Download Adam Blanchard - National Sheep Association

Understanding the inflammatory process in ovine
footrot to inform rational vaccine design.
Adam Blanchard1, Tracey Coffey1, Richard Emes1,2, Jasmeet Kaler1,
1
3
3
1
Peers Davies , Gary Entrican , Sean Wattegedera , Sabine Tötemeyer
1School
of Veterinary Medicine and Science, University of Nottingham; 2Advanced Data Analysis Centre, University of
Nottingham; 3Moredun Research Institute, Penicuik, Edinburgh
Footrot in sheep
Objective 1: Identify diversity of D. nodosus and of
the local host response in sheep with no disease and
the different disease states of footrot
 Major impact on animal welfare & economy
 Large economic impact on farmers
 Dichelobacter nodosus is the bacterial
cause of underrunning footrot, other
bacteria, including Fusobacterium
necrophorum, may play a role in disease
severity
Host response
http://www.grrinninbear.com.au/vet/vsb_cartoons/0506_02Cartoons.html
Dichelobacter
nodosus
Skin damage
Fusobacterium necrophorum
Other bacteria?
D nodosus
Whole genome
sequencing
Microbiome of
interdigital space
Footrot
ID/Scald
Healthy
Host
transcriptomics
Bacterial community
Fusobacterium necrophorum?
Other bacteria?
qPCR of selected
host genes for
validation
Aims of the project
Diversity of
bacteria in
interdigital space
& tissues
Multi-Locus Sequence
Typing (MLST) scheme
Dissecting the mechanisms of
damage versus protection
(classical & expanded)
house
keeping genes
house keeping &
virulence genes
Diversity of
D. nodosus
Experimental design:
Increased understanding of
host/pathogen interactions
Objectives:
Underpin the design of a
safe,
efficacious
and
affordable vaccine that
meets the needs of the
international sheep industry
1.
Identify diversity of D. nodosus and of host response in sheep
2.
Modelling host pathogen interactions using complex organ culture
models
3.
Understanding host recognition and signalling in response to
footrot pathogens
Phase 1 July 2015- Feb 2016
 Development of MLST scheme based on >100 D. nodosus whole
genome sequences published recently
 Establishment of microbiome analysis pipeline based on preliminary
data
 Single flock sampling (swabs) to estimate return rate of D. nodosus
isolates from healthy, ID and footrot feet
Phase 2 March – May 2016
 Sampling UK flocks for isolation of D. nodosus and microbiome
analysis
 Collection of swabs & biopsies at abattoir to isolate D. nodosus,
determine microbiome and the local host response
Expected outcomes:
By studying the local immune defences in the sheep foot and how the
bacteria causing footrot are recognised by the immune system, in
addition to a complementary molecular analysis of the primary
pathogen associated with disease, researchers hope to explain
better how the clinical signs of footrot are caused and how the
immune system contributes to this.
The study will help to inform new approaches to disease
management and ultimately disease prevention.
Animal Health Research Club
In 2012 BBSRC established the Animal Health Research Club (ARC) to
bring together industry and the research community to support
research that improves our understanding of resistance to pests
and diseases in farmed animals.
13 company members who contribute to funding research and take
part in directing the Club's activities.
Objective 2: Modelling host pathogen interactions
using complex organ culture models - ‘sheep feet in
petri dish’
Freeze/
thawing
Epidermis Dermis
Ovine Epidermal
Keratinocytes
Skin equivalent
keratinocyte co-culture
Interdigital
Biopsy
Skin equivalent
Ovine Dermal
Fibroblasts
Skin equivalent
fibroblast co-culture
Ex vivo organ
culture model
Infection with D. nodosus strains
Histology
Gene expression of key
inflammatory markers
(Collaboration: C Mülling, University of Leipzig)
Acknowledgements:
Grant numbers BB/M012085/1 (University of Nottingham) and BB/M011941/1 (Moredun Research
Institute). GE and SW are also supported by the Scottish Government Rural and Environment Science
and Analytical Services (RESAS) Division.
www.nottingham.ac.uk/vet