Culturable yeast community associated with grape must and honey bees sampled from apiaries located in the vineyards.

AIM: In this study, we investigated culturable yeast community, present in grape must sampled from vineyards with apiaries on the borders, and in honey bees collected in these apiaries. METHODS AND RESULTS: To this aim, yeasts isolated from spontaneously fermented grapes randomly collected in two vineyards (P1 and P2) with apiaries on the borders (A1 and A2) were compared to those isolated from spontaneously fermented grapes collected from a vineyard without apiary (P4). At the same time, yeast community was analyzed on bees collected in each apiary placed in the vineyards, in comparison to yeasts isolated from an apiary (A3) located far from the vineyards. The analysis was performed for two consecutive years (2021 and 2022). The isolated yeasts were identified by restriction analysis of amplified ITS region, followed by sequencing of ITS fragment. RESULTS: Our research showed that the presence of apiaries seems to increase yeast counts of grape must, in particular of Saccharomyces cerevisiae; furthermore, the permanence of apiaries in the vineyards allowed the recovering of these yeasts also from bees. CONCLUSIONS: Our findings seem to corroborate the role of bees as vectors and reservoir of oenologically relevant yeasts, such as a source of non-conventional yeasts with potential biotechnological applications.

Effects of chronic oral exposure to insecticide teflubenzuron on the midgut of the honey bee Apis mellifera workers: histopathological insights into pesticide toxicity.

The honey bee Apis mellifera plays a significant role as a pollinator of native and cultivated plants, by increasing the productivity of several cultures, preserving the flora, and producing forest seeds. However, bee populations are declining worldwide, including A. mellifera, due to Colony Collapse Disorder, mainly resulting from the constant use of pesticides in the crops. Teflubenzuron is a physiological insecticide that belongs to the benzoylurea group, which inhibits chitin synthesis, the main component of the insect integument classified as safe for non-target insects, including bees. However, its effect on non-target organs of insects remains unknown. The midgut is the main organ of the digestive tract, which works in digestion and absorption and may be exposed to pesticides that contaminate food resources. The present work aimed to verify if the insecticide teflubenzuron is toxic and has histopathological effects on the midgut of A. mellifera adult workers. Workers exposed orally and chronically to the field-realistic concentration of teflubenzuron present 81.54% mortality. The epithelium of the midgut of these bees presents high vacuolization, spherocrystals, cell fragments released to the organ lumen, apocrine secretion, nuclear pyknosis, loss of cell-cell contact, and damage to regenerative cell nests and to the peritrophic matrix. These results indicate that the chitin synthesis-inhibiting insecticide teflubenzuron is toxic to A. mellifera after chronic oral exposure, at realistic field concentration, although it is classified as non-toxic to adult and non-target insects.

Potential exposure of honey bees to neonicotinoid seed treatments in US rice.

Neonicotinoid insecticide seed treatments are commonly used in rice (Oryza sativa) production to control rice water weevil (Lisorhoptrus oryzophilus). With the use of neonicotinoid seed treatments, there is potential that honey bees (Apis mellifera) could be exposed to neonicotinoids through translocation to the pollen. Studies were conducted in 2015 and 2016 to determine the level of neonicotinoids present in flag leaves, pollen, and grain of rice. Thiamethoxam was applied as a seed treatment and foliar prior to flooding. Clothianidin was applied as a seed treatment and as a foliar at a preflood and postflood timing. Subsamples of flag leaves, pollen, and grain were analyzed for positive neonicotinoid detections and abundance. Thiamethoxam was detected in 8.9% of samples and clothianidin was detected in 1.4% of samples. For both thiamethoxam and clothianidin, more positive samples were observed in flag leaf samples than in pollen or grain. An average of 4.30 ng/g of thiamethoxam was detected in flag leaves from seed-applied thiamethoxam. An average of 1.25 ng/g of clothianidin was found in flag leaves from a preflood application of clothianidin. A survey of honey bees present in rice fields was conducted in Mississippi and Arkansas to determine the abundance of honey bees present in rice fields based on the time of day. Honey bee densities were low in rice, with less than 5% and 3% positive detections observed in Mississippi and Arkansas, respectively. More positive detections and higher densities of honey bees were observed for mid-day sampling than for morning or evening sampling.

Effects of floral resources on honey bee populations in Mexico: Using dietary metabarcoding to examine landscape quality in agroecosystems.

The decline of honey bee populations significantly impacts the human food supply due to poor pollination and yield decreases of essential crop species. Given the reduction of pollinators, research into critical landscape components, such as floral resource availability and land use change, might provide valuable information about the nutritional status and health of honey bee colonies. To address this issue, we examine the effects of landscape factors like agricultural area, urban area, and climatic factors, including maximum temperature, minimum temperature, relative humidity, and precipitation, on honey bee hive populations and nutritional health of 326 honey bee colonies across varying landscapes in Mexico. DNA metabarcoding facilitated the precise identification of pollen from 267 plant species, encompassing 243 genera and 80 families, revealing a primary herb-based diet. Areas characterized by high landscape diversity exhibited greater pollen diversity within the colony. Conversely, colonies situated in regions with higher proportions of agricultural and urban landscapes demonstrated lower bee density. The maximum ambient temperature outside hives positively correlated with pollen diversity, aligning with a simultaneous decrease in bee density. Conversely, higher relative humidity positively influenced both the bee density of the colony and the diversity of foraged pollen. Our national-level study investigated pollen dietary availability and colony size in different habitat types, latitudes, climatic conditions, and varied levels and types of disturbances. This effort was taken to gain a better insight into the mechanisms driving declines in honey bee populations. This study illustrates the need for more biodiverse agricultural landscapes, the preservation of diverse habitats, and the conservation of natural and semi-natural spaces. These measures can help to improve the habitat quality of other bee species, as well as restore essential ecosystem processes, such as pollination and pest control.

What proteomics has taught us about honey bee (Apis mellifera) health and disease.

The Western honey bee, Apis mellifera, is currently navigating a gauntlet of environmental pressures, including the persistent threat of parasites, pathogens, and climate change - all of which compromise the vitality of honey bee colonies. The repercussions of their declining health extend beyond the immediate concerns of apiarists, potentially imposing economic burdens on society through diminished agricultural productivity. Hence, there is an imperative to devise innovative monitoring techniques for assessing the health of honey bee populations. Proteomics, recognized for its proficiency in biomarker identification and protein-protein interactions, is poised to play a pivotal role in this regard. It offers a promising avenue for monitoring and enhancing the resilience of honey bee colonies, thereby contributing to the stability of global food supplies. This review delves into the recent proteomic studies of A. mellifera, highlighting specific proteins of interest and envisioning the potential of proteomics to improve sustainable beekeeping practices amidst the challenges of a changing planet.

Are man-made hives valid thermal surrogates for natural honey bee nests (Apismellifera)?

Honey bees preferentially occupy thick walled tall narrow tree cavities and attach their combs directly to the nest wall, leaving periodic gaps. However, academic research and beekeeping are conducted in squat, thin walled man made hives, with a continuous gap between the combs and the walls and roof. Utilising a computational fluid dynamics (CFD) model of thermoregulating bees in complete nests in trees and thin walled man made hives, with the average size of tree comb gaps determined from honey bee occupied synthetic tree nests, this research compared the metabolic energy impacts of comb gaps and vertical movement of the thermoregulated brood area. This shows their heat transfer regimes are disparate, including: bee space above combs increases heat loss by up to ∼70%; hives, compared to tree nests, require at least 150% the density of honey bees to arrest convection across the brood area. Tree cavities have a larger vertical freedom, a greater thermal resistance and can make dense clustering redundant. With the thermal environment being critical to honey bees, the magnitude and scope of these differences suggest that some hive based behavioural research needs extra validation to be considered non-anthropogenic, and some bee keeping practices are sub-optimal.

The Influence of Environmental Factors on Site Selection Augment Breeding Success in Honey Bees: An Insight of Honey Bee Genetic Resource Conservation.

Honey bee reproductive behavior involves a complicated mating system that embodies a number of factors, including environmental and human-induced factors. Controlled breeding in isolated mating stations is a prerequisite to maintain the genetic resources of honey bees through natural mating. The concept of controlled mating is a challenge in most beekeeping operations due to its low mating success rate. Therefore, a detailed investigation into the suitability of isolated mating stations is of interest. Thus, we bred two subspecies of honey bees (Apis cerana koreana and Apis mellifera L.) in isolated mating stations (island) from 2021 to 2023 and in an open breeding station in 2023. Our results demonstrate that the highest percentage of the mating success rate in isolated mating stations was recorded in the Wido Island, which had the highest percentage of bare land, coniferous forests, deciduous forests, fields, and mixed forests. The mating success rate was higher in the summer and spring for A. cerana and A. mellifera, respectively. The mating success rate was higher in open mating compared to controlled mating (Island) and did not vary between pure-breeding and cross-breeding lines. Our findings suggested that mating stations with mixed forest and fields are potential sites for the successful breeding of honey bees.

Higher prevalence of sacbrood virus in Apis mellifera (Hymenoptera: Apidae) colonies after pollinating highbush blueberries.

Highbush blueberry pollination depends on managed honey bees (Apis mellifera) L. for adequate fruit sets; however, beekeepers have raised concerns about the poor health of colonies after pollinating this crop. Postulated causes include agrochemical exposure, nutritional deficits, and interactions with parasites and pathogens, particularly Melisococcus plutonius [(ex. White) Bailey and Collins, Lactobacillales: Enterococcaceae], the causal agent of European foulbrood disease, but other pathogens could be involved. To broadly investigate common honey bee pathogens in relation to blueberry pollination, we sampled adult honey bees from colonies at time points corresponding to before (t1), during (t2), at the end (t3), and after (t4) highbush blueberry pollination in British Columbia, Canada, across 2 years (2020 and 2021). Nine viruses, as well as M. plutonius, Vairimorpha ceranae, and V. apis [Tokarev et al., Microsporidia: Nosematidae; formerly Nosema ceranae (Fries et al.) and N. apis (Zander)], were detected by PCR and compared among colonies located near and far from blueberry fields. We found a significant interactive effect of time and blueberry proximity on the multivariate pathogen community, mainly due to differences at t4 (corresponding to ~6 wk after the beginning of the pollination period). Post hoc comparisons of pathogens in near and far groups at t4 showed that detections of sacbrood virus (SBV), which was significantly higher in the near group, not M. plutonius, was the primary driver. Further research is needed to determine if the association of SBV with highbush blueberry pollination is contributing to the health decline that beekeepers observe after pollinating this crop.

Occasional and constant exposure to dietary ethanol shortens the lifespan of worker honey bees.

Honey bees (Apis mellifera) are one of the most crucial pollinators, providing vital ecosystem services. Their development and functioning depend on essential nutrients and substances found in the environment. While collecting nectar as a vital carbohydrate source, bees routinely encounter low doses of ethanol from yeast fermentation. Yet, the effects of repeated ethanol exposure on bees' survival and physiology remain poorly understood. Here, we investigate the impacts of constant and occasional consumption of food spiked with 1% ethanol on honey bee mortality and alcohol dehydrogenase (ADH) activity. This ethanol concentration might be tentatively judged close to that in natural conditions. We conducted an experiment in which bees were exposed to three types of long-term diets: constant sugar solution (control group that simulated conditions of no access to ethanol), sugar solution spiked with ethanol every third day (that simulated occasional, infrequent exposure to ethanol) and daily ethanol consumption (simulating constant, routine exposure to ethanol). The results revealed that both constant and occasional ethanol consumption increased the mortality of bees, but only after several days. These mortality rates rose with the frequency of ethanol intake. The ADH activity remained similar in bees from all groups. Our findings indicate that exposure of bees to ethanol carries harmful effects that accumulate over time. Further research is needed to pinpoint the exact ethanol doses ingested with food and exposure frequency in bees in natural conditions.

Tropical and montane Apis cerana show distinct dance-distance calibration curves.

Social bees have evolved sophisticated communication systems to recruit nestmates to newly found food sources. As foraging ranges can vary from a few hundred meters to several kilometers depending on the environment or season, populations of social bee species living in different climate zones likely show specific adaptations in their recruitment communication. Accordingly, studies in the western honey bee, Apis mellifera, demonstrated that temperate populations exhibit shallower dance-calibration curves compared with tropical populations. Here, we report the first comparison of calibration curves for three Indian Apis cerana lineages: the tropical Apis indica, and the two montane Himalayan populations Apis cerana cerana (Himachal Pradesh) and Apis cerana kashmirensis (Jammu and Kashmir). We found that the colonies of the two montane A. cerana populations show dance-distance calibration curves with significantly shallower slopes than those of the tropical A. indica. Next, we transferred A. c. cerana colonies to Bangalore (∼ 2600 km away) to obtain calibration curves in the same location as A. indica. The common garden experiment confirmed this difference in slopes, implying that the lineages exhibit genetically fixed differences in dance-distance coding. However, the slopes of the calibration curves of the transferred A. c. cerana colonies were also significantly higher than those of the colonies tested in their original habitat, indicating an important effect of the environment. The differences in dance-distance coding between temperate and tropical A. cerana lineages resemble those described for Apis mellifera, suggesting that populations of both species independently evolved similar adaptations.

Dominance of African racial ancestry in honey bee colonies of Mexico 30 years after the migration of hybrids from South America.

The Africanized honey bee, a hybrid of Apis mellifera scutellata from Africa with European subspecies, has been considered an invasive species and a problem for beekeeping. Africanized bees arrived in Mexico in 1986, 30 years after their accidental release in Brazil. Although government programs were implemented for its eradication, Africanized populations persist in Mexico, but precise information on the patterns of genetic introgression and racial ancestry is scarce. We determined maternal and parental racial ancestry of managed and feral honey bees across the five beekeeping regions of Mexico, using mitochondrial (mtDNA, COI-COII intergenic region) and nuclear markers (94 ancestrally informative SNPs), to assess the relationship between beekeeping management, beekeeping region, altitude, and latitude with the distribution of maternal and parental racial ancestry. Results revealed a predominantly African ancestry in the Mexican honey bees, but the proportion varied according to management, beekeeping regions, and latitude. The Mexican honey bees showed 31 haplotypes of four evolutionary lineages (A, M, C, and O). Managed honey bees had mitochondrial and nuclear higher proportions of European ancestry than feral honey bees, which had a higher proportion of African ancestry. Beekeeping regions of lower latitudes had higher proportions of African nuclear ancestry. Managed and feral honey bees showed differences in the proportion of maternal and nuclear racial ancestry. Managed honey bees from the Yucatan Peninsula and feral honey bees had a higher mtDNA than nuclear proportions of African ancestry. Managed honey bees, except those on the Yucatan Peninsula, had a higher nuclear than mtDNA proportion of African ancestry. Our study demonstrates that Africanized honey bee populations are genetically diverse and well established in Mexico, which highlights the limitations of management and government programs to contain the Africanization process and demands the incorporation of this lineage in any breeding program for sustainable beekeeping.

Measuring honey bee feeding rhythms with the BeeBox, a platform for nectar foraging insects.

In honey bees, most studies of circadian rhythms involve a locomotion test performed in a small tube, a tunnel, or at the hive entrance. However, despite feeding playing an important role in honey bee health or fitness, no demonstration of circadian rhythm on feeding has been performed until recently. Here, we present the BeeBox, a new laboratory platform for bees based on the concept of the Skinner box, which dispenses discrete controlled amounts of food (sucrose syrup) following entrance into an artificial flower. We compared caged groups of bees in 12 h-12 h light/dark cycles, constant darkness and constant light and measured average hourly syrup consumption per living bee. Food intake was higher in constant light and lower in constant darkness; mortality increased in constant light. We observed rhythmic consumption with a period longer than 24 h; this is maintained in darkness without environmental cues, but is damped in the constant light condition. The BeeBox offers many new research perspectives and numerous potential applications in the study of nectar foraging animals.

Can Bees Detect Perfluorooctane Sulfonate (PFOS)?

The European honey bee (Apis mellifera) is an important crop pollinator threatened by multiple stressors, including exposure to contaminants. Perfluorooctane sulfonate (PFOS) is a persistent global contaminant that accumulates and biomagnifies in food chains and is detected in honey. Even sublethal exposure to PFOS is detrimental to bee health, but exposure routes are unclear and nothing is known about bee response (detection, avoidance, or attraction) to PFOS. Using Y-mazes, we studied the response of individual bees to PFOS-spiked sugar syrup at four concentrations, 0.02, 30, 61 and 103 µg L-1. Bee activity, choice behavior, and drink duration for unspiked and spiked sugar syrup was recorded for 10 min in the Y-maze system. Most bees (≥80%) tasted and then drank the sugar syrup solutions, including the PFOS-contaminated syrup. Only at 61 and 103 µg L-1 did bees significantly avoid drinking PFOS-spiked syrup, and only when given a choice with unspiked syrup. When the choice of consuming unspiked syrup was removed, the bees drank PFOS-spiked syrup at all the PFOS concentrations tested, and avoidance was not evident. Avoidance was not observed in any treatment at 0.02 or 30 µg L-1 PFOS, concentrations that are frequently reported in environmental waters in contaminated areas. These findings confirm that bees will access PFOS-contaminated resources at concentrations detrimental to the colony health, and provide evidence of potential exposure pathways that may threaten crop pollination services and also human health via food chain transfer in PFOS-contaminated areas. Environ Toxicol Chem 2024;43:1638-1647. © 2024 SETAC.

Droplet Digital RT-PCR (dd RT-PCR) Detection of SARS-CoV-2 in Honey Bees and Honey Collected in Apiaries across the Campania Region.

Coronaviruses (CoVs), a subfamily of Orthocoronavirinae, are viruses that sometimes present a zoonotic character. Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is responsible for the recent outbreak of COVID-19, which, since its outbreak in 2019, has caused about 774,593,066 confirmed cases and 7,028,881 deaths. Aereosols are the main route of transmission among people; however, viral droplets can contaminate surfaces and fomites as well as particulate matter (PM) in suspensions of natural and human origin. Honey bees are well known bioindicators of the presence of pollutants and PMs in the environment as they can collect a great variety of substances during their foraging activities. The aim of this study was to evaluate the possible role of honey bees as bioindicators of the prevalence SARS-CoV-2. In this regard, 91 samples of honey bees and 6 of honey were collected from different apiaries of Campania region (Southern Italy) in four time periods from September 2020 to June 2022 and were analyzed with Droplet Digital RT-PCR for SARS-CoV-2 target genes Orf1b and N. The screening revealed the presence of SARS-CoV-2 in 12/91 in honey bee samples and in 2/6 honey samples. These results suggest that honey bees could also be used as indicators of outbreaks of airborne pathogens such as SARS-CoV-2.

Use of Honey Bees and Hive Products as Bioindicators to Assess Environmental Contamination in Targeted Areas of the Campania Region (Italy).

In recent years, biomonitoring has gained more attention, particularly when assessing the environmental health of significant areas, such as those near waste-to-energy facilities. These requirements coincide with the chance to detect environmental pollutants using sensitive organisms. Bees were shown to be quite effective in evaluating the presence of certain compounds by analyzing their associated matrices, such as pollen, honey, or wax. In our study, we employed the honey bee (Apis mellifera) as an indicator to initially monitor the vicinity of the waste-to-energy plant in Acerra, which is situated in the Campania region of Italy. The primary aim was to determine whether the facility was accountable for any environmental releases of dioxins or dioxin-like compounds. Then, we assessed the presence of additional pollutants in the same area, including trace elements, polycyclic aromatic hydrocarbons, and pesticides, released by human activities. To obtain further information about environmental quality, a second biomonitoring station was installed near the Caivano S.T.I.R. (Waste Shredding, Sifting, and Packaging Plant). The results showed the dioxin levels did not exceed predetermined limitations at the Acerra site, thus demonstrating the efficacy of the waste-to-energy facility and the bees' ability to detect the presence of other pollutants. Additionally, this biomonitoring system exhibited sensitivity to environmental variations, thereby enabling the evaluation of xenobiotic flux between two proximate zones and across temporal scales. This pioneering study suggests the advantages of utilizing bees to detect a wide range of contaminants, thereby providing valuable insights into environmental quality and potential health risks for both ecosystems and human populations.

Uncovering hidden dangers: The combined toxicity of abamectin and lambda-cyhalothrin on honey bees.

Studying the toxic effects of pesticides on bees has consistently been a prominent area of interest for researchers. Nonetheless, existing research has predominantly concentrated on individual toxicity assessments, leaving a gap in our understanding of mixed toxicity. This study delves into the individual and combined toxic effects of abamectin (ABA) and lambda-cyhalothrin (LCY) on honey bees (Apis mellifera) in laboratory settings. We discovered that ABA (96 h-LC50 value of 0.079 mg/L) exhibited greater acute toxicity to honey bees compared to LCY (96 h-LC50 value of 9.177 mg/L). Moreover, the mixture of ABA and LCY presented an acute antagonistic effect on honey bees. Additionally, our results indicated that exposure to LCY, at medium concentration, led to a reduction in the abundance of gut core bacterium Snodgrassella. However, an increase in the abundance of Bifidobacterium was noted when exposed to a medium concentration of LCY and its mixture with ABA. Transcriptomic analysis revealed significant regulation of certain genes in the medium concentration of all three treatments compared to the control group, primarily enriching in metabolism and immune-related pathways. Following chronic exposure to field-relevant concentrations of ABA, LCY, and their mixture, there were significant alterations in the activities of immunity-related enzyme polyphenol oxidase (PPO) and detoxification enzymes glutathione S-transferase (GST) and carboxylesterase (CarE). Additionally, the expression of four genes (abaecin, cyp9e2, cyp302a1, and GstD1) associated with immune and detoxification metabolism was significantly altered. These findings suggest a potential health risk posed by the insecticides ABA and LCY to honey bees. Despite exhibiting acute antagonistic effect, mixed exposure still induced damage to bees at all levels. This study advances our knowledge of the potential adverse effects of individual or combined exposure to these two pesticides on non-target pollinators and offers crucial guidance for the use of insecticides in agricultural production.

Quantifying Abdominal Coloration of Worker Honey Bees.

The main drawback in using coloration to identify honey bee subspecies is the lack of knowledge regarding genetic background, subjectivity of coloration grading, and the effect of the environment. The aim of our study was to evaluate the effect of environmental temperature on the abdominal coloration of honey bee workers and to develop a tool for quantifying abdominal coloration. We obtained four frames of honey bee brood from two colonies and incubated them at two different temperatures (30 and 34 °C). One colony had workers exhibiting yellow marks on the abdomen, while the other did not. We collected hatched workers and photographed abdomens. Images were analyzed using custom-written R script to obtain vectors that summarize the coloration over the abdomen length in a single value-coloration index. We used UMAP to reduce the dimensions of the vectors and to develop a classification procedure with the support vector machine method. We tested the effect of brood origin and temperature on coloration index with ANOVA. UMAP did not distinguish individual abdomens according to experimental group. The trained classifier sufficiently separated abdomens incubated at different temperatures. We improved the performance by preprocessing data with UMAP. The differences among the mean coloration index values were not significant between the gray groups incubated at different temperatures nor between the yellow groups. However, the differences between the gray and yellow groups were significant, permitting options for application of our tool and the newly developed coloration index. Our results indicate that the environmental temperature in the selected range during development does not seem to impact honey bee coloration significantly. The developed color-recording protocol and statistical analysis provide useful tools for quantifying abdominal coloration in honey bees.

Unravelling the impact of climate change on honey bees: An ensemble modelling approach to predict shifts in habitat suitability in Queensland, Australia.

Honey bees play a vital role in providing essential ecosystem services and contributing to global agriculture. However, the potential effect of climate change on honey bee distribution is still not well understood. This study aims to identify the most influential bioclimatic and environmental variables, assess their impact on honey bee distribution, and predict future distribution. An ensemble modelling approach using the biomod2 package in R was employed to develop three models: a climate-only model, an environment-only model, and a combined climate and environment model. By utilising bioclimatic data (radiation of the wettest and driest quarters and temperature seasonality) from 1990 to 2009, combined with observed honey bee presence and pseudo absence data, this model predicted suitable locations for honey bee apiaries for two future time spans: 2020-2039 and 2060-2079. The climate-only model exhibited a true skill statistic (TSS) value of 0.85, underscoring the pivotal role of radiation and temperature seasonality in shaping honey bee distribution. The environment-only model, incorporating proximity to floral resources, foliage projective cover, and elevation, demonstrated strong predictive performance, with a TSS of 0.88, emphasising the significance of environmental variables in determining habitat suitability for honey bees. The combined model had a higher TSS of 0.96, indicating that the combination of climate and environmental variables enhances the model's performance. By the 2020-2039 period, approximately 88% of highly suitable habitats for honey bees are projected to transition from their current state to become moderate (14.84%) to marginally suitable (13.46%) areas. Predictions for the 2060-2079 period reveal a concerning trend: 100% of highly suitable land transitions into moderately (0.54%), marginally (17.56%), or not suitable areas (81.9%) for honey bees. These results emphasise the critical need for targeted conservation efforts and the implementation of policies aimed at safeguarding honey bees and the vital apiary industry.

The Effects of Artificial Diets on the Expression of Molecular Marker Genes Related to Honey Bee Health.

Honey bees are commonly used to study metabolic processes, yet the molecular mechanisms underlying nutrient transformation, particularly proteins and their effects on development, health, and diseases, still evoke varying opinions among researchers. To address this gap, we investigated the digestibility and transformation of water-soluble proteins from four artificial diets in long-lived honey bee populations (Apis mellifera ligustica), alongside their impact on metabolism and DWV relative expression ratio, using transcriptomic and protein quantification methods. Diet 2, characterized by its high protein content and digestibility, was selected for further analysis from the other studied diets. Subsequently, machine learning was employed to identify six diet-related molecular markers: SOD1, Trxr1, defensin2, JHAMT, TOR1, and vg. The expression levels of these markers were found to resemble those of honey bees who were fed with Diet 2 and bee bread, renowned as the best natural food. Notably, honey bees exhibiting chalkbrood symptoms (Control-N) responded differently to the diet, underscoring the unique nutritional effects on health-deficient bees. Additionally, we proposed a molecular model to elucidate the transition of long-lived honey bees from diapause to development, induced by nutrition. These findings carry implications for nutritional research and beekeeping, underscoring the vital role of honey bees in agriculture.

Influence of Age of Infection on the Gut Microbiota in Worker Honey Bees (Apis mellifera iberiensis) Experimentally Infected with Nosema ceranae.

The gut microbiota of honey bees has received increasing interest in the past decades due to its crucial role in their health, and can be disrupted by pathogen infection. Nosema ceranae is an intracellular parasite that affects the epithelial cells of the midgut, altering gut homeostasis and representing a major threat to honey bees. Previous studies indicated that younger worker bees are more susceptible to experimental infection by this parasite, although the impact of infection and of age on the gut bacterial communities remains unclear. To address this, honey bees were experimentally infected with a consistent number of N. ceranae spores at various ages post-emergence (p.e.) and the gut bacteria 7 days post-infection (p.i.) were analysed using real-time quantitative PCR, with the results compared to non-infected controls. Infected bees had a significantly higher proportion and load of Gilliamella apicola. In respect to the age of infection, the bees infected just after emergence had elevated loads of G. apicola, Bifidobacterium asteroides, Bombilactobacillus spp., Lactobacillus spp., Bartonella apis, and Bombella apis. Moreover, the G. apicola load was higher in bees infected at nearly all ages, whereas older non-infected bees had higher loads of Bifidobacterium asteroides, Bombilactobacillus spp., Lactobacillus spp., Ba. apis, and Bo apis. These findings suggest that N. ceranae infection and, in particular, the age of bees at infection modulate the gut bacterial community, with G. apicola being the most severely affected species.