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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.

Abstract This study aimed to evaluate the effectiveness of essential oils (EO) of Pilocarpus pennatifolius (PPOL) and Cordia verbenacea (CVOL) as sedatives and/or anesthetics in juvenile Nile tilapia (Oreochromis niloticus). The main constituents of PPOL were 2-undecanone (57.2%) and 2-Tridecanone (28.3%), and of CVOL α-pinene (34.8%), which were determined by gas chromatography. PPOL led juveniles to sedation at concentrations of 2 (703.8s), 10 (407.4s), 20 (225.5s), 50 (190.2s), and 80 (163.5s) mg L-1. CVOL also induced sedation at these concentrations, with times of 504.9, 294.2, 142.8, 113.8, and 135.6 s, respectively. Both EOs at 200 mg L-1 induced deep anesthesia within 507.5 (PPOL) and 1267.7 s (CVOL). Exposure to PPOL at 2, 20, and 80 mg L-1 for 48 h resulted in fish sedation, while CVOL lead fish to sedation at the two lowest concentrations and anesthesia at the highest one. Possible stress responses were evaluated during anesthetic recovery and long exposure, and PPOL did not cause behavioral or glucose levels changes in all evaluations. However, CVOL increased glucose levels at the highest concentrations studied (80 and 200 mg L-1), indicating that this oil may have caused stress in fish.

Abstract The sedative and anesthetic actions of several essential oils (EO) on fish have been demonstrated, stimulating the search for new options for natural anesthetics. This work evaluated the safety and sedative and anesthetic efficacy of EOs from three native Brazilian plants, Acmella oleracea (jambu), Aloysia hatschbachii and Cordia verbenacea (whale herb) in juvenile Rhamdia quelen (silver catfish). Anesthetic induction and recovery protocols (20 to 400 mg L-1) and long exposure (48 h) from 10 to 100 mg L-1 were tested. The EOs performed sedative and/or anesthetic activities: AOOi at a concentration of 20 mg L-1, AOOl at 50 and 100 mg L-1, AHOl, and CVOL (only sedation) 50 mg L⁻¹, as there were no important adverse effects and/or mortality. The results obtained indicate that Cordia verbenacea EO is the most promising as a sedative for juvenile silver catfish at a concentration of 50 mg L⁻¹.

Abstract This study aimed to evaluate the interactions between two low salinities (S = 4 and 8 g L−1) and two ionic balances (Na+/Mg+2/Ca+2/K+ ratio, IB1 = 27.6/3.6/1.05/1 and IB2 = 40.6/4.7/1.98/1) on zootechnical performance and histology of Litopenaeus vannamei postlarvae (0.0073 ± 0.0007 g) and juveniles (0.2163 ± 0.0080 g) reared in a biofloc system at stocking density of 1500 and 300 individuals m−3, respectively, over two culture phases: nursery (30 days) and grow‐out (60 days). High nitrification processes were detected at lower salinity treatments, with the lowest ammonia and nitrite concentration in those. There were no significant differences on shrimp survival between treatments; however, the best final weights were observed in the S8IB2 (0.24 ± 0.01 g) and S4IB1 (2.80 ± 0.12 g) treatments for the nursery and grow‐out stages, respectively. The most frequent histological lesions were the absence of lipid vacuoles in the hepatopancreas and abundant organic debris in the gill lamellae. The viability of rearing L. vannamei juveniles in biofloc at low salinity is still relatively weak because of their low growth and low efficiency to store fatty acids in the hepatopancreas; however, higher postlarvae productivity was found at 8 g L−1 and Na+/Mg+2/Ca+2/K+ ratio of 40.6/4.7/1.98/1 in the nursery stage.

Abstract The aim of this study is to analyze the waterborne metal levels in four lakes (one endorheic and three exorheic) of Harmony Point, Nelson Island, Antarctica. Water samples were analyzed by using a quadrupole type inductively coupled plasma mass spectrometer and an inductively coupled plasma optical emission spectrometer. The levels of As, Cu, Mn, Mo, and V were significantly lower and those of Cr, Mg, Na, and Sr were significantly higher in the endorheic lake than in the other lakes. Most water samples presented levels of Ag, Be, Cd, Pb, Se, Tl, and U below the limit of quantification, while for Ba, Co, and Ni around half of the samples were below this limit. The waterborne metal levels were not significantly different between the exorheic lakes. Waterborne metal levels in the freshwater lakes from Harmony Point did not show any clear relationship with their levels in the soil of the region or with bird guano, and overall, their levels indicate an environment without anthropogenic influence. Apparently, the Na levels are influenced by salt spray from the ocean, as they are related to the distance of the lakes from the ocean.

ABSTRACT: The current study tested the hypothesis that an increase in the ionic levels (Na+, Cl-, Ca2+, Mg2+, and K+) in the gastrointestinal tract (GIT), and hepatosomatic index (HSI), gonadosomatic index (GSI), and Fulton’s condition factor (FCF) are associated with the reproduction period of silver catfish (Rhamdia quelen). So, we verified ionic levels of the fluid phase of the different portions of the GIT content, HSI, GSI, and FCF of silver catfish in different seasons in a dam in southern Brazil. The highest Na+ and Cl- levels in GIT were found in spring. The stomach’s Na+ and Cl- values were lower than the other segments. The K+ and Ca2+ levels in GIT were higher in winter, except in the stomach, where the values increased in summer. The highest K+ levels were reported in the anterior intestine. The highest Ca2+ levels in winter and spring were reported in the anterior and mid-intestines. In summer, the Mg2+ levels in the intestine were lower. The values of GSI and FCF were higher in winter and spring, and those of HSI were higher in winter. In conclusion, ions in the GIT and HSI, GSI, and FCF were higher in the reproduction period (winter for spring).

Abstract This study evaluated the anesthetic activity of essential oil from Lippia alba (EOLA), linalool chemotype in a euryhaline fish (fat snook Centropomus parallelus). In the first experiment, fish were exposed to 30, 80, 130, 180, 200, and 230 µL EOLA L−1. The second experiment evaluated smaller and larger fish with 180 µL EOLA L-1. In the third experiment, ventilatory rates (VR) for up to 120 min in animals during exposure to 5 and 10 µL EOLA L−1 were evaluated. In the fourth experiment, fish anesthetized with 30 and 180 µL EOLA L-1 were assessed at 0, 30, and 60 min after anesthesia recovery to evaluate biochemical and antioxidant parameters. The best mild and deep anesthesia times were obtained with 30 and 180 µL EOLA L-1, and larger fish had the highest times. The VR increased in fish exposed to EOLA. Blood glucose and whole-body cortisol levels were higher in fish anesthetized with 180 µL EOLA L-1. Fish exposed to EOLA had higher liver glutathione S-transferase and superoxide dismutase activities without affecting catalase and lipid peroxidation levels. The 180 µL EOLA L-1 is recommended for fat snook anesthesia because it increases VR, blood glucose, and whole-body cortisol levels and prevents oxidative stress.

Abstract The growing increase in the fish farming sector has favored the establishment of bacterial outbreaks caused by Aeromonas hydrophila in several species. The hexane extract of Hesperozygis ringens (HEHR) (Lamiaceae) leaves increased the survival rate of silver catfish (Rhamdia quelen) experimentally infected by A. hydrophila. However, it is noteworthy that no reports have been found on the possible mechanisms of action of this extract in infected fish. This study aimed to evaluate the effect of the HEHR, administered through single immersion bath, on lipid peroxidation and antioxidant defenses in muscle and liver tissue of silver catfish challenged with A. hydrophila. The results showed that the oxidative status of silver catfish was altered, although oxidative stress was not triggered during the experiment. HEHR at 30 mg/L (HEHR30) was not characterized as a pro-oxidant agent in the presence of infection, unlike florfenicol and HEHR at 15 mg/L treatments in some cases. In short, HEHR30 provided an important increase in hepatic catalase activity, characterizing one of the possible mechanisms involved in the greater survival of fish experimentally infected by A. hydrophila. Additionally, HEHR30 did not induce lipid peroxidation, nor reduced antioxidant defenses of silver catfish infected or not by A. hydrophila.

Mugil liza juveniles (6.69 ± 0.06 g) were subjected to dietary citral (0-control, 0.5, 1.0 and 2.0 mL per kg feed) for 45 days, and its possible effects on zootechnical and metabolic parameters, digestive enzymes, innate immunity, oxidative status and liver damage were evaluated. At the end of the experiment, fish fed 2.0 mL citral per kg feed showed a greater weight gain and protein retention efficiency, as well as enhanced activities of pepsin (stomach) and amylase (intestine) compared with control fish. Citral supplementation decreased liver lipoperoxidation and increased the activities of glutathione peroxidase, glutathione-S-transferase and superoxide dismutase in the gills, liver and brain. The highest level of citral inclusion augmented non-protein thiol content in the brain and gills. Myeloperoxidase activity was lower in fish offered 1.0 and 2.0 mL citral per kg feed. Dietary citral did not influence the plasma levels of aspartate aminotransferase and alanine aminotransferase or the hepatic histology. As it improved growth, the activity of digestive enzymes and general health, dietary citral may be recommended for M. liza at 2.0 mL per kg feed.

ABSTRACT Lippia grata (formerly known as Lippia gracillis) is an aromatic plant native to Brazil, with leaves rich in essential oils that possess significant biological activities. We evaluated the effect of essential oil of L. grata (EOLG) as a dietary additive on the growth, somatic indices, and biochemical parameters of juveniles (5.25 ± 0.26 g) of tambatinga, a hybrid fish obtained by crossing tambaqui (Colossoma macropomum) with pirapitinga (Piaractus brachypomum) of great economic importance in north and northeastern Brazil. We evaluated four dietary treatments, consisting of EOLG supplemented at 0.0, 0.5, 1.0, and 2.0 mL kg-1, over 60 days. Carcass yield was significantly higher in fish fed all EOLG diets compared to those fed the control diet (0.0 mL kg-1). Animals that received the 0.5 mL kg-1 treatment gained significantly more weight and showed a higher specific growth rate than those treated with 1.0 and 2.0 ml kg-1 EOLG, although none differed significantly from the control. The feed conversion rate was significantly lower in the 0.5 than in the 1.0 mL kg-1 treatment. Compared with higher concentrations, the diet containing 0.5 mL kg-1 EOLG increased the use of muscle glycogen, glucose, and lactate to meet energy demands, avoiding the use of muscle protein. Our results suggest that dietary supplementation with EOLG significantly improves carcass yield in tambatinga juveniles but that concentrations above 0.5 mL kg-1 may compromise growth rates and carbohydrate metabolism in this fish.

This study aimed to identify the regions of the body surface of silver catfish (Rhamdia quelen) with a higher population of mucous cells in the skin. Additionally, the effects of stressful conditions, such as different stocking densities and food deprivation, on the proliferative response of mucous cells in the skin and gill epithelium and their impact on cutaneous mucous lysozyme activity were investigated. Silver catfish were divided into four experimental groups: high stocking density (32 kg/m3) and fed (HSD-F), high stocking density and fasted (HSD-FS), low stocking density (2.5 kg/m3) and fed (LSD-F), and low stocking density and fasted (LSD-FS). Fish in the fed groups received commercial feed twice a day, amounting to 1% of the tank biomass. After a 14-day experimental period, the fish were anesthetized and euthanized. Samples of cutaneous mucous and skin fragments from seven different points and the second left branchial arch were collected. Histological slides of the skin and gills were stained with PAS + Alcian Blue at pH 2.5, and the epidermal mucous lysozyme activity was assessed using the turbidimetric method. The ventral point in front of the ventral fin was found to be the optimal location for collecting cutaneous epithelia due to its higher density of mucous cells. The population of mucous cells in both the skin and gills varied based on the collection point and treatment applied. The highest lysozyme activity in the epidermal mucous was observed in fish from the HSD-F group. Overall, these findings suggest that stocking density and food deprivation create stressful conditions for silver catfish, which modulate their mucosal response to each situation.

R-(+)-limonene is a monoterpene from plants of the genus Citrus with diverse biological properties. This research evaluated the effects of dietary supplementation with R-(+)-limonene on growth, metabolic parameters in plasma and liver, and the antioxidant and stress responses in silver catfish, Rhamdia quelen, challenged or not with Aeromonas hydrophila. Fish were fed for 67 days with different doses of R-(+)-limonene in the diet (control 0.0, L0.5, L1.0, and L2.0 mL/kg of diet). On the 60th day, a challenge with A. hydrophila was performed. R-(+)-limonene in the diet potentiated the productive performance of the fish. The metabolic and antioxidant responses indicate that R-(+)-limonene did not harm the health of the animals and made them more resistant to the bacterial challenge. Histological findings showed the hepatoprotective effect of dietary R-(+)-limonene against A. hydrophila. Igf1 mRNA levels were upregulated in the liver of fish fed with an L2.0 diet but downregulated with bacterial challenge. The expression levels of crh mRNA were higher in the brains of fish fed with the L2.0 diet. However, the L2.0 diet downregulated crh and hspa12a mRNA expression in the brains of infected fish. In conclusion, the results indicated that R-(+)-limonene can be considered a good dietary supplement for silver catfish.

The present study evaluated different levels of the essential oil of Nectandra grandiflora (EONG) in the diet of juvenile Colossoma macropomum. The juveniles (0.75 ± 0.05 g) were fed four experimental diets with differing levels of EONG (0, 0.5, 0.75, and 1.50 mL/kg). After 20 days, the weight, daily weight gain, specific growth rate, and daily feed consumption per fish were highest for the fish fed 0.5 mL EONG/kg, while after 30 days, only the weight and daily feed consumption per fish for that diet remained highest. The viscerosomatic index was highest for the fish fed 0.5 mL EONG/kg while the hepatosomatic index was higher for the groups that received dietary EONG for 30 days. The juveniles fed 1.5 mL EONG/kg showed a proliferative response of the mucous cells in the gills, but the proliferation of these lysozyme- and immunoglobulin-secreting mucosal cells was higher for the skin of the fish of all EONG levels, compared to that of the control fish. The fish fed 0.5 mL EONG/kg had the greatest height and width of intestinal villi. The two highest levels of dietary EONG supplementation (0.75 and 1.50 mL/kg) reduced the hepatocyte dimensions but did not modify the centrolobular vein area. Dietary supplementation with 0.5 mL EONG/kg improved the growth, gut health, and immune response of juvenile C. macropomum.

ABSTRACT: This study evaluated the effectiveness of the essential oil of Hesperozygisringens (EOHR) for anesthesia of Colossomamacropomum by documenting hematological and blood biochemical responses after biometric handling. In Experiment 1, juveniles (14.12 ± 3.53 g) were exposed to different concentrations of EOHR: 0 (control), 75, 150, 300 and 450 µL L-1 (n=10 fish for each concentration), to determine times for induction and recovery from anesthesia, as well as its effects on ventilatory frequency (VF). Based on these results, Experiment 2 evaluated the effects of 0 (control), 75 (with induction and recovery times outside that recommended for fish anesthesia) and 150 µL L-1 EOHR (within recommended times) on hematological and biochemical variables of juveniles (20.52 ± 3.47 g) after anesthesia and after 24 h of recovery (n = 6 fish for each concentration and collection time). Survival was 100%. Induction time showed a quadratic effect of EOHR concentration. Recovery time did not differ among EOHR concentrations. Concentrations between 150 and 450 µL L-1 EOHR caused rapid induction (< 3 min) and recovery (< 5 min). EOHR concentration affected VF. The concentration of 150 µL L-1 EOHR had little influence on hematological and biochemical parameters of C. macropomum of 20 g.

This study aimed to evaluate the supplementation of peppermint essential oil (Mentha piperita; PEO) in diets for juvenile Nile tilapia (Oreochromis niloticus). A feeding experiment with diets containing graded levels (0.0, 0.2, 0.4, 0.6, 0.8, and 1.0 g kg−1) of PEO was carried out with fish of 0.58 ± 0.08 g. The inclusion of graded levels of PEO in the diet improved the weight gain (0.52 g kg−1), feed intake (0.51 g kg−1), and feed conversion (0.51 g kg−1) of juvenile Nile tilapia in a quadratic pattern. Increasing levels of PEO also led to a linear decrease in body lipid content. The gut activity of the digestive enzymes amylase (0.54 g kg−1) and protease (0.39 g kg−1) increased quadratically, whereas lipase activity increased linearly. PEO increased the activity of the antioxidant enzymes catalase (CAT; 0.52 g kg−1) and superoxide dismutase (SOD; 0.58 g kg−1) while reducing the production of malonaldehyde (MDA; 0.40 g kg−1) and nitric oxide (NO; 0.63 g kg−1) in the liver. The results of this study provide evidence of the beneficial effects of PEO on the growth and health of Nile tilapia and recommend a dose of 0.6 g kg−1 as the optimal level of supplementation.

Amphipods are frequently used as bioindicators of water quality in experimental or behavior trials. Thus, it is a group considered suitable for use as a model organism in tests with essential oils (EOs). This study evaluated the time required for anesthesia induction and recovery of the amphipod crustacean H. bonariensis exposed to the essential oils of Aloysia triphylla (EOAT) and Lippia alba (EOLA), and their major compounds citral and linalool, respectively. In addition, we evaluated the locomotor activity of amphipods using ANY-maze® software. Mortalities were observed at concentrations of 100 and 200 µL/L of citral (50.0 ± 0.39%) and 750 µL/L of EOLA (66.7 ± 0.33%). Except for linalool, increased concentrations of the compounds of the essential oils decreased the time for sedation and anesthesia induction. There were differences for the induction of anesthesia (p < 0.05) and recovery (p < 0.05) between EOLA and linalool treatments, but not between that for EOAT and citral. Reduced locomotor activity and longer time and episodes of freezing were observed in animals exposed to EOAT. The EOs and their major compounds induced anesthesia and affected the locomotor activity of H. bonariensis, Therefore, EOAT and linalool are recommended for anesthesia of this species. EOAT can also be utilized in long-term exposure.

Creatine kinase (CK) is an enzyme that produces and uses phosphocreatine to transfer energy to maintain tissue and cellular energy homeostasis, being considered the main controller of cellular energy homeostasis. Its activity in plasma/serum has been commonly used to evaluate tissue damage, since CK is released into the bloodstream during damage. This review summarizes the current knowledge regarding the use of CK activity in fish, focusing on its potential as an indicator of the impairment of energetic homeostasis and tissue damage during stressful situations, such as exposure to contaminants (metals, pesticides, microplastic), hypoxia, thermal stress, and diseases (fungal, parasitic, and bacterial). Based on the data, we can conclude that tissue CK activity can be used as a suitable indicator of the impairment of energetic homeostasis in fish exposed to different aquaculture challenge conditions, while serum/plasma CK activity can be used as the first evidence of possible tissue damage, due to its release into the bloodstream.

Abstract The objective of this study was to evaluate the possible relaxing effect of essential oils (EOs) (Aloysia triphylla and Lippia alba) and phytochemicals (citral and linalool) in the gastropod Pomacea canaliculata. Animals were exposed to compounds at the concentrations range of 25-750 µL L–1. Magnesium chloride (MgCl2, 10-50 g L–1) and control group (ethanol 6.75 mL L–1, highest concentration used for treatment dilution) were also tested. The EOs, citral and MgCl2 had no relaxing effect at the concentrations range tested, and citral caused aversive behavior (closure of the operculum) from 90 μL L–1. Exposure to linalool at 25, 50, 100, 200 and 400 µL L-1 relaxed 28, 76, 88, 96 and 100% of the animals, respectively. The concentrations of 25, 50 and 400 µL L-1 differed statistically from each other, while 100 and 200 µL L-1 were equal to 50 and 400 µL L-1. All animals recovered up to 40 min, except at of 400 µL L-1. Linalool is effective for relaxing P. canaliculata and can be useful in management techniques that require relaxation. However, further studies are needed to certify whether linalool is appropriate for maintaining animal welfare in invasive procedures that require total insensitivity.

ABSTRACT: The use of anesthetics in aquaculture ensures better animal welfare and survival during transport and the production cycle. The present study evaluated the anesthetic efficacy of essential oils (EOs) of Lippia alba (EOLA) and Ocimum gratissimum (EOOG) for pink shrimp Farfantepenaeus paulensis, and EOs of Origanum majorana (EOO) and Cymbopogon citratus (EOC) for white shrimp Litopenaeus vannamei. shrimp were exposed to (i) 500, 750 or 1000 µL L-1 of EOLA or (ii) 50, 100, 150 or 250 µL L-1 of EOOG, and L. vannamei were exposed to (iii) 150, 300 or 500 µL L-1 of EOC or (iv) 400 or 800 µL L-1 of EOO. The induction times were concentration dependent, with a decrease in induction time with the increase of the EOs concentration, but for EOLA, this pattern was observed only for anesthesia. Induction times for sedation and anesthesia were significantly shorter for shrimp exposed to EOC and EOO. The highest concentration of EOOG (250 µL L-1) resulted in 30% mortality. The recovery time was significantly longer for shrimp exposed to 800 µL L-1 of EOO compared to the other EOs. Overall, the action of EOs significantly differed between the two shrimp species. In conclusion, the tested EOs effectively anesthetized F. paulensis and L. vannamei.

Exposure to acidic and alkaline pHs results in an ionic imbalance. Cellular responses involved in osmoregulation in silver catfish exposed to different pHs (5.5, 7.5, and 9.0) for 24 h were evaluated. The gills and kidney were collected to measure Na+/K+-ATPase (NKA) and H+-ATPase (V-ATPase) activities and to evaluate the expression of ion transporter-related genes: NKA (atp1a1), H+-ATPases (atp6v0a1b, atp6v0a2a, atp6v0a2b), Na+/H+ antiporter (slc9a3), K+/Cl− symporters (slc12a4, slc12a6, slc12a7a, slc12a7b), Na+/K+/2Cl− symporter (slc12a2), and ammonium transporter Rh type b (rhbg). The gills presented greater responses to pH changes than the kidney. The pH alterations changed the atp1a1 gene expression and NKA activity, whereas the H+-ATPase activity increased in the gills in alkaline water, probably to maintain ionic balance. The slc9a3 and slc12a2 genes play more prominent roles in the ion uptake at acidic pH than H+-ATPase. The slc12a7a was the only isoform of this transporter affected by pH. The rhbg is apparently related to ammonia excretion through the gills and kidney (minor scale). Exposure to alkaline pH seems to be battled by impairment of NKA and H+-ATPase activities in the gills, whereas the expression of some ion transporters in silver catfish changes during both acidic and alkaline pHs.

Cinnamon essential oils (EOs) are widely known for their pharmaceutical properties; however, studies investigating the use of these EOs in aquaculture are scarce. The aims of this study were to evaluate the anesthetic effect of bathing silver catfish (Rhamdia quelen) with Cinnamomum cassia EO (CCEO) and its nanoemulsion (NCCEO); the growth-promoting and antioxidant effects of dietary supplementation with CCEO in silver catfish; and the in vitro antibacterial effect of CCEO, NCCEO, and Cinnamomum zeylanicum EO (CZEO) against bacteria isolated from diseased silver catfish. The two cinnamon EOs showed promising antibacterial activity, which was potentiated by the nanoemulsion. CCEO showed satisfactory anesthetic activity in silver catfish, and its nanoemulsion intensified the sedative activity. Supplementation of 1.0 mL CCEO per kg of diet for 60 days increased weight, length, and weight gain when compared to the control group, evidencing the growth-promoting activity of this EO. Dietary supplementation of CCEO for 30 and 60 days also showed an antioxidant effect, as it decreased levels of thiobarbituric acid reactive species and increased the superoxide dismutase activity in the liver of silver catfish. Therefore, cinnamon EOs have a promising use in aquaculture.

Advances in fisheries and aquaculture science often follow the introduction of new tools or analytic methods [...]

ABSTRACT: Essential oils of plants whose main compound is citral showed beneficial effects when added to fish feed. The objective of the present study was to evaluate the dietary effect of the addition of citral on zootechnical parameters and digestive enzyme activities of Centropomus undecimalis. Juveniles were fed for 45 days with diets containing different amounts of citral (0.0 - control, 0.5, 1.0, and 2.0 mL per kg of diet). The water quality parameters were kept stable during the experiment and no mortality was observed. At the end of the experimental period, the treatment 0.5 mL citral per kg of diet had the lowest weight gain and specific growth rate, and the highest feed conversion, while the same parameters did not differ between the other treatments. Pepsin activity was higher in the stomach of fish fed with 0.5 mL citral per kg of diet and amylase activity was higher in the intestine of fish fed with 0.5 and 2.0 mL citral per kg of diet compared to the control group. Intestinal lipase activity was higher in all groups that were fed with citral compared to the control group. Chymotrypsin and trypsin activities showed no difference between groups. Consequently, dietary addition of citral at any of the levels tested is not recommended for common snook.

ABSTRACT: The objective of this study was to evaluate the efficacy of the citral chemotype of Lippia alba essential oil (EOLA) as an additive in a simulated transport with different loading densities of tambaqui (Colossoma macropomum) juveniles. Simulated transport of tambaqui juveniles at three loading densities (30, 60 and 90 fish/L) was carried out either with only water (WC) or 20 μL.L-1 EOLA. Dissolved oxygen levels, pH, conductivity and temperature of the water were measured at the beginning and end of the simulated transport. Treatment with EOLA did not change survival or dissolved oxygen levels at the end of simulation. The pH of the water at the end of simulation with EOLA was lower than in that of WC but did not change with loading density. Water conductivity increased with loading density; however, it was lower in the groups with EOLA. The addition of EOLA can be useful in long-term transportation at high loading densities, but further studies related to tambaqui physiology are required.

ABSTRACT The aim of this study was to evaluate the efficacy and safety of several anesthetics in the brachyuran crab Neohelice granulata, an emergent experimental model. The essential oils (EOs) of Lippia alba, Aloysia tryphilla, and Melaleuca alternifolia (tea tree oil; TTO), the isolated compounds eugenol, menthol, terpinen-4-ol, and the nanoencapsulated form of TTO, were administered in one or more of the following ways: added to the water (immersion), through an arthrodial membrane (injected), or by oral gavage. Unexpectedly, most EOs did not produce an anesthetic effect after immersion. Only TTO and eugenol induced anesthesia by immersion, with very long induction and recovery times compared to anesthesia of other crustaceans. However, a good anesthetic effect was observed with the injection of terpinen-4-ol and nanoencapsulated TTO in N. granulata; both demonstrated ideal induction and recovery times. These substances appear to be promising anesthetic alternatives for crustaceans.

Lactococcosis, particularly that caused by Lactococcus garvieae, is a major re-emerging bacterial disease seriously affecting the sustainability of aquaculture industry. Medicinal herbs and plants do not have very much in vitro antagonism and in vivo disease resistance towards lactococcosis agents in aquaculture. Most in vitro studies with herbal extractives were performed against L. garvieae with no strong antibacterial activity, but essential oils, especially those that contain thymol or carvacrol, are more effective. The differences exhibited by the bacteriostatic and bactericidal functions for a specific extractive in different studies could be due to different bacterial strains or parts of chemotypes of the same plant. Despite essential oils being shown to have the best anti-L. garvieae activity in in vitro assays, the in vivo bioassays required further study. The extracts tested under in vivo conditions presented moderate efficacy, causing a decrease in mortality in infected animals, probably because they improved immune parameters before challenging tests. This review addressed the efficacy of medicinal herbs to lactococcosis and discussed the presented gaps.

“A new journey for Fishes” [...]

Brazilian fish farming goes together with the emergence of numerous bacterial diseases, with Aeromonas hydrophila being the main bacterial pathogen. As a consequence, antimicrobials are excessively used. Considering that antimicrobials are relatively stable and nonbiodegradable, medicinal plants and their phytochemicals have been used as alternative treatments of bacteriosis in fish farming. Limonene is a monoterpene available in two enantiomers: R-(+)-limonene and S-(-)-limonene. This study analyzed the antibacterial activity of the phytochemicals S-(-)-limonene and R-(+)-limonene against some bacteria isolated from silver catfish (Rhamdia quelen). Furthermore, by means of spectrophotometry and atomic force microscopy, we also investigated the combination therapy of phytochemicals with antimicrobials and their activity in terms of inhibiting biofilm formation. Six clinical isolates and a standard strain were selected for antimicrobial activity testing. Biofilm formation was tested in 96-well plates and nylon cubes. The most sensitive of the strains tested was the A. hydrophila strain (MF 372510). S-(-)-limonene and R-(+)-limonene had high minimum inhibitory concentrations; however, they strongly inhibited A. hydrophila biofilm formation. R-(+)-limonene and S-(-)-limonene had an additive effect when combined with florfenicol and an antagonistic effect with oxytetracycline. In general, the phytochemicals tested showed strong antibiofilm activity against A. hydrophila, and when in combination therapy with florfenicol, they showed an additive effect against the treatment of A. hydrophila.

Maclura tinctoria is a tree species native from Brazil and rich in phenolic compounds. Since plant antibacterial activity is highly associated with phenolic compound concentration, we aim to evaluate the in vitro antimicrobial activity of different extracts against fish pathogenic bacteria. In addition, some phenolic compounds have central depressant effects and can be useful in aquaculture due to possible sedative and/or anesthetic effects. Four M. tinctoria extracts were extracted separately with ethanol; leaves (LE), bark (BE), heartwood (HE), and the sapwood (SE). In vitro antimicrobial activity was tested against Aeromonas strains at concentrations of 6400 to 3.125 μg/mL. The sedative effect was evaluated for 24 h with 30 and 100 mg/L concentrations. Chemical composition was analyzed by HPLC-DAD-MS. The HE extract had the best MIC (400 µg/mL) and MBC (800 µg/mL) compared to the LE, BE, and SE extracts. LE extract induced deep sedation and the BE, SE, and HE extracts induced light sedation. Additionally, BE, SE, and HE induced a normal behavior without side effects. Polyphenolic compounds with antimicrobial activity and sedative effects were identified mainly in HE. Thus, HE extract is safe and can be used as a sedative for silver catfish.

ABSTRACT This study aimed to assess the effects of low and high water hardness in interaction with different water pH in pacu (Piaractus mesopotamicus). Pacu juveniles were subjected to low (50 mg CaCO3 L-1 - LWH) or high water hardness (120 mg CaCO3 L-1 - HWH) at water pH of 5.5 (acidic), 7.5 (circumneutral) or 9.0 (alkaline) for 15 days. Gills and kidneys were collected (days 1, 5 and 15). Gill Na+/K+-ATPase (NKA) and vacuolar-type H+-ATPase (V-ATPase) activities were higher in alkaline pH with HWH on day 1. Gill and kidney NKA and V-ATPase activities were higher in acidic pH with LWH on day 15. Gill NKA activity of pacus under alkaline pH with LWH was higher than those exposed to HWH. Reduced antioxidant capacity in the gills and kidney and enhanced thiobarbituric acid reactive substances (TBARS) levels were demonstrated in fish exposed to acidic or alkaline pH, mainly with LWH. HWH increased glutathione-S-transferase (GST) activity and reduced TBARS levels in the gills and kidney. On day 15, GST activity was increased at acidic pH with LWH. In conclusion, circumneutral pH presents less oxidative stress and fewer variations in ATPases and HWH reduced deleterious effects in fish exposed to acidic or alkaline pH.

ABSTRACT: The growth of cachama hybrids, cachamoto (Colossoma macropomum ♀ x Piaractus brachypomus ♂) and cachamay (P. brachypomus ♀ x C. macropomum ♂) was compared for 154 days to determine which showed the better productivity. The experiment was carried out in three earth ponds, each divided transversely with a plastic net, yielding six experimental units. Juveniles of cachamoto of 12.95±2.43g and 6.75±0.42cm and cachamay of 16.65±3.64g and 7.41±0.62cm were distributed in each sub-tank of 240m2 at a density of 0.5 fish m-2. Fish were fed twice daily except on days when their weight and length, and the physical and chemical parameters of the water were measured. The following productive variables were evaluated: growth in weight and length, daily weight gain, daily feed intake, apparent feed conversion, specific growth rate, condition factor, productivity and the cost/benefit relationship. None of the analyzed variables presented statistical difference between treatments. Physical and chemical parameters of the water remained within the recommended values for cachama, and survival was 100%. Based on the results obtained, it can be concluded that cachama reciprocal hybrids present a similar productive behavior, so that it is indifferent which one is chosen to be raised under the conditions evaluated.

Citral is a monoterpenoid found in the essential oils of several plant species, recognized as a safe diet additive and extensively used in the food and cosmetic industries. This study evaluated the effects of dietary supplementation with citral (0.25 g/kg), loaded in nanosystems or unloaded on the gastrointestinal tract (GIT) pH, hepatosomatic and intraperitoneal fat index, innate immune responses and gut microbiota (GM) of silver catfish (Rhamdia quelen). The treatments were: control diet (CO), diet containing unloaded citral (UC), diet containing citral-loaded nanoemulsions (NE), and diet containing citral-loaded alginate nanoparticles (NP). After 21 days fish fed all diets containing citral showed an increased liver lysozyme activity, whereas fish fed UC showed reduced serum hemolytic activity and intestinal myeloperoxidase activity. Serum lysozyme activity was lower in fish fed diet containing NP, whereas fish fed NE showed a reduced blood macrophage phagocytic index. Citral reduced stomach pH (all delivery forms) and increased intestinal pH when loaded in NE. Both NE and NP reduced the total bacterial population in the fish intestine, whereas Aeromonas spp. in intestine were higher in fish fed diet containing NP. Citral modulated the silver catfish immune responses, GIT pH and GM in a delivery mode-dependent manner.

In fish, stressful events initiate a hormone cascade along the hypothalamus-pituitary-interrenal and hypothalamus-sympathetic-chromaffin (HSC) axis to evoke several physiological reactions in order to orchestrate and maintain homeostasis. Several biotic and abiotic factors, as well as aquaculture procedures (handling, transport, or stocking density), activated stress system inducing negative effects on different physiological processes in fish (growth, reproduction, and immunity). In order to reduce these consequences, the use of essential oils (EOs) derived from plants has been the focus of aquaculture studies due to their diverse properties (e.g., anesthetic, antioxidant, and antimicrobial), which have been shown to reduce biochemical and endocrine alterations and, consequently, to improve the welfare status. Recently, several studies have shown that biogenic compounds isolated from different EOs present excellent biological activities, as well as the nanoencapsulated form of these EOs may potentiate their effects. Overall, EOs presented less side effects than synthetic compounds, but their stress-reducing efficacy is related to their chemical composition, concentration or chemotype used. In addition, their species-specific actions must be clearly established since they can act as stressors by themselves if their concentrations and chemotypes used are not suitable. For this reason, it is necessary to assess the effect of these natural compound mixtures in different fish species, from marine to freshwater, in order to find the ideal concentration range and the way for their administration to obtain the desired biological activity, without any undesired side effects. In this review, the main findings regarding the use of different EOs as stress reducers will be presented to highlight the most important issues related to their use to improve fish welfare in aquaculture.

ABSTRACT: The use of natural products, such as essential oils (EOs), is a potential novel approach to treat fish bacterial infections with a lower risk of developing resistance. There has been a number of studies reporting the activity of EOs as those obtained from the species Achyrocline satureioides, Aniba parviflora, Aniba rosaeodora, Anthemis nobilis, Conobea scoparioides, Cupressus sempervirens, Illicium verum, Lippia origanoides, and Melaleuca alternifolia against bacteria. However, there are few studies investigating the effect of these EOs against fish bacteria. Therefore, the aim of this study was to evaluate the in vitro antibacterial activity of EOs against the following fish bacteria, Aeromonas hydrophila, Citrobacter freundii, and Raoultella ornithinolytica. Additionally, the in vivo antibacterial activity of the EO L. origanoides was evaluated against experimentally induced A. hydrophila infection of silver catfish (Rhamdia quelen). The EO of L. origanoides was chosen as it showed the highest in vitro antibacterial activity, with minimum inhibitory concentrations ranging from 0.2 to 0.8 mg mL-1. This EO also presented a therapeutic success of 58.33%, on a 30 day A. hydrophila infection. Therefore, we suggested that the EO of L. origanoides may be a viable alternative as a treatment for A. hydrophila infection.

ABSTRACT: Anesthetics are effective to reduce or minimize stress in handling operations related to fish cultures. Aiming to avoid residual effects of synthetic drugs, several natural agents, such as essential oils (EOs), have been tested. The aim of this study was to determine the optimal concentration of the EO of Ocimum micranthum for induction of anesthesia in juveniles of silver catfish (Rhamdia quelen) and grass carp (Ctenopharyngodon idella). Results demonstrated that the most suitable concentration to sedate silver catfish and carp is 25μL/L, while to anesthetize 200μL/L is more suitable for silver catfish and 100μL/L for grass carp, because these concentrations induce deep anesthesia in less than three min and recovery in less than five min. Analysis of the EO constituents showed methyl chavicol, a compound with carcinogenic potential, as the major component (58.2%). Therefore, the EO of O. micranthum is not recommended for fish anesthesia if the fish are to be used as food.

ABSTRACT: Humic substances are organic compounds that possess high molecular weight and are formed by the decomposition of plant materials. Humic substances comprise humic acids, fulvic acids and humin. Humic acids (HA) have shown to be able to provide some protection to biological membranes of fish in water with low pH, replacing the protective action of Ca2+. Thus, this studyaimed to observe growth of silver catfish ( Rhamdia quelen ) juveniles exposed to pHs 5.5 and 6.5 and to different levels of HA: 0, 10, 25 and 50mg L-1. Results showed that, irrespective of pH, HA was highly detrimental to silver catfish growth, since the higher the concentration of HA, the lower the weight gain and feed intake, resulting in lower biomass and lower specific growth rate of juveniles. Hence, HA is extremely damaging to the performance of silver catfish juveniles in the tested concentrations.

ABSTRACT: This research aimed to evaluate whether the essential oil of Aloysia triphylla (EOAT) used in vivo as a sedative in the water for transporting fish could increase the oxidative stability of silver catfish (Rhamdia quelen) fillets during frozen storage. The chemical composition of EOAT and of fillets from fish exposed to EOAT (0, 30 or 40µL L-1) were assessed. The pH and lipid oxidation parameters (conjugated dienes, CD; thiobarbituric acid-reactive-substances, TBARS) were evaluated in the fillets throughout the storage period (-18±2oC/17 months). The main compounds found in EOAT were α- and β-citral. Treatment with EOAT did not modify the proximate composition of the fillets, but 40µL L-1 EOAT reduced pH levels when compared to the control fillets (P

ABSTRACT This study investigated the efficacy of essential oils of Lippia alba (EOLA) citral chemotype and Lippia origanoides (EOLO) and their effects on ventilatory frequency (VF) of silver catfish, Rhamdia quelen. Fish were exposed to 50-300 μL L−1 of EOLA and 25-300 μL L−1 of EOLO to determine induction times to sedation, anesthesia, and recovery. Moreover, VF was determined in fish exposed to 5 or 10 μL L−1 of EOLA and of EOLO for 8 h. The increasing concentration of essential oils proportionally decreased sedation and anesthesia induction times. The highest EOLA concentration increased VF of fish from the control group at 1 h of exposure, but VF decreased at both EOLO concentrations after 2 h. The EOLA citral chemotype and EOLO were effective sedatives and anesthetics for silver catfish. However, EOLO was the most suitable sedative for additional studies regarding fish transport as it reduced VF and did not induce VF increase in silver catfish. The EOLA citral chemotype and EOLO are effective sedatives and anesthetics for silver catfish. Moreover, the EOLO is recommended for transport of silver catfish, because it maintains the ventilatory frequency constant, avoiding a possible metabolic stress.

ABSTRACT This study investigated the anesthetic effect of the essential oils (EOs) from the peel of Citrus x aurantium (EOCA) and Citrus x latifolia (EOCL) on silver catfish Rhamdia quelen. Fish were exposed to different concentrations of EOCA and EOCL to determine time of anesthesia induction and recovery. Induction of anesthesia was observed in all fish exposed to 400, 600 or 800 μL L−1 EOCA and 300, 400 or 500 μL L−1 EOCL. Another group of fish were exposed for 8 h to 50, 100, or 200 μL L−1 of either EOs. Overall, fish exposed to ethanol and both EOs presented higher ventilatory frequencies (VF) than the control group throughout the 8 h of exposure. Net ion (Na+, K+ and Cl−) effluxes and ammonia excretion were significantly lower in fish exposed to 50, 100 or 200 μL L−1 of either EOs compared to control fish. Mortality was 37% in fish exposed to 200 μL L−1 of either EOs after 8 h. These findings suggest that EOCA and EOCL are useful anesthetics and sedatives for Rhamdia quelen, but their usefulness as alternatives to reduce stress in fish transportation at the lower concentrations tested (50-100 µL L−1) deserves further study.

ABSTRACT The aim of this study was to determine the composition of gastrointestinal content and protease and lipase activities in summer and winter as well as to evaluate the relationship between digestive enzyme activity and centesimal composition of gastrointestinal content and feeding habits of two omnivorous species, Rhamdia quelen (Quoy & Gaimard, 1824) and Pimelodus maculatus (LaCèpede, 1803), and of two detritivorous species, Loricariichthys anus (Valenciennes, 1835) and Hypostomus commersoni (Valenciennes, 1836). The activities of pepsin, trypsin, chymotrypsin, and lipase, and the levels of proteins and lipids in the gastrointestinal tract, were evaluated. The enzyme activities were not related to the centesimal composition of gastrointestinal content or feeding habits. This finding could be associated with the variations of nutrient availability over time in the environment, as was observed in the centesimal composition of food ingested by the fish in summer and winter. The analyzed enzymes exhibited a constitutive character in these species; that is, the digestive enzymes are always available in the gastrointestinal tract to digest any food that the fish may find as an adaptation to better utilize the nutrients available in the environment in winter and summer.

ABSTRACT The aim of this study was to determine the anesthetic efficacy of the essential oils (EOs) of Aniba rosaeodora (EOAR) and Aniba parviflora (EOAP) and one of their main compounds, linalool, in two forms: synthetic and extracted from EOAR (linalool-AR) in tambaqui (Colossoma macropomum). In the first experiment, the anesthetic induction and recovery of juveniles exposed to 25- 200 µL L-1 of EOAR or 50- 300 µL L-1 of EOAP or synthetic linalool or linalool-AR was evaluated. The second experiment observed the behavioral effects of long-term exposure (12h) of these EOs and linalools (5 and 10 µL L-1). Fish exposed to 50-200 µL L-1 of EOAR and 100-300 µL L-1 of EOAP and both linalools reached deep anesthesia between 1-10 min. Induction time for all anesthesia stages decreased with the increasing concentration of the anesthetics. Linalool-AR showed lengthier time for anesthesia induction in some stages and for recovery at 100 and 200 µL L-1 in comparison to synthetic linalool. Normal equilibrium and swimming behavior was observed in fish exposed to the EOs and linalools throughout the 12 h of exposure. In conclusion, both EOs and linalools can be used as anesthetics and sedatives in tambaqui.

The objective of this study was to evaluate the tolerance of piavas (Leporinus obtusidens) to ammonia (NH3) by measuring its effects on plasma ion levels and tissue metabolic parameters. Piava juveniles (25-30 g) were exposed to five concentrations of NH3 (mg L-1): 0.003 (control), 0.1, 0.4, 0.7, and 1.4; after 96 hours, plasma levels of Na+, K+, Cl- and NH4+ ions and metabolic and enzyme activity in tissues (liver, kidneys, gills and muscle) were measured. The lethal concentration (LC50; 96h) of NH3 was 0.27 mg L-1. As NH3 increased, Na+ and NH4+ in the plasma increased and K+ decreased. In addition, Na+/K+- ATPase activity concomitantly increased in the gills and decreased in the kidneys. Glucose, glycogen, and protein levels decreased, while lactate and ammonia increased in the tissues of piava juveniles that were treated with higher concentrations of ammonia. The observed lethal toxicity could be due to a gradual depletion of plasma ion levels and a reduction of metabolic and Na+/K+-ATPase activity in tissues. Both can lead to dysfunction in ionoregulatory and physiological systems. This finding has implications for the management of fish culture of piavas.

The objective of this study was to verify net ion fluxes and ammonia excretion in silver catfish transported in plastic bags at three different loading densities: 221, 286 and 365g L-1 for 5h. A water sample was collected at the beginning and at the end of the transport for analysis of water parameters. There was a significant positive relationship between net ion effluxes and negative relationship between ammonia excretion and loading density, demonstrated by the following equations: Na+: y-24.5-0.27x, r2=0.99, Cl-: y=40.2-0.61x, r2=0.98, K+: y=8.0-27.6x, r2=0.94; ammonia excretion: y=-11.43+0.017x, r2=0.95, where y: net ion flux (mmol kg-1 h-1) or ammonia excretion (mg kg-1h-1) and x: loading density (g). Therefore, the increase of loading density increases net ion loss, but reduces ammonia excretion during the transport of silver catfish, indicating the possibility of ammonia accumulation

This work aimed to determine the efficacy of the essential oil of A. gratissima as anesthetic for silver catfish, and to perform the bio-guided fractionation of essential oil aiming to isolate compounds responsible for the noted effects. Fish were submitted to anesthesia bath with essential oil, its fractions and isolated compounds to determine time of anesthetic induction and recovery. Eugenol (50 mg L-1) was used as positive control. Essential oil of A. gratissima was effective as an anesthetic at concentrations of 300 to 900 mg L-1. Fish presented involuntary muscle contractions during induction and recovery. The bio-guided fractionation of essential oil furnishedE-(-)-pinocamphone, (-)-caryophyllene oxide, (-)-guaiol and (+)-spathulenol. E-(-)-pinocamphone caused the same side effects observed for essential oil. (-)-Caryophyllene oxide, (-)-guaiol and (+)-spathulenol showed only sedative effects at proportional concentrations to those of the constituents in essential oil. (+)-Spathulenol (51.2 mg L-1) promoted deep anesthesia without side effects. A higher concentration of (+)-spathulenol, and lower or absent amounts ofE-(-)-pinocamphone could contribute to increase the activity and safety of the essential oil of A. gratissima. (+)-Spathulenol showed potent sedative and anesthetic activities in silver catfish, and could be considered as a viable compound for the development of a new anesthetic.