Gut bacteria are essential to honey bee (Apis mellifera) health, supporting digestion, immunity, and resilience against stressors. Probiotic-based strategies have been proposed to enhance core gut symbionts, yet the underlying mechanisms and influences of environmental context on these effects are not fully understood. This study examined the influence of gut-derived bacterial supplementation on the honey bee gut microbiome, proteome, and metabolome across three conditions: (i) controlled cages, (ii) semi-controlled cages allowing contact with nestmates, and (iii) field conditions. Treatment groups received a bacterial supplement containing Lactobacillus helsingborgensis, Lactobacillus apis, Bifidobacterium choladohabitans, and Bifidobacterium polysaccharolyticum, while control groups received only a sucrose solution. Gut samples from 10-day-old bees were analyzed. Supplemented bees showed strong gut colonization by Bifidobacterium and Lactobacillus, especially under controlled and semi-controlled conditions. In the field, L. helsingborgensis remained significantly enriched in the treatment group, demonstrating short-term ecological resilience in the natural hive environment. Proteomic changes in supplemented bees included an increased abundance of major royal jelly protein precursors and mitochondrial-associated proteins, together with reduced abundance of several ribosomal and translation-related proteins involved in peptide biosynthesis and cellular protein quality control. Metabolomic analysis revealed reproducible shifts across all three conditions. Treatment groups showed higher concentrations of microbial fermentation products (acetate, succinate) and potential neuromodulatory metabolites (ornithine, γ-aminobutyrate), while sucrose, N-acetylglucosamine, and uridine were constantly lower compared to controls. These findings highlight reproducible, context-dependent effects of bacterial supplementation on honey bee gut physiology and provide a framework for interpreting microbiome-based interventions in pollinator health.IMPORTANCEHoney bees are essential pollinators whose health is influenced by their gut microbiome. Probiotic applications aimed at improving gut health have been proposed, yet outcomes remain inconsistent and vary across settings. Results from laboratory experiments often differ from those observed under field conditions, making it difficult to understand the complex dynamics of eusocial insect colonies. Here, we evaluate honey bee gut-derived bacterial supplementation across controlled, semi-controlled, and field settings using bacterial profiling, proteomic, and metabolomic analyses. We demonstrate that bacterial-supplemented groups consistently reshape gut community composition and modulate host physiological processes, but in a context-dependent manner. These results provide a unified understanding of how microbial interventions function at colony and individual levels, guiding the rational design of probiotic strategies to support honey bee health under realistic conditions.
The Western honey bee plays a pivotal role in global food security as the primary commercial pollinator. The microsporidian pathogens Nosema apis and Nosema ceranae infect the bee midgut, causing nosemosis, a debilitating infectious disease that results in considerable economic losses in apiculture. Traditionally, Nosema spp. infection is diagnosed by microscopic detection and quantification of spores. However, only molecular diagnostics allow differentiation between N. apis and N. ceranae. Loop-mediated isothermal amplification (LAMP) is a rapid, highly specific, and sensitive DNA detection method. The present study aimed to develop a LAMP protocol for N. apis based on the species-specific single copy polar tube protein 2 (ptp2) gene, and to analyze and compare its diagnostic performance with the previously developed polar tube protein 3 (ptp3) gene-based LAMP protocol for N. ceranae. The ptp2- and ptp3-LAMP assays specifically identified N. apis and N. ceranae, respectively. Their analytical sensitivity was tested using serial dilutions of plasmid and genomic DNA, demonstrating that ptp2- and ptp3-LAMP consistently detected down to 103ptp2 and 104ptp3-gene copies, respectively. Amplification was verified by agarose gel electrophoresis (conventional format), and by a change from pink to yellow color after addition of a suitable dye (colorimetric format). The ptp2- and ptp3-LAMP assays and a reference duplex PCR were applied to a panel of field samples (n = 55) from a region endemic for both Nosema spp. Conventional and colorimetric ptp2-LAMP showed an almost perfect test agreement (kappa value > 0.81) compared with duplex PCR. Conventional and colorimetric ptp3-LAMP assays showed a substantial (kappa value > 0.60) and almost perfect test agreement (kappa value > 0.81), respectively. The ptp2- and ptp3-LAMP assays provide excellent performance, ease of implementation, cost savings, and rapid execution, making them ideal choices for molecular detection and differentiation of N. apis and N. ceranae.
Background: Indonesia is currently facing a public health concern, possessing the fifth-highest prevalence of individuals with Type 2 Diabetes Mellitus (T2DM) worldwide. Type 2 diabetes mellitus (T2DM) is significantly linked to problems with oral health that can greatly lower patients' Oral Health-Related Quality of Life (OHRQoL). Nonetheless, a thorough synthesis of the evidence in the Indonesian context is absent. Objective: This study aim to comprehensively synthesize the Oral Health-Related Quality of Life of patients with Type 2 Diabetes Mellitus in Indonesia. Method: A systematic literature review was performed by querying PubMed, Scopus, and Cochrane Library for papers published from January 2004 to December 2025. The search encompassed observational and interventional studies conducted in Indonesia that evaluated OHRQoL in adult T2DM patients utilizing validated measures. A narrative synthesis of the findings was conducted due to methodological heterogeneity. Results: The review of 6 studies revealed a consistent trend of reduced OHRQoL among Indonesian T2DM patients, despite variations in overall scores among studies. The OHRQoL domains that were most often and most seriously affected were physical pain and functional restrictions, like trouble chewing. Periodontitis, tooth mobility, and xerostomia were some of the most important oral manifestations that caused this effect. Glycemic control, diabetes duration, and lower educational attainment were important factors contributing to reduced OHRQoL. Conclusion: T2DM substantially detrimentally affects the OHRQoL of patients in Indonesia, chiefly via functional and physical dimensions. These results highlight the pressing necessity of including oral healthcare into national chronic disease management plans to enhance patient-centered outcomes.
