Chemical Composition and Nutritional Value of Royal Jelly Samples Obtained from Honey Bee (Apis mellifera) Hives Fed on Oak and Rapeseed Pollen Patties.

Young workers, i.e., nurse honey bees, synthesize and secrete royal jelly to feed the brood and queen. Since royal jelly is a protein-rich substance, the quality of royal jelly may be influenced by the consumption of feed with varying protein content. We tested whether honey bee (Apis mellifera) colonies compensates for the nutritional quality to produce royal jelly by feeding different pollen patties made of oak or rapeseed pollen. After harvesting royal jelly, we examined the chemical composition including proximate nutrients, amino acids, proteins, fatty acids, and minerals of royal jelly samples obtained from two treatments. The results revealed that pollen patties with different nutritional levels did not influence the nutritional composition except for the crude fat. The levels of 10-HDA, which serves as an indicator of the royal jelly quality, showed no significant difference between the oak and rapeseed treatments, with values of 1.9 and 2.1 g/100 g, respectively. However, we found some differences in the protein intensity, particularly the MRJP3 precursor, MRJP3-like, and glucose oxidase. This study suggests that honey bees may have mechanisms to compensate for nutritional standards to meet the brood's and queen's nutritional requirements during bee pollen collection, preserving bee bread and royal jelly secretion.

Phospholipase A2 activity is required for immune defense of European (Apis mellifera) and Asian (Apis cerana) honeybees against American foulbrood pathogen, Paenibacillus larvae.

Honeybees require an efficient immune system to defend against microbial pathogens. The American foulbrood pathogen, Paenibacillus larvae, is lethal to honeybees and one of the main causes of colony collapse. This study investigated the immune responses of Apis mellifera and Apis cerana honeybees against the bacterial pathogen P. larvae. Both species of honeybee larvae exhibited significant mortality even at 102 103 cfu/mL of P. larvae by diet-feeding, although A. mellifera appeared to be more tolerant to the bacterial pathogen than A. cerana. Upon bacterial infection, the two honeybee species expressed both cellular and humoral immune responses. Hemocytes of both species exhibited characteristic spreading behaviors, accompanied by cytoskeletal extension along with F-actin growth, and formed nodules. Larvae of both species also expressed an antimicrobial peptide called apolipophorin III (ApoLpIII) in response to bacterial infection. However, these immune responses were significantly suppressed by a specific inhibitor to phospholipase A2 (PLA2). Each honeybee genome encodes four PLA2 genes (PLA2A ~ PLA2D), representing four orthologous combinations between the two species. In response to P. larvae infection, both species significantly up-regulated PLA2 enzyme activities and the expression of all four PLA2 genes. To determine the roles of the four PLA2s in the immune responses, RNA interference (RNAi) was performed by injecting gene-specific double stranded RNAs (dsRNAs). All four RNAi treatments significantly suppressed the immune responses, and specific inhibition of the two secretory PLA2s (PLA2A and PLA2B) potently suppressed nodule formation and ApoLpIII expression. These results demonstrate the cellular and humoral immune responses of A. mellifera and A. cerana against P. larvae. This study suggests that eicosanoids play a crucial role in mediating common immune responses in two closely related honeybees.

Uncovering floral composition of paper wasp nests (Hymenoptera: Vespidae: Polistes) through DNA metabarcoding.

As the social organism, Polistes wasps build a communal nest using woody fibers with saliva for sustaining brood and adult population throughout the season. Limited information exists regarding the identification specific plant materials employed in wasp nest building. Thus, we firstly tested if the DNA metabarcoding approach utilizing rbcL and trnL molecular markers could identify the plant species quantitatively and qualitatively inform the mixed-origin woody samples. A threshold of 0.01 proportion of reads was applied for rbcL and trnL molecular markers, while this threshold for median proportion was 0.0025. In assessing taxa richness, the median proportion demonstrated superior performance, exhibiting higher taxa detection power, however, rbcL marker outperformed in quantitative analysis. Subsequently, we applied DNA metabarcoding to identify the plant materials from the nests of two Polistes species, P. mandarinus and P. rothneyi. The results showed that higher preference of Quercus and Robinia as the major nest building materials regardless of the surrounding plant communities, by two wasp species. Material diversity was higher for P. rothneyi than P. mandarinus, which may explain the abundance of this species possibly with heightened adaptive capacities in their nesting behavior. This study demonstrated that DNA metabarcoding could identify the complex nest-building plant materials of paper wasps and provide insights into their ecological interactions in the natural ecosystem.

Global Trends in Climate Suitability of Bees: Ups and Downs in a Warming World.

Bees represent vital natural assets contributing significantly to global food production and the maintenance of ecosystems. While studies on climate change effects impacting major pollinators like honeybees and bumblebees raise concerns about global diversity and crop productivity, comprehensive global-scale analyses remain limited. This study explores the repercussions of global warming on 1365 bees across seven families of bees worldwide. To compile a robust global bee occurrence dataset, we utilized the innovative 'BeeBDC' R package that amalgamated over 18.3 million bee occurrence records sourced from various repositories. Through species distribution models under the SSP585 scenario in the year 2070, we assessed how climate change influences the climate suitability of bees on a global scale, examining the impacts across continents. Our findings suggested that approximately 65% of bees are likely to witness a decrease in their distribution, with reductions averaging between 28% in Australia and 56% in Europe. Moreover, our analysis indicated that climate change's impact on bees is projected to be more severe in Africa and Europe, while North America is expected to witness a higher number (336) of bees expanding their distribution. Climate change's anticipated effects on bee distributions could potentially disrupt existing pollinator-plant networks, posing ecological challenges that emphasize the importance of pollinator diversity, synchrony between plants and bees, and the necessity for focused conservation efforts.

Exploring the Functional Properties of Propolis, Geopropolis, and Cerumen, with a Special Emphasis on Their Antimicrobial Effects.

Bee propolis has been touted as a natural antimicrobial agent with the potential to replace antibiotics. Numerous reports and reviews have highlighted the functionalities and applications of the natural compound. Despite much clamor for the downstream application of propolis, there remain many grounds to cover, especially in the upstream production, and factors affecting the quality of the propolis. Moreover, geopropolis and cerumen, akin to propolis, hold promise for diverse human applications, yet their benefits and intricate manufacturing processes remain subjects of intensive research. Specialized cement bees are pivotal in gathering and transporting plant resins from suitable sources to their nests. Contrary to common belief, these resins are directly applied within the hive, smoothed out by cement bees, and blended with beeswax and trace components to create raw propolis. Beekeepers subsequently harvest and perform the extraction of the raw propolis to form the final propolis extract that is sold on the market. As a result of the production process, intrinsic and extrinsic factors, such as botanical origins, bee species, and the extraction process, have a direct impact on the quality of the final propolis extract. Towards the end of this paper, a section is dedicated to highlighting the antimicrobial potency of propolis extract.

Metabolomic analysis reveals linkage between chemical composition and sensory quality of different floral honey samples.

Honey has a distinct flavor characterized by various volatiles and non-volatiles from diverse origins. In this study, metabolomics combined with sensory analysis was performed to identify relationships between chemical profile and sensory quality of honey. Targeted metabolomic analysis was conducted to determine volatile and non-volatile profiles of seven different honey. Volatile profile was analyzed using headspace solid-phase microextraction (HS-SPME) coupled to GC - MS. LC - MS/MS, HPLC - UV, and HPLC-RI were employed to analyze flavonoids, organic acids, and sugars, respectively. Authentic standards were utilized for confirmation of metabolites. Sensory evaluation included quantitative descriptive analysis and consumer acceptance test. The results showed that sucrose (sweetness) was responsible for a positive hedonic perception, while organic acids and flavonoids (sourness, astringency, bitterness) negatively affected consumer acceptance. Volatiles with floral notes (e.g. decyl formate) were preferred, but others with off-flavors (e.g. 2-methylbenzofuran) were not preferred by consumers. Flavor familiarity was strongly correlated with the consumer acceptance of honey, indicating that the balance between volatiles and non-volatiles is significant for honey flavor quality. This work demonstrates the role of key flavor compounds in honey quality, and may be applicable to the quality control of honey.

The first record of the genus Julolaelaps Berlese (Acari, Mesostigmata, Laelapidae) in Republic of Korea and description of a new species on a captive giant African millipede (Spirostreptidae, Archispirostreptus).

This paper reports on a new species of mite of the genus Julolaelaps Berlese in Republic of Korea. Females and males of a new species, Julolaelapsgigassp. nov., were collected on a captive giant African millipede, Archispirostreptusgigas (Spirostreptida: Spirostreptidae). The new species is described and illustrated based on morphological characters of the adult females and males and compared with known congeners. This new species is the first record of Julolaelaps from Republic of Korea. In addition, an updated key to all known species of the genus is presented.

Acaricidal Toxicity of Four Essential Oils, Their Predominant Constituents, Their Mixtures against Varroa Mite, and Their Selectivity to Honey Bees (Apis cerana and A. mellifera).

The honey bee (Apis mellifera) faces a significant threat from Varroa destructor, causing the losses of millions of colonies worldwide. While synthetic acaricides are widely used to control Varroa infestations, excessive application has led to resistant strains and poses side effects on the host. Consequently, there is an urgent need for a new acaricide that is both effective and affordable, yet safe to use on bees. One potential source of these acaricides is essential oils (EOs) and their constituents. This study evaluated the acaricidal properties of four essential oils (Eucalyptus globulus, Rosemary officinalis, Trachyspermum ammi (Ethiopian and Indian varieties), their constituents and mixture of constituents against V. destructor through the complete exposure method. Our finding showed that a 1:1 mixture of thymol and carvacrol (4 h-LC50 = 42 µg/mL), thymol (4 h-LC50 = 71 µg/mL), and T. ammi oil (4 h-LC50 = 81-98 µg/mL) were the most toxic test samples against V. destructor. Honey bee behavior and selectivity were also assessed with one additional EO Thymus schimperi, indicating that T. schimperi, T. ammi, and their components were selective and did not affect the learning and memory of bees. In conclusion, the thymol and carvacrol (1:1) mixture was shown to be a promising replacement for synthetic acaricides, being three times more toxic than a commercial acaricide, fluvalinate (4 h-LC50 = 143 µg/mL).

Modeling abundance and risk impact of Vespa velutina nigrithorax (Hymenoptera: Vespidae) in Korea: application of a species abundance model.

The Asian yellow-legged hornet, Vespa velutina nigrithorax, is native to Southeast Asia. However, it has invaded many countries in temperate regions, causing serious threats to honeybees and human health. With a growing demand for estimating the potential distribution of this species, multiple studies have resorted to occurrence-based models. However, they are less informative for predicting local abundance patterns associated with the species' impact. Thus, we aimed to develop an abundance-based distribution model for V. v. nigrithorax in Korea to support the forecast of its impact and associated management strategies. The abundance data of V. v. nigrithorax were collected from 254 sites for 4 years covering the country and used to develop a model with bioclimatic and land composition variables. Along with the abundance model, the classical occurrence model was tested to determine whether it could provide a reasonable prediction on the estimation of local abundance. As a result, the abundance model provided higher discriminative power and accuracy than the occurrence model to evaluate the impacts caused by V. v. nigrithorax. On the other hand, the occurrence model was not able to discriminate abundance in the areas occupied by V. v. nigrithorax, indicating an unclear occurrence-abundance relationship or oversimplification of the estimated niche created by the occurrence model. Based on the final abundance model, risk indices for human health and honeybee losses were suggested. These results could help to provide support for risk management of V. v. nigrithorax in Korea and to give biological information to other countries where this species has already become established or which it is likely to invade in the near future.

Honey Quality Control: Review of Methodologies for Determining Entomological Origin.

Honey is a widely consumed natural product, and its entomological origin can significantly influence its market value. Therefore, traceability of the entomological origin of honey should also be considered in honey quality control protocols. Although several methods exist, such as physicochemical characterization and bioactivity profiling of honey of different entomological origins, the most promising three methods for entomological authentication of honey include protein-based identification, chemical profiling, and a DNA-based method. All of these methods can be applied for reliable identification of the entomological origin of honey. However, as the honey is a complex matrix, the inconsistency of the results obtained by these methods is a pragmatic challenge, and therefore, the use of each method in all the cases is questionable. Most of these methodologies can be used for authentication of newly harvested honey and it is worth understanding the possibility of using these methods for authentication of relatively old samples. Most probably, using DNA-based methods targeting small fragments of DNA can provide the best result in old samples, however, the species-specific primers targeting short fragments are limited and not available for all species. Therefore, using universal primers in combination with a DNA metabarcoding approach can be a good solution that requires further investigation. This present article describes the applications of different methods, their pros, and their cons to identify honey based on entomological origin.

Metagenomic analysis of viromes in honey bee colonies (Apis mellifera; Hymenoptera: Apidae) after mass disappearance in Korea.

After the nationwide, massive winter losses of honey bees in Korea during the winter of 2021, samplings were conducted from live honey bees in colonies and dead honey bees nearby colonies in the same bee-farms in six regions in Korea. Each sample was subjected to virome analysis using high-throughput sequencing technology. The number of viral reads was the lowest in the live honey bee group sample with 370,503 reads and the highest in the dead honey bee group sample with 42,659,622 reads. Viral contigs were matched with the viral genomes of the black queen cell virus, deformed wing virus, Israeli acute paralysis virus, and sacbrood virus, all of which have been previously reported in Korea. However, Apis rhabdovirus 5, bee macula-like virus, Varroa orthomyxovirus-1, Hubei partiti-like virus 34, Lake Sinai virus 2, 3, and 4, and the Ditton virus, were also discovered in this study, which are the first records in Korea. Plant viral sequences resembling those of Arabidopsis latent virus 1, and a novel viral sequence was also discovered. In the present study 55 complete viral genome sequences were identified. This study is the first virome analysis of domestic honey bees and provides the latest information on the diversity of honey bee viruses in Korea.

The Effect of Apple-Derived Nanovesicles on the Osteoblastogenesis of Osteoblastic MC3T3-E1 Cells.

Osteoporosis is characterized by low bone mass and elevated structural deterioration of the bone tissue, resulting in bone weakness with an increased risk of fracture. Considering biological activities of various phytochemicals extracted from apples, we herein demonstrated the potential antiosteoporotic effects of apple-derived nanovesicles (apple NVs) using osteoblastic MC3T3-E1 cells. Apple NVs significantly stimulated the growth of MC3T3-E1 cells. The cellular alkaline phosphatase (ALP) activity was significantly upregulated in the 5 µg/mL apple NVs-treated group. In addition, the concentrarion of mineralized nodules was significantly increased in the apple NVs-treated groups. Furthermore, apple NVs increased the expression of the genes and proteins associated with osteoblast growth and differentiation, such as Runx2, ALP, OPN, and BMP2/4, which further activated ERK- and JNK-related mitogen-activated protein kinase signaling. These results demonstrate that apple NVs have a potential to prevent osteoporosis by promoting osteoblastogenesis in osteoblastic MC3T3-E1 cells through regulating the BMP2/Smad1 pathways.

Embracing Tradition: The Vital Role of Traditional Foods in Achieving Nutrition Security.

In a press release from the Convention on Biological Diversity (CBD) in 2019, it was reported that, currently, a mere 12 plants and 5 animal species account for approximately 75% of global food production [...].

Differential Bacterial Community of Bee Bread and Bee Pollen Revealed by 16s rRNA High-Throughput Sequencing.

We investigated the bacterial community of bee bread and bee pollen samples using an approach through 16 s rRNA high-throughput sequencing. The results revealed a higher bacterial diversity in bee bread than in bee pollen as depicted in taxonomic profiling, as well as diversity indices such as the Shannon diversity index (3.7 to 4.8 for bee bread and 1.1 to 1.7 for bee pollen samples) and Simpson's index (>0.9 for bee bread and 0.4−0.5 for bee pollen). Principal component analysis showed a distinct difference in bacterial communities. The higher bacterial diversity in the bee bread than bee pollen could presumably be due to factors such as storage period, processing of food, fermentation, and high sugar environment. However, no effect of the feed (rapeseed or oak pollen patties or even natural inflow) was indicated on the bacterial composition of bee bread, presumably because of the lack of restriction of foraged pollen inflow in the hive. The diverse bacterial profile of the bee bread could contribute to the nutritional provisioning as well as enhance the detoxification process; however, a thorough investigation of the functional role of individual bacteria genera remains a task for future studies.

Honey DNA metabarcoding revealed foraging resource partitioning between Korean native and introduced honey bees (Hymenoptera: Apidae).

Honey DNA metabarcoding provides information of floral sources of honey and foraging plant preferences of honey bees. We evaluated the floral composition of honey from two different species of honey bees, Apis cerana honey (ACH) and A. mellifera honey (AMH) in a mixed apiary located in a semi-forest environment to understand the floral preference and level of interspecific competition on floral resource. Three honey samples were collected from different hives of each species in mid-August. In total, 56 plant taxa were identified across the honey samples and among them, 38 taxonomic units were found in ACH compared with a total of 33 in AMH. The number of major plants (> 1% of reads) in honey samples was 9 and 11 in ACH and AMH respectively indicating the higher diversity of plant taxa in AMH. 23 taxonomic units were found exclusively in ACH, 18 taxonomic units were found only in AMH and 15 taxonomic units were shared between ACH and AMH indicating that 73% of the taxonomic units were present only in honey originated from one of the honeybee species. Qualitative and quantitative analyses of the shared major plants revealed the division of floral resource between these co-existing honey bee species pointing to a low level of interspecific competition between these two important pollinators.

DNA-Based Method for Traceability and Authentication of Apis cerana and A. dorsata Honey (Hymenoptera: Apidae), Using the NADH dehydrogenase 2 Gene.

Honey is a widely used natural product and the price of honey from Apis cerana (ACH) and A. dorsata (ADH) is several times more expensive than the one from A. mellifera (AMH), thus there are increasing fraud issues reported in the market by mislabeling or mixing honeys with different entomological origins. In this study, three species-specific primers, targeting the NADH dehydrogenase 2 (ND2) region of honeybee mitochondrial DNA, were designed and tested to distinguish the entomological origin of ACH, ADH, and AMH. Molecular analysis showed that each primer set can specifically detect the ND2 region from the targeted honeybee DNA, but not from the others. The amplicon size for A. cerana, A. dorsata and A. mellifera were 224, 302, and 377 bp, respectively. Importantly, each primer set also specifically produced amplicons with expected size from the DNA prepared from honey samples with different entomological origins. The PCR adulteration test allowed detection of 1% of AMH in the mixture with either ACH or ADH. Furthermore, real-time PCR and melting curve analysis indicated the possible discrimination of origin of honey samples. Therefore, we provide the newly developed PCR-based method that can be used to determine the entomological origin of the three kinds of honey.

Interest in Insects as Food and Feed: It Does Not Wane in the Public Domain.

This Special Issue of Foods represents Volume 2 of the topic "Edible Insects as Innovative Foods: Nutritional, Functional and Acceptability Assessments" [...].

Nutritional Composition of Honey Bee Drones of Two Subspecies Relative to Their Pupal Developmental Stages.

We examined the contents of nutritional importance, i.e., amino acids, fatty acids and minerals of different developmental stages of drones of two honey bee subspecies, namely Apis mellifera carnica and A. m. mellifera. The results revealed that, in general, individual amino acid amounts and therefore the total protein increased along with the developmental stages of the drones. No statistically significant differences were found between the same developmental stages of the two subspecies. The reverse, i.e., a decrease with developmental stage occurred in relation to the fatty acid composition. Most of the minerals were higher at advanced developmental stages. Overall, the high protein content (31.4-43.4%), small amount of fat (9.5-11.5%) and abundance of minerals such asiron and zinc, make drones a suitable nutritional resource. Even though nutrient content, especially protein, was higher in the pupae than the prepupae, we propose prepupae also as a commercial product based on their higher biomass production. Provided standard production protocols maintaining hygiene and safety will be adhered to, we propose that drone honey bees can be utilized as human food or animal feed.

Lactucin induces apoptosis through reactive oxygen species-mediated BCL-2 and CFLARL downregulation in Caki-1 cells.

BACKGROUND: Lactucin, a naturally occurring active sesquiterpene lactone, is abundantly found in chicory and romaine lettuce. A recent study reported that lactucin could induce apoptosis in leukemia cells. However, its cytotoxicity and potential molecular mechanisms underlying cancer cell death remain unclear. OBJECTIVE: Therefore, in this study, we aimed to investigate the direct effect and underlying mechanism of action of lactucin on renal cancer cells. METHODS: MTT assay and flow cytometry were performed to evaluate the rate of cell proliferation and apoptosis, respectively. Western blotting, reverse transcription polymerase chain reaction, and protein stability analyses were performed to analyze the effect of lactucin on the expression of apoptosis-related proteins such as B-cell lymphoma 2 (BCL-2) and CFLAR (CASP8 and FADD like apoptosis regulator) long isoform (CFLARL) in Caki-1 human renal cancer cells. In addition, reactive oxygen species (ROS) generation was evaluated using flow cytometry. RESULTS: Lactucin treatment induced apoptosis in Caki-1 cells in a dose-dependent manner via activation of the caspase pathway. It downregulated BCL-2 and CFLARL expression levels by suppressing BCL-2 transcription and CFLARL protein stability, respectively. Pretreatment with N-acetyl-1-cysteine, a ROS scavenger, attenuated the lactucin-induced apoptosis and restored the BCL-2 and CFLARL expression to basal levels. Lactucin-facilitated BCL-2 downregulation was regulated at the transcriptional level through the inactivation of the NF-κB pathway. CONCLUSIONS: Our study is the first to demonstrate that lactucin-induced apoptosis is mediated by ROS production, which in turn activates the caspase-dependent apoptotic pathway by inhibiting BCL-2 and CFLARL expression in Caki-1 cells.

Chemical Composition, Nutrient Quality and Acceptability of Edible Insects Are Affected by Species, Developmental Stage, Gender, Diet, and Processing Method.

Edible insects have been considered as either nutritious food itemsper se, or as wholesome ingredients to various dishes and components of traditional subsistence. Protein, fat, mineral and vitamin contents in insects generally satisfy the requirements of healthy food, although there is considerable variation associated with insect species, collection site, processing method, insect life stage, rearing technology and insect feed. A comparison of available data(based on dry weight) showed that processing can improve the nutrient content, taste, flavour, appearance and palatability of insects, but that there are additional factors, which can impact the content and composition of insect species that have been recommended for consumption by humans. This review focuses on factors that have received little attention in connection with the task to improve acceptability or choice of edible insects and suggests ways to guarantee food security in countries where deficiencies in protein and minerals are an acute and perpetual problem. This review is meant to assist the food industry to select the most suitable species as well as processing methods for insect-based food products.