David R. Tarpy

Colony Collapse Disorder: A Descriptive Study

Background Over the last two winters, there have been large-scale, unexplained losses of managed honey bee (Apis mellifera L.) colonies in the United States. In the absence of a known cause, this syndrome was named Colony Collapse Disorder (CCD) because the main trait was a rapid loss of adult worker bees. We initiated a descriptive epizootiological study in order to better characterize CCD and compare risk factor exposure between populations afflicted by and not afflicted by CCD. Methods and Principal Findings Of 61 quantified variables (including adult bee physiology, pathogen loads, and pesticide levels), no single measure emerged as a most-likely cause of CCD. Bees in CCD colonies had higher pathogen loads and were co-infected with a greater number of pathogens than control populations, suggesting either an increased exposure to pathogens or a reduced resistance of bees toward pathogens. Levels of the synthetic acaricide coumaphos (used by beekeepers to control the parasitic mite Varroa destructor) were higher in control colonies than CCD-affected colonies. Conclusions/Significance This is the first comprehensive survey of CCD-affected bee populations that suggests CCD involves an interaction between pathogens and other stress factors. We present evidence that this condition is contagious or the result of exposure to a common risk factor. Potentially important areas for future hypothesis-driven research, including the possible legacy effect of mite parasitism and the role of honey bee resistance to pesticides, are highlighted.

Genetic diversity within honeybee colonies prevents severe infections and promotes colony growth.

Multiple mating by social insect queens increases the genetic diversity among colony members, thereby reducing intracolony relatedness and lowering the potential inclusive fitness gains of altruistic workers. Increased genetic diversity may be adaptive, however, by reducing the prevalence of disease within a nest. Honeybees, whose queens have the highest levels of multiple mating among social insects, were investigated to determine whether genetic variation helps to prevent chronic infections. I instrumentally inseminated honeybee queens with semen that was either genetically similar (from one male) or genetically diverse (from multiple males), and then inoculated their colonies with spores of Ascosphaera apis, a fungal pathogen that kills developing brood. I show that genetically diverse colonies had a lower variance in disease prevalence than genetically similar colonies, which suggests that genetic diversity may benefit colonies by preventing severe infections.

Pathogen Webs in Collapsing Honey Bee Colonies

Recent losses in honey bee colonies are unusual in their severity, geographical distribution, and, in some cases, failure to present recognized characteristics of known disease. Domesticated honey bees face numerous pests and pathogens, tempting hypotheses that colony collapses arise from exposure to new or resurgent pathogens. Here we explore the incidence and abundance of currently known honey bee pathogens in colonies suffering from Colony Collapse Disorder (CCD), otherwise weak colonies, and strong colonies from across the United States. Although pathogen identities differed between the eastern and western United States, there was a greater incidence and abundance of pathogens in CCD colonies. Pathogen loads were highly covariant in CCD but not control hives, suggesting that CCD colonies rapidly become susceptible to a diverse set of pathogens, or that co-infections can act synergistically to produce the rapid depletion of workers that characterizes the disorder. We also tested workers from a CCD-free apiary to confirm that significant positive correlations among pathogen loads can develop at the level of individual bees and not merely as a secondary effect of CCD. This observation and other recent data highlight pathogen interactions as important components of bee disease. Finally, we used deep RNA sequencing to further characterize microbial diversity in CCD and non-CCD hives. We identified novel strains of the recently described Lake Sinai viruses (LSV) and found evidence of a shift in gut bacterial composition that may be a biomarker of CCD. The results are discussed with respect to host-parasite interactions and other environmental stressors of honey bees.

Queen promiscuity lowers disease within honeybee colonies

Most species of social insects have singly mated queens, but in some species each queen mates with numerous males to create a colony with a genetically diverse worker force. The adaptive significance of polyandry by social insect queens remains an evolutionary puzzle. Using the honeybee (Apis mellifera), we tested the hypothesis that polyandry improves a colonys resistance to disease. We established colonies headed by queens that had been artificially inseminated by either one or 10 drones. Later, we inoculated these colonies with spores of Paenibacillus larvae, the bacterium that causes a highly virulent disease of honeybee larvae (American foulbrood). We found that, on average, colonies headed by multiple-drone inseminated queens had markedly lower disease intensity and higher colony strength at the end of the summer relative to colonies headed by single-drone inseminated queens. These findings support the hypothesis that polyandry by social insect queens is an adaptation to counter disease within their colonies.

Estimating effective paternity number in social insects and the effective number of alleles in a population

Estimating paternity and genetic relatedness is central to many empirical and theoretical studies of social insects. The two important measures of a queens mating number are her actual number of mates and her effective number of mates. Estimating the effective number of mates is mathematically identical to the problem of estimating the effective number of alleles in population genetics, a common measure of genetic variability introduced by Kimura & Crow (1964 ). We derive a new bias‐corrected estimator of effective number of types (mates or alleles) and compare this new method to previous methods for estimating true and effective numbers of types using Monte Carlo simulations. Our simulation results suggest that the examined estimators of the true number of types have very similar statistical properties, whereas the estimators of effective number of types have quite different statistical properties. Moreover, our new proposed estimator of effective number of types is approximately unbiased, and has considerably lower variance than the original estimator. Our new method will help researchers more accurately estimate intracolony genetic relatedness of social insects, which is an important measure in understanding their ecology and social behaviour. It should also be of use in population genetic studies in which the effective number of alleles is of interest.

Lower disease infections in honeybee (Apis mellifera) colonies headed by polyandrous vs monandrous queens

We studied the relationship between genetic diversity and disease susceptibility in honeybee colonies living under natural conditions. To do so, we created colonies in which each queen was artificially inseminated with sperm from either one or ten drones. Of the 20 colonies studied, 80% showed at least one brood disease. We found strong differences between the two types of colonies in the infection intensity of chalkbrood and in the total intensity of all brood diseases (chalkbrood, sacbrood, American foulbrood, and European foulbrood) with both variables lower for the colonies with higher genetic diversity. Our findings demonstrate that disease can be an important factor in the ecology of honeybee colonies and they provide strong support for the disease hypothesis for the evolution of polyandry by social insect queens.

A scientific note on the revised estimates of effective paternity frequency in Apis

SummaryWe review the literature for studies that investigated paternity within honey bee colonies, and we report the average paternity numbers and effective paternity frequencies of eight species of Apis. Moreover, we employ a recently developed estimator of effective paternity, and we demonstrate that it has little effect on the average paternity estimates but that it can have a profound effect on the variance of individual estimates.

Sampling Error, Effective Paternity, and Estimating the Genetic Structure of Honey Bee Colonies (Hymenoptera: Apidae)

Abstract Multiple mating by social Hymenopteran queens significantly lowers the average genetic relatedness among female nestmates, which subsequently affects a wide range of social behaviors. Honey bees (Apis spp.) have among the highest levels of multiple mating in social insects, and have received the most empirical effort to quantify the effective paternities within colonies. We reviewed 24 studies that estimated paternity frequencies of individual, naturally-mated honey bee queens using molecular techniques. We summarize the methods used to estimate effective paternity (me) and intracolonial genetic relatedness (G). We then concentrate on the effect of sample size on estimates of me using Monte Carlo simulations. The results demonstrate that me estimates may vary significantly as a result of sampling error, particularly at low worker sample sizes and high paternity numbers. From these simulations, we arbitrarily define a “threshold” worker sample size to effective paternity ratio (n/me) that, at best, reduces the error of estimating me to less than one subfamily. The literature review illustrates that no study with an n/me ratio above this threshold estimates an me above 15 subfamilies. Finally, we briefly discuss other factors that may serve to over-estimate me, including numerous sampling biases. We conclude that although 152 colonies in the various species of Apis have been tested, the extremity of their paternity frequencies may be somewhat exaggerated, although not drastically.

Weighing Risk Factors Associated With Bee Colony Collapse Disorder by Classification and Regression Tree Analysis

ABSTRACT Colony collapse disorder (CCD), a syndrome whose defining trait is the rapid loss of adult worker honey bees, Apis mellifera L., is thought to be responsible for a minority of the large overwintering losses experienced by U.S. beekeepers since the winter 2006–2007. Using the same data set developed to perform a monofactorial analysis (PloS ONE 4: e6481, 2009), we conducted a classification and regression tree (CART) analysis in an attempt to better understand the relative importance and interrelations among different risk variables in explaining CCD. Fifty-five exploratory variables were used to construct two CART models: one model with and one model without a cost of misclassifying a CCD-diagnosed colony as a non-CCD colony. The resulting model tree that permitted for misclassification had a sensitivity and specificity of 85 and 74%, respectively. Although factors measuring colony stress (e.g., adult bee physiological measures, such as fluctuating asymmetry or mass of head) were important discriminating values, six of the 19 variables having the greatest discriminatory value were pesticide levels in different hive matrices. Notably, coumaphos levels in brood (a miticide commonly used by beekeepers) had the highest discriminatory value and were highest in control (healthy) colonies. Our CART analysis provides evidence that CCD is probably the result of several factors acting in concert, making afflicted colonies more susceptible to disease. This analysis highlights several areas that warrant further attention, including the effect of sublethal pesticide exposure on pathogen prevalence and the role of variability in bee tolerance to pesticides on colony survivorship.

Colony Collapse Disorder in context

Although most of humanity relies upon foods that do not require animal pollination 1, production of 39 of the worlds 57 most important monoculture crops still benefits from this ecosystem service 2. Western honey bees (Apis mellifera) are undoubtedly the single-most valuable animal pollinators to agriculture because they can be easily maintained and transported to pollinator-dependent crops. Yet, despite an almost 50% increase in world honey bee stocks over the last century, beekeepers have not kept pace with the >300% increase in pollinator-dependent crops 3. This has led to great uncertainty surrounding the recent large-scale die-offs of honey bees around the world, and has sparked enormous interest from both scientists and the general public.