Tracking and assessing the Risk from Antibiotic resistant genes using Chip technology in surface water Ecosystems (TRACE) Supervisor: Dr Enda Cummins PhD student: Eithne OFlaherty Date: Page 2 Objective of Irish project Overall Objective: The main objective of the project is to develop a risk assessment model for human exposure to selected Antimicrobial Resistant Organisms (ARO) from surface water use at selected sites Specific focus on the probability of human exposure to AROs from irrigation, drinking water and recreational water Page 3 Tasks Task 1: Literature review and development of risk assessment framework model (Completed) Task 2: Hazard Identification- Data on AROs at selected sites will be collected from EU partners (Ongoing) Task 3: Exposure Assessment- Estimation of the probability of human exposure from recreational, drinking and irrigation practices will be carried out (Ongoing) Task 4: Hazard Characterization- Exploring the human health consequence of the exposure to the AROs Task 5: Risk Characterization- Combining the data found from the previous tasks to give a complete risk assessment. Page 4 Milestones and Deliverables Milestone 1: Literature review and development of a risk assessment framework model- (month 13) (Completed) Milestone 2: Collation of data (from literature, project partners and other agencies) to characterise and model levels of selected ARO in surface water at designated sites - (month 25) (Ongoing) Milestone 3: Analysis of data for use in probabilistic risk assessment model - (month 30) Deliverable: A risk assessment model of human exposure to selected antimicrobial resistant organism from surface water use (month 36) Project Timetable Months Task 1: Literature Review Task 2: Hazard Identification Task 3: Exposure model for selected ARO Task 4: Hazard characterisation for selected ARO Task 5: Risk Characterisation Task 6: Management & Dissemination Milestones M1 M2 M3 Deliverables D1 Currently Page 6 Literature review Antibiotic resistance in surface water ecosystems: presence in aquatic environment, prevention strategies and risk assessment (contributes to completion of task 1) - Question: What factors influence antimicrobial resistant organisms (ARO) in surface water systems and what risk assessment strategies can be used to evaluate the risk from ARO in surface water ecosystems. - It combines a systematic review approach (answering/addressing the research question through the use of literature) with an integrative review approach (where we attempt to find common ideas and concepts from the reviewed material). Page 7 Literature review Introduction Presence in the environment: - Wastewater treatment plants and hospital wastewater - Irrigation - Recreational water - Farm animals - Drinking water - Pristine areas Prevention strategies through water treatments: - Primary and secondary wastewater treatments - Disinfection methods Approaches to Risk Assessment: - Environmental models - Catchment models - Statistical based models - Human health risk assessment Conclusion and recommendations References -Draft provided on SmartSimple -Propose to send for International Peer Review Page 8 Main research findings: - Antibiotic resistance is present at varying levels in wastewater treatment plants (WWTPs), hospital wastewater, irrigation, recreational water and drinking water -Water treatment processes examined: - Chlorination - UV - Ozone - Treatments showed a reduction of antimicrobial resistance but also showed in some circumstances may not have an effect or could increase resistance Literature review Page 9 No gold standard risk assessment method for evaluating antibiotic resistance Examined different types of model strategies : - Environmental models (EUSES, QSAR and ECOSAR) - Catchment models (PhATE, GREAT-ER, GWAVA) - Statistical based models and human health probabilistic risk assessments (based on PEC, PNEC, HQ and MECs) Literature review Page 10 Actions from last meeting Irrigation Synthetic wastewater: Where are the papers based?- see Irrigation section in draft Drinking Water: Need to specify that the literature review has been carried out on a global context. There is a need to specify the type of treatments for drinking water (if available in the papers)- see Drinking water section in draft It would be important to mention the n. of studies- see summary table in draft Recreational Water: There is a need to differentiate lake/beach (wherever available in the papers reviewed)- see Recreational water section in draft Page 11 Sampling Strategy: Spain River Ter -Sampling point taken 1000m upstream of WWTP (3 samples) -Sampling point taken 200m downstream of WWTP(3 samples) -Samples taken in June and October 2015 Page 12 Sampling Strategy: Italy River Tiber and Ostia beach -One sampling point downstream of WWTP -One sampling point where River Tiber enters sea -Two sampling points along Ostia beach (Summer 2016) Page 13 Model development Risk assessment models: Model 1: Data collected from the River Ter (Spain) will examine human exposure to AROs through drinking water Model 2: Data collected from Ostia beach (Italy) will examine human exposure to AROs through recreational water Model 3: Data collected from the River Tiber (Italy) and River Ter (Spain) will examine human exposure to AROs through irrigated crops Page 14 Dinking water model Level in tap water ARO level in river Ter Level before drinking water treatment Level after each water treatment Level consumed by human Environmental parameters influencing ARO level in river An empirical formula will be used to model the environmental influencing factors that may cause decay of ARO (Task 2, developing database in excel) Data from Spanish TRACE project partners (Task 2, some data points collected from partners) UV (meta- analysis study, 200 data points collected) ozonation, chlorination and filtration (Task 2, developing database in Excel) Level of AROs in water before DWTP. Level of AROs in water after water distribution system Amount of AROs water consumed by human through drinking tap water (Task 3, exposure assessment) Page 15 ARB tested at River Ter Antibiotic resistant bacteria tested for at River Ter site are (contributing to task 2): Escherichia coli resistant to Ampicillin Cefotaxime Ciprofloxacin Trimethoprim Page 16 Recreational water model ARO level in Ostia beach Level consumed by human in beach water Exposure to ARO through recreational activity Environmental parameters influencing ARO level in beach Data from Italian TRACE project partners and literature (Task 2, some data collected from literature) Survey created and completed by beach goers at Ostia beach, waiting for data to be sent back (Task 3, exposure assessment) Amount of AROs consumed by human through recreational activity. (Task 2 and Task 3) An empirical formula will be used to model the environmental influencing factors that may cause decay of ARO (Task 2, developing database in Excel) Page 17 ARB tested at Ostia beach Antibiotic resistant bacteria tested for at Ostia beach site are (contributing to task 2): Escherichia coli resistant to Ciprofloxacin Nalidixic acid Amoxicillin Azithromycin Sulfamethoxazole Tetracycline Gentamycin Cefotaxime Imipenem Page 18 Level of AROs after food processing ARO level in river Ter and Tiber Level of ARO in harvested crop Level of ARO in water used for irrigation Level of ARO absorbed by crops Level of ARO in crop at consumption Environmental parameters influencing ARO level in river An empirical formula will be used to model the environmental influencing factors that may cause decay of ARO (Task 2, developing database in excel) Data from Spanish and Italian TRACE project partners (Task 2, some data collected from partners) Type of crops (corn, rice, alfalfa, fruit trees) and amount of ARO absorbed by crop through empirical formula (task 2, developing data in excel) Amount of water used in irrigation, ARO level in irrigation water (task 2, developing data in excel) Level of AROs in harvested crop Effect of any food processing that may contribute to decay/growth rate of bacteria in crop Amount of ARO consumed by human through eating crop (Task 3, exposure assessment) Irrigated crops model Page 19 ARB tested at River Tiber and Ter Antibiotic resistant bacteria tested for at both sites are (contributing to task 2): Escherichia coli resistant to Ciprofloxacin Nalidixic Acid Amoxicillin Azithromycin Sulfamethoxazole Trimethoprim Tetracycline Gentamycin Cefotaxime Imipenem Ampicillin Page 20 Decided with TRACE project partners to examine antibiotic resistant Escherichia coli at the selected sites (contributes to task 2) Development of a database using literature data on the effect of environmental factors (solar radiation, pH, temperature and salinity) on resistant Escherichia coli (task 2 currently working on) Characterisation of each site using literature data and data from project partners Hazard Identification Page 21 Exposure Assessment The TRACE project partners from Italy asked the survey to beach goers at Ostia beach The survey will help calculate the water consumed through different recreational activities (contribute to task 3). Surveys being collated by Italian project partners - data can be analysed and used in risk assessment model Page 22 Exposure Assessment Actions from last meeting: Discussion about the difficulty in establishing the human health consequence as a result of the human exposure to AROs: It was recommended to focus on what the human exposure level was, as it is difficulty to establish the health consequence If we do come across appropriate method of establishing the human health consequence through scientific literature we will review Page 23 Meta-analysis study Went on a 2 week ERASMUS + exchange program to Nantes, France (November 2015) to learn how to do a meta-analysis study A meta-analysis is a statistical method of combining results from comparable studies, this can strengthen a study by giving a weighted average Decided to investigate the effect of UV treatment on sensitive and antibiotic resistant Escherichia coli (data collection contributes to task 2 and drinking water model) Page 24 Meta-analysis study Learned the basics of WinBugs software Since returning, collection of over 200 data points on the effect of UV treatment on sensitive and antibiotic resistant Escherichia coli Preliminary findings (not all 200 data points included yet) show that different Escherichia coli strains have different log reductions patterns from UV treatment (see next slide) Slope Page 26 With the completion of the meta-analysis model: The effect of UV treatment for a log reduction of both sensitive E. coli and antibiotic resistant E. coli established Investigate if different strains of Escherichia coli have different log reduction patterns from UV treatment Can use this data in drinking water model to see how resistant E. coli is effected by UV water treatment (task 2) Meta analysis can be used to study other process effects (e.g. filtration) Meta-analysis study Page 27 Conferences Will be presenting at the Biosystems and Food Engineering Research Day in UCD (March 2016) Propose to present a poster at ENVIRON 2016 in University of Limerick (March 2016) Propose to present a poster at the 13 th IWA Leading Edge Conference on Water and Wastewater technologies in Jerez de la Frontera, Spain (June 2016) Details uploaded to SmartSimple Page 28 Training 2 week ERASMUS + exchange program to Nantes, France to learn how to do a meta-analysis study (November 2015) Had first doctoral studies panel meeting where I presented about the TRACE project (January 2016) Page 29 Tasks and milestones for next 6 months Task 2: Continued progress on data collection of AROs at selected sites from EU project partners and development of hazard identification database (month 7-15) Task 3: Continued progress on the estimation of the probability of human exposure from recreational and irrigation practices and to identify the level and survival behaviour of the ARO (month 10-30) Milestone 2: Collation of data (from literature, project partners and other agencies)to characterise and model levels of selected ARO in surface water at designated sites(month 20) Page 30 Skills learned A two week ERASMUS course (23rd of March- 3rd of April 2015) was completed. Learned how to use software that will be used in work package 6 to build the risk assessment model (Result A-) A Quantitative Risk Assessment module was completed in UCD (January- May 2015), valuable information about quantitative risk assessment techniques, methods and software were learned (Result A+) A four page paper on the Identification of antimicrobial resistant organisms in surface water ecosystems and risk assessment strategies for their control was published and presented for the Biosystems Engineering review book 20 (July 2015)ew_20 Thank you!