Dietary addition of rutin impairs inflammatory response and protects muscle of silver catfish (Rhamdia quelen) from apoptosis and oxidative stress in Aeromonas hydrophila-induced infection.

This research aimed to assess the influence of dietary addition of rutin on inflammation, apoptosis and antioxidative responses in muscle of silver catfish (Rhamdia quelen) challenged with Aeromonas hydrophila (A. hydrophila). Fish were split into four groups as follows: control, 0.15% rutin, A. hydrophila, 0.15% rutin + A. hydrophila. After 2 weeks of feeding with standard or rutin diets, fish were challenged or not with A. hydrophila for 1 week. Rutin-added diet abrogates A. hydrophila induced-hemorrhage and inflammatory infiltration. It decreases A. hydrophila induced-apoptosis through decreasing the ratio of Bax to Bcl-2 and increasing phospho-Akt to Akt ratio. It diminishes the A. hydrophila induced-rise in nitric oxide and superoxide anion levels and reestablishes superoxide dismutase activity as well. Although such diet is unable to recover the levels of reduced glutathione (GSH), cysteine and glutamate cysteine ligase, which are depleted as a result of A. hydrophila infection, it diminishes the oxidized glutathione (GSSG) content, thus decreasing GSSG to GSH ratio. It increases the levels of cysteine residues of proteins and diminishes those of thiol-protein mixed disulfides, which were changed after A. hydrophila challenge. Finally, it reduces A. hydrophila induced-lipid peroxidation, markedly elevates ascorbic acid and thus reestablishes total antioxidant capacity, whose levels were decreased after A. hydrophila challenge. In conclusion, the dietary addition of rutin at 0.15% impairs A. hydrophila-induced inflammatory response, inhibits A. hydrophila-induced apoptosis and promotes cell survival. It also reduces the A. hydrophila-induced oxidative stress and stimulates the antioxidative responses in muscle of A. hydrophila-infected silver catfish.

Citrobacter freundii impairs the phosphoryl transfer network in the gills of Rhamdia quelen: Impairment of bioenergetics homeostasis.

The precise coupling of spatially separated intracellular adenosine triphosphate (ATP)-producing and ATP-consuming, catalyzed by creatine kinase (CK), adenylate kinase (AK), and pyruvate kinase (PK), is a critical process in the bioenergetics of tissues with high energy demand, such as the branchial tissue. The effects of Citrobacter freundii infection on gills remain poorly understood, limited only to histopathological studies. Thus, the aim of this study was to evaluate whether experimental infection by C. freundii impairs the enzymes of the phosphoryl transfer network in gills of silver catfish (Rhamdia quelen). The CK (cytosolic and mitochondrial) and AK activities decreased in infected compared to uninfected animals, while the PK activity did not differ between groups. The gill histopathology of infected animals revealed extensive degeneration with fusion and necrosis of secondary lamellae, detachment of superficial epithelium, aneurysm, vessel congestion and inflammatory process. Based on these evidences, the inhibition and absence of an efficient communication between CK compartments caused the impairment of the branchial bioenergetics homeostasis, which was not compensated by the augmentation on branchial AK activity in an attempt to restore energy homeostasis. In summary, these alterations contribute to disease pathogenesis linked to branchial tissue in animals infected with C. freundii.

Aeromonas caviae alters the cytosolic and mitochondrial creatine kinase activities in experimentally infected silver catfish: Impairment on renal bioenergetics.

Cytosolic and mitochondrial creatine kinases (CK), through the creatine kinase-phosphocreatine (CK/PCr) system, provide a temporal and spatial energy buffer to maintain cellular energy homeostasis. However, the effects of bacterial infections on the kidney remain poorly understood and are limited only to histopathological analyses. Thus, the aim of this study was to investigate the involvement of cytosolic and mitochondrial CK activities in renal energetic homeostasis in silver catfish experimentally infected with Aeromonas caviae. Cytosolic CK activity decreased in infected animals, while mitochondrial CK activity increased compared to uninfected animals. Moreover, the activity of the sodium-potassium pump (Na+, K+-ATPase) decreased in infected animals compared to uninfected animals. Based on this evidence, it can be concluded that the inhibition of cytosolic CK activity by A. caviae causes an impairment on renal energy homeostasis through the depletion of adenosine triphosphate (ATP) levels. This contributes to the inhibition of Na+, K+-ATPase activity, although the mitochondrial CK activity acted in an attempt to restore the cytosolic ATP levels through a feedback mechanism. In summary, A. caviae infection causes a severe energetic imbalance in infected silver catfish, which may contribute to disease pathogenesis.

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.

Melaleuca alternifolia essential oil enhances the non-specific immune system and prevents oxidative damage in Rhamdia quelen experimentally infected by Aeromonas hydrophila: Effects on cholinergic and purinergic systems in liver tissue.

The aim of this study was to evaluate the effects of M. alternifolia essential oil used to treat silver catfish (Rhamdia quelen) experimentally infected by Aeromonas hydrophila on oxidative stress variables, and for the first time, on hepatic enzymes of the cholinergic and adenosinergic systems. For that, fish were divided into six groups (A-F), each containing seven animals. Groups A, B and C were composed of uninfected animals, while animals in groups D, E and F were intramuscularly inoculated with A. hydrophila. Groups B and E received a prophylactic bath with M. alternifolia essential oil (50 µL/L, diluted in ethanol) for seven days, while groups C and F were exposed to ethanol. After the prophylactic baths, groups D, E and F were inoculated with 100 µL of A. hydrophila solution (2.1 × 109 colony-forming unit). Two days after inoculation, the animals were euthanized and liver samples were collected. Infected animals (the group D) showed increased TBARS and protein carbonylation levels, while CAT, AChE and ADA activities decreased compared to uninfected animals (the group A). The prophylactic treatment with M. alternifolia essential oil (the group E) prevented the alterations caused by A. hydrophila, but it did not change AChE activity. Thus, the prophylactic treatment prevents damage caused by lipids and proteins, as well as alterations of the adenosinergic system, demonstrating that the anti-inflammatory effect of TTO is mediated by the adenosinergic pathway. In addition, TTO prophylactic treatment might be considered an important approach to prevent the hepatic damage caused by A. hydrophila.

Involvement of cholinergic and purinergic systems during the inflammatory response caused by Aeromonas hydrophila in Rhamdia quelen.

The aim of this study was to evaluate the cholinergic (acetylcholinesterase (AChE) and butyrylcholinesterase (BChE)) and purinergic (adenosine deaminase (ADA)) systems in head kidney, spleen, total blood and serum samples in experimentally infected fish with A. hydrophila, and the involvement of these systems during the inflammatory process. Silver catfish (Rhamdia quelen) juveniles were divided into two groups with seven fish each: uninfected (negative control) and infected (positive control). On day 2 post-infection, animals were euthanized and the head kidney, spleen, total blood and serum were collected. AChE and ADA activities in head kidney and spleen decreased in infected animals compared to uninfected animals, as well as AChE in total blood and seric ADA activities. BChE activity was not expressed in the evaluated tissues. Therefore, our results lead to the hypothesis that cholinergic and purinergic systems play an important role on the immune response against A. hydrophila with an anti-inflammatory effect. In summary, AChE and ADA activities reduced probably in order to protect against tissue inflammatory damage caused by infection.