The phylogenetic relationship between populations of marginally and sympatrically located Pinus halepensis Mill. and Pinus brutia Ten. in Turkey, based on the ITS-2 region

Turkish red pine (Pinus brutia) is a widespread and important forest tree species in Turkey, occurring mainly in southern, western, and northwestern Turkey, while the natural occurrence of Aleppo pine (Pinus halepensis) is restricted to 2 locations and is found sympatrically with Turkish red pine. In the present study sympatric populations of both species from Mugla and Adana provinces in Turkey were sampled, and the internal transcribed spacer 2 (ITS-2) region of ribosomal DNA was comparatively studied with sequence analysis. Analysis of molecular variance (AMOVA) demonstrated 100% of total molecular variation between the species in Mugla province, versus only 50.65% in Adana province. Construction of a phylogenetic tree with a bootstrap value of 92% revealed that Aleppo pine and Turkish red pine samples at the species level were well separated. Estimated F, values indicated that Turkish red pine and Aleppo pine were highly differentiated in Mugla province due to possible reproductive isolation, while the 2 species shared a more common genetic background due to possible natural hybridization in Adana province

Differential diagnosis of the honey bee trypanosomatids Crithidia mellificae and Lotmaria passim

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

Differential diagnosis of the honey bee trypanosomatids Crithidia mellificae and Lotmaria passim

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.publisher: Elsevier articletitle: Differential diagnosis of the honey bee trypanosomatids Crithidia mellificae and Lotmaria passim journaltitle: Journal of Invertebrate Pathology articlelink: http://dx.doi.org/10.1016/j.jip.2015.06.007 content_type: article copyright: Copyright © 2015 Elsevier Inc. Published by Elsevier Inc. All rights reserved.status: publishe

Evaluation Of Cage Designs And Feeding Regimes For Honey Bee (Hymenoptera: Apidae) Laboratory Experiments

The aim of this study was to improve cage systems for maintaining adult honey bee (Apis mellifera L.) workers under in vitro laboratory conditions. To achieve this goal, we experimentally evaluated the impact of different cages, developed by scientists of the international research network COLOSS (Prevention of honey bee COlony LOSSes), on the physiology and survival of honey bees. We identified three cages that promoted good survival of honey bees. The bees from cages that exhibited greater survival had relatively lower titers of deformed wing virus, suggesting that deformed wing virus is a significant marker reflecting stress level and health status of the host. We also determined that a leak- and drip-proof feeder was an integral part of a cage system and a feeder modified from a 20-ml plastic syringe displayed the best result in providing steady food supply to bees. Finally, we also demonstrated that the addition of protein to the bees' diet could significantly increase the level of vitellogenin gene expression and improve bees' survival. This international collaborative study represents a critical step toward improvement of cage designs and feeding regimes for honey bee laboratory experiments.WoSScopu