TL;DR: A prospective study of a large scale migratory bee keeping operation using high-frequency sampling paired with comprehensive molecular detection methods, including a custom microarray, qPCR, and ultra deep sequencing established seasonal incidence and abundance of known viruses, Nosema sp.
Abstract: Honey bees (Apis mellifera) play a critical role in global food production as pollinators of numerous crops. Recently, honey bee populations in the United States, Canada, and Europe have suffered an unexplained increase in annual losses due to a phenomenon known as Colony Collapse Disorder (CCD). Epidemiological analysis of CCD is confounded by a relative dearth of bee pathogen field studies. To identify what constitutes an abnormal pathophysiological condition in a honey bee colony, it is critical to have characterized the spectrum of exogenous infectious agents in healthy hives over time. We conducted a prospective study of a large scale migratory bee keeping operation using high-frequency sampling paired with comprehensive molecular detection methods, including a custom microarray, qPCR, and ultra deep sequencing. We established seasonal incidence and abundance of known viruses, Nosema sp., Crithidia mellificae, and bacteria. Ultra deep sequence analysis further identified four novel RNA viruses, two of which were the most abundant observed components of the honey bee microbiome (∼1011 viruses per honey bee). Our results demonstrate episodic viral incidence and distinct pathogen patterns between summer and winter time-points. Peak infection of common honey bee viruses and Nosema occurred in the summer, whereas levels of the trypanosomatid Crithidia mellificae and Lake Sinai virus 2, a novel virus, peaked in January.
TL;DR: The novel results highlight that although secondary metabolites may not rescue survival in infected bees, they may play a vital role in mediating Crithidia transmission within and between colonies by reducingCrithidia infection intensities.
Abstract: The synthesis of secondary metabolites is a hallmark of plant defence against herbivores. These compounds may be detrimental to consumers, but can also protect herbivores against parasites. Floral nectar commonly contains secondary metabolites, but little is known about the impacts of nectar chemistry on pollinators, including bees. We hypothesized that nectar secondary metabolites could reduce bee parasite infection. We inoculated individual bumblebees with Crithidia bombi, an intestinal parasite, and tested effects of eight naturally occurring nectar chemicals on parasite population growth. Secondary metabolites strongly reduced parasite load, with significant effects of alkaloids, terpenoids and iridoid glycosides ranging from 61 to 81%. Using microcolonies, we also investigated costs and benefits of consuming anabasine, the compound with the strongest effect on parasites, in infected and uninfected bees. Anabasine increased time to egg laying, and Crithidia reduced bee survival. However, anabasine consumption did not mitigate the negative effects of Crithidia, and Crithidia infection did not alter anabasine consumption. Our novel results highlight that although secondary metabolites may not rescue survival in infected bees, they may play a vital role in mediating Crithidia transmission within and between colonies by reducing Crithidia infection intensities.
TL;DR: Thymidylate synthetase and dihydrofolate reductase exist as a bifunctional protein in a number of species of protozoa which span diverse groups of the subkingdom and it is likely that the bifunctionsal protein is widespread among these primitive eukaryotes.
TL;DR: Analysis of the characteristics of protein import into mitochondria of Trypanosoma brucei suggests a resemblance between the import machineries of mitochondria and hydrogenosomes.
TL;DR: Dihydrofolate reductase and thymidylate synthase activities of the rodent malaria organism Plasmodium berghei also copurified on Sephadex G-200 and methotexate-Sepharose columns, suggesting that this unique bifunctional protein might occur throughout the Protozoa.
Abstract: The molecular weight of dihydrofolate reductase (5,6,7,8-tetrahydrofolate:NADP+ oxidoreductase, EC 1.5.1.3) from protozoa has been reported to be 5- to 10-fold larger than the isofunctional enzyme of most other organisms studied, based on gel filtration. This enzyme from the protozoal flagellate Crithidia fasciculata has been purified to homogeneity and found to be a bifunctional protein with thymidylate synthase (5,10-methylene tetrahydrofolate:dUMP C-methyltransferase, EC 2.1.1.45) activity. The purified protein, eluted from methotrexate-Sepharose columns by dihydrofolate, migrated as a single band on both nondenaturing and denaturing polyacrylamide gel electrophoresis. The monomer Mr is 56,700 +/- 200. The native Mr was calculated to be 107,000 from a sedimentation coefficient of 5.9 and Stokes radius of 4.4 nm. Dihydrofolate reductase and thymidylate synthase activities of the rodent malaria organism Plasmodium berghei also copurified on Sephadex G-200 and methotexate-Sepharose columns, suggesting that this unique bifunctional protein might occur throughout the Protozoa.