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:Our objective was to investigate differences in gene expression between 24 parasite-resistant hair and 24 susceptible wool lambs to determine genetic mechanisms involved in resistance to H. contortus. Half of the animals of each breed were infected and sacrificed at 3 or 27 days post-infection; the remaining animals were uninfected controls. Breed differences in abomasum and abomasal lymph node tissue gene expression were assessed using bovine cDNA microarrays. Over 60 transcripts differed between breeds for each tissue and infection status. Genes differentially expressed between hair and wool sheep 3 days PI were assessed for gene function and mechanisms for greater immune cell infiltration, abomasal tissue repair, Th17 response, and anticoagulation were present in parasite-resistant hair sheep. By 27 days PI, hair sheep had greater expression of genes involved in gut motility, inflammatory cytokines, and cell proliferation and differentiation compared to wool sheep. Changes in these processes indicate Caribbean hair sheep have a stronger inflammatory response when infected with H. contortus which may facilitate the increased parasite resistance observed in these sheep.
Project description:Background: Gastrointestinal nematodes are a serious cause of morbidity and mortality in grazing ruminants. The major ovine defence mechanism is acquired immunity, which develops over time in response to infection. Nematode resistance varies both within and between breeds and is moderately heritable (h ~ 0.3). A detailed understanding of the genes and mechanisms involved in protective immunity, and the factors that regulate this response, is required to aid future breeding strategies as well as the development of effective and sustainable nematode control methods. The aim of this study was to compare the abomasal lymph node transcriptome of resistant and susceptible lambs in order to determine biological processes differentially expressed between resistant and susceptible lambs. Results: Scottish Blackface lambs, with divergent phenotypes for resistance, were challenged with 30,000 Teladorsagia circumcincta larvae (L3), and abomasal lymph node recovered at 7 and 14 days post-infection (dpi). High-throughput sequencing of abomasal lymph node cDNA was used to quantitatively sample the transcriptome with an average of 32 million reads per sample. A total of 194 and 144 genes were differentially expressed between resistant and susceptible lambs at 7 and 14 dpi respectively. Differentially expressed networks and biological processes were identified using Ingenuity Pathway Analysis. Resistant animals appear to generate a more rapid immune response as at 7 dpi processes relating to homing of lymphocytes, leukocyte migration and migration of antigen presenting cells were up-regulated. In susceptible animals this response appears to be delayed until approximately 14 dpi. Twenty-four Single Nucleotide Polymorphims (SNP), within 11 differentially expressed genes were tested for association with gastrointestinal nematode resistance in the Scottish Blackface lambs. Four SNPs in two genes (SLC30A2, and ALB) were suggestively associated with faecal egg count. Conclusions: A large number of genes were differentially expressed in the abomasal lymph node of resistant and susceptible lambs responding to gastrointestinal nematode challenge. Resistant Scottish Blackface lambs appear to generate a more rapid immune response to T. circumcincta. In susceptible lambs this response appears to be delayed until approximately 14 days post infection. SNP in two differentially expressed genes were suggestively associated with faecal egg count indicating that differentially expressed genes can be considered as candidate loci for mediating nematode resistance.
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:Haemonchus contortus is the gastrointestinal nematode (GIN) species that most parasites sheep and goats in tropical and subtropical regions. Due to the high prevalence and pathogenicity by hematophagy, it causes severe anemia, submandibular edema and deaths. Post-genomics tools, such as proteomics, allow the identification of differentially expressed genes and differentially abundant proteins and metabolites between two conditions of a given factor. Thus, the detection of protein profiles that occur in more parasitized animals may be of great interest for the identification of sheep that effectively need anthelmintic treatment. As a result, the parasites are preserved in refugia through Targeted Selective Treatment (TST) strategies for the animals, extension of the period of efficacy of the anthelmintics and, when its use is necessary, parasite control will be more efficient. This project aims to characterize the plasma proteomic profile of sheep breeds susceptible and resistant to GIN infection, aiming at the future development of a diagnostic tool and the understanding of the resistance mechanisms involved in the Santa Inês, White Dorper and Texel sheep breeds. In this way, the development of methodologies, processes and products for the identification of sheep with higher rates of infection by GIN, in addition to the detection of breeds/individuals more resistant to these parasites and the understanding of the resistance mechanisms involved in the different hosts, is presented as a topic of great relevance for sheep farming, which can be extended to goat farming. It also addresses public health issues and the consumer population, which is increasingly demanding in terms of quality, certification and safety.
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:The bryophyte was collected and dried. Aqueous extract was prepared and different concentration of the extracts were treat against Ascardia galli, chicken nematode and Trichostrongylus, sheep nematode. Control was maintained for both. The control and treated samples were further processed to study the gene expression by isolating and sequencing mRNA using Illumina next-gen sequencing.
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: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:This experiment analyzes the changes in expression of twelve days old Arabidopsis roots at ten hours post inoculation upon cyst nematode H. schachtii infection.