Knowledge Transfer -Impacts of inbreeding on the responses to pharmaceuticals and fitness consequences in fish - a molecular approach
Lead Research Organisation:
UNIVERSITY OF EXETER
Department Name: Biosciences
Abstract
Pharmaceuticals in the environment potentially have wide reaching deleterious impacts on wildlife and possibly humans, and are at the top of the Environment Agency's agenda for environmental pollution. Population level declines in wildlife have resulted from exposure to pharmaceuticals (e.g. diclofenac in vultures in Asia). Furthermore, some reproductive disorders associated in humans are now a major health issue and parallel those effects seen in wildlife that have been associated with exposure to environmental chemicals, including pharmaceuticals. The UK holds a strong lead in the field of pharmaceuticals in the aquatic environment and the applicants have contributed to the unfolding explanation of how some such contaminants interfere with normal reproductive activity in fish. . Information on interactive effects of inbreeding and pollutant exposure is extremely limited and essentially nothing is known for pharmaceuticals, which is surprising given that inbred strains of animals are used most commonly for laboratory testing of chemicals, including pharmaceuticals. This project will provide an understanding of the vulnerability or resistance of inbred populations of fish for exposure to the pharmaceutical,ketoconazole (a synthetic antifungal drug employed as an agricultural pesticides and clinically to prevent and treat skin and fungal infections), as a representative of a group of widely used group of fungicides. The use of genome-wide approaches (gene arrays) will enable us to dissect out gene expression networks most intimately associated with physiological stress for exposure to this pharmaceutical. The data from the project will further provide insights into the critical determinants of vulnerability of fish to a pharmaceutical of importance to human health. The data on the comparative responses to ketoconazole in inbred and outbred populations and impacts on breeding dynamics will also be highly informative for ecological and environmental risk assessment. This proposal will thus provide new information on gene-environment interactions for chemical exposure effects in fish. The project is also likely to deliver a more informative set of biological (molecular) effects tools upon which to base environmental impact and health assessments for ketoconazole and other allied fungicides. This work will provide substantive information on the health impacts of ketoconazole in the natural environment for the better protection of our aquatic resources and biodiversity and will contribute more widely to environmental impact assessments and will thus be of very wide interest to the government regulatory bodies, environment protection groups, industry and the wider public. A key element of the project is an enhancement in the partnership between academia (University of Exeter) and an end-user (Astrazeneca). This partnership will facilitate the uptake of genetic and genomic methods for chemical hazard identification (and developed through NERC funded science) into the pharmaceutical industry. It will further facilitate an enhanced understanding in academia of the practical research needs of the chemical/pharmaceutical industry.
People |
ORCID iD |
Charles Tyler (Principal Investigator) | |
David Hosken (Co-Investigator) |
Publications
Bickley LK
(2009)
Evaluation of a carp primary hepatocyte culture system for screening chemicals for oestrogenic activity.
in Aquatic toxicology (Amsterdam, Netherlands)
Bickley LK
(2017)
Bioavailability and Kidney Responses to Diclofenac in the Fathead Minnow (Pimephales promelas).
in Environmental science & technology
Bickley LK
(2013)
Interactive effects of inbreeding and endocrine disruption on reproduction in a model laboratory fish.
in Evolutionary applications
Brown AR
(2011)
Are toxicological responses in laboratory (inbred) zebrafish representative of those in outbred (wild) populations? - A case study with an endocrine disrupting chemical.
in Environmental science & technology
Brown AR
(2009)
Genetic variation, inbreeding and chemical exposure--combined effects in wildlife and critical considerations for ecotoxicology.
in Philosophical transactions of the Royal Society of London. Series B, Biological sciences
Brown AR
(2015)
Climate change and pollution speed declines in zebrafish populations.
in Proceedings of the National Academy of Sciences of the United States of America
Brown AR
(2012)
Differences in sexual development in inbred and outbred zebrafish (Danio rerio) and implications for chemical testing.
in Aquatic toxicology (Amsterdam, Netherlands)
Corcoran J
(2010)
Pharmaceuticals in the aquatic environment: a critical review of the evidence for health effects in fish.
in Critical reviews in toxicology
Hogstrand, Christer; Kille, Pete
(2008)
Comparative Toxicogenomics
Lange A
(2017)
Development of a common carp (Cyprinus carpio) pregnane X receptor (cPXR) transactivation reporter assay and its activation by azole fungicides and pharmaceutical chemicals.
in Toxicology in vitro : an international journal published in association with BIBRA
Description | What have you discovered or developed through the research funded on this grant? Please explain for a non-specialist audience. We undertook to collect zebrafish from wild populations in Bangladesh to investigate whether their responses to chemical challenge were comparable with zebrafish that have been maintained in the laboratory for many generations (and were inbred) that are used widely for standardised OECD tests. The studies n included making assessment on the interactive effects of the fish genetics (inbred versus outbred lines of zebrafish) and an endocrine disrupting chemical (clotrimazole) on sexual development and breeding. Clotrimazole affects an enzyme called aromatase that is involved with the conversion of testosterone to oestrogen and is thus fundamental in the process of sexual differentiation in fish (reinforcing the feminised phenotype). We show that there were differences in responses (sensitivity for effect) when considering the genetics of the zebrafish for chemical exposure and this has implications when considering the use of zebrafish lines in standardised regulatory chemical testing. Furthermore, the research identified that the timings differ for gonadal sexual differentiation for inbred versus outbred zebrafish with direct implications for use of an established OECD test guideline - where gonadal sex is a key endpoint for chemical effects assessments. The research undertaken highlights the need to better reporting on the genetics of fish strains/lines used for chemical testing (and indeed research more generally) that would more accurate comparisons for research findings between different laboratories adopting different fish strains/lines. The research undertaken in this project has been disseminated widely through conferences and workshops to relevant stakeholders and been featured in three full primary data research papers: Brown AR, Bickley LK, Le Page G, Hosken D, Paull,GC , Hamilton PB, Owen SF, Robinson,J., Sharpe AD and Tyler CR* (2011). Are toxicological responses in laboratory (inbred) zebrafish representative of those in outbred (wild) populations? - a case study with an endocrine disrupting chemical. Environmental Science and Technology 45(9):4166-4172 A Ross Brown , Lisa K Bickley, Thomas Ryan, Greg C Paull , Patrick B Hamilton, Stewart F Owen, Alan D Sharpe and Charles R Tyler* (2012). Differences in sexual development in inbred and outbred zebrafish (Danio rerio) and implications for chemical testing. Aquatic Toxicology 112-113, 27-38. Bickley LK, Brown AR, Le Page G, Hosken D, Paull,GC , Hamilton PB, Owen SF, Wilga J, Hetheridge MJ, Sharpe AD and Tyler CR (2012). Interactive effects of inbreeding and endocrine disruption on reproduction in a model laboratory fish. Evolutionary Applications. 6: 279-289 The research undertaken has also resulted in the production of two review paper papers to further highlight some of the critical issues identified during this research: Brown, R.,Hosken,D., Balloux, F., Bickley, L.L., LePage, G. Owen S., Hetheridge M Tyler, C.R.* (2009). Genetic variation, inbreeding and chemical exposure - combined effects in wildlife and critical considerations for ecotoxicology. Phil Trans Royal Soc. 364, 3377-3390 Brown A R, Gunnarsson L M, Kristiansson E, Tyler C R (2014). Evolutionary, Ecological, and Life History Considerations in the Variation of Susceptibility to Pharmaceuticals in Fish. Phil Trans Royal Soc. 369 1656 - 20130576. This work has furthermore lead to an analysis (ongoing) that is addressing the possible implications for fish populations of exposure to clotrimazole, together with changes in climate (temperature elevation) and inbreeding depression. |
Exploitation Route | see impact |
Sectors | Chemicals,Communities and Social Services/Policy,Education,Environment,Government, Democracy and Justice,Pharmaceuticals and Medical Biotechnology |
Description | A major impact of this research project was the knowledge transfer to Industry (AstraZeneca) from Exeter on the use and application of a wide range of molecular tools and techniques, including DNA extraction techniques, q RT-PCR, and DNA microsatellites for their application in (eco)toxicology. Equally from this partnership Exeter gained an enhanced understanding of end user needs for the chemical/pharmaceutical industry, and access to world class chemical exposure and analysis facilities. Exchanges of researchers (Ross Brown and Lisa Bickley) provided the main conduit for transfer of information between the collaborating laboratories. Reciprocal placements enabled these researchers to gain complimentary skills sets and understanding of the different (academic vs industry) environments. Ross Brown subsequently registers (and duly obtained) his PhD from Exeter. Wider links were gained for research and exchange of personnel through this research grant, leading to further grants, PhD studentship and training. This grant was important in forging an AstraZeneca -University of Exeter partnership that secured a DTi Knowledge Transfer Project to develop further the application of molecular tools in ecotox testing and significantly, recently (2014) resulted in a £1.3M AZ funded research and development project to Exeter. This project produced zebrafish lines with improved genetic variation that were supplied to AstraZeneca for their chemical testing programmes. In the collaborative research project these lines were used investigate to what extent chemical exposures in inbred fish (as held for most testing labs) can be used to inform on effects for wild ( out-bred fish), with significant findings and questions for standardised chemical testing. This work from this project has resulted in a series of high impact research papers, international conferences presentations and further research partnerships. The results of this work made a significant impact on the general public and environmental community, with Brown et al., 2015. PNAS doi: 10.1073/pnas.1416269112 prompting several cover stories and editorials in 2015 (reported on previously) and subsequently in 2016: • Atlas of Science (14 March 2016). https://atlasofscience.org/climate-change-may-add-significantly-to-the-impacts-of-pollution-and-habitat-loss-on-fish-populations/ Online impact of PNAS paper: Altmetric Score: 39 Total downloads: Abstract = 9585 Manuscript = 1483 PDF = 6972 (Article received 5704 hits on the abstract and 1000 downloads in the first month) We have informed the UK (e.g. Environment Agency and DEFRA), US and Japanese governments, European Union, environment protection groups, Industry and the wider public (e.g. via the Royal Society, Zoological Society London, local conservations groups) on this work. Information has been exchanged further through is existing NERC Knowledge Transfer Networks on contaminants in the aquatic environment that involve conferences and workshops which are attended by academics, regulators and industry as well as experienced researchers and research students. We have also been active in disseminating our research work to Schools, and higher education programmes, principally through University Open Days and International University seminars. |
First Year Of Impact | 2015 |
Sector | Chemicals,Communities and Social Services/Policy,Education,Environment,Government, Democracy and Justice,Pharmaceuticals and Medical Biotechnology |
Impact Types | Cultural,Societal,Economic |
Description | AstraZeneca - £1.3m |
Amount | £1,300,000 (GBP) |
Organisation | AstraZeneca |
Sector | Private |
Country | United Kingdom |
Start | |
End | 02/2016 |
Description | Collaborative training parntership with AstraZeneca and University of Glasgow |
Amount | £1,000,000 (GBP) |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 11/2016 |
End | 10/2019 |
Description | Syngenta Industry Case studentship |
Amount | £80,000 (GBP) |
Organisation | Syngenta International AG |
Sector | Private |
Country | Switzerland |
Start |
Description | AstraZenenca - data, staff training, papers |
Organisation | AstraZeneca |
Country | United Kingdom |
Sector | Private |
PI Contribution | Provision of data on chemical effects analysis in fish and other animals Training of AZ staff in molecular methods Development of specific test methods Co-authored research papers Shared research programmes |
Collaborator Contribution | Finance Access to research facilities Support for grant applications Training of Exeter staff in GLP and industry environment (secondments) Shared research programmes |
Impact | New test methods for chemicals Strategic partnership Co-authored research papers CASE partnership on PhD projects |
Description | BASF |
Organisation | BASF |
Country | Germany |
Sector | Private |
PI Contribution | Provision of data on nanopoymner effects analysis in fish and other animals Development of specific test methods Co-authored research paper Shared research programmes |
Collaborator Contribution | Funding |
Impact | Research paper Research training Provision of specific test materials ( from BASF) for testing |
Start Year | 2012 |
Description | CEFAS |
Organisation | Centre For Environment, Fisheries And Aquaculture Science |
Country | United Kingdom |
Sector | Public |
PI Contribution | Provision of data on chemical effects analysis in fish and other animals Development of specific test methods Co-authored research papers Shared research programmes Stategic partnership |
Collaborator Contribution | Funding CASE partner on PhD studentships Shared research programmes Stategic partnership |
Impact | PhD graduates Co-authord research papers Co-authored government reports |
Start Year | 2011 |
Description | Syngenta (internships and studentship) |
Organisation | Syngenta International AG |
Country | Switzerland |
Sector | Private |
PI Contribution | Support for PhD student Co-authored research papers Shared research programme |
Collaborator Contribution | Support for PhD student Co-authored research papers Expertise in modelling |
Impact | One PhD graduate (a further PhD student registered) Co-authored research papers |
Start Year | 2009 |
Description | 50th Anniversary of the Fisheries Society of the British Isles |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | The Jack Jones Lecture, 50th Anniversary of the Fisheries Society of the British Isles. Feminisation of Nature - An Unnatural History, Exeter, July 2017. C.R.Tyler Assessing the oestrogenic potency and health impacts of wastewater treatment work effluents using ERE-GFP transgenic zebrafish. FSBI Annual Symposium, Exeter, UK 3-7 July 2017 Oral presentation Cooper, R.; Kudoh, T.; Tyler, C.R.; David, A.; Hill, E. |
Year(s) Of Engagement Activity | 2017 |
Description | 9th Interntionaal Symposium of Reproductive Physiology of Fish Are Chemicals Impacting on Fish Populations? |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Participants in your research and patient groups |
Results and Impact | - |
Year(s) Of Engagement Activity | 2011 |
Description | Chalkstream Headwaters |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | Feminisation of Freshwater Nature - Chalk Stream Headwaters Annual Meeting at Sparsholt College, 24th November 2017 |
Year(s) Of Engagement Activity | 2017 |
Description | International Congress on Comparative Endocrinology, Canada |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | Endocrine Disruption in Aquatic Systems. International Congress on Comparative Endocrinology, Canada. June 2017. Keynote speech. |
Year(s) Of Engagement Activity | 2017 |