Excess applications of chemical fertilizer give adverse effects on regional ecosystems and environment. We studied the effects of mulberry leaves grown at different fertilizer application rates, including long-term unfertilized cultivation, on the susceptibility of silkworm larvae to a food dye, phloxine (C.I. Acid Red 92), which is also known as a carcinogenic substance. Silkworm larvae raised with unfertilized mulberry leaves showed about 10 times stronger resistance to phloxine than those raised with fertilized leaves, living for longer times after fed on leaves treated with 4% phloxine solution. Median lethal time of silkworm larvae after phloxine application significantly differed among different fertilizer application rates in the following order: no nitrogen (N), no phosphate (P) and no potassium (K) > no N but with P and K < with N, P and K. With respect to free amino acids of mulberry leaves, the concentrations of Asp, Gln, Thr, His, Pro, Gly and Ala were positively correlated to the nitrogen application rate.
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