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.
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. Comparisons of control vs Nosema ceranae bees
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: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: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: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: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:Here, we examined the transcriptional and epigenetic (DNA methylation) responses to viral infection in honey bee workers. One-day old worker honey bees were fed solutions containing Israeli Acute Paralysis Virus (IAPV), a virus which causes muscle paralysis and death and has previously been associated with colony loss. Uninfected control and infected, symptomatic bees were collected within 20-24 hours after infection. Worker fat bodies, the primary tissue involved in metabolism, detoxification and immune responses, were collected for analysis. We performed transcriptome- and bisulfite-sequencing of the worker fat bodies to identify genome-wide gene expression and DNA methylation patterns associated with viral infection. There were 753 differentially expressed genes (FDR<0.05) in infected versus control bees, including several genes involved in epigenetic and antiviral pathways. DNA methylation status of 156 genes (FDR<0.1) changed significantly as a result of the infection, including those involved in antiviral responses in humans. There was no significant overlap between the significantly differentially expressed and significantly differentially methylated genes, and indeed, the genomic characteristics of these sets of genes were quite distinct. Our results indicate that honey bees have two distinct molecular pathways, mediated by transcription and methylation, that modulate protein levels and/or function in response to viral infections.
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.