Abstract Alzheimer's disease (AD) is currently a global concern, and treating its current symptoms and slowing its progression is an important goal. To this end, identifying biochemical mechanisms that have the potential to prevent and improve the pathology of the disease is a priority. The response of the central nervous system to pathological insults may be neuroinflammation. Kynurenine (KYN) pathway metabolites, which are closely related to neuroinflammation, may have the potential to affect many neural processes such as synaptic plasticity, neurotransmission, antioxidant mechanisms, and neurogenesis. In addition, it has been recently shown that there is a strong correlation between sensory system deterioration and AD progression and that neuropathological changes in sensory regions can be detected before changes in memory-related regions. For this purpose, our study aimed to investigate the effects of changes in the Kynurenine (KYN) pathway on cognitive functions and auditory sensory system functions in an experimental AD model and the effects of bee bread (BB), known to have anti-inflammatory properties, on these parameters. It was shown that BB has the potential to improve memory by exerting a healing effect on neuroinflammation, apoptosis, and the KYN pathway, and also has a therapeutic effect on auditory sensory system functions, reducing sensory system integrity disorders. These results highlight the ability of BB to prevent neurotoxic effects associated with sAD. It may be a potential protector as a food supplement for AD neuropathological health, and further studies are needed for this purpose. Graphical Abstract

Determination of pesticides, antibiotics, PAHs, PCBs, and plasticizers in honeybees, honey, and other bee products - modified QuEChERS method. Method review for 2018-2024.

Bee pollen is commonly consumed as a natural dietary supplement due to its rich nutritional content and bioactive components. However, consuming it directly without any processing steps can pose a potential risk to public health concerning microbiological safety. This study aimed to assess the microbiological quality and certain physicochemical properties of bee pollen available for sale in the Şanlıurfa region. In this context, a total of 12 different bee pollen samples were analyzed, collected from local herbalists and apiculture product sales outlets. In the physicochemical analyses, pH, moisture, and color (CIE Lab*) values were determined using standard analytical methods. For the microbiological analyses, counts were conducted for total mesophilic aerobic bacteria (TMAB), coliform bacteria, Escherichia coli, Enterobacteriaceae, yeast, and mold. Additionally, analyses were performed to detect the presence of E. coli O157:H7, Salmonella spp., Listeria monocytogenes, and Staphylococcus aureus. The pH, moisture, and color (CIE Lab*) values were found to be 4.24–5.33 (average 4.91 ± 0.29), 9.87–24.66% (average 14.18 ± 4.40), and L* 23.30 ± 4.27, a* 10.81 ± 3.30, b* 27.43 ± 7.44, respectively. In the microbiological analyses, total mesophilic aerobic bacteria (TMAB) counts ranged from 4.38 to 5.28 log CFU/g, yeast–mold counts from 2.34 to 3.92 log CFU/g, coliform bacteria from 1.97 to 3.27 log CFU/g, and Enterobacteriaceae from 1.73 to 4.23 log CFU/g. E. coli, S. aureus, Salmonella spp., L. monocytogenes, and E. coli O157:H7 were not detected in any of the samples. These results indicate that bee pollen is safe in terms of pathogens, but mold development in some samples may pose a mycotoxin risk. In conclusion, it was determined that bee pollen sold in the Şanlıurfa region is generally of suitable quality but requires attention regarding microbiological safety. To consider bee pollen a safe functional food, it is necessary to improve hygiene conditions during production and storage processes and to establish microbiological quality standards.

Objective: This study aimed to evaluate the protein extraction efficiency and amino acid composition of bee pollen collected from different botanical sources using water-based ultrasonic-assisted extraction (UAE) with various solvents. Material and Methods: Bee pollen samples were collected from five different apiaries in the central district of Bingöl, Türkiye. Palynological analysis was conducted to identify botanical origins, followed by UAE using distilled water (dH₂O), ethanol (EtOH), and methanol (MeOH) at different concentrations. Extraction efficiency and protein content were determined, and the amino acid profile was analyzed in the sample with the highest protein content. Results: Palynological analysis showed diverse floral sources across the samples. The highest extraction yield (75%) was obtained from the BP-1 sample using 25% EtOH, while the highest protein content (9.80%) was observed in BP-3 with dH₂O. BP-3, primarily derived from Fabaceae, Poaceae, and Polygonaceae, was rich in proline, asparagine, and glycine. Conclusion: The botanical origin of bee pollen significantly affects its chemical composition. Solvent type and concentration critically influence extraction efficiency and protein yield. UAE with dH₂O offers a practical method for obtaining plant-based protein from bee pollen, contributing to its valorization as a functional food source. Keywords: Bee Pollen, Ultrasonic-Assisted Extraction (UAE), Protein Content, Amino Acid Composition

Acorn bee pollen is a highly nutritious natural food, but information regarding its quality properties is currently lacking. Therefore, in this study, acorn bee pollens were collected from five regions of South Korea, and then acorn bee pollen extracts (ABPe) were prepared, and their quality properties, as well as their components, were analyzed. For the analysis related to the ABPe’s antioxidant activity, the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical, 2,2-diphenyl-1-picrylhydrazyl radical, and nitrite scavenging activities of JNe (sample from Jeonnam) were at 72.7%, 66.8%, and 60.0%, respectively, which were significantly higher than their respective averages of 69.7%, 56.7%, and 55.3% for the other four samples. Moreover, the inhibition activities of JNe on tyrosinase, α-glucosidase, and α-amylase were 72.5%, 92.6%, and 72.4%, respectively, which were significantly higher than their respective averages of 62.7%, 70.9%, and 61.5% for the other four samples. When considering the ABPe components, the total flavonoid content of JNe was 8.4 mg QE/g, which is higher than the average of 4.6 mg QE/g for the other four samples, and exhibited the highest correlation with the quality properties of acorn bee pollen. Furthermore, among the quercetin glycosides in JNe, rutin revealed a higher correlation with the quality properties than the other flavonoids. Therefore, total flavonoid and rutin content are closely associated with the antioxidant activity and enzyme inhibition of the ABPe. In conclusion, these results are expected to be used for evaluating the quality properties of acorn bee pollen as a healthy functional food ingredient or a natural food additive.

Bee bread as a natural shield against genetic damage: Investigation of antigenotoxic and antimutagenic potential.

The supplementary feeding of bee colonies under conditions of limited natural food sources is essential for maintaining their health and productivity. Pollen is a major source of protein for bees, collected during plant flowering. Its absence negatively affects the secretory activity of the hypopharyngeal glands (HPGs) in young nurse bees. This study aimed to assess the effect of different protein sources provided during the spring period on the development of HPGs in worker bees. An experiment was conducted with 28 bee colonies divided into seven groups of four colonies each—one control and six experimental. The colonies were fed soy isolate (Glycine max), brewer’s yeast (Saccharomyces cerevisiae), spirulina (Arthrospira platensis), pea protein (Pisum sativum), and bee pollen (a mixture of pollens from various plant species). The results confirmed the critical role of pollen, as 40% of bees in the pollen-fed group exhibited HPGs at grade 4. The group supplemented with spirulina showed similar results (35.8%), indicating its potential as an alternative protein source. These findings highlight the importance of proper protein supplementation and suggest that spirulina could serve as a promising substitute for pollen in early spring feeding to support colony development.

Honey bees play a vital role in both the production of high-value hive products and the pollination of a wide range of crops, making them indispensable to global agriculture. However, during dearth periods when natural sources of nectar and pollen are limited, colony strength often declines, threatening brood rearing and overall productivity. Nutritional supplementation through pollen substitute diets has emerged as an important strategy to sustain colonies under such conditions. To evaluate their effectiveness, experiments were conducted during the dearth seasons of 2021 and 2022 at the apiary of the Horticultural Research Farm, Chaudhary Charan Singh Haryana Agricultural University, Hisar. Pooled results from both years revealed that treatment T10 (Bee Pollen + Honey) produced the highest mean colony strength (8.21 frames), demonstrating a significant positive impact on brood development and foraging activity. This superior performance was attributed to the synergistic nutritional benefits of combining natural pollen with honey. The second-best results were obtained with treatment T7 (a formulated diet of dehusked parched gram flour, deactivated yeast, skimmed milk powder, sugar, water, turmeric powder, multivitamins, and pollen), which achieved a mean strength of 7.48 frames. This was followed by T6 (T3 + 10% Bee Pollen) with a mean of 7.20 frames, suggesting that the inclusion of yeast, turmeric, and multivitamins enhanced colony vitality, possibly by boosting immunity and resilience. Overall, the findings highlight that carefully formulated pollen substitute diets can effectively maintain colony strength during forage scarcity, with Bee Pollen + Honey proving to be the most beneficial option.

Pollen grains are being explored as innovative biomaterials for different applications, ranging from oral drug delivery to encapsulation of food additives, with the production of pollen-based building blocks standing on its robust, chemically inert, and mechanically durable sporopollenin wall. Yet, concerns remain regarding the safety of sporopollenin microcapsules (SMCs) or derivatized sporopollenin materials purified from pollen grains, traditionally linked to allergies. Herein, we address the critical question of whether sporopollenin shells purified from bee pollen may cause allergic reactions by evaluating their interaction with human immunoglobulin E (IgE) antibodies in sera from patients with and without allergic sensitization to pollen of specific species. Clean SMCs fromCastanea sativa, Amaranthaceae (Chenopodium album), andOlea europaeapollen grains were successfully produced using a species-independent chemical treatment and characterized. The Covaris Adaptive Focused Acoustics™ (AFA) technology was employed for protein extraction from the bee pollen and the SMCs, yielding 0.72-1.25 ng and 0.026 ng-0.028 ng of protein per pollen grain, respectively. X-ray photoelectron spectroscopy (XPS) analysis also confirmed that the surface nitrogen content of the SMCs was minimal, ranging from 0.9% to 2.7%. SDS-PAGE, followed by immunoblotting analysis, showed that proteins extracted from bee pollen strongly reacted with IgE antibodies in human sera, whereas SMCs did not trigger allergic sensitization. Overall, our findings suggest that while bee pollen proteins could elicit allergic reactions in sensitive patients, SMCs do not, highlighting their potential as safe biomaterials for various applications and offering valuable insights into the allergenic potential of bee pollen.

Bee pollen is a food with high nutritional value and important functional properties. It is usually consumed as dried pollen, due to the need for a preservation process that controls the high microbiological load of the fresh product. Unfortunately, dry pollen is unattractive to consumers. In this sense, the use of ozone may be an alternative for preserving fresh pollen, since it reduces the microbiological load, allowing for other preservation methods, including refrigeration, and it preserves the original texture and flavors of the product, making it more palatable. However, it is important to know how ozone can affect some of the bioactive properties of pollen, such as phenolic compounds (PC), or the antioxidant properties of this food. The aim of this research was to assess the effect of ozone treatment on the above-mentioned properties and to compare it with the conventional drying treatment. For this purpose, 19 samples of fresh bee pollen were collected. From each sample, five subsamples were obtained, two of which were treated with ozone for 1 h and 2 h (O1 and O2, respectively), two were dried for 4 h and 8 h (D4 and D8, respectively), and the fifth subsample remained as the untreated control (C). The results showed that ozone treatments did not have a negative effect on phenolic content (PC) or on antioxidant activity (AA). This would be positive for the use of this decontamination method. In contrast, traditional drying treatments significantly reduced total PC while increasing AA compared to C, O1, and O2. A low correlation was found between these variables in the studied samples.

Bee pollen is a natural nutrient substance collected by bees from plants. Its metabolites have been extensively studied, yet the characteristic metabolites of bee pollen from different floral sources have not been clearly identified. In this study, we collected four types of bee pollen (tea, rose, rapeseed, and corn pollen) from across China and analyzed their volatile and non-volatile metabolites using liquid chromatography-high-resolution mass spectrometry (LC-HRMS) and gas chromatography-mass spectrometry (GC-MS). At the same time, the nutritional substances (Including polyphenols, organic acids, and sugars) were precisely quantified. The results showed that the total phenols (5 mg GAE/g) and total flavonoids (0.27 mg RE/g) content of corn pollen were significantly higher (p < 0.05) than those of other pollens, and the contents of polyphenols such as naringenin were relatively high, indicating strong antioxidant potential. Rose pollen was rich in protein (0.04 g/g) and flavonoid glycosides. Tea pollen was prominent in the content of polyphenol glycosides and amino acid derivatives, while rapeseed pollen performed well in phenolic acids (Ferulic acid), as well as specific sugar (Mannose). We identified the differential metabolites of these bee pollen through orthogonal partial least squares discriminant analysis (OPLS-DA) (VIP > 1). It was also stipulated that metabolites with a VIP value greater than 1.5 showed significant differences and could be used as characteristic metabolites for differentiating pollen (p < 0.05). The representative metabolites of bee pollen were as follows: rapeseed pollen—ferulic acid; tea pollen—malic acid; corn pollen—epicatechin; and rose pollen—fumaric acid. This study provides a research basis for evaluating the quality, traceability, and metabolite exploration of bee pollen.

Bee pollen and propolis are commonly used for therapeutic and prophylactic purposes in both humans and animals. This study evaluated their effects on the morphology of central lymphoid organs in broiler chickens. Birds were fed fodder supplemented with 0.025% propolis and/or 0.5% bee pollen during the first two weeks of fattening. Despite a natural, asymptomatic Salmonella enteritidis infection, no significant structural differences were observed in the thymus, spleen, or bursa of Fabricius. However, experimental groups - particularly those receiving propolis or bee pollen - showed signs of enhanced lymphoid activity and epithelial development. These findings suggest a protective and immunostimulatory effect of bee products on lymphoid organs, even during infection.

Abstract Bacteria are spread throughout all parts of the human body, including the human oral cavity. One common problem related to the oral cavity is dental caries. As much as 93% of Indonesian children experience dental caries issues. Streptococcus mutans and Streptococcus sobrinus are bacteria that play a role in dental caries. Bee bread is a source of nutrition for bee colonies and serves as food for larvae and young bees. The study aims to determine the effectiveness of bee bread extract on inhibiting bacteria Streptococcus mutans and Streptococcus sobrinus. The bee bread used comes from the Kendilo area, Penajam Paser Utara, East Kalimantan. The crude extract of bee bread is dried and then macerated also concentrated using a rotary vacuum evaporator. Antibacterial testing was conducted using Mueller Hinton Agar (MHA) media with chloramphenicol as the positive control. The test results show that the bee bread extract of Heterotrigona itam a is effective in inhibiting Streptococcus mutans and Streptococcus sobrinus bacteria with strong antibacterial activity on both types of bacteria.

Abstract Heterotrigona itama is one of the stingless bee species with high potential for honey production and is widely cultivated in East Kalimantan, particularly in Samarinda. However, its domestication often faces challenges due to limited knowledge of forage management and the influence of microenvironmental factors on productivity. This study aims to examine the effects of floral resource availability and environmental conditions on the daily foraging activity, colony structure, and the productivity of honey and bee bread of H. itama in two meliponiaries in Samarinda. Data were collected from six colonies through direct observation of foraging behavior, pollen identification using the acetolysis method, and measurement of microclimatic parameters including temperature, humidity, light intensity, and wind speed. The results indicated that foraging activity followed a diurnal pattern, peaking at 11:00 a.m., with light intensity showing a very strong positive correlation with nectar collection activity (r = 0.95), while humidity was negatively correlated. A total of 108 plant taxa from 44 families were identified in both honey and bee bread samples, indicating that H. itama is a multifloral species. Wilcoxon tests revealed significant differences (p &lt; 0.05) in pollen composition between the two sites. Honey and bee bread productivity were positively correlated with the proportion of worker bees (r = 0.65 and r = 0.72, respectively), and several plant species such as Nuxia sp. and Capsicum sp. were found to significantly contribute to production output. This study highlights that proper floral resource management and microclimate regulation are key factors in enhancing the productivity of stingless bees in tropical regions.

Abstract Beehive products have been known since ancient times and are used in medicine, food, and beverages. Honey, bee bread, propolis, pollen, royal jelly, and beeswax are valuable sources of active compounds that contribute to modern apitherapy and help combat civilization-related diseases. However, there is growing attention to the microbiota of beehive products, which vary among different products and depend on several factors, such as the environment surrounding the hive, the collection process of bee products, and their subsequent processing. Culture-dependent and culture-independent methods have been used to investigate the microbiota of bee products. Commonly identified microorganisms include lactic acid bacteria, spore-forming Bacillus species and related genera, osmophilic yeasts, and xerophilic molds. The microbiota of bee products holds underexplored potential for applications in industry and medicine, including as potential probiotics, sources of antimicrobial agents, and enzyme producers.

Abstract This study investigates the diversity of forage plants and the pollen morphology associated with Apis dorsata in mangrove stands of the Mahakam Delta. The research applied the acetolysis method to analyze bee bread samples. From these samples, five plant families comprising seven species were identified, with pollen from Rhizophora apiculata found to be the most frequently collected by A. dorsata foragers. The pollen grains were generally medium to large in size, exhibiting oblate spheroidal to prolate spheroidal forms. They occurred as monads, with apertures categorized as tricolporate, inaperturate, and tricolpate. The surface ornamentation was characterized by scabrate, verrucate, and perforate textures.

Abstract Honeybees play a crucial ecological and economic role by pollinating both natural ecosystems and cultivated crops. Beyond this, they produce such valuable natural products as honey, propolis, royal jelly, bee pollen, beeswax, and bee venom, which have been used for centuries in traditional medicine. With the growing interest in natural compounds for disease prevention and treatment, apitherapy and bee-derived products have gained renewed attention. Among these, bee venom (BV) stands out due to its rich composition of bioactive molecules, including peptides such as melittin, phospholipase A2 (PLA2), apamin, mast cell degranulation peptide, secapin, and adolapin, as well as enzymes as hyaluronidase. These components are known for their therapeutic potential, particularly in inflammation and cancer-related applications. In this study, the cytotoxic and apoptosis-inducing effects of Apis mellifera L. venom samples collected from fifty-one locations across Türkiye were tested against seven human cancer cell lines (Caco-2, PC3, U-87MG, MDA-MB-231, A549, HeLa, Panc-1) and one healthy line (CCD-34Lu). Cytotoxicity was assessed with use of the MTT assay, and IC 50 values ranged from 1.64±0.70 to 98.33±9.28 µg/ml. Venoms from Maçahel, Faraşin, Kırklareli, Düzce, Adana, Hatay, and Gökçeada were the most potent, influenced by bee ecotypes and protein content. PC3, PANC-1, and HeLa were the most sensitive cell lines, while Caco-2 and CCD-34Lu showed the highest resistance. In conclusion, the diverse venoms of A. mellifera in Türkiye exhibit significant dose-dependent cytotoxic and apoptotic effects, positioning the country as a valuable source in venom-based therapeutic research.

Addressing chemical risks in beekeeping products: development of a combined ultrasound-assisted extraction and QuEChERS-based method for the simultaneous determination of mycotoxins and pesticides in bee pollen and honey

Background: The expansion of oil palm plantations in Indonesia drives deforestation and biodiversity loss, threatening pollinator communities. Agroforestry systems within oil palm landscapes may enhance floral resources, but their effectiveness is poorly understood. This study evaluates the potential of stingless bee (Heterotrigona itama) pollen analysis as a tool for monitoring floral diversity in different oil palm agroforestry systems. Methods: Pollen samples were collected from H. itama colonies in five distinct oil palm agroforestry systems (block-type, intercropping, row-type, fence-type, and palm oil-livestock) across South Kalimantan. Pollen types were identified microscopically, and diversity metrics (Shannon Index, Simpson Index, and Evenness) were calculated to assess foraging patterns and floral resource availability. Results: Analysis identified 30 plant species from 22 families. Agroforestry type significantly influenced pollen composition. Block-type systems exhibited the highest species richness (14 types), while fence-type systems showed the lowest (9 types) with heavy dominance by maize (Zea mays, 41.4%). Intercropping systems were dominated by Acacia mangium (51.2%). Row-type systems demonstrated the most balanced foraging (highest evenness). A moderate Sørensen similarity index (0.63) between pollen and field vegetation confirmed bees as effective samplers of landscape floral diversity. This research confirms that H. itama pollen analysis is an efficient bioindicator tool, revealing that agroforestry management directly shapes pollinator resource availability. Complex systems (e.g., block-type, row-type) support higher floral diversity, which is crucial for pollinator conservation and sustainable productivity in oil palm-dominated landscapes. This method provides a practical approach for guiding biodiversity-inclusive agricultural practices.

The characteristics of bee bread microparticles from East Kalimantan's Kelulut bee (Heterotrigona itama) and their potency as antimicrobials, antioxidants, and anti-inflammatories