Search Results (11)

The increasing reliance of modern agriculture on honey bee (Apis mellifera) pollination has driven efforts to preserve and enhance bee populations. The cryopreservation of drone semen presents a promising solution for preserving genetic diversity and supporting breeding programs without live animal transport risks. This study aimed to evaluate a one-step dilution antibiotic-free drone semen slow-freezing protocol under field conditions with in vitro and in vivo parameters. Semen viability was tested by two different mixes of dyes, and both techniques gave similar results, showing a post-thaw viability drop of 37%. Virgin queens were inseminated either with fresh or frozen–thawed semen. Survival rates until egg-laying onset and female brood production were similar for both groups; however, colonies with queens inseminated with fresh semen were more likely to go into wintering. Results suggest that frozen–thawed semen can support viable queen insemination, with potential for female brood production even without antibiotics in the diluent. This study highlights the need for further refinement of cryopreservation protocols, particularly regarding semen quality and queen longevity, to improve the feasibility of cryobanking for Apis mellifera conservation and breeding. Full article

The study aimed to verify whether urban beekeeping affects the strength of the honeybee (Apis mellifera) colonies from urban apiaries and the variability of the crucial for their health and long-life protein—vitellogenins. For this purpose, honeybees were kept in two locations—in a city apiary on a roof in the city center and in agricultural areas. Each of the apiaries consisted of six colonies, with the sister queens artificially inseminated with semen from the same pool of drones. The bee colony strength and the variability of the vitellogenins in various tissues in foragers from both apiaries were analyzed from May to August. Here, we revealed that colonies from the urban apiary were more abundant than those from the rural apiary. We observed the compensation mechanism during periods of worker deficiency in the bee colony, which was expressed as a change in the Vgs level in the forager tissues. Using the vitellogenin level as a biomarker of the honeybee colony strength can predict the fate of colonies, especially those with low numbers. The high level of Vgs can be a candidate for bee colony depopulation biomarker. Full article

Artificial insemination in queen honey bees is the only tool that provides complete control over mating for research and breeding purposes, making it essential in genetic improvement and conservation programs in this species. The aims of this study were to characterize drone semen bacterial loads by culture-dependent and independent methods and to describe their variation depending on the method of semen collection, the colony and the apiary. In the first experiment, the bacterial loads of semen collected from the seminal vesicles or from ejaculates was studied using culture-dependent methods. The collection method had a significant influence on the overall bacterial count in semen. Out of the 42 semen samples analyzed, 26 (61.9%) tested positive for bacterial isolation. This encompassed the entirety of samples obtained from the seminal vesicles (21 of 21), whereas only 23.8% of those derived from ejaculates (5 out of 21) showed bacterial isolation. In the second experiment, next-generation sequencing techniques were used to describe the microbiome of ejaculated drone semen for the first time. The most abundant phyla were Proteobacteria, Firmicutes, Bacteroidota and Actinobacteriota, while the most abundant genera were Lactobacillus, Staphylococcus, Prevotella, Alloprevotella and Streptococcus. The results showed that the apiary had a significant effect on the community structure composition and abundance of the seminal microbiota, and significative differences in abundance were observed for the genera Sphingomonas, Methylobacterium-Methylorubrum, Bifidobacterium and Alloprevotella. Significant differences were also observed in the richness of the microbiota between apiaries and colonies. Full article

Many fluorochromes routinely used in semen quality analysis emit in the green and red channels, limiting their possible combination for multiple parameter analysis. The use of fluorophores emitting in different light channels broadens the possibilities of combination to expand the range of simultaneously evaluated criteria. This is of great interest in cases of small ejaculated volumes, such as those naturally occurring in roosters, small dog breeds and drones (Apis mellifera). The purpose of this experiment is to establish Calcein Violet (CaV), a blue fluorochrome, as a marker of viability and acrosomal integrity in domestic animals in order to free the red and green channels. SYBR^®14/Propidium Iodide (PI) was used as reference dye, heat-treated samples as negative controls, serial staining combination for validation and epifluorescence microscopy for observation. Dead spermatozoa marked in red with PI showed no blue fluorescence either from the head or the tail. Live spermatozoa showed a decreasing blue emission from head to tail when single stained with CaV. Unreacted acrosomes showed intense blue fluorescence irrespective of plasma membrane integrity. This needs to be further confirmed for species with small and difficult to observe heads. Establishment of CaV as a marker of membrane integrity by fluorescence microscopy is a decisive first step towards further technical development and use with flow cytometry. Full article

The quantity and quality of the honey bee drone semen have a significant determination on the performance of bee colonies. The existence of a smaller number of mature drones to participate in the mating of queens, as well as a sufficient number of drones but with poor quality semen can have serious implications for the productivity of bee colonies. Our study aimed to investigate the correlation between two body weight ranges of drones and semen parameters in the Buckfast honey bee, data that could be integrated into the optimization of instrumental insemination in been queens. Semen was collected from two groups of drones with different body weights (200–240 mg and 240–280 mg). Semen volume, semen concentration, motility, morphology and membrane integrity of spermatozoa were analyzed. The phenotype indicator related to body weight in correlation with the main semen parameters studied gives a weak influence or causality ratio. In drones with 240–280 mg body weight, a higher percentage of spermatozoa with abnormal morphology (>9.60%) was recorded, compared to drones with 200–240 mg body weight. The study reveals that a higher weight of honey bee drones is correlated with higher sperm concentration and total number of spermatozoa/ejaculate, with an increase in the percentage of spermatozoa with abnormal morphology. Full article

Plant protection products (PPPs) are pesticides that protect crops and ornamental plants. PPPs include primarily insecticides, herbicides, and fungicides. Bees’ contact with PPPs can cause immediate death or, in sublethal dose, may affect their physiology and/or behavior. Understanding the effect of PPPs’ sublethal doses is especially important. Contact with a sublethal dose of PPPs generally allows the bee to return to the hive, which may expose the whole colony to the harmful substance. Biochemical changes may affect colony condition, health, and performance. Most of the research on the biochemical effects of PPP in honey bees focuses on insecticides and among them neonicotinoids (especially imidacloprid). The vast majority of research is carried out on Apis mellifera workers. A small part of the research has been conducted on drones and queens. Pesticides, including fungicides and herbicides, may alter antioxidant defense, detoxification, gene expression, and immune response of the bee. They affect the drones’ semen quality and metabolic rate of the queen. In this review, the biochemical effect of PPP products in the honey bee was examined, with a focus on the effect on cytochrome P450 monooxygenases, glutathione transferases, and carboxylesterases, which take part in toxin metabolism or the detoxification process. PPPs effects on the activity of glutathione peroxidase (GPX), catalase (CAT), superoxide dismutase (SOD), proteases, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and phenoloxidase (PO) are also presented. Full article

Bumblebees and honeybees are very important pollinators and play a vital role in agricultural and natural ecosystems. The quality of their colonies is determined by the queens and the reproductive drones of mother colonies, and mated drones transmit semen, including half of the genetic materials, to queens and enhance their fertility. Therefore, factors affecting drone fecundity will also directly affect progeny at the colony level. Here, we review environmental and bee-related factors that are closely related to drone reproductive ability. The environmental factors that mainly affect the sperm count and the viability of males include temperature, nutrients and pesticides. In addition, the inherent characteristics of male bees, such as body size, weight, age, seminal fluid proteins and proteins of the spermathecal fluid, contribute to mating success, sperm quality during long-term storage in the spermathecae and the reproductive behaviors of queens. Based on the results of previous studies, we also suggest that the effects of somatotype dimorphism in bumblebee males on sperm quality and queen fecundity and the indispensable and exploitable function of gland proteins in the fecundity of males and queens should be given more attention in further studies. Full article

The quality of honey bee drone semen is relevant in different contexts, ranging from colony productivity to pathology, toxicology and biodiversity preservation. Despite its importance, considerably less knowledge is available on this subject for the honey bee when compared to other domestic animal species. A proper assessment of sperm quality requires a multiple testing approach which discriminates between the different aspects of sperm integrity and functionality. Most studies on drone semen quality have only assessed a few parameters, such as sperm volume, sperm concentration and/or sperm plasma membrane integrity. Although more recent studies have focused on a broader variety of aspects of semen quality, some techniques currently used in vertebrates, such as computer-assisted sperm analysis (CASA) or multiparametric sperm quality testing, still remain to be developed in the honey bee. This may be attributed to the particular sperm morphology and physiology in this species, requiring the development of technologies specifically adapted to it. This article reviews the present knowledge of sperm quality in honey bee drones, highlighting its peculiarities and proposing future lines of research. Full article

Instrumental insemination of Apis mellifera L. queens is a widely employed technique used in honeybee breeding that enables the effective control of mating. However, drone semen represents a potential source of honeybee viruses. In this study, 43 semen doses collected from apparently healthy drones, and consequently used in instrumental insemination, were analysed using PCR or RT-PCR to detect the presence of viral genome of 11 honeybee viruses. In 91% of samples, viral infection was detected. The survey revealed genomes of five viruses, namely Deformed wing virus (DWV), Acute bee paralysis virus (ABPV), Black queen cell virus (BQCV), Sacbrood virus (SBV), and A. mellifera filamentous virus (AmFV) in 84%, 19%, 14%, 2%, and 67% of samples, respectively. Single infection (30% of samples) as well as multiple infection (61% of samples) of two, three or four pathogens were also evaluated. To the best of our knowledge, this is the first study describing the presence of the BQCV and SBV genome sequence in drone ejaculate. Phylogenetic analysis of BQCV partial helicase gene sequence revealed the high similarity of nucleotide sequence of described Czech strains, which varied from 91.4% to 99.6%. The findings of our study indicate the possibility of venereal transmission of BQCV and SBV. Full article

In the face of high proportions of yearly colony losses, queen health and fecundity has been a major focus of industry and research. Much of the reproductive quality of the queen, though, is a function of the mating success and quality of the drones (males). Many environmental factors can negatively impact drone semen quality, but little is known about factors that impact the drones’ ability to successfully mate and deliver that semen, or how widely drones vary. In our study, we observed the daily variation in honey bee drone reproductive quality over time, along with a number of morphological traits. Drones were reared in cages in bank colonies, and 20 individuals were dissected and measured daily. The number of viable spermatozoa in the seminal vesicles was zero at emergence and reached an average maximum of 7.39 ± 0.19 million around 20 days of life. Decline in spermatozoa count occurred after day 30, though viability was constant throughout life, when controlling for count. Older drones had smaller wet weights, head widths, and wing lengths. We predict that this is likely due to sampling bias due to a differential lifespan among larger, more reproductively developed drones. Our study shows that drones are more highly variable than previously suggested and that they have a significant variation in reproductive physiology as a function of age. Full article

Honey bees are major pollinators of agricultural and non-agricultural landscapes. In recent years, honey bee colonies have exhibited high annual losses and commercial beekeepers frequently report poor queen quality and queen failure as the primary causes. Honey bee colonies are highly vulnerable to compromised queen fertility, as each hive is headed by one reproductive queen. Queens mate with multiple drones (male bees) during a single mating period early in life in which they obtain enough spermatozoa to fertilize their eggs for the rest of their reproductive life span. The process of mating initiates numerous behavioral, physiological, and molecular changes that shape the fertility of the queen and her influence on the colony. For example, receipt of drone semen can modulate queen ovary activation, pheromone production, and subsequent worker retinue behavior. In addition, seminal fluid is a major component of semen that is primarily derived from drone accessory glands. It also contains a complex mixture of proteins such as proteases, antioxidants, and antimicrobial proteins. Seminal fluid proteins are essential for inducing post-mating changes in other insects such as Drosophila and thus they may also impact honey bee queen fertility and health. However, the specific molecules in semen and seminal fluid that initiate post-mating changes in queens are still unidentified. Herein, we summarize the mating biology of honey bees, the changes queens undergo during and after copulation, and the role of drone semen and seminal fluid in post-mating changes in queens. We then review the effects of seminal fluid proteins in insect reproduction and potential roles for honey bee drone seminal fluid proteins in queen reproduction and health. We finish by proposing future avenues of research. Further elucidating the role of drone fertility in queen reproductive health may contribute towards reducing colony losses and advancing honey bee stock development. Full article