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Article

Dietary Ocimum gratissimum Essential Oil Improves the Antioxidant Status and Maintains the Performance of Macrobrachium rosenbergii Juveniles

by
Eduardo Luis Cupertino Ballester
1,*,
Wilson Gilberto da Silva Costa
1,
Caio Henrique do Nascimento Ferreira
2,
Milena Cia Retcheski
3,
Luisa Helena Cazarolli
3,
Gabriel Henrique Schwengber
4,
Marlise Teresinha Mauerwerk
1,
Carlos Garrido Pinheiro
5,
Berta Maria Heinzmann
5,6,
Bernardo Baldisserotto
7 and
Cecília de Souza Valente
8
1
Shrimp Culture Laboratory, Post Graduation Program in Aquaculture and Sustainable Development, Federal University of Paraná, Palotina 85953-128, Parana, Brazil
2
Post Graduation Program in Aquaculture, Faculty of Agricultural and Veterinary Sciences, São Paulo State University, Sao Paulo 14884-900, Sao Paulo, Brazil
3
Biochemistry and Genetics Laboratory, Federal University of Fronteira Sul—Laranjeiras do Sul Campus, BR 158, Km 405, Rural Zone, Laranjeiras do Sul 85301-970, Parana, Brazil
4
Graduate Program in Aquaculture Engineering, Federal University of Paraná, Palotina 85953-128, Parana, Brazil
5
Graduate Program in Forest Engineering, Federal University of Santa Maria, Santa Maria 97105-900, Rio Grande do Sul, Brazil
6
Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria 97105-900, Rio Grande do Sul, Brazil
7
Department of Physiology and Pharmacology, Federal University of Santa Maria, Santa Maria 97105-900, Rio Grande do Sul, Brazil
8
School of Biological and Chemical Sciences, University of Galway, H91 TK33 Galway City, Ireland
*
Author to whom correspondence should be addressed.
Appl. Sci. 2025, 15(5), 2745; https://doi.org/10.3390/app15052745
Submission received: 21 January 2025 / Revised: 23 February 2025 / Accepted: 2 March 2025 / Published: 4 March 2025
(This article belongs to the Special Issue Advances in Aquatic Animal Nutrition and Aquaculture)
Versions Notes

Abstract

:
This study analyzed the performance, antioxidant status, hepatopancreatic lipoperoxidation, and proximate composition of Macrobrachium rosenbergii juveniles fed diets supplemented with clove basil (Ocimum gratissimum) essential oil (EO-OG). A total of 360 M. rosenbergii (initial weight 0.028 g ± 0.004) were randomly divided into four experimental groups with six replications each (n = 6). The prawns were fed diets with different EO-OG inclusion levels: 0.0, 1.0, 2.0, and 3.0 g kg−1 EO-OG. After a 42-day feeding trial, dietary EO-OG showed no significant effect on prawn performance or carcass proximate composition, except on final antenna length. Prawns fed 3.0 g kg−1 EO-OG displayed a 1.2- to 1.3-fold longer final antenna length than prawns from all other experimental groups. Likewise, prawns fed 3.0 g kg−1 EO-OG presented a 2.6- to 3.2-fold higher catalase activity than prawns from all other experimental groups. Prawns fed EO-OG, regardless of the inclusion level, showed a 1.6- to 1.7-fold decreased hepatopancreatic lipoperoxidation compared to the control group. Therefore, EO-OG has been demonstrated to be a potential management tool as a non-nutritional dietary immunostimulant and animal welfare promoter for freshwater prawn farming, without affecting animal performance. This study recommends the dietary inclusion level of 3.0 g kg−1 EO-OG for M. rosenbergii juveniles.

1. Introduction

Aquaculture plays a key role in food security and nutrition for the world’s population and it provides some of the main edible protein sources. It supplies the global market with diverse aquatic species raised in different production systems that suit local activity conditions. Among them, farming of the giant river prawn (Macrobrachium rosenbergii) has gained prominence due to its resistance to disease and high market value. In 2020, the M. rosenbergii global production reached 294,000 tons, the equivalent of 2.6% of total crustacean farming [1].
Aiming at increasing productivity and economic gains and promoting animal health in crustacean farming, studies have been conducted to develop effective growth promoters and antioxidant and immunostimulant compounds. They include probiotics [2], prebiotics [3], organic acids [4], plant extracts [5,6], and essential oils [7,8]. From this perspective, the use of natural compounds has increased in recent years, particularly the use of essential oils (EOs). The EOs are volatile, odorous, and lipophilic substances extracted from the secretory glands of a plant, such as the leaves, flowers, roots, and seeds. They are abundant in low-molecular-weight mono- and sesquiterpenoids that provide the unique biological properties of EOs [9,10].
The biological activities of EOs used in aquaculture include antioxidant [11,12,13], antiparasitic and antimicrobial [12,13,14], immunostimulant [15,16], and anti-inflammatory properties [17,18], as well as growth promotion [7,13], anesthetic [13,19,20], gut health [21], and food preservative [22,23] activities. In particular, recent studies have investigated the influence of different EOs on decapods performance. Some dietary EOs have shown no effect on animals’ growth or survival, including the EOs of Aloysia triphylla [11] and Lippia alba [24,25] in M. rosenbergii, and a blend of organic acids (citric acid and sorbic acid) and EOs (thymol and vanillin) in Penaeus vannamei [26]. Meanwhile, Cuminum cyminum (5 and 10 g kg feed−1) improved the performance of Penaeus vannamei [27].
The plant Ocimum gratissimum, commonly known as clove basil or African basil, is a tropical perennial aromatic plant in the Lamiaceae family, and it is found in Africa, Asia, and South America [28,29]. Its use encompasses herbal medicine (inc. described properties as an anxiolytic, antimicrobial, anti-diabetic, anti-hypertensive, and antinociceptive agent) and food condiment [30]. The EO of O. gratissimum (EO-OG) is mostly obtained by steam distillation of the inflorescence and leaves. In the aquaculture context, the use of EO-OG has demonstrated multiple benefits. Its use as a sedative, anesthetic, antimicrobial, antiparasitic, and immunostimulant in fish is well known [31,32,33,34]. In decapods, the use of EO-OG is mostly associated with its sedative and anesthetic properties [20,35,36]. Different chemotypes of EO-OG have been described, including citral, ethyl cinnamate, eugenol, linalool, and thymol [37]. In Brazil, the eugenol EO-OG chemotype is the common one found and the biological activities of this EO are attributed to its major constituent (i.e., eugenol).
The antioxidant defense system is a conserved biochemical defense mechanism in vertebrates and invertebrates. Excess reactive oxygen species (ROS) cause an imbalance in the antioxidant system, causing so-called oxidative stress [38]. The antioxidant system minimizes the negative oxidizing effects of ROS through antioxidant enzymes such as catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), and glutathione-S-transferase (GST; a detoxification enzyme from metabolism phase II), as well as non-enzymatic antioxidants, including reduced glutathione (GSH). Essential oils have antioxidant activity, and their dietary use has been beneficial to farmed shrimp [11,24].
Notably, the use of natural substances such as essential oils is expected to be promoted in aquaculture settings since concerns about sustainability, the overuse of chemicals, and animal health and welfare are now prominent. The novelty of this study lies in the investigation of a sustainable essential oil (O. gratissimum) with the potential use as a beneficial husbandry tool and health promoter for farmed prawns. Nevertheless, despite the known properties of EO-OG in aquaculture, its potential dietary use to promote prawn growth and enhance their antioxidant status has not yet been studied. Therefore, this research aimed to study the influence of different dietary inclusion levels of EO-OG on the performance, antioxidant status, and body proximate composition of M. rosenbergii juveniles.

2. Material and Methods

2.1. Plant Collection and Essential Oil Extraction and Characterization

Leaves of O. gratissimum were acquired from a crop in Frederico Westphalen, at the Federal University of Santa Maria (UFSM) campus, Rio Grande do Sul, Brazil. The specimen was identified by Adelino Alvares Filho and deposited in the herbarium of the Department of Biology (UFSM), registration number SMDB 11167. The essential oil of O. gratissimum (EO-OG) was extracted by steam distillation using a Clevenger-type apparatus for 2 h [39]. The hydrolate and EO-OG were dissociated using a separating funnel through liquid–liquid partition with hexane (PA). The hexane fraction was concentrated in a rotary evaporator at 40 °C to obtain the pure EO-OG. Extracted EO-OG was kept at −4 °C, in an amber glass bottle, until posterior analysis and use. The composition analysis and identification of the main compounds of EO-OG were performed as described in detail by de Souza Valente et al. [20]. Major identified components were eugenol (88.46%), β-caryophyllene (5.30%), copaene (0.80%), E-β-ocimene (0.64%), and germacrene D (0.47%).

2.2. Experimental Diets

Experimental diets had different EO-OG inclusion levels: 0.0, 1.0, 2.0, and 3.0 g kg−1 (Table 1). The chosen dietary levels were based on previous similar studies and former studies of our research group [11,24,40,41]. These studies show that these inclusion levels are recommended to achieve potential beneficial effects (e.g., antioxidant effect). Additionally, due to the under-researched biotoxicity of EO-OG on M. rosenbergii, this study adopted a conservative approach by testing small initial inclusion levels within a small range.
Diets were formulated to have a 30% crude protein inclusion level, using SuperCrac® 2.0 software and following the recommendations of the NRC [42]. For feed preparation, ingredients were graded (0.8 mm particle size) and sieved (250 μm mesh). Subsequently, ingredients were dosed according to the respective diet formulation and mixed for processing. Feed pellets were then moistened with water (35 °C), extruded into 1 mm diameter pellets, and dried at 45 °C for 24 h. Final feed pellets were stored at 4 °C until further use.

2.3. Experimental Design

A total of 360 M. rosenbergii juveniles (initial weight 0.028 ± 0.004 g, mixed sex, male–female ratio 50:50%) were randomly allocated into 24 tanks (experimental units), with a 50 L capacity. Each tank had an individual aeration system and was coupled to a recirculation system (flow rate 800 L h−1) with mechanical (Perlon® wool filter; AquaUra, Uberaba, Minas Gerais, Brazil) and biological filters. Prawns were divided into four experimental groups with six replications (n = 6). The stocking density was 75 prawns m−2, equivalent to 15 prawns tank−1. The feeding trial lasted 42 days. The chosen duration of the feeding trial considered that indoor nurseries of M. rosenbergii in tropical regions commonly last from two to eight weeks [43], with the usual time being 30 days. Prawns were fed five times daily (9:00, 11:00, 14:00, 17:00, 21:00) and 30 min after tanks were siphoned to remove feces and uneaten food. The initial feeding rate was equivalent to 5% of the biomass; the feeding rate was adjusted daily according to the feed consumption. Only animals in the intermolt period were used. Mortality was observed twice a day. The natural photoperiod was adjusted for a 12 h light/12 h dark cycle.
Water parameters were within the recommended range for M. rosenbergii farming (mean ± SD): temperature (28.30 ± 0.57 °C), dissolved oxygen (6.45 ± 0.27 mg L−1), pH (8.46 ± 0.10), hardness (124.67 ± 20.77 mg CaCO3 L−1), alkalinity (120.33 ± 10.40 mg CaCO3 L−1), total ammonia nitrogen (0.02 ± 0.04 mg L−1), and nitrite (0.01 ± 0.01 mg L−1). Temperature and dissolved oxygen were determined daily; total ammonia nitrogen and nitrite were monitored weekly; and hardness and alkalinity were measured biweekly.

2.4. Prawn Performance

Survival, weight gain, biomass gain, feed conversion rate, and condition factor were calculated at the end of the feeding trial as indicated by de Souza Valente et al. [11]. Body length was considered as the linear distance from the tip of the rostrum to the tip of the telson. The total length of the antenna was also measured.

2.5. Antioxidant Enzymes and Detoxification Activities in the Hepatopancreas

At the end of the experiment, the hepatopancreases of 60 prawns (15 per experimental group; n = 6 replicates) were randomly collected to analyze the activity of enzymes related to the antioxidant defense system and lipid peroxidation. Prawns were euthanized by thermal shock (5 min in iced water) and the body surface was cleaned with ethanol 70% (v/v). The hepatopancreases were collected and immediately stored at −20 °C until further analysis. Samples were homogenized in 50 mM Tris/HCl buffer pH 7.4 using an electric homogenizer (IKA® T10 basic; IKA®, Staufen, Germany). The entire procedure was carried out on ice to preserve the enzymatic activities. The homogenates were then centrifuged in a refrigerated centrifuge (Sigma, St. Louis, MO, USA, 3-16 KL) at 4 °C for 10 min at 12,000× g and the resulting supernatant was collected for further analysis.
The protein content was determined using the Bradford method [44], using bovine albumin as a standard and absorbance ratio at 595 nm. The protein content was used for subsequent calculations of enzymatic activities. The catalase (CAT) activity was determined based on the decomposition of H2O2 into O2 and H2O, with the consumption of 1 µmol H2O2 min−1, at an absorbance of 240 nm [45,46]. The glutathione peroxidase (GPx) activity was determined by measuring NADPH oxidation at an absorbance of 340 nm, on the reduction of cumene hydroperoxide [47,48]. The glutathione reductase (GR) activity was quantified by measuring the oxidation of NADPH at an absorbance of 340 nm, on the reduction of glutathione disulfide to reduced glutathione [49,50]. Reduced glutathione (GSH) levels were determined by the non-protein thiol method using Ellman’s reagent [50,51]. Glutathione S-transferase (GST) activity was quantified by measuring the conjugation of GSH with 1-chloro-2,4-dinitrobenzene [52,53]. Lipid peroxidation was measured based on the reaction of MDA and thiobarbituric acid reactive species (TBARS) [54,55].

2.6. Prawn Body Proximate Composition

At the completion of the feeding trial, 20 prawns (n = 5 prawns per experimental group) were randomly collected and euthanized by immersion in iced water for 5 min. Proximate composition of the prawn carcasses was carried out using AOAC methods [56]. Briefly, dry matter was quantified by heating samples at 105 °C for 24 h. Crude protein was assayed by Kjeldahl’s method for the measurement of total nitrogen after sulphuric digestion. Mineral material (ash) was obtained after incinerating samples at 550 °C for 12 h. The ether extract (crude lipid) was analyzed by the Soxhlet method and petroleum ether (40–60 °C), gravimetrically.

2.7. Statistical Analysis

Data were checked for homogeneity (Levene’s test) and normality (Shapiro–Wilk’s test). Satisfying the assumptions, data were analyzed using a one-way ANOVA followed by the post-hoc Tukey’s test. Data are expressed as the mean ± SD. A p-value < 0.05 was considered statistically significant.

3. Results

3.1. Prawn Performance

No significant differences were observed between the experimental groups in most of the growth parameters analyzed, except for antenna length. Prawns fed EO-OG 3.0 g kg−1 displayed a 1.2- to 1.3-fold longer final antenna length than all other experimental groups. Survival was similar among all of the experimental groups (Table 2).

3.2. Antioxidant Status

The activity of the catalase enzyme was 2.6- to 3.2-fold higher in prawns fed EO-OG 3.0 g kg−1 than in all of the other experimental groups. Malondialdehyde (lipid peroxidation) was 1.6- to 1.7-fold lower in prawns fed EO-OG-supplemented diets, regardless of the inclusion level, compared to the control group. All other analyzed enzymatic activities were similar among the experimental groups (Table 3).

3.3. Prawn Body Proximate Composition

Prawn body proximate composition was similar among all experimental groups for all measured parameters (Table 4).

4. Discussion

The use of medicinal herbs and their essential oils as dietary supplements for farmed aquatic animals has gained increasing attention due to their biological activities. Different essential oils have beneficial properties including analgesic, immunostimulant, growth promotion, and antioxidant bioactivities [8,15]. In particular, EO-OG can be used to control fish parasitic infection, enhance fish antioxidant response, and act as a fish sedative and anesthetic agent [31,32,33,57], as well as a sedative and anesthetic for prawns [20,36]. In the present study, dietary EO-OG has been shown to be a potential natural antioxidant and contribute to prawn welfare.
When adding a new component to the diet of farmed animals, it is paramount that it does not impair the animals’ performance. Although some EOs have been reported as growth promoters within the aquaculture context, e.g., Citrus sinensis (orange) and Origanum vulgare (oregano) [58,59], some others such as the EOs of Aloysia citriodora (A. triphylla) (lemon verbena), Citrus limon (lemon), Lippia alba (lemon balm), and Rosmarinus officinalis (rosemary) have been shown to have no impact on fish or decapods’ performance [11,24,25,60,61]. Those differences in the growth promoter effect of EOs might be attributed to the major compounds that determine the different EO chemotypes, or minor compounds of the different EOs, or even the set of components present in the EO (phytocomplex), as well as the animal species tested and the experimental conditions. In the current study, no significant differences were observed among experimental groups in most of the growth parameters analyzed, except for the antenna length parameter. Prawns’ performance is influenced by their nutrition, environment, and genetics. The last two were kept equal among the experimental groups throughout the trial; hence, these were not variables. Of note, EO-OG is a non-nutritive feed supplement with a potential immunostimulant effect. Thus, although an enhancement in animal performance might occur, the major expected benefit of EO-OG dietary supplementation is health stimulation.
Prawns fed a diet with 3.0 g kg−1 EO-OG showed longer antenna lengths than prawns from all of the other experimental groups. Antennae are among the main chemosensory organs of decapods and are used for social interaction [62,63]. The antenna length is a physical feature that indicates the welfare status of decapods. Measurement of antenna length can be used as a non-invasive method to infer the welfare of farmed decapods. Long and intact antennae are signs of healthy animals in adequate ambient conditions, while damaged or shortened antennae may suggest poor health and a deteriorated welfare status [64,65]. Thus, based on the longer antennal lengths, dietary supplementation with 3.0 g kg−1 EO-OG supported the prawns’ antenna-related welfare. A future study on prawn behavior may assess how EO-OG supplementation benefits antenna-related behaviors such as competitive interactions, social communication, and mating receptivity.
Normal intracellular reactions and the phagocytic process generate oxidizing compounds such as reactive oxygen species (ROS) and reactive nitrogen intermediates (RNIs). Nevertheless, when in excess, these oxidizing compounds are toxic to the organism’s tissues and need to be neutralized by the antioxidant defense system [66]. The antioxidant defense system is divided into enzymatic (inc. enzymes such as SOD, CAT, and GPx) and non-enzymatic, which in turn can be endogenous (e.g., GSH) or exogenous (e.g., acquired from the diet). The enzyme CAT catalyzes hydrogen peroxide (H2O2) into water (H2O) and oxygen (O2), after the enzyme SOD has catalyzed the superoxide anion (O2−) into H2O2, maintaining a balanced cellular redox status [67]. In this study, prawns fed a diet supplemented with 3.0 g kg−1 EO-OG showed increased CAT enzymatic activity, which suggests a potential antioxidant effect of dietary EO-OG. Likewise, the oxidative damage caused by oxidizing compounds leads to lipid peroxidation [68]. Prawns supplemented with EO-OG, regardless of the inclusion level, displayed decreased lipid peroxidation, which is an indication of a greater efficiency of the antioxidant system of prawns fed with EO-OG. Addressed together, the increased CAT activity may indicate an indirect antioxidant effect of EO-OG that facilitated the decrease in organic peroxides, observed by the diminished lipoperoxidation.
The immune system of decapods relies on innate mechanisms and is mediated by defense cells, physical barriers, and humoral responses. These innate defense mechanisms are intrinsically related to animal health and welfare [69,70,71]. The antioxidant system has a fundamental role in the deactivation of oxidizing compounds formed naturally by physiological processes. Therefore, dietary immunostimulants may enhance decapods’ non-specific defense responses. Notably, the confinement condition of the production systems is a source of chronic stress which can impair the health and welfare of farmed animals, including shrimp and prawns [72,73,74]. Therefore, the use of EO-OG as a potential natural antioxidant can be considered a beneficial management tool to enhance prawns’ health.
Typically, the antioxidant system components (enzymatic and non-enzymatic) operate in concert to maintain body homeostasis. Different antioxidants target different ROS, and they exhibit distinct reaction rates and different mechanisms of action. Our findings indicate that the antioxidant effect of the essential oil is not directly related to classical GSH cycling. First, GSH acts directly in the sequestration of ROS, serving as a reducing agent for disulfides, hydroperoxides, and other electrophilic substances. It may also be involved in the recycling of other antioxidants, such as vitamins C and E, thereby providing reducing power to maintain their reduced forms. Second, GSH functions as a cofactor for other antioxidant systems, including Glyoxalase 1 and Glyoxalase 2, glutathione-dependent peroxiredoxins, and glutathione-dependent thiol: disulfide oxidoreductase (glutaredoxins) [75,76,77]. In addition, the significant reduction in lipid peroxidation can be explained by the consumption of GSH, but also by a direct effect of the phenolic components of the EO, as they can act as reducing agents, hydrogen atom donors, and singlet oxygen scavengers. The radical-scavenging activity of phenolics depends on structural features that favor phenolic hydrogen donation and the stability of the resulting phenoxyl radicals. For example, eugenol (4-allyl-2-methoxyphenol) (the major constituent of EO-OG in this study) is a phenolic compound of the phenylpropanoid class that has a hydroxyl group attached to an aromatic ring in its structure [78,79]. The phenolic hydroxyl and allylic moiety present in eugenol effectively contribute to the antioxidant activity of this natural product. In particular, the unsaturation is very valuable for eugenol’s activity concerning free radicals, since the molecule can stabilize after scavenging the radical on this aromatic ring [80,81].
The antioxidant properties of EO-OG here observed may be attributed to its major and minor compounds. Eugenol was the major compound identified in the EO-OG used in the present study. Eugenol’s free-radical-scavenging, anti-lipoperoxidation, and antioxidant activities have been assigned to its high polyphenolic contents [79,80,82]. Different EOs rich in eugenol exhibit these biological activities and are beneficial in the aquaculture setting. They include EO-OG, Cinnamomum zeylanicum (cinnamon), Eugenia caryophyllus (clove), Rosmarinus officinalis (rosemary), Origanum vulgare (oregano), and Thymus vulgaris (thyme), among others [13,83,84,85].
Likewise, minor constituents of EO-OG may have synergically contributed to the bioactivities of this EO. The sesquiterpene β-caryophyllene has presented properties such as antioxidant, antineoplastic, and antimicrobial ones in human cells and pathogens [86], and antioxidant, anti-inflammatory and wound re-epithelialization activities in rats [87]. In rats, β-caryophyllene showed a cardioprotective effect by reducing the heart lipid content [88] and displayed hypolipidemic effects similar to those of the simvastatin drug [89,90]. Similarly, the tricyclic sesquiterpene copaene displayed antioxidant properties in human lymphocytes [91].
Lastly, the use of EO-OG as a dietary supplement, regardless of the inclusion level, did not affect the carcass proximate composition of M. rosenbergii juveniles. The bromatological composition of aquatic animals such as fish and decapods varies according to the species, sex, age, water parameters, and diet profile [92,93,94]. The results of the current study suggest that EO-OG can be used as a dietary antioxidant without influencing the bromatological composition of prawn carcasses. Notably, one may observe a positive effect on the protein content of prawn biomass, although this was not statistically significant in this study. Forthcoming studies may perform a meta-analysis to show combined effects. Similarly, further research may test higher inclusion levels of EO-OG (i.e., >3.0 g kg −1) to determine the precise optimum dose as well as analyze the activity of digestive enzymes. Likewise, future studies may assess the organoleptic properties of prawns fed a diet supplemented with EO-OG and the potential antioxidant effect of isolated compounds (e.g., eugenol) on M. rosenbergii juveniles.

5. Conclusions

In summary, M. rosenbergii juveniles fed non-nutritive EO-OG maintain performance and body proximate composition while presenting longer antenna length and displaying improved antioxidant status, particularly with the 3.0 g kg−1 dietary inclusion level. Dietary use of EO-OG benefits the antioxidant system of M. rosenbergii juveniles and demonstrates to be a potential natural antioxidant and welfare promoter for farmed freshwater prawns.

Author Contributions

Conceptualization: E.L.C.B.; Resources and funding acquisition: E.L.C.B.; Methodology: W.G.d.S.C. and C.H.d.N.F.; Formal analysis: W.G.d.S.C., C.H.d.N.F., M.C.R., L.H.C., G.H.S. and C.G.P.; Writing—original draft preparation: W.G.d.S.C.; Writing—review and editing: W.G.d.S.C., C.d.S.V., M.T.M., B.M.H., B.B. and E.L.C.B.; Supervision: E.L.C.B. and M.T.M. 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. Bernardo Baldisserotto received a research fellowship from CNPq (process No 301816/2022-0).

Institutional Review Board Statement

Ethical review and approval were waived for this study due to the current absence of animal protective legislation that includes decapod crustaceans in Brazil (Law No 11.794, 2008, Brazil). This study did not involve humans or vertebrates.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data is contained within the article.

Conflicts of Interest

This study was part of Wilson Gilberto da Silva Costa’s Master’s dissertation. The authors declare no other conflicts of interest.

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Table 1. Feed formulation and nutrient analysis of the experimental diets with different Ocimum gratissimum essential oil inclusion levels for Macrobrachium rosenbergii juveniles.
Table 1. Feed formulation and nutrient analysis of the experimental diets with different Ocimum gratissimum essential oil inclusion levels for Macrobrachium rosenbergii juveniles.
Ingredients (g kg−1)Experimental Groups (g kg Diet−1)
Control1.02.03.0
Soybean meal400.00400.00400.00400.00
Wheat bran98.1098.1098.1098.10
Wheat flour154.65154.65154.65154.65
Fish meal150.00150.00150.00150.00
Offal meal92.5092.5092.5092.50
Fish oil42.4041.4040.4039.40
Soy lecithin20.0020.0020.0020.00
Limestone15.6515.6515.6515.65
Salt8.008.008.008.00
Vitamin and mineral premix †8.008.008.008.00
Corn starch5.005.005.005.00
DL-methionine2.902.902.902.90
L-lysine1.601.601.601.60
Antifungal1.001.001.001.00
Butylated hydroxytoluene0.200.200.200.20
EO-OG0.001.002.003.00
Total1000.001000.001000.001000.00
Proximate composition
Moisture (%)8.578.928.699.04
Dry matter (%)91.4391.0891.3190.96
Crude protein (%)39.539.1238.2238.54
Ash (%)9.178.999.468.65
Ether extract (%)7.077.156.97.01
NDF (%)29.4229.4528.2728.6
† Vitamin guarantee levels kg−1: 36 mg vitamin B2, 25.5 mg vitamin B6, 45 mg vitamin B12, 1200 mg vitamin C, 1.68 mg biotin, 11.2 mg folic acid, 265 mg inositol, 170 mg nicotinic acid, 67.5 mg pantothenic acid. Mineral guarantee levels kg−1: 0.35 mg cobalt, 13.8 mg copper, 1.3 mg iodine, 65 mg iron, 85 mg manganese, 0.4 mg selenium, 150 mg zinc. EO-OG: essential oil of Ocimum gratissimum. NDF: neutral detergent fiber.
Table 2. Growth performance of Macrobrachium rosenbergii juveniles fed diets with different Ocimum gratissimum essential oil inclusion levels.
Table 2. Growth performance of Macrobrachium rosenbergii juveniles fed diets with different Ocimum gratissimum essential oil inclusion levels.
Growth ParametersExperimental Groups (g EO-OG kg Diet−1)
Control1.02.03.0p-Value
Initial weight (g)0.028 ± 0.0040.028 ± 0.0040.028 ± 0.0040.028 ± 0.004N/A
Final weight (g)0.62 ± 0.220.66 ± 0.210.61 ± 0.190.62 ± 0.170.4419
Weight gain (g)0.58 ± 0.18 0.64 ± 0.210.58 ± 0.190.60 ± 0.180.2380
Initial biomass (g)0.42 ± 0.06 0.42 ± 0.06 0.42 ± 0.06 0.42 ± 0.06 N/A
Final biomass (g)7.93 ± 0.448.29 ± 0.777.75 ± 0.488.02 ± 0.570.4645
Biomass gain (g)7.51 ± 0.447.87 ± 0.777.33 ± 0.487.60 ± 0.570.4645
Final body length (cm)3.59 ± 0.483.66 ± 0.423.59 ± 0.443.60 ± 0.390.4433
Final antenna length (cm)2.76 ± 0.97 B2.89 ± 0.8 B2.66 ± 0.8 B3.37 ± 0.97 A<0.001
FCR (g g−1)1.50 ± 0.081.44 ± 0.101.54 ± 0.101.48 ± 0.110.5419
Condition factor (K)1.31 ± 0.30 1.31 ± 0.151.31 ± 0.181.30 ± 0.230.9260
Survival (%)97.79 ± 3.4596.65 ± 5.6097.76 ± 3.4598.88 ± 2.730.8126
FCR: feed conversion rate. EO-OG: essential oil of Ocimum gratissimum. Different superscript letters in the same row indicate significant differences. Data presented as mean ± SD, n = 6. One-way ANOVA followed by post-hoc Tukey’s test, p < 0.05.
Table 3. Detoxification biomarkers in the hepatopancreases of Macrobrachium rosenbergii juveniles fed diets with different Ocimum gratissimum essential oil inclusion levels.
Table 3. Detoxification biomarkers in the hepatopancreases of Macrobrachium rosenbergii juveniles fed diets with different Ocimum gratissimum essential oil inclusion levels.
Detoxification BiomarkersExperimental Groups (g EO-OG kg Diet−1)
Control1.02.03.0 p-Value
CAT0.06 ± 0.03 B0.06 ± 0.05 B0.08 ± 0.03 B0.21 ± 0.14 A0.0126
GPx0.05 ± 0.020.03 ± 0.010.05 ± 0.020.05 ± 0.010.3980
GR0.01 ± 0.000.00 ± 0.000.011 ± 0.120.02 ± 0.030.1631
GSH23.19 ± 3.5419.44 ± 12.814.93 ± 7.4017.26 ± 16.470.8167
GST0.02 ± 0.020.02 ± 0.010.02 ± 0.020.04 ± 0.020.2705
TBARS2.80 ± 0.34 A1.69 ± 0.84 B1.65 ± 0.39 B1.59 ± 0.63 B0.0111
CAT: catalase (µmol H2O2/min/mg protein). GPx: glutathione peroxidase (µM NADPH/min/ mg protein). GR: glutathione reductase (NADPH/min/ mg protein). GSH: glutathione (µM/mg protein). GST: glutathione S-transferase (nMol tioeter/min/mg protein). TBARS: lipid peroxidation (µM mg protein). Different superscript letters in the same row indicate significant differences. Data presented as mean ± SD, n = 6. One-way ANOVA followed by post-hoc Tukey’s test, p < 0.05.
Table 4. Proximate composition of Macrobrachium rosenbergii juveniles fed diets with different Ocimum gratissimum essential oil inclusion levels, dry matter based.
Table 4. Proximate composition of Macrobrachium rosenbergii juveniles fed diets with different Ocimum gratissimum essential oil inclusion levels, dry matter based.
Proximate CompositionExperimental Groups (g EO-OG kg Diet−1)
Control1.02.03.0 p-Value
Dry matter (%)24.84 ± 0.5124.11 ± 0.2424.15 ± 0.8723.72 ± 0.800.476
Crude protein (%)68.70 ± 0.7471.49 ± 0.6770.11 ± 1.78 73.52 ± 1.600.077
Ash (%)13.63 ± 0.1013.12 ± 0.2113.21 ± 0.0613.45 ± 0.140.062
Ether extract (%)8.51 ± 0.157.86 ± 0.298.14 ± 0.148.15 ± 0.270.168
Data presented as mean ± SD, n = 5. One-way ANOVA followed by post-hoc Tukey’s test, p < 0.05.
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Ballester, E.L.C.; Costa, W.G.d.S.; do Nascimento Ferreira, C.H.; Retcheski, M.C.; Cazarolli, L.H.; Schwengber, G.H.; Mauerwerk, M.T.; Pinheiro, C.G.; Heinzmann, B.M.; Baldisserotto, B.; et al. Dietary Ocimum gratissimum Essential Oil Improves the Antioxidant Status and Maintains the Performance of Macrobrachium rosenbergii Juveniles. Appl. Sci. 2025, 15, 2745. https://doi.org/10.3390/app15052745

AMA Style

Ballester ELC, Costa WGdS, do Nascimento Ferreira CH, Retcheski MC, Cazarolli LH, Schwengber GH, Mauerwerk MT, Pinheiro CG, Heinzmann BM, Baldisserotto B, et al. Dietary Ocimum gratissimum Essential Oil Improves the Antioxidant Status and Maintains the Performance of Macrobrachium rosenbergii Juveniles. Applied Sciences. 2025; 15(5):2745. https://doi.org/10.3390/app15052745

Chicago/Turabian Style

Ballester, Eduardo Luis Cupertino, Wilson Gilberto da Silva Costa, Caio Henrique do Nascimento Ferreira, Milena Cia Retcheski, Luisa Helena Cazarolli, Gabriel Henrique Schwengber, Marlise Teresinha Mauerwerk, Carlos Garrido Pinheiro, Berta Maria Heinzmann, Bernardo Baldisserotto, and et al. 2025. "Dietary Ocimum gratissimum Essential Oil Improves the Antioxidant Status and Maintains the Performance of Macrobrachium rosenbergii Juveniles" Applied Sciences 15, no. 5: 2745. https://doi.org/10.3390/app15052745

APA Style

Ballester, E. L. C., Costa, W. G. d. S., do Nascimento Ferreira, C. H., Retcheski, M. C., Cazarolli, L. H., Schwengber, G. H., Mauerwerk, M. T., Pinheiro, C. G., Heinzmann, B. M., Baldisserotto, B., & de Souza Valente, C. (2025). Dietary Ocimum gratissimum Essential Oil Improves the Antioxidant Status and Maintains the Performance of Macrobrachium rosenbergii Juveniles. Applied Sciences, 15(5), 2745. https://doi.org/10.3390/app15052745

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