Clinical complications in envenoming by Apis honeybee stings: insights into mechanisms, diagnosis, and pharmacological interventions.

Envenoming resulting from Apis honeybee stings pose a neglected public health concern, with clinical complications ranging from mild local reactions to severe systemic manifestations. This review explores the mechanisms underlying envenoming by honeybee sting, discusses diagnostic approaches, and reviews current pharmacological interventions. This section explores the diverse clinical presentations of honeybee envenoming, including allergic and non-allergic reactions, emphasizing the need for accurate diagnosis to guide appropriate medical management. Mechanistic insights into the honeybee venom's impact on physiological systems, including the immune and cardiovascular systems, are provided to enhance understanding of the complexities of honeybee sting envenoming. Additionally, the article evaluates emerging diagnostic technologies and therapeutic strategies, providing a critical analysis of their potential contributions to improved patient outcomes. This article aims to provide current knowledge for healthcare professionals to effectively manage honeybee sting envenoming, thereby improving patient care and treatment outcomes.

Seasonal detection of pathogens in honeybees kept in natural and laboratory conditions.

The honeybee is one of the most important pollinators in the world. The frequently observed poor health of honeybee colonies can be caused by various factors, e.g. environmental pollution, nutritional stress, and climate changes. Moreover, honeybees are constantly exposed to a wide spectrum of pathogens, such as parasites, bacteria, and viruses. We examined the occurrence of various diseases in different-aged worker honeybees from two colonies kept in natural and laboratory conditions during spring, summer, and autumn in Poland. The honeybees were examined by PCR to detect infection with selected pathogens: Nosema ceranae, N. apis, N. bombi, Acarapis woodi, trypanosomatids, and neogregarines (Mattesia or Apicystis species) and by RT-PCR to identify deformed wing virus (DWV), black queen cell virus (BQCV), and acute bee paralysis virus (ABPV). DWV and N. ceranae turned out to be the dominant pathogens. Trypanosomatids and BQCV were also found in several samples. We did not detect the presence of the other pathogens: N. apis, N. bombi, A. woodi, neogregarines, or ABPV. As shown in the present study, the dynamics and occurrence of pathogens are influenced by keeping conditions, honeybee age, and seasonality.

Nosema ceranae infection reduces the fat body lipid reserves in the honeybee Apis mellifera.

Nosema ceranae is an intestinal parasite frequently found in Apis mellifera colonies. This parasite belongs to Microsporidia, a group of obligate intracellular parasites known to be strongly dependent on their host for energy and resources. Previous studies have shown that N. ceranae could alter several metabolic pathways, including those involved in the nutrient storage. To explore the impact of N. ceranae on the fat body reserves, newly emerged summer bees were experimentally infected, and we measured (1) the lipid percentage of the abdominal fat body at 2-, 7- and 14-days post-inoculation (p.i.) using diethyl ether lipid extraction, (2) the triglyceride and protein concentrations by spectrophotometric assay methods, and (3) the amount of intracellular lipid droplets in trophocytes at 14- and 21-days p.i. using Nile Red staining. Comparing the three methods used to evaluate lipid stores, our data revealed that Nile Red staining seemed to be the simplest, fastest and reliable method. Our results first revealed that the percentage of fat body lipids significantly decreased in infected bees at D14 p.i. The protein stores did not seem to be affected by the infection, while triglyceride concentration was reduced by 30% and lipid droplet amount by 50% at D14 p.i. Finally, a similar decrease in lipid droplet reserves in response to N. ceranae infection was observed in bees collected in fall.

[Research Progress in Detection of Bee Venom Allergens].

Hymenopteran insect stings are a risk factor that cannot be ignored for the people allergic to hymenopteran venoms.In China,the current diagnostic tools cannot provide accurate information to identify sensitized insects,thus affecting clinical diagnosis and treatment.Honeybee is a common hymenopteran insect.Due to its wide distribution,large number,and complex venom composition,researchers have carried out recombination schemes for the main allergens of honeybee venom,laying a theoretical foundation for the detection of allergens.The development of diagnostic technologies for allergen components can accurately detect bee venom allergens,providing a new set of clinical diagnosis and treatment schemes for the population allergic to bee venom.

Evaluation of Prebiotic and Health-Promoting Functions of Honeybee Brood Biopeptides and Their Maillard Reaction Conjugates.

This study addresses the growing interest in natural functional ingredients by evaluating the prebiotic and health-promoting functions of honeybee brood biopeptides (HBb-Bps) and their conjugates. The purpose was to investigate their antioxidant activities, enzyme inhibition properties, and effects on probiotic growth and short-chain fatty acid (SCFA) production. The HBb-Bps were conjugated with honey, glucose, and fructose via the Maillard reaction. Antioxidant activities were assessed using DPPH and ABTS assays. The inhibitory effects on amylase, pancreatic lipase, and the angiotensin-converting enzyme (ACE) were measured. Probiotic growth and SCFA production were evaluated using L. plantarum TISTR846, and L. lactis TISTR1464. The HBb-Bps and their conjugates exhibited enhanced antioxidant activities post-Maillard reaction. They showed moderate enzyme inhibition, which decreased after conjugation. However, ACE inhibition increased with conjugation. The HBb-Bps significantly promoted probiotic growth and SCFA production, with further enhancement by the Maillard reaction. Overall, the HBb-Bps and their conjugates demonstrate significant prebiotic and health-promoting functions, suggesting their potential as natural ingredients in functional foods and nutraceuticals. Further research should focus on the in vivo effects and, given their solubility and stability these biopeptides could be incorporated into functional food formulations, such as health beverages, protein bars, and other fortified foods designed to deliver specific health benefits.

High-resolution proteomics and machine-learning identify protein classifiers of honey made by Sicilian black honeybees (Apis mellifera ssp. sicula).

Apis mellifera ssp. sicula, also known as the Sicilian black honeybee, is a Slow Food Presidium that produces honey with outstanding nutraceutical properties, including high antioxidant capacity. In this study, we used high-resolution proteomics to profile the honey produced by sicula and identify protein classifiers that distinguish it from that made by the more common Italian honeybee (Apis mellifera ssp. ligustica). We profiled the honey proteome of genetically pure sicula and ligustica honeybees bred in the same geographical area, so that chemical differences in their honey only reflected the genetic background of the two subspecies, rather than botanical environment. Differentially abundant proteins were validated in sicula and ligustica honeys of different origin, by using the so-called "rectangular strategy", a proteomic approach commonly used for biomarker discovery in clinical proteomics. Then, machine learning was employed to identify which proteins were the most effective in distinguishing sicula and ligustica honeys. This strategy enabled the identification of two proteins, laccase-5 and venome serine protease 34 isoform X2, that were fully effective in predicting whether honey was made by sicula or ligustica honeybees. In conclusion, we profiled the proteome of sicula honey, identified two protein classifiers of sicula honey in respect to ligustica, and proved that the rectangular strategy can be applied to uncover biomarkers to ascertain food authenticity.

Multiple Bee Stings and Acute Kidney Injury: A Case Report.

Acute kidney injury (AKI) is a common complication following multiple honey bee stings and usually presents after 24-48 hours following the incidence. The severity of AKI is related to the number of stings. A single sting can cause an allergic reaction, and as the stings increase, a higher amount of venom is inoculated, leading to systemic poisoning. Bee venom can have direct or indirect effects on the kidneys. AKI is a combination of toxic and ischemic acute tubular necrosis. Patients may require dialysis, and the usual renal recovery time is 4-120 days. The patient with multiple honey bee stings needs emergency medical treatment, sometimes in the ICU setting, with the aim of treating or preventing anaphylaxis reactions. A case of AKI due to multiple honey bee stings is presented, which is rare but a known occurrence. The patient survived with a recovery of renal function.

Development of germline progenitors in larval queen honeybee ovaries.

Honeybees (Apis mellifera) are a keystone species for managed pollination and the production of hive products. Eusociality in honeybees leads to much of the reproduction in a hive driven by the queen. Queen bees have two large active ovaries that can produce large numbers of eggs if conditions are appropriate. These ovaries are also active throughout the long lives of these insects, up to 5 years in some cases. Recent studies have indicated that the germline precursors of the adult honeybee queen ovary are organized into 8-cell clusters, joined together by a polyfusome; a cytoplasmic bridge. To understand the origin of these clusters, and trace the development of the honeybee queen ovary, we examined the cell types and regionalization of the developing larval and pupal queen ovaries. We used established (nanos and castor), and novel (odd skipped) gene expression markers to determine regions of the developing ovary. Primordial germline cells develop in the honeybee embryo and are organized into ovary structures before the embryo hatches. The ovary is regionalized by larval stage 3 into terminal filaments and germaria. At this stage, clusters of germline cells in the germaria are joined by fusomes and are dividing synchronously. The origin of the 8-cell clusters in the adult germarium is therefore during larval stages. On emergence, the queen ovary has terminal filaments and germaria but has not yet developed any vitellaria, which are produced after the queen embarks on a nuptial flight. The lack of germaria, and the storing of germline progenitors as clusters, may be adaptions for queen bees to endure the metabolic demands of a nuptial flight, as well as rapidly lay large numbers of eggs to establish a hive.

Beekeeping breakthrough: unveiling hive health with a portable membrane inlet mass spectrometry detection method.

Supporting bee populations is essential considering threats posed by human activities like pesticide usage and habitat destruction. However, the current methods for monitoring and analyzing beehives and their surrounding environments are invasive, complex, and time-consuming. These methods often rely heavily on laboratory analyses, making them difficult to implement independently in the field. This study explores the application of portable membrane inlet mass spectrometer (MIMS) for noninvasive hive analysis, demonstrating its ability to detect various compounds indicative of hive conditions and environmental stressors. In addition to the expected compounds found in beehives, such as α-bergamotene, hexadecanoic acid, heptadecane, hexadecanamide, α-bisabolol-, 9-octadecenamide, (Z) - , and benzaldehyde, unexpected compounds, pollutants, like indane (polycyclic aromatic hydrocarbon) and carbofuran (pesticide), were also detected. The MIMS detection method provides rapid, accurate, and real-time results, making it suitable for preventive measures against bee diseases and integral to environmental biomonitoring. This integration of technology represents a significant advancement in bee conservation efforts, offering hope for the future of both bees and ecosystems.

Differences between uncapping and removal behaviors in Apis cerana from the perspective of long non-coding RNAs.

BACKGROUND: Hygienic behavior, a specialized form of immune response evolved in social insects, plays a crucial role in safeguarding colonies from disease spread. In honeybee colonies, such behavior typically entails the dual steps of uncapping and removal of unhealthy and deceased brood. Although in recent years, numerous studies have examined the development of hygienic behavior, the mechanisms underlying the division in the performance of uncapping and removal have yet to be sufficiently elucidated. In this regard, long non-coding RNAs (lncRNAs) have been evidenced to be engaged in regulating the physiological activities of honeybees; however, whether lncRNAs are likewise involved in the uncapping and removal tasks has not been clarified. RESULTS: In this study, the strong hygienic Apis cerana worker bees were used and the processes of uncapping and removal behaviors in three colonies were assayed with freeze-killed brood in the field. We then sequenced the antennal RNAs of honeybees to identify differentially expressed lncRNAs and performed lncRNA-mRNA association analysis to establish the differences between uncapping and removal. We detected 1,323 differentially expressed lncRNAs in the antennae, and the findings of lncRNA-mRNA association analyses revealed that the target genes of differentially expressed lncRNAs between uncapping and removal worker bees were predominantly linked to response to stimulus, receptor activity, and synapse. Notably, among the lncRNAs enriched in cellular response to stimulus, XR_001766094.2 was exclusively expressed in the uncapping worker bees. Based on these findings, we hypothesize that XR_001766094.2 plays a key role in distinguishing uncapping from removal behaviors by responding to external stimulus, thereby suggesting that the division of hygienic behaviors is governed by differential thresholds of responsiveness to environmental cues. CONCLUSION: We characterized differences in the uncapping and removal behaviors of worker bees from a perspective of lncRNAs. Uncapping bees may be equipped with a more rapid stimulatory response and more acute olfactory sensitivity, contributing to the rapid hygienic behavior in honeybee colonies. Our results thus establish a foundation for potential lncRNA-mediated gene expression regulation in hygienic behavior.

An orthology-based methodology as a complementary approach to retrieve evolutionarily conserved A-to-I RNA editing sites.

Adar-mediated adenosine-to-inosine (A-to-I) mRNA editing is a conserved mechanism that exerts diverse regulatory functions during the development, evolution, and adaptation of metazoans. The accurate detection of RNA editing sites helps us understand their biological significance. In this work, with an improved genome assembly of honeybee (Apis mellifera), we used a new orthology-based methodology to complement the traditional pipeline of (de novo) RNA editing detection. Compared to the outcome of traditional pipeline, we retrieved many novel editing sites in CDS that are deeply conserved between honeybee and other distantly related insects. The newly retrieved sites were missed by the traditional de novo identification due to the stringent criteria for controlling false-positive rate. Caste-specific editing sites are identified, including an Ile>Met auto-recoding site in Adar. This recoding was even conserved between honeybee and bumblebee, suggesting its putative regulatory role in shaping the phenotypic plasticity of eusocial Hymenoptera. In summary, we proposed a complementary approach to the traditional pipeline and retrieved several previously unnoticed CDS editing sites. From both technical and biological aspects, our works facilitate future researches on finding the functional editing sites and advance our understanding on the connection between RNA editing and the great phenotypic diversity of organisms.

Investigation of the biological activity and toxicity of bioactive silver nanoparticles synthesized via Vitex agnus-castus seed extract on honey bees.

In this study, silver nanoparticles (AgNPs) coated with bioactive molecules were synthesized via Vitex agnus-castus L. (VAC) seed extract (VAC/AgNPs). The synthesized VAC/AgNPs were characterized by surface plasmon resonance (SPR) ultraviolet-visible region spectroscopy (UV-Vis). The hydrodynamic size and surface charge analysis of the particles were measured with a Zeta sizer. The results of UV-Vis and Zeta analysis revealed that AgNPs were synthesized, the size distribution was nanoscale, and the solution was stable. The effects of the synthesized VAC/AgNPs and aqueous extract of VAC seeds on honeybees were investigated by means of lifespan test and histopathological analysis. It was determined that both VAC seed extract and VAC/AgNPs were non-toxic to honeybees at certain doses, positively affected their life span and contributed to their longevity in the life span test. Furthermore, no adverse effects were detected in terms of intestinal health in histopathological examinations. Therefore, VAC/AgNPs are considered to be a promising bioactive agent for honeybees.

An algae-based polymer material as a pesticide adjuvant for mitigating off-target drift.

Off-target pesticide drift from cropland is a major source of pesticide exposure to pollinating insects inhabiting crop and wildlands in the lower Mississippi Delta (LMD) in the USA. This study is aimed to develop a drift-reducing pesticide adjuvant that is less/nontoxic to honeybees. Ongoing toxicology experiments with two widely-used insecticides and sodium alginate (SA) pointed out reductions in honeybee mortality compared to an industry standard reference polyacrylamide (PAM). When used as an adjuvant to spray the same insecticides described above, SA did not interfere in killing the target pests. Therefore, SA has been tested as a drift-reducing pesticide adjuvant to protect honeybees. Spray experiments in the lab were carried out in four sets: (i) water only, (ii) water and adjuvant, (iii) water and pesticide, and (iv) water, pesticide and adjuvant. Each set contained 18 treatment combinations to cover the ranges in spray pressure (three), adjuvant dose (three), and spray nozzles (two). The droplet spectrum was analyzed using a P15 image analyzer. Diameters of 10 %, 50 % and 90 % volumes (DV10, DV50, and DV90), droplet velocity, standard deviation and relative span were measured. The drift reduction potential (DRP) of SA was analyzed by (i) dose, (ii) spray pressure, and (iii) nozzle type. The DRP of SA is compared to that of PAM. Additionally, three field experiments were carried out to analyze the efficiency of SA in reducing pesticide drift. The results from our experiments collectively indicate that SA has significant potential in mitigating drift as well as minimizing pesticide toxicity to honeybees.

Sensorizing a Beehive: A Study on Potential Embedded Solutions for Internal Contactless Monitoring of Bees Activity.

Winter is the season of main concern for beekeepers since the temperature, humidity, and potential infection from mites and other diseases may lead the colony to death. As a consequence, beekeepers perform invasive checks on the colonies, exposing them to further harm. This paper proposes a novel design of an instrumented beehive involving color cameras placed inside the beehive and at the bottom of it, paving the way for new frontiers in beehive monitoring. The overall acquisition system is described focusing on design choices towards an effective solution for internal, contactless, and stress-free beehive monitoring. To validate our approach, we conducted an experimental campaign in 2023 and analyzed the collected images with YOLOv8 to understand if the proposed solution can be useful for beekeepers and what kind of information can be derived from this kind of monitoring, including the presence of Varroa destructor mites inside the beehive. We experimentally found that the observation point inside the beehive is the most challenging due to the frequent movements of the bees and the difficulties related to obtaining in-focus images. However, from these images, it is possible to find Varroa destructor mites. On the other hand, the observation point at the bottom of the beehive showed great potential for understanding the overall activity of the colony.

From data to harvest: Leveraging ensemble machine learning for enhanced crop yield predictions across Canada amidst climate change.

Accurate crop yield predictions are crucial for farmers and policymakers. Despite the widespread use of ensemble machine learning (ML) models in computer science, their application in crop yield prediction remains relatively underexplored. This study, conducted in Canada, aims to assess the potential of five distinct ensemble ML models-Adaptive Boosting (AdaBoost), Gradient Boosting Machine (GBM), XGBoost, LightGBM, and Random Forest (RF)-in predicting crop yields chosen for their ability to manage complex datasets and their strong performance potential. The study integrated various factors, including climate variables, satellite-derived vegetation indices, soil characteristics, and honeybee census data. Data preparation comprised two main steps: first, climate variables were interpolated and averaged for croplands in ArcGIS Pro, along with averaging vegetation indices and soil characteristics. Honeybee census data was also incorporated. Second, the data was organized in Python to create a structured format for models' input. The models' accuracy was assessed using Root Mean Squared Error (RMSE), R-squared, and Mean Absolute Error (MAE). XGBoost emerged as the most accurate model, with the lowest MAE (68.70 for canola and 39.47 for soybeans), lowest RMSE (119.48 for canola and 102.39 for soybeans), and highest R-squared values (0.95 for canola and 0.96 for soybeans) on the test dataset. The study also assessed crop yields under various climate change scenarios, finding minimal variations across the scenarios, but significant negative impacts on canola and soybean yields across Canada. Honeybee colonies were identified as the most influential factor on crop yields, contributing 52.34 % to canola and 57.18 % to soybean yields. This research provides detailed crop yield maps of canola and soybeans at the Census Consolidated Subdivisions (CCS) level across Canada's agricultural landscape, offering valuable forecasts for localized decision-making. Additionally, it offers a proactive strategy for climate change preparedness, assisting farmers and stakeholders optimise resource allocation and manage risks effectively.

What can laboratory studies tell us about potential effects of pesticides on nontarget arthropods populations and communities in the field?

Over the past decades, concern has been increasing over reported declines in aboveground biodiversity on farmland. In many regions, data on the toxicity of pesticides to honeybees (Apis mellifera), but not wider nontarget arthropod (NTA) data, are required for pesticide registration. In Europe, the effects of pesticides on NTAs and honeybees have been the subject of regulatory risk assessment for more than 30 years, resulting in a large database. Although insecticides may be expected to affect NTA populations, solely identifying insecticidal modes of action for further NTA testing would result in redundancy among low-risk testing products and may also exclude other modes of action with potential effects in the field. This study assessed whether the honeybee acute risk assessment could provide any indication of the potential impact and recovery time of NTAs in cropped areas at the field scale and, if so, how it might be used in a tiered testing approach. The hazard quotients (HQs; foliar application rate/LR50) were derived for 151 active substances (32% insecticides, 28% fungicides, 38% herbicides, 2% plant growth regulators) for which toxicity data for established EU Tier 1 NTA indicator species (Typhlodromus pyri, Aphidius rhopalosiphi) and application rate data were available. These HQs were compared with published NTA HQ thresholds indicating the time to recovery of NTA populations and communities in field studies (>1 to >12 months). Using the same application rate data, honeybee acute risk quotient (RQ) and HQ were also determined and compared with NTA HQs and honeybee regulatory thresholds. These comparisons demonstrated that, where required, the current regulatory honeybee acute RQ of 0.4 or honeybee HQ of 50 can provide an efficient screening tier to target NTA testing at those products and uses with potential effects in the field where recovery may exceed 12 months. Integr Environ Assess Manag 2024;00:1-8. © 2024 SETAC.

Honeybee gut bacterial strain improved survival and gut microbiota homeostasis in Apis mellifera exposed in vivo to clothianidin.

Pesticides are causing honeybee mortality worldwide. Research carried out on honeybees indicates that application of pesticides has a significant impact on the core gut community, which ultimately leads to an increase in the growth of harmful pathogens. Disturbances caused by pesticides also affect the way bacterial members interact, which results in gut microbial dysbiosis. Administration of beneficial microbes has been previously demonstrated to be effective in treating or preventing disease in honeybees. The objective of this study was to measure under in vivo conditions the ability of two bacterial strains (the Enterobacter sp. and Pantoea sp.) isolated from honeybee gut to improve survival and mitigate gut microbiota dysbiosis in honeybees exposed to a sublethal clothianidin dose (0.1 ppb). Both gut bacterial strains were selected for their ability to degrade clothianidin in vitro regardless of their host-microbe interaction characteristics (e.g., beneficial, neutral, or harmful). To this end, we conducted cage trials on 4- to 6-day-old newly emerging honeybees. During microbial administration, we jointly monitored the taxonomic distribution and activity level of bacterial symbionts quantifying 16S rRNA transcripts. First, curative administration of the Pantoea sp. strain significantly improved the survival of clothianidin-exposed honeybees compared to sugar control bees (i.e., supplemented with sugar [1:1]). Second, curative administration of the Enterobacter sp. strain significantly mitigated the clothianidin-induced dysbiosis observed in the midgut structural network, but without improving survival. IMPORTANCE: The present work suggests that administration of bacterial strains isolated from honeybee gut may promote recovery of gut microbiota homeostasis after prolonged clothianidin exposure, while improving survival. This study highlights that gut bacterial strains hold promise for developing efficient microbial formulations to mitigate environmental pesticide exposure in honeybee colonies.

Lactobacillus Firm-5-derived succinate prevents honeybees from having diabetes-like symptoms.

The gut microbiome plays an important role in honeybee hormonal regulation and growth, but the underlying mechanisms are poorly understood. Here, we showed that the depletion of gut bacteria resulted in reduced expression of insulin-like peptide gene (ilp) in the head, accompanied by metabolic syndromes resembling those of Type 1 diabetes in humans: hyperglycemia, impaired lipid storage, and decreased metabolism. These symptoms were alleviated by gut bacterial inoculation. Gut metabolite profiling revealed that succinate, produced by Lactobacillus Firm-5, played deterministic roles in activating ilp gene expression and in regulating metabolism in honeybees. Notably, we demonstrated that succinate modulates host ilp gene expression through stimulating gut gluconeogenesis, a mechanism resembling that of humans. This study presents evidence for the role of gut metabolite in modulating host metabolism and contributes to the understanding of the interactions between gut microbiome and bee hosts.

Wing vein abnormality analysis in honeybee (Apis mellifera L. 1758) populations from Iran.

The insect wing is one of the most important characteristics that allowed insects to occupy most of the habitats on the planet. Honeybee wings has been the subject of studies on the venation abnormalities. A total of 424 honeybees from 14 locations were collected and all four wings were removed and examined for 19 abnormalities on the forewings and 6 abnormalities on the hindwings. In general, supernumerary veins were the most common abnormalities seen and abnormalities no. 23, 2, 6, 1, 5, 21, 10, 13 had the highest and abnormalities no. 11, 17, 18, 19, 20, and 25 had the lowest frequencies. All of the abnormalities had similar frequencies in the right and left wings in the population. In terms of correlation between 25 abnormalities, abnormality pairs AB3-AB13, AB6-AB7, AB7-AB8, AB10-AB12, AB16-AB17 on the forewing and AB2-AB23, AB12-AB20, AB12-AB24, AB13-AB21, AB16-AB25, and AB17-AB25 between the forewing and hindwing show significant positive correlations and abnormality pairs AB4-AB5, AB7-AB15 and AB8-AB9 on the forewing show significant negative correlations with each other. In terms of the differential occurrence of abnormalities , a few locations differed significantly from other locations. This study provides some insights into the nature of these abnormalities on the honeybee wings.

Review: Nutritional Needs of Honeybees and Legislation on Apiculture By-Products in Animal Nutrition.

Honeybees are some of the smallest farmed animals, and apiculture by-products, e.g., honey, beeswax, propolis, royal jelly, and pollen contribute to animal nutrition. For the effective production of these by-products, the optimal development and nutrient supply of the honeybee is required. Beginning with the development of the mouth and anal pores on the second day of embryonic development, the digestive tract differentiates into the mouth and fore-, mid-, and hindgut during the pupal stage. The various glands within the oral cavity are particularly important, secreting enzymes and substances that are crucial for digestion and hive nutrition, e.g., invertase and royal jelly. Honeybees rely on a specialized caste system, with worker bees collecting nectar, pollen, water, and resin for the nutrition of the entire hive. Macronutrients, including proteins, carbohydrates, and lipids, obtained primarily from pollen and nectar, are essential for the growth and development of larvae and the overall health of the colony. Inadequate nutrient intake can lead to detrimental effects on larval development, prompting cannibalism within the hive. Apiculture by-products possess unique nutritional and therapeutic properties, leading to a growing interest in the use of honey, beeswax, propolis, and pollen as a feed additive. In recent years, the use of apicultural by-products in animal nutrition has been primarily limited to in vivo studies, which have demonstrated various positive impacts on the performance of farm animals. Honey, beeswax, propolis, royal jelly, and pollen are listed feed stuffs according to Regulation (EC) No. 68/2013. However, for animal nutrition there is not any specific legal definition for these products and no legal requirements regarding their ingredients as given for honey or beeswax in European food law.