The requirements of diagnostic methods for the detection of parasites/pathogens are dependent on the overall aim of a study. Epidemiological studies tend to focus on presence/absence studies in order to unravel factors contributing to the spread of disease agents (Kleeman et al., 2002; Njiru et al., 2008; Erler et al., 2012). In contrast, studies driven by evolutionary questions might be interested in the parasite genotypes contributing to infections, in order to determine genotype by genotype interactions, local adaptation, and changes in genotype frequencies predicted by models like the Red Queen or the gene-for-gene theory for host-parasite co-evolution, the last being used mainly for plants (Flor, 1942; Bell, 1982; Decaestecker et al., 2007). Pathology studies, however, in different contexts (ecology and evolution) need the quantification of the infection rate of a host as the primary data. Due to technical advances, molecular diagnostic methods are increasingly used because of their cost effectiveness and reliability (Kelley et al., 2006; Leisova et al., 2006; Leblanc-Maridor et al., 2011). Microscopic methods are popular because very little advanced equipment is needed. However, when dealing with large samples [as in ecological studies (Salath e & Schmid-Hempel, 2011; Goulson et al., 2012)] microscopic methods require a long time to process each sample and to measure the infection rates, and this might result in increasing error rates over long periods of analyses (Pattyn et al., 2008; Lim et al., 2010). Molecular methods are more easily standardized, allowing for integrative studies andmeta-analyses. During the past years, declines of pollinators have been repeatedly reported (Biesmeijer et al., 2006; Ratnieks & Carreck, 2010; Cameron et al., 2011). Factors contributing to pollinator declines have been identified as habitat fragmentation (Murray et al., 2009; Winfree et al., 2009; Potts et al., 2010), pesticide usage (Gill et al., 2012; Henry et al., 2012; Stokstad, 2012;Whitehorn et al., 2012), and increasing parasite loads (Cox-Foster et al., 2007; vanEngelsdorp et al., 2009; Cameron et al., 2011). Thus, studies of parasites in insect pollinators are highly demanded, both from an epidemiological as well as an evolutionary point of view. Amajormodel system in evolutionary ecology of pollinators is the bumblebee (Bombus spec.) (Jordano, 1987). Bumblebees are important pollinators for crops and wild plants (Corbet et al., 1991; Lye et al., 2011; Graystock et al., 2013). Many of their biological aspects have been studied [sociality (Bourke & Ratnieks, 1999; LopezVaamonde et al., 2004; Huth-Schwarz et al., 2011), cognition (Skorupski & Chittka, 2006; Riveros & Gronenberg, 2009), ecology (Kremen et al., 2007), and host-parasite co-evolution (Schmid-Hempel, 1998)]. Bumblebees are eusocial insects with an annual life-cycle, whose colonies are founded by a single-mated queen. Their social life and low genetic diversity within a colony make them a prime target for parasites. Their social organization provides parasites with both a stable and rich environment (Schmid-Hempel, 1998). The low genetic variability within a colony, due to the single-mated and unique queen, allows parasites to easily infect an entire colony (Baer & SchmidHempel, 1999, 2001). One of the most widespread parasites in bumblebees is Crithidia bombi (Protozoa: Trypanosomatidae), a trypanosome gut parasite. Crithidia bombi decreases the chance of successful colony foundation *Correspondence: Bertrand Fouks, Institut fur Biologie,Molekulare Okologie,Martin-Luther-Universitat Halle-Wittenberg, HoherWeg 4, Halle (Saale) 06099, Germany. E-mail: bertrand.fouks@zoologie. uni-halle.de