Disease susceptibility and gut health in the wild: Determining interactions between diet, gut microbiome, and immunity
Lead Research Organisation:
University of Edinburgh
Department Name: Sch of Biological Sciences
Abstract
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Technical Summary
We will study wild house mice in a multivariate environment. We will trap mice for 2 months/year for 3 years over 2 96-trap grids. We will tag up to 600 mice allowing a cull of >240 mice. At first capture mice (day (d) -4 to -1) will be tagged, sexed and measured, blood and faecal samples collected for immunological, parasitological and microbiome analyses, and released at capture site. At next capture (d0-2) mice will be randomly allocated to 1 of 4 groups: diet supplementation (DS) plus control Ig (cIg), DS plus anti-IL4 treatment (11B11), no diet supplementation (NDS) plus cIg, NDS plus 11B11, intraperitoneally injected with 1.5mg of 11B11 or cIg, and released at their trap site. Food supplementation will be provided continuously via feeding stations, equipped with RFID loggers. Animals recaptured on d5-7 and/or 10-12 will receive further doses of 11B11 or cIg. 2 weeks after treatment start, recaptured animals will be culled.
Tissues will be taken for: endoparasite burden, age, serum antibody/cytokine levels, faecal IgA and inflammatory markers, gut microbiome composition. Proximal colon samples will be stored in:
(a) RNA later for qPCR/RNA-Seq
(b) methacarn for histopathology
(c) snap-frozen in OCT for immunohistochemistry
Each year, caecal contents from the DS and NDS cIg groups will be collected and frozen. Prior to faecal transplant, samples from mice which have entered feeders >8 times will be assessed by 16S rRNA sequencing for exclusion of outliers, and pooled within groups (>15 animals/group). GF mice will be gavaged with pooled faecal slurries selected from DS or NDS wild mice; one group of GF mice will receive a faecal slurry from SPF laboratory mice; faecal transplants will be allowed to stabilise over 3 weeks prior to low dose T.muris infection. Within each of the 3 faecal transplant groups, mice will receive 11B11 or cIg. Mice will be culled 21 days post-infection and analysed as above.
Tissues will be taken for: endoparasite burden, age, serum antibody/cytokine levels, faecal IgA and inflammatory markers, gut microbiome composition. Proximal colon samples will be stored in:
(a) RNA later for qPCR/RNA-Seq
(b) methacarn for histopathology
(c) snap-frozen in OCT for immunohistochemistry
Each year, caecal contents from the DS and NDS cIg groups will be collected and frozen. Prior to faecal transplant, samples from mice which have entered feeders >8 times will be assessed by 16S rRNA sequencing for exclusion of outliers, and pooled within groups (>15 animals/group). GF mice will be gavaged with pooled faecal slurries selected from DS or NDS wild mice; one group of GF mice will receive a faecal slurry from SPF laboratory mice; faecal transplants will be allowed to stabilise over 3 weeks prior to low dose T.muris infection. Within each of the 3 faecal transplant groups, mice will receive 11B11 or cIg. Mice will be culled 21 days post-infection and analysed as above.
