Increasing stocking density causes inhibition of metabolic–antioxidant enzymes and elevates mRNA levels of heat shock protein 70 in rainbow trout

The objective of this study was to determine the effects of stocking density on the activity of glucose 6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD) and glutathione reductase (GR) enzymes in liver, muscle, gill and kidney tissues of rainbow trout. Fish were reared at different stocking densities (15 kg/m³, 20 kg/m³, 25 kg/m³ and 30 kg/m³). After adaptation period of 30 days, the experiment was carried out for two months. Stocking density of the control group was 15 kg/m³. Increasing stocking density caused inhibition in the activities of the enzymes except for kidney G6PD and 6PGD. Activity of both pentose phosphate pathway enzymes unexpectedly increased only in kidney whereas inhibition was observed in other tissues. Since the most powerful and gradual attenuation was observed in muscle tissue for all enzymes, we performed quantitative Real Time PCR to examine the expression of heat shock protein (Hsp70) gene in muscle in order to understand whether the decrease in enzyme activities is associated with stress. The mRNA expression data showed that Hsp70 expression levels significantly elevated at 25 kg/m³ and 30 kg/m³ stocking densities. Overall results indicate that increasing stocking density blocks the activity of metabolic and antioxidant enzymes and causes considerable stress in rainbow trout. The most susceptible tissue to increasing stocking density was observed to be the muscle.