Understanding the honeybee gut bacteria is an essential aspect as honeybees are the primary pollinators of many crops. In this study, the honeybee-associated gut bacteria were investigated by targeting the V3-V4 region of 16S rRNA genes using the Illumina MiSeq. The adult worker was captured in an urban area in a dense settlement. In total, 83,018 reads were obtained, revealing six phyla from 749 bacterial operational taxonomic units (OTUs). The gut was dominated by Proteobacteria (58% of the total reads, including Enterobacteriaceae 28.2%, Erwinia 6.43%, and Klebsiella 4.90%), Firmicutes (29% of the total reads, including Lactococcus garvieae 13.45%, Lactobacillus spp. 8.19%, and Enterococcus spp. 4.47%), and Actinobacteria (8% of the total reads, including Bifidobacterium spp. 7.96%). Many of these bacteria belong to the group of lactic acid bacteria (LAB), which was claimed to be composed of beneficial bacteria involved in maintaining a healthy host. The honeybee was identified as Apis nigrocincta based on an identity BLAST search of its COI region. This study is the first report on the gut microbial community structure and composition of A. nigrocincta from Indonesia.
Introduction: Apis nigrocincta is a honeybee endemic to Mindanao island (the Philippines), Sangihe island (North Sulawesi, Indonesia) and Sulawesi mainland (Indonesia). The genus Apis is well known to have symbiont in their guts, which helps balance the microbiome in the gut and host health. Objective: The objective of this study was to determine whether the bacteria isolated from the gut of honeybee Apis nigrocincta produce metabolites with potential growth inhibition against Staphylococcus aureus and Escerichia coli, the bacteria which are important pathogens in humans and animals. Methods: Bacteria isolated from honeybee gut were cultured in MRSA and several isolates were purified for testing. The antibacterial activity test method used in this study was well diffusion agar. Pure isolates were grown on NB. The treatments given were heating and also neutralizing the supernatant from each isolate. Results: Five bacterial isolates were successfully isolated from honeybee gut and purified. The five isolates showed antibacterial activity against pathogenic bacterial strain indicators. The results of molecular identification showed that four of these isolates were Bacillus cereus and the other one was Staphylococcus arlettae. Neutralized supernatant showed strong activity on both indicator strains. The five isolates showed higher inhibition activity against S. aureus compared to E. coli. Conclusion: The finding of this research concluded that two bacterial strains, B. cereus and S. arlettae isolated from A. nigrocincta gut can be investigated further as agents which produce bioactive compounds that have potential as an antibacterial.
Asian cavity-nesting honey bee Apis nigrocincta, a native bee species from Sulawesi and the Philippines, plays a vital role in pollinating flowering plants in local ecosystem and agriculture. In this study, we assessed the intraspecific genetic variation of A. nigrocincta using the sequence of cytochrome c oxidase subunit I (COI). Molecular phylogenetic analysis showed that there were three main clades in A. nigrocincta specimens from Sulawesi based on their respective locations (North, Central, and South Sulawesi). Genetic distance analysis using the Kimura 2-parameter (K2P) model showed that the intraspecific genetic distance in Sulawesi specimens ranged from 0.000 to 0.055. There are 26 nucleotide polymorphic sites within Sulawesi A. nigrocincta. The variation was dominated by transition T ↔ C. The molecular identification result was supported by morphological identification. The results of the two methods agree that the specimen under study was A. nigrocincta. The result of genetic distance calculation indicated that although the tested specimens were derived from remote locations, the genetic variation was still within the range of intraspecific variation.
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