This study aims to assess the pollution of honey by heavy metals based on the efforts of an urbanization gradient in the city of Annaba (Northeast Algeria) where the dosage of five heavy metals (Fe, Cr, Ni, Cu, Cd) was carried out in four sites.The level of heavy metals was determined by atomic absorption spectrophotometer. The results obtained indicated that even when all the samples were contaminated they were of good quality since the concentrations did not exceed the international standards.According to the results of heavy metal concentrations in the honeys studied, the most abundant element is Fe with an average concentration of 6.956 ± 2.045 (mg/kg), Cr 0.765 ± 0.197 (mg/kg), Ni 0.6005 ± 0.159 (mg/kg), Cu 0.21025 ± 0.065 (mg/kg) and Cd 0.01425 ± 0.005 (mg/kg).The heavy metals studied are present in all samples but trace amounts. Moreover, the comparison of the honey from the four sites indicates to us that the healthiest honey is that of the urban site.

Chlorinated paraffins (CPs) are industrial chemicals produced in large quantities. Short-chain CPs (SCCPs) were classified as persistent organic pollutants under the Stockholm Convention in 2017. Medium-chain CPs (MCCPs) became candidate persistent organic pollutants in 2021. CPs are now ubiquitously found in the environment. Honey bees can be exposed to CPs during foraging, and this exposure subsequently results in the contamination of honey and other bee products along with colony food production and storage. Here, SCCP and MCCP concentrations in honey collected from Chinese apiaries in 2015 and 2021 were determined. Total CP concentrations in honey from 2021 to 2015 were comparable, but the ratio of MCCPs/SCCPs was higher in 2021 than in 2015. SCCP and MCCP congener group profiles in all honey samples were similar and dominated by C₁₀–₁₁Cl₆–₇ and C₁₄Cl₆–₇, respectively. MCCP concentrations were also higher than SCCP concentrations in bees, pollen, and wax but not in bee bread, which were all collected in 2021. The order of average CP concentrations was determined as wax > bee > pollen > bee bread > honey. Poor relationships were found between SCCP concentrations in honey and other samples, but a relationship between MCCP concentrations in honey and other samples was observed. Migration tests of CPs in plastic bottles showed essentially no migration into honey during storage. The risks to humans from CPs in honey are low.

Pollinating bees are stressed by highly variable environmental conditions, malnutrition, parasites and pathogens, but may also by getting in contact with microorganisms or entomopathogenic nematodes that are used to control plant pests and diseases. While foraging for water, food, or nest material social as well as solitary bees have direct contact or even consume the plant protection product with its active substance (e.g., viruses, bacteria, fungi, etc.). Here, we summarize the results of cage, microcolony, observation hive assays, semi-field and field studies using full-size queen-right colonies. By now, some species and subspecies of the Western and Eastern honey bee (Apis mellifera, A. cerana), few species of bumble bees, very few stingless bee species and only a single species of leafcutter bees have been studied as non-target host organisms. Survival and reproduction are the major criteria that have been evaluated. Especially sublethal effects on the bees' physiology, immune response and metabolisms will be targets of future investigations. By studying infectivity and pathogenic mechanisms, individual strains of the microorganism and impact on different bee species are future challenges, especially under field conditions. Overall, it became evident that honey bees, bumble bees and few stingless bee species may not be suitable surrogate species to make general conclusions for biological mechanisms of bee-microorganism interactions of other social bee species. Solitary bees have been studied on leafcutter bees (Megachile rotundata) only, which shows that this huge group of bees (∼20,000 species worldwide) is right at the beginning to get an insight into the interaction of wild pollinators and microbial plant protection organisms.

Neonicotinoids and the closely related insecticide classes sulfoximines and butenolides have recently attracted growing concerns regarding their potential negative effects on non-target organisms, including pollinators such as bees. Indeed, it is becoming increasingly clear that these effects may occur at much lower levels than those considered to be safe for humans. To properly assess the ecological and environmental risks posed by neonicotinoids, appropriate sampling and analytical procedures are needed. Here, we used honey as reliable environmental sampler and developed an unprecedentedly sensitive method based on QuEChERS and UHPLC-MS/MS for the simultaneous determination of the nine neonicotinoids and related molecules currently present on the market (acetamiprid, clothianidin, dinotefuran, flupyradifurone, imidacloprid, nitenpyram, sulfoxaflor, thiacloprid and thiamethoxam). The method was validated and provided excellent levels of precision and accuracy over a wide concentration range of 3–4 orders of magnitude. Lowest limits of quantification (LLOQs) as low as 2–20 pg/g of honey depending on the analytes were reached. The method was then applied to the analysis of 36 honey samples from various regions of the World which had already been analysed for the five most common neonicotinoids (acetamiprid, clothianidin, imidacloprid, thiacloprid and thiamethoxam) in a previous study. This allowed us to determine the long-term stability (i.e. up to 40 months) of these molecules in honey, both at room temperature and −20 °C. We found that the five pesticides were stable over a period of several years at −20 °C, but that acetamiprid and thiacloprid partially degraded at room temperature. Finally, we also measured the levels of dinotefuran, nitenpyram, sulfoxaflor and flupyradifurone and found that 28% of the samples were contaminated by at least one of these pesticides.

Aim of this study was to determine by Ion Chromatography ions (Na+, Ca++, Mg++, NH4+, Cl−, Br−, SO42−, NO3−, PO43−) in honeys (honeydew and floral nectar honeys) from different Italian Regions and from countries of the Western Balkan area. The compositional data were processed by multivariate analysis (PCA and HCA). Arboreal honeydew honeys from the Western Balkans had higher concentrations (from two to three times) of some environmental pollutants (Br−, SO42− and PO43− contents), due to industrial and agricultural activities, than those from Italian regions. The cationic profiles were very similar in both groups. Multivariate analysis indicated a clear difference between nectar honeys and arboreal/honeydew honeys (recognition of the botanical origin). These findings point to the potential of ionic constituents of honey as indicators of environmental pollution, botanical origin and authenticity.

Three beehive matrices, sampled in eighteen apiaries from West France, were analysed for the presence of lead (Pb). Samples were collected during four different periods in both 2008 and 2009. Honey was the matrix the least contaminated by Pb (min = 0.004 μg g⁻¹; max = 0.378 μg g⁻¹; mean = 0.047 μg g⁻¹; sd = 0.057). The contamination of bees (min = 0.001 μg g⁻¹; max = 1.869 μg g⁻¹; mean = 0.223 μg g⁻¹; sd = 0.217) and pollen (min = 0.004 μg g⁻¹; max = 0.798 μg g⁻¹; mean = 0.240 μg g⁻¹; sd = 0.200) showed similar levels and temporal variations but bees seemed to be more sensitive bringing out the peaks of Pb contamination. Apiaries in urban and hedgerow landscapes appeared more contaminated than apiaries in cultivated and island landscapes. Sampling period had a significant effect on Pb contamination with higher Pb concentrations determined in dry seasons.

Varroa mite is one of the major adverse factors causing honey bee population decline. In this study, Varroa destructor-infested and uninfested honey bee colonies were established by selective applying miticide (Apivar® amitraz). Mite population was monitored monthly (April–October 2016), and deformed wing virus (DWV) loading was detected seasonally (April, July, and October). Four immunity- and two physiology-related gene expressions, natural mortality, and susceptibility to five insecticides were comparatively and seasonally examined in field-collected honey bee workers. Results showed that Apivar-treated bee colonies had minor or undetectable mite and DWV (using RT-qPCR) infestations in whole bee season, while untreated colonies had substantially higher mite and DWV infestations. In untreated colonies, Varroa mite population irregularly fluctuated over the bee season with higher mite counts in Jun (318 ± 89 mites dropped in 48 h) or August (302) than that (25 ± 4 or 34) in October, and mite population density was not dynamically or closely correlated with the seasonal shift of honey bee natural mortality (regression slope = −0.5212). Unlike mite, DWV titer in untreated colonies progressively increased over the bee season, and it was highly correlated (R² = 1) with the seasonal increase of honey bee natural mortality. Significantly lower gene expressions of dor, PPO, mfe, potentially PPOa and eat as well, in untreated colonies also indicated an association of increased DWV infestation with decreased physiological and immunity-related functions in late-season honey bees. Furthermore, bees with lower mite/DWV infestations exhibited generally consistently lower susceptibilities (contact and oral toxicities) to five representative insecticides than the bees without Apivar treatment. All of these data from this study consistently indicated an interaction of Varroa/viral infestations with insecticide susceptibilities in honey bees, potentially through impairing bee's physiology and immunity, emphasizing the importance of mite control in order to minimize honey bee decline.

Commercial beekeepers in many locations are experiencing increased annual colony losses of honey bees (Apis mellifera), but the causes, including the role of agrochemicals in colony losses, remain unclear. In this study, we investigated the effects of chronic consumption of pollen containing a widely-used fungicide (Pristine®), known to inhibit bee mitochondria in vitro, which has recently been shown to reduce honey bee worker lifespan when field-colonies are provided with pollen containing field-realistic levels of Pristine®. We fed field colonies pollen with a field-realistic concentration of Pristine® (2.3 ppm) and a concentration two orders of magnitude higher (230 ppm). To challenge flight behavior and elicit near-maximal metabolic rate, we measured flight quality and metabolic rates of bees in two lower-than-normal air densities. Chronic consumption of 230 but not 2.3 ppm Pristine® reduced maximal flight performance and metabolic rates, suggesting that the observed decrease in lifespans of workers reared on field-realistic doses of Pristine®-laced pollen is not due to inhibition of flight muscle mitochondria. However, consumption of either the 230 or 2.3 ppm dose reduced thorax mass (but not body mass), providing the first evidence of morphological effects of Pristine®, and supporting the hypothesis that Pristine® reduces forager longevity by negatively impacting digestive or nutritional processes.

Mites are considered the worst enemy of honey bees, resulting in economic losses in agricultural production. In apiculture, flumethrin is frequently used to control mites. It causes residues of flumethrin in colonies which may threaten honey bees, especially for larvae. Still, the impact of flumethrin-induced dysbiosis on honey bees larval health has not been fully elucidated, and any impact of microbiota for decomposing flumethrin in honey bees is also poorly understood. In this study, 2-day-old larvae were fed with different flumethrin-sucrose solutions (0, 0.5, 5, 50 mg/kg) and the dose increased daily (1.5, 2, 2.5 and 3 μL) until capped, thereafter the expression level of two immune genes (hymenoptaecin, defensin1) and two detoxication-related genes (GST, catalase) were measured. Meanwhile, the effect of flumethrin on honey bee larvae (Apis mellifera) gut microbes was also explored via 16S rRNA Illumina deep sequencing. We found that flumethrin at 5 mg/kg triggered the over expression of immune-related genes in larvae, while the larval detoxification-related genes were up-regulated when the concentrations reached 50 mg/kg. Moreover, the abundance and diversity of microbes in flumethrin-treated groups (over 0.5 mg/kg) were significantly lower than control group, but it increased with flumethrin concentrations among the flumethrin-treated groups. Our results revealed that microbes served as a barrier in the honey bee gut and were able to protect honey bee larvae to a certain extent, and reduce the stress of flumethrin on honey bee larvae. In addition, as the concentration of flumethrin increases, honey bee larvae activate their immune system then detoxification system to defend against the potential threat of flumethrin. This is the first report on the impact of flumethrin on gut microbiota in honey bees larvae. The findings revealed new fundamental insights regarding immune and detoxification of host-associated microbiota.

Although fungicides were previously considered to be safe for important agricultural pollinators such as honey bees, recent evidence has shown that they can cause a number of behavioral and physiological sublethal effects. Here, we focus on the fungicide Pristine® (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin), which is sprayed during the blooming period on a variety of crops and is known to affect honey bee mitochondria at field-relevant levels. To date, no study has tested the effects of a field-relevant concentration of a fungicide on associative learning ability in honey bees. We tested whether chronic, colony-level exposure at field-relevant and higher concentrations of Pristine® impairs performance on the proboscis extension reflex (PER) paradigm, an associative learning task. Learning performance was reduced at higher field-relevant concentrations of Pristine®. The reductions in learning performance could not be explained by effects on hunger or motivation, as sucrose responsiveness was not affected by Pristine® exposure. To determine whether Pristine®‘s negative effects on learning performance were mediated at a specific life stage, we conducted a cross-fostering experiment that exposed bees to the fungicide either only as larvae, only as adults, or during both stages. We found that exposure across the entire life was necessary to significantly reduce learning performance, although non-significant reductions occurred when bees were exposed during just one stage. Our study provides strong evidence that Pristine® has significant sublethal effects on learning performance. As associative learning is a necessary ability for foraging, our results raise concerns that Pristine® could impair foraging abilities and substantially weaken colony health.