Bacterial community structure in Apis florea larvae analyzed by denaturing gradient gel electrophoresis and 16S rRNA gene sequencing.
This study characterizes the colonization and composition of bacterial flora in dwarf Asian honeybee (Apis florea) larvae and compares bacterial diversity and distribution among different sampling locations. A. florea larvae were collected from 3 locations in Chiang Mai province, Thailand. Bacterial DNA was extracted from each larva using the phenol-chloroform method. Denaturing gradient gel electrophoresis was performed, and the dominant bands were excised from the gels, cloned, and sequenced for bacterial species identification. The result revealed similarities of bacterial community profiles in each individual colony, but differences between colonies from the same and different locations. A. florea larvae harbor bacteria belonging to 2 phyla (Firmicutes and Proteobacteria), 5 classes (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Bacilli, and Clostridia), 6 genera (Clostridium, Gilliamella, Melissococcus, Lactobacillus, Saccharibacter, and Snodgrassella), and an unknown genus from uncultured bacterial species. The classes with the highest abundance of bacteria were Alphaproteobacteria (34%), Bacilli (25%), Betaproteobacteria (11%), Gammaproteobacteria (10%), and Clostridia (8%), respectively. Similarly, uncultured bacterial species were identified (12%). Environmental bacterial species, such as Saccharibacter floricola, were also found. This is the first study in which sequences closely related to Melissococcus plutonius, the causal pathogen responsible for European foulbrood, have been identified in Thai A. florea larvae.
Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand.|Department of Basic Science and Craniofacial Biology, New York University College of Dentistry, New York, 10010, USA.
Prakaimuk
P
Saraithong
Department of Basic Science and Craniofacial Biology, New York University College of Dentistry, New York, 10010, USA.
Yihong
Y
Li
Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand.
Kanokporn
K
Saenphet
Department of Basic Science and Craniofacial Biology, New York University College of Dentistry, New York, 10010, USA.
Zhou
Z
Chen
Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand.|Materials Science Research Center, Faculty of Science, Chiang Mai University, Chiang Mai, 50200, Thailand.
Panuwan
P
Chantawannakul
D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001516:Bees / Q000254:growth & development / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics; D058645:Denaturing Gradient Gel Electrophoresis; D007814:Larva / Q000382:microbiology; D012329:RNA, Bacterial / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013785:Thailand
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S
16S rRNA gene; A. florea; DGGE; gut bacteria community; larvae