Protective effects of diet containing rutin against trichlorfon-induced muscle bioenergetics disruption and impairment on fatty acid profile of silver catfish Rhamdia quelen.

Trichlorfon is an organophosphate insecticide that is widely used on fish farms to control parasitic infections. It has been detected in freshwater ecosystems as well as in fishery products. There is a growing body of evidence to suggest that certain feed additives may reduce or prevent pesticide-induced toxicity in fish. The aim of the present study was to determine whether acute exposure to trichlorfon would alter bioenergetic homeostasis and alter fatty acid profiles in muscles of silver catfish (Rhamdia quelen). We also sought to determine whether rutin prevents or reduces these effects. Cytosolic and mitochondrial creatine kinase (CK) and activities of complexes II-III and IV in muscle were significantly inhibited by exposure to 11 mg/L trichlorfon for 48 h compared to effects in the unexposed group. Total content of polyunsaturated fatty acids (omega-3 and omega-6) were significantly lower in muscle of silver catfish exposed to 11 mg/L trichlorfon for 48 h than in the unexposed group. Addition of 3 mg rutin/kg feed increased CK activity and prevented inhibition of complex IV activity, as well as preventing all alterations of muscle fatty acid profiles elicited by exposure to trichlorfon. No significant differences were observed between groups with respect to muscle adenylate kinase or pyruvate kinase activities, as well as total content of saturated and monounsaturated fatty acids. Our findings suggest that exposure (48 h) to 11 mg trichlorfon/L water inhibits cytosolic and mitochondrial CK activity in muscle. Trichlorfon also affects activities of complexes II-III and IV in respiratory chain, with important consequences for adenosine triphosphate production. The pesticide alters fatty acid profiles in the fish and endangers human consumers of the product. The most important finding of the present study is that inclusion of rutin improves bioenergetic homeostasis and muscle fatty acid profiles, suggesting that it reduces trichlorfon-induced muscle damage.

Diphenyl diselenide dietary supplementation protects against fumonisin B1-induced oxidative stress in brains of the silver catfish Rhamdia quelen.

The trend toward using plant-based ingredients in aquafeeds has raised important concerns for aquaculture owing to the negative impacts of mycotoxins on fish health; with emphasis for contamination by fumonisin B1 (FB1). The brain is an important target of FB1; however, study of the pathways linked to brain damage is limited to an analysis of histopathological alterations. Reports have demonstrated the protective effects of dietary supplementation with diphenyl diselenide (Ph2Se2) in the brains of fish subjected to several environmental insults; nevertheless, its neuroprotective effects in fish fed with diets contaminated with FB1 remain unknown. Therefore, the aim of this study was to evaluate whether oxidative damage may be a pathway associated with FB1-induced neurotoxicity, as well as to evaluate whether dietary supplementation with Ph2Se2 prevents or reduces FB1-mediated brain oxidative damage in silver catfish. Brain reactive oxygen species (ROS), lipid peroxidation (LOOH) and protein carbonylation increased on day 30 post-feeding in animals that received FB1-contaminated diets compared to the control group, while brain antioxidant capacity against peroxyl radicals (ACAP) levels and catalase (CAT), glutathione peroxidase (GPx) and glutathione S-transferase (GST) activities were lower. Diphenyl diselenide dietary supplementation avoid increases in brain ROS levels, as well minimizing the augmentation of LOOH levels. Furthermore, Ph2Se2 prevented impairment of brain ACAP levels, as well as GPx and GST activities elicited by FB1-contaminated diets. These data suggest that dietary supplementation with 3 mg/kg Ph2Se2 prevented FB1-induced brain damage in silver catfish, and this protective effect occurred through avoided of excessive ROS production, as well as via prevention of brain lipid damage. Furthermore, Ph2Se2 exerted its neuroprotective effects via ameliorative effects on the enzymatic and non-enzymatic antioxidant defense systems, and may be an approach to prevent FB1-induced brain oxidative stress; however, is not an alternative to prevent the impairment on performance caused by FB1.

Grape pomace flour alleviates Pseudomonas aeruginosa-induced hepatic oxidative stress in grass carp by improving antioxidant defense.

Pseudomonas aeruginosa is a Gram-negative opportunistic bacterial pathogen in aquaculture systems being associated to extensive liver damage caused by oxidative stress in both marine and freshwater fish. Dietary supplementation with natural antioxidants is considered a rational strategy to prevent hepatic diseases involved with oxidative stress. Bio-residues resulting from the wine industry, such as grape pomace, are potential sources of bioactive phenolic compounds that can be applied as supplement for animal production. Thus, the aim of this study was to evaluate whether dietary supplementation with grape pomace flour (GPF) was able to prevent or reduce the hepatic oxidative damage of grass carp, Ctenopharyngodon idella, experimentally infected by P. aeruginosa. Hepatic reactive oxygen species (ROS), metabolites of nitric oxide (NOx), thiobarbituric acid reactive substances, and protein carbonylation levels were higher in fish experimentally infected by P. aeruginosa compared to the control group. Hepatic superoxide dismutase and catalase activities and antioxidant capacity against peroxyl radical levels were also higher in fish experimentally infected by P. aeruginosa compared to the control group. Dietary supplementation with 300 mg/kg GPF prevented all alterations elicited by P. aeruginosa, with the exception of protein carbonylation levels. The dietary supplementation with 150 mg/kg GPF was not able to avoid alteration of the analyzed variables, being results similar to those infected (positive control). Based on these results, dietary supplementation with 300 mg/kg GPF prevented P. aeruginosa-induced liver damage in grass carp, and this protective effect occurred through prevention on excessive ROS and NOx production, as well as via prevention of lipid damage. Moreover, 300 mg/kg GPF exerted its hepatoprotective effects by improving enzymatic and non-enzymatic antioxidant defense system. In summary, this supplementation can be an interesting approach to prevent P. aeruginosa-induced liver damage.

Oxidative stress mediated the inhibition of cerebral creatine kinase activity in silver catfish fed with aflatoxin B1-contaminated diet.

Aflatoxin B1 (AFB1) is an environmental toxicant and neurotoxic compound that induces the production of free radicals, causing oxidative stress. Creatine kinase (CK) is a central controller of energy metabolism in tissues with a large and fluctuating energy demand, and it is highly susceptible to inactivation by free radicals and oxidative damage. Thus, the aim of this study was to evaluate whether a diet for freshwater silver catfish (Rhamdia quelen) containing AFB1 inhibits cerebral CK activity, as well as the involvement of the oxidative stress on this inhibition. Brain CK activity was lower on days 14 and 21 post-feeding in animals that received AFB1-contaminated diet compared to the control group (basal diet), similarly to the brain sodium-potassium pump (Na+, K+-ATPase) activity. On the other hand, lipid peroxidation and protein carbonylation levels were higher on days 14 and 21 post-feeding in animals fed with AFB1-contaminated feed compared to the control group, while the antioxidant capacity against peroxyl radicals and thiol content was lower. Based on these evidences, the data demonstrated that diet containing AFB1 severely affects CK activity, an essential enzyme that plays an important role in brain energy homeostasis. Also, the impairment of energetic homeostasis linked with the use and generation of ATP via inhibition of CK activity elicited an inhibition of enzymes ATP-dependent, such as Na+, K+-ATPase. Moreover, the inhibition of brain CK activity appears to be mediated by the oxidation of lipids, proteins, and thiol group, as well as by a reduction in the antioxidant capacity.

Serum and hepatic oxidative damage induced by a diet contaminated with fungal mycotoxin in freshwater silver catfish Rhamdia quelen: Involvement on disease pathogenesis.

It has been recognized that oxidative stress is implicated in the initiation and progression of diseases due to the excessive formation of free radicals and impairment of the antioxidant defense system, contributing to the mortality of affected animals. The occurrence of a disequilibrium between the antioxidant/oxidant status in serum and liver of freshwater fish fed with aflatoxin B1 (AFB1) remains poorly understood and limited to only a few oxidant variables. Thus, the aim of this study was to evaluate whether an AFB1-contaminated diet causes disturbance on the antioxidant and oxidant status in silver catfish (Rhamdia quelen) of freshwater. Serum and hepatic reactive oxygen species (ROS), metabolites of nitric oxide (NOx), and lipid hydroperoxide increased on days 14 and 21 post-feeding in animals that received AFB1 contaminated diet compared to the control group (basal diet), while protein carbonylation levels increased on day 21 post-feeding. On the other hand, serum and hepatic antioxidant capacity against peroxyl radical and vitamin C levels, as well as glutathione peroxidase and catalase activities were lower on days 14 and 21 post-feeding in animals that received AFB1 contaminated diet compared to the control group. No difference was observed between groups regarding the superoxide dismutase activity and glutathione levels. Based on these evidences, an AFB1-contaminated diet causes a disturbance on serum and hepatic antioxidant/oxidant system due to lipid and protein damage elicited by excessive ROS and NOx production. Also, the antioxidant defense system was unable to avoid or minimize ROS and NOx deleterious effects, and consequently, the oxidative damage. In summary, this disturbance can contribute to understand the pathophysiology and mortality of fish after the consumption of AFB1-contaminated diets.

Aloysia triphylla in the zebrafish food: effects on physiology, behavior, and growth performance.

Dietary supplements are commonly used by animals and humans and play key roles in diverse systems, such as the immune and reproductive systems, and in metabolism. Essential oils (EOs), which are natural substances, have potential for use in food supplementation; however, their effects on organisms remain to be elucidated. Here, we examine the effects of dietary Aloysia triphylla EO supplementation on zebrafish behavior, metabolism, stress response, and growth performance. We show that fish fed diets containing A. triphylla EO presented an anxiolytic response, with reduced exploratory activity and oxygen consumption; no changes were observed in neuroendocrine stress axis functioning and growth was not impaired. Taken together, these results suggest that the A. triphylla EO supplementation is a strong candidate for use in feed, since it ensures fish welfare (anxiolytic behavior) with decreased oxygen consumption. This makes it suitable for use in high-density production systems without causing damage to the neuroendocrine stress axis and without growth performance being impaired.

Efficacy of dietary curcumin supplementation as bactericidal for silver catfish against Streptococcus agalactiae.

The antibiotics were frequently used for combating bacterial infections in aquaculture, but this treatment causes antibiotic resistance, negative impact on the environment and on health, and accumulation of residual in edible tissues. Several evidences have considered the dietary supplementation with natural products an interesting alternative to antibiotics, as the use of curcumin, a polyphenol with anti-inflammatory, antioxidant and antimicrobial properties. Thus, the aim of this study was to evaluate whether fish fed with a diet containing 150 mg curcumin/kg feed is able to enhance the resistance of silver catfish to Streptococcus agalactiae infection. Our results demonstrated that curcumin dietary supplement exerts potent bactericidal action against S. agalactiae, presenting 100% of therapeutic efficacy when compared to infected and non-supplemented animals. Also, the treatment prevented the occurrence of clinical signs of disease, as erratic swimming, corneal opacity, skin lesions in the fin and tail, and loss of appetite. In summary, curcumin can be a promising dietary supplement for improving disease resistance.

Aloysia triphylla essential oil as additive in silver catfish diet: Blood response and resistance against Aeromonas hydrophila infection.

The essential oil of Aloysia triphylla (EOAT) is a promising product with potential use in aquaculture systems. This study evaluated hematological/biochemical responses and survival of silver catfish (Rhamdia quelen) fed a diet containing EOAT and infected by Aeromonas hydrophila. After 21 days of feeding trial, fish were infected with A. hydrophila following a 10-day period of observation. Blood collection was performed before and after the bacterial challenge. Dietary EOAT by itself seems to affect some blood parameters, decreasing total leukocyte, lymphocyte, and neutrophil counts and increasing total protein values. However, 2.0 mL EOAT/kg diet showed a possible potential protective effect after A. hydrophila infection, maintaining the evaluated parameters similar to basal values (from healthy fish before the feeding trial) and promoting survival of silver catfish.

Anesthesia and anesthetic action mechanism of essential oils of Aloysia triphylla and Cymbopogon flexuosus in silver catfish (Rhamdia quelen).

OBJECTIVES: To document the time for anesthesia induction and recovery using different concentrations of essential oils (EOs) of Cymbopogon flexuosus and Aloysia triphylla in silver catfish (Rhamdia quelen), and to determine whether the mechanism of action of either EO involves the benzodiazepine (BDZ) site of the GABAA receptor. STUDY DESIGN: Experimental study. ANIMALS: A total of 144 silver catfish, length 7.5 ± 1.1 cm, weighing 3.95 ± 0.85 g. METHODS: Essential oils were evaluated at concentrations of 25, 150 and 300 µL L-1, and also ethanol alone (seven groups, n = 6 per group). Induction of sedation or anesthesia and recovery were assessed. In a further six groups (n = 6 per group), fish were exposed to both EOs (25, 150 or 300 µL L-1) with diazepam 150 µm, and also diazepam (10 µm) alone. Flumazenil (5 or 10 µm) was added to the recovery water of fish exposed to diazepam (150 µm) or both EOs (150 and 300 µL L-1) (total of 10 groups = 60 fish). RESULTS: Both EOs induced anesthesia at concentrations of 150 and 300 µL L-1, and sedation at 25 µL L-1. There was no significant difference between EOs for reaching deep anesthesia; there was a significantly longer recovery time for the EO of C. flexuosus. The addition of diazepam (150 µm) resulted in faster induction of anesthesia with both EOs, with no significant change in recovery times. Flumazenil (10 µm) reversed the diazepam-induced anesthesia, but not the anesthesia induced by EOs. CONCLUSIONS AND CLINICAL RELEVANCE: The EO of C. flexuosus induced effective sedation (25 µL L-1) and anesthesia (150 and 300 µL L-1) without short-term mortality. The modulation of the BDZ site of the GABAA receptor in the anesthetic action mechanism of both EOs was not demonstrated.

Pre-sedation and transport of Rhamdia quelen in water containing essential oil of Lippia alba: metabolic and physiological responses.

The effects of transporting silver catfish (Rhamdia quelen) for 6 h in plastic bags containing 0 (control), 30 or 40 µL/L of essential oil (EO) from Lippia alba leaves were investigated. Prior to transport, the fish in the two experimental groups were sedated with 200 µL/L of EO for 3 min. After transport, dissolved oxygen, carbon dioxide, alkalinity, water hardness, pH, temperature and un-ionized ammonia levels in the transport water did not differ significantly among the groups. However, total ammonia nitrogen levels and net Na(+), Cl(-) and K(+) effluxes were significantly lower in the groups transported with EO of L. alba than those in the control group. PvO2, PvCO2 and HCO3(-) were higher after transporting fish in 40 µL/L of EO of L. alba, but there were no significant differences between groups regarding blood pH or hematocrit. Cortisol levels were significantly higher in fish transported in 30 µL/L of EO of L. alba compared to those of the control group. The metabolic parameters (glycogen, lactate, total amino acid, total ammonia and total protein) showed different responses after adding EO to the transport water. In conclusion, while the EO of L. alba is recommended for fish transport in the conditions tested in the present study because it was effective in reducing waterborne total ammonia levels and net ion loss, the higher hepatic oxidative stress in this species with the same EO concentrations reported by a previous study led us to conclude that the 10-20 µL/L concentration range of EO and lack of pre-sedation before transport are more effective.

Lippia alba essential oil promotes survival of silver catfish (Rhamdia quelen) infected with Aeromonas sp.

In vitro and in vivo activity of the Lippia alba essential oil (EO) against Aeromonas sp. was evaluated. In the in vitro assay the minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) of EO for Aeromonas cells were determined using the microdilution method. Twenty five strains of Aeromonas sp. isolated from infected fish obtained from local fish farms were used. MIC and MBC values were 2862 and 5998 µg mL-1 for L. alba EO and 0.5 and 1.2 µg mL-1 for gentamicin, respectively. In the in vivo assay silver catfish juveniles (Rhamdia quelen) (7.50 ± 1.85 g and 10.0 ± 1.0 cm) with typical injuries associated to Aeromonas infection were divided into four treatments (in triplicate n=10): untreated fish (negative control), 10 mg L-1 of gentamicin, and 20 or 50 µL L-1 of EO. Fish were maintained in aerated 20 L plastic boxes. After 10 days survival of silver catfish infected with Aermonas sp. and treated with essential oil (50 µL L-1) was greater than 90%.

Using the essential oil of Aloysia triphylla (L'Her.) Britton to sedate silver catfish (Rhamdia quelen) during transport improved the chemical and sensory qualities of the fish during storage in ice.

UNLABELLED: Exposure of silver catfish to 40 µL/L of the essential oil of Aloysia triphylla (AT) during in vivo transport delayed the onset and resolution of rigor mortis as well as the degradation of IMP into HxR compared to the control. The fish that were treated with 30 or 40 µL/L of AT received lower sensory demerit scores after 10 d of storage in ice compared to the control, and the fish that were treated with 40 µL/L of AT had a longer sensory shelf life than did the control. These results indicated that using AT as a sedative in the water in which the silver catfish were transported extended their freshness and increased their shelf life during refrigerated storage. PRACTICAL APPLICATION: Interest in natural anesthetics, such as Aloysia triphylla, has increased in the field of commercial aquaculture because they reduce the number of fish lesions acquired during capture, handling, and transportation. Fish sedated with the essential oil of A. triphylla at 40 µL/L during transport before slaughter exhibited a delay in the loss of freshness and an increased shelf life in ice. In addition to improving animal welfare before slaughter, the essential oil appears to be a promising product for improving fish conservation in the food industry.

Anesthetic activity of the essential oil of Aloysia triphylla and effectiveness in reducing stress during transport of albino and gray strains of silver catfish, Rhamdia quelen.

This study investigated the efficacy of the essential oil (EO) of Aloysia triphylla as an anesthetic for albino and gray strains of silver catfish, Rhamdia quelen. Juveniles were exposed to concentrations between 20 and 800 µL L(-1) EO of A. triphylla to evaluate time of induction and recovery from anesthesia. In another experiment, both strains were divided into four groups such as 0 (control), 30, 40, or 50 µL L(-1) EO and transported for 5 h. The longest time for anesthetic induction and recovery was observed in the albinos. Both strains reached anesthesia in the 100-800 µL L(-1) (11.1-1.24 min) range, without mortality, being 200 µL L(-1) the best response considering time to anesthesia (5.35 min). Albinos transported with all EO concentrations showed higher values of carbon dioxide in the water of transport, but lower levels were observed in grays transported with 40 and 50 µL L(-1) EO when compared to control fish. The same concentrations did not prevent significant whole-body cortisol rise at the end of transport in the albino strain. Juveniles of both strains transported with EO presented lower ion loss to the water compared to control fish. The EO of A. triphylla is an effective anesthetic for albino and gray silver catfish. This EO increases whole-body cortisol levels in the albino strain, but as it reduces net ion loss as in the gray strain, it can be also recommended for transport.

The use of nitazoxanide against the pathogens Ichthyophthirius multifiliis and Aeromonas hydrophila in silver catfish (Rhamdia quelen).

The antiparasitic and antibacterial activities of nitazoxanide (NTZ) against Ichthyophthirius multifiliis (Ich) and Aeromonas hydrophila, respectively, were evaluated in silver catfish juveniles (Rhamdia quelen). In the first experiment, fish naturally infected by Ich were treated with different concentrations of NTZ. Fish mortality and the number of trophonts/juveniles were evaluated. In the second experiment, the antimicrobial potential in vitro (MIC test) and preventive potential in vivo against A. hydrophila was evaluated. The treatment with 1.5mgL(-1) NTZ showed the best result against Ich, with a significant reduction of trophonts and fish mortality of 10%. NTZ did not show in vitro activity against A. hydrophila at concentrations tested, but showed a probable preventive activity in vivo. However, these results are preliminary and more studies should be conducted.

Transportation of silver catfish, Rhamdia quelen, in water with eugenol and the essential oil of Lippia alba.

This study investigated the effectiveness of eugenol and of the essential oil (EO) of Lippia alba when used in the transport of the silver catfish (Rhamdia quelen). These investigations involved measurements of blood (pH, PvO(2), PvCO(2) and HCO(3) (-)) and water parameters, survival and ionoregulatory balance. Fish (301.24 ± 21.40 g, 28.90 ± 1.30 cm) were transported at a loading density of 169.2 g L(-1) for 4 h in fifteen plastic bags (7 L) divided into five treatments: control, 1.5 or 3.0 µL L(-1) of eugenol and 10 or 20 µL L(-1) of EO of L. alba. The water parameters were measured before (0 h) and after (4 h) transportation. The net Na(+), Cl(-) and K(+) losses were higher in fish from the control treatment compared to the other treatments. The PvO(2), PvCO(2) and HCO(3) (-) increased significantly in all of the treatments at the end of the transport period. In conclusion, based on the water (total ammonia nitrogen) and ionoregulatory indicators determined in the present study, our findings indicate that eugenol and the EO of L. alba are recommended for use in the transport of this species because these anesthetics apparently reduce stress.