Trancriptome analysis of honey bees (Apis mellifera) infected with Nosema ceranae
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ABSTRACT: The microsporidia Nosema ceranae are intracellular parasites that proliferate in the midgut epithelial cells of honey bees (Apis mellifera). To analyze the pathological effects of those microsporidia, we orally infected honey bee workers 7 days after their emergence. Bees were flash frozen 15 days after the infection. Then, the effects on the gut ventriculi were analyzed and compared to non-infected (control) bees. Comparisons of control vs Nosema ceranae bees
Project description:In this study we addressed whether the transcriptome profile in the honey bee brain is similar for two major parasites of honey bee, Varroa destructor and Nosema ceranae. Honey bees parasitized by these two parasites show accelerated behavioral maturation and deficiences in orientation and learning/memory that we hoped to characterized at the transcriptomic level. honey bee adults infested by Varroa destructor or Nosema ceranae compared to control bees, in duplicate
Project description:Experimental infection of (2 days old) adult honey bee workers (30 bees per replicates, 3 replicates per treatments, from 3 different colonies (one colony per cage for each treatment)) with 10^9 genome equivalent of Black Queen Cell Virus (BQCV) in 10µl of sugar solution and/or 10^5 fresh Nosema ceranae spores (control bees were given a similar bee extract in PBS, without pathogen). Bees were kept in cages of 30 bees in incubator (30°C/50%RH). At day 13 p.i., bees were flash frozen, and stored at -80°C. Brain mRNA profiles of 15 old bees were generated by deep sequencing, in triplicates except for bees infected by both Nosema ceranae and Black Queen Cell Virus (duplicates)
Project description:The microsporidia Nosema ceranae are intracellular parasites that proliferate in the midgut epithelial cells of honey bees (Apis mellifera). To analyze the pathological effects of those microsporidia, we orally infected honey bee workers 7 days after their emergence. Bees were flash frozen 15 days after the infection. Then, the effects on the gut ventriculi were analyzed and compared to non-infected (control) bees.
Project description:Bees from 3 unrelated colonies were injected with 1ul PBS extract containing 10^9 genome equivalent of Deformed wIng virus (DWV) and/or fed 10µl sucrose solution containing 10^5 fresh Nosema ceranae spores. Control bees were injected and fed with an equivalent DWV- and Nosema-free extract respectively). Bees were kept in cages of 21 bees (7 from each colony), and each cage was replicated 5 times per each of the 4 treatments). Bess were kept in an incubator at 30°C/50%RH. At day 12 p.i., bees were flash frozen in liquid nitrogen, and stored at -80°C. Bee abdomen RNA was sent to Christina Grozinger lab (Penn State, USA). RNA was pooled for 3 abdomens per replicate for 5 replicates per treatment. Arrays were hybridized in a dye-swap loop design.
Project description:In this study we addressed whether the transcriptome profile in the honey bee brain is similar for two major parasites of honey bee, Varroa destructor and Nosema ceranae. Honey bees parasitized by these two parasites show accelerated behavioral maturation and deficiences in orientation and learning/memory that we hoped to characterized at the transcriptomic level.
Project description:Experimental infection of (2 days old) adult honey bee workers (30 bees per replicates, 3 replicates per treatments, from 3 different colonies (one colony per cage for each treatment)) with 10^9 genome equivalent of Black Queen Cell Virus (BQCV) in 10µl of sugar solution and/or 10^5 fresh Nosema ceranae spores (control bees were given a similar bee extract in PBS, without pathogen). Bees were kept in cages of 30 bees in incubator (30°C/50%RH). At day 13 p.i., bees were flash frozen, and stored at -80°C.
Project description:We aim to evaluate the effects of four Nosema spores’ isolates, (i) and (ii) N. ceranae isolated from A. mellifera hosts from two different geographical origins, (iii) N. ceranae from A. cerana host and (iv) N. apis from A. mellifera, on the A. mellifera on gut proteomics at the early stage of infection. To dissect the molecular mechanism responsible of the susceptibility of A. mellifera to Nosema, we investigated by high-resolution proteomics (LC-ESI-MS/MS) and differential label-free quantification of proteins (LFQ) the molecular cross-talk existing between different species and isolates of N. apis and N. ceranae, and the targetted gut tissue of A. mellifera. To reach the objectives of this study, we performed a bottom-up proteomic analysis on the different anatomical sections of the gut tissue (esophagus, crop, midgut, ileum and rectum) at an early stage of the exposition to Nosema spores (4 days). Then, we focused on the midgut, the region targeted by Nosema sposres for germination and, as we found out, the second region with the highest load of Nosema proteins, after the rectum, to perform differential quantitative proteomic analyses and acquire series of up- and down-regulated proteins. We discussed the different pathways observed to be impacted by different Nosema species and isolates with a main focus on the deregulated metabolic and response to stimuli processes.
Project description:Newly emerged adult workers (24 hours old) were infected with 25,000 Nosema apis spores and 25,000 Nosema ceranae spores in sucrose solution. Controls were fed sucrose. Workers were maintained in cages in an incubator and collected at 14 days post-infection. Fat bodies (eviscerated abdomens) were dissected and whole genome expression in this tissue was compared across treatments using microarrays.
Project description:The majority of fungal species prefer the 12°–30° C range and relatively few species tolerate temperatures higher than 35° C . Our understanding of the mechanisms underpinning the ability of some species to grow at higher temperatures is incomplete. Nosema ceranae is an obligate intracellular fungal parasite that infects honey bees and can cause individual mortality and contribute to colony collapse. Despite a reduced genome, this species is strikingly thermotolerant, growing optimally at the colony temperature of 35° C. In characterizing the HSR in N. ceranae, we found that this and other microsporidian species have lost the transcriptional regulator HSF and possess a reduced set of putative core HSF1-dependent HSR target genes. Despite these losses, N. ceranae demonstrates robust upregulation of the remaining HSR target genes after heat shock. In addition, thermal stress leads to alterations in genes involved in various metabolic pathways, ribosome biogenesis and translation, and DNA repair. These results provide important insight into the stress responses of microsporidia. Such new understanding will allow new comparisons with other pathogenic fungi and potentially enable discovery of novel treatment strategies for microsporidia infections affecting food production and human health.