The effect of higher packing density and increased duration of transport on the survival and key metabolic enzymes of Labeo rohita fry was investigated. L. rohita fry (length 40 +/- A 5 mm, weight 0.60 +/- A 0.13 g) were packed in two different densities 40 and 80 g/l and sampled at 0, 12, 24, and 36 h after packing. Results showed that packing density and length of confinement severely affected the survival of the fry. The whole-body glucose level and the activities of the enzymes, lactate dehydrogenase (LDH), malate dehydrogenase (MDH), glucose-6-phosphatase (G6Pase), fructose-1, 6-bisphosphatase (FBPase), aspartate amino transferase (AST), alanine amino transferase (ALT), and adenosine triphosphatase (ATPase) assayed from the fish whole-body significantly (P < 0.05) increased due to increase in the length of the confinement. However, acetylcholine esterase (AchE) activity decreased significantly (P < 0.05) with increase in the length of confinement. Similarly, higher packing density also significantly (P < 0.05) increased the glucose level and activities of all these enzymes (except AchE). The results revealed that both higher packing density and increased transportation duration mobilize protein resources for glucose production via gluconeogenesis and subsequently activate the glycolysis pathway for energy. The rise in the ATPase activity indicates disruption of the osmoregulatory function and the role of this enzyme in ameliorating it. Overall results suggest that normally practiced packing density of 40 g/l is optimum up to 24-h duration for seed transportation