Translocation of Tebuconazole between Bee Matrices and Its Potential Threat on Honey Bee (Apis mellifera Linnaeus) Queens

Various pesticide residues can be found in different bee colony components. The queen larvae of honey bee (Apis mellifera L.) receive non-contaminated food from nurse bees. However, there is little knowledge about how pesticide residues affect developing bees. Additionally, little is known about the migration of lipophilic pesticides between bee matrices. While wax, royal jelly (RJ), and bee larvae are chemically distinct, they all contain lipids and we expected the lipophilic fungicide tebuconazole to be absorbed by different contacting materials. Our aim was to analyze the translocation of tebuconazole residues from queen cell wax to RJ, queen larvae, and newly emerged queens and to evaluate its potential risk to queens. We demonstrated the potential for the migration of tebuconazole from wax to RJ, with a strong dilution effect from the original contamination source. No residues were detected in queen bee larvae and newly emerged queens, indicating that the migration of tebuconazole probably did not directly endanger the queen bee, but there was some risk that tebuconazole might still affect the homeostasis of developing bees.

Seasonal Variations in the Organization and Structure of Apis cerana cerana Swarm Queen Cells

This paper describes the organization and structure of the swarm queen cells of Apis cerana cerana in spring, summer, and autumn in Kunming, Yunnan Province, China. We measured the following indices to reveal the organization rule of swarm cells: number of swarm cells built by each colony during different seasons; the shortest distance between two adjacent swarm cells on the comb; distance between swarm cell base and bottom bar of movable frame. We revealed the swarm cells structural characteristics using the following indicators: maximum diameter of swarm cell, the length between mouth and bottom of swarm cell, depth between maximum diameter and bottom of swarm cell, and the ratio of maximum diameter to depth between maximum diameter and bottom of swarm cell. Regarding seasonal differences, results indicated a significant variation in the distance between the swarm cell base and the bottom bar of the movable frame. Still, no such effect was observed in the shortest distance between two adjacent swarm cells. The maximum swarm cell diameter was not considerably influenced either, while the distance between the maximum diameter and the bottom of the swarm cell had substantial variation. The detected ratio of the maximum diameter to the depth between the maximum diameter and the bottom of theswarm cell indicated seasonal changes in the bottom shape of the swarm cell. This study clarifies the temporal and spatial distribution and structure of swarm cells of A. c. cerana. It establishes the basis for predicting the time and position of appearing swarm cells, thus allowing for a more precise determination of the shape and size of queen-cell punch and the ideal position of a cell cup on the bar of queen cup frames in artificial queen rearing.

Effect of different queen cell sizes on the acceptance of grafted larvae in Apis cerana javana Fabr. Rearing

This study aims to evaluate Apis cerana javana Fabr. rearing based on differences in the queen cup sizes. A total of four queen bees were used with three different treatments and six replications, meanwhile, this study was conducted at Kembang Joyo Bee Farm, Malang, East Java. The results showed that the differences in the queen cup size had no significant effect on the success rate of queen rearing (p>0.05). The highest percentage of larvae acceptance (79%) was found in P3 treatment, followed by P2 (71%), while the lowest was found in P1 (63%). Furthermore, the highest percentage of larvae to pupae metamorphosis was found in P1 and P2 with a success rate of 100%, while the lowest was found in P3 with 89%. The highest percentage of pupae emergence was found in treatment P1 and P2 with a success rate of 100% and P3 with 94%. Based on the results, the difference queen cup sizes had no significant effect on the success rate of Apis cerana javana Fabr. rearing.

Nosema Ceranae Interactions with Nosema apis and Black Queen Cell Virus

Nosema ceranae is a relatively new pathogen of the honeybee (Apis mellifera) and the course of type C nosemosis (the disease that it causes) is not entirely known. In order to better understand the course and the consequences of this disease, laboratory experiments were performed. They aimed to compare the course of N. ceranae infection with the course of Nosema apis infection, taking its influence on the black queen cell virus (BQCV) into account. Determination of the quantity of N. ceranae and BQCV genetic material in laboratory tests was performed using real-time PCR. In mixed Nosema infections, N. ceranae “wins” the competition and manages to outnumber N. apis significantly. BQCV exacerbates the course of both A and C nosemoses, but the data shows that in the case of nosemosis C and this viral infection, the mortality rate was the highest from all examined groups. Obtained results show that N. ceranae is more pathogenic for A. mellifera than N. apis, and the course of type C nosemosis is much heavier, which results in the shortened life spans of bees, and in connection with BQCV it becomes even more dangerous to bees.

An Investigation of Honey Bee Viruses Prevalence in Managed Honey Bees (Apis mellifera and Apis cerana) Undergone Colony Decline

Objective: In the absence of known clinical symptoms, viruses were considered to be the most probable key pathogens of honey bee. Therefore, the aim of this study was to investigate the prevalence and distribution of honey bee viruses in managed Apis mellifera and Apis cerana in China. Methods: We conducted a screening of 8 honey bee viruses on A. mellifera and A. cerana samples collected from 54 apiaries from 13 provinces in China using RT-PCR. Results: We found that the types and numbers of viral species significantly differed between A. mellifera and A. cerana. Black Queen Cell Virus (BQCV), Chronic Bee Paralysis Virus (CBPV), Apis mellifera filamentous virus (AmFV), and Kakugo virus (DWV-A/KV) were the primary viruses found in A. mellifera colonies, whereas Chinese Sacbrood Bee Virus (CSBV) and Sacbrood Bee Virus (SBV) were the primary viruses found in A. cerana. The percentage infection of BQCV and CSBV were 84.6% and 61.6% in all detected samples. We first detected the occurrences of Varroa destructor virus-1 (VDV-1 or DWV-B) and DWV-A/KV in China but not ABPV in both A. mellifera and A. cerana. Conclusion: This study showed that BQCV and CSBV are the major threat to investigated A. mellifera and A. cerana colonies.

The Pathogens Spillover and Incidence Correlation in Bumblebees and Honeybees in Slovenia

Slovenia has a long tradition of beekeeping and a high density of honeybee colonies, but less is known about bumblebees and their pathogens. Therefore, a study was conducted to define the incidence and prevalence of pathogens in bumblebees and to determine whether there are links between infections in bumblebees and honeybees. In 2017 and 2018, clinically healthy workers of bumblebees (Bombus spp.) and honeybees (Apis mellifera) were collected on flowers at four different locations in Slovenia. In addition, bumblebee queens were also collected in 2018. Several pathogens were detected in the bumblebee workers using PCR and RT-PCR methods: 8.8% on acute bee paralysis virus (ABPV), 58.5% on black queen cell virus (BQCV), 6.8% on deformed wing virus (DWV), 24.5% on sacbrood bee virus (SBV), 15.6% on Lake Sinai virus (LSV), 16.3% on Nosema bombi, 8.2% on Nosema ceranae, 15.0% on Apicystis bombi and 17.0% on Crithidia bombi. In bumblebee queens, only the presence of BQCV, A. bombi and C. bombi was detected with 73.3, 26.3 and 33.3% positive samples, respectively. This study confirmed that several pathogens are regularly detected in both bumblebees and honeybees. Further studies on the pathogen transmission routes are required.

The Comparison of Honeybee Viral Loads for Six Honeybee Viruses (ABPV, BQCV, CBPV, DWV, LSV3 and SBV) in Healthy and Clinically Affected Honeybees with TaqMan Quantitative Real-Time RT-PCR Assays

The viral loads of acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV), Lake Sinai virus 3 (LSV3), and sacbrood bee virus (SBV) were determined in samples with the use of quantitative TaqMan real-time reverse transcription and polymerase chain reaction (RT-qPCR). A total of 108 samples of healthy adult honeybees from four differently located apiaries and samples of honeybees showing different clinical signs of viral infections from 89 apiaries were collected throughout Slovenia. The aim of this study was to discover correlations between viral loads and clinical signs in adult honeybees and confirm previously set threshold viral load levels between healthy and clinically affected honeybees. Within this study, two new RT-qPCR assays for quantification of LSV3 and SBV were developed. Statistically significant differences in viral loads of positive samples were identified between healthy and clinically affected honeybees for ABPV, CBPV, DWV, and SBV, while for BQCV and LSV3, no statistical differences were observed between both groups. Despite high detected LSV3 prevalence and viral loads around 6.00 log10 viral copies/bee, this lineage probably has a limited impact on the health status of honeybee colonies. The determined viral loads between 3.94 log10 and 13.17 log10 in positive samples for six viruses, collected over 10 consecutive months, including winter, present additional information of high viral load variations in healthy honeybee colonies.

The First Detection and Genetic Characterization of Four Different Honeybee Viruses in Wild Bumblebees from Croatia

To determine the presence and the prevalence of four different honeybee viruses (acute bee paralysis virus—ABPV, black queen cell virus—BQCV, chronic bee paralysis virus—CBPV, deformed wing virus—DWV) in wild bumblebees, pooled randomly selected bumblebee samples were collected from twenty-seven different locations in the territory of Croatia. All samples were prepared and examined using the RT-PCR methods for quantification of mentioned honeybee viruses. Determined prevalence (%) of identified positive viruses were in the following decreasing order: BQCV > DWV > ABPV, CBPV. Additionally, direct sequencing of samples positive for BQCV (n = 24) and DWV (n = 2) was performed, as well as a test of molecular phylogeny comparison with those available in GenBank. Selected positive field viruses’ strains showed 95.7 to 100% (BQCV) and 98.09% (DWV) nucleotide identity with previously detected and deposited honeybee virus strains in the geographic areas in Croatia and neighboring Slovenia. In this article, the first detection of four honeybee viruses with genetic characterization of high diversity strains circulating in wild bumblebees in Croatia is presented.

Feral and managed honey bees, Apis mellifera (Hymenoptera: Apidae), in southern California have similar levels of viral pathogens

Bees provide critical pollination services but are threatened by multiple stressors, including viral pathogens. Most studies of pollinator health focus on managed honey bees (Apis mellifera Linnaeus) (MHB) or native bee species, but a third player, the feral honey bee (FHB), requires further study. Spillover and spillback of viral pathogens between these managed, feral, and native bees is generating increasing interest. In this case study, we provide evidence suggesting that FHB colonies play an important role in viral pathogen dynamics of southern California pollinator communities because they act as reservoirs, of viral pathogens such as acute bee paralysis virus (ABPV), black queen cell virus (BQCV), and deformed wing virus (DWV). Surprisingly, even though FHB are not treated for diseases or parasites, they harbor similar pathogen loads to MHB, which are usually highly treated, suggesting the need for future studies to determine if FHB resist or are more resilient to viruses.

Parasites and RNA viruses in wild and laboratory reared bumble bees Bombus pauloensis (Hymenoptera: Apidae) from Uruguay

Bumble bees (Bombus spp.) are important pollinators insects involved in the maintenance of natural ecosystems and food production. Bombus pauloensis is a widely distributed species in South America, that recently began to be managed and commercialized in this region. The movement of colonies within or between countries may favor the dissemination of parasites and pathogens, putting into risk while populations of B. pauloensis and other native species. In this study, wild B. pauloensis queens and workers, and laboratory reared workers were screened for the presence of phoretic mites, internal parasites (microsporidia, protists, nematodes and parasitoids) and RNA viruses (Black queen cell virus (BQCV), Deformed wing virus (DWV), Acute paralysis virus (ABCV) and Sacbrood virus (SBV)). Bumble bee queens showed the highest number of mite species, and it was the only group where Conopidae and S. bombi were detected. In the case of microsporidia, a higher prevalence of N. ceranae was detected in field workers. Finally, the bumble bees presented the four RNA viruses studied for A. mellifera, in proportions similar to those previously reported in this species. Those results highlight the risks of spillover among the different species of pollinators.