Apis Cerana Research Articles

Whole-Genome Resequencing-Based Qualitative Trait Locus Mapping Correlated yellow with the Mutant Color in Honeybees, Apis cerana cerana.

The honeybee, Apis cerana cerana (Ac), is an important pollinator and has adapted to the local ecological environment with relevant coloration. The cuticle coloration of the brown (br) mutant is brown instead of black in wild-type individuals. Therefore, this study aimed to identify and characterize the gene responsible for the br mutation. Genome resequencing with allele segregation measurement using Euclidean distance followed by Lowess regression analysis revealed that the color locus linked to the mutation was located on chromosome 11. A 2-base deletion on exon 4 was identified in the g7628 (yellow) gene after genome assembly and sequence cloning. In addition, the cuticle color of the abdomen of worker bees changed from black to brown when a defect was induced in the yellow gene using short interfering RNA (siRNA); however, the survival rate did not decrease significantly. These results indicate that the yellow gene participated in the body pigmentation, and its defect was responsible for the br mutation. This study promotes the understanding of the molecular basis of body coloration in honeybees, enriching the molecular mechanisms underlying insect pigmentation.

Japanese honey bees (Apis cerana japonica) have swarmed more often over the last two decades

The impacts of temperature increase are a concern for honey bees, which are major pollinators of crops and wild plants. Swarming is the reproductive behavior of honey bees that increases colony numbers. Honey bee colonies sometimes swarm multiple times, with each swarming termed a “swarming event” and a series of these events called a “swarming cycle.” The number of swarming events per swarming cycle varies widely depending on climatic conditions and subspecies, and the recent temperature increase due to global warming might be affecting the number of swarming events per swarming cycle of native honey bees. We clarified long-term changes in the number of swarming events per swarming cycle of Japanese honey bees (Apis cerana japonica) by collecting beekeepers’ swarming logbooks. The survey showed that between 2000 and 2022, Japanese honey bees swarmed 1 to 8 times per swarming cycle. Generalized linear model analysis indicated that year had a significant positive effect (coefficient, 0.03; 95% CI, 0.01–0.04); that is, the number of swarming events per swarming cycle showed a moderate increase over time. In addition, we found that colonies swarmed more often in a cycle when the swarming process began in early spring, especially in March. Considering the notably strong trend in Japan of warmer temperatures in March, the number of swarming events per swarming cycle may be increasing because reproduction is beginning earlier in the year. Further analyses are needed to verify the causal relationship of temperature increase on the number of swarming events per swarming cycle.

1 H-NMR revealed pyruvate as a differentially abundant metabolite in the venom glands of Apis cerana and Apis mellifera.

As a common defense mechanism in Hymenoptera, bee venom has complex components. Systematic and comprehensive analysis of bee venom components can aid in early evaluation, accurate diagnosis, and protection of organ function in humans in cases of bee stings. To determine the differences in bee venom composition and metabolic pathways between Apis cerana and Apis mellifera, proton nuclear magnetic resonance (1 H-NMR) technology was used to detect the metabolites in venom samples. A total of 74 metabolites were identified and structurally analyzed in the venom of A. cerana and A. mellifera. Differences in the composition and abundance of major components of bee venom from A. cerana and A. mellifera were mapped to four main metabolic pathways: valine, leucine and isoleucine biosynthesis; glycine, serine and threonine metabolism; alanine, aspartate and glutamate metabolism; and the tricarboxylic acid cycle. These findings indicated that the synthesis and metabolic activities of proteins or polypeptides in bee venom glands were different between A. cerana and A. mellifera. Pyruvate was highly activated in 3 selected metabolic pathways in A. mellifera, being much more dominant in A. mellifera venom than in A. cerana venom. These findings indicated that pyruvate in bee venom glands is involved in various life activities, such as biosynthesis and energy metabolism, by acting as a precursor substance or intermediate product.

Post-invasion selection acts on standing genetic variation despite a severe founding bottleneck

Invasive populations often have lower genetic diversity relative to the native-range populations from which they derive.1,2 Despite this, many biological invaders succeed in their new environments, in part due to rapid adaptation.3,4,5,6 Therefore, the role of genetic bottlenecks in constraining the adaptation of invaders is debated.7,8,9,10 Here, we use whole-genome resequencing of samples from a 10-year time-series dataset, representing the natural invasion of the Asian honey bee (Apis cerana) in Australia, to investigate natural selection occurring in the aftermath of a founding event. We find that Australia's A.cerana population was founded by as few as one colony, whose arrival was followed by a period of rapid population expansion associated with an increase of rare variants.11 The bottleneck resulted in a steep loss of overall genetic diversity, yet we nevertheless detected loci with signatures of positive selection during the first years post-invasion. When we investigated the origin of alleles under selection, we found that selection acted primarily on the variation introduced by founders and not on the variants that arose post-invasion by mutation. In all, our data highlight that selection on standing genetic variation can occur in the early years post-invasion, even where founding bottlenecks are severe.

The impact of climate change and the conservation of the keystone Asian honeybee using niche models and systematic prioritization.

Global warming has seriously disturbed the Earth's ecosystems, and in this context, Asian honeybee (Apis cerana) has experienced a dramatic decline in recent decades. Here, we examined both direct and indirect effects of climate change on A. cerana through ecological niche modeling of A. cerana, and its disease pathogens (i.e., Chinese sacbrood virus and Melissococcus plutonius) and enemies (i.e., Galleria mellonella and Vespa mandarinia). Ecological niche modeling predicts that climate change will increase the potential suitability of A. cerana, but it will also cause some of the original habitat areas to become unsuitable. Outbreak risks of Chinese sacbrood disease and European Foulbrood will increase dramatically, while those of G. mellonella and V. mandarinia will decrease only slightly. Thus, climate change will produce an unfavorable situation for even maintaining some A. cerana populations in China in the future. Genetic structure analyses showed that the A. cerana population from Hainan Island had significant genetic differentiation from that of the mainland, and there was almost no gene flow between the 2, suggesting that urgent measures are needed to protect the unique genetic resources there. Through taking an integrated planning technique with the Marxan approach, we optimized conservation planning, and identified potential nature reserves (mainly in western Sichuan and southern Tibet) for conservation of A. cerana populations. Our results can provide insights into the potential impact of climate change on A. cerana, and will help to promote the conservation of the keystone honeybee in China and the long-term sustainability of its ecosystem services.

Diversity and foraging activity of coffee insect pollinators in land near and far from the forest of North Sumatra, Indonesia

Abstract. Sinaga RR, Maryana N, Hidayat P. 2024. Diversity and foraging activity of coffee insect pollinators in land near and far from the forest of North Sumatra, Indonesia. Biodiversitas 25: 240-248. Coffee (Coffea spp.) is among the plant species that depend on insects to aid in its pollination process, where insect pollinators serve as intermediaries facilitating plant pollination. This study examined the diversity, abundance, and visiting activity of insect pollinators in coffee cultivation located in different areas, specifically those near the forest and those far from the forest in Simalungun District, North Sumatra. The study was carried out from September 2022 to February 2023 in six coffee cultivation sites spread across four sub-districts in Simalungun District, i.e., Silimakuta, Pematang Silimakuta, Purba, and Dolok Silau. The study's results revealed the presence of 16 species of insect pollinators in coffee cultivation near the forest and 13 species in those located far from the forest. The identified insect pollinators species were from Order Hymenoptera; Apis cerana, Lasioglossum sp., Dolichovespula norvegicoides, Diptera; Stomorhina discolor, Musca domestica, Bactrocera papayae, Nephrotoma flavescens, Episyrphus viridaureus, Oebalia sp., Coleoptera; Pachnaeus sp., Cantharis sp., Scarabaeidae sp 1., Maladera japonica, Lepidoptera; Agrotis segetum, Bradina diagonalis, and Nyctemera baulus. Notably, the abundance of coffee insect pollinators was more pronounced in close proximity to the forest compared to areas far from the forest. Regarding the visiting duration of these insects, S. discolor exhibited the longest visit duration of 76.46±1.19 seconds per flower, and the shortest duration in A. cerana was recorded at 8.35±0.23 seconds per flower.

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.

Impact of Brood Cell Cocoons on Metal Accumulation and CYP450 Detoxification Gene Expression in Apis cerana cerana.

Honey bees play a critical role as pollinators. However, their reproduction success and survival face severe threats due to the deterioration of their living environment. Notably, environmental conditions during their preimaginal stage inside brood cells can influence their immune capabilities and overall health after emergence. During the in-cell developmental stage, workers are in close contact with cocoons, which can become a source of stress due to accumulated metals. To investigate this potential threat, experiments were conducted to examine the impact of cocoons in brood cells used to rear different generations on the metal content and detoxification gene expression levels in Apis cerana cerana. Our findings indicated significant differences in the layers, weight, base thickness, and metal contents like Cr, Cd, Pb, Mn, Ni, and As of cocoons in multi-generation brood cells compared to single-generation brood cells. These increases led to significant elevations in metal levels and upregulations of the four CYP450 detoxification genes in both six-day-old larvae and newly emerged workers. In conclusion, this study highlights the negative impact of cocoons in multi-generation brood cells on bee health and provides evidence supporting the development of rational apiculture management strategies for ecosystem stability.

Impact of Pollination on Pomegranate

Pollination mode affects crop quality and quantity in several economically significant crops. This study examined the effects of open pollination, Apis cerana pollination, hand, hand+ self-pollination, and self-pollination on pomegranate fruits. Open-pollinated plants had higher fruit length, weight, rind thickness, total soluble sugars, reducing sugar content, non-reducing sugar content, and total soluble solids, followed by A. cerana pollinated plants. A. cerana pollination increased pomegranate output and quality, including fruit shape, aril weight, TSS, and sugar content. The study found that open pollination and A. cerana integration increase pomegranate fruit yield and quality. A. cerana integration boosts farmers' income and conserves biodiversity.

Insulin receptor participates in the peripheral olfactory processes of honey bees (Apis cerana cerana).

Insulin receptors (InR) are an integral component of the insulin/insulin-like growth factor signaling pathway, which plays a vital role in insect development, lifespan, reproduction, and olfactory sensitivity. However, whether InR participate in the peripheral olfactory system of insects remains unclear. Recently, we found that 2-heptanone (2-HT) affects AcerInR expression, the gene for an InR protein, in Apis cerana cerana. We then examined the spatiotemporal expression profile of the gene in A.cerana cerana. The mRNA of AcerInR was primarily expressed in the antennae, wings, and legs of forager bees, which are probable chemosensory tissues. The results of fluorescence competitive binding assays, combined with site-directed mutagenesis, demonstrated that AcerOBP6 and AcerOBP14 exhibit strong binding affinities to 2-HT. Furthermore, after foragers were fed with double-stranded AcerInR, the expression levels of AcerOBP6 and AcerOBP14 decreased significantly, as did the electroantennogram responsiveness to 2-HT and some other odorants. In conclusion, our findings provide a foundation for understanding the involvement of AcerInR in the odor perception of A.cerana cerana. Moreover, they offer novel insights into the olfactory recognition mechanism in insects.

A case of Brugada phenocopy unmasked following giant Asian honey bee stings

This case report details a rare occurrence of Brugada phenocopy (type 1 Brugada electrocardiographic pattern) precipitated following giant Asian honey bee (Apis dorsata) stings in a previously healthy Sri Lankan male. The patient had an uneventful course. While similar ECG patterns can arise from non-genetic factors, Giant Asian honey bee sting as a causal factor is exceedingly unusual and unreported previously. This case adds to the spectrum of cardiac manifestations of Hymenopteran venom highlighting the requirement for close cardiac monitoring and clinical vigilance in these patients.

Spatial and Temporal Variations in Richness, Diversity and Abundance of Floral Visitors of Curry Plants (Bergera koenigii L.): Insights on Plant-Pollinator Interactions.

The reproductive success of flowering plants relates to flower-visitor communities and plant-pollinator interactions. These traits are species- and region-specific and vary across regions, pollinator groups, and plant species. However, little literature exists on the spatiotemporal variation in visitor activity, especially in India. Here, we aimed to depict the spatial and temporal variation in visitor activity on the curry plants (Bergera koenigii). Data were collected at different daytime slots from three vegetation zones (confirmed by field surveys and normalized difference vegetation index values in remote sensing)-dense, medium-density, and low-density vegetation in West Bengal, India. The visitors' richness, diversity, and abundance were higher in the area with dense vegetation. Considering daytime patterns, higher values for these parameters were obtained during 10.00-14.00 h. For most visitors, the flower handling time was shorter, and the visitation rate was higher in dense vegetation areas (at 10.00-14.00 h) than in medium- and low-density vegetation areas. The proportions of different foraging categories varied over time. Vital pollinators were Apis cerana, Apis dorsata, Appias libythea, Halictus acrocephalus, Nomia iridescens, and Tetragonula iridipennis. However, the effectiveness of pollinators remained region-specific. Therefore, it can be concluded that floral visitors' richness, diversity, abundance, and plant-visitor interactions varied spatially with their surrounding vegetation types and also changed daytime-wise.

Diversity of Pollinators on Sunflower

Sunflower crop was grown during rabi 2021 and summer 2022 at the M S Swaminathan School of Agriculture, Centurion University of Technology and Management, Gajapati, Odisha. Investigation has been made on diversity of different pollinating agents associated with sunflower by observing their abundance, diversity, percentage contribution following the statistical diversity indices. The present study revealed eighteen different pollinators associated with sunflower. The Indian honey bee, Apis cerana indica (F)was recorded as the most dominant hymenopteran pollinator followed by the rock bee Apis dorsata(F)from family Apidae. The foraging activity of A. c. indica was observed to be maximum during midflowering stage (1.72± 1.23 and 2.78± 1.81 bees/ capitulum/ min in 1st and 2nd season, respectively) withhighest activity during 01:00-02:00 PM (3.02 bees/ capitulum/min.) and 10:00 to 11:00 AM (4.42 bees/capitulum/ min) during first and second seasons respectively. The Apidae family members’ i.e. A. c. indica and A. dorsata have attained eudominant and other pollinators were belonging mostly to the recedentand subrecedent classes based on dominance status. The diversity indices showed mid flowering stageattracted the most diversified group of pollinators.

Host switch by honey bee parasitic mites leads to symbiont diversification

Host-parasite co-evolution is a reciprocal genetic change; however, the parasite may switch to a novel host, deviating from conventional co-evolution. Varroa destructor is a native parasite of the honey bee Apis cerana, and the mite has established infestation in another honey bee, Apis mellifera, causing colony failure. When mites switched to the novel host, they formed a distinct population from mites that remained on the native host. Consequently, this led to divergence in the microbiota associated with mites in two host populations. The microbes were conserved at the species level reflected by alpha diversity, with substantial relative abundance variance. Microbes found in mites were distinct from the bee microbiota. They mainly were pathogenic with antibiotic resistance, while a few bacterial taxa were previously found in honey bees, including Klebsiella pneumoniae and Pseudomanas aeruginosa. These symbionts may transfer between the mites and honey bees.

Combined pesticides in field doses weaken honey bee (Apis cerana F.) flight ability and analyses of transcriptomics and metabolomics

Imidacloprid, chlorpyrifos, and glyphosate rank among the most extensively employed pesticides worldwide. The effects of these pesticides and their combined on the flight capability of Apis cerana, and the potential underlying mechanisms remain uncertain. To investigate these effects, we carried out flight mill, transcriptome, and metabolome experiments. Our findings reveal that individual acute oral treatments with pesticides, specifically 20 μL of 10 ng/g imidacloprid (0.2 ng per bee), 30 ng/g chlorpyrifos (0.6 ng per bee), and 60 ng/g glyphosate (1.2 ng per bee), did not impact the flight capability of the bees. However, when bees were exposed to a combination of two or three pesticides, a notable reduction in flight duration and distance was observed. In the transcriptomic and metabolomic analyses, we identified 307 transcripts and 17 metabolites that exhibited differential expression following exposure to combined pesticides, primarily associated with metabolic pathways involved in energy regulation. Our results illuminate the intricate effects and potential hazards posed by combined pesticide exposures on bee behavior. These findings offer valuable insights into the synergistic potential of pesticide combinations and their capacity to impair bee behavior. Understanding these complex interactions is essential for comprehending the broader consequences of pesticide formulations on honey bee populations.

The use of drone congregation behaviour for population surveys of the honey bee Apis cerana

Honey bees (Apis spp.) are important pollinators in many natural and agro-ecosystems across the world. Effective means of surveying wild populations are therefore key to their conservation and management. One available survey method infers honey bee colony density from the genotype of drones (males) sampled from sites known as Drone Congregation Areas (DCAs). While this approach has been commonly used for the Western honey bee (A. mellifera), its feasibility for other Apis species is unknown. Here, we investigate drone congregation behaviour in the Asian honey bee Apis cerana in north-east Australia and its suitability for inferring colony density. As this A. cerana population is invasive, surveys in this case can aid in monitoring the population’s growth and spread. Over 5 years, we identified 30 DCAs, many of which were stable across time. DCAs were sheltered areas beside tree-lines or openings in the forest canopy. A. cerana drones joined DCAs during 1–2-h afternoon intervals and could be sampled at heights of 2–24 m via adhesive lines attached to helium balloons carrying lures coated in queen pheromone. Drones were more likely to be present at a DCA as temperature increased, though abiotic factors did not predict overall drone abundance. Drones could be sampled in low numbers even where colony density was extremely low. Based on the genotyping and inferred sibship of drones sampled at DCAs between 2016 and 2021, we estimate population density in Australia’s A. cerana to be in the range 1.1–8.1 colonies/km2. This extrapolates to a total population size in the range 11,000–83,000 colonies, with more refined estimates requiring better knowledge of drone flight distance and the effect of habitat on colony density. We conclude that population surveys based on drones from DCAs are possible for A. cerana and propose that this approach be part of a toolkit of methods used to monitor Asian honey bee populations in both their native and invasive ranges.

Glyphosate exposure affected longevity-related pathways and reduced survival in asian honey bees (Apis cerana)

Glyphosate (N-(phosphonomethyl)glycine, GLY) ranks among the most extensively used and effective herbicides globally. However, excessive GLY utilization poses a substantial threat to the survival of honey bees (Apis cerana). Here we monitored the survival status of A. cerana treated with GLY, and conducted transcriptome sequencing of the bee gut and head to further explore potential GLY influences at the molecular level. We observed that the mortality rate of bees increased as GLY concentration escalated. Pivotal pathways emerged in response to the GLY treatment, with a substantial number of differentially expressed genes enriched in the longevity regulating pathway - multiple species. This strongly suggested that GLY may influence the physiological behavior of bees by impacting this particular pathway. Moreover, our analysis revealed a notable reduction in the enzymatic activities of CYP450 and AChE in both the bee head and intestines of when exposed to GLY. Conversely, the enzymatic activity of superoxide dismutase (SOD) in the head remained unaffected, whereas in the intestines, it exhibited a significant increase. Additionally, prophenol oxidase (PPO) and glutathione-S-transferases (GSTs) displayed contrasting trends in enzymatic activity in both organs. This study offers valuable insights into how GLY impacted the survival of A. cerana.

Floral visitors of sesame (Sesamum indicum L.): Elucidating their nectar-robbing behaviour and impacts on the plant reproduction.

Nectar robbing is common in angiosperms, especially in long tubular flowers or flowers with spurs that keep nectar out of reach of visitors. However, the robbing behaviour of bees is less understood. Here, we studied the sesame visitors, their robbing behaviour, and the impacts of robbing on plant reproductive fitness. Diverse insect species (primarily members of Hymenoptera) visited sesame flowers. The most effective pollinators were Amegilla zonata, Apis cerana, Apis dorsata, Apis florea, Ceratina binghami, Halictus acrocephalus and Xylocopa amethystina. Almost all visitors with variable percentages revealed the nectar-robbing phenomenon. Robbing activity depended on a complex of multiple attributes, including the visitor's body size, the corolla tube length, the availability and accessibility of nectar, and the resource-collecting task allocation of bees. Robbing activity varied according to flower-visiting species, flowering period and daytime. Robbing was comparatively higher in the late flowering period at 10.00-14.00 h. In the case of robbing visits, flower handling time was lower, and the visitation rate remained higher than non-robbing visits. Robbing visits did not significantly affect fruit and seed sets of sesame. Therefore, we can interpret the nectar-robbing interactions on sesame as commensal, with pollinators benefitting without altering the plant's reproductive fitness.

The Participating Role of Honeybee Farming Groups (Apis cerana) in Kamangta Village, Tombulu Subdistrict, Minahasa Regency, North Sulawesi

The Participating Role of Honeybee Farming Groups (Apis cerana) in Kamangta Village, Tombulu Subdistrict, Minahasa Regency, North Sulawesi

The effects of meliponicultural use of Tetragonula laeviceps on other bee pollinators and pollination efficacy of lemon.

The augmentation of pollination success in lemon (Citrus limon Eureka) flowers remains contingent on the involvement of bee pollinators. With wild bee pollinator populations declining in agroecosystems, meliponiculture has emerged as a potential option in Indonesia. This study aimed to investigate the effects of meliponicultural use of Tetragonula laeviceps on diversity, foraging behavior, and monthly population of bee pollinators, as well as lemon pollination efficacy with and without meliponiculture treatment during two periods. Using scan and focal sampling methods in first and second periods, the study found that the diversity of wild bee pollinators was six species (Apis cerana, Lasioglossum albescens, Megachile laticeps, Xylocopa confusa, Xylocopa latipes, and Xylocopa caerulea), and T. laeviceps when using meliponiculture. The relative abundance and daily foraging activity of wild bee pollinators were initially reduced in the first period (March-June) and then maintained in the second period (July-October). T. laeviceps foraged on the flowers, involving specific sequences for 72 s with highest visitation rate of 0.25 flowers/h from 10:00-13:00. Light intensity was observed to be the most influential factor for bee pollinator density. Pollination efficacy results showed that meliponiculture usage has greater benefit compared to meliponiculture absence across various parameters, including fruit sets, fruit weight, yield, and estimated productivity. The effects of meliponicultural use of T. laeviceps can enhance lemon pollination efficacy while preserving the diversity of wild insect pollinators. This suggests that meliponiculture stingless bees could be a beneficial practice in agroecosystems, especially in tropical regions where wild bee populations and diversity are declining.