Genetic and molecular basis of triclabendazole resistance in Fasciola hepatica
Lead Research Organisation:
University of Liverpool
Department Name: Institute of Infection and Global Health
Abstract
Fasciolosis is a common and important disease of livestock that results in substantial economic losses to the livestock sector. It is caused by the liver fluke, Fasciola hepatica, a flatworm found in the liver and transmitted via a mud snail. In the UK it is the most commonly reported infection associated with the digestive tract of ruminants. The immature stages of F. hepatica are responsible for acute disease, which has a negative impact on animal welfare and is often fatal in sheep. 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 pathogenic early immature stages. 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 reported in many locations worldwide. Little is known about the mechanisms of drug resistance in F. hepatica. This proposal aims to address this and other outstanding questions in drug resistance such as: Is there one genetic route to TCBZ resistance or multiple? What factors influence the emergence and spread of drug resistance alleles in liver fluke populations?
This project will utilise outputs from our highly successful previous project, which produced a draft F. hepatica genome and located (mapped) areas of the genome of TCBZ resistant parasites that may explain their ability to survive drug treatment. Here, we will produce a number of sequencing resources to improve this mapping exercise, including; a) a linkage map to improve the organisation of the genome and b) whole-genome sequencing of fluke eggs from the faeces of sheep naturally infected with resistant parasites, from before and after TCBZ treatment. Analysing these sequences will allow us to close in on the gene or genes responsible for encoding TCBZ resistance and will help us understand how resistance is passed from one parasite generation to the next. To assist in identifying the genes involved we will expose our resistant and susceptible clonal parasites to TCBZ and characterise the total gene expression profile and chemical fingerprint (global metabolomic profile). At the end of the project we will have genetic markers for resistance that can be used to detect and track the emergence of resistance in populations of F. hepatica naturally infecting sheep and cattle in the UK. There will be other important outputs for the wider scientific community from the project, such as a bank of genomic, transcriptomic and metabolomic data. We will make all these resources available to liver fluke researchers worldwide; so that, together, we can more effectively control this parasite in future.
This project will utilise outputs from our highly successful previous project, which produced a draft F. hepatica genome and located (mapped) areas of the genome of TCBZ resistant parasites that may explain their ability to survive drug treatment. Here, we will produce a number of sequencing resources to improve this mapping exercise, including; a) a linkage map to improve the organisation of the genome and b) whole-genome sequencing of fluke eggs from the faeces of sheep naturally infected with resistant parasites, from before and after TCBZ treatment. Analysing these sequences will allow us to close in on the gene or genes responsible for encoding TCBZ resistance and will help us understand how resistance is passed from one parasite generation to the next. To assist in identifying the genes involved we will expose our resistant and susceptible clonal parasites to TCBZ and characterise the total gene expression profile and chemical fingerprint (global metabolomic profile). At the end of the project we will have genetic markers for resistance that can be used to detect and track the emergence of resistance in populations of F. hepatica naturally infecting sheep and cattle in the UK. There will be other important outputs for the wider scientific community from the project, such as a bank of genomic, transcriptomic and metabolomic data. We will make all these resources available to liver fluke researchers worldwide; so that, together, we can more effectively control this parasite in future.
Technical Summary
The purpose of this project is to enhance our genetic resources for F. hepatica to support finer-scale mapping of triclabendazole (TCBZ) resistance loci. We will integrate the mapping study with in vitro and in vivo transcriptomic and global metabolomic profiling of TCBZ-resistant and -susceptible clonal isolates to generate a defined set of candidate genes for TCBZ resistance. The project is divided into four related objectives:
Objective 1. Deliver a genetic map for Fasciola hepatica and determine if TCBZ-R is a dominant or recessive trait.
Objective 2. Validate putative TCBZ resistance markers and perform finer-scale mapping of TCBZ resistance loci using field isolates of known sensitivity to TCBZ.
Objective 3. Compare transcript abundance in TCBZ-resistant (TCBZ-R) and TCBZ-susceptible (TCBZ-S) parental parasites with and without exposure to TCBZ in vivo and in vitro by RNAseq.
Objective 4. Determine metabolomic differences between TCBZ-R and TCBZ-S parasites with and without exposure to TCBZ in vivo and in vitro.
The project aims to determine whether TCBZ resistance has a common genetic basis across the UK, whether the same mechanism(s) is employed by both adult and immature parasites and whether TCBZ resistance is a dominant or recessive trait. Detecting TCBZ resistance alleles in the field will provide insights into practices influencing development of resistance and ultimately mitigate against the spread of drug resistance.
Objective 1. Deliver a genetic map for Fasciola hepatica and determine if TCBZ-R is a dominant or recessive trait.
Objective 2. Validate putative TCBZ resistance markers and perform finer-scale mapping of TCBZ resistance loci using field isolates of known sensitivity to TCBZ.
Objective 3. Compare transcript abundance in TCBZ-resistant (TCBZ-R) and TCBZ-susceptible (TCBZ-S) parental parasites with and without exposure to TCBZ in vivo and in vitro by RNAseq.
Objective 4. Determine metabolomic differences between TCBZ-R and TCBZ-S parasites with and without exposure to TCBZ in vivo and in vitro.
The project aims to determine whether TCBZ resistance has a common genetic basis across the UK, whether the same mechanism(s) is employed by both adult and immature parasites and whether TCBZ resistance is a dominant or recessive trait. Detecting TCBZ resistance alleles in the field will provide insights into practices influencing development of resistance and ultimately mitigate against the spread of drug resistance.
Planned Impact
This project will maximise outputs from existing liver fluke research projects, stimulate new areas of research and facilitate translation of research to farmers and industry. There are a number of beneficiaries from this proposal.
Academic impact
This project will deliver an enhanced genome assembly and a linkage map for F. hepatica; vital to mapping complex genetic traits in future. The datasets we generate will prove invaluable to the F. hepatica research community. They will build on the limited transcriptomic resources available to the liver fluke research community and deliver the first global metabolomic datasets. Our particular focus on NEJ, the primary target of control strategies, will generate transcriptional and metabolomic datasets for the development of new anthelmintics, diagnostics and vaccines. One of the most important outputs from the project will be post-doctoral scientists trained in the most up to date methodologies including bio-informatics and analysis of genomic, transcriptome and metabolome data. This will contribute to a cadre of highly skilled biomedical scientists to build science capacity in the UK.
Impact for UK agriculture
Agriculture is worth around £11 billion to the UK economy. Securing the UK's food supply through efficient production of beef, lamb and dairy products is essential for the health and well being of the UK population. Moreover efficient production systems will reduce the emission of greenhouse gasses from livestock. Liver fluke is a substantial constraint on meat and milk production in the UK. The parasite affects feed conversion, growth rates and milk yield and has a negative impact on farmers' incomes and climate change is predicted to exacerbate fasciolosis throughout this century. Control of fluke is now impeded by resistance to triclabendazole and this represents a major threat, particularly to sheep production in marginal areas such as upland Wales and Cumbria. By identifying the genetic regions involved in TCBZ resistance and determining whether TCBZ resistance has the same genetic basis at different UK locations, markers for TCBZ resistance can be developed that will be used to detect drug resistance alleles in the field. This in turn will provide insights into practices influencing the emergence and spread of drug resistance in F. hepatica populations, directly benefit farming communities and the agri-food sector by reducing parasite burdens and improving productivity and biological efficiency.
Stakeholders and Policy impact
This proposal will deliver an evidence-base for how to combat the development and spread of drug resistance which will feed into policy development by UK-based government bodies such as the Dept. for Environment, Food & Rural Affairs (DEFRA), the agriculture and horticultural development boards (AHDB) and APHA as well as the Department of Agriculture and Rural Development-NI. Through our links with the Control of Worms Sustainably industry stakeholder group (Co-I Williams), we have a direct route to promote best practice in parasite control.
Wider public
Consumers are becoming increasingly aware of the quality and provenance of food and it is increasingly important to the wider public that they have a reliable source of affordable and healthy food, without compromising animal health and welfare. Beef and dairy product supplies, their quality, environmental impact, price and welfare of the animals from which they are produced will be affected by the prevalence of fasciolosis. Public engagement and outreach activities are embedded within the Institute of Infection and Global Health, UofL. We will interact with farmers and the public through events such as LEAF Open Farm Sunday.
Fasciolosis is listed amongst the neglected tropical diseases affecting people in resource poor countries. Improved means of diagnosing and treating liver fluke infection will be of substantial benefit to people in these settings.
Academic impact
This project will deliver an enhanced genome assembly and a linkage map for F. hepatica; vital to mapping complex genetic traits in future. The datasets we generate will prove invaluable to the F. hepatica research community. They will build on the limited transcriptomic resources available to the liver fluke research community and deliver the first global metabolomic datasets. Our particular focus on NEJ, the primary target of control strategies, will generate transcriptional and metabolomic datasets for the development of new anthelmintics, diagnostics and vaccines. One of the most important outputs from the project will be post-doctoral scientists trained in the most up to date methodologies including bio-informatics and analysis of genomic, transcriptome and metabolome data. This will contribute to a cadre of highly skilled biomedical scientists to build science capacity in the UK.
Impact for UK agriculture
Agriculture is worth around £11 billion to the UK economy. Securing the UK's food supply through efficient production of beef, lamb and dairy products is essential for the health and well being of the UK population. Moreover efficient production systems will reduce the emission of greenhouse gasses from livestock. Liver fluke is a substantial constraint on meat and milk production in the UK. The parasite affects feed conversion, growth rates and milk yield and has a negative impact on farmers' incomes and climate change is predicted to exacerbate fasciolosis throughout this century. Control of fluke is now impeded by resistance to triclabendazole and this represents a major threat, particularly to sheep production in marginal areas such as upland Wales and Cumbria. By identifying the genetic regions involved in TCBZ resistance and determining whether TCBZ resistance has the same genetic basis at different UK locations, markers for TCBZ resistance can be developed that will be used to detect drug resistance alleles in the field. This in turn will provide insights into practices influencing the emergence and spread of drug resistance in F. hepatica populations, directly benefit farming communities and the agri-food sector by reducing parasite burdens and improving productivity and biological efficiency.
Stakeholders and Policy impact
This proposal will deliver an evidence-base for how to combat the development and spread of drug resistance which will feed into policy development by UK-based government bodies such as the Dept. for Environment, Food & Rural Affairs (DEFRA), the agriculture and horticultural development boards (AHDB) and APHA as well as the Department of Agriculture and Rural Development-NI. Through our links with the Control of Worms Sustainably industry stakeholder group (Co-I Williams), we have a direct route to promote best practice in parasite control.
Wider public
Consumers are becoming increasingly aware of the quality and provenance of food and it is increasingly important to the wider public that they have a reliable source of affordable and healthy food, without compromising animal health and welfare. Beef and dairy product supplies, their quality, environmental impact, price and welfare of the animals from which they are produced will be affected by the prevalence of fasciolosis. Public engagement and outreach activities are embedded within the Institute of Infection and Global Health, UofL. We will interact with farmers and the public through events such as LEAF Open Farm Sunday.
Fasciolosis is listed amongst the neglected tropical diseases affecting people in resource poor countries. Improved means of diagnosing and treating liver fluke infection will be of substantial benefit to people in these settings.
Organisations
Publications
Beesley NJ
(2023)
A major locus confers triclabendazole resistance in Fasciola hepatica and shows dominant inheritance.
in PLoS pathogens
Beesley NJ
(2021)
Evidence of population structuring following population genetic analyses of Fasciola hepatica from Argentina.
in International journal for parasitology
Beesley NJ
(2018)
Fasciola and fasciolosis in ruminants in Europe: Identifying research needs.
in Transboundary and emerging diseases
Beesley NJ
(2017)
Fasciola hepatica demonstrates high levels of genetic diversity, a lack of population structure and high gene flow: possible implications for drug resistance.
in International journal for parasitology
Hodgkinson JE
(2019)
Refugia and anthelmintic resistance: Concepts and challenges.
in International journal for parasitology. Drugs and drug resistance
John BC
(2020)
Anaerobic fermentation results in loss of viability of Fasciola hepatica metacercariae in grass silage.
in Veterinary parasitology
John BC
(2019)
A review of our current understanding of parasite survival in silage and stored forages, with a focus on Fasciola hepatica metacercariae.
in Grass and forage science : the journal of the British Grassland Society
Kamaludeen J
(2019)
Lack of efficacy of triclabendazole against Fasciola hepatica is present on sheep farms in three regions of England, and Wales.
in The Veterinary record
| Description | Liver fluke parasites impact on the health and welfare of sheep and cattle in the UK and globally. In some areas of the world, for example South America, the liver fluke causes disease in humans, and it is recognised by WHO as a neglected tropical disease. Only a few drugs can treat infection with liver fluke, one such drug is triclabendazole; the drug of choice to treat livestock and the only drug licenced to treat humans. However, liver fluke parasites are becoming resistant to triclabendazole, which is impacting negatively on our ability to effectively control and treat infection with this parasite. This project set out to identify what genetic changes occur in triclabendazole resistant parasites and how resistance is inherited from one parasite generation to another. We were also interested to see if there is just one route to resistance or if different mechanisms of resistance occur in different parasite populations infecting UK livestock. Our most significant achievements include: 1. An enhanced genome assembly for the liver fluke, Fasciola hepatica. 2. We have identified that triclabendazole resistance is caused by gene(s) that occupy a single region (locus) within the genome. 3. We have determined that one of 15 genes is responsible for conferring triclabendazole resistance and we have shown that drug resistant F. hepatica show lower levels of gene expression in both juvenile and adult parasites. 4. We used an experimental system to identify triclabendazole resistance genes, but we have shown that this same region of the genome is involved in conferring resistance in naturally infected sheep following treatment with triclabendazole on farm. 5. We have demonstrated that triclabendazole resistance shows dominant inheritance, meaning that just one resistance allele needs to be passed on to future generations for them to survive treatment. 6. This dominant inheritance combined with our knowledge of genes in parasite populations tells us that resistance can spread rapidly and emphasises the importance of quarantine treatments for farmers to prevent introducing resistance onto their farm. Our work has addressed all the objectives laid out in the initial proposal. We have shown how drug resistance is inherited and that a common mechanism of resistance is found in liver fluke populations from different farms in the UK. Our functional studies have demonstrated how both juvenile and adult drug-resistant and -susceptible parasites respond to drug exposure. We have narrowed down the causal gene for triclabendazole resistance to one of 15 genes, paving the way to development of a molecular test to detect resistance that can be used in the UK and in the many areas of the world where F. hepatica is endemic. |
| Exploitation Route | Amongst the beneficiaries from the research outcomes are industry and commercial enterprises. Liver fluke is a substantial threat to animal welfare, productivity and profitability by the agricultural industry. Fasciolosis is a significant constraint on beef and milk production in the UK and has been estimated to cost the UK livestock industry £300 million per year. The parasite affects feed conversion, growth rates and milk yield and has a negative impact on farmers' incomes. Over the last decade liver fluke infection in the UK has increased and infection has extended into new areas, in part due to warmer, wetter winters. Resistance of liver fluke to drugs, particularly triclabendazole poses a significant threat to the future treatment of fasciolosis and control of liver fluke infection, especially in sheep. During the course of this project we have worked with multiple farming enterprises to detect F. hepatica infection on farm and established to what extent triclabendazole treatment is an effective means of control. This work has been highlighted by industry bodies such as Agriculture and Horticultural Development Board (AHDB) and has fed into initiatives such as Control of Worms Sustainably. Our finding that resistance is a dominant trait highlights that once introduced on farm resistance has the potential to spread rapidly, emphasising the need for good biosecurity and quarantine procedures when introducing new livestock on farm. The project has delivered a foundation for a molecular diagnostic to detect triclabendazole resistance in an important livestock pathogen, this can now be taken forward to functional studies to determine the cause resistance in liver flukes and ultimately it can be developed as a diagnostic to track the emergence and spread of triclabendazole resistance alleles in F. hepatica; this will directly benefit farming communities and the agri-food sector and improve productivity and biological efficiency. Beyond the immediate need for a diagnostic for resistance the outcome of a publicly available and enhanced draft F. hepatica genome has provided a global resource that has fuelled many academic enterprises, including vaccine research, functional biology studies and the identification of novel drug targets within the parasite. As a resource the genome has underpinned multiple PhD student projects and research opportunities. It is a fundamental resource to the academic parasitology community and will continue to provide a platform for research. |
| Sectors | Agriculture, Food and Drink,Education,Pharmaceuticals and Medical Biotechnology |
| URL | https://parasite.wormbase.org/Fasciola_hepatica_prjeb25283/Info/Index/ |
| Description | This work has been used to improve the effectiveness of public services and policy. Within the veterinary sector it has raised the profile of drug (triclabendazole) resistance in liver fluke populations and driven a greater emphasis on testing livestock for infection prior to and post treatment as a means of improving livestock health, welfare and production efficiency. This project has raised the profile of the problem of triclabendazole specifically but has formed part of the wider debate about the need for reduced anthelmintic use on farm to slow down resistance development. This has fed through to policy making, particularly in regard to a move towards greater accountability in the prescription of POM-VPS products and the point of sale prescription process. We have worked with sheep farmers in the UK who are very concerned about losing their livestock to fasciolosis. By working with them to identify if triclabendazole is effective in their livestock and detecting those who have problems with resistance, we have been able to make informed decisions about the use of alternative, effective anthelmintics. This has meant that farmers have been able to better control the parasite in their livestock and led to improved outcomes for all. Within the academic community there has been substantial interest in the mechanism of triclabendazole resistance over the last two decades, but it has remained elusive. Our findings have provided the first definitive evidence that triclabendazole is a genetic trait and narrowed down the causal gene to one of four primary candidates. Our genetic crossing experiment in vivo has paved the way for determining further genetic traits in Fasciola hepatica in future. |
| First Year Of Impact | 2019 |
| Sector | Agriculture, Food and Drink,Education,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Societal,Economic,Policy & public services |
| Description | BBSRC DTP SCHEME UNIVERSITIES OF LIVERPOOL, DURHAM AND NEWCASTLE |
| Amount | £80,000 (GBP) |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 10/2019 |
| End | 09/2023 |
| 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:Research Grant United States-UK partnering award |
| Amount | £49,898 (GBP) |
| Funding ID | BB/N022386/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 08/2016 |
| End | 07/2021 |
| 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 | 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/ |
| Title | FhLiv enhanced genome |
| Description | An enhanced genome has been generated following additional sequencing using Dovetail and 10x platforms. This has significantly increased the N50 value and allowed more efficient mapping of genetic traits e.g drug resistance genes |
| Type Of Material | Database/Collection of data |
| Year Produced | 2018 |
| Provided To Others? | Yes |
| Impact | Newer. improved version of annotated genome provides a high quality reference genome for the wider liver fluke reserach community. |
| URL | https://parasite.wormbase.org/Fasciola_hepatica_prjeb25283/Info/Index/ |
| Title | Metabolomic profile of liver fluke and isolate to isolate comparison |
| Description | This is the first global metabolomic profile of adult liver fluke parasites and it provides a comparison of metabolites produced by clonal isolates in response to drug exposure. |
| Type Of Material | Database/Collection of data |
| Year Produced | 2020 |
| Provided To Others? | No |
| Impact | This analysis will provide a very valubale reference dataset for metabolomic studies on this parasite |
| Title | RNAseq dataset from clonal isolates with and without drug exposure |
| Description | We have performed in vivo infection of sheep with six clonal isolates and each clone was either exposed to drug or left untreated. This dataset is a unique resource of isolates and RNAseq data from drug susceptible and -resistant parasites |
| Type Of Material | Database/Collection of data |
| Year Produced | 2020 |
| Provided To Others? | No |
| Impact | We are just now analysing these data but it will provide answers to both the mode of action of the drug and the mode of drug resistance. |
| Description | Collaboration with Aaron Maule, Queen's University Belfast |
| Organisation | Queen's University Belfast |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We are collaborating in a number of ways: 1. To better define our clonal liver fluke isolates - I own the isolates, grow and maintain them. 2. To expose NEJ to triclabendazole and its metabolites in vitro for metabolomic and RNAseq analysis - we are running the assays and performing the metabolomics and RNAseq |
| Collaborator Contribution | We are collaborating in a number of ways: 1. To better define our clonal liver fluke isolates - quantitate neoblasts in each isolate 2. To expose NEJ to triclabendazole and its metabolites in vitro for metabolomic and RNAseq analysis - ran in vitro assays to generate LD50 values 3. Knowledge transfer of immature parasite (NEJ) culture in vitro for RNAseq and metabolomic expts and for future RNAi knockdown |
| Impact | Exposing a role for liver fluke stem cells in the flukicide response. Nathan Clarke; Duncan Wells; Erica Gardiner; Paul McCusker; Emily Robb; Paul McVeigh; Jonathan Coulter; Jane Hodgkinson; Nikki J. Marks; Aaron G. Maule; The Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast; School of Pharmacy, Queen's University Belfast; Institute of Global Health, University of Liverpool, Liverpool, UK, |
| Start Year | 2016 |
| Description | Human fasciolosis with Dr Nick Beeching |
| Organisation | Liverpool School of Tropical Medicine |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Collaboration on case of human fasciolosis leading to surgical removal of parasites and transfer of material to our laboratory for genotyping. Presentation at two medical tropical diseases case presentations. |
| Collaborator Contribution | Provision of Fasciola spp from human infection |
| Impact | Genotyping of flukes is ongoing |
| Start Year | 2021 |
| Description | Russian collaboration |
| Organisation | Russian Academy of Sciences |
| Country | Russian Federation |
| Sector | Public |
| PI Contribution | Transposable elements in Fasciola hepatica with Dr. Anna Solovyeva and Prof. Olga podgornaya, Russian Academy of Sciences As the parasitologists we performed triplicate in vitro culture of our parasite to generate RNE and then cDNA of F.hepatica from the following life-cycle stages: eggs metacercariae 1 hr newly excysted juvenile 3 hr newly excysted juvenile 24hr newly excysted juvenile 21day juvenile We also generated cDNA from adult material from our archive. |
| Collaborator Contribution | Dr. Anna Solovyeva and Prof. Olga podgornaya, Laboratory of Non-coding DNA, Russian Academy of Sciences, Institute of Cytology They performed some in silico research with our published data and revealed that transposable elements have various ranges of transcription at different stages of the F.hepatica lifecycle. Since transposons have many ways of influencing the genome, they wanted to understand if they could play a role, necessary for trematode lifecycle regulation. They approached us by email in July 2020 to collaborate together in this work. |
| Impact | Outputs not yet realised but at least one manuscript is being drafted. We were in the process of drafting the manuscript but in light of the illegal invasion of Ukraine by Russia and following the instruction of my institution I have terminated the collaboration with immediate effect. |
| Start Year | 2020 |
| Description | University of Liverpool-Freie Universität Berlin |
| Organisation | Free University of Berlin |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | Propagation of a German triclabendazole resistant isolate through our snail colony so that the isolate can be maintained and studies. Sharing of protocols for the processing of fluke material and assessing drug efficacy. |
| Collaborator Contribution | Sharing of parasite material (eggs of Fasciola). |
| Impact | New collaboration - one publication currently under review. |
| Start Year | 2022 |
| Description | COWS meeting |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Public/other audiences |
| Results and Impact | COWS Steering Group Meeting followed by talk to approx. To farmers / agricultural students on liver fluke at Bridgwater College, Somerset, UK |
| Year(s) Of Engagement Activity | 2017 |
| Description | Grant workshop for early career researchers |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | To provide support for early career researchers within the University of Liverpool Faculty of Health and Life Sciences. Presented by BBSRC and other UKRI-funded PDRAs to their peers and postgraduate students. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Invited Keynote Australian Society for Parasitology |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Elsevier invited speaker IJP: Drugs and Drug Resistance Lecturer at the 2020 Australian Society for Parasitology Annual Conference in Cairns, Australian. July 2022. There was much interest in my presentation and it prompted an online discussion. It would have been nice to attend in person, but it was not possible on this occasion. The conference had been postponed due to Covid and was run in hybrid format. I was subsequently approached by an academic who was interested in collaborating in future. |
| Year(s) Of Engagement Activity | 2022 |
| Description | Keynote presentation at International Conference for Parasitology [ICOPA) |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | ICOPA is the most prestigious and largest conference hosted within the international parasitology community. It has >1000 delegates and highlights all areas of human, veterinary and zoonotic parasitology. I was delighted to be invited to talk about our work on determining the genetic basis of anthelmintic resistance in a common liver fluke, Fasciola hepatica. My keynote presentation was focused on work which has significantly advanced our understanding of how this parasite becomes resistant to the most efficacious flukicide, triclabendazole and highlighted our substantial progress in identifying the gene(s) conferring resistance. This work was entirely funded by BBSRC and has generated a number of vital resource for the liver fluke community, namely the first liver fluke genome draft and clonal liver fluke isolates of known genotype. Fasciola spp infections impact on both livestock and human health in many regions of the world, their was much discussion after my talk about what this might mean for detection of resistance in many host species and deployment of efficacious drugs to prevent disease due to this parasite. After my presentation I was approached by several academics at all career levels who were interested in future collaborations. |
| Year(s) Of Engagement Activity | 2022 |
| Description | Keynote presentation: BSP British Society for Parasitology Autumn symposium 'Molecular basis of triclabendazole resistance in Fasciola hepatica field isolates' Queens University Belfast. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Keynote presentation: BSP British Society for Parasitology Autumn symposium 'Molecular basis of triclabendazole resistance in Fasciola hepatica field isolates' Queens University Belfast. To provide a holistic overview of the project aims and objectives and our outputs so far. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Liverpool Metabolomics Symposium: Oral presentation: Optimising extraction of liver fluke: considerations for host-parasite metabolom |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Professional Practitioners |
| Results and Impact | Conference presentation to highlight approaches taken to optimise novel metabolomics of liver fluke. It provided a benchmark for others to follow. |
| Year(s) Of Engagement Activity | 2019 |
| Description | Newton Research Links Bilateral Workshop: A showcase of the Liver fluke research @ Liverpool with specifics on the changes in metabolites following exposure to triclabendazole in triclabendazole-resistant and triclabendazole-susceptible liver fluke |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Competitive application to Newton fund for postdoctoral researcher, Nicola Beesley, to attend a workshop for early career researchers, in China, December 2019 |
| Year(s) Of Engagement Activity | 2019 |
| Description | Oral presentation at Anti-helminthics Drugs, Resistance and Vaccines, UMass, Worcester, MA, USA |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | This meeting is delivered as a small, focused meeting that can bring together the international anti-helminthics community for talks and discussions. I gave a short presentation to inform the audience about how are drug-susceptible and -resistant parasites respond differently on drug exposure in vivo. This work was novel and is currently being written up for publication. This presentation allowed for critique and discussion within a knowledgeable community which will prove useful for future studies. I took two PhD students out to the meeting (their first chance to go to an international meeting post covid) and there was good representation of PGR students, which gave them a great opportunity to meet their international peers. A new collaboration with a group in Argentina was forged. |
| Year(s) Of Engagement Activity | 2022 |
| Description | Panning for Parasites |
| 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 | 08.07.17: Meet the Scientists at the World Museum, Liverpool - Panning for Parasites - activity teaching children about faecal egg counts - including averages, interpreting negative results etc. engaged with approx. 150 children and their parents. |
| Year(s) Of Engagement Activity | 2017 |
| Description | Presentation at National Conference - British Society for Parasitology |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Invited presentation to promote BBSRC-funded work on drug resistance within liver fluke. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Python training scheme at UoL |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Schools |
| Results and Impact | 16.06.17: Public engagement activity with HiPy (Python training scheme at UoL) widening participation schools - teaching Python coding using Codecademy and explaining how we use coding in science. 2 groups of approx. 20 students |
| Year(s) Of Engagement Activity | 2017 |
| Description | Scientific presentation |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | 23.05.17: Around 70 vets and industry stakeholders plus a few farmers attended the Sheep Veterinary Society, Cornwall, 2016. A presentation was given to explain current status of triclbendazole resistance on farm following which there was an active debate about current practice of parasite control with drugs and the problems farners are encountering. It was a very useful two way process of knowledge exchange. |
| Year(s) Of Engagement Activity | 2016 |
| Description | Twitter account |
| Form Of Engagement Activity | Engagement focused website, blog or social media channel |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | 15.01.18: Set up @flukeresistance twitter - now with 137 followers |
| Year(s) Of Engagement Activity | 2017 |
| Description | Veterinary Alumni Celebration: Outreach presentation to veterinary graduates from University of Liverpool ranging from 1960 to 2019 graduates. Title of talk: The 'omics revolution: an example from veterinary parasitology |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Nicola Beesley, PDRA on this project presented the project aims and objectives with a focus on the training it had provided for her during her appointment. |
| Year(s) Of Engagement Activity | 2020 |
| Description | WAAVP 2019 Madison Wisconsin: Oral presentation: Does triclabendazole resistance in Fasciola hepatica have the same genetic basis in different areas of the UK |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Conference presentation highlighting drug resistance development and spread |
| Year(s) Of Engagement Activity | 2019 |