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The potential of bitter orange peel powder (BOPP) as a nutritional strategy for fish was investigated in Nile tilapia. A total of 120 juveniles with an average initial weight of 9.8 ± 0.7 g were divided into four groups, replicated three times, resulting in 12 experimental units (60 L each) at a stocking density of 1.63 g of fish per liter. Productive parameters, whole-body composition, blood biochemistry, erythroid morphometry, intestinal histology, and heat tolerance were assessed in the juveniles subjected to one of the following treatments: non-supplemented basal diet (control group); basal diet with BOPP at 10 g/kg (BOPP10 group); basal diet with BOPP at 20 g/kg (BOPP20 group); and basal diet with BOPP at 40 g/kg (BOPP40 group). The BOPP additive had a positive influence on Nile tilapia growth, as final weight and weight gain were greater in all BOPP-treated fish, despite the reduction in crude protein in BOPP10 and BOPP20 groups. Fish receiving BOPP40 had an increase in total lipids and showed the highest levels of triglycerides and total cholesterol. Villi development was greater in the tilapia given BOPP10. It may be concluded that BOPP presented the most promising results for Nile tilapia juveniles when used at 10 g/kg diet. Regarding the erythroid morphometry, there was a general increase in nuclear and cytoplasmic areas in BOPP-fed tilapia; this seems to be the first report on the direct impact of the inclusion of functional additives in fish diet upon such parameters. As concerns the thermal tolerance evaluated at the end of the feeding trial, no differences were registered among the experimental groups. Thus, BOPP represents a feasible alternative ingredient to be explored in fish nutrition, since orange peel is a natural low-cost source of essential nutrients and valuable bioactive compounds., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

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/m 3 ) and fed (HSD-F), high stocking density and fasted (HSD-FS), low stocking density (2.5 kg/m 3 ) 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.

It is well documented today that pesticides, used in crop production, may modulate the immune system of healthy fish. However, there is still only limited information regarding the effects of these anthropogenic stressors in conjunction with natural stressors (pathogens), on the innate immune responses of freshwater fish. Thus, the aim of this investigation was to compare the innate immune response of two groups of fish (Rhamdia quelen), naturally infected with Aeromonas hydrophila, exposed and unexposed to a non-lethal concentration of chlorantraniliprole (CAP) insecticide (0.0 and 1.3 μg/L/24 h). Unhealthy fish exposed to CAP showed significant higher total leukocyte counts and neutrophils percentage compared to non-exposed infected fish). However, the monocytes and eosinophils percentage significantly decreased in fish exposed to CAP. Furthermore, lysozyme activity values measured in plasma, skin mucus, gill and intestine significantly reduced in fish exposed to CAP. The CAP-induced immunomodulation may interfere on the ability of the animal to heal or fight the infection, and possible contribute to the spread of bacterial infection in fish production or environment., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Algamune ™ is a commercial additive produced from Euglena gracilis, providing a rich source of the β-1,3-glucan paramylon. Isolated kidney phagocytes of Nile tilapia were incubated with graded doses (0, 8, 16, 32, 64 and 128 μg/ml) of Algamune ™ and purified paramylon to gauge their ability to elicit the production of reactive oxygen species. A linear response was observed for extracellular superoxide anion for both sources but only Algamune ™ for intracellular superoxide anion. After corroborating the immunostimulant properties ex vivo, a feeding trial was conducted to evaluate the dietary supplementation of Algamune ™ (0, 100, 200, 400 and 800 mg/kg of diet) for Nile tilapia. Fish were fed for 3 weeks, after which, fish were sampled for blood and head kidney phagocytes. The remaining fish were challenged with Streptococcus iniae. Macrophage extracellular superoxide anion production was significantly elevated in fish fed diets with 200 mg of Algamune ™ kg -1 when compared to fish fed the basal diet. Even though the disease challenge did not show statistical differences, it is worth mentioning that fish fed intermediate doses of Algamune ™ had lowest numerical mortality values. Therefore, Algamune ™ was demonstrated to enhance some immunological responses of tilapia both in ex vivo and in vivo., (© 2018 John Wiley & Sons Ltd.)

Aims: This study investigated the antibacterial activity of five phytochemicals (carvacrol, citral, eugenol, linalool and thymol) alone or in combination with florfenicol or oxytetracycline against bacteria isolated from silver catfish (Rhamdia quelen). We also analysed the potential of these compounds to inhibit biofilm formation and haemolysis caused by the bacteria., Methods and Results: Bacteria were tested with antimicrobials to calculate the multiple antibiotic resistances. The checkerboard assay was used to evaluate a putative synergy between five phytochemicals and antimicrobials against the strains isolated. The biofilm formation inhibition assay was performed with phytochemicals and antimicrobials, and the haemolysis inhibition assay was performed with the phytochemicals. Carvacrol, eugenol and thymol were the most effective phytochemicals. Three combinations (linalool with florfenicol or oxytetracycline against Aeromonas hydrophila and citral with oxytetracycline against Citrobacter freundii) demonstrated synergy in the checkerboard assay. All phytochemicals inhibited biofilm formation and haemolysis activity., Conclusion: The tested phytochemicals showed satisfactory activity against fish pathogenic bacteria., Significance and Impact of the Study: The phytochemicals did not present antagonistic interactions with the antimicrobials, allowing their combined use, which may contribute to a decrease in the use of conventional drugs and their residues in aquatic environment., (© 2018 The Society for Applied Microbiology.)

This study evaluated the activity of leaf essential oil (EO) from Hyptis mutabilis as well as its major constituent, (-)-globulol, in infections by the parasite Ichthyophthirius multifiliis (ich). Effects on hematological, biochemical, and immunological parameters of silver catfish, Rhamdia quelen, exposed to the same samples also were evaluated. In the first experiment, naturally infected fish were treated with EO (0, 10, and 20 mg · L -1 ) and ethanol, using several methods of exposure. Fish mortality and the number of trophonts per fish were assessed after 48 and 96 hr. Hour-long daily baths resulted in optimal survival, so this methodology was used for the second experiment, in which infected animals were exposed to (-)-globulol at 2.5 and 5 mg · L -1 . The most effective concentrations in Experiments 1 and 2 were chosen for Experiment 3, in which healthy animals were subjected to hour-long daily baths with EO (20 mg · L -1 ) or (-)-globulol (2.5 mg · L -1 ). Additionally, an in vitro experiment was performed with EO and globulol at the same concentrations of the in vivo test. EO and (-)-globulol increased the survival of fish infected with ich and altered certain hematological and biochemical parameters. After 4 days, levels of hematocrit, erythrocytes, and leukocytes increased significantly in healthy animals exposed to EO. Exposure to (-)-globulol increased leukocyte number alone. No significant differences in nonspecific immunological parameters were detected when treated groups were compared to controls, but the leukocytosis observed in EO- and globulol-treated healthy animals indicates that EO and (-)-globulol increased innate immunity in these fish. An in vitro antiparasitic effect was observed in both samples.

The effect of dissolved organic carbon (DOC) against the burden of low pH was assessed in silver catfish Rhamdia quelen through the gills. A commercial humic acid (HA) was used as the source of DOC at 0 (control), 10, 25, and 50 mg/l. For each HA concentration, two pH levels were tested: 6.5 (control) and 5.5. After 40 days of exposure, the gills were removed and morphological variables were analyzed through light and scanning electronic microscopy. The low water pH caused a reduction in the length of filaments, number of lamellae, and gill respiratory surface area (GRSA) and an increase in chloride cells (CCs) number in the filament epithelium. When HA was added to the test water, GRSA increased and CCs proliferated in lamellae, suggesting ionic and respiratory disturbances. Scanning electronic microscopy revealed that CC morphometric variables, that is, apical area, fractional area, and density, were undisturbed by the pH reduction, but increased in the presence of HA. This study indicates that the commercial HA did not protect silver catfish against low pH stress. Instead, it caused changes that may affect vital processes such as ion regulation and ventilation and, consequently, reduce growth., (© 2017 Wiley Periodicals, Inc.)

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., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Linalool is the main compound of many essential oils and occurs in two isomeric forms: S-(+)- and R-(-)-linalool. This study aimed to determine if linalool isomers have different antimicrobial and anesthetic properties in fish. For this purpose, these compounds were previously isolated from Lippia alba (Mill.)N. E. Brown and Ocimum americanum L. essential oils. Antimicrobial effects were evaluated through the microdilution test against Aeromonas hydrophila, an important fish disease etiologic agent. Induction time until sedation, anesthesia and recovery time were determined in silver catfish (Rhamdia quelen) through bath exposure (60, 180, 300 or 500 μL L-1). The results showed different biological properties for the isomers being S-(+)-linalool the only active against A. hydrophila at 3.2 mg mL-1. The sedation was induced without differences between the compounds, however R-(-)-linalool promoted faster anesthesia. There were no differences regarding the recovery time of the animals exposed to the linalool isomers. Although both S-(+)- and R-(-)-linalool can be used for sedative purposes, their use in A. hydrophila infection is inadvisable due to the high effective concentration. Considering anesthesia as the main objective, the R-(-)-linalool demonstrated clear advantages at lower concentration.

Phytochemicals such as plant essential oils (EOs) have been reported to favour various activities in the innate immune system of fish. Thus, the aim of this study was to verify the in vitro effect of three different plant EOs (Ocimum americanum, Cymbopogon flexuosus and Melaleuca alternifolia) on non-specific immune parameters and erythrocyte osmotic fragility of red drum, Sciaenops ocellatus. Concentrations of each plant EO evaluated in preparations of head-kidney macrophages, blood leucocytes and blood plasma were as follows: 0.0 (control), 1.0, 2.0, 4.0, 8.0, and 16.0 μg/ml. Red drum head-kidney macrophages significantly increased extracellular superoxide anion production when exposed (20 h) to O. americanum EO (1.0-8.0 μg/ml) and C. flexuosus EO (2.0 and 4.0 μg/ml). The respiratory burst of blood leucocytes (NBT test) significantly increased in all concentrations when compared to the respective control group, for all EOs. At the highest concentration (16.0 μg/ml), C. flexuosus EO significantly inhibited the haemolytic activity of complement system in red drum blood after 1 h exposure. None of the tested concentrations significantly altered plasma lysozyme activity or erythrocyte osmotic fragility after exposing (1 h) red drum whole blood to each EO. This study demonstrated that these plant EOs are capable of triggering superoxide anion production in red drum leucocytes (head-kidney macrophages and/or blood leucocytes). In vivo studies are warranted to address their potential as immunostimulants in the diet of red drum and other aquacultured species., (Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.)

This study evaluated productive parameters, whole-body composition, non-specific immune responses and pH and microbiota of digestive tract contents of red drum (Sciaenops ocellatus) fed diets supplemented with Ocimum americanum essential oil (OAEO) (0 - control, 0.25, 0.5, 1.0 and 2.0 g/kg diet). After 7 weeks no significant differences in productive parameters and whole-body composition were observed. Plasma and intestinal lysozyme measurements and pH of the stomach and intestine (6 h after feeding) did not show significant differences among groups. Intestinal microbial community in fish fed the basal and OAEO diets (all concentrations) were identical. However, red drum fed the diet with OAEO at 1.0 g/kg had significantly increased intraperitoneal fat deposition and stomach pH (2 h after feeding) and decreased superoxide ion production (NBT-test) compared to the control group. Hemolytic activity of the complement system increased in fish fed diets containing OAEO. Red blood cells from fish fed the lowest OAEO concentration (0.25 g/kg) showed significant lower fragility in erythrocyte osmotic fragility assay, but fish fed 0.5 and 1.0 g/kg showed significant higher erythrocyte fragility. Lysozyme measurement in the supernatant of stomach content was significantly higher in fish fed the diet supplemented at 0.5 g/kg. Based on these various results, OAEO at different supplementation levels did not influence growth performance and intestinal microbial community; however, the EO added to the diet showed effects on immunological responses of red drum., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Unlabelled: The bactericidal activity (minimum inhibitory concentration (MIC)-test) of Ocimum americanum (inflorescences) essential oil (OAEO) against Aeromonas hydrophila was determined in this study. Also investigated was the potential of OAEO and the main compound found in the oil (linalool) at subinhibitory concentrations to be inhibitors of haemolysis caused by Aer. hydrophila in fish erythrocytes. An in vivo experiment was conducted to evaluate the survival of fish (Rhamdia quelen) experimentally infected with Aer. hydrophila and exposed to OAEO. A second experiment was conducted to evaluate the in vitro and in vivo activity of OAEO (mix from inflorescences and leaves) against the parasite Gyrodactylus sp. The OAEO showed weak in vitro activity against Aer. hydrophila (6400 μg ml(-1) ). Subinhibitory concentrations of OAEO (100 μg ml(-1) ) inhibited haemolysis (90%) caused by Aer. hydrophila in fish erythrocytes, however, linalool did not inhibit haemolysis activity. At the low concentrations (10 and 20 mg l(-1) ) added to the water, OAEO promoted the survival of fish experimentally infected with Aer. hydrophila. Lastly, the OAEO mix (50 mg l(-1) ) was effective against Gyrodactylus sp., significantly reducing (60%) the number of parasites in the fish., Significance and Impact of the Study: Phytochemicals, such as essential oils (EOs) are a great source of new molecules and have shown potential to be used in aquaculture systems. However, additional studies focused on the in vivo efficacy, mode of action and identification of the active compounds are needed. This study determined the potential of Ocimum americanum EO for use against two important fish pathogens, Aeromonas hydrophila and Gyrodactylus sp., as well as providing preliminary information about the role of the main EO compound (linalool) against Aer. hydrophila virulence., (© 2016 The Society for Applied Microbiology.)

Imazapyr (IMY) and imazapic (IMI) are imidazolinone herbicides which have been associated in a commercial formulation (Kifix(®)). To date, there are no studies on the toxicity of an IMY+IMI herbicide in fish. This work aimed to assess the acute toxicity (24 and 96 h) of IMY+IMI (0, 0.488 and 4.88 µg/L) towards Rhamdia quelen through hematological, biochemical, immunological, ionoregulatory and enzymatic indexes. Red blood cell count was lower at 4.88 than at 0.488 µg/L (24 and 96 h); mean corpuscular volume was lower than control at both concentrations (24 h) and at 0.488 µg/L (96 h); lymphocytes declined at 4.88 µg/L comparing to control (96 h); and monocytes increased at 4.88 µg/L (96 h) in comparison with the respective control and with 4.88 µg/L at 24h. Aspartate aminotransferase was higher at 0.488 µg/L (96 h) than the respective control and the respective concentration at 24 h; uric acid reduced at 4.88 µg/L comparing with 0.488 µg/L (96 h); and cortisol was lower at 4.88 µg/L compared to 0.488 µg/L and control (96 h). Herbicide exposure lowered plasma bactericidal activity at both concentrations (24 h) and at 0.488 µg/L (96 h); and plasma complement activity declined at 4.88 µg/L comparing with 0.488 µg/L and control (96 h), and was lower at all concentrations at 96 h than at 24 h. Plasma K(+) levels were higher at 4.88µg/L than in the remaining groups (24 and 96h); and Na(+) levels decreased at 4.88 µg/L compared to control (96 h). Na(+)/K(+)-ATPase and H(+)-ATPase activities in gills were lower at 4.88 µg/L comparing with control (24 h) and with the respective concentration at 96 h; and AChE activity in brain was higher at 0.488 and 4.88 µg/L than control (24 h) and the respective concentrations at 96 h, while in muscle it was higher at 0.488 and 4.88 µg/L than control (96 h) and the respective concentrations at 24 h. The present findings demonstrate that, despite IMY+IMI targets the animal-absent AHAS enzyme, such formulation displayed an acute toxic effect upon R. quelen homeostasis by impacting on vital functions such as immune defense, metabolism, ionoregulation and neurotransmission., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Aims: The aims of this study were to investigate the in vitro antibacterial activity of the essential oils (EOs) of Hesperozygis ringens (HREO), popularly known as 'espanta-pulga' and two different species of basil, Ocimum gratissimum (OGEO) and Ocimum americanum (OAEO), as well as, the potential of these products to be used in silver catfish (Rhamdia quelen) infected with Aeromonas hydrophila., Methods and Results: OGEO and HREO showed better antibacterial activity in vitro. Subinhibitory concentrations of all EOs inhibited haemolysis caused by Aer. hydrophila in fish erythrocytes (100% reduction for OAEO at 100 μg ml(-1) and more than 90% for HREO and OGEO at 150 μg ml(-1) ). However, OAEO and HREO showed the best survival results (75 and 70% respectively) after their use as treatment (therapeutic baths-1 h daily/5 days) in silver catfish experimentally infected with Aer. hydrophila. A second in vivo assay using healthy fish was conducted to verify the potential of the EOs (preventive baths-1 h daily/5 days) to promote fish survival. Fish exposed to HREO and OAEO and their diluent (ethanol) showed significant lower haematocrit values and higher complement system activity compared to control. Plasma cortisol level was significantly higher in the groups exposed to both EOs. There was no significant difference in survival of silver catfish challenged with Aer. hydrophila after preventive baths with HREO, OAEO and control group., Conclusions: All tested EOs showed in vitro antibacterial properties against Aer. hydrophila and HREO and OAEO showed potential to be used in the treatment of infected fish., Significance and Impact of the Study: These products can be used in aquaculture as therapeutic and prophylactic agents against fish pathogens, with antimicrobial and/or immunostimulant properties., (© 2015 The Society for Applied Microbiology.)

Rhamdia quelen morphophysiological responses to propofol sedation were examined. The purpose was to investigate whether propofol would be a suitable drug to be used in fish transport procedures. Fish were exposed to 0, 0.4 or 0.8 mg L(-1) propofol for 1, 6 or 12 h in 40 L tanks, simulating open transport systems. Propofol was able to prevent the peak of cortisol levels experienced by the group exposed to 0 mg L(-1) propofol at 1 h. At 0.4 mg L(-1), propofol also preserved the stability of hematological (hematocrit, red blood cell count, hemoglobin, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration), morphological (red blood cell area), biochemical (cortisol, glucose, lactate, total protein, ammonia, urea, alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase) and hydromineral (Na(+), Cl(-) and K(+) plasma levels) indicators of stress. Such results suggest that sedation with propofol at 0.4 mg L(-1) is suitable for R. quelen transport.

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%.

The aim of this study was to evaluate the activity of eugenol against the fish pathogen Aeromonas hydrophila and eugenol's effect on hematological and natural immune parameters in silver catfish (Rhamdia quelen). In vitro, eugenol showed weak activity against A. hydrophila, but in vivo, at a subinhibitory concentration (10 mg L(-1)), it promoted survival in infected silver catfish. Eugenol (50 μg mL(-1)) reduced the hemolytic activity of A. hydrophila supernatant in vitro in fish erythrocytes. Subjecting catfish to eugenol baths (5 and 10 mg L(-1)) for five days did not alter the hematological and immunological parameters studied in this work. Based on these results, eugenol can be used to treat or prevent bacterial diseases in fish., (Copyright © 2013 Elsevier B.V. All rights reserved.)

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., (Copyright © 2013 Elsevier B.V. All rights reserved.)