Project description:Parasitic nematodes cause diseases that adversely impact on animal health and production. Although advances in genomics and transcriptomics are revolutionising the way we explore these parasites, there is a dearth of proteomic data to underpin or support fundamental discoveries. Here, using a high throughput LC-MS/MS-based approach, we undertook the first large-scale (global) proteomic investigation of Haemonchus contortus (the barber's pole worm), one of the most important parasitic nematodes of livestock animals worldwide. In total, 2487 unique H. contortus proteins, representing five key developmental stages [i. e. eggs, third-stage (L3) and fourth-stage (L4) larvae; female (Af) and male adults (Am)] were identified and quantified with high confidence. Bioinformatic analyses of the somatic proteome of H. contortus discovered substantial alterations in protein profiles during the life cycle, particularly in the transition from the free-living to the parasitic phase, and identified groups of key proteins involved specifically in feeding, digestion, metabolism, development, parasite-host interactions (including immunomodulation), structural remodelling of the body wall and other adaptive processes during the parasitic phase. This global proteomic data set will likely facilitate future molecular, biochemical and physiological investigations of H. contortus and related nematodes, and should underpin the discovery of novel intervention targets against haemonchosis.
Project description:We have investigated the transcriptomic response of the model nematode Caenorhabditis elegans to ivermectin (IVM); an important anthelmintic for human and animal parasite control. The transcriptomic response of the mutant strain DA1316 avr-14(ad1302); avr-15(ad1250); glc-1(pk54), which is highly resistant to ivermectin due to null mutations in three glutamate-gated chloride channel subunits, was examined. Despite the resistant nature of this strain, pharyngeal pumping rate was decreased following 4 hrs exposure to 100ng/ml and 1?g/ml ivermectin resulting in significant change in the expression level of genes associated with a fasting response. Matched cultures of synchronised C. elegans were grown to L4 stage on standard NGM plates with an OP50 bacterial lawn. The nematodes were then transferred to NGM plates containing 100ng/ml ivermectin, 1?g/ml ivermectin, or DMSO excipient only (control) for 4 hours. RNA was extracted from five biological replicates including controls for both the 100ng/ml ivermectin and 1?g/ml ivermectin experiments and hybridised to Affymetrix arrays.
Project description:Anthelmintic resistance is a major problem in the global fight against parasitic nematodes., Most previous studies have focused on the analysis of potential candidate genes that may have a role in resistance. Here we take a novel approach in the important parasite, Haemonchus contortus, by investigating changes in microRNA expression between resistant and susceptible parasites. The resistant worms included two geographically distinct strains and two lines generated by multiple rounds of backcrossing between susceptible and resistant parents, with ivermectin selection. All four resistant strains showed increased abundance of a single miRNA, hco-miR-9551, compared to the susceptible strain. hco-miR-9551 is enriched in female worms, is likely to be located on the X chromosome and is found only in clade V parasitic nematodes. Approximately 5-10% of the genomes of the resistant parental strains are introgressed into the respective backcrosses, suggesting that hco-miR-9551, or genes regulated by the miRNA, may be genetically linked to a locus that determines resistance. Genes containing predicted binding sites for hco-miR-9551 were identified computationally and refined based on differential expression in a transcriptomic dataset. These findings advance our conceptual understanding of the molecular mechanisms of anthelmintic resistance in H. contortus and indicate that altered miRNA expression may be linked with drug resistance. All samples were carried out using three biological replicates
Project description:We report analyses of the types, amounts and stage-dependence of microRNAsfound in D. immitis culture media recovered after incubating 800,000 microfilariae for 6 days, 500 third-stage larvae (L3) and 500 fourth-stage larvae (L4) for 7 days, as well as 40 adult females and 40 adult males for 48 hours, all separately.
Project description:To investigate the molecular changes (protein level) associated with dafachronic acid and the nuclear hormone receptor DAF-12 in Haemonchus contortus following exsheathment of the thrid-stage larvae, protein was extracted from replicates (n = 4), processed and analysed using LC-MS/MS.
Project description:Anthelmintic resistance is a major problem in the global fight against parasitic nematodes., Most previous studies have focused on the analysis of potential candidate genes that may have a role in resistance. Here we take a novel approach in the important parasite, Haemonchus contortus, by investigating changes in microRNA expression between resistant and susceptible parasites. The resistant worms included two geographically distinct strains and two lines generated by multiple rounds of backcrossing between susceptible and resistant parents, with ivermectin selection. All four resistant strains showed increased abundance of a single miRNA, hco-miR-9551, compared to the susceptible strain. hco-miR-9551 is enriched in female worms, is likely to be located on the X chromosome and is found only in clade V parasitic nematodes. Approximately 5-10% of the genomes of the resistant parental strains are introgressed into the respective backcrosses, suggesting that hco-miR-9551, or genes regulated by the miRNA, may be genetically linked to a locus that determines resistance. Genes containing predicted binding sites for hco-miR-9551 were identified computationally and refined based on differential expression in a transcriptomic dataset. These findings advance our conceptual understanding of the molecular mechanisms of anthelmintic resistance in H. contortus and indicate that altered miRNA expression may be linked with drug resistance.
2016-03-25 | GSE79542 | GEO
Project description:EMS derived Ivermectin/Moxidectin resistance mutants in C. elegans
| PRJNA768320 | ENA
Project description:16srRNA sequences of ivermectin and moxidectin challenged bacterial isolates
Project description:We have investigated the transcriptomic response of the model nematode Caenorhabditis elegans to ivermectin (IVM); an important anthelmintic for human and animal parasite control. The transcriptomic response of the mutant strain DA1316 avr-14(ad1302); avr-15(ad1250); glc-1(pk54), which is highly resistant to ivermectin due to null mutations in three glutamate-gated chloride channel subunits, was examined. Despite the resistant nature of this strain, pharyngeal pumping rate was decreased following 4 hrs exposure to 100ng/ml and 1μg/ml ivermectin resulting in significant change in the expression level of genes associated with a fasting response.