Bio-sampling is a function of bio-indication. Bio-indication with honeybee colonies (Apis mellifera L) is where the research fields of environmental technology and apiculture overlap. The honeybees are samplers of the environment by collecting unintentionally and simultaneously, along with nectar, pollen, water and honeydew from the flowers or on the leaves, other matter (in bio-indication terms: target matter) and accumulating this in the colony. Collected target matter, in this thesis heavy metals, the plant pathogens Erwinia pyrifoliae and Erwinia amylovora and the soil pollutant γ-HCH, is collected from the colony by subsampling. Subsampling the honeybee colony is done by taking and killing bees from the hive (sacrificial) or by collecting target matter from the bee’s exterior without killing the bee (non-sacrificial). In environmental technology terms the application of the honeybee colony is a Passive Sampling Method (PSM). In this thesis the possibilities and restrictions of the PSM honeybee colony are explored. Bio-indication is a broad research field with one common factor: a living organism (bio) is applied to record an alteration of the environment (indication). The environment may be small such as a laboratory or big such as an ecosystem. Alterations in the organism may vary from detecting substances foreign to the body to mortality of the organism. In environmental technology the concept Source-Path-Receptor (SPR) is applied to map the route of a pollutant. It describes where in the environment the pollution is, how it moves through the environment and where it ends. This environment is the same environment of all living organisms, ergo also honeybees. Honeybees depend on flowers for their food. In the SPR concept, a flower can be a source, path or receptor. Along with collecting pollen, nectar, water and honeydew, target matter is collected by honeybees. Each honeybee functions as a micro-sampler of target matter in the environment, in this case the flower. Each honeybee is part of a honeybee colony and in fact the honeybee colony is the bio-sampler. The honeybee colony is a superorganism. The well-being of the colony prevails over the individual honeybee. Food collection is directed by the colony’s need. Foragers are directed to the most profitable food sources by the bee dance and food exchange (trophallaxis). The result of this feature is that mainly profitable sources are exploited and poor food sources less or not at all. During the active foraging period hundreds to thousands of flowers are visited daily. The nectar, pollen, water and honeydew plus the unintentionally collected target matter is accumulated in the honeybee colony. In order to obtain target matter the colony must be subsampled. This is done by picking bees from the hive-entrance (hive-entering bees) or inside the hive (in-hive bees) and processing them for analysis (sacrificial). This is the most commonly applied method. However, it is possible to subsample the colony without picking and processing the bees by collecting target matter from the hive-entering bee’s exterior (non-sacrificial). For non-sacrificial subsampling of the honeybee colony the Beehold device with the sampling part Beehold tube has been developed. The results of bio-indication with honeybee colonies are qualitative and indicative for follow up study (Chapter 1). Six bio-indication studies with honeybee colonies for bio-indication of heavy metals, the plant pathogens Erwinia pyrifoliae and Erwinia amylovora and the soil pollutant γ-HCH are presented. Chapter 2 describes how the concentration of eighteen heavy metals in honeybees fluctuate throughout the period of July, August and September (temporal) at the study sites: the city of Maastricht, the urban location with an electricity power plant in Buggenum and along the Nieuwe Waterweg at Hoek van Holland (spatial). A number of the metals have not been previously analysed in honeybees. To study whether honeybees can be used for bio-indication of air pollution, the concentrations of cadmium, vanadium and lead were compared to concentrations found in honeybees. The honeybee colonies were placed next to the air samplers. Only significant differences of metal concentrations in the ambient air also show in honeybees. This was the case with vanadium in ambient air and honeybees. The spatial and temporal differences of cadmium and lead were too futile to demonstrate a correspondence (Chapter 3). In a national surveillance study in 2008 the concentration of eighteen metals in honeybees has been analysed. The results showed a distinct regional pattern. Honeybees in the East of the Netherlands have higher concentrations of heavy metals compared to the bees in the West. Besides regional differences local differences were also recorded. An approximate description of the land use around 148 apiaries (> 50% agriculture, > 50% wooded area, > 50% urban area and mixed use) indicated the impact of land use on metal concentrations in honeybees. In areas with > 50% wood significantly higher concentrations of heavy metals were detected (Chapter 4). Subsampling of the honeybee colonies in Chapter 2, 3 and 4 was done sacrificially. In the studies presented in Chapter 5, 6, and 7 the honeybee colonies were subsampled non-sacrificially or simultaneously non-sacrificially and sacrificially. The plant pathogen E. pyrifoliae causes a flower infection in the strawberry cultivation in greenhouses. In greenhouse strawberry cultivation honeybees are applied for pollination. In Chapter 5 the combination pollination / bio-indication by honeybee colonies is studied. This proved to be a match. E. pyrifoliae could be detected on in-hive bees prior to any symptom of the infection in the flowers. In the Beehold tube, the bacterium was detected at the same time as the first tiny symptoms of the infection. In Chapter 5 the principles on which the Beehold tube is based are presented and discussed. The plant pathogen E. amylovora causes fireblight in orchards. The combination pollination / bio-indication has also been applied in this study performed in Austria in 2013. It is known that E. amylovora can be detected on honeybees prior to any symptom in the flower or on the fruit tree. A fireblight outbreak depends on flowering period, humidity and temperature. In 2013 no fireblight infection emerged in the orchards where the study was performed. Therefore, the bacterium could not be detected on the honeybees. γ-HCH (Lindane) is one of the soil pollutants in the Bitterfeld region in Saxony-Anhalt in Germany. It is the result of dumping industrial waste around the production locations. Although γ-HCH is bound to soil particles there is a flux to groundwater and surface water. Consequently, the pollution may end up in the sediments of the streambed and flood plains. The study objective was to investigate the hypothetic route of γ-HCH from polluted soil (source), via soil erosion and atmospheric deposition (route) to the receptor (flowering flowers) by detecting γ-HCH in the Beehold tube. Although on average over 17000 honeybees passed through the Beehold tube daily for a maximal period of 28 days, no γ-HCH has been detected. The pollen pattern in the Beehold tube revealed where the bees collected the food (Chapter 7). The application of the honeybee colony has pros and cons. Distinctive pros are many micro samplers, the extensive collection of matter (both food and target matter) and the accumulation in the colony. For successful bio-indication with honeybee colonies, determining factors are: the target matter, location of the target matter, distance between target matter and the honeybee colony, individual or pooled subsampling, the minimal sampling frequency and sample size, and sacrificial or non-sacrificial subsampling applied solely or in combination. Taking bees from a colony impacts upon the colony’s performance and consequently the passive sampling method. Based on a long-years’ experience and inter-collegial discussion it is stated that 3% of the forager bees (hive-entering) and 1.5% of the in-hive bees can be sampled safely without impacting upon the colony. This restriction does not apply when carrying out non-sacrificial subsampling of the honeybee colony (Chapter 8). Performing bio-indication with honeybee colonies has more applications than have been exploited so far. Further research can make a change. In particular I mention here the combination of pollination and bio-indication and the application of non-sacrificial subsampling solely or in combination with sacrificial subsampling. Everywhere Apiculture is practiced (all over the world except the polar areas) bio-indication with honeybee colonies can be applied in a simple, practical and low cost way.