Branchial bioenergetics dysfunction as a relevant pathophysiological mechanism in freshwater silver catfish (Rhamdia quelen) experimentally infected with Flavobacterium columnare.

Flavobacterium columnare, the causative agent of columnaris disease, is a serious bacterial disease responsible for causing devastating mortality rates in several species of freshwater fish, leading to severe economic losses in the aquaculture industry. Notwithstanding the enormous impacts this disease can have, very little is known regarding the interaction between the host and bacterium in terms of the mortality rate of silver catfish (Rhamdia quelen), as well its linkage to gill energetic homeostasis. Therefore, we conducted independent experiments to evaluate the mortality rates caused by F. columnare in silver catfish, as well as whether columnaris disease impairs the enzymes of the phosphoryl transfer network in gills of silver catfish and the pathways involved in this inhibition. Experiment I revealed that clinical signs started to appear 72 h post-infection (hpi), manifesting as lethargy, skin necrosis, fin erosion and gill discoloration. Silver catfish began to die at 96 hpi, and 100% mortality was observed at 120 hpi. Experiment II revealed that creatine kinase (CK, cytosolic and mitochondrial) and pyruvate kinase (PK) activities were inhibited in silver catfish experimentally infected with F. columnare, while no significant difference was observed between experimental and control groups with respect to adenylate kinase activity. Activity of the branchial sodium-potassium pump (Na+, K+-ATPase) was inhibited while reactive oxygen species (ROS) and lipid peroxidation levels were higher in silver catfish experimentally infected with F. columnare than in the control group at 72 hpi. Based on these data, the impairment of CK activity elicited by F. columnare caused a disruption in branchial energetic balance, possibly reducing ATP availability in the gills and provoking impairment of Na+, K +ATPase activity. The inhibition of CK and PK activities appears to be mediated by ROS overproduction and lipid peroxidation, both of which contribute to disease pathogenesis associated with branchial tissue.

Caffeine supplementation in diet mitigates Aeromonas hydrophila-induced impairment of the gill phosphotransfer network in grass carp Ctenopharyngodon idella.

Some evidence suggests the involvement of phosphotransfer network in the pathogenesis of fish bacterial diseases, catalyzed by creatine kinase (CK), pyruvate kinase (PK) and adenylate kinase (AK); nevertheless, the effects on fish affected by Aeromonas hydrophila remain unknown. Recent evidence suggested a potent protective effect of caffeine on the branchial phosphotransfer network of fish subjected to challenge conditions. Therefore, the aim of this study was to evaluate whether A. hydrophila infection impaired branchial bioenergetics. We also determined whether dietary supplementation with caffeine protected against A. hydrophila-induced gill bioenergetic imbalance. We found that branchial cytosolic CK and AK activities were significant lower in fish experimentally infected with A. hydrophila than in uninfected fish, while mitochondrial CK activity was significant higher. Branchial lactate dehydrogenase (LDH) activity and lactate levels were significant higher in fish experimentally infected by A. hydrophila than in uninfected fish, while sodium-potassium ion pump (Na+, K+-ATPase) activity and adenosine triphosphate (ATP) levels were significant lower. No significant difference was observed between groups with respect to branchial PK activity. The dietary supplementation with 8% caffeine improved the branchial CK (cytosolic and mitochondrial), AK, and LDH activities, as well as ATP levels, but did not prevent increases in branchial lactate levels or the inhibition of Na+, K+-ATPase activity elicited by aeromonosis. Based on this evidence, we believe that reduction of CK (cytosolic) and AK activities contributes to impairment of bioenergetic homeostasis, while augmentation of mitochondrial CK activity can be considered an attempt to prevent or reduce the energetic imbalance during aeromonosis caused by A. hydrophila. The use of 8% caffeine dietary supplementation improved the energetic metabolism via protective effects on CK and AK activities, avoiding the necessity of using anaerobic metabolism. In summary, 8% dietary caffeine can be used to improve branchial energetic homeostasis during aeromonosis caused by A. hydrophila.

PEGylated meloxicam-loaded nanocapsules reverse in vitro damage on caspase activity and do not induce toxicity in cultured human lymphocytes and mice.

Meloxicam is an anti-inflammatory drug that has a potential protective effect in many common diseases. However, this molecule is quickly eliminated from the body due to it short half-life. One way to overcome this problem is to incorporate meloxicam into lipid-core nanocapsules which may increase it anti-inflammatory effects. In view of this, the objective of this work was to evaluate the potential toxicity and safety of these novel nanomaterials both in vitro and in vivo. Here, we evaluated the effects of uncoated meloxicam-loaded nanocapsules (M-NC), uncoated and not loaded with meloxicam or blank (B-NC), PEGylated meloxicam-loaded lipid-core nanocapsules (M-NCPEG), blank PEGylated lipid-core nanocapsules (B-NCPEG) and free meloxicam (M-F) in vitro through the analysis of cell viability, caspase activity assays and gene expression of perforin and granzyme B. Meanwhile, the in vivo safety was assessed using C57BL/6 mice that received nanocapsules for seven days. Thus, no change in cell viability was observed after treatments. Furthermore, M-NC, M-NCPEG and M-F groups reversed the damage caused by H2O2 on caspase-1, 3 and 8 activities. Overall, in vivo results showed a safe profile of these nanocapsules including hematological, biochemical, histological and genotoxicity analysis. In conclusion, we observed that meloxicam nanocapsules present a safe profile to use in future studies with this experimental protocol and partially reverse in vitro damage caused by H2O2.