Microbial ensemble in the hives: deciphering the intricate gut ecosystem of hive and forager bees of Apis mellifera.

BACKGROUND: The gut microbiome of honey bees significantly influences vital traits and metabolic processes, including digestion, detoxification, nutrient provision, development, and immunity. However, there is a limited information is available on the gut bacterial diversity of western honey bee populations in India. This study addresses the critical knowledge gap and outcome of which would benefit the beekeepers in India. METHODS AND RESULTS: This study investigates the gut bacterial diversity in forager and hive bees of Indian Apis mellifera, employing both culture-based and culture-independent methods. In the culturable study, a distinct difference in gut bacterial alpha and beta diversity between forager and hive bees emerges. Firmicutes, Proteobacteria, and Actinobacteria dominate, with hive bees exhibiting a Firmicutes-rich gut (65%), while foragers showcase a higher proportion of Proteobacteria (37%). Lactobacillus in the hive bee foregut aligns with the findings by other researchers. Bacterial amplicon sequencing analysisreveals a more intricate bacterial composition with 18 identified phyla, expanding our understanding compared to culturable methods. Hive bees exhibit higher community richness and diversity, likely due to diverse diets and increased social interactions. The core microbiota includes Snodgrassella alvi, Gilliamella apicola, and Bombilactobacillus mellis and Lactobacillus helsingborgensis, crucial for digestion, metabolism, and pathogen resistance. The study emphasises bacteria's role in pollen and nectar digestion, with specific groups like Lactobacillus and Bifidobobacterium spp. associated with carbohydrate metabolism and polysaccharide breakdown. These microbes aid in starch and sucrose digestion, releasing beneficial short-chain fatty acids. CONCLUSION: This research highlights the intricate relationship between honey bees and their gut microbiota, showcasing how the diverse and complex microbiome helps bees overcome dietary challenges and enhances overall host health. Understanding these interactions contributes to bee ecology knowledge and has implications for honey bee health management, emphasising the need for further exploration and conservation efforts.

Contaminant dynamics in honey bees and hive products of apiaries from environmentally contrasting Argentinean regions.

Argentina is a leading honey producer and honey bees are also critical for pollination services and wild plants. At the same time, it is a major crop producer with significant use of insecticides, posing risks to bees. Therefore, the presence of the highly toxic insecticide chlorpyrifos, and forbidden contaminants (organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs)) was investigated in honey bee, beebread, wax and honey samples in apiaries from three contrasting regions of Argentina. Chlorpyrifos was detected in all samples with higher levels during period 1 (spring) in contrast to period 2 (fall), agreeing with its season-wise use in different crops, reaching 3.05 ng/g in honey bees. A subsequent first-tier pesticide hazard analysis revealed that it was relevant to honey bee health, mainly due to the high concentrations found in wax samples from two sites, reaching 132.4 ng/g. In addition, wax was found to be the most contaminated matrix with a prevalence of OCPs (∑OCPs 58.23-172.99 ng/g). Beebread samples showed the highest concentrations and diversity of pesticide residues during period 1 (higher temperatures). A predominance of the endosulfan group was registered in most samples, consistent with its intensive past use, especially in Central Patagonia before its prohibition. Among the industrial compounds, lighter PCB congeners dominated, suggesting the importance of atmospheric transport. The spatio-temporal distribution of pesticides shows a congruence with the environmental characteristics of the areas where the fields are located (i.e., land use, type of productive activities and climatic conditions). Sustained monitoring of different pollutants in beekeeping matrices is recommended to characterize chemical risks, assess the health status of honey bee hives and the pollution levels of different agroecosystems. This knowledge will set a precedent for South America and be helpful for actions focused on the conservation of pollination services, apiculture and ecosystems in Argentina.

Shotgun Metagenomics Reveals Minor Micro"bee"omes Diversity Defining Differences between Larvae and Pupae Brood Combs.

Bees represent not only a valuable asset in agriculture, but also serve as a model organism within contemporary microbiology. The metagenomic composition of the bee superorganism has been substantially characterized. Nevertheless, traditional cultural methods served as the approach to studying brood combs in the past. Indeed, the comb microbiome may contribute to determining larval caste differentiation and hive immunity. To further this understanding, we conducted a shotgun sequencing analysis of the brood comb microbiome. While we found certain similarities regarding species diversity, it exhibits significant differentiation from all previously described hive metagenomes. Many microbiome members maintain a relatively constant ratio, yet taxa with the highest abundance level tend to be ephemeral. More than 90% of classified metagenomes were Gammaproteobacteria, Bacilli and Actinobacteria genetic signatures. Jaccard dissimilarity between samples based on bacteria genus classifications hesitate from 0.63 to 0.77, which for shotgun sequencing indicates a high consistency in bacterial composition. Concurrently, we identified antagonistic relationships between certain bacterial clusters. The presence of genes related to antibiotic synthesis and antibiotic resistance suggests potential mechanisms underlying the stability of comb microbiomes. Differences between pupal and larval combs emerge in the total metagenome, while taxa with the highest abundance remained consistent. All this suggests that a key role in the functioning of the comb microbiome is played by minor biodiversity, the function of which remains to be established experimentally.

Draft genomes of Paenibacillus larvae isolated from honeybee colonies (Apis mellifera) in Fiji.

Here, we report draft genomic sequences from three Paenibacillus larvae isolates, the causative agent of American Foulbrood (AFB), obtained from honeybee colonies of Apis mellifera in Fiji, which allow both enterobacterial repetitive intergenic consensus and multilocus sequence typing genotypes to be elucidated for Fijian AFB.

Hive Orientation and Colony Strength Affect Honey Bee Colony Activity during Almond Pollination.

The foraging activity of honey bees used to pollinate almonds was examined in relation to their hive entrance orientation and colony strength. Twenty-four colonies of honey bees, twelve in each group, were situated with their entrances facing east and west cardinal points. Bee out counts were recorded continuously and hive weight data at ∼10 min intervals from 17 February to 15 March 2023. Colony strength was assessed using the frames of adult bees (FOB) metric. East-facing hives started flight 44.2 min earlier than west-facing hives. The hive direction did not affect the timing of the cessation of foraging activity. The hive strength played a significant role: hives assessed as weak (≤3.0 FOB) commenced foraging activity 45 min later than strong hives (>3.0 FOB) and ceased foraging activity 38.3 min earlier. Hive weight data did not detect effects of either the hive direction or colony strength on the commencement and cessation of foraging activity, as determined using piecewise regression on 24 h datasets. However, the hive weight loss due to foraging activity at the start of foraging activity was significantly affected by both direction (East > West) and colony strength (Strong > Weak). Our study showed that, during almond pollination, both hive entrance exposure and hive strength have quantifiable effects on colony foraging behaviour and that these effects combine to regulate the overall foraging activity of the pollinating colonies.

Pollen products collected from honey bee hives experiencing minor stress have altered fungal communities and reduced antimicrobial properties.

Fungi are increasingly recognized to play diverse roles within honey bee hives, acting as pathogens, mutualists, and commensals. Pollen products, essential for hive nutrition, host significant fungal communities with potential protective and nutritional benefits. In this study, we profile the fungal communities and antifungal properties of three pollen products from healthy and stressed hives: fresh pollen collected by forager bees from local plants; stored pollen packed into the comb inside the hive; and bee bread, which is stored pollen following anaerobic fermentation used for bee and larval nutrition. Using amplicon sequencing, we found significant differences in fungal community composition, with hive health and sample type accounting for 8.8% and 19.3% of variation in beta diversity, respectively. Pollen and bee bread extracts had species-specific antimicrobial activity and inhibited the fungal hive pathogens Ascosphaera apis, Aspergillus flavus, and Aspergillus fumigatus, and the bacterial hive pathogen Paenibacillus larvae. Activity was positively correlated with phenolic and antioxidant content and was diminished in stressed hives. The plant source of pollen determined by amplicon sequencing differed in stressed hives, suggesting altered foraging behaviour. These findings illustrate the complex interplay between honey bees, fungal communities, and hive products, which should be considered in hive management and conservation.

Treatment of patients with evidence of mast cell-mediated itch in the absence of hives with omalizumab.

PURPOSE: The presence of wheals or hives has been viewed as a hallmark symptom of urticaria, a highly debilitating disease. This study explores our experience with omalizumab in patients with apparent mast-cell mediated pruritus in the absence of hives. MATERIALS AND METHODS: This is a retrospective case series examining all patients with mast cell-mediated pruritus in the absence of hives from April 2022 to May 2024 at a tertiary referral clinic at Icahn School of Medicine at Mount Sinai in New York. Peak pruritus-numerical rating scale (PP-NRS) itch score changes over time were recorded and analyzed. RESULTS: Six patients (67% women; mean [SD] age, 47.67 [13.52] years) were included in the analysis. The median [IQR] pruritus PP-NRS itch score before omalizumab injection was 9 [6 - 10] and the final median [IQR] PP-NRS itch score was 2.5 [0 - 5]. The mean [SD] reduction in the PP-NRS itch score was 6 [3.16]. CONCLUSIONS: This study suggests that patients with evidence of mast cell-mediated pruritus can be identified based on clinical features and may benefit from omalizumab therapy.

Melipona stingless bees and honey microbiota reveal the diversity, composition, and modes of symbionts transmission.

The Melipona gut microbiota differs from other social bees, being characterized by the absence of crucial corbiculate core gut symbionts and a high occurrence of environmental strains. We studied the microbial diversity and composition of three Melipona species and their honey to understand which strains are obtained by horizontal transmission (HT) from the pollination environment, represent symbionts with HT from the hive/food stores or social transmission (ST) between nestmates. Bees harbored higher microbial alpha diversity and a different and more species-specific bacterial composition than honey. The fungal communities of bee and honey samples are also different but less dissimilar. As expected, the eusocial corbiculate core symbionts Snodgrassella and Gilliamella were absent in bees that had a prevalence of Lactobacillaceae - including Lactobacillus (formerly known as Firm-5), Bifidobacteriaceae, Acetobacteraceae, and Streptococcaceae - mainly strains close to Floricoccus, a putative novel symbiont acquired from flowers. They might have co-evolved with these bees via ST, and along with environmental Lactobacillaceae and Pectinatus (Veillonellaceae) strains obtained by HT, and Metschnikowia and Saccharomycetales yeasts acquired by HT from honey or the pollination environment, including plants/flowers, possibly compose the Melipona core microbiota. This work contributes to the understanding of Melipona symbionts and their modes of transmission.

Enhancing monitoring of suspicious activities with AI-based and big data fusion.

This study provides an AI-based detection tool for the surveillance of suspicious activities using data fusion. The system leverages time, location, and specific data pertaining to individuals, objects, and vehicles associated with the agency. The study's training data was obtained from Thailand's military institution. The study focuses on comparing the efficiency between MySQL and Apache Hive for big data processing. According to the findings, MySQL is better suited for quick data retrieval and low storage capacity, while Hive demonstrates higher scalabilities for larger datasets. Furthermore, the study explores the practical utilization of web applications interfaces, enabling real-time display, analysis, and identification suspicious activity results. The web application, built with NuxtJS and MySQL, includes statistics charts and maps that show the status of suspicious items, cars, and people, as well as data filtering options. The system utilizes machine-learning algorithms to train the suspicious identification model, with the best-performing algorithms being the decision tree, reaching 98.867% classification accuracy.

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.

Comparison of surface adsorption efficacies of eco-sustainable agro/animal biomass-derived activated carbon for the removal of rhodamine B and hexavalent chromium.

Effective management and remediation strategies are crucial to minimize the impacts of both organic and inorganic contaminants on environmental quality and human health. This study investigates a novel approach utilizing cotton shell activated carbon (CSAC), rice husk activated carbon (RHAC), and wasp hive activated carbon (WHAC), produced through alkali treatment and carbonization under N2 atmosphere at 600 °C. The adsorption capacities of biomass-derived mesoporous activated carbons (CSAC, RHAC, WHAC) alongside macroporous commercial activated carbons (CAC) were evaluated for removing rhodamine B (Rh B) and hexavalent chromium (Cr6+). The CSAC exhibits remarkable adsorption efficiency (255.4 mg.g-1) for Cr(VI) removal, while RHAC demonstrates superior efficacy (174.2 mg.g-1) for Rh B adsorption. Investigating various optimal parameters including initial pH (pH 3 for Cr and pH 7 for Rh B), catalyst dosage (200 mg.L-1), and initial concentration (20 mg.L-1), the Redlich-Peterson isotherm model is applied to reveal a hybrid adsorption mechanism encompassing monolayer (chemisorption) and multilayer (van der Waals adsorption) processes. Kinetic analysis highlights the pseudo-second-order and Elovich models as the most suitable, suggesting physiochemisorption mechanisms. Thermodynamic analysis indicates the endothermic nature of the adsorption process, with increased randomness at the solid-solution interface. Isosteric heat investigations using Clausius-Clapeyron, Arrhenius, and Eyring equations reveal a heterogeneous surface nature across all activated carbons. Further confirmation of Rh B and Cr(VI) adsorption onto activated carbons is provided through FTIR, FESEM, and EDAX analysis. This study highlights the innovation and promise of utilizing biomass-derived activated carbons for effective pollutant removal.

Winter Hive Debris Analysis Is Significant for Assessing the Health Status of Honeybee Colonies (Apis mellifera).

Honeybee diseases are one of the most significant and most common causes of honeybee colonies' weakness and death. An early diagnosis of subclinical infections is necessary to implement precautionary and control measures. Sampling debris from hive bottom boards is simple, non-invasive, and cheap. In this study, we collected winter debris samples in apiaries located in the continental part of Croatia. We used molecular methods, PCR and qPCR, for the first time to analyze those samples. Laboratory results were compared with the health condition and strength of honeybee colonies at an apiary in spring. Our study successfully identified the presence and quantity of various pathogens, including the presence of Vairimorpha spp. (Nosema spp.), quintefied Paenibacillus larvae, Acute Bee Paralysis Virus (ABPV), Black Queen Cell Virus (BQCV), Deformed Wing Virus (DWV), and Sacbrood Virus (SBV). However, our analysis did not detect Melissococcus plutonius, Crithidia mellificae, Lotmaria passim, and Aethina tumida. Samples of winter debris were also examined for the presence and quantification of the V. destructor mites, and their natural mite fall was observed in spring. Honeybee colonies were simultaneously infected by an average of four to six pathogens. Some observed honeybee colonies developed characteristic symptoms, while others did not survive the winter.

Comparison of Single Cell Transcriptome Sequencing Methods: Of Mice and Men.

Single cell RNAseq has been a big leap in many areas of biology. Rather than investigating gene expression on a whole organism level, this technology enables scientists to get a detailed look at rare single cells or within their cell population of interest. The field is growing, and many new methods appear each year. We compared methods utilized in our core facility: Smart-seq3, PlexWell, FLASH-seq, VASA-seq, SORT-seq, 10X, Evercode, and HIVE. We characterized the equipment requirements for each method. We evaluated the performances of these methods based on detected features, transcriptome diversity, mitochondrial RNA abundance and multiplets, among others and benchmarked them against bulk RNA sequencing. Here, we show that bulk transcriptome detects more unique transcripts than any single cell method. While most methods are comparable in many regards, FLASH-seq and VASA-seq yielded the best metrics, e.g., in number of features. If no equipment for automation is available or many cells are desired, then HIVE or 10X yield good results. In general, more recently developed methods perform better. This also leads to the conclusion that older methods should be phased out, and that the development of single cell RNAseq methods is still progressing considerably.

Uncovering the spatial pattern of invasion of the honeybee pest small hive beetle, Aethina tumida, in Italy

ABSTRACT The fast tracking of invasion spatial patterns of alien species is crucial for the implementation of preventive and management strategies of those species. Recently, a honeybee pest, the small hive beetle Aethina tumida (hereafter SHB), has been reported in Italy, where it colonized more than 50 apiaries in an area of about 300 km2. SHB is a nest parasite and scavenger of honeybee colonies native of Sub-Saharian Africa. Likely being helped by the globalization of apiculture, SHB underwent several invasions in the last twenty years, causing locally relevant economic impact. While many features of its biology have been addressed, an important knowledge gap concerns the spatial invasion dynamics in invaded areas. In this paper we coupled two spatial analysis techniques (geographic profiling and a density-based spatial clustering algorithm) to uncover the possible invasion pattern of SHB in Italy. We identified the port town of Gioia Tauro as the most likely point from which SHB may have spread and suggested the possible successive axes of diffusion. These putative diffusion paths suggest that the SHB spread in south Italy might have been due to a mix of natural dispersal between close apiaries and longer distance movement through faster, likely human-mediated, communication routes.

Additional information on reproductive behavior of the Red-breasted Toucan, Ramphastos dicolorus (Aves: Piciformes: Ramphastidae) / Informações adicionais sobre o comportamento reprodutivo do tucano-de-bico-verde, Ramphastos dicolorus (Aves: Piciformes: Ramphastidae)

Abstract: Ramphastidae is Family of secondary cavity-nesting birds that is endemic to the Neotropical region. Here we provide a comprehensive literature review of nest measurements, and new information on characteristics of the nest cavity, parental nest attendance, incubation and nestling period of the Red-breasted Toucan, Ramphastos dicolorus. We also add information on the origin of the seeds found inside toucan nests and propose a potential function for them. Eight nests were studied in three cavities during six breeding seasons at Parque Estadual das Fontes do Ipiranga, São Paulo, and at a private allotment in Ibiúna, southeastern Brazil. The reproductive activities of the birds were observed from September to February. In one cavity, we found the first record of a Meliponini bee hive (Scaptotrigona bipunctata) co-existing with a toucan nest. At the beginning of the nesting season, the parents regurgitate seeds on the floor of the cavity, covering it with them. This preparation plays a role in nest sanitation, once the seeds act as a substrate for the feces facilitating the process of carrying them off the nest by the female. The incubation and nestling periods lasted 17.2 ± 1.2 and 43 ± 5.4 days, respectively, and both parents shared egg incubation, nestling, brooding and provisioning, but only the female performed nest sanitation. This differs from published observations on other species of the group, suggesting that the participation of each sex in nest attendance may not be the same for all ramphastids. Additionally, our observations suggest that selecting anthropic locations for nesting can negatively influence toucan nest success.

Resumo: Ramphastidae compreende uma Família de aves que nidificam em cavidades, são endêmicas da região Neotropical, mas possuem os aspectos relacionados à sua biologia reprodutiva ainda pouco conhecidos. No presente estudo, nós fornecemos uma revisão detalhada da literatura acerca de medidas de ninhos, características das cavidades, cuidado parental e períodos de incubação e permanência do tucano-de-bico-verde Ramphastos dicolorus. Nós também descrevemos e propomos uma possível função para as sementes encontradas dentro dos ninhos dessa espécie. Foram estudados oito ninhos, em três diferentes cavidades durante seis temporadas reprodutivas no Parque Estadual das Fontes do Ipiranga, São Paulo, e em um loteamento particular em Ibiúna, ambos no sudeste do Brasil. As atividades reprodutivas foram observadas de Setembro a Fevereiro. Em uma das cavidades foi registrada a coexistência entre uma colmeia de abelhas Meliponini Scaptotrigona bipunctata e um ninho de tucanos, sendo este o primeiro registro desse tipo de interação para Ramphastidae. No início do período de nidificação os parentais regurgitavam sementes no fundo da cavidade, cobrindo-o, e nossas observações mostram que essa preparação desempenha o papel de facilitar na limpeza do ninho, uma vez que as sementes servem como substrato para as fezes e possibilitam que a fêmea possa carrega-las para fora. Os períodos de incubação e permanência dos filhotes no ninho duraram 17,2 ± 1,2 e 43 ± 5,4 dias, respectivamente, e ambos os parentais dividiram a incubação dos ovos, aquecimento e alimentação dos ninhegos, mas somente a fêmea realizou a limpeza do ninho. Entretanto, tais comportamentos observados para o tucano-de-bico-verde são diferentes do que se conhece para outras espécies do grupo, sugerindo que o investimento de cada membro do casal no atendimento ao ninho pode não ser o mesmo para todos os ramphastídeos. As observações realizadas também mostram que a seleção de locais antropizados para nidificar poderia influenciar negativamente o sucesso reprodutivo dos tucanos.