Project description:This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/ Teladorsagia circumcincta, the brown stomach worm, is the most important gastro-intestinal parasite of sheep in temperate regions and is present on 100% of sheep farms in the UK. Infection with T. circumcincta causes a spectrum of disease in the host, ranging from weight loss to death, and results in significant production losses. Parasitic disease is currently controlled by treatment with anthelmintics but resistance to these drugs is widespread and threatens the viability of the livestock industry. There are now well-documented cases of UK farms harbouring triple-resistant T. circumcincta, which can no longer be controlled by any of the three major anthelmintic classes, forcing farmers to abandon sheep rearing. High-throughput sequencing of life stages of T. circumcincta will be used for gene finding and transcriptome analysis.
Project description:Gastrointestinal nematode (GIN) is a major economic and health concern is sheep farming. Sheep breeds such as Texel are relatively resistant to GIN than the Suffolk. With the objective to understand the underlying genetic mechanism of resistance and susceptibility at the transcriptomic level, two groups of animal from both the breed were artificially (orally) infected with 30,000 L3 larvae of prominent GIN Teladorsagia circumcincta. Subgroups of animals from each breed were slaughtered on day 0, 3, 7, 14 and 21 of post infection (p.i.). Transcriptomic profiling of abomasal lymph node was performed using RNA-seq. The perturbations in gene expression profiles in both the breeds were evident and Texel showed a more tightly regulated immune response than the Suffolk. The number of differentially expressed (DE) genes between the breeds was highest (437) on un-infected control (day 0) and lowest (173) on day 7 p.i.. Pathway analysis of DE genes identified 3 significant pathways, which involved only more highly expressed genes of Suffolk breed on day 0 and only more highly expressed genes of Texel (with one exception) on day 7 p.i.. The Th1, Th2 and Treg response was evident in response to GIN in Texel and was synchronized, while in Suffolk Th1 response was reduced after infection and pronounced Th2 and Treg was not evident. The study suggests maximum level of transcriptional activity in both breeds on day 7 p.i. and there was a shift of transcriptional activity from Suffolk on day 0 to Texel on day 7 p.i.. Suffolk had a reduced Th1 response with less pronounced Th2 and Treg immune response, while Texel had an active and synchronized Th1/Th2/Treg immune response in response to GIN infection. Abomasal lymph node tissue was taken from control (n=10) and experimentally infected (with T. circumcincta) lambs (n=36) from Texel and Suffolk breed on day 0, 3, 7, 14 and 21 post infection.
Project description:Gastrointestinal nematode (GIN) is a major economic and health concern is sheep farming. Sheep breeds such as Texel are relatively resistant to GIN than the Suffolk. With the objective to understand the underlying genetic mechanism of resistance and susceptibility at the transcriptomic level, two groups of animal from both the breed were artificially (orally) infected with 30,000 L3 larvae of prominent GIN Teladorsagia circumcincta. Subgroups of animals from each breed were slaughtered on day 0, 3, 7, 14 and 21 of post infection (p.i.). Transcriptomic profiling of abomasal lymph node was performed using RNA-seq. The perturbations in gene expression profiles in both the breeds were evident and Texel showed a more tightly regulated immune response than the Suffolk. The number of differentially expressed (DE) genes between the breeds was highest (437) on un-infected control (day 0) and lowest (173) on day 7 p.i.. Pathway analysis of DE genes identified 3 significant pathways, which involved only more highly expressed genes of Suffolk breed on day 0 and only more highly expressed genes of Texel (with one exception) on day 7 p.i.. The Th1, Th2 and Treg response was evident in response to GIN in Texel and was synchronized, while in Suffolk Th1 response was reduced after infection and pronounced Th2 and Treg was not evident. The study suggests maximum level of transcriptional activity in both breeds on day 7 p.i. and there was a shift of transcriptional activity from Suffolk on day 0 to Texel on day 7 p.i.. Suffolk had a reduced Th1 response with less pronounced Th2 and Treg immune response, while Texel had an active and synchronized Th1/Th2/Treg immune response in response to GIN infection.
Project description:Afferent Lymph was collected from up to 5 pen raised, nematode naïve, outbreed Romney sheep (Tag No’s 1001, 1002, 1003, 1004, 1006) over a 12 week period. During this time a Pre-Infection sample was collected, the sheep were immunised by 3 truncated (2 week) T. colubiformis infections (40,000 Tc) followed by a Challenge Infection with 40,000 T. colubiformis L3 (Challenge Infection weeks 1-3). The Challenge Infection will enable us to determine changes in gene expression when the sheep gut rejects the parasitic nematodes. The overall design is summarised in the below table.
Project description:Afferent Lymph was collected from up to 5 pen raised, nematode naïve, outbreed Romney sheep (Tag No’s 1001, 1002, 1003, 1004, 1006) over a 12 week period. During this time a Pre-Infection sample was collected, the sheep were immunised by 3 truncated (2 week) T. colubiformis infections (40,000 Tc) followed by a Challenge Infection with 40,000 T. colubiformis L3 (Challenge Infection weeks 1-3). The Challenge Infection will enable us to determine changes in gene expression when the sheep gut rejects the parasitic nematodes.
Project description:Tuft cells are required for an effective immune response to gastrointestinal parasitic nematodes in mice, but little is known about tuft cells in other animals. Using scRNA-seq, we profiled the transcriptome of tuft cells and other mucosal cell types from the sheep abomasal epithelium following nematode infection. Cells were isolated from abomasum mucosal tissue of two sheep and processed using 10x Genomics Chromium Single Cell 3’ Reagent kit (v.3) using a total of 20,000 input cells. cDNA libraries were sequenced on an Illumina NextSeq 500 system to a depth of 50,000 read pairs per cell. Sequences were mapped to the genomes for ovine (Ovis aries Oar_v3.1; https://www.ensembl.org/Ovis_aries/Info/Index) and bovine (Bos taurus ARS-UCD1.2; https://www.ensembl.org/Bos_taurus/Info/Index) and gene count matrices generated using the Cell Ranger software (v.3.1.0) and analysed using Seurat package (v.3.1) with R version 3.6.0. The “Find Markers” function identified a tuft cell cluster and other distinct epithelial (Epcam+) and immune (Ptprc+) cell clusters.
Project description:Anaplasma phagocytophilum is the causative agent of tick-borne fever (TBF) in ruminants and human, equine and canine granulocytic anaplasmosis. A. phagocytophilum modifies host gene expression and immune response. The objective of this work was to analyze differential gene expression in A. phagocytophilum-infected sheep using microarray hybridization and real-time RT-PCR in experimentally and naturally infected animals. Keywords: disease state analysis