2024, Kurhaluk, Natalia, Grudniewska, Joanna, Pękala-Safińska, Agnieszka, Pajdak-Czaus, Joanna, Terech-Majewska, Elżbieta, Platt-Samoraj, Aleksandra, Tkaczenko, Halina

Abstract Introduction This study aimed to evaluate biomarkers of oxidative stress (2-thiobarbituric acid reactive substances, aldehyde and ketone derivatives of oxidatively modified proteins and total antioxidant capacity), the activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase), that of lysosomal enzymes (alanyl aminopeptidase, leucyl aminopeptidase, β-N-acetylglucosaminidase and acid phosphatase) and changes in biochemical parameters (alanine aminotransferase, aspartate aminotransferase, de Ritis ratio, lactate dehydrogenase activity, lactate and pyruvate levels and their ratio) in the liver tissue of fish that were vaccinated against enteric redmouth disease and challenged with its causative agent, the bacterium Yersinia ruckeri. Material and Methods The vaccine was administered orally to trout, some of which were challenged with Y. ruckeri 61 days later. For comparison, unvaccinated and unchallenged trout and unvaccinated and challenged trout were also evaluated. Results In the unvaccinated fish, infection with Y. ruckeri disrupted the pro-oxidant/antioxidant balance, led to a significant increase in lipid peroxidation and oxidative modification of proteins, decreased total antioxidant capacity and significantly increased the activity of lysosomal enzymes. In vaccinated fish, the Y. ruckeri challenge increased the activity of glutathione-related enzymes and decreased lipid peroxidation, anaerobic metabolism and the activity of lysosomal enzymes in fish livers relative to the unvaccinated and challenged group. In contrast, these parameters increased after the Y. ruckeri challenge in unvaccinated trout relative to those in the untreated group. Conclusion Vaccination exerted a protective effect during the Y. ruckeri challenge and had no adverse effect on fish livers.

2025, Tkaczenko, Halina, Kurhaluk, Natalia, Grudniewska, Joanna, Pękala-Safińska, Agnieszka

This study investigated the temporal effects of Yersinia ruckeri vaccination on markers of oxidative stress and antioxidant capacity in the livers of rainbow trout (Oncorhynchus mykiss Walbaum). Oxidative stress was assessed using 2-thiobarbituric acid-reactive substances (TBARS), oxidatively modified protein (OMP) derivatives and total antioxidant capacity (TAC) at three time points: baseline and one and two months post-vaccination. TBARS levels exhibited a transient peak one month after vaccination, suggesting a short-term increase in lipid peroxidation associated with immune activation. In contrast, TAC increased progressively over the experimental period, reaching significantly higher levels in vaccinated fish after two months, which is indicative of enhanced systemic antioxidant defences. OMP derivatives exhibited moderate variability: aldehydic forms peaked in the control group at one month, while ketonic forms were highest at baseline. However, no significant vaccination effect was observed. Correlation analysis revealed a strong negative association between TBARS and TAC (r = −0.58), supporting the role of antioxidant capacity in mitigating oxidative damage. Effect size and variability analyses confirmed substantial time- and treatment-related changes, particularly with regard to TBARS reduction and TAC enhancement in vaccinated fish. Overall, the results demonstrate a biphasic oxidative response to vaccination characterised by an initial oxidative challenge followed by adaptive antioxidant upregulation, highlighting the importance of modulation of redox balance in immune protection and aquaculture health management.

2025, Tkaczenko, Halina, Kurhaluk, Natalia, Grudniewska, Joanna, Pękala-Safińska, Agnieszka, Terech-Majewska, Elżbieta

The liver plays a critical role in maintaining metabolic homeostasis and immune defence in fish, particularly in response to bacterial infections. Autophagy, a conserved cellular process essential for homeostasis and pathogen clearance, has been implicated in host defence mechanisms. However, the role of autophagy in the liver of vaccinated fish following pathogen exposure remains largely unexplored. Yersinia ruckeri, the causative agent of enteric redmouth disease (ERM), poses a significant threat to rainbow trout (Oncorhynchus mykiss Walbaum) aquaculture, primarily affecting the liver, spleen and kidneys. Vaccination is a widely used preventive strategy, but its effect on autophagic activity during infection is not well understood. The aim of this study was to evaluate the autophagic response in the liver of vaccinated rainbow trout following Y. ruckeri infection by assessing the activity of four lysosomal enzymes: alanyl aminopeptidase (AAP), leucyl aminopeptidase (LAP), acid phosphatase (AcP) and β-N-acetylglucosaminidase (NAG). Rainbow trout were divided into experimental groups: unvaccinated control, vaccinated uninfected, unvaccinated infected and vaccinated infected. The fish were orally immunised with a Y. ruckeri vaccine and challenged with a virulent strain of Y. ruckeri. The results showed significant differences in lysosomal enzyme activity between groups, indicating that vaccination modulated the hepatic autophagic response during bacterial infection. AAP and LAP activity peaked in unvaccinated infected fish, whereas vaccinated fish exhibited a blunted enzymatic response, suggesting that vaccination attenuated excessive autophagic activation. Similarly, AcP and NAG activity patterns indicated an infection-induced autophagic response that was partially attenuated in vaccinated fish. These results suggest that vaccination influences autophagy-related enzymatic activity in the liver of rainbow trout, potentially enhancing pathogen clearance while preventing excessive cellular stress. Understanding the interplay between vaccination, infection and autophagy may provide valuable insights to optimise vaccination strategies and improve disease management in aquaculture.