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.

Antibiotics are omnipresent in the environment due to their widespread use, and they have wide-ranging negative impacts on organisms. Virus resistance differs substantially between domesticated Apis mellifera and wild Apis cerana, although both are commonly raised in China. Here, we investigated whether antibiotics can increase the sensitivity of honey bees to viral infection using the Israeli acute paralysis virus (IAPV) and tetracycline as representative virus and antibiotic. Although IAPV multiplied to lower levels in A. cerana than A. mellifera, resulting in decreased mortality (P < 0.01), there was no significant difference in immune responses to viral infection between the two species. Adult worker bees (A. cerana and A. mellifera) were treated with or without tetracycline to demonstrate the prominent role of gut microbiota against viral infection, and found Lactobacillus played a vital antiviral role in A. cerana. In A. cerana but not A. mellifera, tetracycline treatment reduced clearly bee survival and increased susceptibility to IAPV infection (P < 0.01). Our findings revealed that long-term antibiotic treatment in A. mellifera had altered the native gut microbiome and promoted the sensitivity to viral infection. We highlight the effects of antibiotics exposure on resistance to microbial and viral infection.

Honey bees (Apis mellifera) and other pollinator populations are declining worldwide, and the reasons remain controversial. Based on laboratory testing, fungicides have traditionally been considered bee-safe. However, there have been no experimental tests of the effects of fungicides on colony health under field conditions, and limited correlational data suggests there may be negative impacts on bees at levels experienced in the field. We tested the effects of one of the most commonly used fungicides on colony health by feeding honey bee colonies pollen containing Pristine® (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin) at four levels that bracketed concentrations we measured for pollen collected by bees in almond orchards. We also developed a method for calculating per-bee and per-larva dose. Pristine® consumption significantly and dose-dependently reduced worker lifespan and colony population size, with negative health effects observed even at the lowest doses. The lowest concentration we tested caused a 15% reduction in the worker population at an estimated dosage that was three orders of magnitude below the estimated LD₁₅ values for previous acute laboratory studies. The enhanced toxicity under field conditions is at least partially due to activation of colonial nutritional responses missed by lab tests. Pristine® causes colonies to respond to perceived protein malnutrition by increasing colony pollen collection. Additionally, Pristine induces much earlier transitioning to foraging in individual workers, which could be the cause of shortened lifespans. These findings demonstrate that Pristine® can negatively impact honey bee individual and colony health at concentrations relevant to what they experience from pollination behavior under current agricultural conditions.

Due to their extensive use in both agricultural and non-agricultural applications, pesticides are a major source of environmental contamination. Honey bee colonies are proven sentinels of these and other contaminants, as they come into contact with them during their foraging activities. However, active sampling strategies involve a negative impact on these organisms and, in most cases, the need of analyzing multiple heterogeneous matrices. Conversely, the APIStrip-based passive sampling is innocuous for the bees and allows for long-term monitorings using the same colony. The versatility of the sorbent Tenax, included in the APIStrip composition, ensures that comprehensive information regarding the contaminants inside the beehive will be obtained in one single matrix. In the present study, 180 APIStrips were placed in nine apiaries distributed in Denmark throughout a six-month sampling period (10 subsequent samplings, April to September 2020). Seventy-five pesticide residues were detected (out of a 428-pesticide scope), boscalid and azoxystrobin being the most frequently detected compounds. There were significant variations in the findings of the sampling sites in terms of number of detections, pesticide diversity and average concentration. A relative indicator of the potential risk of pesticide exposure for the honey bees was calculated for each sampling site. The evolution of pesticide detections over the sampling periods, as well as the individual tendencies of selected pesticides, is herein described. The findings of this large-scale monitoring were compared to the ones obtained in a previous Danish, APIStrip-based pilot monitoring program in 2019. Samples of honey and wax were also analyzed and compared to the APIStrip findings.

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.

Thiacloprid is widely used in agriculture and may affect pollinators. However, its molecular effects are poorly known. Here, we report the global gene expression profile in the brain of honey bee foragers assessed by RNA-sequencing. Bees were exposed for 72 h to nominal concentrations of 25 and 250 ng/bee via sucrose solution. Determined residue concentrations by LC-MS/MS were 0.59 and 5.49 ng/bee, respectively. Thiacloprid exposure led to 5 and 71 differentially expressed genes (DEGs), respectively. Nuclear genes encoding mitochondrial ribosomal proteins and enzymes involved in oxidative phosphorylation, as well as metabolism enzymes and transporters were altered at 5.49 ng/bee. Kyoto Encylopedia of Genes and Genomes (KEGG) analysis revealed that mitochondrial ribosome proteins, mitochondrial oxidative phosphorylation, pyrimidine, nicotinate and nicotinamide metabolism and additional metabolic pathways were altered. Among 21 genes assessed by RT-qPCR, the transcript of farnesol dehydrogenase involved in juvenile hormone III synthesis was significantly down-regulated. Transcripts of cyp6a14-like and apolipophorin-II like protein, cytochrome oxidase (cox17) and the non-coding RNA (LOC102654625) were significantly up-regulated at 5.49 ng/bee. Our findings indicate that thiacloprid causes transcriptional changes of genes prominently associated with mitochondria, particularly oxidative phosphorylation. This highlight potential effects of this neonicotinoid on energy metabolism, which may compromise bee foraging and thriving populations at environmentally relevant concentrations.

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.

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.

Honey bees Apis mellifera forage in a wide radius around their colony, bringing back contaminated food resources that can function as terrestrial bioindicators of environmental pesticide exposure. Evaluating pesticide exposure risk to pollinators is an ongoing problem. Here we apply five metrics for pesticide exposure risk (prevalence, diversity, concentration, significant pesticide prevalence, and hazard quotient (HQ)) to a nation-wide field study of honey bees, Apis mellifera in the United States. We examined samples from 1055 apiaries over seven years for 218 different pesticide residues and metabolites, determining that bees were exposed to 120 different pesticide products with a mean of 2.78 per sample. Pesticides in pollen were highly prevalent and variable across states. While pesticide diversity increased over time, most detections occurred at levels predicted to be of low risk to colonies. Varroacides contributed most to concentration, followed by fungicides, while insecticides contributed most to diversity above a toxicity threshold. High risk samples contained one of 12 different insecticides or varroacides. Exposures predicted to be low-risk were nevertheless associated with colony morbidity, and low-level fungicide exposures were tied to queen loss, Nosema infection, and brood diseases.