Mapping triclabendazole resistance in Fasciola hepatica using next generation sequencing technologies

Lead Research Organisation: University of Liverpool
Department Name: Veterinary Pathology

Abstract

Fasciolosis is a common and important disease of livestock. In the UK it is the most commonly reported infection associated with the digestive tract of ruminants. It is caused by Fasciola hepatica, a flatworm found in the liver and transmitted by a mud snail. Fasciolosis is controlled predominantly using drug treatment and the most commonly used drug is triclabendazole (TCBZ). This is the only drug that is effective against the immature stages of the parasite that are responsible for acute, often fatal disease. It is also the drug of choice for human fasciolosis, caused by the same parasite. Resistance to TCBZ was first reported in Australia in 1995 and is now thought to be widespread worldwide. Very little is known about mechanisms of drug resistance in F. hepatica, which genes are involved and how resistance is evolving. This proposal aims to exploit our ability to create clones of resistant and susceptible F. hepatica; to derive genetically identical infections of TCBZ-susceptible and TCBZ-resistant isolates in sheep, from which we will generate second generation populations. These populations will be used to identify markers, in this case single mutations at the DNA level that associate with resistance genes during the crossing of resistant and susceptible clones. We will use rapid, high throughput, next generation sequencing technologies, which allow us to produce a preliminary F. hepatica genome map. The resistance markers will be mapped onto this preliminary genome and allow us to identify regions of the genome where the gene or genes responsible for encoding resistance are found. At the end of the project we will have genetic markers for resistance that could be used to detect and track the emergence of resistance in populations of F. hepatica in naturally infected sheep and cattle in the UK, but, importantly they will also be used in future projects to start to home in and identify genes encoding resistance. There will be other important outputs for the wider scientific community from the project, such as a bank of genome data for the parasite and clones of the parasite whose resistance status is well characterised. Genome data will be made available through a publicly accessible website and the parasite clones will be made available to other groups by lodging them with our industrial partner, Ridgeways Research Ltd.

Technical Summary

Fasciola hepatica causes serious disease in livestock worldwide. It is common in the UK, its prevalence is increasing and the costs of controlling infection have a significant impact on the UK agricultural sector. Resistance to triclabendazole (TCBZ), the most widely used drug for the control of fasciolosis, is reported in the UK, Europe and Australia. This project will take a new approach to identifying markers for resistance that will, in the long term, be useful in the identification of gene(s) encoding resistance and the development of assays to measure the prevalence and spread of resistance. Clonal lines will be generated from resistant and susceptible field isolates and F1 and F2 generations created based on crosses of these lines. A draft genome sequence for F. hepatica will be created using 454 fragment and mate pair libraries at 10x coverage producing a rough assembly (N50 of 30kb) complemented by Solexa reads and the MAKER pipeline used for basic annotation. Resistant and susceptible clonal lines will be resequenced at 20x coverage to identify single nucleotide polymorphisms (SNP) using SOLiD fragment libraries. Differences in SNP allele frequencies will be identified between parasites that have developed after exposure to TCBZ versus unexposed controls. Six pairs of treated vs control populations will be sequenced at 20x coverage using SOLiD fragment libraries. Allele frequencies will be analyzed using logistic regression plotted against the draft genome sequence to identify genomic regions associated with TCBZ resistance. Finally a sample of 150 SNPs linked to TCBZ resistance, plus 100 unlinked controls will be validated. Individual miracidia will be SNP genotyped to determine linkage between markers to indicate if a single locus is responsible for resistance and if TCBZ resistance is a dominant or recessive trait. Finally experimental infections using F2 individuals will be used to confirm linkage of SNPs to TCBZ resistance in vivo.

Planned Impact

The work we are proposing uses the latest genomic technologies combined with knowledge and exploitation of the life cycle of the parasite to elucidate the molecular mechanisms of triclabendazole resistance. The innovative nature of the work is only possible because of the recent advances in genome sequencing, but the work is necessarily technically complex and long term. In the short term the outputs of the project will be the identification of markers for resistance genes. This will lead to further research aimed at identifying specific genes of interest using the parasite material generated in this project, which, combined with the likely advances in sequencing and annotation technologies, will lead to identification of genes encoding resistance. From this we can start to understand the biochemical pathways associated with resistance, leading to ways to prevent resistance occurring - such as through modifications to the drug itself. Thus the tangible outputs and impact of the project are necessarily long term, but will ultimately lead to improved application of drug, markers for resistance and circumventing resistance mechanisms developed by the parasite. Markers for resistance will form the basis of sensitive and specific diagnostic tests for resistance in the field and ultimately provide evidence based recommendations for anthelmintic use. The principle, immediate beneficiaries from this work is the scientific community in general and specifically groups working on anthelmintic resistance. The new approach we are taking will provide information to groups working on related systems. The majority of molecular studies on anthelmintic resistance have focused on possible associations of putative candidate genes with a resistance phenotype. What we are proposing represents a timely and novel molecular approach using a combination of genetic and genomic resources to address drug resistance at a genome-wide level. As such, the work will have important outputs, useful to other academics, both those working directly in fluke research but also those working on other parasites and in the drug resistance field as a whole. Those groups working on the development of vaccines, the immunology and pathogenesis of fluke and improved diagnosis will benefit from free access to the partly annotated fluke genome. Fluke research will be greatly advance by having access to new, fully validated parasite clones. These will be made available through our commercial collaborator, Ridgeways Research Ltd. In the long term, veterinarians, farmers and food retailers will benefit from diagnostics for triclabendazole resistance, improved, strategic use of triclabendazole leading to reduced use of drug, which ultimately will lead to reduced drug residues in food and improve the safety of meat and milk products for the whole community.

Publications

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Beesley NJ (2018) Fasciola and fasciolosis in ruminants in Europe: Identifying research needs. in Transboundary and emerging diseases

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Cwiklinski K (2015) Characterisation of a novel panel of polymorphic microsatellite loci for the liver fluke, Fasciola hepatica, using a next generation sequencing approach. in Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases

 
Description Fasciolosis is a common and important disease of livestock in the UK. It has substantially increased in prevalence over the last decade and poses a great threat to livestock health and welfare. It is caused by the parasite Fasciola hepatica, a flatworm found in the liver and transmitted by a mud snail. Fasciolosis is controlled predominantly using drug treatment and the most commonly used drug is triclabendazole (TCBZ). This is the only drug that is effective against the immature stages of the parasite that are responsible for acute, often fatal disease. It is also the drug of choice for human fasciolosis, caused by the same parasite. Resistance to TCBZ was first reported in Australia in 1995 and is now thought to be widespread worldwide. Very little is known about the mechanisms of drug resistance in F. hepatica, which genes are involved and how resistance develops and spreads.
This proposal has exploited our ability to manipulate the parasite life cycle to create clones of resistant and susceptible F. hepatica; to derive infections of genetically identical TCBZ-susceptible and TCBZ-resistant isolates in sheep, from which we produced second generation populations. These populations were used to identify markers in the genome of F. hepatica that associate with resistance.

Our major findings are:

1) The first ever draft genome for Fasciola hepatica and production of a panel of markers throughout the genome, genome-wide single nucleotide polymorphisms or SNPs, that allow us to identify large sections of the genome.

2) Production and maintenance of six clonal isolates of triclabendazole-susceptible (TCBZ-S; n=3) and -resistant (TCBZ-R; n=3) isolates of F. heptica (each one derived from a from a single parasite).

3) By taking a clonal TCBZ-S parasite and allowing it to mate with a TCBZ-R F. hepatica parasite we have produced offspring, F2 recombinants, that have both TCBZ-resistance and -susceptibilty genes. These F2 offspring were treated with the drug TCBZ and only a proportion of parasites survived treatment. For the first time this shows that TCBZ resistance has a genetic basis and is heritable.
4. By comparing the frequency of genome-wide SNP markers in surviving parasites (TCBZ-resistant) to the same population in control (non-drug treated) samples (both TCBZ-resistant and susceptible) using a pooled genotyping approach we have localised regions of the genome associated with TCBZ resistance.

5. We have determined that Fasciola hepatica in the UK are diploid and exist as a panmictic (random mingling and mating when breeding) population, exhibiting high gene flow (transfer of genes from one generation to another). This suggests that once present resistance genes can move rapidly through liver fluke populations and even animals that are not frequently exposed to drug treatment can harbour resistant parasites.

6. We have shown that the snail intermediate host is capable of being infected with more than one genotype of F. hepatica and that may explain the high levels of diversity we see in the definitive host. This is preliminary work but in future it will help us understand the role of the snail in the transmission of drug resistant parasites.

7. Our recently published study has identified a number of sheep farms where triclabendazole resistance is present, assisting farmers in making treatment decisions.
Exploitation Route The genome dataset was in much demand from the international research field and was made available as an advanced draft in 20013 for six research laboratories worldwide and one industry partner.

The clonal isolates generated during this project have been shared with three other research groups and are publically available from our laboratories.

The candidate genes we are associating with TCBZ resistance will be of interest to a number of laboratories worldwide.

The panel of microsatellite and genome-wide SNPs we have generated are in use by our laboratory and will be useful to other researchers worldwide in order to better understand F. hepatica population structure, transmission dynamics and the influence of drug selection.

Metacercariae are vital to perform in vitro and in vivo studies of F. hepatica, to advance our knowledge of new drugs, vaccines and to mitigate against drug rapid resistance development. They are in short supply and there is a global demand for metacercariae that cannot be met currently. We supply metacercariae to groups and colleagues in collaboration.
Sectors Agriculture, Food and Drink,Education,Environment,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

URL http://www.liverpool.ac.uk/liver-fluke
 
Description Our Fasciola hepatica genome dataset: This has been used by a number of internationally-recognised research groups to identify genes that are candidate drug resistance genes, novel anthelmintic targets and vaccine candidates. We were approached by several research groups requesting access to our genome draft at an early point in the project. We provided access to an advanced draft to a number of groups and it substantially improved their ability to explore their gene families of interest. This led to collaborative projects and publications. Genetically defined clonal isolates of known triclabendazole sensitivity: These isolates have ensured that in vitro assays for drug sensitivity could be validated and used to phenotype populations of parasites ex vivo. They have also been used to explore the reproductive behaviour of F. hepatica in vivo. The work describing phenotypic and genotypic characterisation of these isolates has been published in 2018. Added value: Our panel of microsatellites generated have been applied to address the question of population structure in F. hepatica. This is the basis of an University of Liverpool/EU funded PhD student project. PhD submitted February 2016. A collaborative project with groups in Spain and Argentina, genotyping liver fluke isolates from livestock in Argentina has been completed and a manuscript is in preparation. Improving Diagnostics Complementary field studies identifying TCBZ-resistant F. hepatica populations have been carried out by a Malaysian Government-funded PhD student in our laboratory. She has validated diagnostic tools using archived faecal and serum samples from sheep experimentally infected with F. hepatica during the course of this project. PhD successfully awarded August 2015 and one publication has arisen from this work with a second in preparation. We have received funding from the Agriculture and Horticulture Development Board (AHDB) Beef and Lamb for a studentship. The project uses our genome dataset and archived sera from our experimentally infected animals to develop a diagnostic to detect acute infection with F. hepatica. We have a BBSRC DTP studentship that has been shortlisted and we have identified a high quality candidate for the project who is going to go forward to the final selection process. The project will explore the role of the snail intermediate host in the transmission of parasite genotypes and the potential for aggregation and long-term survival of metacercariae on pasture and in animal fodder. It is a CASE award with the AHDB Beef and Lamb industrial partner.
First Year Of Impact 2015
Sector Agriculture, Food and Drink,Education,Environment,Pharmaceuticals and Medical Biotechnology
Impact Types Societal,Economic

 
Description Provides an evidence base for advise for farmers via SCOPS and COWS
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
Impact Farmers, in particular sheep farmers lose livestock to fasciolosis every year. These data on triclabendazole resistance have fed into stakeholder and farmer lay texts, sustainable parasite control advice e.g SCOPS (https://www.scops.org.uk/) and raised awareness of the problem of drug resistance in liver fluke and has lead to changing behaviour. This is being developed as a REF 2020 impact case.
 
Description BBSRC DTP Studentship Scheme
Amount £74,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2016 
End 09/2020
 
Description BBSRC response Mode January 2016
Amount £812,000 (GBP)
Funding ID BB/P001912/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 03/2017 
End 06/2020
 
Description BBSRC:Research Grant United States-UK partnering award
Amount £47,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 05/2016 
End 04/2018
 
Description Institute of Infection and Global health Studentship scheme
Amount £60,000 (GBP)
Organisation University of Liverpool 
Sector Academic/University
Country United Kingdom
Start 10/2012 
End 09/2015
 
Title Clonal isolates of triclabendazole-susceptible and -resistant Fasciola hepatica 
Description Six genetically defined clonal isolates of Fasciola hepatica of known triclabendazole resistance status. These were produced from experimental infections of laboratory adapted snails using a single infective stage (miracidia) taken, primarily from infections representing the curent UK field situation in a number of geographical regions. Triclabendazole-resistant Fasciola hepatica a)FhepLivIsoR1 - NW England b)FhepLivIsoR2 - Wales c)FhepLivIsoR3 - NW England Triclabendazole-susceptible Fasciola hepatica d)FhepLivIsoS1 - Shrewsbury lab strain, commercially available susceptible isolate maintained by Ridgeway Research Ltd. e)FhepLivIsoS2 - Organic farm NW England f)FhepLivIsoS3 - Organic farm SW England 
Type Of Material Biological samples 
Year Produced 2013 
Provided To Others? Yes  
Impact International Anthelmintics Symposium. February 2014. McCammick E et al. , Queens University Belfast. The ABC's of Fasciola hepatica. 
 
Title Fasciola hepatica genome 
Description 1.25GB genome for Fasciola heptica - the first of its kind 
Type Of Material Technology assay or reagent 
Year Produced 2013 
Provided To Others? Yes  
Impact Enhanced the work of other research groups leading to multiple publications - greatly facilitated by ear;y release of genome prior to publication via the International consortium. Better understanding of Fasciola hepatica biology and facilitation of drug and vaccine design 
URL http://parasite.wormbase.org/Fasciola_hepatica_prjeb6687/Info/SpeciesLanding/
 
Title Panel of microsatellites for Fasciola hepatica 
Description A novel panel of microsatellite markers for genotyping F. hepatica populations/isolates. 
Type Of Material Technology assay or reagent 
Year Produced 2015 
Provided To Others? No  
Impact Characterisation of a novel panel of polymorphic microsatellite loci for the liver fluke, Fasciola hepatica, using a next generation sequencing approach.Krystyna Cwiklinski, Katherine Allen, James LaCourse, Diana J Williams, Steve Paterson and Jane E Hodgkinson. Under review. Infection, Genetics and Evolution. 
 
Title Production of metacercariae to meet researcher needs - development for commercial provision 
Description During the course of the project we maintained snail stocks and produced metacercariae (infective stage of Fasciola hepatica) for personal use. We also genotyped large numbers of liver fluke in the definitive host (sheep and cattle). There is a need to produce metacercariae that are representative of field isolates for both drug and vaccine trials but there is currently no such resource. We currently produce wild-type metas and we wish to start to produce them on a commercial scale. Unfortunately the BBSRC BBR proposal we wrote to underpin and develop this resource was not funded, despite 3 exceptional reviews. This resource is invaluable and in much demand so we will apply for a pathfinder grant to secure the resource in the longer term. 
Type Of Material Biological samples 
Year Produced 2015 
Provided To Others? Yes  
Impact Has allowed in vitro and in vivo work on developing new drugs and vaccines to go ahead as, since the major provider of metacercariae in the United States retired, they are in very short supply and threaten the future of liver fluke research meeting its maximum potential. Multiple publications have been facilitated by liver fluke research groups. 
 
Title Fasciola hepatica draft genome and a panel of genome-wide SNPs 
Description Advanced drafts of the F. hepatica genome were made available to a number of researchers throughout the lifetime of the project. These researchers include many of the most important researchers in the field of F. hepatica worldwide and constitute a consortium of fluke genomics. The dataset was made publically available in 2014 via the European Nucleotide Archive (ENA), European Molecular Biology Laboratory, European Bioinformatics Institute (EMBL-EBI), Wellcome Trust Genome Campus, Cambridge, UK. The Fasciola hepatica genome is available from the European Nucleotide Archive under accession numbers PRJEB6687 (read data) and ERZ021912 (assembly). The European Nucleotide Archive (ENA) provides a comprehensive record of the world's nucleotide sequencing information, covering raw sequencing data, sequence assembly information and functional annotation. Other associated files are available from the Centre for Genomic Research server (http://www.cgr.liv.ac.uk:8088/illum/Fasciolav2_db1af35899c4eb29/) and will soon be made available as part of the BBSRC-funded parasite-WormBase portal from EBI/Sanger Institute. 
Type Of Material Database/Collection of data 
Year Produced 2012 
Provided To Others? Yes  
Impact Cwiklinski, K., Dalton, J. P., Dufresne, P. J., La Course, J., Williams, D. J., Hodgkinson, J., & Paterson, S. (2015). The Fasciola hepatica genome: gene duplication and polymorphism reveals adaptation to the host environment and the capacity for rapid evolution. Genome Biology, 16: e71. Cwiklinski, K., Allen, K., LaCourse, J., Williams, D. J., Paterson, S., & Hodgkinson, J. E. (2015). Characterisation of a novel panel of polymorphic microsatellite loci for the liver fluke, Fasciola hepatica, using a next generation sequencing approach. Infection Genetics and Evolution, 32, 298-304. doi:10.1016/j.meegid.2015.03.014 Japa O, Hodgkinson JE, Emes RD, Flynn RJ. (2015). TGF-ß superfamily members from the helminth Fasciola hepatica show intrinsic effects on viability and development. Vet Res. 46:29. doi: 10.1186/s13567-015-0167-2 
URL http://www.cgr.liv.ac.uk:8088/illum/Fasciolav2_db1af35899c4eb29/
 
Description Collaboration to characterise Fasciola hepatica isolates from the Southern hemisphere (Australia) and compare with our UK characterisation 
Organisation La Trobe University
Country Australia 
Sector Academic/University 
PI Contribution My expertise in genotyping liver fluke and analysing data. Intellectual input. Training of staff. Access to equipment or facilities. Co-Investigator on a grant submission to Australian Research Council (ARC). Reviewed application and contributed to experimental design. Provided CV, publication and impact statement to ARC Commitment to support genotyping of isolates and detection of drug resistance alleles.
Collaborator Contribution Two principal investigators: Professor Terry Spithill, LaTrobe University and Dr Neil Young, University of Melbourne. Dr Young is the primary investigator (PI) and developed the ideas in collaboration with Prof Spithill and myself. Their expertise, intellectual input or the training of staff. Access to data, equipment or facilities. It is a 3 year project to support 3 PhD students and will involve work at both the University of Melbourne and University of Liverpool. The PI drafted the proposal outline and experimental objectives in consultation with Co-Is. If successful will involve transfer of parasite material for genetic analysis. Parasites will be collected by collaborators.
Impact Grant submission to ARC in February 2017. Currently under review.
Start Year 2016
 
Description Collaboration to characterise Fasciola hepatica isolates from the Southern hemisphere (Australia) and compare with our UK characterisation 
Organisation University of Melbourne
Country Australia 
Sector Academic/University 
PI Contribution My expertise in genotyping liver fluke and analysing data. Intellectual input. Training of staff. Access to equipment or facilities. Co-Investigator on a grant submission to Australian Research Council (ARC). Reviewed application and contributed to experimental design. Provided CV, publication and impact statement to ARC Commitment to support genotyping of isolates and detection of drug resistance alleles.
Collaborator Contribution Two principal investigators: Professor Terry Spithill, LaTrobe University and Dr Neil Young, University of Melbourne. Dr Young is the primary investigator (PI) and developed the ideas in collaboration with Prof Spithill and myself. Their expertise, intellectual input or the training of staff. Access to data, equipment or facilities. It is a 3 year project to support 3 PhD students and will involve work at both the University of Melbourne and University of Liverpool. The PI drafted the proposal outline and experimental objectives in consultation with Co-Is. If successful will involve transfer of parasite material for genetic analysis. Parasites will be collected by collaborators.
Impact Grant submission to ARC in February 2017. Currently under review.
Start Year 2016
 
Description Genotyping liver fluke from Argentina in collaboration with Severo Vazquez Prieto, Spain 
Organisation University of Santiago de Compostela
Department Department of Microbiology and Parasitology
Country Spain 
Sector Academic/University 
PI Contribution Expertise Intellectual known-how Resources - staff and consumables Sharing of data
Collaborator Contribution Collection of material Expertise Sharing of data and parasite material Resources - staff and consumables
Impact Manuscript in preparation
Start Year 2016
 
Description Hodgkinson-Brophy collaboration 
Organisation Aberystwyth University
Country United Kingdom 
Sector Academic/University 
PI Contribution 1. Advanced access to the Fasciola hepatica genome dataset provided in 2012. 2. Provision of biological material generated during this grant made in 2014 - specifically adult parasites of clonal Fasciola heptica isolates for collaboration of in vitro project looking at candidate gene approaches to triclabendazole resistance.
Collaborator Contribution Knowledge exchange between candidate triclabendazole resistance gene work currently underway in partner laboratories.
Impact Poster presentation at 52nd Spring Meeting of the British Society for Parasitology (BSP). 'Towards the Detoxome of Fasciola hepatica' Stuart, R. B. , Paterson, S Morphew, R. M. & Brophy, P. M.
Start Year 2014
 
Description Hodgkinson-Flynn collaboration 
Organisation University of Nottingham
Country United Kingdom 
Sector Academic/University 
PI Contribution 1. Advanced access to the Fasciola hepatica genome dataset. 2. Provision of novel biological material (generated during this grant) - specifically metacercariae of clonal Fasciola heptica isolates for gene specific studies.
Collaborator Contribution Knowledge exchange in the area of Fasciola heptica immune modulation.
Impact Manuscript entitled 'TGF-B superfamily members from the helminth Fasciola hepatica display parasite intrinsic effects on viability and development' O Japa, JE Hodgkinson, RD Emes RJ Flynn. Currently under review by International Journal for Parasitology
Start Year 2012
 
Description Hodgkinson-Maule collaboration 
Organisation Queen's University Belfast
Department Institute for Global Food Security
Country United Kingdom 
Sector Academic/University 
PI Contribution 1. Advanced access to the Fasciola hepatica genome dataset. 2. Provision of biological material generated during this grant - specifically metacercariae of clonal Fasciola heptica isolates for collaboration of in vitro project looking at candidate gene approaches to triclabendazole resistance. 3. Exchange of ideas around outputs and techniques available to study the biology of Fasciola hepatica e.g advanced knowledge of parasite population structure.
Collaborator Contribution 1. Training provided for postdoc to learn in vitro newly excysted juvenile (NEJ) assay to phenotype populations of Fasciola heptica for sensitivity to triclabendazole. 2. Access to expertise on the techniques available to study the biology of Fasciola hepatica e.g in vitro NEJ assay and RNAi.
Impact The nature of this collaboration is one of ongoing exchange of material and ideas around Fasciola hepatica biology and was the result of Professor Maule contacting me to present my work at his Institution in 2012. No immediate outputs are available but there are two manucripts currently in preparation with further collaborative work still underway. Most importantly this collaboration is ongoing and is mutually beneficial as we have complementary expertise that will move our work on in the longer term. For example, when candidiate genes for resistance are identified by our mapping experiments, Professor Maule has the expertise to test their role in resistance using his functional genomics platforms.
Start Year 2011
 
Description International Consortium for fluke genomics 
Organisation La Trobe University
Country Australia 
Sector Academic/University 
PI Contribution The Fasciola heptica genome was a resource in much demand from a number of International reseachers in the field. We were approached by a number of academic researchers in 2012 and put together an in formal 'Consortium for fluke genomics' via which I could coordinate access to the genome and biological material generated in the project. This served both to facilitate knowledge exchange, avoid duplication of scientific effort and as a source of future collboratation in the area of Fasciola spp biology. This led to a number of collaborations, specific details. Following publication of our first draft genome in 2015 we have deposited the assembly with Parasite wormbase.
Collaborator Contribution An early draft of the genome dataset (prior to publication or deposition in public databases) was made available to many consortium members listed here.
Impact A number of specfic collaborations resulted from this consortium, all of which are listed under seperate collaborations in this section.
Start Year 2012
 
Description International Consortium for fluke genomics 
Organisation Queen's University Belfast
Country United Kingdom 
Sector Academic/University 
PI Contribution The Fasciola heptica genome was a resource in much demand from a number of International reseachers in the field. We were approached by a number of academic researchers in 2012 and put together an in formal 'Consortium for fluke genomics' via which I could coordinate access to the genome and biological material generated in the project. This served both to facilitate knowledge exchange, avoid duplication of scientific effort and as a source of future collboratation in the area of Fasciola spp biology. This led to a number of collaborations, specific details. Following publication of our first draft genome in 2015 we have deposited the assembly with Parasite wormbase.
Collaborator Contribution An early draft of the genome dataset (prior to publication or deposition in public databases) was made available to many consortium members listed here.
Impact A number of specfic collaborations resulted from this consortium, all of which are listed under seperate collaborations in this section.
Start Year 2012
 
Description Paterson-Dalton collaboration 
Organisation Queen's University Belfast
Country United Kingdom 
Sector Academic/University 
PI Contribution Access to the Fasciola hepatica genome dataset to facilitate RNAseq profling of Fasciola hepatica life cycle stages.
Collaborator Contribution Acess to transcriptomic datasets from multiple life cycle stages to facilitate genome assembly and calling of gene models.
Impact Presentation of work as an oral presentation at the 24th World Association for the Advancement of Veterinary Parasitology (WAAVP), Perth, Australia, August 2013. Manuscript of the Fasciola heptica genome 'Gene duplication and polymorphism in the Fasciola hepatica genome reveals adaptation to the host environment and the capacity for rapid evolution.' K Cwiklinski, J P Dalton, P J. Dufresne, J La Course, D Williams, J Hodgkinson, S Paterson. Submitted to Genome Biology.
Start Year 2012
 
Description Arla and McDonalds 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Interest was sparked and substantial dicsussion afterwards.

Impact award to follow up control advice.
Year(s) Of Engagement Activity 2013,2014
 
Description Broughton Hall Outreach 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Institute of Infection and Global Health (IGH), University of Liverpool outreach program: a presentation regarding parasitology and careers in science at Broughton Hall High School, Liverpool.

Unknown impact as not directly recorded.
Year(s) Of Engagement Activity 2012
 
Description CPD 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact CPD: Member of Advisory team developing CPD for cattle and sheep vets on fluke control (supported by Sheep Veterinary Society and Industry). Took part in CPD workshop involving discussion of issues around fluke control.

CPD
Year(s) Of Engagement Activity 2013
 
Description Farming Connect (Wales) 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Presentation stimulated a lot of dicsussion amongst farmers and stake holders.

This led to follow-up testing of farms for problems of drug failure. This provided evidence with which to improve the understanding of other's in terms of the value of drug use on their farm. This allowed them to make more informed decisions about interventions and changed worming practice.
Year(s) Of Engagement Activity 2012
 
Description Fluke workshop held at University of Liverpool 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Policymakers/politicians
Results and Impact This workshop brought together leading academics, funders, industry, farmers, levy boards and producers to develop a strategic plan for the future fluke control including reducing spread of triclabendazole resistance.

The workshop provided evidence to feed into the guidelines document COWS (Control of Worms Sustainably) which inputs advice from leading academics, industry and levy boards to give advice to stakeholders (e.g farmers, producers). The COWS guidelines are available online.
Year(s) Of Engagement Activity 2013
URL http://www.cattleparasites.org.uk/
 
Description Germ Warcraft 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact July 2013 and November 2013: Germ Warcraft: two events run by IGH firstly at the Green Man Festival and secondly at the World Museum as part of the British Society of Immunology's Bug Busters Event. Children and their parents were taught about bacteria, viruses and parasites as well as the body's defences against them (antibody, macrophage, neutrophil).

Impact unknown as not formally captured.
Year(s) Of Engagement Activity 2013
 
Description IGH Food Safety Day 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact 15th June 2013: IGH Food Safety Day, this was an event that IGH organised in conjunction with the World Museum, Liverpool. We worked with other departments and the idea was to teach the general public about food safety, and we helped to organise activities from a parasitology point of view. Posters relating to the different parasites and their relation to food safety, as well as the fluke research we were conducting were created. We also organised activities including creating a giant model tapeworm, colouring sheets and pin the parasite on the part of the body it affects. Our activities were aimed at primary school children, although by them getting involved, we were also able to discuss our research with their parents.

Impact unknown as not formally captured.
Year(s) Of Engagement Activity 2013
 
Description Quality Meat Scotland; Edinburgh 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Keynote speaker at a day long workshop for farmers to advise of fluke control, which prompted much discussion afterwards.

Participation in subsequent BBSRC funded IPA project
Year(s) Of Engagement Activity 2013
 
Description The Art of Science Exhibition 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact 14th February 2014: The Art of Science Exhibition. This was an art exhibition organised by the University's School of Histories, Languages and Cultures. Scientific pictures were presented as an interactive display, alongside other scientists from within IGH. The pictures presented showed microscopic images of the reproductive cells of Fasciola, in some of the cells chromosomes were visible and the idea was for visitors to spot the chromosomes as well as learning a little about genetics and the impact that the parasite has. The exhibition was free and open to all and our pictures were displayed alongside more traditional artwork, therefore we were able to target a completely different demographic. The exhibition was particularly popular with young adults from a non-scientific background, and we even made cakes with our images printed on in icing.

Now that the exhibition has come to an end, the aim is to use this artwork within the Farm Animal areas at our Veterinary Field Station to help engage with farmers. Postcards with some of the pictures on were created with details of our research and funders.
Year(s) Of Engagement Activity 2014
URL http://www.liv.ac.uk/infection-and-global-health/news/stories/title,445228,en.html
 
Description The Cumbria Farmers' Network 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact The Cumbria Farmers' Network: investigating triclabendazole resistance in fluke populations in sheep in Cumbria. Presentations were made at two farmers' meetings in Sizergh and Penrith.

Resulted in press releases in local and regional papers and a series of sunbsequent meetings to British Cattle Veterinary Association.
Year(s) Of Engagement Activity 2013