agents.

Statistical analysis: All data were expressed as Mean±SEM. For anaesthesia induction and recovery, data normality was assessed by the Shapiro-Wilk test and the non-normal data was submitted to a square root transformation and analyzed using a one-way analysis of variance (ANOVA) followed by Tukey’s tests. Analysis was performed with MINITAB 16.0 software and p<0.05 was considered statistically significant.

RESULTS

Yield and GC/MS profiling for C. citratus EO: The hydrodistillation process of 880 g C. citratus yielded 5.61 g of EO which equally to 0.72% (w/w) of the total yield percentage. Forty-two volatile components were identified in C. citratus EO. Citral was found to be a major compound (78.47%), followed by 5-Hepten-1-yne-methyl (3.83%), β-myrcene (3.34%), β-ocimene (1.2%), Ethenyl-cyclohexane (1.63%) and Selina-6-en-4-ol (1.38%) (Table 2).

Short-term anaesthesia and recovery: Induction and recovery time of prawn tested with C. citratus EO and clove oil are presented in Table 3 and 4, respectively. Both anaesthetic agents tested in this study showed sedative and anaesthetic effects in M. rosenbergii. The application of sole ethanol did not produce any anaesthetic effect. Prawn exposed to C. citratus EO started to experience Light Sedation (LS) stage ranging from 300-400 μL L–1 after 20.68-12.56 min of exposure time. However, this concentration was unable to promote prawn into subsequent induction stages even after 30 min of observations. Increased of C. citratus EO to 500 μL L–1 led to 5.38 min of LS stage with totally loss equilibrium (deep anaesthetic) at 15.55 min and the subsequent increasing concentration reduced the induction time. The fastest induction was at 1000 μL L–1 with LS at 2.01 min and loss equilibrium at 6.52 min. The overall trend shows that the induction time was significantly dependent on the concentration of C. citratus EO (R2 = 0.86-0.93) (Fig. 1). The reduction of induction time occurred with the increase of C. citratus EO concentration. Meanwhile, the opposite trend was identified in recovery time where the increase of C. citratus EO concentrations resulted in an elevation of the recovery time (Fig. 2). No mortality was observed within the range of 100-300 μL L–1 of C. citratus EO exposure. A 94% of survival rate was found at 500 μL L–1 of C. citratus EO treatment after 24 hrs of observations.

In clove oil, prawn exposed to all concentrations exhibited sedation and anaesthetic effects started at the lowest concentrations; 200 μL L–1 with 20.61 min to LS stage and 29.25 min to reach into totally loss equilibrium (deep anaesthetic) stage. Generally, clove oil concentrations significantly affected the induction and recovery time (p<0.05) in M. rosenbergii (Fig. 3-4). The increased concentrations of clove oil significantly decreased the time needed for sedation and anaesthetic (R2 = 0.91-0.96).

Table 2: Chemical compositions of essential oil from C. citratus by GC-MS analysis
Image for - Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii

Table 3: Required time for the stages of induction and recovery from anaesthesia at different concentrations of C. citratus EO in M. rosenbergii
Image for - Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii
LS: Light sedation, DS: Deep sedation, PLE: Partial loss equilibrium, TLE: Total loss equilibrium. Values are expressed as Mean±SD. Different lowercase superscripts in the same column indicate statistical difference among concentrations and different uppercase in the same row indicate statistical difference among stages after ANOVA and Tukey test (n = 30, p<0.05)

Image for - Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii
Fig. 1:Relationship between stages of induction time with C. citratus EO concentrations

Image for - Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii
Fig. 2:Relationship of recovery time with C. citratus EO concentrations

Table 4: Required time for the stages of induction and recovery from anaesthesia at different concentrations of clove oil in M. rosenbergii
Image for - Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii
LS: Light sedation, DS: Deep sedation, PLE: Partial loss equilibrium, TLE: Total loss equilibrium. Values are expressed as Mean±SD. Different lowercase superscripts in the same column indicate statistical difference among concentrations and different uppercase in the same row indicate statistical difference among stages after ANOVA and Tukey test (n = 30, p<0.05)

Image for - Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii
Fig. 3:Relationship between stages of induction time with clove oil concentrations

Image for - Anaesthetic Efficiency of Cymbopogon citratus Essential Oil and Clove Oil on Macrobrachium rosenbergii
Fig. 4:Relationship of recovery time with clove oil concentrations

However, clove oil survival rate was contradicted with the C. citratus EO trend, wherein clove oil treatment the trend was increased with the increase of clove oil concentrations. The highest survival rate (92%) was observed at the highest treatment of clove oil (1000 μL L–1).

Modelling of induction time versus anaesthetic concentration showed that induction time was strongly dependent on the anaesthetic concentrations in all stages of induction in both C. citratus (Fig. 1) and clove oil (Fig. 3); R2 = 0.86-0.93, R2 = 0.91-0.96, respectively. Meanwhile, modelling of recovery time versus concentrations in C. citratus EO (Fig. 2) can be used in predicting the recovery times in M. rosenbergii (R2 = 0.59). However, clove oil recovery times does not dependent on the concentration of clove oil (R2 = 0.7977) (Fig. 4).

DISCUSSION

In this present study, hydrodistillation of C. citratus yielded a total of 0.715% which is following Viana et al.12, but contradicted with Jalal et al.13. Differences might be due to the different techniques applied in sample preparations, extraction methods and sample preservations as volatile compounds in plant cells are highly sensitive and easy to get release into the environment9. Meanwhile, citral was detected as a major compound in C. citratus EO in the percentage of 78.47%, followed by 5-hepten-yne-6 dimethyl (3.38%), β-myrcene (3.34%), ethnyl cyclohexane (1.63%), selina 6-en-4-ol (1.38%) and β-trans-ocimene (1.23%) (Table 1). The finding was similar to the previous study where they also found citral as a major compound in C. citratus EO but differs in percentage amounts14-17. Meanwhile, myrcene and geraniol were reported to be dominant in other Cymbopogon species instead of citral7,18. Variations in extract compositions are influenced through several factors including geographical area, harvesting period and extraction methods19,20. Citral contained EO have documented attributes to various biological activities including antimicrobial, antifungal, antioxidant, anti-depressant, anti-inflammatory, sedation and/or anaesthetic effects7,12,20-24.

The data of Table 3 and 4 represent the induction time, recovery and survival of both anaesthetic agents; C. citratus EO and clove oil in M. rosenbergii, as respectively. C. citratus EO as low as 200-300 μL L–1 induced M. rosenbergii to light sedation stage but no observation of deep anaesthesia even after 30 min of exposure. Although a low concentration of C. citratus EO did not determine deep anaesthesia, it is capable to promote sedation effect on M. rosenbergii with the shortest induction time at 12.56 min, 400 μL L–1. This finding was in similar range with C. nardus EO for sedation effect in tambaqui fish7. However, a similar concentration of menthol (200 μL L–1) promoted the latest sedation effect in adult M. rosenbergii, 202.3 min. This differential efficiency might be due to the chemical specificity of receptors1. Meanwhile, dos Santos et al.6 also reported the sedation effect of silver catfish at a low concentration of C. flexuosus EO (25 μL L–1). This sedation effect is important particularly during non-invasive procedures such as tagging, biometrics and transportation25. A total loss equilibrium/deep anaesthesia sometimes may not be desirable during transport, as overcrowding of sedated prawn at the bottom tank might lead to asphyxiation26. Besides, prawn with highly sedated may experience mechanical injury by hitting the tank walls during transportation27.

Plotted graph (Fig. 1) shows a clear concentration-dependent trend in C. citratus EO towards the induction of M. rosenbergii to anaesthesia. The optimum concentration which exhibited rapid sedation and/or deep anaesthesia with fast recovery and high survival rate was at 500 μL L–1. Even though the time for induction was exceeded the recommended standards of induction; 3-5 min28, but the time for recovery was rapid (<10 min), with a high survival rate (94%), which met the standards criteria for commercial anaesthetics. This rapid recovery was presumably because of the fast release of drug uptakes via the gills7.

In clove oil treatment, the result shows the ability of cove oil to cause total loss equilibrium (deep anaesthesia) in M. rosenbergii at 1000 μL L–1 within 6.47 min. Meanwhile, the light sedation stage was started even earlier at 200 μL L–1 after 20.61 min treatment. The induction time of clove oil in this study was shorter than reported by Vartak and Singh11 in the same species of prawn. Differences might be due to the experimental conditions such as water prawn size, disease status, stress and other physicochemical properties at the moment of induction29,30. As reported by other previous research; fish weight is significantly influenced by the time needed for the induction31.

Results showed that all stages in induction time are strongly predictable using concentrations of C. citratus EO and clove oil. However, recovery time in the clove oil exposed prawn is increased with the increasing of concentrations. The regression model is important for the determination of the interrelation of y-variables with -variables. However, further study on the respective compound which is involved in this anaesthetic/sedation effect might be an advantage if could be determined, which now become our limitation in this study. Hence, the treatment of the isolated single compounds in M. rosenbergii might be important for a thorough determination of the mechanisms occurring during anaesthesia in crustaceans.

CONCLUSION

In conclusion, C. citratus EO can be used for prawn anaesthesia, although it is not as efficacious as clove oil, the most commonly-used herbal anaesthetic. C. citratus EO at 300-400 μL L–1 can be used safely for inducing sedation in M. rosenbergii. These sedation effects are very useful, particularly during non-invasive procedures. Meanwhile, 500 μL L–1 of C. citratus EO is suitable in promoting the anaesthesia effect (5.38-15.55 min) with fast recovery (<10 min) and a high survival rate (92%). Thus, it can be recommended as a new potential plant anaesthetic in M. rosenbergii species.

SIGNIFICANCE STATEMENT

This study discovers the relationship between induction and recovery time at a varying concentration of C. citratus and clove oil in M. rosenbergii that can be beneficial for comparison between the newly found natural aesthetic with the current-accepted-anaesthetic clove oil. This study will help the researchers out there particularly people in aquatic fields to uncover the potentiality of C. citratus EO as an anaesthetic agent in freshwater prawns which is still rarely studied compared to fish. Thus, a new theory on the dose requirement for anaesthetic/sedative effect on M. rosenbergii may be arrived at.

ACKNOWLEDGMENTS

This research was funded by the Higher Institution Centre of Excellence (HICoE), Institute of Aquaculture Tropical and Fisheries (AKUATROP), UMT under the vote number of 66955. The author also would like to pay our respects to Dr. Safiah Jasmani who had passed away in September, 2017, for her contributions in initiating and planning this study.

REFERENCES

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

2:  Popovic, N.T., I. Strunjak‐Perovic, R. Coz‐Rakovac, J. Barisic, M. Jadan, A.P. Berakovic and R.S. Klobucar, 2012. Tricaine methane-sulfonate (MS-222) application in fish anaesthesia. J. Applied Ichthyol., 28: 553-564.
CrossRef  |  Direct Link  |  

3:  Coyle, S.D., R.M. Durborow and J.H. Tidwell, 2004. Anesthetics in Aquaculture. SRAC Publication, Texas, pp: 6
Direct Link  |  

4:  Salin, K.R., 2005. Live transportation of Macrobrachium rosenbergii (De Man) in chilled sawdust. Aquac. Res., 36: 300-310.
CrossRef  |  Direct Link  |  

5:  dos Santos, A.C., G.B. Junior, D.C. Zago, C.C. Zeppenfeld and D.T. da Silva et al., 2017. Anesthesia and anesthetic action mechanism of essential oils of Aloysia triphylla and Cymbopogon flexuosus in silver catfish (Rhamdia quelen). Vet. Anaesth. Analg., 44: 106-113.
CrossRef  |  Direct Link  |  

6:  Barbas, L.A.L., M. Hamoy, V.J. de Mello, R.P.M. Barbosa and H.S.T. de Lima et al., 2017. Essential oil of citronella modulates electrophysiological responses in tambaqui Colossoma macropomum: A new anaesthetic for use in fish. Aquaculture, 479: 60-68.
CrossRef  |  Direct Link  |  

7:  do Vale, T.G., E.C. Furtado, J.G. Santos and G.S.B. Viana, 2002. Central effects of citral, mycrene and limonene, constituents of essential oil chemotypes from Lippia alba (Mill.) N.E. Brown. Phytomedicine, 9: 709-714.
CrossRef  |  Direct Link  |  

8:  Kulkarni, S.B., M.Y. Kariduraganavar and T.M. Aminabhavi, 2003. Molecular migration of aromatic liquids into a commercial fluoroelastomeric membrane at 30, 40 and 50°C. J. Appl. Polym. Sci., 90: 3100-3106.
CrossRef  |  Direct Link  |  

9:  Kasali, A.A., A.O. Oyedeji and A.O. Ashilokun, 2001. Volatile leaf oil constituents of Cymbopogon citratus (DC) Stapf. Flavour Fragrance J., 16: 377-378.
CrossRef  |  Direct Link  |  

10:  da Cunha, M.A., F.M.C. de Barros, L. de Oliveira Garcia, A.P. de Lima Veeck and B.M. Heinzmann et al., 2010. Essential oil of Lippia alba: A new anesthetic for silver catfish, Rhamdia quelen. Aquaculture, 306: 403-406.
CrossRef  |  Direct Link  |  

11:  Vartak, V. and R.K. Singh, 2006. Anesthetic effects of clove oil during handling and transportation of the freshwater prawn, Macrobrachium rosenbergii (de man). Isr. J. Aquac., 58: 46-54.
Direct Link  |  

12:  Viana, G.S.B., T.G. Vale, R.S.N. Pinho and F.J.A. Matos, 2000. Antinociceptive effect of the essential oil from Cymbopogan citratus in mice. J. Ethnopharmacol., 70: 323-327.
CrossRef  |  

13:  Elyemni, M., B. Louaste, I. Nechad, T. Elkamli and A. Bouia et al., 2019. Extraction of essential oils of Rosmarinus officinalis L. by two different methods: Hydrodistillation and microwave assisted hydrodistillation. Scient. World J., Vol. 2019.
CrossRef  |  Direct Link  |  

14:  Nakamura, Y., M. Miyamoto, A. Murakami, H. Ohigashi, T. Osawa and K. Uchida, 2003. A phase II detoxification enzyme inducer from lemongrass: Identification of citral and involvement of electrophilic reaction in the enzyme induction. Biochem. Biophys. Res. Commun., 302: 593-600.
Direct Link  |  

15:  Blanco, M.M., C.A.R.A. Costa, A.O. Freire, J.G. Santos Jr. and M. Costa, 2009. Neurobehavioral effect of essential oil of Cymbopogon citratus in mice. Phytomedicine, 16: 265-270.
CrossRef  |  Direct Link  |  

16:  Aiemsaard, J., S. Aiumlamai, C. Aromdee, S. Taweechaisupapong and W. Khunkitti, 2011. The effect of lemongrass oil and its major components on clinical isolate mastitis pathogens and their mechanisms of action on Staphylococcus aureus DMST 4745. Res. Vet. Sci., 91: e31-e37.
CrossRef  |  Direct Link  |  

17:  Costa, C.A.R.A., D.O. Kohn, V.M. Lima, A.C. Gargano, J.C. Florio and M. Costa, 2011. The GABAergic system contributes to the anxiolytic-like effect of the essential oil from Cymbopogon citratus (lemongrass). J. Ethnopharmacol., 137: 828-836.
CrossRef  |  Direct Link  |  

18:  Tajidin, N.E., S.H. Ahmad, A.B. Rosenani, H. Azimah and M. Munirah, 2012. Chemical composition and citral content in lemongrass (Cymbopogon citratus) essential oil at three maturity stages. Afr. J. Biotechnol., Vol. 11.
CrossRef  |  Direct Link  |  

19:  Tondolo, J.S.M., L.D.P. Amaral, L.N. Simões, Q.I. Garlet and B. Schindler et al., 2013. Anesthesia and transport of fat snook Centropomus parallelus with the essential oil of Nectandra megapotamica (Spreng.) Mez. Neotrop. Ichthyol., 11: 667-674.
CrossRef  |  Direct Link  |  

20:  Wang, Y.S., Z.Q. Wen, B.T. Li, H.B. Zhang and J.H. Yang, 2016. Ethnobotany, phytochemistry, and pharmacology of the genus Litsea: An update. J. Ethnopharmacol., 181: 66-107.
CrossRef  |  Direct Link  |  

21:  Kamle, M., D.K. Mahato, K.E. Lee, V.K. Bajpai, P.R. Gajurel, K.S. Gu and P. Kumar, 2019. Ethnopharmacological properties and medicinal uses of Litsea cubeba. Plants, Vol. 8.
CrossRef  |  Direct Link  |  

22:  Villalobos, M.C., 2015. Antioxidant activity and citral content of different tea preparations of the above-ground parts of lemongrass (Cymbopogon citratus Stapf.). J. Agric. Food Chem., 46: 1111-1115.
Direct Link  |  

23:  Martins, H.B., N. das Neves Selis, C.L. Silva e Souza, F.S. Nascimento and S.P. de Carvalho et al., 2017. Anti-inflammatory activity of the essential oil citral in experimental infection with Staphylococcus aureus in a model air pouch. Evidence-Based Compl. Alt. Med., Vol. 2017.
CrossRef  |  Direct Link  |  

24:  Sneddon, L.U., 2012. Clinical anesthesia and analgesia in fish. J. Exot. Pet Med., 21: 32-43.
CrossRef  |  Direct Link  |  

25:  Coyle, S.D., S. Dasgupta, J.H. Tidwell, T. Beavers, L.A. Bright and D.K. Yasharian, 2005. Comparative efficacy of anaesthetics for the freshwater prawn Macrobrachium rosenbergii. J. World Aquacult. Soc., 36: 282-290.
CrossRef  |  Direct Link  |  

26:  Wagner, G.N., T.D. Singer and R.S. McKinley, 2003. The ability of clove oil and MS-222 to minimize handling stress in rainbow trout (Oncorhynchus mykiss walbaum). Aquac. Res., 34: 1139-1146.
CrossRef  |  Direct Link  |  

27:  Gilderhus, P.A. and L.L. Marking, 1987. Comparative efficacy of 16 anesthetic chemicals on rainbow trout. North Am. J. Fish. Manage., 7: 288-292.
CrossRef  |  Direct Link  |  

28:  Gomes, D.P., B.W. Chaves, A.G. Becker and B. Baldisserotto, 2011. Water parameters affect anaesthesia induced by eugenol in silver catfish, Rhamdia quelen. Aquacult. Res., 42: 878-886.
CrossRef  |  Direct Link  |  

29:  Bowker, J.D., J.T. Trushenski, D.C. Glover, D.G. Carty and N. Wandelear, 2015. Sedative options for fish research: A brief review with new data on sedation of warm-, cool-, and coldwater fishes and recommendations for the drug approval process. Rev. Fish Biol. Fish., 25: 147-163.
CrossRef  |  Direct Link  |  

30:  Tsantilas, H., A.D. Galatos, F. Athanassopoulou, N.N. Prassinos and K. Kousoulaki, 2006. Efficacy of 2-phenoxyethanol as an anaesthetic for two size classes of white sea bream, Diplodus sargus L. and sharp snout sea bream, Diplodus puntazzo C. Aquaculture, 253: 64-70.
CrossRef  |  Direct Link  |  

31:  Adnan, W.N.W., N.U. Karim, N.A.H. Yusoff, M.I. Zakariah and M. Hassan, 2021. Effect of Cymbopogon citratus essential oil (EO) on handling stress in giant freshwater prawn (Macrobrachium rosenbergii). Pak. J. Bio. Sci., 24: 13-18.
CrossRef  |  Direct Link  |  

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