Paraquat is a widely used herbicide known to adversely affect honeybee physiology; however, its impact on gut-associated biological processes remains poorly understood. Therefore, intricate effects of paraquat on honeybees highlight the need for a comprehensive understanding of its specific biological consequences. This study aimed to evaluate the gut microbiome composition and metabolomic profiles of honeybees following oral exposure to sublethal (LD25) and toxic (LD50) doses of paraquat, assessed 48 h post-exposure to represent acute exposure. Survival rate and sucrose consumption were monitored to assess toxicity. Honeybee gut samples were analyzed using 16S rRNA gene amplicon sequencing and metabolomic approaches. Both paraquat treatments resulted in increased mortality (p < 0.0001) and reduced sucrose consumption (p = 0.00503). In contrast, gut bacterial community composition remained largely unchanged (p > 0.05). Metabolomic analysis revealed dose-dependent alterations, particularly in metabolites associated with carbohydrate and energy metabolism, including oxaloacetic acid and pyruvic acid. This study provides the first integration of 16S rRNA gene amplicon sequencing and metabolomic analyses to evaluate the effects of paraquat exposure in honeybees. The findings indicate that paraquat-induced stress primarily exhibits as host metabolic reprogramming rather than changes in microbiome composition. These results provide insights into the mechanisms underlying honeybee responses to herbicide stress and may contribute to the development of strategies for honeybee protection.

Melissococcus plutonius, a gram-positive bacterium, causes European Foulbrood (EFB) disease in honeybee larvae, leading to reduced yields in various countries. This study developed a rapid detection method for M. plutonius using colorimetric loop-mediated isothermal amplification with a lateral flow dipstick (cLAMP-LFD), based on the sequences of gyrase subunit B (gyrB) and superoxide dismutase (sodA) genes, giving three primer sets (gyrB1, gyrB2, and sodA). The cLAMP-LFD using gyrB1 was capable of detecting M. plutonius specifically and sensitively under the optimal condition of 63°C for 50 min with no cross-reactivity with other bacteria, and the limit of detection (LOD) was 2 × 104 copies per reaction and 50 CFU of bacterial cells per reaction. In spiked samples, the assay successfully detected M. plutonius in honeybee larvae and adult honeybees, with minimum detectable levels of 11.4 and 80 CFU per reaction, respectively. Our results demonstrate that the gyrB based 1cLAMP-LFD assay is suitable for detecting M. plutonius in apiary settings and in commercial honeybee samples, including those that tested negative by conventional methods. Overall, this cLAMP-LFD assay represents a rapid, sensitive, and specific tool for M. plutonius detection and may facilitate field surveillance, disease control, and management of EFB outbreaks worldwide, thereby supporting improved honey production efficiency.

This study aimed to develop an optimized film-forming gel for topical anti-acne applications by evaluating the antibacterial efficacy of propolis, honey, and royal jelly, individually and in combination with low-dose salicylic acid. The antibacterial activities were assessed against Staphylococcus aureus, Staphylococcus epidermidis, and Cutibacterium acnes using the inhibition zone assay. Film-forming gels were developed by evaluating the effects of type and concentration of polymers and plasticizers. Each formulation was evaluated for visual appearance, pH, viscosity, and drying time, along with the appearance of the corresponding film. The findings noted that propolis (1% w/w) exhibited the strongest antibacterial activity among individual bee products, producing an inhibition zone of 20.0 ± 1.0 mm against S. aureus. The combination of bee products with low-dose salicylic acid (0.1% w/w) markedly enhanced antibacterial efficacy, particularly against C. acnes (inhibition zone 40.8 ± 0.8 mm). Incorporation of this combination into the optimized film-forming gel, containing polyvinyl alcohol, Carbomer® 940, polyethylene glycol 400, glycerin, and water, produced a formulation with balanced pH, suitable viscosity, 31 min drying time, and complete inhibition of S. aureus and S. epidermidis. Therefore, bee product-based film-forming gels, combined with low-dose salicylic acid, exhibited favorable physicochemical properties and showed promise as complementary anti-acne therapies.

Traditional methods for assessing honey storage in beehives predominantly rely on manual visual inspection, which often leads to inconsistencies and inefficiencies. This study presents an automated deep learning approach utilizing the YOLOv11 model to detect, classify, and quantify honey cells within Apis mellifera frames across monthly sampling periods. The model's performance varied depending on image resolution and dataset partitioning. Using the free version of YOLOv11 with high-resolution images (960 × 960 resolution) and a dataset split of 90:5:5 for training, validating, and testing, the model achieved a mean average precision at IoU threshold of 0.5 (mAP@0.5) of 83.4% for uncapped honey cells and 80.5% for capped honey cells. A strong correlation (r = 0.94) was observed between the 90:5:5 and 80:10:10 dataset splits, indicating that increasing the volume of training data enhances classification accuracy. In parallel, the study investigated the relationship between the physical properties of honey and image-based honey storage detection. Of the four tested properties, electrical conductivity (R2 = 0.19) and color (R2 = 0.21) showed weak predictive power for honey storage area estimation, with even weaker associations found for pH and moisture content. The honey storage areas via 90:5:5 and 80:10:10 datasets moderately correlated (r = 0.44-0.46) with increasing electrical conductivity and color. Especially, electrical conductivity exhibited statistically significant correlations with dataset performance across different dataset splits (p < 0.05), suggesting some potential influence of chemical composition on model accuracy. Our findings demonstrate the viability of image-based honey classification as a reliable technique for monitoring beehive productivity. Additionally, the research on image-based honey detection can be a non-invasive solution for improved honey production, beehive productivity, and optimized beekeeping practices.

Tropilaelaps mercedesae ( Mesostigmata : Laelapidae ), an invasive mite native to Southeast Asia, is spreading rapidly due to global bee trade and climate change. Its detection in Eastern Europe raises concerns about its entry into the EU. As a notifiable parasite under EU Animal Health Law, early and reliable detection is essential. We developed a sensitive TaqMan assay targeting the cytochrome C oxidase subunit I (COI) gene, using synthetic double-stranded DNA fragments (gBlocks). This assay enables rapid, specific, and scalable detection of T. mercedesae Clade I. ### Competing Interest Statement The authors have declared no competing interest.

Varroa destructor and Tropilaelaps mercedesae are ectoparasites that pose a significant threat to honeybee populations. It is essential to understand their impact to develop effective management strategies. In this study, we examined the relative abundance of proteins and the expression of immune-related genes in A. mellifera parasitized by the two mites using mass spectrometry analysis of hemolymph and transcriptomics of bleeding honeybees. The study revealed the modulation of genes and proteins associated with immune response and stress management. The apidaecin gene was upregulated in Varroa-infested honeybees and downregulated in Tropilaelaps-infested honeybees, a finding that correlated with MALDI-MS data. Furthermore, variations in the abundance of Hexamerin, Odorant Binding Protein 21, Phenoloxidase and Transferrin were observed in Varroa- and Tropilaelaps-infested honeybees. Using proteomics, potential markers associated with immunity, metabolism and stress responses (e.g. high Glx storage protein, alpha-glucosidase, glyoxylate/hydroxypyruvate reductases) were found to be specific to Varroa and Tropilaelaps infestations. Two Deformed Wing Virus (DWV) proteins exhibit higher abundance in the hemolymph of Tropilaelaps-infested honeybees compared to non-infested and Varroa-infested honeybees, suggesting that Tropilaelaps may weaken honeybee immunity, potentially exacerbating viral infections and colony health decline. This indicates that studying host-mite species interactions could lead to a better understanding of honeybee protection.

Insects often rely on symbiotic bacteria and fungi for various physiological processes, developmental stages, and defenses against parasites and diseases. Despite their significance, the associations between bacterial and fungal symbionts in Apis mellifera are not well studied, particularly in the Philippines. In this study, we collected A. mellifera from two different sites in the Municipality of Bacnotan, La Union, Philippines. A gut microbiome analysis was conducted using next-generation sequencing with the Illumina MiSeq platform. Bacterial and fungal community compositions were assessed using 16S rRNA and ITS gene sequences, respectively. Our findings confirm that adult worker bees of A. mellifera from the two locations possess distinct but comparably proportioned bacterial and fungal microbiomes. Key bacterial symbionts, including Lactobacillus, Bombilactobacillus, Bifidobacterium, Gilliamella, Snodgrassella, and Frischella, were identified. The fungal community was dominated by the yeasts Zygosaccharomyces and Priceomyces. Using the ENZYME nomenclature database and PICRUSt2 software version 2.5.2, a predicted functional enzyme analysis revealed the presence of β-glucosidase, catalase, glucose-6-phosphate dehydrogenase, glutathione transferase, and superoxide dismutase, which are involved in host defense, carbohydrate metabolism, and energy support. Additionally, we identified notable bacterial enzymes, including acetyl-CoA carboxylase and AMPs nucleosidase. Interestingly, the key bee symbionts were observed to have a negative correlation with other microbiota. These results provide a detailed characterization of the gut microbiota associated with A. mellifera in the Philippines and lay a foundation for further metagenomic studies of microbiomes in native or indigenous bee species in the region.

Honey adulteration has long been a nuisance in local and international trade. Sugar syrup addition and false labeling of botanical origin have created a challenge in identifying fraudulent honey supplies and products. Stable carbon isotope ratio analysis (SCIRA) has been widely employed in honey authentication. While it is effective in identifying the addition of C4 plant-derived sugars, it does not provide information related to honey’s botanical source. This research investigated the combination of SCIRA and melissopalynology to provide a more robust assessment of honey integrity and showed that PCA analysis of δ13C together with sugar profiles could further improve the decision involving addition of sugar syrups. A total of 34 beekeeper honey samples were analyzed from 7 provinces of Thailand with a focus on longan honey. Twenty-four samples passed the δ13C criteria, exhibiting δ13C of bulk honey ranging from −28.53 ± 0.19‰ to −22.89 ± 0.08‰ and δ13C of extracted protein ranging from −29.30 ± 0.07‰ to −22.76 ± 0.03‰. Pollen profiling further eliminated honey of questionable and multifloral origins, yielding only eight samples that passed both criteria of being monofloral and not being adulterated with C4-derived sugars. These included six samples of longan honey and two honey samples of other botanical origins, yielding an overall passing rate of 23.5%. Our study showed that by combining SCIRA and melissopalynology, a robust determination of honey integrity could be achieved.

The four honeybee species native to Cambodia—Apis dorsata, Apis cerana, Apis florea, and Apis andreniformis—play a vital role in ecosystem health and agricultural productivity through their pollination activities. Beekeeping in Cambodia has primarily developed around the introduced species Apis mellifera. However, it remains underdeveloped compared to neighboring countries, with wild honey collection continuing to play a significant role. Although native bees are not at immediate risk of extinction, their crucial ecological and socio-economic roles, coupled with the threats to their populations, necessitate the urgent implementation of conservation policies. The beekeeping sector in Cambodia faces numerous challenges that hinder its development. These include a lack of documentation and official data, limited access to productive apiaries, inappropriate pesticide practices, insufficient knowledge about bee health, diseases, and parasites, inadequate organization and representation of beekeepers, the high cost of Cambodian honey, and a lack of consumer awareness and trust in local bee products. This paper provides an overview of the status and distribution of honeybee species, along with the production, value chain, and trade of bee products in Cambodia. It examines the challenges of conserving native honeybees and developing the beekeeping sector while proposing strategies to strengthen bee conservation and support the growth of beekeeping in the country. Key recommendations focus on integrating bee conservation strategies with broader conservation goals, such as reducing deforestation, promoting sustainable agriculture, and regulating the consumption of wild bee brood. Key strategies for the development of beekeeping are documenting and conserving bee floral resources and habitats, reducing the use of bee-toxic pesticides, developing research and training capacities in beekeeping, regulating bee imports, and enhancing quality and authenticity testing facilities. Additionally, building capacity among honey value chain stakeholders in areas such as quality control, authenticity, processing, packaging, and marketing is essential.

This study examined the physicochemical, microbiological, antioxidant, and antibacterial properties of honey derived from stingless bees (Tetragonula pagdeni Schwarz). The honey samples were from different botanical zones in Sa Kaeo province, recognized as a "herb hub" in eastern Thailand. The analysis of 17 stingless bee honey (SBH) samples revelaed an average moisture content of 25.67 + 2.43 %, protein content of 1.99 + 0.11 g/100 g, ash content of 0.77 + 0.42 g/100 g, total soluble solids (TSS) of 77.19 + 1.31 degrees Brix, hydroxymethylfurfural (HMF) level of 0.036 + 0.026 mg/g, pH of 3.9 + 0.19, and free acidity of 127.28 + 52.63. The sugar composition was analyzed, revealing an average glucose content of 19.56 + 7.02 g/100 g, fructose content of 29.52 + 7.19 g/ 100 g, estimated reducing sugars (sum of fructose and glucose; F + G of 49.08 + 13.85), estimated fructose/ glucose ratio (F/G) of 1.60 + 0.37, and estimated glucose/water ratio (G/W) of 0.77 + 0.27. The antioxidant activity was assessed using ABTS and DPPH with average IC50 values of 4.42 + 0.87 and 46.99 + 10.68, respectively. The SBH exhibited an inhibitory effect against Salmonella enterica (10.39 + 3.02 mm), Escherichia coli O157:H7 (9.38 + 1.95 mm), Staphylococcus aureus (11.81 + 5.01 mm), Bacillus cereus DSM4384 (6.90 + 3.55 mm), and Bacillus cereus F4810/72 (10.78 + 2.71 mm) with the agar well diffusion method. Furthermore, the principal components analysis demonstrated that vegetation type influenced the physicochemical and antibacterial properties. Microbiological assessments were conducted to evaluate the quality and food safety of SBH. We analyzed the total bacteria, total yeast and mold, coliforms, E. coli, S. aureus, and presumptive Salmonella spp.. We additionally investigated the culturable bacteria and their enzymatic activities. Bacillus constituted the predominant bacterial isolates from SBH, and numerous isolates exhibited the ability to produce hydrolytic enzymes advantageous for industrial applications.

Stingless bees, a social corbiculate bee member, play a crucial role in providing pollination services. Despite their importance, the structure of their microbiome, particularly the fungal communities, remains poorly understood. This study presents an initial characterization of the fungal community associated with two Thai commercial stingless bee species, Lepidotrigona terminata (Smith) and Tetragonula pagdeni (Schwarz) from Chiang Mai, Thailand. Utilizing ITS amplicon sequencing, we identified distinct fungal microbiomes in these two species. Notably, fungi from the phyla Ascomycota, Basidiomycota, Mucoromycota, Mortierellomycota, and Rozellomycota were present. The most dominant genera, which varied significantly between species, included Candida and Starmerella. Additionally, several key enzymes associated with energy metabolism, structural strength, and host defense reactions, such as adenosine triphosphatase, alcohol dehydrogenase, β-glucosidase, chitinase, and peptidylprolyl isomerase, were predicted. Our findings not only augment the limited knowledge of the fungal microbiome in Thai commercial stingless bees but also provide insights for their sustainable management through understanding their microbiome.

Honey bees provide vital pollination services to agricultural crops and wild plants worldwide. Unfortunately, the misuse and overuse of pesticides in agricultural production have led to an increase in incidents harming honey bees in recent years. Among the commonly utilized bee species in beekeeping are Apis cerana and Apis mellifera, with wild A. cerana populations widely dispersed in forests, contributing substantially to ecosystem balance. Yet, the impact of paraquat, a toxic herbicide, on A. cerana remains largely unexplored. This study aims to address this gap by examining acute exposure endpoints based on mortality represented by median lethal doses (LD50 values) of paraquat, survival rates, and gene expression patterns between the A. cerana and A. mellifera. The findings revealed that A. cerana exhibits greater sensitivity to paraquat compared to A. mellifera. The acute oral LD50 values for A. cerana were 5.85, 1.74, and 1.21 mu g/bee at 24, 48, and 72 h, respectively, whereas the corresponding values for A. mellifera were 104.00, 11.00, and 6.41 mu g/bee. Further, the study demonstrated significant upregulation of the detoxification (antioxidative) enzymes SOD1, CAT, and LLDH-X2 in both A. mellifera and A. cerana following exposure to the lethal dose of paraquat. However, SOD2 expression was notably downregulated in both species, indicating potential mitochondrial damage. These findings suggest that while honey bees initiate activate defense mechanisms against oxidative damage, paraquat exposure may still impair mitochondrial function. Paraquat was found to be moderately toxic to A. mellifera but highly toxic to A. cerana, indicating the importance of screening multiple bee species when assessing the risks of chemical exposure. This research provides a rare comparative analysis of chemical stress effects on morbidity and gene expression in two different honey bee species, establishing a foundational framework for risk assessment and the regulation of herbicide risks to pollinating insects.

Gelatin is commonly used as a gelling agent in gummy candy. Honey and bee products are valuable and rich sources of biologically active substances. In this study, the influence of gelatin and propolis extract on honey gummy jelly (HGJ) properties was investigated. Honey (28–32%), xylitol (13–17%), and gelatin (6–10%) were utilized to develop HGJ products by mixture design methodology. Subsequently, the optimized formulation of HGJ was fortified with 1% and 2% propolis extract to enhance its phytochemicals and antimicrobial activities. The variation in the ingredients significantly affected the physicochemical, textural, and sensory properties of the HGJ. The optimized HGJ formulation consisted of honey (32%), xylitol (14%), and gelatin (7%) and exhibited 13.35 × 103 g.force of hardness, −0.56 × 103 g.sec of adhesiveness, 11.96 × 103 N.mm of gumminess, 0.58 of resilience, and a moderate acceptance score (6.7–7.5). The fortification of HGJ with propolis extract significantly increased its phytochemical properties. Furthermore, the incorporation of propolis extract (2%) into the HGJ was able to significantly inhibit the growth of Gram-positive (Streptococcus mutans and Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria. The mixture of gelatin, xylitol, honey, and propolis extract can be utilized to develop a healthy gummy product with acceptable physicochemical, textural, and sensory qualities.

This research paper aims to explore a network governance in sustainable apiculture management and identify its challenges to government and non-governmental policy makers. Qualitative data was collected through participant observation during field visits and Focus Group Discussions (FGDs) with regional apiculture management. The research results show that each country in the Mekong subregion has different network governances in supporting a sustainable apiculture management. Some countries have a collaborative network between the government, the civil society, entrepreneurs, and scholars. In contrary, some countries lack a policy framework from the government. This difference becomes challenges for the six governments of the Mekong subregion countries to commonly support the sustainable apiculture management in the subregion. The identified challenges in the paper are: (1) a capacity gap between the six countries of the region; (2) no common policies and regulations that would facilitate local beekeepers’ access to wider regional bee products; (3) no support for inter-regional transport of bees and honey for academic purposes; (4) inadequate native bee species preservation and pesticides use; (5) local beekeepers’ limited access to a regional bee diseases and pesticides use database; and, (6) a lack of regional standards for general honeybee product support.

Stingless bees play a crucial role in the environment and agriculture as they are effective pollinators. Furthermore, they can produce various products that can be exploited economically, such as propolis and honey. Despite their economic value, the knowledge of microbial community of stingless bees, and their roles on the bees' health, especially in Thailand, are in its infancy. This study aimed to investigate the composition and the functions of bacterial community associated with Tetragonula pagdeni stingless bees using culture-independent and culture-dependent approaches with emphasis on lactic acid bacteria. The culture-independent results showed that the dominant bacterial phyla were Firmicutes, Proteobacteria and Actinobacteria. The most abundant families were Lactobacillaceae and Halomonadaceae. Functional prediction indicated that the prevalent functions of bacterial communities were chemoheterotrophy and fermentation. In addition, the bacterial community might be able to biosynthesize amino acid and antimicrobial compounds. Further isolation and characterization resulted in isolates that belonged to the dominant taxa of the community and possessed potentially beneficial metabolic activity. This suggested that they are parts of the nutrient acquisition and host defense bacterial functional groups in Thai commercial stingless bees.

Widespread parasites, along with emerging threats, globalization, and climate change, have greatly affected honey bees' health, leading to colony losses worldwide. In this study, we investigated the detection of biotic stressors (i.e., viruses, microsporidian, bacteria, and fungi) in Apis cerana by surveying the colonies across different regions of Thailand (Chiang Mai in the north, Nong Khai and Khon Kaen in the northeast, and Chumphon and Surat Thani in the south, in addition to the Samui and Pha-ngan islands). In this study, we detected ABPV, BQCV, LSV, and Nosema ceranae in A. cerana samples through RT-PCR. ABPV was only detected from the samples of Chiang Mai, whereas we found BQCV only in those from Chumphon. LSV was detected only in the samples from the Samui and Pha-ngan islands, where historically no managed bees are known. Nosema ceranae was found in all of the regions except for Nong Khai and Khon Kaen in northeastern Thailand. Paenibacillus larvae and Ascosphaera apis were not detected in any of the A. cerana samples in this survey. The phylogenetic tree analysis of the pathogens provided insights into the pathogens' movements and their distribution ranges across different landscapes, indicating the flow of pathogens among the honey bees. Here, we describe the presence of emerging pathogens in the Asian honey bee as a valuable step in our understanding of these pathogens in terms of the decline in eastern honey bee populations.

Widespread native honey bee species in South and East Asia (Apis cerana, Apis dorsata and Apis florea) and the imported western honey bee (Apis mellifera) share habitats and potentially also share pathogens. Chief among the threats facing A. mellifera in Europe and North America is deformed wing virus (DWV), including its two prin-cipal genotypes: A and B (DWV-A and DWV-B respectively). Though DWV-A has been recorded in Asia's native Apis species, it is not known if DWV-B, or both DWV-A and DWV-B, are currently widespread in Asia and, if so, whether viral transmission is primarily intraspecific or interspecific. This study aims to fill these knowledge gaps by (i) determining the DWV genotype in four co-occurring Apis host species using qPCR and (ii) inferring viral transmission between them using nucleotide sequences of DWV from Apis host species collected at three inde-pendent localities in Northern Thailand. We found DWV-A and-B in all four Apis species, the exotic A. mellifera and the native A. cerana, A. dorsata and A. florea. That DWV-A sequences were identical across Apis species at the same locality, with a similar pattern for DWV-B sequences, suggests that DWV's epidemiology is largely driven by ongoing interspecific transmission (spillover) of DWV across co-occurring native and exotic Apis species. Both genotypes of DWV represent a serious threat to Asia's exotic and native honey bee species.

Tropilaelaps mercedesae, one of the most devastating parasitic mites of honey bee Apis mellifera hosts, is a major threat to honey products by causing severe damage to honey bee colonies. Here, we recorded injury numbers caused by T. mercedesae to different body parts of the larval, pupal, and crippled adult stages of honey bee A. mellifera. We evaluated the relationship between infestation rate and injury numbers per bee for both larvae and pupae. We also noted the total bee numbers per beehive and examined the relationship between the infestation rate and population size. T. mercedesae infested all developmental stages of honey bees, with the highest injury numbers in the abdomens of bee pupae and the antennas of crippled adult bees. Although larvae received more injury numbers than pupae, both infestation rate and injury numbers decreased as the larval stage progressed to the pupal stage. The infestation rate increased as the population size per beehive decreased. This study provided new perspectives to the understanding of changes in the effects of T. mercedesae infestations on different developmental stages of honey bees. It also showed useful baseline information for screening honey bee stock that might have high defensive behaviors against mite infestation.

Edible insects have become increasingly popular in Thailand as a nutritious and appealing alternative food source. As the edible insect industry in the country expands rapidly, efforts are being made to transform it into an economically viable sector with substantial commercial potential. Some of the most consumed and sold edible insects in Thailand include locusts, palm weevils, silkworm pupae, bamboo caterpillars, crickets, red ants, and giant water bugs. With its strong growth, Thailand has the potential to emerge as a global leader in the production and promotion of edible insect products. Edible insects are an excellent source of protein, fat, vitamins, and minerals. In particular, crickets and grasshoppers are protein-rich, with the average protein content of edible insects ranging from 35 to 60 g/100 g of dry weight or 10 to 25 g/100 g of fresh weight. This surpasses the protein content of many plant-based sources. However, the hard exoskeleton of insects, which is high in chitin, can make them difficult to digest. In addition to their nutritional value, edible insects contain biologically active compounds that offer various health benefits. These include antibacterial, anti-inflammatory, anti-collagenase, elastase-inhibitory, α-glucosidase-inhibitory, pancreatic lipase-inhibitory, antidiabetic/insulin-like/insulin-like peptide (ApILP), antidiabetic, anti-aging, and immune-enhancing properties. The Thai food industry can process and utilize edible insects in diverse ways, such as low-temperature processing, including refrigeration and freezing, traditional processing techniques, and incorporating insects into products, such as flour, protein, oil, and canned food. This review offers a comprehensive overview of the status, functional properties, processing, and utilization of edible insects in Thailand, and it serves as a valuable resource for those interested in edible insects and provides guidance for their application in various fields.

This study aimed to develop a film-forming gel containing honey aromatic water (HW) and royal jelly (RJ) for cosmetic applications as a facial peel-off mask. HW, which is industrial waste from the water-reduction process of honey, was sterilized by autoclaving and filtration through a 0.22 µm membrane. The film-forming gels were developed using various types of film-forming polymers, including polyvinyl alcohol (PVA 117), carboxymethyl cellulose (CMC), and hydroxyethyl cellulose (HEC). The gel formulations were characterized in terms of their external appearance, viscosity, pH, and drying time, whereas the films generated were characterized by a texture analyzer, microscopic investigation, Fourier transform infrared, and an X-ray diffractometer. The findings highlighted that HW has short storage shelf life due to microbial contamination. Sterilizations were required before further product development. The film-forming gel was created by using the combination of PVA 117, CMC, and HEC. HW and RJ were successfully incorporated into the film-forming gel. However, HW resulted in a decrease in the gel viscosity and mechanical properties of its film. Interestingly, the drying time was dramatically decreased, which would be more desirable for its use as a peel-off mask. Furthermore, incorporation of royal jelly enhanced the viscosity of the gels as well as improved the mechanical properties of the film. No effect on the chemical and crystal structure of the films was detected after the incorporation. Therefore, the film-forming gels containing HW and RJ, possessing aesthetic attributes that extended to both the gels themselves and the resultant films, were suitable for use as a peel-off mask.