Diversity of bacteria in the honey stomach of Apis cerana and Apis mellifera during the rape blooming period
Author of the article:WANG Miao1, 2** XU Hong2 YIN Ge-Fen3 ZHAO Wen-Zheng2 HE Shao-Yu2***
Author's Workplace：1. Plant Protect College, Yunnan Agricultural University, Kunming 650201, China; 2. Eastern Bee Research Institute, YunnanAgricultural University, Kunming 650201, China; 3. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China
Key Words：Apis cerana, Apis mellifera, honey stomach, 16S rDNA, bacterial diversity
Abstract：[Objectives] To analyze the diversity, origin and differences in the cultured bacterial community present in the honey stomach of Apis cerana and Apis mellifera. [Methods] Bacteria were isolated and cultivated to obtain pure isolates in the honey stomach of A.cerana and A. mellifera. The isolates were compared and a phylogeny constructed analysis based on 16S rDNA sequences amplification and sequencing. [Results] Bacillus amyloliquefaciens was the dominant bacteria comprising 69.81% to 95.97% of the total found in each bee species respectively, significantly higher than other bacteria, which comprised just 4.03% to 30.19%. P-values for these differences were 0.007 to 0.009, respectively. A total of 275 bacterial isolates were selected and identified from different phenotypes. All bacterial 16S rRNA sequences from the different samples showed nine haplotypes and >99% similarity to the phylotype in GenBank. 16 isolates were submitted to GenBank and obtained accession numbers from kf710012 to kf710033. The honey stomach of A. cerana contained five bacterial species from two genera, in comparison, the honey stomach of A. mellifera contained seven species from five genera and nectar contained two species from two genera. Phylogenetic analysis indicated that clusters Ⅰand Ⅱ were the same species as the previously described EF565944、EF565939 and EF565936. Cluster Ⅴ(Lactobacillus) was not found in this study. Clusters Ⅲ,Ⅳ, Ⅵ and Ⅶ were also not found in the reference strains. This is the first time that bacteria in the honey stomach of A. cerana and A. mellifera have been described and submitted to GenBank. [Conclusion] Except for the common dominant bacteria B. amyloliquefaciens，bacterial diversity in the honey stomach is obviously different between A. cerana and A. mellifera. Comparison with previous, overseas studies of the honey stomach bacterial community suggests that there are geographic differences in bacterial community composition. B. amyloliquefaciens can be used as a food additive that enhances disease resistance in honeybees.