Image background assessment as a novel technique for insect microhabitat identification

Native insect flower visitors can be important contributors to crop pollination, yet little is known of their pollination abilities and the resources (habitat) they need to be supported within crop agroecosystems. Here, we compared the abundance and pollination abilities of the golden drone fly (Eristalinus punctulatus) to the European honey bee (Apis mellifera) in hybrid carrot crop fields known to produce variable seed yields in regional New South Wales, Australia. We further observed the egg‐laying behaviours of female golden drone flies at a commercial berry orchard to provide insight into the habitat needs of this species. In hybrid carrot crop fields, golden drone flies were far less abundant flower visitors than European honey bees; however, these flies deposited more carrot pollen grains on average (8.21 ± 3.04 SE) onto carrot flowers than European honey bees (3.45 ± 1.06 SE). Both insects also deposited pollen onto a similar number of carrot flowers (pollinated) per visit (about 2 out of 18). Golden drone flies were observed laying eggs within masses of discarded red raspberry plant roots and soil (root balls) at a commercial berry orchard. The natural habitat utilised by these flies, as well as their egg‐laying behaviours, were described for the first time. Our results indicate that golden drone flies are effective pollinators of hybrid carrot crop plants. The habitat that these flies utilised to lay eggs (discarded plants and water) is cheap and commonly found in crop agroecosystems. Therefore, we recommend placing this low‐cost habitat within, or nearby, crop fields as a potential management practice to support the lifecycle needs of golden drone flies and other non‐bee pollinators.

Worker honeybees (Apis mellifera) usually only lay eggs when their colony is queenless. However, an extremely rare 'anarchistic' phenotype occurs, in which workers develop functional ovaries and lay large numbers of haploid eggs which develop into adult drones despite the presence of the queen. Studies of such colonies can give important insights into the mechanisms by which worker sterility is maintained in normal colonies. Here we report on the results of a breeding programme which enhanced the frequency of the anarchistic phenotype. Colonies derived from queens inseminated only by worker-laid males showed up to 9% of workers with highly developed ovaries. In these colonies a large proportion of males arose from worker-laid eggs. Colonies headed by queens inseminated with 50% worker-laid drones and 50% queen-laid drones showed variable phenotypes. In most such colonies there was no worker reproduction. In some, many workers had highly developed ovaries, but no worker-laid eggs were reared. In one colony, many worker-laid eggs were reared to maturity. The results suggest that the anarchy phenotype results from a complex interaction of queen genotype, the worker genotype of subfamilies that successfully reproduce and of those that do not, and the external environment.

IntroductionIn Australia, as well as many other regions of the world, European honey bees Apis mellifera are an introduced species and may harm native bee fauna by competing with them for food resources. Field studies have revealed negative associations between honey bee and native bee abundance, but whether this translates to fitness costs for native bees is unclear.MethodsUsing drilled wooden-block trap nests, we evaluated whether honey bee abundance is associated with fitness parameters (number of nests, provisioned cells per nest, offspring number, mortality rate, sex ratio, and body size) of cavity-nesting native bees over 2 years. We also conducted palynological analyses to measure pollen resource overlap and evaluate whether this impacts native bee fitness.Results and discussionGreater honey bee abundance was associated with a male-biased sex ratio in the native bee progeny across years and an increased mortality rate of native bee progeny in the first year. Most non-significant associations were also in the directions predicted from honey bees adversely impacting native bee fitness. In the first year, greater pollen morphospecies overlap was associated with fewer provisioned cells. In conclusion, we demonstrated that honey bees have the potential to have harmful consequences for native bee fitness.

Many crop plants rely on insect pollination, particularly insect-pollinated crops which are functionally dioecious. These crops require insects to move pollen between separate plants which are functionally male or female. While honey bees are typically considered the most important crop pollinator species, many other insects are known to visit crops but the pollination contribution of the full diversity of these flower visitors is poorly understood. In this study, we examine the role of diverse insect pollinators for two kiwifruit cultivars as model systems for dioecious crops: Actinidia chinensis var. deliciosa ‘Hayward’ (a green-fleshed variety) and A. chinensis var. chinensis ‘Zesy002’ (a gold-fleshed variety). In our round-the-clock insect surveys, we identified that psychodid flies and mosquitoes were the second and third most frequent floral visitors after honey bees (Apis mellifera L), but further work is required to investigate their pollination efficiency. Measures of single-visit pollen deposition identified that several insects, including the bees Leioproctus spp. and Bombus spp. and the flies Helophilus hochstetteri and Eristalis tenax, deposited a similar amount of pollen on flowers as honey bees (Apis mellifera). Due to their long foraging period and high pollen deposition, we recommend the development of strategies to boost populations of Bombus spp., Eristalis tenax and other hover flies, and unmanaged bees for use as synergistic pollinators alongside honey bees.

Typing of Melissococcus plutonius isolated from European and Japanese honeybees suggests spread of sequence types across borders and between different Apis species

<p><b>Emerging infectious diseases threaten public health, livestock economies, and wildlife. Human-mediated species introductions can alter host and pathogen communities that shape the dynamics of infectious diseases. Several RNA viruses that have been linked to population declines in wild pollinators and losses of managed honey bees have been detected in multiple other species and are suspected to circulate within insect communities. Yet, we lack an understanding of how disease dynamics are affected by the introduction of novel species. These introduced species include invasive ants, which can disturb honey bees and become a pest in apiaries. The Argentine ant (Linepithema humile) is a globally successful invader that has been observed to attack bees and multiple bee-associated viruses have been detected in this ant species.</b></p> <p>Here, I studied interactions between Argentine ants and European honey bees (Apis mellifera) and how these interactions affect viral dynamics in beehives. I first tested a range of pollinators and associated insects for RNA viruses that are pathogenic to honey bees. Bee-associated viruses showed evidence for active viral replication in several pollinator species but also in species that cohabit in beehives such as ants, spiders, and cockroaches. Using phylogenetic analyses, I found that viral transmission within communities was shaped by geographic origin rather than being restricted by species barriers. Next, I used a longitudinal field study to test whether Argentine ant presence affected pathogen infections and survival in beehives. Argentine ants tested positive for three bee-associated viruses even before beehives were moved into ant-infested sites. Increased levels of deformed wing virus in beehives in autumn were associated with ant presence, although hive mortality was not affected by ants over the duration of this experiment. I used RNA sequencing on a subset of honey bee samples collected during autumn to study the RNA virome and identify transcriptomic responses associated with ant presence. Twelve RNA viruses were found in beehives, among those, three plant-associated viruses and an unclassified RNA virus that had not previously been observed in honey bees. Deformed wing virus showed the highest viral titres in most hives, but was only marginally affected by ant presence. Sacbrood virus and tomato ringspot virus levels were increased in hives with ants, however, both viruses are not known to infect Argentine ants and the plant-associated tomato ringspot virus seems unlikely to affect bee health.</p> <p>Lastly, I tested the feasibility of controlling Argentine ants in apiaries using a novel pest control strategy. RNA interference is a conserved cellular gene regulation mechanism that could be used to silence specific genes in ants. Using double-stranded RNA (dsRNA) to silence two immune-related genes in Argentine ants was expected to increase pathogen susceptibility, which could then lead to higher pathogen levels that reduce ant numbers. My results indicated that no consistent immune silencing could be achieved in the field. Immune gene expression changes were observed, but pathogen titres were not affected, and ant numbers stayed high. Argentine ant control using a conventional insecticide significantly increased bee survival, whereas many hives in the dsRNA and control group abandoned their hives due to ant attacks. Although population control was not successful using the two Argentine ant-specific dsRNAs, insights into ant immunity and ant-bee interactions could improve the development of novel control strategies.</p> <p>Bee-associated viruses have repeatedly been detected in ant species, yet, this is one of the first studies to investigate whether ants affect viral dynamics in honey bees. I showed that invasive Argentine ants are associated with increases in viral pathogens in honey bees. The mechanisms by which ants affect bee disease are unknown, although there is some evidence for ants transmitting viruses or causing stress responses in bees that affect immunity. The findings of this thesis highlight the risk of invasive ant species disrupting pollination services. New and environmentally-friendly methods to control invasive species are urgently needed to improve bee health and limit the spread of invasive ants, such as Argentine ants. The high prevalence of bee-associated viruses and viral diversity in ants suggests that pathogens that are suitable for population control might be present in ant populations, although risks of spillovers into other species need to be carefully considered.</p>

Honey is one of the historical natural products produced by honey bees. Humans used it as medicine and protection against some infectious diseases, a honey quale is dependent in its classification of the honey bee. The most famous accomplished honey bees in the world are Africanized honey bee (Apis mellıfera .l), Western honey bee or European honey bee (Apis mellifera), Eastern honey bee (Apis cerana), Philippine honey bee (Apis nigrocincta), Koschevnikovs honey bee (A. koschevnikovi), Giant Honeybees (Apis dorsata), Dwarf Honeybees (Apis andreniformis). This study collected the health effect, quality and usage of honey in several industries; also argue about the nutritious value of honey that the most important parts are protein, carbohydrates, vitamin, and minerals. Honey is not only a nutrient it also plays a major role in many other products, uses of honey in cosmetics as a protective and softener of skin, it is used up to 22% in paste masks and mud packs (which are considered rinse-off formulations). Uses of honey in Ayurveda for prevention of irritation, cough, healthy teeth, gums, and boons. Production of honey has been compared in 2013 to 2018, in 2013 the world production of honey was 1,664 thousand tones with an increase of 1/3 in just two decades. China, where manufacturing amounted to 466.3 thousand tons, i.e. approximately 29% of the global volume of output, these are showing that China was the biggest producer of honey in the world but it gets a decrease in 2018 as well as Brazil, America, Russia, Ethiopia, and Iran. And it increases in Turkey, Argentine, India, Mexico, and Poland.

SummaryA one-year study was conducted on nectar foraging and honey storage by Africanized (tropical) and European (temperate) honeybee (Apis mellifera L.) colonies. Patterns of nectar foraging were distinctly different in the two geographical types. Among Africanized honeybees, lower proportions of the foraging force were engaged in foraging for nectar only, but a higher proportion foraged for both nectar and pollen. In addition, Africanized bees collected lower volumes per foraging trip than European honeybees, but returned to the colony with nectars having a higher sugar content. The two geographical types contributed similar amounts of joules (energy) per average forager and, therefore, made similar net contributions of carbohydrate energy to the colony. However, European colonies always had more stored honey than Africanized colonies. The observed differences in foraging patterns could not account for all of the measured difference in stored honey; several testable hypotheses are presented to explain ...

Simple SummaryWing venation traits are used to identify honey bee subspecies. While several wing-based tools are available, they suffer from weaknesses that were addressed by the recently developed software DeepWings©. This software allows fully automated identification of wing images in a friendly, free, and rapid manner. Here, we sought to test DeepWings© on 14,816 wing images representing 2601 colonies sampled in the native areas of three widespread subspecies in Europe: the Iberian honey bee (Apis mellifera iberiensis), the dark honey bee (Apis mellifera mellifera), both belonging to the M lineage, and the Carniolan honey bee (Apis mellifera carnica), belonging to the C lineage. DeepWings© classification of these colonies largely matched the endemic M and C lineages, with proportions of 71.4% and 97.6%, respectively. At the subspecies-level the matching proportions were 89.7% for the Iberian honey bee, 41.1% for the dark honey bee and 88.3% for the Carniolan honey bee, which can be explained by DeepWings© sometimes confounding closely related subspecies and, more importantly, by genetic pollution. A comparison between DeepWings© data and molecular data revealed that the agreement between the two is weaker when there is genetic pollution. Our results suggest that DeepWings© is a valuable tool for honey bee identification, which can be used not only for breeding and conservation but also for research purposes.DeepWings© is a software that uses machine learning to automatically classify honey bee subspecies by wing geometric morphometrics. Here, we tested the five subspecies classifier (A. m. carnica, Apis mellifera caucasia, A. m. iberiensis, Apis mellifera ligustica, and A. m. mellifera) of DeepWings© on 14,816 wing images with variable quality and acquired by different beekeepers and researchers. These images represented 2601 colonies from the native ranges of the M-lineage A. m. iberiensis and A. m. mellifera, and the C-lineage A. m. carnica. In the A. m. iberiensis range, 92.6% of the colonies matched this subspecies, with a high median probability (0.919). In the Azores, where the Iberian subspecies was historically introduced, a lower proportion (85.7%) and probability (0.842) were observed. In the A. m mellifera range, only 41.1 % of the colonies matched this subspecies, which is compatible with a history of C-derived introgression. Yet, these colonies were classified with the highest probability (0.994) of the three subspecies. In the A. m. carnica range, 88.3% of the colonies matched this subspecies, with a probability of 0.984. The association between wing and molecular markers, assessed for 1214 colonies from the M-lineage range, was highly significant but not strong (r = 0.31, p < 0.0001). The agreement between the markers was influenced by C-derived introgression, with the best results obtained for colonies with high genetic integrity. This study indicates the good performance of DeepWings© on a realistic wing image dataset.

European honey bee (Apis mellifera L.,) was introduced in Kashmir during late 19th century and since, then its domestication is continued in diverse conditions of Kashmir region. It appears well adapted to unique climatic conditions of the valley. In order to analyse diversification in its characters during ecological adaptations to different altitudes and climatic conditions, morphological characterization of Apis mellifera was employed at different altitudes of Kashmir region. Samples of honey bees were obtained from nine locations representing three different altitudes viz., high, medium and low of Kashmir. Total of 17 characters were measured at three different altitudes. Highly significant difference (p≤0.01) in 15 characters were observed was observed among three altitudes. The three main characters viz., proboscis length, fore wing length and cubital index were compared with earlier data as reported by Ruttner (1988). The proboscis length (6.35±0.06 mm) at high altitude, forewing length (9.21±0.01 mm) at mid altitude, cubital index at low altitude (2.55±0.08 mm) were found same or close to A. mellifera ligustica. However, other parameters did not support presence of this sub-species, therefore, it can be conclude that A. mellifera might have undergone degradations in its certain characters after its introduction in Kashmir region due to diverse climate conditions.

The gut of the European honey bee (Apis mellifera) possesses a relatively simple bacterial community, but little is known about its community of prophages (temperate bacteriophages integrated into the bacterial genome).Although prophages may eventually begin replicating and kill their bacterial hosts, they can also sometimes be beneficial for their hosts by conferring protection from other phage infections or encoding genes in metabolic pathways and for toxins.In this study, we explored prophages in 17 species of core bacteria in the honey bee gut and two honey bee pathogens.Out of the 181 genomes examined, 431 putative prophage regions were predicted.Among core gut bacteria, the number of prophages per genome ranged from 0 -7 and prophage composition (the compositional percentage of each bacterial genome attributable to prophages) ranged from 0 -7%.Snodgrassella alvi and Gilliamella apicola had the highest median prophages per genome (3.0 ± 1.46; 3.0 ± 1.59), as well as the highest prophage composition (2.58% ± 1.4; 3.0% ± 1.59).The pathogen Paenibacillus larvae had a higher median number of prophages (8.0 ± 5.33) and prophage composition (6.40% ± 3.08) than the pathogen Melissococcus plutonius or any of the core bacteria.Prophage populations were highly specific to their bacterial host species, suggesting most prophages were acquired recently relative to the divergence of these bacterial groups.Furthermore, functional annotation of the predicted genes encoded within the prophage regions indicates that some prophages in the honey bee gut encode additional benefits to their bacterial hosts, such as genes in carbohydrate metabolism.Collectively, this survey suggests that prophages within the honey bee gut may contribute to the maintenance and stability of the honey bee gut microbiome and potentially modulate specific members of the bacterial community, particularly S. alvi and G. apicola.

From bioaccumulation to biodecumulation: Nickel movement from Odontarrhena lesbiaca (Brassicaceae) individuals into consumers

To conserve local biodiversity and ensure the provision of pollination services, it is essential to understand the impact of pesticides on wild honey bees. Most studies that have investigated the effects of pesticides on honey bees have focused on the European honey bee (Apis mellifera (Hymenoptera: Apidae)), which is commonly domesticated worldwide. However, the Asian honey bee (Apis cerana) is widely distributed throughout Asia, and toxicity data are lacking for this species. This study aimed to fill this important knowledge gap. In this study, we determined the acute contact toxicity in A. cerana to various pesticides, including neonicotinoids, fipronil, organophosphorus, synthetic pyrethroids, carbamate, and anthranilic diamide. Based on the test duration of 48 h of contact LD50 tests, A. cerana was most sensitive to dinotefuran (0.0014 μg/bee), followed by thiamethoxam (0.0024 μg/bee) and fipronil (0.0025 μg/bee). Dinotefuran is used extensively in Asia, thereby potentially creating a substantial hazard. More generally, A. cerana was approximately one order of magnitude more sensitive than was A. mellifera to most of the pesticides evaluated. The results of our study suggest that neonicotinoid pesticides should not be considered as a single group that acts uniformly on all honey bees, and that more careful management strategies are required to conserve A. cerana populations than A. mellifera.

Contact toxicities were established for acetone formulations of azinphosmethyl, carbaryl, methyl parathion, and permethrin applied to workers of Africanized and European honey bee (Apis mellifera L.) types. For each insecticide, 95% fiducial limits at the LC50 levels for the two bee types did not overlap. Africanized bees showed greater tolerance to all the chemicals except carbaryl; differences in tolerance to each of the four chemicals were all about 2-fold. The order of toxicity of the compounds on the Africanized bees was permethrin > carbaryl > azinphosmethyl > methyl parathion; on the European honey bees, the order of toxicity was permethrin > azinphosmethyl > carbaryl> methyl parathion. Significant differences between the bee types were noted in the slopes of the probit regressions estimated for three of the compounds. The Africanized bees responded more homogeneously than the European bees to azinphosmethyl; European bees reacted more homogeneously than Africanized bees to carbaryl and permethrin.

Honey bees gather pollen from flowering plants, using it as a vital protein source and, in turn, acquire pollen-associated microbes that interact with their existing gut microbiota. Despite their ecological importance, limited information exists regarding the gut microbiota of African savannah honey bees (Apis mellifera scutellata Lepeletier) and how diet and its associated microbial community influence this crucial internal ecosystem. This study aimed to investigate the differences in gut microbiota between wild honey bees collected during the flowering season and microbially depleted honey bees reared under semi-sterile conditions and fed various protein diets. To achieve this, freshly hatched worker bees were maintained in hoarding cages and assigned one of four protein diets: fresh sunflower pollen, casein, sterilised casein, or sterilised pollen. High-throughput DNA metabarcoding was then employed to compare the microbial composition of the honey bee gut across these groups. Our findings revealed that the gut of microbially depleted honey bees exhibited higher species diversity and richness. Conversely, the non-core gut microbial community predominated in wild bees and those fed the different protein diets. Specifically, Commensalibacter, Bartonella, and Bifidobacterium were the most dominant bacterial genera across all treatments. Interestingly, Gilliamella, a common core gut bacterium, was undetected, while Apibacter was exclusively found in wild honey bees. Furthermore, pollen-associated microbes such as Devosia and Pedobacter were identified solely in the gut of honey bees fed a pollen diet. Functional predictions of the gut microbial community also indicated the presence of key enzymes such as β-glucosidase, β-galactosidase, pyruvate dehydrogenase and phosphoglycerate mutase, which are crucial for enhancing nutrient absorption, digestion, and carbohydrate metabolism. These results underscore the intricate relationship between honey bees, microbes, and plants, offering valuable insights into how diet and its associated microbial communities could shape the gut microbiota of African honey bees. KEY POINTS: • The non-core gut microbiota dominates the African savannah honey bee •The type of diet influenced the microbial diversity and community abundance in the honey bee gut •Key enzymes involved in digestion, nutrition absorption, and carbohydrate metabolism were enhanced in the gut •Pollen-associated microbes found in the diet present potential avenues for probiotic development to improve honey bee health.