Search

Bumblebees are highly valued for their pollination services in natural ecosystems as well as for agricultural crops. These precious pollinators are known to be declining worldwide, and one major factor contributing to this decline are infections by parasites. Knowledge about parasites in wild bumblebee populations is thus of paramount importance for conservation purposes. We here report the geographical distribution of Crithidia and Nosema, two common parasites of bumblebees, in a yet poorly investigated country: Mexico. Based on sequence divergence of the Cytochrome b and Glycosomal glyceraldehyde phosphate deshydrogenase (gGPDAH) genes, we discovered the presence of a new Crithidia species, which is mainly distributed in the southern half of the country. It is placed by Bayesian inference as a sister species to C. bombi. We suggest the name Crithidia mexicana for this newly discovered organism. A population of C. expoeki was encountered concentrated on the flanks of the dormant volcanic mountain, Iztaccihuatl, and microsatellite data showed evidence of a bottleneck in this population. This study is the first to provide a large-scale insight into the health status of endemic bumblebees in Mexico, based on a large sample size (n=3,285 bees examined) over a variety of host species and habitats.

Trypanosomatids infecting honey bees have been poorly studied with molecular methods until recently. After the description of Crithidia mellificae (Langridge and McGhee, 1967) it took about forty years until molecular data for honey bee trypanosomatids became available and were used to identify and describe a new trypanosomatid species from honey bees, Lotmaria passim (Evans and Schwarz, 2014). However, an easy method to distinguish them without sequencing is not yet available. Research on the related bumble bee parasites Crithidia bombi and Crithidia expoeki revealed a fragment length polymorphism in the internal transcribed spacer 1 (ITS1), which enabled species discrimination. In search of fragment length polymorphisms for differential diagnostics in honey bee trypanosomatids, we studied honey bee trypanosomatid cell cultures of C. mellificae and L. passim. This research resulted in the identification of fragment length polymorphisms in ITS1 and ITS1-2 markers, which enabled us to develop a diagnostic method to differentiate both honey bee trypanosomatid species without the need for sequencing. However, the amplification success of the ITS1 marker depends probably on the trypanosomatid infection level. Further investigation confirmed that L. passim is the dominant species in Belgium, Japan and Switzerland. We found C. mellificae only rarely in Belgian honey bee samples, but not in honey bee samples from other countries. C. mellificae was also detected in mason bees (Osmia bicornis and Osmia cornuta) besides in honey bees. Further, the characterization and comparison of additional markers from L. passim strain SF (published as C. mellificae strain SF) and a Belgian honey bee sample revealed very low divergence in the 18S rRNA, ITS1-2, 28S rRNA and cytochrome b sequences. Nevertheless, a variable stretch was observed in the gp63 virulence factor.