Aktivitas Antiinflammasi Kombinasi Ekstrak Propolis dan Madu Lebah Heterotrigona itama pada Mencit (Mus musculus) yang diinduksi Karagenan

Previous studies have reported artemisinin resistance. Drugs for malaria treatment have been developed from plants. Citrus aurantifolia was traditionally used to treat ailments. This study aimed to evaluate an antimalarial effect from Citrus aurantifolia, honey and a combination of Citrus aurantifolia and honey  against Plasmodium berghei and analyze phytochemicals from the formula.  Thirty mice were randomized into five groups. A four-day suppressive test was performed to evaluate the antimalarial activity. Data were analyzed using one- way ANOVA followed by Tukey’s post hoc-test to compare between and within  groups. Test results which show a p-value of <0.05 were considered statistically  significant. The suppressive model showed a percentage parasite suppression  of 67.5, 87.5, 78.8, and 86.2 for Citrus aurantifolia, stingless bee honey, a  combination of Citrus aurantifolia-honey, and DHP, respectively. The maximum parasite suppression (87.5%) was exerted by the stingless bee honey group.  There were no significant differences in parasite suppression in the DHP group,  honey, a combination of honey and Citrus aurantifolia. Phytochemical analysis  revealed that stingless bee honey contains saponins, flavonoids, tannins,  phenols, and alkaloids, while lime contains several saponins, flavonoids, tannins,  phenols, alkaloids, and steroids. A combination of Citrus aurantifolia  and honey has potential to produce an antimalarial activity in vivo. However, the  dose of Citrus aurantifolia in this study was relatively high and the effect of  its extract on internal organs was not yet observed. Further research is required  to study to what extent lower doses of Citrus aurantifolia produce an  antimalarial activity.&nbsp

Chronic inflammation is common in infectious diseases, rheumatoid arthritis, gout, and autoimmune diseases. However, using non-steroidal anti-inflammatory drugs (NSAIDs) is accompanied by dangerous side effects. Therefore, searching for safer alternative therapies without side effects is very important. A natural blend of ingredients produced by stingless bees from plants was potential as a remedy. Meanwhile, the potential of kelulut bee products from East Kalimantan as an anti-inflammatory is still unknown. This study aimed to compare the chemical composition of kelulut bee (Trigona apicalis) products and evaluate the anti-inflammatory effect of honey, propolis, and their combination. Propolis extract and honey were determined as secondary metabolites. An anti-inflammatory in vivo assay triggered the edema using carrageenan on male mice and measured its anti-inflammatory power value. Propolis extract and honey from T. apicalis have a promising anti-inflammatory effect and are significantly higher than the positive control. Meanwhile, combining propolis extract and honey did not enhance the anti-inflammatory effect. In addition, combining honey and propolis preparations with a ratio of 75 : 25 has a better effect on reducing edema volume than the other two combinations. Still, it is not better than the treatment with propolis extract or honey alone. The content of polyphenol compounds found in honey and propolis preparations is thought to have an important role in reducing edema volume.

This review provides a comprehensive overview of stingless bee honey (SBH), focusing on its physicochemical properties, biochemical composition, and health benefits. SBH exhibits unique characteristics, including color, flavor, texture, and lower thermal stability, influenced by various botanical sources, stingless bee species, and diverse geographic and climatic conditions. These environmental factors, particularly floral diversity and climate, significantly affect SBH’s composition and functional properties, resulting in regional variations that impact its market value and health potential. The review details the composition ranges of SBH, which include moisture content (13.26%–41%), total acidity (12.59–270 meq/kg), reducing sugar (22.44%–78.95%), electrical conductivity (0.11–101.88 mS/cm), hydroxymethylfurfural (HMF) (0.21–72 mg/kg), diastase activity (0.04–16.7 mg/kg), protein (0.12%–16.5%), ash (0.07%–33%), pH (2.9–4.8), sucrose (0.01%–5.06%), proline (27–771 mg/kg), total phenolic content (0.98–854.62 mg GAE/g), total flavonoid content (2.96–279.73 mg QE/g), DPPH antioxidant activity (1.98–97.24), FRAP (0.19–62.40 mmol Fe 2+ /100 g), and trehalulose (13%–57%). SBH is rich in bioactive compounds, such as phenolics, flavonoids, enzymes, and vitamins, which may provide various health benefits. This review emphasizes findings from in vitro and in vivo studies, highlighting SBH’s antioxidant, antimicrobial, anti‐inflammatory, and wound‐healing properties. While preliminary evidence suggests potential benefits for managing diabetes, cardiovascular diseases, and gastrointestinal disorders, further research is needed to validate these claims. The lack of standardized quality requirements for SBH underscores the need for regulatory criteria to ensure product integrity in functional food markets. Establishing quality parameters and implementing a certification program for producers, alongside regular testing, will enhance product integrity and build consumer trust in the SBH market. Overall, SBH is presented as a promising yet under‐researched natural product, aiming to guide future studies and inform stakeholders in the food and health industries about its contributions to human health and wellness.

There is historical traditional use of stingless bee honey (SBH). However, no information concerning composition and antimicrobial properties of SBHs from Sudan was reported. Thus, the paper was aimed to focus on the components of the nests and honey characteristics of three species of stingless bee. Proximate analysis showed similar composition, except protein and ash were significantly (p > 0.01) different. Meliplebeia beccarii honey contained more protein 5.31% and slightly more ash 1.01% than the other species. Antibacterial activity was determined by cup-plate agar diffusion; Hypotrigona squamuligera and M. beccarii were more active against Staphylococcus aureus. None of the Studied SBHs showed activity against Escherichia coli, Apergillus niger, and Candida albicans. Anti-parasite activity as determined by sub-culture method showed that “almost all” SBHs have similar activities against trophozoites of Giardia lamblia and Entamoeba histolytica with statistically significant differences when compared to the untreated control. The amount of honey produced by M. beccarii varied from 100 to 150 ml per nest while honey produced by Hypotrigona ruspolii and H. squamuligera varied from 500 to 1000 ml/colony/year. Results also showed different nest architecture and components (entrance, brood areas, honey pots, and pollen pots) as indication for existence of many taxa of Meliponini in Sudan.

Cat's whisker leaves contain secondary metabolite compounds, flavonoids and essential oils. The chemical compounds contained in cat's whisker leaves have antioxidant, antibacterial, anti-inflammatory and antifungal properties. Flavonoids are known to function as anti-inflammatories by inhibiting pro-inflammatory enzymes, namely by blocking cyclooxygenase or lipoxygenase, can influence cellular signaling pathways such as NF-kB which can inhibit the migration of inflammatory cells to areas of inflammation. Objective: to test the anti-inflammatory effectiveness of ethanol extract of cat's whisker leaves in-vivo. Method: Anti-inflammatory effectiveness testing was carried out on male mice (Mus musculus) by inducing edema on the soles of their feet using 0.1 ml of 1% carrageenan. This research was an experimental study with 5 treatments and 5 replications. The parameters observed were edema volume, edema percentage, and edema inhibition percentage. Edema volume measurements were taken every 60 minutes for 6 hours. The results of measuring the volume of edema were used to calculate the percentage of edema, which was then used to calculate the percentage of edema inhibition. Data on the percentage of edema inhibition were statistically analyzed using the One Way ANOVA test. Result: the results of statistical tests showed significant differences in data on the percentage of edema inhibition and the percentage of edema between treatment groups showing p<0.05. From the results of the percentage of edema inhibition, it shows that cat's whisker leaf extract with a concentration of 15% has the best anti-inflammatory effect, namely inhibition of 88.42% is the concentration with the optimum dose.

Antioxidant and antibacterial properties of stingless bee (Meliponula spp.) honey from the northern highlands of Tanzania, in comparison with Apis mellifera honey

Stingless bee (Meliponini) honey has long been considered a high-value functional food, but the perceived therapeutic value has lacked attribution to specific bioactive components. Examination of honey from five different stingless bee species across Neotropical and Indo-Australian regions has enabled for the first time the identification of the unusual disaccharide trehalulose as a major component representing between 13 and 44 g per 100 g of each of these honeys. Trehalulose is an isomer of sucrose with an unusual α-(1 → 1) glucose-fructose glycosidic linkage and known acariogenic and low glycemic index properties. NMR and UPLC-MS/MS analysis unambiguously confirmed the identity of trehalulose isolated from stingless bee honeys sourced across three continents, from Tetragonula carbonaria and Tetragonula hockingsi species in Australia, from Geniotrigona thoracica and Heterotrigona itama in Malaysia and from Tetragonisca angustula in Brazil. The previously unrecognised abundance of trehalulose in stingless bee honeys is concrete evidence that supports some of the reported health attributes of this product. This is the first identification of trehalulose as a major component within a food commodity. This study allows the exploration of the expanded use of stingless bee honey in foods and identifies a bioactive marker for authentication of this honey in associated food standards.

This study aimed to determine the anti-inflammatory effect of seagrass (Enhalus acoroides) leaves on male mice (Mus musculus). The type of research used is the RAL method (completely randomized design). Male mice (Mus musculus) were divided into 5 treatment groups including group 1 being mice given 1% Na.CMC suspension as a negative control, group 2 being mice given seagrass leaf extract (Enhalus acoroides) 75 mg/kgBW, group 3 being mice. given 100 mg/kgBW of seagrass leaf extract (Enhalus acoroides), group 4 was mice given 125 mg/kgBW of seagrass leaf extract (Enhalus acoroides) and group 5 was mice given 50 mg of diclofenac sodium as a positive control. The anti-inflammatory test was induced with 1% carrageenan, the volume of edema that occurred after treatment was measured by dipping it into a plestimometer for every 30, 60, 90, 120 minute intervals for 7 days. After that, the percentage decrease in the edema volume parameter was observed. The data were analyzed using the One Way Anova test and continued with the LSD post hoc test. The ethanolic extract of seagrass (Enhalus acoroides) leaves showed that it could reduce the volume of edema in the feet of mice (Mus musculus) at concentrations of 75 mg/kgBW, 100 mg/kgBW, and 125 mg/kgBW by showing a significant difference with the negative control group (p < 0.05). The anti-inflammatory effect of the ethanolic extract of seagrass (Enhalus acoroides) leaves on male mice (Mus musculus) showed effective results in reducing the volume of edema in the legs of mice (Mus musculus).

The stingless bee honey protects against hydrogen peroxide-induced oxidative damage and lipopolysaccharide-induced inflammation in vitro

This study aimed to demonstrate the effects of moderate-intensity swimming exercise in the morning and evening over 6 weeks on the increase in growth hormone (GH) levels and the decrease in cortisol levels in male mice (Mus musculus). This research is a true-experimental study with a randomized control group posttest-only design. A total of 33 eight-week-old male mice (Mus musculus) weighing 20–40 grams were randomly divided into three groups: CN (n = 11; control without intervention), ME (n = 11; morning exercise), and AE (n = 11; afternoon exercise). Morning and evening exercises with moderate intensity were conducted for 30 minutes per session with a frequency of 3 times per week for 6 weeks. ELISA was used to evaluate posttest GH and cortisol levels in all samples. Data analysis was performed via one-way ANOVA, followed by Tukey’s honestly significant difference (HSD) post hoc test, with a significance level of p ≤ 0.05. The results revealed differences in the increase in GH levels between ME vs CN (5.71±0.88 vs 2.54±0.21 ng/mL, p=0.001), AE vs CN (7.18±1.66 vs 2.54±0.21 ng/mL, p=0.001), and AE vs ME (7.18±1.66 vs 5.71±0.88 ng/mL, p=0.009). Similarly, the decrease in cortisol levels differed among the ME vs CN (6.62±2.37 vs 10.69±4.38 ng/mL, p=0.004), AE vs CN (2.18±0.99 vs 10.69±4.38 ng/mL, p=0.001), and AE vs ME (2.18±0.99 vs 6.62±2.37 ng/mL, p=0.008) groups. This study revealed that both exercise durations significantly increased growth hormone levels and decreased cortisol levels in male mice (Mus musculus). However, evening exercise with moderate intensity was more effective at increasing growth hormone levels and decreasing cortisol levels than morning exercise with moderate intensity in male mice (Mus musculus). Keywords: Growth hormone, cortisol, morning exercise, afternoon exercise, metabolism

The local stingless bee honey in Malaysia is highly valued because of its significant nutritional and natural remedy. Two commonly found stingless-bee species in Malaysia are Heterogona itama and Geniotrigona thoracica. Currently, the lack of standard profile data for the pure stingless bee honey from these two species leads to challenges in evaluating the authenticity of the stingless bee honey. To address this gap and provide a reliable solution, the objective of this research work is to develop a smart profiling system that can characterize stingless bee honey. This research work presents an innovative 3-in-1 smart profiler for characterizing pH, moisture content and colour intensity for different concentrations of stingless-bee honey. The developed profiler can characterize the stingless bee honey profile in situ without tediously bringing the samples to the laboratory. The 3-in-1 smart profiler utilized an Arduino Uno microcontroller, a TCS3200 colour sensor, an E-201-C pH sensor and an HD-38 moisture sensor. The research methodology involves designing the simulation environment using Tinkercad software, developing a hardware prototype, system testing, sensor calibration and performing comprehensive analysis. The developed profiler has successfully characterized pH level, moisture content and colour intensity for 100% pure honey, 75%, 50%, 25%, and 12% stingless bee honey with the repeatability of the output trend. For Heterogona itama species, the measured acidity level, moisture content and RGB colour intensity for 100% pure honey are in the range of 2.76-2.77, 82-84 % and 973-976-726 respectively. On the other hand, 100% pure honey from Geniotrigona thoracica species has 2.4-2.42 pH level, 86-89 % moisture content and RGB values of 970-963-707. This smart profiling system has the potential to be a part of the profiler in creating a standard profile for stingless bee honey for itama and thoracica. This research work has advantages in terms of in-situ detection, affordable to own and simple operating sensing system. It has the potential to be marketed and promote local socio-economic development. In the future, the accuracy of the results obtained from the prototype will be compared to sophisticated equipment such as gas chromatography.

The demand for natural products that promote sustainable production chains is growing worldwide. Honey, while not a staple for food security due to its limited nutritional contribution, represents a valuable resource to foster dietary diversity, responsible consumption, and forest conservation. Its production depends directly on access to diverse flora with low pesticide exposure and requires minimal soil transformation or external inputs, making it a low-impact practice. Beekeeping also contributes to the maintenance of pollinator populations, which are key for biodiversity and sustainable agriculture. Consumer interest in the origin and authenticity of honey has grown in recent years, especially for Apis mellifera products (Escuredo and Seijo 2024, Mascarello et al. 2024). Less known is the diversity and economic potential of stingless bees (Meliponini). Across tropical and neotropical regions, nearly 500 species have been described (Grüter 2020), many of which produce honey and act as key pollinators. In Colombia, about 34 species are reportedly used in meliponiculture (Nates-Parra and Rosso-Londoño 2013). This practice is expanding in rural areas, where it complements agriculture and is compatible with family-centered management. Traditional knowledge attributes medicinal properties to stingless bee products (Sgariglia et al. 2010, Rosales 2012). Although clinical validation remains limited, biochemical studies of their honeys have identified distinctive compositions rich in phenolic compounds and other bioactive molecules, providing preliminary evidence of antioxidant and anti-inflammatory properties that warrant further research (Gomes et al. 2022). These properties are linked to microbial communities naturally present in stingless bee honeys. Fermentative yeasts and lactic acid bacteria shape these products, creating unique ecological niches with potential probiotic and bioactive value (da Silva et al. 2024). Microbial assessment is also essential to detect possible pathogens, as not all stingless bee species are recommended for meliponiculture: Trigona species, for example, may incorporate feces or dead animal tissue into nests, raising sanitary concerns (Gómez et al. 2023) Developing markets for stingless bee products requires robust methodologies for authentication and safety. We propose environmental DNA (eDNA) as a tool to address three dimensions of traceability and diversity assessment: Entomological origin of honey. Despite vast bee diversity, taxonomic expertise has declined, limiting our ability to monitor wild bees. DNA-based approaches, particularly metabarcoding, enable verification of the bee taxa involved in honey production—critical in regions with limited expertise. Challenges include primer selection for non- Apis bees, expanding reference databases, and training in molecular methods. Incorporating eDNA and metabarcoding into product traceability could democratize biodiversity monitoring and inform public policy. Botanical foraging spectrum. DNA from bee pollen and honey reveals floral visitation patterns, offering a proxy for ecosystem health. Progress is constrained by incomplete plant reference libraries. Diversity indices—borrowed from microbiome studies—can help characterize floral communities, though interpretation must account for variable foraging behavior across bee species. Microbial diversity and bioprospecting. Stingless bee honeys act as natural “filters,” concentrating microbial communities with industrial, probiotic, or antibiotic potential. Although generally safe, rigorous biosafety evaluation is needed to balance opportunities with risks. Reliable isolation and cultivation methods will be key to characterize diversity, establish reference libraries, and develop pipelines for future bioprospecting. Entomological origin of honey. Despite vast bee diversity, taxonomic expertise has declined, limiting our ability to monitor wild bees. DNA-based approaches, particularly metabarcoding, enable verification of the bee taxa involved in honey production—critical in regions with limited expertise. Challenges include primer selection for non- Apis bees, expanding reference databases, and training in molecular methods. Incorporating eDNA and metabarcoding into product traceability could democratize biodiversity monitoring and inform public policy. Botanical foraging spectrum. DNA from bee pollen and honey reveals floral visitation patterns, offering a proxy for ecosystem health. Progress is constrained by incomplete plant reference libraries. Diversity indices—borrowed from microbiome studies—can help characterize floral communities, though interpretation must account for variable foraging behavior across bee species. Microbial diversity and bioprospecting. Stingless bee honeys act as natural “filters,” concentrating microbial communities with industrial, probiotic, or antibiotic potential. Although generally safe, rigorous biosafety evaluation is needed to balance opportunities with risks. Reliable isolation and cultivation methods will be key to characterize diversity, establish reference libraries, and develop pipelines for future bioprospecting. Our pilot project, in the Magdalena Medio region of Colombia, applies eDNA analyses to bees, honey, pollen, and associated microorganisms. The study focuses on three genera of stingless bees maintained in boxes derived from colonies collected in situ in the El Silencio nature reserve of Fundación Biodiversa Colombia, ensuring foraging in a pesticide-free environment with high floral diversity. As one of the first efforts to apply DNA-based tools to stingless bee biodiversity surveys and product authentication, this initiative highlights the need for stronger collaboration, standardized data frameworks, and investment to unlock the potential of eDNA for conservation and bioeconomic development.

There were about 500 stingless bee species that had been reported in Afrotropical, Indo-Australian and Neotropical provinces with the highest diversity species originating in the Neotropical. In Malaysia, there are more than 38 species and around 33 species were reported in Peninsular Malaysia. Due to its high nutritious and medicinal values, recently, stingless bee honey has been highly demanded by the food, pharmaceutical and even cosmetic industries. Thus, this paper describes the cosmeceutical potential of stingless bee honey as an antiacne, skin moisturizing and anti-hypertrophic scar agent. Stingless bee honey can treat acne by reducing inflammation and irritation during acne formation due to its high flavonoid and phenolic compounds. Besides, stingless bee honey also possesses antibacterial activity due to its acidity, osmolarity and hydrogen peroxide content which is unfavourable for P.acne growth. It functions as a natural humectant due to its ability to attract water from the dermis and deeper epidermis to the epidermis and its high water-binding capacity. Stingless bee honey also can reduce scar formation by improving the wound healing process. This is because it can possess both pro-inflammatory and anti-inflammatory properties in which it triggers the production of the pro-inflammatory cytokines to initiate the inflammation and inhibit the production of the pro-inflammatory cytokines when inflammation is in progress to avoid prolonged inflammation. Also, stingless bee honey can stimulate skin reepithelialization and wound contraction. These will reduce the damaged tissue that needs to be repaired and eventually minimize the scarring area. Hence, due to all of these benefits, further research can be done to study the action, use and effectiveness of stingless bee honey in cosmetic applications.

Graphical Abstract Research Highlights The combination of honey and hirta improved hematological parameters of C. gariepinus catfish infected with A. hydrophila bacteria. The combination of honey and hirta positive affected the histopathology of liver, kidneys, and gills, which led to a restoration of organ function to normal levels. The combination of honey and hirta also improved the morphology of C. gariepinus. The most effective combination of honey and hirta was treatment C (1:2). Abstract Honey and Euphorbia hirta are natural ingredients that may serve as alternative treatments for Clarias gariepinus catfish infected with Aeromonas hydrophila bacteria. Both honey and E. hirta contain bioactive compounds with known antibacterial properties. This study aims to investigate the effects of combining honey and E. hirta on various hematological parameters, including erythrocytes, leukocytes, MCH, MCV, as well as histopathological and morphological changes in C. gariepinus catfish infected with A. hydrophila. This study used an experimental design involving three treatment combinations of honey and E. hirta, which were A (2:1), B (1:1), C (1:2) and a control with three replications. The treatments were administered to the fish through immersion. Honey was diluted at a 50% concentration in distilled water. Meanwhile, E. hirta was prepared as a coarse powder and boiled at a 3% concentration. The results showed that the combination of honey and E. hirta affected the hematological, histopathological, and morphological parameters of C. gariepinus catfish within the normal ranges. The most effective treatment was found to be treatment C (1:2). Therefore, the combination of honey and E. hirta is potential as a therapeutic option for bacterial infections in fish.

BACKGROUND: Cancer growth is influenced by many factors and in general it is an interaction between gene factors and environmental factors, especially the microenvironment that exists around cancer. The inflammatory response plays a decisive role in various stages of cancer growth. AIM: The aim of this study was to determine the anti-inflammatory activity of ethanol extract of Rhaphidophora pinnata leaves. METHODS: R. pinnata leaf extract was obtained by percolation method using 96% ethanol as the solvent at room temperature. Anti-inflammatory activity was determined based on the paw edema method. Thirty male albino mice were treated orally with sodium carboxyl cellulose suspension (as negative control group), R. pinnata leaf extract (35, 70, 140, and 280 mg/kgBW), and diclofenac (as positive control group), 60 min before 0.2 mL 1% carrageenan injection. The paw thickness was measured using plethysmometer before injecting the carrageenan and after 1, 2, 3, 4, 5, and 6 h. RESULTS: The subplantar injection of carrageenan caused a time-dependent paw edema in the mice. Oral administration of R. pinnata leaf extract inhibited paw swelling at 1, 2, 3 4, 5, and 6 h after carrageenan injection. R. pinnata leaf extracts doses of 35, 70, 140, and 280 mg/kgBW gave a percentage inhibition of 56.56%, 56.18%, 62.77%, and 49.30%, respectively. The effective dose of R. pinnata leaf extract as an anti-inflammatory was 140 mg/kgBW. CONCLUSION: Ethanol extract of R. pinnata leaf has anti-inflammatory activity in male albino mice.