SummaryHoney bee virus research is an enormously broad area, ranging from subcellular molecular biology through physiology and behaviour, to individual and colony-level symptoms, transmission and epidemiology. The research methods used in virology are therefore equally diverse.This article covers those methods that are very particular to virological research in bees, with numerous cross-referrals to other BEEBOOK papers on more general methods, used in virology as well as other research. At the root of these methods is the realization that viruses at their most primary level inhabit a molecular, subcellular world, which they manipulate and interact with, to produce all higher order phenomena associated with virus infection and disease. Secondly, that viruses operate in an exponential world, while the host operates in a linear world and that much of the understanding and management of viruses hinges on reconciling these fundamental mathematical differences between virus and host. The article concentrates heavily on virus propagation and methods for detection, with minor excursions into surveying, sampling management and background information on the many viruses found in bees. Métodos estándar para la investigación de virus en Apis mellifera ResumenLa investigación de los virus de la abeja de la miel es un área sumamente amplia, que abarca desde la biología molecular subcelular hasta la fisiología y el comportamiento, desde síntomas al nivel de individuo hasta al nivel de la colmena, transmisión y epidemiología. Los métodos de investigación en virología son, por tanto, diversos. Este artículo incluye aquellos métodos específicos de la investigación virológica en las abejas, con numerosas referencias cruzadas con otros artículos del BEEBOOK y otros más generales, usados tanto en virología como en otras disciplinas. La base de estos métodos es la comprensión de los virus en su nivel primario de hábitat molecular, ambiente subcelular, que manipulan y con el que interactúan, para producir otros fenómenos de orden superior asociados a la infección del virus y la enfermedad. En segundo lugar, estos virus actúan en un mundo exponencial, mientras que los hospedadores actúan en un mundo lineal y gran parte del entendimiento y manejo de los virus depende de los fundamentos matemáticos de las diferencias entre el virus y el hospedador. El artículo se centra principalmente en la propagación de virus y en los métodos para su detección, con inclusiones menores en su estudio, el manejo del muestreo y la información general sobre los numerosos virus que se encuentran en las abejas.

No abstract

SummaryA variety of methods are used in honey bee research and differ depending on the level at which the research is conducted. On an individual level, the handling of individual honey bees, including the queen, larvae and pupae are required. There are different methods for the immobilising, killing and storing as well as determining individual weight of bees. The precise timing of developmental stages is also an important aspect of sampling individuals for experiments. In order to investigate and manipulate functional processes in honey bees, e.g. memory formation and retrieval and gene expression, microinjection is often used. A method that is used by both researchers and beekeepers is the marking of queens that serves not only to help to locate her during her life, but also enables the dating of queens. Creating multiple queen colonies allows the beekeeper to maintain spare queens, increase brood production or ask questions related to reproduction. On colony level, very useful techniques are the measurement of intra hive mortality using dead bee traps, weighing of full hives, collecting pollen and nectar, and digital monitoring of brood development via location recognition. At the population level, estimation of population density is essential to evaluate the health status and using beelines help to locate wild colonies. These methods, described in this paper, are especially valuable when investigating the effects of pesticide applications, environmental pollution and diseases on colony survival. Métodos estándar diversos para la investigación en Apis mellifera ResumenEn la investigación de la abeja de la miel, se han usado una variedad de métodos que se diferencian en función del nivel en el que se realiza la investigación. Al nivel individual, el manejo de las abejas individuales es necesario, incluyendo a la reina, las larvas y las pupas. Existen diferentes métodos para la inmovilización, mortandad y almacenamiento, así como para la determinación del peso individual de las abejas. La precisión en la sincronización de las etapas de desarrollo es también un aspecto importante de los experimentos con muestreos individuales.La microinyección se utiliza a menudo con el fin de investigar y manipular los procesos funcionales de las abejas melíferas, como por ejemplo, la formación y recuperación de la memoria y la expresión génica. Un método utilizado tanto por investigadores como apicultores es el marcado de las reinas, que sirve no sólo para ayudar a localizarlas durante su vida, sino que también permite su datación. La creación de varias colmenas a partir de reinas permite al apicultor mantener reinas de repuesto, aumentar la producción de cría o hacer preguntas relacionadas con la reproducción. Al nivel de colmena, la medición de la mortalidad intra colmena utilizando trampas de abejas muertas, el pesaje de las colmenas completas, la recolección de polen y néctar, y el seguimiento digital del desarrollo de la cría a través del reconocimiento de su ubicación, son algunas de las técnicas más útiles. Al nivel p...

BackgroundRecent elevated winter loss of honey bee colonies is a major concern. The presence of the mite Varroa destructor in colonies places an important pressure on bee health. V. destructor shortens the lifespan of individual bees, while long lifespan during winter is a primary requirement to survive until the next spring. We investigated in two subsequent years the effects of different levels of V. destructor infestation during the transition from short-lived summer bees to long-lived winter bees on the lifespan of individual bees and the survival of bee colonies during winter. Colonies treated earlier in the season to reduce V. destructor infestation during the development of winter bees were expected to have longer bee lifespan and higher colony survival after winter.Methodology/Principal FindingsMite infestation was reduced using acaricide treatments during different months (July, August, September, or not treated). We found that the number of capped brood cells decreased drastically between August and November, while at the same time, the lifespan of the bees (marked cohorts) increased indicating the transition to winter bees. Low V. destructor infestation levels before and during the transition to winter bees resulted in an increase in lifespan of bees and higher colony survival compared to colonies that were not treated and that had higher infestation levels. A variety of stress-related factors could have contributed to the variation in longevity and winter survival that we found between years.Conclusions/SignificanceThis study contributes to theory about the multiple causes for the recent elevated colony losses in honey bees. Our study shows the correlation between long lifespan of winter bees and colony loss in spring. Moreover, we show that colonies treated earlier in the season had reduced V. destructor infestation during the development of winter bees resulting in longer bee lifespan and higher colony survival after winter.

SummaryModern agriculture often involves the use of pesticides to protect crops. These substances are harmful to target organisms (pests and pathogens). Nevertheless, they can also damage non-target animals, such as pollinators and entomophagous arthropods. It is obvious that the undesirable side effects of pesticides on the environment should be reduced to a minimum. Western honey bees (Apis mellifera) are very important organisms from an agricultural perspective and are vulnerable to pesticide-induced impacts. They contribute actively to the pollination of cultivated crops and wild vegetation, making food production possible. Of course, since Apis mellifera occupies the same ecological niche as many other species of pollinators, the loss of honey bees caused by environmental pollutants suggests that other insects may experience a similar outcome. Because pesticides can harm honey bees and other pollinators, it is important to register pesticides that are as selective as possible. In this manuscript, we describe a selection of methods used for studying pesticide toxicity/selectiveness towards Apis mellifera. These methods may be used in risk assessment schemes and in scientific research aimed to explain acute and chronic effects of any target compound on Apis mellifera. Métodos estándar para la investigación toxicológica en Apis mellifera ResumenLa agricultura moderna a menudo implica el uso de plaguicidas para proteger los cultivos. Estas sustancias son dañinas para los organismos objetivo (plagas y patógenos). Sin embargo, también pueden dañar a animales que no son objetivo, como artrópodos polinizadores y entomófagos. Obviamente los efectos secundarios indeseables de los plaguicidas sobre el medio ambiente deben ser reducidos al mínimo. Las abejas occidentales (Apis mellifera) son organismos muy importantes desde el punto de vista agrícola y son vulnerables a los impactos inducidos por los plaguicidas. Contribuyen activamente a la polinización de los cultivos y de la vegetación silvestre, lo que hace posible la producción de alimentos. Como Apis mellifera ocupa el mismo nicho ecológico que muchas otras especies de polinizadores, la pérdida de las abejas melíferas causada por contaminantes ambientales sugiere que otros insectos pueden experimentar un resultado similar. Ya que los plaguicidas pueden dañar a las abejas y a otros polinizadores, es importante registrar los plaguicidas que sean lo más selectivos posible. En este artículo, se describe una selección de los métodos utilizados para el estudio de la toxicidad y el efecto selectivo de los plaguicidas hacia Apis mellifera. Estos métodos se pueden utilizar en sistemas de evaluación de riesgo y en la investigación científica para explicar los efectos agudos y crónicos en Apis mellifera de cualquier compuesto objetivo.

No abstract

SummaryIn this article we provide guidelines on statistical design and analysis of data for all kinds of honey bee research. Guidelines and selection of different methods presented are, at least partly, based on experience. This article can be used: to identify the most suitable analysis for the type of data collected; to optimise one's experimental design based on the experimental factors to be investigated, samples to be analysed, and the type of data produced; to determine how, where, and when to sample bees from colonies; or just to inspire. Also included are guidelines on presentation and reporting of data, as well as where to find help and which types of software could be useful. Guia estadistica para estudios en Apis mellifera ResumenEn este trabajo se proporcionan directrices sobre el diseño estadístico y el análisis de datos para todo tipo de investigación sobre abejas.Tanto las directrices como la selección de los diferentes métodos que se presentan están basadas, al menos en parte, en la experiencia. Este artículo se puede utilizar: para identificar el análisis más adecuado para el tipo de datos recogidos; para optimizar el diseño experimental basado en los factores experimentales a ser investigados, las muestras a analizar, y el tipo de datos que se producen; para determinar cómo, dónde , y cuando muestras abejas de las colonias, o simplemente para inspirar. También se incluyen directrices para la presentación y comunicación de los datos, así como dónde encontrar ayuda y distintos software que puedan ser útiles.

Honeybees (Apis mellifera L.) have great potential for detecting and monitoring environmental pollution, given their wide-ranging foraging behaviour. Previous studies have demonstrated that concentrations of metals in adult honeybees were significantly higher at polluted than at control locations. These studies focused at a limited range of heavy metals and highly contrasting locations, and sampling was rarely repeated over a prolonged period. In our study, the potential of honeybees to detect and monitor metal pollution was further explored by measuring the concentration in adult honeybees of a wide range of trace metals, nine of which were not studied before, at three locations in the Netherlands over a 3-month period. The specific objective of the study was to assess the spatial and temporal variation in concentration in adult honeybees of Al, As, Cd, Co, Cr, Cu, Li, Mn, Mo, Ni, Pb, Sb, Se, Sn, Sr, Ti, V and Zn. In the period of July–September 2006, replicated samples were taken at 2-week intervals from commercial-type beehives. The metal concentration in micrograms per gram honeybee was determined by inductive coupled plasma–atomic emission spectrometry. Significant differences in concentration between sampling dates per location were found for Al, Cd, Co, Cr, Cu, Mn Sr, Ti and V, and significant differences in average concentration between locations were found for Co, Sr and V. The results indicate that honeybees can serve to detect temporal and spatial patterns in environmental metal concentrations, even at relatively low levels of pollution.