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The “dark honeybee” Apis mellifera mellifera, originally the dominating subspecies of the Western honeybee in Central and Northern Europe, is currently threatened by extinction in Austria and its conservation breeding program is carried out by two breeding associations in Salzburg (AMZ – Austrian Mellifera Breeders) and Tyrol (breeding association Kauns-Kaunerberg). In this study, genetic diversity (genomic inbreeding FROH, number of homozygosity segments NROH), the relationship structures and foreign gene proportions in samples from the two Mellifera breeding associations were examined in comparison to A.m.carnica samples using the high-density 70k SNP Illumina® Honey Bee Genotyping Array. The mean Carnica gene-proportion (C-line gene proportion) in the two Mellifera samples was originally 32.4 to 34.4%. By excluding hybridized queens, this gene share was reduced to an average of 12.8–17.7%. In terms of genomic inbreeding, the Austrian Mellifera samples exhibited a mean FROH of 4.20% with maximum values of up to 17.2%. The results of this study clearly illustrated the problems conservation breeding programs in small populations are exposed to. We further demonstrated that the simultaneous consideration of multiple genetic descriptors such as FROH, NROH, ADMIXTURE and the kinship network NETVIEW is effective and practically usable in monitoring breeding programs, which can further be communicated in a way that is understandable to the practical breeders. [ABSTRACT FROM AUTHOR]

Background: Neonicotinoids are widespread insecticides because of their potent effects against aphids and other piercing-sucking insects in addition to having high selectivity toward insects rather than vertebrates. However, they affect severely some non-target insects, mainly honeybee in a phenomenon called colony collapse disorder (CCD). Results: Effects of imidacloprid (IMI), most used neonicotinoids, on aphid acetylcholinesterase (AChE), in vivo and in vitro were examined; besides, molecular modeling was used to investigate similarities and differences of AChE and nicotinic acetylcholine receptors α1-subunit (nAChRα1) in aphids, target insect, and honeybees, non-target insect. Results showed that aphid AChE was inhibited in vitro, with IC50 108.6 mg/L but not affected in vivo while the mortality was concentration-dependent with high toxicity (LC50 9.50 mg/L); in addition, aphid AChE was more inhibited, in vitro, but with much less effects, in vivo, than that of honeybees. These results indicate that AChE is not the main cause of the observed mortality, but it still has a role in insect resistance system with different responses in both insects. Molecular modeling showed high similarity in primary and secondary structures of AChE indicated by high identity (67%) and low gaps (1%); besides, the same template for both enzymes was auto-selected for homology. In addition, similar positions of the triad amino acids were found in AChE of both insects indicating high similarity. Conversely, the similarity in nAChRα1 in both insects is lower (50% identity and 9% gaps). These gaps (50 amino acids) are found in the intracellular large loop between TM3 and TM4 and account for the observed differences in the nAChRα1 binding sites of in both insects. Conclusion: These observed variations in nAChRα1 structures and binding sites in different insect species can be used as good bases in designing new neonicotinoids that express high effects on target insects with better selectivity to minimize adverse effects on non-target organisms. [ABSTRACT FROM AUTHOR]

Despite the major role that insect pollinators play in crop production, agricultural intensification drives them into decline. Various conservation measures have been developed to mitigate the negative effects of agriculture on insect pollinators. In a novel comparison of the efficacy of three conservation measures on honeybee colony growth, we monitored experimental honeybee colonies in 16 landscapes that comprised orthogonal gradients of organic agriculture, annual flower strips and perennial semi‐natural habitats. Using structural equation modelling, we assessed the effects of conservation measures on the prevalence of 11 parasites, Varroa destructor loads and their collective impact on colony growth. Increasing area coverage of perennial semi‐natural habitat related to higher V. destructor load and indirectly to lower colony growth. Increasing area of annual flower strips was associated with lower V. destructor load and indirectly with higher colony growth. Increasing area of organic farming related to lower parasite richness and also directly to improved colony growth. Synthesis and applications: Landscape features can affect pollinators directly through the provision of food resources and indirectly through modulation of parasite prevalence. To promote honeybee colony health in agro‐ecosystems, our results suggest that organic agriculture and annual flower strips should be prioritized conservation measures. Landscape management should consider the merits and demerits of different measures to sustain healthy populations of pollinators in agro‐ecosystems. [ABSTRACT FROM AUTHOR]

Our understanding of the nervous tissues that affect the wing flapping of insects mainly focuses on the brain, but wing flapping is a rhythmic movement related to the central pattern generator in the ventral nerve cord. To verify whether the neural activity of the abdominal ganglion of the honeybee (Apis mellifera) affects the flapping‐wing flight, we profiled the response characteristics of indirect flight muscles to abdominal ganglion excitation. Strikingly, a change in the neural activity of ganglion 3 or ganglion 4 has a stronger effect on the electrophysiological activity of indirect flight muscles than that of ganglion 5. The electrophysiological activity of vertical indirect flight muscles is affected more by the change in neural activity of the abdominal ganglion than that of lateral indirect flight muscles. Moreover, the change in neural activity of the abdominal ganglion mainly causes the change in the muscular activity of indirect wing muscles, but the activity patterns change relatively little and there is little change in the complicated details. This work improves our understanding of the neuroregulatory mechanisms associated with the flapping‐wing flight of honeybees. [ABSTRACT FROM AUTHOR]

The queen bee is the core member of a bee colony, and her loss will pose a great threat to the survival of the colony that may cause colony collapse. However, the process by which queen bee loss affects the internal social state of the bee colony remains unclear. In this paper, we used a multi-sensors system to continually monitor colonies with queen loss and regularly checked their biological status. Our results show that the queen loss initially caused a rapid decrease in brood rearing and changed the foraging strategy of the colony, leading to an increase in food storage. Also the population decline is difficult to reverse in a short time, even if the queen is naturally replaced. This study emphasises the impact of queen bee loss on the operation of the bee colony social system, and elucidates the interconnectedness of the bee colony social system. • Bee colony status was monitored using biology and multiple sensors. • Queen bee loss alters internal states as brooding, foraging, hive weight. • A new-born queen bee cannot improve the internal colony state in the short term. • A single indicator change can trigger chain reaction in the bee colony system. • Monitoring hive weight changes can effectively identify queen bee loss. [ABSTRACT FROM AUTHOR]

Simple Summary: Interest in beekeeping has significantly increased in recent years. A basic understanding of the nutritional and physiological requirements of honeybees is of great interest to improve their welfare and honey yields. The unique anatomic features of the gastrointestinal tract of bees, which enable bees to make optimal use of their food resources, are extensively described in this review. Nectar, pollen, and resins are raw materials for the synthesis of honey, beeswax, propolis, and royal jelly by the honeybee. All of these products are used for nutrition or as food supplements for animals. In the context of these products, the legal requirements for the use of apiculture by-products in animal nutrition are highlighted. 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. [ABSTRACT FROM AUTHOR]

Bee venom serves as an essential defensive weapon for bees and also finds application as a medicinal drug. MicroRNAs (miRNAs) serve as critical regulators and have been demonstrated to perform a variety of biological functions. However, the presence of miRNAs in bee venom needs to be confirmed. Therefore, we conducted small RNA sequencing and identified 158 known miRNAs, 15 conserved miRNAs and 4 novel miRNAs. It is noteworthy that ame‐miR‐1‐3p, the most abundant among them, accounted for over a quarter of all miRNA reads. To validate the function of ame‐miR‐1‐3p, we screened 28 candidate target genes using transcriptome sequencing and three target gene prediction software (miRanda, PITA and TargetScan) for ame‐miR‐1‐3p. Subsequently, we employed real‐time quantitative reverse transcription PCR (qRT‐PCR), Western blot and other technologies to confirm that ame‐miR‐1‐3p inhibits the relative expression of antizyme inhibitor 1 (AZIN1) by targeting the 3′ untranslated region (UTR) of AZIN1. This, in turn, caused ODC antizyme 1 (OAZ1) to bind to ornithine decarboxylase 1 (ODC1) and mark ODC1 for proteolytic destruction. The reduction in functional ODC1 ultimately resulted in a decrease in polyamine biosynthesis. Furthermore, we determined that ame‐miR‐1‐3p accelerates cell death through the AZIN1/OAZ1‐ODC1‐polyamines pathway. Our studies demonstrate that ame‐miR‐1‐3p diminishes cell viability and it may collaborate with sPLA2 to enhance the defence capabilities of honeybees (Apis mellifera L.). Collectively, these data further elucidate the defence mechanism of bee venom and expand the potential applications of bee venom in medical treatment. [ABSTRACT FROM AUTHOR]

Human activities associated with large-scale farms and the monocultures expose honey bees to one type of food. Moreover, there is an ongoing decline of plant species producing pollen and nectar in Europe. A poorly balanced diet affects a number of processes occurring in a bee's body. The fat body and hemolymph are the tissues that participate in all of them. Therefore, the aim of our study was to determine the effect of hazel, pine, rapeseed, buckwheat, phacelia and goldenrod pollen on the morphological parameters of fat body trophocytes, the diameters of cell nuclei in oenocytes and the concentrations of compounds involved in energy metabolism (glucose, glycogen, triglycerides and protein). In the cage tests, the bees were fed from the first day of life with sugar candy (control group) or candy with a 10% addition of one of the 6 pollen types. Hemolymph and fat body from various locations were collected from 1-, 7- and 14-day-old workers. Pollen produced by plant species such as hazel and pine increased glucose concentrations in the bee tissues, especially in the hemolymph. It can therefore be concluded that they are valuable sources of energy (in the form of simple carbohydrates) which are quickly used by bees. Pollen from plants blooming in the summer and autumn increased the concentrations of proteins, glycogen and triglycerides in the fat body, especially that from the third tergite. The accumulation of these compounds was associated with an increased the length and width of trophocytes as well as with enhanced metabolic activity, which was evidenced in the increasing diameter of oenocyte cell nuclei. It seems a balanced multi-pollen diet is more valuable for bees, but it is important to understand the effects of the particular pollen types in the context of a mono-diet. In the future, this will make it possible to produce mixtures that can ensure homeostasis in the apian body. [ABSTRACT FROM AUTHOR]

Abstract Background Neonicotinoids are widespread insecticides because of their potent effects against aphids and other piercing-sucking insects in addition to having high selectivity toward insects rather than vertebrates. However, they affect severely some non-target insects, mainly honeybee in a phenomenon called colony collapse disorder (CCD). Results Effects of imidacloprid (IMI), most used neonicotinoids, on aphid acetylcholinesterase (AChE), in vivo and in vitro were examined; besides, molecular modeling was used to investigate similarities and differences of AChE and nicotinic acetylcholine receptors α1-subunit (nAChRα1) in aphids, target insect, and honeybees, non-target insect. Results showed that aphid AChE was inhibited in vitro, with IC50 108.6 mg/L but not affected in vivo while the mortality was concentration-dependent with high toxicity (LC50 9.50 mg/L); in addition, aphid AChE was more inhibited, in vitro, but with much less effects, in vivo, than that of honeybees. These results indicate that AChE is not the main cause of the observed mortality, but it still has a role in insect resistance system with different responses in both insects. Molecular modeling showed high similarity in primary and secondary structures of AChE indicated by high identity (67%) and low gaps (1%); besides, the same template for both enzymes was auto-selected for homology. In addition, similar positions of the triad amino acids were found in AChE of both insects indicating high similarity. Conversely, the similarity in nAChRα1 in both insects is lower (50% identity and 9% gaps). These gaps (50 amino acids) are found in the intracellular large loop between TM3 and TM4 and account for the observed differences in the nAChRα1 binding sites of in both insects. Conclusion These observed variations in nAChRα1 structures and binding sites in different insect species can be used as good bases in designing new neonicotinoids that express high effects on target insects with better selectivity to minimize adverse effects on non-target organisms. Graphical Abstract

Abstract 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.

Abstract Human activities associated with large-scale farms and the monocultures expose honey bees to one type of food. Moreover, there is an ongoing decline of plant species producing pollen and nectar in Europe. A poorly balanced diet affects a number of processes occurring in a bee’s body. The fat body and hemolymph are the tissues that participate in all of them. Therefore, the aim of our study was to determine the effect of hazel, pine, rapeseed, buckwheat, phacelia and goldenrod pollen on the morphological parameters of fat body trophocytes, the diameters of cell nuclei in oenocytes and the concentrations of compounds involved in energy metabolism (glucose, glycogen, triglycerides and protein). In the cage tests, the bees were fed from the first day of life with sugar candy (control group) or candy with a 10% addition of one of the 6 pollen types. Hemolymph and fat body from various locations were collected from 1-, 7- and 14-day-old workers. Pollen produced by plant species such as hazel and pine increased glucose concentrations in the bee tissues, especially in the hemolymph. It can therefore be concluded that they are valuable sources of energy (in the form of simple carbohydrates) which are quickly used by bees. Pollen from plants blooming in the summer and autumn increased the concentrations of proteins, glycogen and triglycerides in the fat body, especially that from the third tergite. The accumulation of these compounds was associated with an increased the length and width of trophocytes as well as with enhanced metabolic activity, which was evidenced in the increasing diameter of oenocyte cell nuclei. It seems a balanced multi-pollen diet is more valuable for bees, but it is important to understand the effects of the particular pollen types in the context of a mono-diet. In the future, this will make it possible to produce mixtures that can ensure homeostasis in the apian body.

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.

Nosema ceranae, a parasite that parasitizes and reproduces in the gut of honeybees, has become a serious threat to the global apiculture industry. RNA interference (RNAi) technology can be used to inhibit N. ceranae growth by targeting silencing the thioredoxin reductase (TrxR) in N. ceranae. However, suitable carriers are one of the reasons limiting the application of RNAi due to the easy degradation of dsRNA in honeybees. As a vesicle composed of a lipid bilayer, liposomes are a good carrier for nucleic acid delivery, but studies in honeybees are lacking. In this study, liposomes were used for double-stranded RNA (dsRNA) dsTrxR delivery triggering RNAi to inhibit the N. ceranae growth in honeybees. Compared to naked dsTrxR, liposome-dsTrxR reduced N. ceranae numbers in the midgut and partially restored midgut morphology without affecting bee survival and gut microbial composition. The results of this study confirmed that liposomes could effectively protect dsRNA from entering the honeybee gut and provide a reference for using RNAi technology to suppress honeybee pests and diseases.

Mobile agents are agents that can migrate from host-to-host to work in a heterogeneous network environment. A mobile agent can migrate from host-to-host in its plan with the statistics generated on each host through a route known as migration pattern. Migration pattern therefore is the route the agents use to travel within the plan from the first host to the last host. However, there is a need for a comparison between the commonly used optimization algorithms in developing migration patterns for mobile agents with respect to some evaluation metrics. In this paper, the three techniques firefly algorithm (FFA), honeybee optimization (HBO) and particle swarm optimization (PSO) were used for developing migration patterns for mobile agents and their comparison was done based on migration time, time complexity and network load as metrics. PSO is discovered to perform better in terms of network load with an average of 242.3905 bits per second (bps), time complexity with an average of 41.2688 number of nodes (n), and migration/transmission time with an average of 4.203462 seconds (s). [ABSTRACT FROM AUTHOR]

Temperature affects the distribution and success of Apis mellifera subspecies. To survive high temperatures in their natural habitats, unique adaptive traits have been evolved by various A. mellifera subspecies. A. mellifera jemenitica is a sub-tropical subspecies that is native to the Arabian Peninsula. It is one of the most thermotolerant A. mellifera subspecies and the only subspecies that can successfully survive long summers with daily temperatures exceeding 45°C in the Arabian Peninsula. In hot, dry conditions and short seasons, A. m. jemenitica can successfully establish its colonies. In this article, the different morphological, behavioural and physiological aspects of the thermal adaptation of A. m. jemenitica, which are its size, pigmentation, brood cell size, seasonal cycle, foraging behaviour, heat shock proteins and life genes are discussed. These aspects account for how A. m. jemenitica can withstand and survive the high summer temperatures on the Arabian Peninsula.

ABSTRACTSpecies Distribution Models (SDM) play an essential role in establishing factors affecting honeybee species spatial population distribution and ensuring effective niche habitat protection. This study aimed to assess the precision of the triangulation bee lining technique in locating Apis mellifera scutellata colonies within wild habitats, locate A. m. scutellata colony site population distribution and develop a predictive model of A. m. scutellata habitat suitability using SDM algorithms within Hurugwe Safari Area (HSA)-Rifa section, northern Zimbabwe. A survey of A. m. scutellata was carried out between February and October 2019. The triangulation bee lining technique was used to track wild bees to their colony sites. The bee lining technique was evaluated to assess its precision in locating honeybee colonies. Ensemble models were used to develop a predictive suitability niche ecosystem for A. m. scutellata nesting sites. The study located wild honeybee colonies using the triangulation bee lining technique with an 18% accuracy. Bee colonies were found at a mean distance of 27.7 m ± 2.06 from triangulated points. The predicted suitable areas were primarily areas close to the water sources and low-lying areas. The study recommended the use of the triangulation honeybee hunting method in locating wild honeybee colonies for research purposes, wild bee colony inspection for pests and diseases and sustainable organic honey collection.

Byline: M. Keskin, S. Keskin and S. Karlidag Keywords: Apis mellifera L., apitoxin, collagenase, myeloperoxidase, wound healing. INTRODUCTION The honey bee (Apis mellifera L.) is a social insect and it [...]

To identify viruses and compare their abundance levels in the venom glands of hymenopteran species, we conducted venom gland‐specific transcriptome assemblies and analyses of 22 aculeate bees and wasps and identified the RNA genomes of picornaviruses. Additionally, we investigated the expression patterns of viruses in the venom glands over time following capture. Honeybee‐infecting viruses, including the black queen cell virus (BQCV), the deformed wing virus (DWV) and the Israeli acute paralysis virus (IAPV), were highly expressed in the venom glands of Apis mellifera and social wasps. This finding suggests that the venom of bees and wasps is likely to contain these viruses, which can be transmitted horizontally between species through stinger use. Apis mellifera exhibited an increasing pattern of abundance levels for BQCV, DWV, IAPV and Triatovirus, whereas the social wasp Vespa crabro showed increasing abundance levels of IAPV and Triatovirus over different capture periods. This suggests that the venom glands of honeybees and wasps may provide suitable conditions for active viral replication and may be an organ for virus accumulation and transmission. Some viral sequences clearly reflected the phylogeny of aculeate species, implying host‐specific virus evolution. On the other hand, other viruses exhibited unique evolutionary patterns of phylogeny, possibly caused by specific ecological interactions. Our study provides insights into the composition and evolutionary properties of viral genes in the venom glands of certain aculeate bees and wasps, as well as the potential horizontal transmission of these viruses among bee and wasp species. [ABSTRACT FROM AUTHOR]

The study objective was to estimate the genetic structure of the aboriginal honeybee population of Apis mellifera mellifera from the Southern Urals (Russia). We identified in 56 wild and 93 managed colonies two spatially separated haplotypes of the mitochondrial locus COI-COII. Some individuals (17.5%) were represented by the invaded subspecies haplotype. Positive significant autocorrelation and a spatial genetic structure were observed at distance of 0–15 km on alleles of nine microsatellite loci. We did not find a difference in the admixture in the nuclear genes for individuals of alternative haplotypes, which proves a strong introgression process. Considering these results, we recommend reinforced measures to conserve this population. [ABSTRACT FROM AUTHOR]

A total of 151 pollen loads were recovered directly from the honeycombs of Apis dorsata (Rock Bee) collected during December 16, 2011-November 30, 2012, from Nandgaon, Ratnapur, and Lonwahi forest area of Sindewahi Tehsil of Chandrapur District of Maharashtra State, were analyzed. Twelve (7.94%) pollen loads were found to be uniforal, 47 (31.12%) bifloral, and 92 (60.92%) multifloral. The unifloral pollen loads were contained four of Cajanus cajan, three of Capsicum annum, two of Hyptis suaveolens and Prosopis juliflora, and one of Tridax procumbens. The pollen of C. annum was recovered from 96 (63.57%) of the total pollen loads studied. The study highlights C. annum (Solanaceae) do the major pollen sources and C. cajan (Papillionaceae), Lathyrus sativus (Papillionaceae), T. procumbens (Asteraceae), H. suaveolens (Lamiaceae), P. juliflora (Mimosaceae), Blumea spp. (Asteraceae), Capparis grandis (Capparidaceae), and Bidens pilosa (Asteraceae) as fairly important sources of pollen for the honeybees during the summer period. [ABSTRACT FROM AUTHOR]

In the context of global warming, the focus on applying and researching double–skin facade (DSF) systems to reduce energy consumption in buildings has significantly increased. However, researchers have not thoroughly examined the performance and applicability of DSFs in severe cold regions with high winter heating demands. This study aims to evaluate the potential application of DSFs in the harsh cold cities of Northwest China and investigate their role in enhancing energy efficiency in large public buildings. Through energy consumption simulation and a comprehensive evaluation using the TOPSIS entropy weight method, the effects of applying 20 DSF schemes in four cold cities in Xinjiang (Kashgar, Urumqi, Altay, and Turpan) were analyzed. The experimental results indicate that the average EUI energy–saving rates in Kashgar, Urumqi, Altay, and Turpan are 64.75%, 63.19%, 56.70%, and 49.41%, respectively. South–facing orientation is deemed optimal for DSF in Xinjiang cities, with the highest energy–saving rate reaching 15.19%. In Kashgar, the energy–saving benefits of west–facing DSF surpass those of north–facing DSF. Conversely, the order of orientation benefits for other cities is south, north, west, and east. An analysis of heating, cooling, and lighting energy consumption reveals that Box Windows exhibit superior heating energy efficiency, while Corridors are more effective for cooling. This characteristic is also evident in the optimal installation orientation of various types of curtain walls. Given the relatively higher demand for heating compared to cooling in urban areas, Box Windows yields significant benefits when facing south, west, or north; conversely, if there is a high demand for urban cooling, Corridors should be considered in these three directions. Multistorey DSF systems are suitable for east–facing buildings in Xinjiang cities. Selecting suitable DSF schemes based on specific conditions and requirements can reduce building energy consumption. The research findings offer theoretical guidance for designing and implementing DSF in diverse cities in cold regions. [ABSTRACT FROM AUTHOR]

Against a backdrop of declining bee colony health, this study aims to gain a better understanding of the impact of an antimicrobial (Fumidil B®, Can-Vet Animal Health Supplies Ltd., Guelph, ON, Canada) and a probiotic (Bactocell®, Lallemand Inc., Montreal, QC, Canada) on bees' microbiota and the health of their colonies after wintering. Therefore, colonies were orally exposed to these products and their combination before wintering in an environmental room. The results show that the probiotic significantly improved the strength of the colonies in spring by increasing the total number of bees and the number of capped brood cells. This improvement translated into a more resilient structure of the gut microbiota, highlighted by a more connected network of interactions between bacteria. Contrastingly, the antimicrobial treatment led to a breakdown in this network and a significant increase in negative interactions, both being hallmarks of microbiota dysbiosis. Although this treatment did not translate into a measurable colony strength reduction, it may impact the health of individual bees. The combination of these products restored the microbiota close to control, but with mixed results for colony performance. More tests will be needed to validate these results, but the probiotic Bactocell® could be administrated as a food supplement before wintering to improve colony recovery in spring. [ABSTRACT FROM AUTHOR]

Verifying the inclusivity of molecular detection methods gives indications about the reliability of viral infection diagnosis because of the tendency of viral pathogens to undergo sequence variation. This study was aimed at selecting inclusive probes based on reverse transcription–quantitative PCR (RT-qPCR) assays for the diagnosis of the most widespread and detrimental viruses infecting honeybees, namely the acute bee paralysis virus (ABPV), the black queen cell virus (BQCV), the chronic paralysis bee virus (CBPV), the deformed wing virus variants A (DWVA) and B (DWVB), and the sacbrood virus (SBV). Therefore, previously described detection methods were re-evaluated in silico for their specificity and inclusivity. Based on this evaluation, selected methods were modified, or new ones were designed and tested in duplex RT-qPCR reactions. The limits of detection (LODs), effect of multiplexing on sensitivity and the viral RNA quantification potential in bees and hive debris were assessed. This study made available diagnostic assays able to detect an increased number of virus variants compared with previously described tests and two viral pathogens in a single PCR reaction. [ABSTRACT FROM AUTHOR]

Background: The gut microbiota and their hosts profoundly affect each other's physiology and evolution. Identifying host-selected traits is crucial to understanding the processes that govern the evolving interactions between animals and symbiotic microbes. Current experimental approaches mainly focus on the model bacteria, like hypermutating Escherichia coli or the evolutionary changes of wild stains by host transmissions. A method called atmospheric and room temperature plasma (ARTP) may overcome the bottleneck of low spontaneous mutation rates while maintaining mild conditions for the gut bacteria. Results: We established an experimental symbiotic system with gnotobiotic bee models to unravel the molecular mechanisms promoting host colonization. By in vivo serial passage, we tracked the genetic changes of ARTP-treated Snodgrassella strains from Bombus terrestris in the non-native honeybee host. We observed that passaged isolates showing genetic changes in the mutual gliding locus have a competitive advantage in the non-native host. Specifically, alleles in the orphan mglB, the GTPase activating protein, promoted colonization potentially by altering the type IV pili-dependent motility of the cells. Finally, competition assays confirmed that the mutations out-competed the ancestral strain in the non-native honeybee gut but not in the native host. Conclusions: Using the ARTP mutagenesis to generate a mutation library of gut symbionts, we explored the potential genetic mechanisms for improved gut colonization in non-native hosts. Our findings demonstrate the implication of the cell mutual-gliding motility in host association and provide an experimental system for future study on host-microbe interactions. ArN8Xjm3N6Dubzus4DCSQm Video Abstract [ABSTRACT FROM AUTHOR]

Simple Summary: Explanations for the evolution of menopause in humans and other animals concern the indirect fitness benefits that older females can receive by caring for the offspring of their children and grandchildren. Knowledge about the post-reproductive lifespan is still incomplete and until recently was documented only in nonhuman primates, a few species of toothed whales, and guppies, though it has also been documented in insects for gall-forming aphids and parthenogenetic ants. In our paper, we provide evidence that menopause also exists in honeybee societies. Honeybee workers are usually sterile. Nevertheless, the absence of a queen and her pheromones in a nest stimulates workers to activate their ovaries and lay unfertilized eggs that develop into males. However, even in queenless colonies, some workers do not activate their ovaries. In this study, we show that physiologically older workers have less activated ovaries than physiologically younger individuals, indicating that the possibility of workers' reproduction decreases with workers' physiological age. This may happen because bees require a minimum of two weeks to activate their ovaries, meaning physiologically older workers can die before producing their own eggs. Thus, investing energy in helping relatives care for their offspring rather than channelling that energy into one's own reproduction can positively influence workers' fitness. The post-reproductive lifespan is an evolutionary enigma because the cessation of reproduction in animals seems contrary to the maximization of Darwinian fitness. Several theories aim to explain the evolution of menopause, one of which suggests that females of a certain age receive more fitness benefits via indirect selection (kin selection) than they would directly from continuing reproduction. Post-reproductive lifespans are not very common in nature but have been described in humans, nonhuman primates, a few species of toothed whales, guppies, and in some insect societies consisting of clonal colony members, such as aphid and ant societies. Here, we provide evidence that menopause also exists in honeybee societies. Our study shows that workers with a short life expectancy (older and/or injured workers) invest fewer resources and less time in their own reproduction than workers with a long life expectancy (younger and/or uninjured workers), even if their colony is hopelessly queenless. These results are consistent with the kin selection explanation for the evolution of menopause and help us understand the net effects of relatedness and social cooperation in animals. [ABSTRACT FROM AUTHOR]

Simple Summary: Bees are important pollinators that play a role in balancing ecosystems; however, their survival rates have decreased due to many factors, including pathogens and exposure to pesticides. Bees have native mechanisms to help them tackle such challenges, and yet, these may not be enough. To this end, gut symbionts are beneficial, as they can help stimulate bees' immune systems and detoxify ingested toxic chemicals. To enhance the efficiency of these mechanisms, genetic engineering is proposed in this work to further optimize the ability of bee gut symbionts, particularly in the dominant bacteria Snodgrassella alvi and Gilliamella apicola. Engineering strategies are discussed according to the gut symbiotic bacteria's main roles in digestion, essential nutrient provision, and pesticide detoxification. Bees play a crucial role as pollinators, contributing significantly to ecosystems. However, the honeybee population faces challenges such as global warming, pesticide use, and pathogenic microorganisms. Promoting bee growth using several approaches is therefore crucial for maintaining their roles. To this end, the bacterial microbiota is well-known for its native role in supporting bee growth in several respects. Maximizing the capabilities of these microorganisms holds the theoretical potential to promote the growth of bees. Recent advancements have made it feasible to achieve this enhancement through the application of genetic engineering. In this review, we present the roles of gut symbionts in promoting bee growth and collectively summarize the engineering approaches that would be needed for future applications. Particularly, as the engineering of bee gut symbionts has not been advanced, the dominant gut symbiotic bacteria Snodgrassella alvi and Gilliamella apicola are the main focus of the paper, along with other dominant species. Moreover, we propose engineering strategies that will allow for the improvement in bee growth with listed gene targets for modification to further encourage the use of engineered gut symbionts to promote bee growth. [ABSTRACT FROM AUTHOR]

Temperature aff ects the distribution and success of Apis mellifera subspecies. To survive high temperatures in their natural habitats, unique adaptive traits have been evolved by various A. mellifera subspecies. A. mellifera jemenitica is a subtropical subspecies that is native to the Arabian Peninsula. It is one of the most thermotolerant A. mellifera subspecies and the only subspecies that can successfully survive long summers with daily temperatures exceeding 45°C in the Arabian Peninsula. In hot, dry conditions and short seasons, A. m. jemenitica can successfully establish its colonies. In this article, the diff erent morphological, behavioural and physiological aspects of the thermal adaptation of A. m. jemenitica, which are its size, pigmentation, brood cell size, seasonal cycle, foraging behaviour, heat shock proteins and life genes are discussed. These aspects account for how A. m. jemenitica can withstand and survive the high summer temperatures on the Arabian Peninsula. [ABSTRACT FROM AUTHOR]

Micronutrient deficiency is emerging as a significant public health concern in India. Facilitating pollinator populations in agroecosystems could be an efficient mechanism to ensure food production, both quantitatively and qualitatively. To better understand the role of pollinators in the agroecosystems of India, we conducted an analysis of crop production based on their reliance on pollinators during the period 2010–2021. Our findings indicate that although there were no significant changes in the yearly production or cultivation area of various pollinator‐dependent crops, the average yearly rate of crop‐yield increase in pollinator‐dependent crops was notably lower than in crops that did not depend on pollinators during 2015–2021. The study highlights that a significant portion of vitamins, particularly B7, B9, C and K, and carotenoids come from pollinator‐dependent crops. The findings of this study, which highlight the considerable contribution of micronutrients originating from pollinator‐dependent crops, are consistent with results observed in the Republic of Korea. The loss of pollinator populations may result in an approximately 19% deficit in vitamin C. Although vegetables and fruits contain substantial quantities of minerals per unit weight, a significant portion of these minerals is sourced from non‐pollinator‐dependent crops, particularly cereals and pulses, owing to their abundance. We propose that pollinator‐friendly habitat management could be a sustainable solution to avoid the negative consequences associated with reduced food and nutrition arising from a lack of pollinators in agroecosystems. [ABSTRACT FROM AUTHOR]

Despite the country's potential for apicultural resources, native races, productive and reproductive performances are at relatively low levels. Numerous factors, including environmental factors and honeybee races, may be to blame for this. As a result, this study was carried out to assess local honeybee race performance and choose the best colonies in the various agro ecologies of the study area. For this purpose, 90 colonies of Apis mellifera monticolla (30 colonies per agro ecology under on farm) were kept in improved box hives and assessed for egg area, larvae area, pupae area, nectar and honey area, as well as honey yield and swarming tendency. Egg sizes in the research area range from 519.25cm2 to 1343.80cm2, while larvae and pupae have respective sizes of 574.07cm2 to 1407.05cm2 and 618.76 to 1471.83cm2. Dessie zuriya district had the highest pollen area, nectar area, honey area, and honey yield (15.28cm2, 343.17cm2, 386.13cm2, and 22.13cm2), followed by Kalu (8.97cm2, 236.02cm2, and 18.88cm2), and Tehuledere (6.01cm2, 142.54cm2, 150.39cm2, and 12.83cm2, correspondingly). The development of queen cells and swarming behavior were unaffected by agro ecology. In general, compared to other ecotypes and races in the country, honeybee colonies in the study area generally performed better in terms of both productive and reproductive aspects. In this study, colonies located in the zone's highland performed better than colonies located in the study area's midland and lowland. [ABSTRACT FROM AUTHOR]

Members of the Geminviridae family are circular single-stranded DNA plant-infecting viruses, some of which impact global food production. Geminiviruses are vectored by sap-feeding insects such as leafhoppers, treehoppers, aphids, and whiteflies. Additionally, geminivirus sequences have also been identified in other insects such as dragonflies, mosquitoes, and stingless bees. As part of a viral metagenomics study on honeybees and solitary bees (Nomia sp.), two geminivirus genomes were identified. These represent a novel citlodavirus (from honeybees collected from Westmoreland, Jamaica) and a mastrevirus-like genome (from a solitary bee collected from Tempe, Arizona, USA). The novel honeybee-derived citlodavirus genome shares ~61 to 69% genome-wide nucleotide pairwise identity with other citlodavirus genome sequences and is most closely related to the passion fruit chlorotic mottle virus identified in Brazil. Whereas the novel solitary bee-derived mastrevirus-like genome shares ~55 to 61% genome-wide nucleotide identity with other mastreviruses and is most closely related to tobacco yellow dwarf virus identified in Australia, based on pairwise identity scores of the full genome, replication-associated protein, and capsid protein sequences. Previously, two geminiviruses in the Begomovirus genus were identified in samples of stingless bee (Trigona spp.) samples. Here, we identify viruses that represent two new species of geminiviruses from a honeybee and a solitary bee, which continues to demonstrate that plant pollinators can be utilized for the identification of plant-infecting DNA viruses in ecosystems. [ABSTRACT FROM AUTHOR]

Bee venom contains several bioactive components, including enzymatic and non-enzymatic proteins. There is increasing interest in the bioactive components of bee venom since they have exhibited various pharmacological effects. Recently, Apis mellifera waprin (Amwaprin) was identified as a novel protein in Apis mellifera (honeybee) venom and characterized as an antimicrobial agent. Herein, the novel biological function of Amwaprin as an antioxidant is described. In addition, the antioxidant effects of Amwaprin in mammalian cells were investigated. Amwaprin inhibited the growth of, oxidative stress-induced cytotoxicity, and inflammatory response in mammalian NIH-3T3 cells. Amwaprin decreased caspase-3 activity during oxidative stress and exhibited protective activity against oxidative stress-induced cell apoptosis in NIH-3T3 and insect Sf9 cells. The mechanism underlying the cell protective effect of Amwaprin against oxidative stress is due to its direct binding to the cell membrane. Furthermore, Amwaprin demonstrated radical-scavenging activity and protected against oxidative DNA damage. These results suggest that the antioxidant capacity of Amwaprin is attributed to the synergistic effects of its radical-scavenging action and cell shielding, indicating its novel role as an antioxidant agent. [ABSTRACT FROM AUTHOR]

Positive and negative experiences can alter animal brain dopamine levels.1 When first arriving at a rewarding food source or beginning to waggle dance and recruit nestmates to food, honeybees have increased brain dopamine levels, indicating a desire for food.2 We provide the first evidence that an inhibitory signal, the stop signal, which counters waggle dancing and is triggered by negative events at the food source, can decrease head dopamine levels and dancing, independent of the dancer having any negative experiences. The hedonic value of food can therefore be depressed simply by the receipt of an inhibitory signal. Increasing the brain dopamine levels reduced the aversive effects of an attack, increasing the time that bees spent subsequently feeding and waggle dancing and decreasing their stop signaling and time spent in the hive. Because honeybees regulate food recruitment and its inhibition at the colony level, these results highlight the complex integration of colony information with a basic and highly conserved neural mechanism in mammals and insects.2 VIDEO ABSTRACT.

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.

Social insects, such as ants, termites, and honeybees, have evolved sophisticated societies where collaboration and division of labor enhance survival of the whole colony, and are thus considered “superorganisms”. Historically, studying behaviors involving large groups under natural conditions posed significant challenges, often leading to experiments with a limited number of organisms under artificial laboratory conditions that incompletely reflected the animals’ natural habitat. A promising approach to exploring animal behaviors, beyond observation, is using robotics that produce stimuli to interact with the animals. However, their application has predominantly been constrained to small groups in laboratory conditions. Here we present the design choices and development of a biocompatible robotic system intended to integrate with complete honeybee colonies in the field, enabling exploration of their collective thermoregulatory behaviors via arrays of thermal sensors and actuators. We tested the system’s ability to capture the spatiotemporal signatures of two key collective behaviors. A 121-day observation revealed thermoregulation activity of the broodnest area during the foraging season, followed by clustering behavior during winter. Then we demonstrated the system’s ability to influence the colony by guiding a cluster of bees along an unnatural trajectory, via localized thermal stimuli emitted by two robotic frames. These results showcase a system with the capability to experimentally modulate honeybee colonies from within, as well as to unobtrusively observe their dynamics over extended periods. Such biohybrid systems uniting complete societies of thousands of animals and interactive robots can be used to confirm or challenge the existing understanding of complex animal collectives.

Interactions between environmental stressors may contribute to ongoing pollinator declines, but have not been extensively studied. Here, we examined the interaction between the agricultural fungicide Pristine (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin) and high temperatures on critical honeybee behaviours. We have previously shown that consumption of field-realistic levels of this fungicide shortens worker lifespan in the field and impairs associative learning performance in a laboratory-based assay. We hypothesized that Pristine would also impair homing and foraging behaviours in the field, and that an interaction with hot weather would exacerbate this effect. Both field-relevant Pristine exposure and higher air temperatures reduced the probability of successful return on their own. Together, the two factors synergistically reduced the probability of return and increased the time required for bees to return to the hive. Pristine did not affect the masses of pollen or volumes of nectar or water brought back to the hive by foragers, and it did not affect the ratio of forager types in a colony. However, Pristine-fed bees brought more concentrated nectar back to the hive. As both agrochemical usage and heat waves increase, additive and synergistic negative effects may pose major threats to pollinators and sustainable agriculture. [ABSTRACT FROM AUTHOR]