The silkworm (Bombyx mori) is an economically important insect and serves as a model organism for Lepidoptera. To investigate the effects of the intestinal microbial population on the growth and development of larvae fed an artificial diet (AD) during the young stages, we analyzed the characteristics of the intestinal microbial population using 16S rRNA gene sequencing technology. Our results revealed that the intestinal flora of the AD group tended to be simple by the 3rd‐instar, which Lactobacillus accounting for 14.85% and leading to a decreased pH in the intestinal fluid. In contrast, the intestinal flora of silkworms in the mulberry leaf (ML) group showed continuous growth of diversity, with Proteobacteria accounting for 37.10%, Firmicutes accounting for 21.44%, and Actinobacteria accounting for 17.36%. Additionally, we detected the activity of intestinal digestive enzymes at different instars and found that the activity of digestive enzymes in the AD group increased by larval instar. Protease activity in the AD group was lower during the 1st‐ to 3rd‐instars compared to the ML group, while α‐amylase and lipase activities were significantly higher in the AD group during the 2nd‐ and 3rd‐instar compared to the ML group. Furthermore, our experimental results indicated that changes in the intestinal population decreased the pH and affected the activity of proteases, which might contribute to the slower growth and development of larvae in the AD group. In summary, this study provides a reference for investigating the relationship between artificial diet and intestinal flora balance.

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Glyphosate is a widely used herbicide and crop desiccant.However, whether its extensive use has any effect on the species diversity of nontarget organisms is still unclear. In this study, we used the silkworm, Bombyx mori, as the

The Duox-ROS defense system plays an important role in insect intestinal immunity. To investigate the role of intestinal microbiota in Duox-ROS regulation herein, 16S rRNA sequencing technology was utilized to compare the characteristics of bacterial populations in the midgut of silkworm after different time-periods of treatment with three feeding methods: 1–4 instars artificial diet (AD), 1–4 instars mulberry leaf (ML) and 1–3 instars artificial diet + 4 instar mulberry leaf (TM). The results revealed simple intestinal microbiota in the AD group whilst microbiota were abundant and variable in the ML and TM silkworms. By analyzing the relationship among intestinal pH, reactive oxygen species (ROS) content and microorganism composition, it was identified that an acidic intestinal environment inhibited the growth of intestinal microbiota of silkworms, observed concurrently with low ROS content and a high activity of antioxidant enzymes (SOD, TPX, CAT). Gene expression associated with the Duox-ROS defense system was detected using RT-qPCR and identified to be low in the AD group and significantly higher in the TM group of silkworms. This study provides a new reference for the future improvement of the artificial diet feeding of silkworm and a systematic indicator for the further study of the relationship between changes in the intestinal environment and intestinal microbiota balance caused by dietary alterations.