Toxicity of nitenpyram to silkworm (Bombyx mori L.) and its potential mechanisms

Silkworm (Bombyx mori L.), as an economic insect, occupies a certain position in the development of China's economy. The neonicotinoid insecticide nitenpyram is commonly used in farmland to control planthoppers and aphids. In China, mulberry orchards are often planted adjacent to fields or commercial crops, and mist drifts occur during application, which may affect the production safety of Bombyx mori. In this study, a risk assessment of nitenpyram was carried out, and the results showed that there were risks in spraying nitenpyram around the periphery and subperipheries of mulberry fields. However, few studies have reported the mechanism underlying nitenpyram's toxic effect on silkworms. Here, we validated 25 differentially expressed (DE) miRNAs in the nitenpyram treatment group of silkworms, and the significantly enriched mTOR signaling pathway, oxidative phosphorylation and FoxO signaling pathway were verified. Among them, bmo-miR-2766-5P was up-regulated by 2.122-fold, and the expression of its regulated target gene 101,741,287 was up-regulated. After the injection of bmo-miR-2766-5P inhibitor, the Log₂FC value of 101,741,287 was changed from 1.26 to -2.19. Bmo-miR-3326, bmo-miR-3378-5P and bmo-miR-2761-3P were down-regulated by 2.386-fold, 1.158-fold and 2.359-fold, respectively. After injecting miRNA mimics into silkworms, the Log₂FC values of the target genes 100,302,609, 101,740,730 and 101,746,319 were changed from 1.24 to -11.94, -1.12 changed to 2.84 and 1.93 changed to -0.37, respectively. In addition, nitenpyram induced oxidative damage in silkworms, and the degree of DNA damage increased with the increase of concentration and time. Meanwhile Imd was significantly up-regulated in IMD-related pathways (38.7-fold, p < 0.01). The results indicated that nitenpyram could affect the growth and development process of silkworms, and these DE-miRNAs may have an important impact on the stress response of silkworms to nitenpyram.