16S rRNA Analysis of Intestinal Microbiota Alterations in High-Performance Broilers Following Lactobacillus Nebulization

This study aimed to compare the effects of consecutive Lactobacillus nebulization on the pulmonary microbiota diversity of broilers. The two strains were isolated from high-yielding performance broilers, Ligilactobacillus salivarius GHL01 (designated BALF-A), derived from bronchoalveolar lavage fluid (BALF), and Ligilactobacillus salivarius GHL02 (designated Lung-A), isolated from lung tissue. A total of 1,000 one-day-old Arbor Acres (AA) broilers were randomly assigned to three experimental groups, a control group (a total of 200 pieces; 25 replicates, 8 birds per replicate) and two treatment groups receiving aerosolized Lung-A (a total of 320 pieces; 40 replicates, 8 birds per replicate) or BALF-A (a total of 320 pieces; 40 replicates, 8 birds per replicate). A bacterial suspension (the number of bacteria per milliliter is 10^9) was administered via aerosol using a medical nebulizer every three days, with each bird exposed for 20 minutes per session. At 42 days of age, 20 birds were randomly selected from each group and humanely euthanized by cervical dislocation. Cecal content samples were then collected for subsequent 16S rRNA gene sequencing analysis.