New Medical Herbalism Formulations as a Targeted Therapeutic Strategies Used in Southern Tunisia to Promote Neo-Epithelium, Angiogenesis, and Collagen Biosynthesis and to Impede Scar Formation Post-Third-Degree Burned Skin.

The aim of the study is to assess the suitability of the herbal formulation for topical application as a skin burn dressing on the in vivo wound-closure of third-degree wound injuries. Rat wound models were used to prove the in vivo skin burn-healing process. Body weight gain, food and water intake, and behavior were investigated daily during treatment period. Cutaneous biopsies of the burned wound surfaces were monitored at days 4, 13, and 28. Formulation markedly (P < .05) increased wound repair rate and collagen production compared to untreated burnt skin. Macroscopic and histological analysis of the wound of formula (F)-treated group showed significant skin contraction rate and rapid wound healing without scar through regeneration of epidermis that were approved in formula mixed with honey (F-hY)- and Drs-treated wound compared with thymol, and the untreated wound tissues that were not covered by denuded epithelial. Furthermore, the wound healing efficacy of F-hY, F, and Drs cream was proved by decreased the amount of malondialdehyde compared to untreated rats. In conclusion, F and F-hY was found to promote cutaneous wound repair. In all case, the formula alone or mixed with honeybees was even better than thymol in the repair of cutaneous wound.

A Modeling Approach for Assessing Ecological Risks of Neonicotinoid Insecticides from Emission to Nontarget Organisms: A Case Study of Cotton Plant.

The use of neonicotinoid insecticides in agriculture has posed threats to ecological systems, and there is a need to assess the ecological risks of neonicotinoids from emission to nontarget organisms. We introduced a modeling approach to assess the ecological risks of neonicotinoids using honeybee and earthworm as model organisms, and the simulation was flexible under different environmental conditions. Using the cotton plant as an example, the simulation results demonstrated that under current recommended application rates, the use of common neonicotinoid insecticides posed no threat to earthworms, with the simulated risk quotients (RQs) much lower than 1. However, the simulation for some neonicotinoid insecticides (e.g., acetamiprid) indicated that using these insecticides on cotton plants could threaten honeybees, with simulated RQs higher than 1. The variability analysis showed that in high-latitude regions, the unacceptable risk to honeybees posed by insecticide application can be further elevated due to cold, wet weather that results in relatively high insecticide levels in pollen and nectar. The model evaluation showed large overlaps of simulated risk intervals between the proposed and existing (BeeREX) models. Because the proposed and existing models have different simulation mechanisms, we recommend that these two models be used together to complement each other in future studies. Environ Toxicol Chem 2023;42:928-938. © 2023 SETAC.

Managed honeybees decrease pollination limitation in self-compatible but not in self-incompatible crops.

Modern agriculture is becoming increasingly pollinator-dependent. However, the global stock of domesticated honeybees is growing at a slower rate than its demand, while wild bees are declining worldwide. This uneven scenario of high pollinator demand and low pollinator availability can translate into increasing pollination limitation, reducing the yield of pollinator-dependent crops. However, overall assessments of crop pollination limitation and the factors determining its magnitude are missing. Based on 52 published studies including 30 crops, we conducted a meta-analysis comparing crop yield in pollen-supplemented versus open-pollinated flowers. We assessed the overall magnitude of pollination limitation and whether this magnitude was influenced by (i) the presence/absence of managed honeybees, (ii) crop compatibility system (i.e. self-compatible/self-incompatible) and (iii) the interaction between these two factors. Overall, pollen supplementation increased yield by approximately 34%, indicating sizable pollination limitation. Deployment of managed honeybees and self-compatibility were associated with lower pollination limitation. Particularly, active honeybee management decreased pollination limitation among self-compatible but apparently not among self-incompatible crops. These findings indicate that current pollination regimes are, in general, inadequate to maximize crop yield, even when including managed honeybees, and stress the need of transforming the pollination management paradigm of agricultural landscapes.

Sweeping-micellar electrokinetic chromatography with tandem mass spectrometry as an alternative methodology to determine neonicotinoid and boscalid residues in pollen and honeybee samples.

In this work, it is proposed for the first time an electrophoretic approach based on micellar electrokinetic chromatography coupled with tandem mass spectrometry (MEKC-MS/MS) for the simultaneous determination of nine neonicotinoids (NNIs) together with the fungicide boscalid in pollen and honeybee samples. The separation was performed using ammonium perfluorooctanoate (50 mM, pH 9) as both volatile surfactant and electrophoretic buffer compatible with MS detection. A stacking strategy for accomplishing the on-line pre-concentration of the target compounds, known as sweeping, was carried out in order to improve separation efficiency and sensitivity. Furthermore, a scaled-down QuEChERS was developed as sample treatment, involving a lower organic solvent consumption and using Z-Sep+ as dispersive sorbent in the clean-up step. Regarding the detection mode, a triple quadrupole mass spectrometer was operating in positive ion electrospray mode (ESI+) under multiple reaction monitoring (MRM). The main parameters affecting MS/MS detection as well as the composition of the sheath-liquid (ethanol/ultrapure water/formic acid, 50:49.5:0.5 v/v/v) and other electrospray variables were optimized in order to achieve satisfactory sensitivity and repeatability. Procedural calibration curves were established in pollen and honeybee samples with LOQs below 11.6 µg kg-1 and 12.5 µg kg-1, respectively. Precision, expressed as RSD, lower than 15.2% and recoveries higher than 70% were obtained in both samples. Two positive samples of pollen were found, containing imidacloprid and thiamethoxam. Imidacloprid was also found in a sample of honeybees. The obtained results highlight the applicability of the proposed method, being an environmentally friendly, efficient, sensitive and useful alternative for the determination of NNIs and boscalid in pollen and honeybee samples.

Beehive products as bioindicators of antimicrobial resistance contamination in the environment.

The use of antimicrobials in agricultural, veterinary and medical practice exerts selective pressure on environmental microbiota, promoting the emergence and spread of antimicrobial resistance (AMR), a global concern for the One Health Initiative Task Force (OHITF). Honeybees have been studied as bioindicators of AMR in the environment, but little is known about beehive products like honey and pollen. The aim of this study was to assess the prevalence of AMR genes (ARGs) in beehive products and investigated their origins. Specifically, possible associations between ARGs, microbiota and other characteristics of different honey and pollen samples, including country of origin, flower type, type of commercial distribution and environmental factors, such as land use, weather and composition of the environment surrounding the beehives were investigated. We found that beehive products harboured ARGs conferring resistance to ß-lactams, macrolides, (fluoro)quinolones and polymyxins. Most samples possessed resistance to multiple antimicrobial classes, with honey and pollen showing similar ARG profiles. Even if Lactobacillus and Acinetobacter genera were common in the microbial communities of both honey and pollen, Bacillus, Clostridium, and Bombella defined honey microbiota, while Pseudomonas and Vibrio were enriched in pollen. ErmB and blaTEM-1 co-occurred with Lactobacillus and Fructobacillus, while positive associations between ß-lactams and macrolides and anthropogenic environments (i.e. industrial and commercial areas and non-irrigated arable lands) were found. Altogether, our findings suggest that ARGs in honey and pollen might originate from the honeybee foraging environment, and that the beehive products can be used as bioindicators of the AMR environmental contamination.

Thin-layer chromatography in the authenticity testing of bee-products.

Quality control, nutritional value and the monitoring of hazardous residues in honey bee- products have become major topics for both producers and consumers. Due to its potential role in human health, bee-products rich in bioactive compounds are becoming increasingly popular. This review aims to provide an overview of thin-layer chromatography methods used in quality control,authenticity testing and chemical profiling of bee-products in order to help scientists engaged in the field of bee-products chemistry to utilize the advantages of this technique in the detection and elimination of fraudulent practices in bee-product manufacturing. Recently, hyphenation of thin-layer chromatography, image analysis and chemometrics support bee-products analysisbysimultaneousdeterminationofanalytes with different detection principles, identification of individual bioactive compounds as well as structure elucidation of compounds. Highlighted opportunities of thin-layer chromatography could encourage further investigations that would lead to improvements in the detection and elimination of marketing fraudulent practices.

Varroa destructor from the Laboratory to the Field: Control, Biocontrol and IPM Perspectives-A Review.

Varroa destructor is a real challenger for beekeepers and scientists: fragile out of the hive, tenacious inside a bee colony. From all the research done on the topic, we have learned that a better understanding of this organism in its relationship with the bee but also for itself is necessary. Its biology relies mostly on semiochemicals for reproduction, nutrition, or orientation. Many treatments have been developed over the years based on hard or soft acaricides or even on biocontrol techniques. To date, no real sustainable solution exists to reduce the pressure of the mite without creating resistances or harming honeybees. Consequently, the development of alternative disruptive tools against the parasitic life cycle remains open. It requires the combination of both laboratory and field results through a holistic approach based on health biomarkers. Here, we advocate for a more integrative vision of V. destructor research, where in vitro and field studies are more systematically compared and compiled. Therefore, after a brief state-of-the-art about the mite's life cycle, we discuss what has been done and what can be done from the laboratory to the field against V. destructor through an integrative approach.

Honeybee Exposure to Veterinary Drugs: How Is the Gut Microbiota Affected?

Several studies have outlined that a balanced gut microbiota offers metabolic and protective functions supporting honeybee health and performance. The present work contributes to increasing knowledge on the impact on the honeybee gut microbiota of the three most common veterinary drugs (oxytetracycline, sulfonamides, and tylosin). The study was designed with a semi-field approach in micro-hives containing about 500 honeybees. Micro-hives were located in an incubator during the day and moved outdoors in the late afternoon, considering the restrictions on the use of antibiotics in the open field but allowing a certain freedom to honeybees; 6 replicates were considered for each treatment. The absolute abundance of the major gut microbial taxa in newly eclosed individuals was studied with qPCR and next-generation sequencing. Antimicrobial resistance genes for the target antibiotics were also monitored using a qPCR approach. The results showed that the total amount of gut bacteria was not altered by antibiotic treatment, but qualitative variations were observed. Tylosin treatment determined a significant decrease of α- and ß-diversity indices and a strong depletion of the rectum population (lactobacilli and bifidobacteria) while favoring the ileum microorganisms (Gilliamella, Snodgrassella, and Frischella spp.). Major changes were also observed in honeybees treated with sulfonamides, with a decrease in Bartonella and Frischella core taxa and an increase of Bombilactobacillus spp. and Snodgrassella spp. The present study also shows an important effect of tetracycline that is focused on specific taxa with minor impact on alfa and beta diversity. Monitoring of antibiotic resistance genes confirmed that honeybees represent a great reservoir of tetracycline resistance genes. Tetracycline and sulfonamides resistance genes tended to increase in the gut microbiota population upon antibiotic administration. IMPORTANCE This study investigates the impact of the three most widely used antibiotics in the beekeeping sector (oxytetracycline, tylosin, and sulfonamides) on the honeybee gut microbiota and on the spread of antibiotic resistance genes. The research represents an advance to the present literature, considering that the tylosin and sulfonamides effects on the gut microbiota have never been studied. Another original aspect lies in the experimental approach used, as the study looks at the impact of veterinary drugs and feed supplements 24 days after the beginning of the administration, in order to explore perturbations in newly eclosed honeybees, instead of the same treated honeybee generation. Moreover, the study was not performed with cage tests but in micro-hives, thus achieving conditions closer to real hives. The study reaches the conclusion that the most common veterinary drugs determine changes in some core microbiota members and that incidence of resistance genes for tetracycline and sulfonamides increases following antibiotic treatment.

Assessment of the impacts of microbial plant protection products containing Bacillus thuringiensis on the survival of adults and larvae of the honeybee (Apis mellifera).

This study was aimed at evaluating the effect of a microbial pest-controlling product (MPCP) with the active substance Bacillus thuringiensis ssp. aizawai (strain: ABTS-1857) on adults and larvae of honeybees. To determine the contamination levels of Bt spores in different matrices, a colony-feeding study under semi-field conditions was performed. Furthermore, two chronic adult trials and a chronic larval study were conducted under laboratory conditions to test the effects of different concentrations of the plant protection product (PPP) on the development and mortality. Possible modifications of the chronic oral toxicity test were assessed by additional pollen feeding. Our results showed that Bt spores were detected in all matrices over the entire test duration in different concentrations, decreasing over time. The survival of adult bees and larvae was negatively affected in laboratory conditions after a chronic exposure to the MPCP depending on the tested concentrations. Moreover, the earliest sign of bee mortality, resulting from exposure to ABTS-1857, was recorded only after 96 h at the highest tested concentration. Pollen feeding to adults significantly increased the survival of the treated bees. In conclusion, the PPP with the Bt strain ABTS-1857 showed an effect on the mortality of adults and larvae under laboratory conditions. Further studies with Bt-based PPPs under realistic field conditions are necessary to evaluate the potential risk of those MPCPs on honeybees.

Bee products as a source of promising therapeutic and chemoprophylaxis strategies against COVID-19 (SARS-CoV-2).

The emergence of novel coronavirus (SARS-CoV-2) in 2019 in China marked the third outbreak of a highly pathogenic coronavirus infecting humans. The novel coronavirus disease (COVID-19) spread worldwide, becoming an emergency of major international concern. However, even after a decade of coronavirus research, there are still no licensed vaccines or therapeutic agents to treat the coronavirus infection. In this context, apitherapy presents as a promising source of pharmacological and nutraceutical agents for the treatment and/or prophylaxis of COVID-19. For instance, several honeybee products, such as honey, pollen, propolis, royal jelly, beeswax, and bee venom, have shown potent antiviral activity against pathogens that cause severe respiratory syndromes, including those caused by human coronaviruses. In addition, the benefits of these natural products to the immune system are remarkable, and many of them are involved in the induction of antibody production, maturation of immune cells, and stimulation of the innate and adaptive immune responses. Thus, in the absence of specific antivirals against SARS-CoV-2, apitherapy could offer one hope toward mitigating some of the risks associated with COVID-19.

Concentration- and time-dependent toxicity of commonly encountered pesticides and pesticide mixtures to honeybees (Apis mellifera L.).

Honeybees are exposed to a wide range of pesticides for long periods via contaminated water, pollen and nectar. Some of those pesticides might constitute health hazards in a time- and dose-dependent manner. Time-dependent toxicity profiles for many applied pesticides are lacking, despite the fact that such profiles are crucial for toxicological evaluations. Therefore, we sought to determine the time-dependent toxicities of pesticides/pesticide metabolites frequently found in Israeli beehives, namely, amitraz metabolites, N'-(2,4-dimethylphenyl)-N-methylformamidine (DMPF) and N-(2,4-dimethylphenyl)-formamide (DMF), coumaphos, imidacloprid, thiacloprid, acetamiprid and dimethoate (toxic reference). By applying accepted methodological approaches such as the modified Haber's rule (product of concentration and exposure duration leads to a constant effect) and comparisons between cumulative doses at different time points, we determined the time-dependent toxicities of these pesticides. We also studied the mixture toxicities of frequently occurring pesticide combinations and estimated their potential contributions to the overall toxicities of neonicotinoids. Thiacloprid was the only pesticide that complied with Haber's rule. DMPF, dimethoate and imidacloprid exhibited time-diminished -toxicities. In contrast, DMF and acetamiprid exhibited time-reinforced toxicities. Neither the binary mixtures nor the tertiary mixtures of DMF, DMPF and coumaphos at 10 times their environmentally relevant concentrations potentiated the neonicotinoids' toxicities. DMPF and imidacloprid were found to present the greatest hazard to honeybees, based on their 50% lethal cumulative dose and 50% lethal time. Amitraz's instability, its low detection frequency and high toxicity profile of its metabolite, DMPF, lead us to the conclusion that DMPF constitutes the actual toxic entity responsible for amitraz's toxic effect.

Drying Techniques and Storage: Do They Affect the Nutritional Value of Bee-Collected Pollen?

In this study, the effect of different drying processes (freeze-drying (FD), microwave-assisted drying (MWD) and classic hot air drying (HAD)) on the polyphenols, flavonoids, and amino acids content was investigated on bee-collected chestnut, willow and ivy pollen for human consumption. Furthermore, the pollen chemical properties were monitored after three and six months of storage, and then analyzed using a multivariate approach. Chestnut pollen was the richest source of polyphenols, flavonoids, and rutin, while ivy pollen contained the highest amount of total and free amino acids, and total and free proline. Drying and storage affected pollen chemical composition with species-dependent effects. MWD allowed the best retention of flavonoids in chestnut pollen for up to six months of storage. All drying techniques led to a depletion of flavonoids in willow pollen; however, MWD ensured the highest flavonoids content after six months. FD and MWD did not lead to flavonoids depletion in ivy pollen during storage. Additionally, storage did not affect the rutin content, which was highest in FD willow samples after six months. Notably, both FD and MWD techniques are efficient in preserving amino acids-related quality of bee pollen up to six months of storage.

Bees and the Environmental Impact of the Rupture of the Fundão Dam.

The environmental consequences associated with the Fundão tailings dam failure in Mariana, Minas Gerais, Brazil on 5 November 2015 are still being investigated. Bees are the main pollinators believed to be most affected by the accident because they occur throughout the area affected by the dam rupture and build their nests in the soil. In this study, we evaluated the richness of different bee species in areas affected and not affected by the accident and measured the concentration of different metals on the pollen basket, or corbicula, located on the hind legs of Apis mellifera. Diversity indices were similar and there was no statistical difference in the diversity of bees sampled when comparing the sites affected and not affected by the tailings mud, either before or after the dam rupture. It is possible that the similarity is due to nearby forests that may be serving as areas of refuge. The levels of Al, Cu, Cd, Hg, and Pb in A. mellifera suggest no change in the level of metals in pollen collected by the bees that can be attributed to the rupture of the dam. Integr Environ Assess Manag 2020;16:631-635. © 2020 SETAC.

As consequências ambientais da ruptura da barragem de Fundão, ocorrida em Mariana, Minas Gerais, Brasil, em 5 de novembro de 2015, ainda estão sendo investigadas. As abelhas, principal grupo de polinizadores, podem ter sido afetadas direta e indiretamente, uma vez que a maioria das espécies constrói seus ninhos no solo. Neste estudo, avaliamos a riqueza de espécies de abelhas nas áreas afetadas e não afetadas e medimos a concentração de metais pesados nas corbículas de Apis mellifera, espécie que ocorre em toda a área afetada pela ruptura da barragem. Os índices de diversidade foram semelhantes e não houve diferença estatística na diversidade de abelhas amostradas quando comparamos locais afetados e não afetados pela lama de rejeitos após o rompimento da barragem. É possível que a semelhança se deva a florestas próximas que possam estar servindo como áreas de refúgio. Os resultados mostraram a presença de Al, Cu, Cd, Hg e Pb em A. mellifera. Nossos resultados indicam que não há efeito da contaminação de metais pesados nas abelhas devido à ruptura da barragem. Integr Environ Assess Manag 2020;16:631-635.

Microscopic investigation of palyno-morphological features of melliferous flora of Lakki Marwat district, Khyber Pakhtunkhwa, Pakistan.

The present study was carried out to investigate the polliniferous bee flora of Lakki Marwat district, Khyber Pakhtunkhwa, Pakistan. A total of 18 species, belonging to 13 different families were identified for the foraging activities of honeybees, namely, Cenchrus biflorus, Cestrum nocturnum, Citrus limon, Combretum indicum, Datura innoxia, Duranta erecta, Hamelia patens, Helianthus annuus, Ipomoea cairica, Luffa aegyptiaca, Nerium oleander, Ocimum basilicum, Parthenium hysterophorus, Pennisetum typhoides, Prosopis cineraria, Prosopis juliflora, Saccharum spontaneum, and Ziziphus jujuba. The identified melliferous flora was collected for the morpho-palynological investigation. Pollen were acetolyzed, measured, and described qualitatively using light microscopy. The pollen types varied from psilate to echinate and colpi from tricolpate to hexacolpate. Quantitative characteristics including polar diameter, equatorial diameter, polar to equatorial ratio, colpi length, colpi width, spine length, spine width, and exine thickness of the pollen were determined and analyzed statistically using software IBM SPSS Statistics 20. The results of the present study highlighted the significance of pollen morphology of the bee flora of the area which may lead to the identification of the potential and useful botanical sources for beekeepers and to check the honey quality marketed from the study area.

Microbiota of Honeybees, Apis mellifera Adansonii (Hymenoptera: Apidae) from Selected Ecozones, South West Nigeria.

BACKGROUND AND OBJECTIVES: Honeybees explore their environment in search of nectar and pollens including water whose sources are contaminated by variety of microbes. This has both negative and positive economic implications on health status of honeybees, the consumers of the products including the quality of the products. Hence, the study was designed to investigate the effect of geographical locations on the occurrence and diversity of microbiota on the external integument and the digestive gut of adult worker honeybees, Apis mellifera adansonii. MATERIALS AND METHODS: The honeybee samples were sourced from Ondo, Ogun, Oyo and Osun of South West Nigeria. The samples were subjected to microbiological analyses using standard techniques. Two-ways ANOVA was used for statistical test of the data. RESULTS: The microbiota of the studied samples includes Corynebacterium kutsceri, Corynebacterium xerosis, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus fermentum, Micrococcus luteus, Micrococcus varians and Staphylococcus aureus which were gram-positive bacteria, while gram-negative bacteria include Aeromonas veronii, Citrobacter diversus, Citrobacter freundii, Klebsiella oxytoca, Klebsiella pneumonia and Pseudomonas aeruginosa. Fungi isolates include Aspergillus parasiticus, Aspergilus flavus, Aspergilus fumigatus, Aspergilus niger and Rhizopus stolonifer. Most microbiota were not cosmopolitan across selected ecozones of south west, Nigeria except for Aspergillus fumigatus. The number of identified species of microbiota either in the digestive guts or on the external integument of worker honeybees from selected ecozones varies from one ecozone to another. There was significant species diversity of microbiota on external integument than in digestive guts of adult workers of A. mellifera adansonii from selected ecozones in South West, Nigeria. CONCLUSION: The study showed that both the external integument and the digestive guts of A. mellifera adansonii exhibited variety of microbial flora providing research exploration opportunity for probable microbes of economic and geographical importance.

Detection of Pesticide Residues in Selected Bee Products of Honeybees (Apis melllifera L.) Colonies in a Preliminary Study from Seychelles Archipelago.

Pesticide residues in honey and pollen from Seychelles against a target of 108 pesticides using LC-MS/MS were analyzed. Fifteen pesticides were detected, at trace levels (< 15 ppb) and below the acceptable maximum residue limits (MRLs) as per EU regulations. In honey, six insecticide and three fungicide residues were detected. Eight insecticide and four fungicide residues were detected in the pollen matrix. The least contaminated honey and pollen samples had three and nine chemical residues respectively while the most contaminated honey and pollen samples had eight and eleven chemical residues respectively. Contact and oral LD50 values were used to calculate Pollen Hazard Quotients (PHQ) = concentration in ppb ÷ LD50 as µg/bee. The pollen hazard quotients (PHQ) obtained are way below those reported in literature. Residues were detected in low quantities, however, their high frequency and diversity and possible synergistic interactions may lead to negative impact on honeybees' health in Seychelles.

Royal Jelly: An ancient remedy with remarkable antibacterial properties.

Royal Jelly (RJ), a honeybee hypopharyngeal gland secretion of young nurse and an exclusive nourishment for bee queen, has been used since ancient times for care and human health and it is still very important in traditional and folkloristic medicine, especially in Asia within the apitherapy. Recently, RJ and its protein and lipid components have been subjected to several investigations on their antimicrobial activity due to extensive traditional uses and for a future application in medicine. Antimicrobial activities of crude Royal Jelly, Royalisin, 10-hydroxy-2-decenoic acid, Jelleines, Major Royal Jelly Proteins against different bacteria have been reported. All these beehive products showed antimicrobial activities that lead their potential employment in several fields as natural additives. RJ and its derived compounds show a highest activity especially against Gram positive bacteria. The purpose of this Review is to summarize the results of antimicrobial studies of Royal Jelly following the timescale of the researches. From the first scientific applications to the isolation of the single components in order to better understand its application in the past years and propose an employment in future studies as a natural antimicrobial agent.

How regulation based on a common stomach leads to economic optimization of honeybee foraging.

Simple regulatory mechanisms based on the idea of the saturable 'common stomach' can control the regulation of protein foraging and protein allocation in honeybee colonies and colony-level responses to environmental changes. To study the economic benefits of pollen and nectar foraging strategies of honeybees to both plants and honeybees under different environmental conditions, a model was developed and analyzed. Reallocation of the foraging workforce according to the quality and availability of resources (an 'adaptive' strategy used by honeybees) is not only a successful strategy for the bees but also for plants, because intensified pollen foraging after rain periods (when nectar quality is low) compensates a major fraction of the pollination flights lost during the rain. The 'adaptive' strategy performed better than the'fixed' (steady, minimalistic, and non-adaptive foraging without feedback) or the 'proactive' (stockpiling in anticipation of rain) strategies in brood survival and or in nectar/sugar economics. The time pattern of rain periods has profound effect on the supply-and-demand of proteins. A tropical rain pattern leads to a shortage of the influx of pollen and nectar, but it has a less profound impact on brood mortality than a typical continental rainfall pattern. Allocating more bees for pollen foraging has a detrimental effect on the nectar stores, therefore while saving larvae from starvation the 'proactive' strategy could fail to collect enough nectar for surviving winter.

Lactic acid bacterial symbionts in honeybees - an unknown key to honey's antimicrobial and therapeutic activities.

Could honeybees' most valuable contribution to mankind besides pollination services be alternative tools against infections? Today, due to the emerging antibiotic-resistant pathogens, we are facing a new era of searching for alternative tools against infections. Natural products such as honey have been applied against human's infections for millennia without sufficient scientific evidence. A unique lactic acid bacterial (LAB) microbiota was discovered by us, which is in symbiosis with honeybees and present in large amounts in fresh honey across the world. This work investigates if the LAB symbionts are the source to the unknown factors contributing to honey's properties. Hence, we tested the LAB against severe wound pathogens such as methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa and vancomycin-resistant Enterococcus (VRE) among others. We demonstrate a strong antimicrobial activity from each symbiont and a synergistic effect, which counteracted all the tested pathogens. The mechanisms of action are partly shown by elucidating the production of active compounds such as proteins, fatty acids, anaesthetics, organic acids, volatiles and hydrogen peroxide. We show that the symbionts produce a myriad of active compounds that remain in variable amounts in mature honey. Further studies are now required to investigate if these symbionts have a potential in clinical applications as alternative tools against topical human and animal infections.

Novel biopesticide based on a spider venom peptide shows no adverse effects on honeybees.

Evidence is accumulating that commonly used pesticides are linked to decline of pollinator populations; adverse effects of three neonicotinoids on bees have led to bans on their use across the European Union. Developing insecticides that pose negligible risks to beneficial organisms such as honeybees is desirable and timely. One strategy is to use recombinant fusion proteins containing neuroactive peptides/proteins linked to a 'carrier' protein that confers oral toxicity. Hv1a/GNA (Galanthus nivalis agglutinin), containing an insect-specific spider venom calcium channel blocker (ω-hexatoxin-Hv1a) linked to snowdrop lectin (GNA) as a 'carrier', is an effective oral biopesticide towards various insect pests. Effects of Hv1a/GNA towards a non-target species, Apis mellifera, were assessed through a thorough early-tier risk assessment. Following feeding, honeybees internalized Hv1a/GNA, which reached the brain within 1 h after exposure. However, survival was only slightly affected by ingestion (LD50>100 µg bee(-1)) or injection of fusion protein. Bees fed acute (100 µg bee(-1)) or chronic (0.35 mg ml(-1)) doses of Hv1a/GNA and trained in an olfactory learning task had similar rates of learning and memory to no-pesticide controls. Larvae were unaffected, being able to degrade Hv1a/GNA. These tests suggest that Hv1a/GNA is unlikely to cause detrimental effects on honeybees, indicating that atracotoxins targeting calcium channels are potential alternatives to conventional pesticides.