Ultra-morphology of the scale as an indicator of the stress of Acid Black-1 (AB-1, CI: 20470) and zinc (Zn)

High contents of azo dyes and heavy metals enter surface waters with the wastewater from dying and dye-manufacturing industries and pose serious threat to fish. In the present study, changes in the ultra-morphological features of the scale have been evaluated as indicators of the stress of lethal and sublethal concentrations of an azo dye Acid Black-1 (AB-1, CI 20470), zinc (Zn), and their mixture AB-1 + Zn to Labeo rohita. Fish were exposed for 96 h to lethal concentration (LC) causing 0–70% mortality, i.e., LC₀, LC₂₀, LC₅₀, and LC₇₀ of AB-1 (4, 6, 8, and 10 mg/L respectively) and Zn (25, 50, 55, and 60 mg/L respectively) and LC₀, LC₅₀, and LC₇₀ of AB-1 + Zn (2 + 15, 2 + 20, and 2 + 25 mg/L respectively). Subchronic exposures of 150 days were given to 1/12, 1/6, and 1/3 of 96 h LC₅₀ values of AB-1, Zn, and AB-1 + Zn. After each exposure, the fish were kept for a recovery period of 90 days. Breakage of circuli, erosion and breakage of lepidonts, uprooting of tubercles, and disappearance of intercircular teeth were observed in all the fish, after 96 h exposure to AB-1, Zn, and AB-1 + Zn. However, damage to focus and holes were common on the scales of Zn-exposed fish. The mixture AB-1 + Zn was more toxic than either of the two as loss of circuli, lepidonts, and intercircular teeth, and sloughing of surface were observed in the scales after 96 h exposure to 2 + 25 mg/L. Damage at this concentration was more than the damage at 8/10 mg/L AB-1 and 55/60 mg/L Zn. After 150 days of exposure, damage due to sublethal concentrations was more than the damage due to all the concentrations of 96 h exposure. Irregular and sloughed circuli were common at 2 + 1.79 and 2 + 3.59 mg/L AB-1 + Zn. Cracks all over the surface (2 + 7.18 mg/L), calcium projections (2 + 3.59 and 2 + 7.18 mg/L), and holes (2 + 7.18 mg/L) hint towards synergistic toxicity of the mixture. It seems that the present dye and metal formed complexes with collagen and osteoblastic cells of the scale that caused an increase in damage during the post-exposure period. Scales of 2 + 7.18 mg/L AB-1 + Zn exposed fish were completely devoid of the normal architectural pattern on the 90th day of the recovery period. Changes in the ultra-morphology of scales at LC₀ (0% mortality) and sublethal concentrations show that these are early indicators of the stress of minute quantities of dyes and metals in water. This is a first report on the cumulative toxicity of the two most abundant components of textile industry effluents.