# The Inclusion of Hermetia Illucens Larvae Meal in the Diet of Laying Hens (Hy‐Line Brown) Affects the Caecal Bacterial Composition and Diversity

## TL;DR

Replacing soybean meal with Hermetia illucens larvae meal in laying hens' diets changes gut bacteria, with 50% replacement showing the most benefits.

## Contribution

This study shows that 50% soybean meal replacement with H. illucens larvae meal positively alters the gut microbiota of laying hens.

## Key findings

- Beta diversity analysis showed distinct microbial profiles among the treatment groups.
- The 50% replacement group had enriched beneficial genera and reduced harmful taxa like Enterococcus.

## Abstract

The poultry industry is increasingly looking for sustainable feed ingredients which support animal productivity. Insects, such as the larvae of Hermetia illucens, represent a promising alternative protein source that has a positive effect on gut health.

The aim of this study is to evaluate the effects of a partial replacement of soybean meal with H. illucens larvae meal on the caecal microbiota of laying hens.

A total of 162 hens were divided equally into three treatment groups: a control group (C) with a diet containing corn–soybean meal and two treatment groups (HI25, HI50) in which 25% and 50% of the soybean meal protein was replaced by H. illucens larvae meal protein. At 40 weeks of age, 30 animals (10 per group) were slaughtered and the bacterial community of the caecal content was analysed by high‐throughput sequencing using the V4–V5 region of the 16S rRNA gene. The DNA was extracted using the PowerSoil DNA Kit, the library preparation was performed using the NEBNext Fast DNA Library Prep Set kit and sequencing was performed using the Ion Torrent PGM. The bacterial diversity was assessed by alpha and beta diversity indices, and the differential abundance of taxa was determined using LEfSe analysis.

Firmicutes and Bacteroidetes were the dominant phylum in all groups. Alpha diversity indices showed no significant differences between diets, however, beta diversity measures showed statistical dissimilarities between the three studied groups. Several beneficial genera, including Alistipes, Christensenellaceae R‐7 group, Parabacteroides, Butyricimonas and Parasutterella, were enriched in the HI50 group, while Lactobacillus, Bifidobacterium, Blautia and the opportunistic pathogen Enterococcus were reduced.

The consumption of H. illucens larvae meal showed beneficial effect on the microbiota of the caecum of laying hens, with 50% replacement showing the strongest positive effects, suggesting that this is the most effective amount under the given conditions. Further research should explore microbial functions and long‐term impacts, and validate the optimal levels of insect meal inclusion.

This study evaluated partial replacement of soybean meal with Hermetia illucens larvae meal in laying hens’ diets. While alpha diversity was unaffected, beta diversity revealed distinct microbial profiles among treatments. A 50% replacement enriched beneficial genera (Alistipes, Parabacteroides, Butyricimonas) and reduced potentially harmful taxa (Enterococcus), indicating positive modulation of caecal microbiota.