Project description:The zoonotic mosquito-borne filarial nematode Dirofilaria repens causes subcutaneous and ocular infections in dogs, cats and humans. Microfilariae (mf) are taken up by mosquitoes from infected vertebrate hosts which develop in the mosquito to the infectious third stage larvae (L3). These are transmitted to new vertebrate hosts and develop over two further moults to adult worms. The aims of the project were 1) the de novo sequencing and annotation of the D. repens genome and 2) comparative transcriptome analyses of the developmental stages mf and L3 as we hypothesized remarkable differences in gene expression between the different developmental stages. Genomic DNA was obtained from adult male D. repens. RNA was extracted from microfilariae from naturally infected dogs and from L3 produced in Aedes aegypti mosquitoes fed on blood spiked with microfi-lariae. The 99.59 MB genome was around 17% larger than that of the related species D. immitis (dog heartworm) and contained 8.9% fewer predicted genes (10,357). Around 1.8% of identified proteins (206/11,262) could not be mapped to D. immitis. Out of these, 6 (2.9 %) presented an ortholog in all other considered filarial nematodes (e.g. Loa loa) and Caenorhabditis elegans. A sig-nificantly higher number of D. repens proteins as compared to D. immitis mapped to the filarial nem-atode L. loa, reflecting the similarity in biology of D. repens and L. loa. A total of 876 genes were differentially expressed, of which 591 could be annotated in UniProtKB/Swiss-Prot. In particular, 155 genes with a UniProtKB/Swiss-Prot annotation to C. elegans and filarial nematodes were upregulat-ed in the L3 and 57 in the mf stage, respectively. Fifteen GO BP (Gene Ontology Biological Pro-cesses) were significantly enriched for the L3 group and 12 for the mf. These data provide first in-sight into the differential gene expression profiles of this filarial nematode and can serve for future investigations of metabolic processes and stage-specific diagnostics.
Project description:Wolbachia pipientis is an intracellular symbiotic bacterium found in insects and arthropods. Wolbachia can decrease the vectorial capacity for various pathogens, such as the dengue virus, in Aedes aegypti. The purpose of this study was to determine the effect of Wolbachia (wMel strain) on the vectorial capacity of Ae. aegypti for Dirofilaria immitis. We analyzed gene expression patterns by RNA-seq in addition to the D. immitis infection phenotype in Ae. aegypti infected with and without wMel. Four Ae. aegypti strains, MGYP2.tet, MGYP2, Liverpol (LVP)-Obihiro (OB), and LVP-OB-wMel (OB-wMel) were analyzed for transcriptome comparison in Malpighian tubule at 2 days post infection. The correlation between Wolbachia infection, D. immitis infection phenotype and immune-related genes expression in Ae. aegypti was investigated.
Project description:We sequenced total RNA from Dirofilaria immitis in order to generate the first tissue-specific gene expression profile of a filarial nematode and its Wolbachia endosymbiont.
Project description:We sequenced total RNA from Dirofilaria immitis in order to generate the first tissue-specific gene expression profile of a filarial nematode and its Wolbachia endosymbiont. Examination of transcript levels in 7 different Dirofilaria immitis tissues, in duplicate, using Illumina GAIIx.
Project description:Laodelphax striatellus is naturally infected with the Wolbachia strain wStri, which significantly increase the fecundity of its host. Wolbachia-infected females produce 30%–40% more eggs than Wolbachia-uninfected females. MicroRNAs (miRNAs) are a class of endogenous non-coding small RNAs that play critical roles in the regulation of gene expression at post-transcriptional level. Here we report the differentially expressed miRNAs between Wolbachia-infected and Wolbachia-uninfected strains of L. striatellus ovaries. Our data may be helpful to explore the molecular mechanisms by which Wolbachia increase the fecundity of Laodelphax striatellus.