The Improving Effects of Probiotic-Added Pollen Substitute Diets on the Gut Microbiota and Individual Health of Honey Bee (Apis mellifera L.).

Honey bee (Apis mellifera L.) health is crucial for honey bee products and effective pollination, and it is closely associated with gut bacteria. Various factors such as reduced habitat, temperature, disease, and diet affect the health of honey bees by disturbing the homeostasis of the gut microbiota. In this study, high-throughput 16S rRNA gene sequencing was used to analyze the gut microbiota of honey bees subjected to seven diets over 5 days. Lactobacillus dominated the microbiota in all diets. Cage experiments (consumption, head protein content, and vitellogenin gene expression level) were conducted to verify the effect of the diet. Through a heatmap, the Diet2 (probiotic-supplemented) group was clustered together with the Beebread and honey group, showing high consumption (177.50 ± 26.16 mg/bee), moderately higher survival duration (29.00 ± 2.83 days), protein content in the head (312.62 ± 28.71 µg/mL), and diet digestibility (48.41 ± 1.90%). Additionally, we analyzed the correlation between gut microbiota and health-related indicators in honey bees fed each diet. Based on the overall results, we identified that probiotic-supplemented diets increased gut microbiota diversity and positively affected the overall health of individual honey bees.

Microfungal flora of Apis mellifera anatoliaca (Hymenoptera: Apidae) and Varroa destructor (Mesostigmata: Varroidae) from the Eastern Black Sea Region and fungal vector capacity in honey bee colonies.

Honey bees have a great economic importance both in Turkey and in the world due to the products they produce and their contribution to pollination. For this reason, many microflora and microbiota studies have been conducted on bees. While these research were primarily focused on pathogen isolation, the ecological roles of non-pathogenic flora members and how they may be used are now being studied more extensively. Considering the importance of pathogens, the number of studies is expected to continue to increase. This study was carried out to determine the microfungal flora of the body surfaces and digestive tracts of dead honey bee (Apis mellifera anatoliaca) and Varroa destructor samples taken from different apiaries in the Eastern Black Sea Region of Turkey (Gümüshane, Trabzon, Artvin and Ordu) in 2022. As a result of the study, a total of 11 different fungal species belonging to the genera Penicillium, Alternaria, Mucor, Trichoderma, Fusarium, Aspergillus and Verticillium were identified and the relationships of these fungi with bees were discussed based on the literature.

The oriental hornet (Vespa orientalis) as a potential vector of honey bee's pathogens and a threat for public health in North-East Italy.

BACKGROUND: Oriental hornets are large predatory hymenoptera that occur in the southern part of Asia and the southeastern Mediterranean. Among many pests of bee colonies, Vespa orientalis was recorded to be one of the most destructive. OBJECTIVES: The aim of this study was to: (1) monitor the presence of pathogens carried by V. orientalis that could potentially threaten honey bees and public health; (2) describe the hornet's predatory behavior on honey bee colonies and (3) collect the medical history of a V. orientalis sting suffered by a 36-year-old woman. METHODS: Observations of V. orientalis predatory behavior and the catches of hornets for parasitological and microbiological examination, using molecular and bacteriological analyses, were carried out in three experimental apiaries, both in spring in order to capture the foundress queens and during the summer to capture the workers. Furthermore, the medical history and photographic documentation of a V. orientalis sting suffered by a 36-year-old woman have been collected. RESULTS: The results obtained highlight that V. orientalis is capable of causing serious damage to beekeeping by killing bees, putting under stress the honey bee colonies and by potentially spreading honey bee pathogens among apiaries. These hornets may also become a public health concern, since they are capable of inflicting multiple, painful stings on humans. CONCLUSIONS: Only the development of an Integrated Management Control Program will be able to contain the negative effects of anomalous population growth and the potentially negative impact on honey bees and public health of V. orientalis.

New reference genomes of honey bee-associated bacteria Paenibacillus melissococcoides, Paenibacillus dendritiformis, and Paenibacillus thiaminolyticus.

We sequenced the genomes of recently discovered Paenibacillus melissococcoides (CCOS 2000) and of the type strains of closely related P. thiaminolyticus (DSM 7262) and P. dendritiformis (LMG 21716). The three genomes set the basis to unambiguous diagnostic of these honey bee associated Paenibacillus bacteria.

Survey Results of Honey Bee Colony Losses in Winter in China (2009-2021).

There is growing concern that massive loss of honey bees can cause serious negative effects on biodiversity and ecosystems. Surveys of colony losses have been performed worldwide to monitor the dynamic changes and health status of honey bee colonies. Here, we present the results of surveys regarding winter colony losses from 21 provinces in China from 2009 to 2021, with a total of 1,744,324 colonies managed by 13,704 beekeepers. The total colony losses were low (9.84%; 95% Confidence Interval (CI): 9.60-10.08%) but varied among years, provinces, and scales of apiaries. As little is known about the overwintering mortality of Apis cerana, in this study, we surveyed and compared the loss rates between Apis mellifera and A. cerana in China. We found colonies of A. mellifera suffered significantly lower losses than A. cerana in China. Larger apiaries resulted in higher losses in A. mellifera, whereas the opposite was observed in A. cerana. Furthermore, we used generalized linear mixed-effects models (GLMMs) to evaluate the effects of potential risk factors on winter colony losses and found that the operation size, species, migration, migration×species interaction, and queen problems were significantly related to the loss rates. New queens can increase their colony overwintering survival. Migratory beekeepers and large operations reported lower loss rates.

Building composite indices in the age of big data - Application to honey bee exposure to infectious and parasitic agents.

Pollinator insects play a crucial role in maintaining biodiversity and agricultural production worldwide. Yet they are subject to various infectious and parasitic agents (IPAs). To better assess their exposure to IPAs, discriminative and quantitative molecular methods have been developed. These tools produce large datasets that need to be summarised so as to be interpreted. In this paper, we described the calculation of three types of composite indices (numerical, ordinal, nominal) to characterize the honey bee exposure to IPAs in 128 European sites. Our summarizing methods are based on component-based factorial analyses. The indices summarised the dataset of eight IPAs quantified at two sampling times, into synthetic values providing different yet complementary information. Because our dataset included two sampling times, we used Multiple Factor Analysis (MFA) to synthetize the information. More precisely, the numerical and ordinal indices were generated from the first component of MFA, whereas the nominal index used the first main components of MFA combined with a clustering analysis (Hierarchical Clustering on components). The numerical index was easy to calculate and to be used in further statistical analyses. However, it contained only about 20% of the original information. Containing the same amount of original information, the ordinal index was much easier to interpret. These two indices summarised information in a unidimensional manner. Instead, the nominal index summarised information in a multidimensional manner, which retained much more information (94%). In the practical example, the three indices showed an antagonistic relationship between N. ceranae and DWV-B. These indices represented a toolbox where scientists could pick one composite index according to the aim pursued. Indices could be used in further statistical analyses but could also be used by policy makers and public instances to characterize a given sanitary situation at a site level for instance.

Biocontrol potential of Apilactobacillus kunkeei EIR/BG-1 against infectious diseases in honey bees (Apis mellifera L.).

The significant reduction of honey bee colonies due to the various infectious agents highlights the need for the development of new alternatives and integrated management strategies to keep a hive strong and healthy. The main purpose of this study was to develop an environmentally and friendly microbial feed supplements to prevent honey bee mortality and keep the bee colony population healthy and productive. For this aim, Apilactobacillus kunkeei EIR/BG-1 isolated from honey bee gut microbiota was evaluated for its preventive role against American Foulbrood disease and nosemosis. To test the ability of the strain EIR/BG-1 for suppressing Paenibacillus larvae growth under in vitro conditions, the agar well diffusion method was used and viable cells of the strain EIR/BG-1 inhibited the growth of P. larvae with an efficient inhibition zone (24 ± 0.8 mm) similar to tetracycline antibiotic (30 µg). To determine the preventive role of the strain EIR/BG-1 on infection progression, its viable cells were applied against nosemosis in a laboratory experimental setting. Our results showed that prophylactic supplementation of Al. kunkeei EIR/BG-1 (106 cfu/bee) significantly reduced the spore load (66 ± 6.1%). Besides, gene expression of antimicrobial peptides in gut tissue has been up-regulated and infected midgut epithelium integrity and peritrophic membrane production were improved. In conclusion, our findings suggest that prophylactic supplementation of Al. kunkeei EIR/BG-1 as a natural strategy may enhance the honey bee's response when challenged by pathogens. Field applications towards gaining a better understanding of its biocontrol role will be the main goal of our future researches.

Honey bee (Apis mellifera jemenitica) colony performance and queen fecundity in response to different nutritional practices.

Honey bee colony nutritional dynamics depend on the availability of floral resources throughout a countryside with varying forage circumstances. Few studies quantify the queen fecundity and colony performance about certain management approaches on a broad scale. The present study was conducted to investigate the queen bee fecundity and various colony performance parameters in response to different nutritional practices, i.e., Group-I, supplied with sucrose solution (1:1; w/v), Group-II, provided with locally available commercial pollen substitute, Group-III, supplied with both sucrose solution + locally available commercial pollen substitute, and Group-IV without any sugar solution and pollen substitute. Our results demonstrated that eggs laid by queen bees were significantly higher (1241.83 ± 46.24) in Group-III than in other groups over the time of observations. Similarly, a significant difference was noticed in the mean sealed worker brood area and honey store area between the different groups of management practices. Both, the max mean sealed worker brood area (2153.53 ± 29.18 cm2) and max mean honey store area (1713.33 ± 12.06 cm2) were observed in Group-III while, the mini mean sealed worker brood area (1066.53 ± 20.18 cm2) and mini mean honey store area (1058.86 ± 4.07 cm2) were observed in Group-IV. In contrast, a non-significant difference was observed in pollen stores between Group-II and Group-III (p > 0.005). Current findings add to our understanding of the mechanisms that underpin large-scale controlled colony performance when the natural pollens resources are insufficient.

Detection and Concentration of Neonicotinoids and Other Pesticides in Honey from Honey Bee Colonies Located in Regions That Differ in Agricultural Practices: Implications for Human and Bee Health.

This is a preliminary study conducted to analyze the presence and concentration of pesticides in honey obtained from honey bee colonies located in two regions with managed ecosystems that differ in the intensity and technification of agricultural practices. Fourteen pesticides at variable concentrations were detected in 63% of the samples analyzed. The pesticides most frequently found at higher concentrations were insecticides (neonicotinoids, followed by organophosphates), herbicides, and fungicides. The number, frequency, and concentration of pesticides were higher in samples collected from hives located where intensive and highly-technified agriculture is practiced. Forty-three percent of the samples from that zone had residues of imidacloprid, compared with only 13% of the samples from the less-technified zone. Furthermore, 87.5% of those samples had imidacloprid concentrations that were above sublethal doses for honey bees (>0.25 ng/g) but that are not considered hazardous to human health by the European Commission. The results of this study suggest that honey can be used as a bioindicator of environmental contamination by pesticides, which highlights the need to continue monitoring contaminants in this product to determine the risks of pesticide impacts on pollinator health, on ecosystems, and on their potential implications to human health and other non-target organisms.

Larval Exposure to Parasitic Varroa destructor Mites Triggers Specific Immune Responses in Different Honey Bee Castes and Species.

Innate immune systems are key defenses of animals and particularly important in species that lack the sophisticated adaptive immune systems as found in vertebrates. Here, we were interested to quantify variation in innate immune responses of insects in hosts that differ in their parasite susceptibility. To do this, we studied immune responses in honey bees, which can host a remarkable number of different parasites, which are major contributors of declining bee health and colony losses. The most significant parasite of honey bees is the mite Varroa destructor, which has infested the majority of global honey bee populations, and its control remains a major challenge for beekeepers. However, a number of nonmanaged honey bees seem able to control Varroa infections, for example, the Eastern honey bee Apis ceranacerana or the African honey bee Apis mellifera scutellata. These bees therefore make interesting study subjects to identify underlaying resistance traits, for example, by comparing them to more susceptible bee genotypes such as Western honey bees (A. melliferaligustica). We conducted a series of interlinked experiments and started with behavioral assays to compare the attractiveness of bee larvae to mites using different honey bee genotypes and castes. We found that 6-day-old larvae are always most attractive to mites, independently of genotype or castes. In a next step, we compared volatile profiles of the most attractive larvae to test whether they could be used by mites for host selection. We found that the abundance of volatile compounds differed between larval ages, but we also found significant differences between genotypes and castes. To further study the expected underlaying physiological differences between potentially resistant and susceptible host larvae, we compared the larval hemolymph proteomes of the three honey bee genotypes and two castes in response to mite exposure. We identified consistent upregulation of immune and stress-related genes in Varroa-exposed larvae, which differed between genotypes and castes. Tolerant honey bee castes and genotypes were characterized by stronger or more distinct immune esponses. In summary, we provide first insights into the complex involvement of the innate immune system of tolerant honey bees against mite infestations, which could be used for future breeding purposes.

Honey Bees and Industrial Agriculture: What Researchers are Missing, and Why it's a Problem.

Industrial agriculture is the root cause of many health problems that honey bees (Apis mellifera Linneaus, 1758) face, but honey bee researchers seldom call attention to this fact. We often discuss the stressors that contribute to colony loss (e.g., pathogens, pesticides, poor nutrition), but we rarely talk about where those stressors come from. This is a problem because we cannot resolve honey bee health issues unless we confront the systems that cause them harm. In this forum article, I unpack the relationship between honey bee health and industrial agriculture. I propose steps we can take to reframe our research to account for the impacts of this destructive system, and I discuss the uncomfortable questions that surface when we engage in this process. The goal of this article is to encourage conversation within the honey bee research community around the impacts of industrial agriculture, so that we can fully engage in the transformative change needed to support honey bee health.

Epidemiology and Biosecurity for Veterinarians Working with Honey bees (Apis mellifera).

Honeybee veterinary medicine is a developing field in Canada and the United States. Veterinarians interested in working with honeybees should develop a comprehensive knowledge base on disease dynamics as it applies to the individual, colony, apiary, and broader honeybee populations. There are currently several governmental, academic, and industry organizations that are carrying out epidemiological-based surveys. Although honeybees face unique challenges in regard to biosecurity, the basic principles still apply. Veterinarians can use their expertise in the area of biosecurity to make improvements to current protocols within the apiary and beekeeping operations.

Impact of Chronic Exposure to Sublethal Doses of Glyphosate on Honey Bee Immunity, Gut Microbiota and Infection by Pathogens.

Glyphosate is the most used pesticide around the world. Although different studies have evidenced its negative effect on honey bees, including detrimental impacts on behavior, cognitive, sensory and developmental abilities, its use continues to grow. Recent studies have shown that it also alters the composition of the honey bee gut microbiota. In this study we explored the impact of chronic exposure to sublethal doses of glyphosate on the honey bee gut microbiota and its effects on the immune response, infection by Nosema ceranae and Deformed wing virus (DWV) and honey bee survival. Glyphosate combined with N. ceranae infection altered the structure and composition of the honey bee gut microbiota, for example by decreasing the relative abundance of the core members Snodgrassella alvi and Lactobacillus apis. Glyphosate increased the expression of some immune genes, possibly representing a physiological response to mitigate its negative effects. However, this response was not sufficient to maintain honey bee health, as glyphosate promoted the replication of DWV and decreased the expression of vitellogenin, which were accompanied by a reduced life span. Infection by N. ceranae also alters honey bee immunity although no synergistic effect with glyphosate was observed. These results corroborate previous findings suggesting deleterious effects of widespread use of glyphosate on honey bee health, and they contribute to elucidate the physiological mechanisms underlying a global decline of pollination services.

Prioritizing changes in management practices associated with reduced winter honey bee colony losses for US beekeepers.

Beekeepers attempt to manage their honey bee colonies in ways that optimize colony health. Disentangling the impact of management from other variables affecting colony health is complicated by the diversity of practices used and difficulties handling typically complex and incomplete observational datasets. We propose a method to 1) compress multi-factored management data into a single index, to holistically investigate the real world impact of management on colony mortality, and 2) simplify said index to identify the core practices for which a change in behavior is associated with the greatest improvement in survivorship. Experts scored the practices of US beekeepers (n = 18,971) documented using four years of retrospective surveys (2012-2015). Management Index scores significantly correlated with loss rates, with beekeepers most in line with recommendations suffering lower losses. The highest ranked practices varied by operation type, as recommendations accounted for the current prevalence of practices. These results validate experts' opinion using empirical data, and can help prioritize extension messages. Improving management will not prevent all losses; however, we show that few behavioral changes (in particular related to comb management, sources of new colonies and Varroa management) can lead to a non-negligible reduction in risk.

Chronic High Glyphosate Exposure Delays Individual Worker Bee (Apis mellifera L.) Development under Field Conditions.

The ongoing debate about glyphosate-based herbicides (GBH) and their implications for beneficial arthropods gives rise to controversy. This research was carried out to cover possible sublethal GBH effects on the brood and colony development, adult survival, and overwintering success of honey bees (Apis mellifera L.) under field conditions. Residues in bee relevant matrices, such as nectar, pollen, and plants, were additionally measured. To address these questions, we adopted four independent study approaches. For brood effects and survival, we orally exposed mini-hives housed in the "Kieler mating-nuc" system to sublethal concentrations of 4.8 mg glyphosate/kg (T1, low) and 137.6 mg glyphosate/kg (T2, high) over a period of one brood cycle (21 days). Brood development and colony conditions were assessed after a modified OECD method (No. 75). For adult survival, we weighed and labeled freshly emerged workers from control and exposed colonies and introduced them into non-contaminated mini-hives to monitor their life span for 25 consecutive days. The results from these experiments showed a trivial effect of GBH on colony conditions and the survival of individual workers, even though the hatching weight was reduced in T2. The brood termination rate (BTR) in the T2 treatment, however, was more than doubled (49.84%) when compared to the control (22.11%) or T1 (20.69%). This was surprising as T2 colonies gained similar weight and similar numbers of bees per colony compared to the control, indicating an equal performance. Obviously, the brood development in T2 was not "terminated" as expected by the OECD method terminology, but rather "slowed down" for an unknown period of time. In light of these findings, we suggest that chronic high GBH exposure is capable of significantly delaying worker brood development, while no further detrimental effects seem to appear at the colony level. Against this background, we discuss additional results and possible consequences of GBH for honey bee health.

Using Manual and Computer-Based Text-Mining to Uncover Research Trends for Apis mellifera.

Honey bee research is believed to be influenced dramatically by colony collapse disorder (CCD) and the sequenced genome release in 2006, but this assertion has never been tested. By employing text-mining approaches, research trends were tested by analyzing over 14,000 publications during the period of 1957 to 2017. Quantitatively, the data revealed an exponential growth until 2010 when the number of articles published per year ceased following the trend. Analysis of author-assigned keywords revealed that changes in keywords occurred roughly every decade with the most fundamental change in 1991-1992, instead of 2006. This change might be due to several factors including the research intensification on the Varroa mite. The genome release and CCD had quantitively only minor effects, mainly on honey bee health-related topics post-2006. Further analysis revealed that computational topic modeling can provide potentially hidden information and connections between some topics that might be ignored in author-assigned keywords.

Diversity and Global Distribution of Viruses of the Western Honey Bee, Apis mellifera.

In the past centuries, viruses have benefited from globalization to spread across the globe, infecting new host species and populations. A growing number of viruses have been documented in the western honey bee, Apis mellifera. Several of these contribute significantly to honey bee colony losses. This review synthetizes the knowledge of the diversity and distribution of honey-bee-infecting viruses, including recent data from high-throughput sequencing (HTS). After presenting the diversity of viruses and their corresponding symptoms, we surveyed the scientific literature for the prevalence of these pathogens across the globe. The geographical distribution shows that the most prevalent viruses (deformed wing virus, sacbrood virus, black queen cell virus and acute paralysis complex) are also the most widely distributed. We discuss the ecological drivers that influence the distribution of these pathogens in worldwide honey bee populations. Besides the natural transmission routes and the resulting temporal dynamics, global trade contributes to their dissemination. As recent evidence shows that these viruses are often multihost pathogens, their spread is a risk for both the beekeeping industry and the pollination services provided by managed and wild pollinators.

Propolis Consumption Reduces Nosema ceranae Infection of European Honey Bees (Apis mellifera).

Nosema ceranae is a widespread obligate intracellular parasite of the ventriculus of many species of honey bee (Apis), including the Western honey bee Apis mellifera, in which it may lead to colony death. It can be controlled in A. mellifera by feeding the antibiotic fumagillin to a colony, though this product is toxic to humans and its use has now been banned in many countries, so in beekeeping, there exists a need for alternative and safe products effective against N. ceranae. Honeybees produce propolis from resinous substances collected from plants and use it to protect their nest from parasites and pathogens; propolis is thought to decrease the microbial load of the hive. We hypothesized that propolis might also reduce N. ceranae infection of individual bees and that they might consume propolis as a form of self-medication. To test these hypotheses, we evaluated the effects of an ethanolic extract of propolis administered orally on the longevity and spore load of experimentally N. ceranae-infected worker bees and also tested whether infected bees were more attracted to, and consumed a greater proportion of, a diet containing propolis in comparison to uninfected bees. Propolis extracts and ethanol (solvent control) increased the lifespan of N. ceranae-infected bees, but only propolis extract significantly reduced spore load. Our propolis extract primarily contained derivatives of caffeic acid, ferulic acid, ellagic acid and quercetin. Choice, scan sampling and food consumption tests did not reveal any preference of N. ceranae-infected bees for commercial candy containing propolis. Our research supports the hypothesis that propolis represents an effective and safe product to control N. ceranae but worker bees seem not to use it to self-medicate when infected with this pathogen.

Hazard of a neonicotinoid insecticide on the homing flight of the honeybee depends on climatic conditions and Varroa infestation.

The paradigm for all toxicological bioassays in the risk assessment of pesticide registration reflects the principle that experimental conditions should be controlled to avoid any other factors that may affect the endpoint measures. As honeybee colonies can be frequently exposed to bio-aggressors in real conditions, often concomitantly with pesticides, co-exposure to pesticide/bio-aggressors is becoming a concern for regulatory authorities. We investigated the effects of the neonicotinoid insecticide thiamethoxam on the homing performances of foragers emerging from colonies differentiated by health status (infestation with Varroa destructor mites, microsporidian parasite Nosema spp. and Deformed Wing Virus). We designed a homing test that has been recently identified to fill a regulatory gap in the field evaluations of sublethal doses of pesticides before their registration. We also assessed the effect of temperature as an environmental factor. Our results showed that the Varroa mite exacerbates homing failure (HF) caused by the insecticide, whereas high temperatures reduce insecticide-induced HF. Through an analytical Effective Dose (ED) approach, predictive modeling results showed that, for instance, ED level of an uninfested colony, can be divided by 3.3 when the colony is infested by 5 Varroa mites per 100 bees and at a temperature of 24 °C. Our results suggest that the health status of honeybee colonies and climatic context should be targeted for a thorough risk assessment.

Beekeeper stewardship, colony loss, and Varroa destructor management.

Varroa (Varroa destructor) is a leading cause of honey bee mortality worldwide. In a U.S. national survey of beekeepers, 3519 respondents noted what they believe are the advantages and disadvantages of managing for Varroa, what good stewardship means in beekeeping, and whether they treated for Varroa. Dominant attitudes were keeping bees healthy, minimizing disturbance, and monitoring hives. We found a bifurcation in Varroa management beliefs. Decision tree analyses show group distinctions. Treatment Skeptics tend to say that stewardship means bees should not be disturbed or subjected to chemicals, and should be given forage to do their 'normal business.' This group was less likely to treat for Varroa. Treatment Adherents identify themselves as bee stewards and say stewardship means active hive management and keeping bees healthy and alive. Illuminating beekeeper stewardship is essential for a socioecological understanding of how to address challenging Varroa management and complex human-environmental production systems that have landscape-level effects.