Form 101 - Application for a Grant Send to NSERC with your attachments ...ecobim.ca/files/2017/03/IPOC-CRSNG_2012.pdf · Send to NSERC with your attachments, if applicable Applicant:

Send to NSERC with your attachments, if applicable Form 101 - Application for a Grant

Applicant:

Program:

Application Title:

175797686

Michel Fournier

Strategic Project Grants

INRS

Interaction between pollution and climate changes: development of improved monitoring strategy

Reference Number:

For NSERC office use only

14327NSERC PIN:

Michel FournierForm 101 - Application for a Grant

Electronic Attachments:Budget Justification - Budget justificationContributions from Supporting Organizations - Attachment - IPEVQuotations - Quotations 2012Proposal - Proposal IPOCReferences - ReferencesRelationship To Other Research Support - RelationshipOther Documents - CV

Michel FournierF100/Personal Data Form

Electronic Attachments:Contributions - PDF File

Pauline BrousseauF100/Renseignements personnels

Electronic Attachments:Contributions - Contributions 03 2012

Daniel CyrF100/Personal Data Form

Electronic Attachments:Contributions - Contributions

François GagnéF100/Renseignements personnels

Electronic Attachments:Contributions - Liste de publications

FREDERICK KIBENGEF100/Personal Data Form

Electronic Attachments:Contributions - CONTRIBUTIONS

Page 1 of 2

Send to NSERC with your attachments, if applicable

Applicant:

Program:

Application Title:

175797686

Michel Fournier


INRS


Reference Number:

For NSERC office use only

14327NSERC PIN:

Ahmed SiahF100/Personal Data Form


Philip SpearF100/Renseignements personnels

Electronic Attachments:Contributions - contributions scientifiques P. Spear

Yves St-PierreF100/Renseignements personnels


Cathy VaillancourtF100/Renseignements personnels

Electronic Attachments:Contributions - Contributions-Mars-2012

Michel FournierForm 183A - Information Required from Organizations Participating in RPP (submitted by applicant)

Electronic Attachments:Letter(s) of Support - IMLLetter(s) of Support - CEAEQLetter(s) of Support - AquariumLetter(s) of Support - BiodomeLetter(s) of Support - MakivikLetter(s) of Support - ECLetter(s) of Support - CIRELetter(s) of Support - Lab Bell

Page 2 of 2

173,560290,848

CERTIFICATION/REQUIREMENTS

SIGNATURES (Refer to instructions "What do signatures mean?")

TOTAL AMOUNT REQUESTED FROM NSERC

For Strategic Projects, indicate the Target Area and the ResearchTopic; for Strategic Networks indicate the Target Area.

Given name

MichelInitial(s) of all given names

Type of grant applied for

Family name of applicant

Time (in hours per month) to be devoted to the proposed research / activity

Year 1 Year 2 Year 3 Year 4 Year 5

Head of departmentApplicantApplicant's department, institution, tel. and fax nos., and e-mail

Form 101 (2011 W)

20

MPersonal identification no. (PIN)

President of institution(or representative)

Dean of faculty


NO If YES to either question a) or b) – Appendices A and B must be completedX

Does any phase of the research described in this proposal a) take place outside an office or laboratory, or b) involve an undertaking as describedin Part 1 of Appendix B?

Primary

1501

Secondary

5504

Primary Secondary

402

Research subject code(s)

406

Area of application code(s)

climatic and habitat changes, freshwater and marine models, pollution, bivalves, immnunotoxicology, genomic, ecotoxicologic, physiology, cell biology, chemistry

If this proposal involves any of the following, check the box(es) and submit the protocol to the university or college's certification committee.

BiohazardsHumans

It is agreed that the general conditions governing grants as outlined in the NSERCto this application and are hereby accepted by the applicant and the applicant's employing institution.

Program Guide for Professors apply to any grant made pursuant

Head Office

0 0

Environmental Science and Technologies / Enhancing Aquatic Ecosystem Services

Research involving : Human pluripotent stem cells Animals X

PROTECTED WHEN COMPLETED

The information collected on this form and appendices will be storedin the Personal Information Bank for the appropriate program.

Version française disponible

INRSTel.: (450) 687-5010 ext. 8824FAX: (450) [email protected]

Language of application

English FrenchX

Institution that will administer the grantINRS

Provide a maximum of 10 key words that describe this proposal. Use commas to separate them.

Interaction between pollution and climate changes: development of improved monitoring strategyTitle of proposal

Fournier

2012/04/01Date

System-ID (for NSERC use only)

175797686

Institutional Identifier

14327Valid

FORM 101Application for a Grant

PART I

225,958

CO-APPLICANTS

2

I have read the statement "What do signatures on the application mean?" in the accompanying instructions and agree to it.Research/

activity time(hours/month)

Personal identification no. (PIN) Family name of applicant

PIN, family name and initial(s) Organization Signature

Fournier14327Valid

115598, Cyr, DG INRS

264259, Vaillancourt, C INRS

115621, Gagné, FG Québec à Rimouski

336100, Siah, A Prince Edward Island

323481, Brousseau, P Québec à Rimouski

174521, St-Pierre, STPY INRS

56653, Spear, A Québec à Montréal

56657, Kibenge, S.B. Prince Edward Island

Use additional page(s) if necessary.Form 101 (2011 W), page 2 of 9 Version française disponible


CO-APPLICANTS' ORGANIZATIONS AND/OR SUPPORTING ORGANIZATIONS (if organization different from page 1)

Martineu, Pascale, PMAgente de recherche et de planificationQuébec à Montréal

Schultz, Katherine, KSVice-President Research & DevelopmentPrince Edward Island

Demers, Serge, SDDirecteurISMER à l'UQAR

2 - 2

Use additional page(s) if necessary.Form 101 (2011 W), page 2 - 2 of 9 Version française disponible



Fournier14327Valid

Family name and given name of signing officer,title of position, and name of organization Signature

pursuant to this application and are hereby accepted by the organization.

It is agreed that the general conditions governing grants as outlined in the NSERC Program Guide for Professors, as well as the statements "What do signatures on the application mean?" and "Summary of proposal for public release" in the accompanying instructions, apply to any grant made

Kwan, MK Makivik Corporation,204351, Lebeuf, ML Maurice Lamontagne Institute,Bellemare, FB Lab_Bell Inc.,de Lafontaine, YDL Centre Saint-Laurent,Masson, SM Aquarium du Québec,Vaillancourt, RB Biodôme de Montréal,Martel, LM CEAEQ,

COLLABORATORS

2 - 1 Collaborators (RPP except SNG)


Use additional page(s) if necessary. Version française disponible

PROTECTED WHEN COMPLETEDForm 101 (2011 W), page 2 - 1 of 9

FournierBefore completing this section, read the instructions for the definition of collaborators in the Eligibility Criteria section of the Program Guide for Professors.

PIN, family name and initial(s)Research/

activity time(hours/month)

Organization / Department

14327Valid

3

SUMMARY OF PROPOSAL FOR PUBLIC RELEASE (Use plain language.)

Family name of applicantPersonal identification no. (PIN)

Business telephone no. (optional):

E-mail address (optional):

1 (450) 6875010 Ext. [email protected]

This plain language summary will be available to the public if your proposal is funded. Although it is not mandatory, you may choose toinclude your business telephone number and/or your e-mail address to facilitate contact with the public and the media about your research.


La pression anthropique provoque des effets délétères profonds sur les écosystèmes aquatiques côtiers, dont la production est d'une importance capitale pour l'homme. Les changements climatiques en cours sont censés générer dans un futur proche, des conditions environnementales stressantes inconnues, qui vont probablement affecter profondément la biota d'écosystèmes ciblés comme les estuaires. Ces habitats sont naturellement très variables, à la fois spatialement et temporellement. Ces conditions environnementales rendent ces écosystèmes très exposés aux effets synergiques de la pollution et le stress lié aux changements de température. Les invertébrés benthiques, en produisant une énorme quantité de biomasse, contrôle la production des écosystèmeset devraient donc être considérés comme des populations à risque. IPOC vise à fournir à la communauté scientifique et aux gestionnaires de l'environnement une « trousse à outils » améliorée, basée sur des connaissances scientifiques novatrices, afin d'évaluer les effets biologiques de l'action combinée des principauxfacteurs de stress anthropiques des milieux aquatiques, la pollution et les changements climatiques. L'enjeu sera de mieux évaluer la sensibilité d'espèces sentinelles dans un continuum eau douce - eau salée de même queleur vulnérabilité face aux stress ciblés. L'approche combinant l'observation et l'expérimentation permettra de:- Caractériser l'évolution temporelle de l'état de santé des populations d'invertébrés en mesurant une sélection de réponses biologiques au niveau de l'individu par le biais d'une batterie innovante de marqueurs moléculaires,cellulaires et physiologiques;- Prévoir la capacité des populations à risque à se maintenir dans un environnement modifié par le changementglobal;- Proposer une base opérationnelle pour des programmes scientifiques afin de permettre des suivis à long terme conformément aux recommandations d'OSPAR.

Anthropogenic pressure (habitat destruction, chemical contamination, eutrophication) has deep deleterious effects on coastal aquatic ecosystems whose production is of major importance for humans. Ongoing climate changes are expected to generate in a next future unexperienced, stressful environmental conditions which will probably deeply affect biota in targeted habitat as estuaries. These habitats are naturally highly variable, both spatially and temporally. Such environmental conditions make them highly exposed to synergistic effects of pollution and temperature stress. At the basis of the trophic chain, benthic invertebrates, by producing a huge amount of biomass, control ecosystem production and should now be considered as populations at risk. IPOC aims to provide the scientific community and environmental managers with an improved "tool-kit", based on original scientific knowledge, for assessing biological effects of the combined action of the main anthropogenic stressors of aquatic environments, pollution and climatic changes. IPOC scientific rationale relies on a better understanding of the sensitivity of sentinel species in the continuum freshwater to marine environment and thus their vulnerability to targeted stressors. The approach combining observation and experimentation will allow to: - characterize the temporal evolution of the health status of invertebrate populations by measuring a selection of biological responses at the individual level through an innovative battery of molecular, cellular and physiological markers; - foresee the capacity of populations at risk to maintain themselves in an environment altered by global change;

- propose an operational basis for scientific programs to allow further long-term monitoring according to OSPAR recommendations;

Other Language Version of Summary (optional).

Fournier14327Valid

PROTECTED WHEN COMPLETEDForm 101 (2011 W), page 3 of 9

ACTIVITY SCHEDULE


Description of activitiesAnticipated

starting dateAnticipated

completion date

(Refer to instructions to see if this section applies to your application. Use additional page(s) if necessary.)

Milestone

4 (RPP)

Fournier14327Valid

2012-11-01 2015-05-31For each proposed sites enough bivalves to obtain the necessary mass of tissues to perform the suite of chemicalanalyses will be collected and adequately preserved until analysis.

Task 5: Assessing the body burden of pollutants

2012-11-01 2014-12-31For this work package, we will benefit from the experiments described in WP1. At each selected time-point, hemocytes will be collected and exposed in a dose-response manner to selected model toxicants.

Task 3: Assessment of sensitivity to in vitro model exposure

2012-11-01 2014-12-31This first part of this research program has been designed to monitor the impacts of changes of various habitat conditions (water temperature, salinity, pH) on physiological systems.

Task 1: Assessment of biomarkers to habitat changes

2013-03-01 2013-11-30This work package will benefit from the experiments described in WP2. At both selected time-points, hemocytes will be collected from 10 individuals from each site and exposed in a dose-response manner to model toxicants.

Task 4: Assessment of sensitivity of endogenous organism

2013-03-01 2013-11-30This task is designed to assess differences in physiological competence of bivalves according to sites location and history.

Task 2: Sites selection based on physiological competence

2014-05-01 2015-05-31The final experimental section of the project will involve an annual follow-up of stocks of wild bivalves collected from both marine and freshwater selected sites.

Task 6: Field validation

2014-11-01 2015-10-31The two main objectives of this task are: -through inter-laboratory approach, i) identification and classification of the most relevant markers (individual and population levels) for the characterization of temporal evolution of the health status of popu

Task 7: Global analysis of data

PROTECTED WHEN COMPLETEDForm 101 (2011 W), Version française disponiblepage 4 of 9

PROPOSED EXPENDITURES FOR DIRECT COSTS OF RESEARCH (include cash expenditures only)

Salaries and benefits

100,980 0 0187,560187,560Students

35,000 0 0070,000Postdoctoral fellows

0 0 000Technical/professional assistants

0 0 000Equipment or facility

0 0 0056,890Purchase or rental

0 0 000Operation and maintenance costs

0 0 000User fees

74,000 0 079,20094,200Materials and supplies

Travel

8,000 0 06,0002,000Conferences

10,000 0 0207,822207,822Field work

4,000 0 04,0004,000Collaboration/ consultation

Dissemination costs

9,000 0 03,0000Publication costs

0 0 000Other activities

Other (specify)

0 0 000

0 0 000


1)

3)

4)

a)

b)

c)

TOTAL PROPOSED EXPENDITURES00240,980487,582622,472

Water quality, pollution

5Personal identification no. (PIN)

Total cash contribution fromindustry (if applicable)

Total cash contribution fromuniversity (if applicable)

Total cash contribution fromother sources (if applicable)

TOTAL AMOUNT REQUESTED FROM NSERC (transfer to page 1)

Form 101 (2011 W), page 5 of 9 Version française disponiblePROTECTED WHEN COMPLETED

0 0 0 0 0

110,710 110,710 67,420 0 0

220,914 150,914 0 0 0

a)

b)

c)

d)

2)

a)

b)

c)

5)

a)

b)

6)

a)

b)


Before completing this section, read the instructions and consult the Use of Grant Funds section in the NSERC Program Guide for Professors for information about the eligibility of expenditures for the direct costs of research and the regulations governing the use of grant funds. On separate page(s), supply a detailed explanation, and justification, for your proposed expenditures. Also explain the relationship or difference between this application and all other research support (held or applied for), and describe any contributions from other sources (if applicable).

290,848 225,958 173,560 0 0

Fournier14327Valid

PROPOSED EXPENDITURES FOR DIRECT COSTS OF RESEARCH (include cash expenditures only.)

70,000 0 35,000 0 0

0 0 0 0 0

0 0 0 0 0

56,890 0 0 0 0

0 0 0 0 0

0 0 0 0 0

94,200 79,200 74,000 0 0

2,000 6,000 8,000 0 0

207,822 207,822 10,000 0 0

0 3,000 9,000 0 0

0 0 0 0 0

4,000 4,000 4,000 0 0

0 0 0 0 0

0 0 0 0 0

187,560 187,560 100,980 0 0

Form 101 (2011 W), page 5 of 9 Version française disponible


a)

b)

c)

d)

a)

Students

Postdoctoral fellows

Technical/professional assistants

1) Salaries and benefits

b)

c)

Purchase or rental

Operation and maintenance costs

User fees

2) Equipment or facility

3) Materials and supplies

4) Travel

a) Conferences

b) Field work

c) Collaboration/consultation

5) Dissemination costs

a) Publication costs

b) Other activities

6) Other (specify)

b)

a)

TOTAL PROPOSED EXPENDITURESFOR DIRECT COSTS OF RESEARCH

Total cash contribution fromindustry (if applicable)

Total cash contribution fromuniversity (if applicable)

Total cash contribution fromother sources (if applicable)

TOTAL AMOUNT REQUESTED FROM NSERC (transfer to page 1)

220,914 150,914 0 0 0

110,710 110,710 67,420 0 0

0 0 0 0 0

622,472 487,582 240,980 0 0

5



Before completing this section, read the instructions and consult the Use of Grant Funds section in the NSERC Program Guide for Professors forinformation about the eligibility of expenditures for the direct costs of research and the regulations governing the use of grant funds. On separate page(s), supply a detailed explanation, and justification, for your proposed expenditures. Also explain the relationship or difference between this application and all other research support (held or applied for), and describe any contributions from other sources (if applicable).

290,848 225,958 173,560 0 0

Sum Total

Fournier14327Valid

6

0 0 0 0 0

0 0 0 0 0

150,000 0 0 0 0

0 0 0 0

0 0 0 0 0

0 0 0 0 0

75,000 75,000 75,000 0 0

25,000 25,000 25,000 0 0

0 0 0 0 0

150,000 0 0 0 0

110,710 110,710 67,420 0 0

INRSCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS


Cash contributions to direct costs of research (Transfer amounts to page five (5);except those for the Ship Time program.)

Personal identification no. (PIN)

Form 101 (2011 W), page 6 of 9 PROTECTED WHEN COMPLETED Version française disponible


section in the NSERC Program Guide for Professors concerning the eligibility of expenditures for the direct costs of research and the regulations

Name of supporting organization

Use of Grant Funds

4)

5)

6)

1)

2)

3)

In-kind contributions to direct costs ofresearch

Salaries for scientific and technicalstaff

Donation of equipment, software

Donation of material

Field work logistics

Provision of services

governing the use of grant funds regarding the eligibility of in-kind contributions.

, and Guidelines for Evaluating Cost-Sharing Ratios and In-Kind Contributions in University-Industry

Before completing this section, read the instructions for contributions from supporting organizations and consult the

Collaborations

Total of in-kind contributions to direct costs of research

1)

2)

3)

In-kind contributions to indirect costs of research (not leveraged)

0 0 0 0 0

250,000 100,000 100,000 0 0

Use of organization's facilities

Salaries of managerial andadministrative staff

Total of all in-kind contributions

Contribution to postsecondary institution overhead

Fournier14327Valid

6 - 1

30,000 30,000 30,000 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

30,000 30,000 30,000 0 0

0 0 0 0 0

Centre Saint-Laurent, Environnement CanadaCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS








Use of Grant Funds

4)

5)

6)

1)

2)

3)










Collaborations


1)

2)

3)


0 0 0 0 0

30,000 30,000 30,000 0 0





Fournier14327Valid

6 - 2

12,500 12,500 12,500 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

8,500 14,500 9,500 0 0

0 0 0 0 0

0 0

0 0 0 0 0

0 0 0 0 0

21,000 27,000 22,000 0 0

0 0 0 0 0

Institut Maurice LamontagneCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS








Use of Grant Funds

4)

5)

6)

1)

2)

3)










Collaborations


1)

2)

3)


0 0 0 0 0

21,000 27,000 22,000 0 0





Fournier14327Valid

6 - 3

100,000 100,000 100,000 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

50,000 50,000 50,000 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

150,000 150,000 150,000 0 0

70,000 0 0 0 0

Centre interinstitutionnel de recherche en écotoxicologieCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS








Use of Grant Funds

4)

5)

6)

1)

2)

3)










Collaborations


1)

2)

3)


0 0 0 0 0

150,000 150,000 150,000 0 0





Fournier14327Valid

6 - 4

10,000 10,000 10,000 0 0

0 0 0 0 0

8,900 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

18,900 10,000 10,000 0 0

0 0 0 0 0

Lab-Bell IncCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS








Use of Grant Funds

4)

5)

6)

1)

2)

3)










Collaborations


1)

2)

3)


0 0 0 0 0

18,900 10,000 10,000 0 0





Fournier14327Valid

6 - 5

30,000 30,000 30,000 0 0

0 0 0 0 0

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0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

30,000 30,000 30,000 0 0

0 0 0 0 0

Biodôme de MontréalCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS








Use of Grant Funds

4)

5)

6)

1)

2)

3)










Collaborations


1)

2)

3)


0 0 0 0 0

30,000 30,000 30,000 0 0





Fournier14327Valid

6 - 6

60,000 30,000 30,000 0 0

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60,000 30,000 30,000 0 0

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Aquarium du QuébecCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS








Use of Grant Funds

4)

5)

6)

1)

2)

3)










Collaborations


1)

2)

3)


0 0 0 0 0

60,000 30,000 30,000 0 0





Fournier14327Valid

6 - 7

16,500 16,500 16,500 0 0

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0 0 0 0 0

10,000 10,000 10,000 0 0

30,000 30,000 30,000 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

56,500 56,500 56,500 0 0

0 0 0 0 0

Makivik CorporationCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS








Use of Grant Funds

4)

5)

6)

1)

2)

3)










Collaborations


1)

2)

3)


0 0 0 0 0

56,500 56,500 56,500 0 0





Fournier14327Valid

6 - 8

16,800 16,800 16,800 0 0

0 0 0 0 0

0 0 0 0 0

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15,000 25,000 13,000 0 0

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0 0 0 0 0

0 0 0 0 0

31,800 41,800 29,800 0 0

0 0 0 0 0

Centre d'expertise en analyse environnementale du QuébecCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS








Use of Grant Funds

4)

5)

6)

1)

2)

3)










Collaborations


1)

2)

3)


0 0 0 0 0

31,800 41,800 29,800 0 0





Fournier14327Valid

6 - 9

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

136,869 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

0 0 0 0 0

136,869 0 0 0 0

150,914 150,914 0 0 0

Institut Polaire FrançaisCONTRIBUTIONS FROM SUPPORTING ORGANIZATIONS








Use of Grant Funds

4)

5)

6)

1)

2)

3)










Collaborations


1)

2)

3)


0 0 0 0 0

136,869 0 0 0 0





Fournier14327Valid

BUDGET JUSTIFICATION

Salaries and benefits. In this proposal, a certain number of technicians and research

professionals are involved for the monitoring of biomarkers, sampling of organisms and

maintenance of animals during exposure experiments. No budget is requested from

NSERC since all salaries and benefits are covered by the different partners involved in

the program (see supporting letters). These contributions represent a financial support to

the proposal estimated at 275,800$ annually. We are only requesting from NSERC

financial support for students bursaries.

Bursaries. According to INRS and other universities, every student registered in

graduate programs must be financially supported. Minimum contribution for a M.Sc.

student is 14,430$ and 16,830$ for a PhD student. Most of the universities contribute for

up to 50% of the bursaries for a period of 2 and 3 years for a M.Sc. and a PhD student

respectively. Therefore, in this proposal we are requesting from NSERC the other half of

bursaries. All M.Sc. students (6) will be involved the first 2 years of the program,

therefore: 6 x 7,215$ = 43,290$/year. All PhD students (6) will be involved the three

years of the program therefore: 6 x 8,415$ = 50,490$/year. The two post-doctoral

fellows involved the first year of the program are covered by a contribution from CIRÉ (2

x 35,000$), we are requesting a bursary for the post-doctoral fellow involved in the third

year (35,000 $). Finally two French PhD students will be involved in the program, their

bursaries will be supported by French Universities and INRS (2 x 8,415$ =

16,830$/year).

Equipment. One piece of equipment, a portable flow cytometer, is requested for the

program. In the last three decades, our group has been particularly active in developing

the use of flow cytometry to monitor the impact of pollutants at cellular and molecular

levels. We were able to adapt this tool for the determination of adverse effects of

chemicals and drugs on cellular parameters (cell cycle, apoptosis, necrosis), immune

endpoints (phagocytosis, NK activity…) and metabolic markers (metallothionein,

esterase…). These developments have generated a considerable number of scientific

publications (see M. Fournier’s CV) and books (Handbook of immunological methods,

Brousseau et al., 1999; Investigative Immunotoxicology, Tryphonas et al. 2005).

Moreover, in the last decade, using portable flow cytometer, we were able to adapt our

methodologies to field conditions, allowing monitoring of endpoints on the sites. To

proceed with all the requirements of equipment for the present project with sampling sites

spread in several areas (Nunavut, Great Lakes, Eastern Quebec, Kerguelen Islands…) we

will need a second portable flow cytometer. We have performed an evaluation of several

systems from different companies (Guava, Cederlane, Partec) and the most robust and

performing system is C6 Flow Cytometer System from Becton Dickinson. This system

totally digitalized obviates the need for adjustable gain settings on the detectors. Data is

collected and always remains in the data file eliminating the need to set gain or voltage,

which reduces the consumption of valuable sample and sheath during set-up. The cost of

the system is 53,200$ and with applicable taxes the total cost is 56,890 $.

All other pieces of equipment needed are already in place in co-applicants and

collaborators laboratories. We must signaled a contribution from INRS to support a field

station in Parc de la Rivière Mitis for 150,000$ (benches and small equipment) and a

contribution from IPEV for equipment in Kerguelen Island (136,869$)

Materials and supplies. The following table presents the need for materials and supplies

for the various sectors of the research program. Most of the items represent consumables,

standards, probes etc. An amount of 15 000$ has been calculated to purchase aquariums

in year 1. Items Year 1 Year 2 Year 3

Algae 52L/year x

$100/L

5200

5200

0

Aquariums 15000 0 0

Chemicals 25000 25000 25000

Probes 5000 5000 5000

Licences for software 2000 2000 2000

Molecular biology 13000 13000 13000

Immunology 10000 10000 10000

Neuro/endo/repro 9000 9000 9000

Metabolism 10000 10000 10000

Total 94200 79200 74000

Travels. Three types of travel expenses have been budgeted in this proposal. The first

one is to cover the participation of students to scientific meetings in order for them to

present their results. The list of meetings includes SETAC and the annual meeting of the

Canadian Society of Toxicology (2 000$, year 1; 6 000$ year 2; 8 000$, year 3).

The second part of this budget is for the field work in Canada and Kerguelen Islands. For

Canadian sites, a budget of 18,000$ (years 1 and 2) and 10,000$ (year 3) will cover

travels and subsistence fees for students on sampling sites in the Great Lakes, Nunavut as

well as Eastern Quebec and Prince Edward Island. The fees related to the field work in

Kerguelen Island will be covered by two sources. The French Polar Institute (see IPEV’s

Director’s letter), puts a cash contribution of 150,914$ for years 1 and 2. We have to

support through NSERC an amount of 38,908$ for various fees not supported by IPEV

(see letter from IPEV). Total cost for field work is therefore 207,822$ (years 1 and 2) and

10,000$ (year 3). A contribution of 56,908$ (years 1 and 2) and 10,000$ (year 3) is

requested from NSERC.

Finally a provision of 4,000$ is budgeted to cover the participation of students to an

annual meeting of the research team. This meeting will also involve participation of the

French collaborators. It will be hold in conjunction with ÉcoBIM annual meeting of

CIRÉ.

Publications. A budget of 3,000$ and 9,000$ has been planned for the second and third

years of the program to cover publication costs in journals with referees. This budget will

be reserved for publications in which students are first authors.

BD Biosciences 2100 Derry Road West, Suite 100 Mississauga, Ontario, L5N 0B3 tel: 1.888.259.0187 fax: 1.866.542.6659

QUOTATION #2012-033DG Expiry Date: June 15, 2012

Becton Dickinson Canada Inc. Page 2 of 5

To: INRS-Institut Armand-Frappier, Laboratoire Michel Fournier Date: March 2, 2012 531 Boul des Prairies,Laval. FOB: Origin Laval, H7V 1B7 Québec

Attn: Marlène Fortier, RT, M. Sc. Terms: Net 30 Days

Ph: 450-687-5010 Ext: 8852 Currency: $CDN E: [email protected] Research

Mat. No. Description Unit List

Price Disc. 1 (Inst.)

Qty Total Net Price

653118 C6 Flow Cytometer® -Instrument, workstation, monitor -CFlow® Plus Software: offers enhanced capabilities including sample volume tracking, event concentration, fine core size and flow rate controls and time as a parameter

$66,500 ($13,300.00) 1 $53,200.00

Total System List Price (before taxes) $66,500.00 Total Discount 1 (Institutional) ($13,300.00)

Total System Net Price (before taxes) $53,200.00

Options

Mat. No. Description Unit List Price

Disc. 2 (Inst.)

Qty Total Net Price

Optional System Modules

645447 FCAP Array™ v1.0.2(Mac OS) $1,895 ($189.50) 1 $1,705.50 641488 FCAP Array™ v1.0.2(PC Windows) $1,895 ($189.50) 1 $1,705.50

653122 CFlow Plus Analysis Software for PC or Mac (additional license) CFlow Plug Analysis software for use with the CFlow Plus and CFlow (PC or Mac)

$1,000 ($200.00) 1 $800.00

653126 Module for Selectable Lasers (Requires CFlow Plus of CFlow Sampler Software) Facilitates standard three detectors off of blue laser, & one off of red laser. Allows for four blue detectors, or two blue and two red detectors. Includes Activation Key & Optical filters: 780/60, 610/20, 630/30

$8,153 ($815.30) 1 $7,337.70

653127 Module for Cflow Plus User Tracking (Requires CFlow Plus of CFlow Sampler Software) For the creation of unique user names and passwords to record the duration each operator uses the C6. A log file is created, allowing the preparation of billing reports for multi-user or core laboratories

$858 ($85.80) 1 $772.20

CM-70 Enhanced Analysis Module Optional upgrade for CFlow Plus and CFlow Sampler software. Enables event coloring, live gating, batch analysis and exporting of high resolution graphics for publication purposes

$2,500 ($250.00) 1 $2,250.00

653124 CSampler® -96 well plate & 24 tube rack automated sampler for C6 Flow Cytometer -CFlow Plus software upgrade (CF-01) is required for users of CFlow

$31,800 ($6,360.00) 1 $25,440.00

page 2 of 4

The New Dimension in Flow Cytometry

Item # CyFlow® Cube 8 List Price Discount Price

CY-S-3068-488-20_5P

CyFlow® Cube 8 with a 20mW 488nm blue laser and 5 PMTs for forward and side light scatter and 3-color fluorescence detection. $55,900 $41,925

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CyFlow® Cube 8 with a 20mW 488nm blue laser and a 25mW 638nm red laser and 6 PMTs for forward and side light scatter and 4-color fluorescence detection

$73,900 $55,425

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CyFlow® Cube 8 with a 20mW 488nm blue laser and a 25mW 638nm red laser and 8 PMTs for forward and side light scatter and 6-color fluorescence detection

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CY-S-3068-488-20_638-25_UV-

LED_8P

CyFlow® Cube 8 with a 20mW 488nm blue laser, a 25mW 638nm red laser, and a UV-LED and 8 PMTs for forward and side light scatter and 6-color fluorescence detection

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CY-S-3068-488-20_638-25_407-

100_8P

CyFlow® Cube 8 with a 20mW 488nm blue laser, a 25mW 638nm red laser, and a 100mW 407nm violet laser and 8 PMTs for forward and side light scatter and 6-color fluorescence detection

$121,900 $91,425

CY-S-3068-488-20_638-25_375-

16_8P

CyFlow® Cube 8 with a 20mW 488nm blue laser, a 25mW 638nm red laser, and a 16mW 375nm UV laser and 8 PMTs for forward and side light scatter and 6-color fluorescence detection

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CY-S-3068-488-20_638-25_561-

100_8P

CyFlow® Cube 8 with a 20mW 488nm blue laser, a 25mW 638nm red laser, and a 100mW 561nm yellow laser and 8 PMTs for forward and side light scatter and 6-color fluorescence detection

$131,900 $98,925

CyFlow® Cube 8 The CyFlow® Cube 8 measures forward and side light scatter and up to 6 colors of fluorescence. The CyFlow® Cube can be configured with a UV-LED and up to 3 solid-state lasers selected from blue (488nm), green (532nm), violet (407nm), ultraviolet (355, 365 or 375nm), yellow (561nm), orange (594nm), or red (638nm). This compact, desk-top instrument has a 20” by 19” footprint and is equipped with a built-in PC computer with a built-in 19” LCD monitor.

Item # Options for CyFlow® Cube 8 List Price Discount Price

CY-S-3080-8 CyFlow® Robby Autoloading Station for CyFlow® Cube 8 for automated sample input from 96-well microtiter plates and sample tubes $24,000 $18,000

CY-S-P additional PMT for CyFlow® Cube 8 $8,000 $6,000

CY-S-RED40 Upgrade 25mW 638nm laser with a 40mW, 640 nm red diode laser $6,000 $4,500

CY-S-BLUE50 Upgrade 20mW 488nm laser with a 50mW, 488 nm blue solid state laser $8,000 $6,000

CY-S-UVLED 365nm ultraviolet light-emitting diode $10,000 $7,500

page 3 of 4

The New Dimension in Flow Cytometry

Item # CyFlow® Cube 8 Sorter List Price Discount Price

CY-S-3069-488-20_5P

CyFlow® Sorter with a 20mW 488nm blue laser and 5 PMTs for light scatter and 3-color fluorescence detection and includes the piezo-based CyFlow® Sorter (PPCS) for sorting cells and other particles.

$124,160 $93,120

CY-S-3069-488-20_638-25_5P

CyFlow® Sorter with a 20mW 488nm blue laser and a 25mW 638nm red laser and 5 PMTs for light scatter and 3-color fluorescence detection and includes the piezo-based CyFlow® Sorter (PPCS) for sorting cells and other particles.

$134,160 $100,620

The CyFlow ® Cube 8 can be equipped with Partec’s patented, piezo-based sorting module (PPCS). The PPCS is a closed fluidic system, making it ideal for sorting biohazardous material, and is gentle enough to sort even fragile cells. The CyFlow® Cube 8 Sorter configuration can accommodate up to 2 lasers and includes 5 PMTs for FSC, SSC and 3-color fluorescence detection.

CyFlow® Cube 8 Sorter

Item # Installation and Training List Price Discount Price

PNA-09-618 Installation and on-site training for CyFlow® Cube 6, 8, and Sorter $2,600 $1,950

Item # Options for CyFlow® Cube 8 Sorter List Price Discount Price

CY-S-3080-8 CyFlow® Robby Autoloading Station for CyFlow® Cube 8 Sorter for automated sample input from 96-well microtiter plates and sample tubes $24,000 $18,000

CY-S-RED40 Replace 25mW 638nm laser with a 40mW, 640 nm red diode laser $6,000 $4,500

CY-S-BLUE50 Replace 20mW 488nm laser with a 50mW, 488 nm blue solid state laser $8,000 $6,000

CY-S-UVLED 365nm ultraviolet light-emitting diode $10,000 $7,500

CY-S-VIOLET100 Replace 638nm laser with a 100mW, 405nm violet laser $22,000 $16,500

CY-S-UV375 Replace 638nm laser with a 16mW, 375nm UV laser $24,000 $18,000

CY-S-YELLOW100 Replace 638nm laser with a 100mW, 561 nm yellow solid state laser $32,000 $24,000

Phone: 856-467-0018 ● Toll-free: 888-808-0067 ● Fax: 856-467-0188 ● [email protected]

The CyFlow® SL is a portable volumetric flow cytometer that measures forward and side light scatter and up to 3 colors of fluorescence. This versatile instrument can be used for many different applications including immunofluorescence, DNA content for ploidy analysis and cell cycle, fluorescent protein detection and the determination of absolute cell counts and cell concentrations. Accurate counts are achieved by Partec’s TVAC volumetric counting system without the use of expensive reference counting beads or parallel analysis on a hematology counter. The CyFlow® SL is a single laser instrument that can be configured with a solid-state blue (488nm), green (532nm), violet (405nm) or red (635nm) laser. This compact, portable instrument has a 17” by 15” footprint and can be equipped with either a laptop or a desktop computer. It can be powered in the laboratory by 110 volts or 220 volts and in the field by 12 volts DC (i.e.: from a car battery).

Catalog # CyFlow SL Blue List Price

Discount Price

CY-S-1035 CyFlow SL Blue with a 20mW, 488nm, blue laser and 5 PMTs for detection of forward and side light scatter and 3 colors of fluorescence $79,640 $59,730

CY-S-1034 CyFlow SL Blue with a 20mW, 488nm, blue laser and 4 PMTs for detection of forward and side light scatter and 2 colors of fluorescence $71,640 $53,730

CY-S-1033 CyFlow SL Blue with a 20mW, 488nm, blue laser and 3 PMTs for detection of forward and side light scatter and 1 color of fluorescence $63,640 $47,730

CY-S-1032 CyFlow SL Blue with a 20mW, 488nm, blue laser and 2 PMTs for detection of light scatter and /or fluorescence $55,640 $41,730

CY-S-1031 CyFlow SL Blue with a 20mW, 488nm, blue laser and 1 PMT for detection of light scatter or fluorescence $47,640 $35,730

CyFlow SL includes: • a complete fluidics system with a quartz flow cuvette with a 200µm x 350µm channel cross-section • an optical table with PMTs for forward and side light scatter and up to 3 colors of fluorescence • a solid-state blue (488nm), green (532nm), violet (405nm) or red (635nm) laser • electronics for signal amplification and analog to digital conversion • a desktop or laptop computer running Windows XP and FloMax software for data acquisition and analysis • TVAC sample volume measurement system for determining absolute cell count and cell concentration

CyFlow® SL volumetric flow cytometer with one laser and up to five detectors for scatter and fluorescence

page 1 of 3

page 2 of 3

Catalog # CyFlow SL Green List Price

Discount Price

CY-S-1023 CyFlow SL Green with a green laser and 3 PMTs for detection of light scatter and /or fluorescence $49,640 $37,230

CY-S-1022 CyFlow SL Green with a green laser and 2 PMTs for detection of light scatter and /or fluorescence $41,640 $31,230

CY-S-1021 CyFlow SL Green with a green laser and 1 PMT for detection of light scatter or fluorescence $33,640 $25,230

Catalog # CyFlow SL Violet List Price

Discount Price

CY-S-1043 CyFlow SL Violet with a 100mW , 405nm, violet laser and 3 PMTs for detection of light scatter and/or fluorescence $71,640 $53,730

CY-S-1042 CyFlow SL Violet with a 100mW , 405nm, violet laser and 2 PMTs for detection of light scatter and/or fluorescence $63,640 $47,730

CY-S-1041 CyFlow SL Violet with a 100mW , 405nm, violet laser and 1 PMT for detection of light scatter or fluorescence $55,640 $41,730

Catalog # CyFlow SL Red List Price

Discount Price

CY-S-1013 CyFlow SL Red with a 25mW , 635nm, red laser and 3 PMTs for detection of light scatter and/or fluorescence $49,640 $37,230

CY-S-1012 CyFlow SL Red with a 25mW , 635nm, red laser and 2 PMTs for detection of light scatter and/or fluorescence $41,640 $31,230

CY-S-1011 CyFlow SL Red with a 25mW , 635nm, red laser and 1 PMT for detection of light scatter or fluorescence $33,640 $25,230

CyFlow® SL

Catalog # options List Price

Discount Price

16-01-2000

RobbyWell automated sample loader loads samples from 96-well microtiter plates. (1 minute per well) $34,000 $25,500

Fournier, Michel 14327

1

INTRODUCTION Resubmission, This NSERC/ANR proposal has been submitted to last competition and it was recommended for funding by the panel (1565). All questions and comments raised by the panel members and external experts have been addressed in this improved version. As suggested, particular care was brought to clarify and simplify the text, For instance, a flow chart (Table 2) was added to present milestones and student involvement as well as a Table to summarize the role of members in student training (Table 3). Relationship between markers was also detailed in a Table (1). Relationship with NSERC stated objectives, In this program, we propose to develop a monitoring strategy to assess biological effects of pollution, in combination with climatic changes, through the use of one marine and one freshwater bivalves considered as adequate sentinel species. We will link biological responses with a number of genetic (population level) and physiological (individual) markers to characterize the health status of selected populations. A “Tier approach” will be developed including assays designed to assess 1) impacts on physiological competence, 2) cellular and molecular targets, 3) cellular and molecular mechanisms. The program therefore fits in the research topic “Enhancing Aquatic Ecosystem Services” of the target area on “Environmental Science and Technologies”. It also falls under the 2nd priority “Understanding recent changes in the Arctic and cold region environments” of the new NSERC program on “Climate Changes and Atmospheric Research” and consequently under the priorities of the “International Group of Funding Agencies for Global Changes Research” in which NSERC and French ANR are key members. Rationale and historic context of the international cooperation. Bivalves play an essential role in maintaining the structure and the balance of several marine and freshwater aquatic ecosystems. As sedentary organisms, filtering large volumes of water, they represent good indicator of the health of ecosystems on which they depend and could therefore act as sentinels to follow trend of changes of habitat conditions (climatic changes, pollution, etc.). Several species are used for traditional or economic reasons (gathering or farming) and they are considered as a delicacy to many peoples living along coastal shore. Our knowledge of bivalves and their physiology is therefore essential to the development of aquaculture techniques as well as for assessing the quality of aquatic habitats. These organisms represent also excellent animal models to undertake mechanistic research to elucidate interactions between various physiological systems. Over the past decade, researchers in America, Europe and Africa have worked on the development of physiological endpoints that can serve to assess health status of bivalves. In 2004, under the initiative of Michel Auffret, from Université de Bretagne Occidentale (UBO) from Brest, in France, and Michel Fournier, from INRS-Institut Armand-Frappier in Canada, the first international workshop on bivalves was hold in Brest. The aim of the workshop was to harmonize methodologies for assessing bivalve health. The initial focus of this work was to standardize phagocytosis assay as a measurement of a key natural immune function. This initiative included researchers from Belgium, Canada, England, Finland, France, Germany and Spain. In addition to the harmonization of methods, the participants recommended the creation of an “International Bivalve Network” with the following objectives:

1. To develop health status biomarkers in relevant bivalve species; 2. To validate techniques and establish international standard approaches; 3. To sort into comprehensive tier approach relevant endpoints allowing:

• the assessment of health status and competence of physiological systems; • the monitoring of impacts of changes in habitat conditions; • the elaboration of molecular approaches in support of mechanistic studies;

4. To assess the robustness of these tiered approaches and endpoints using validated laboratory experiments as well as field studies to monitor :

• the effects of contamination in marine and freshwater ecosystems; • the health status of bivalves in aquaculture;


2

• the impact of climate change on bivalve health; Since 2004, seven meetings w e r e hold in Brest, Quebec Rimouski, Bordeaux, Saguenay, Nantes and Mont-Joli in which methodological harmonization and validation have been pursued. Discussions involving more than 50 researchers, primarily from Europe and Canada were also undertaken to select marine and freshwater sites of interest in order to promote a coordinated research effort. Several research programs were also initiated in order to establish a solid foundation on which we develop the present study. Initial studies were funded by ANR in France and b y NSERC Strategic Grant Program to set up genomic approaches in developing g e n o m i c t o o l s i n blue mussels (M. edulis). The currently proposed unique NSERC/ANR collaborative program is based on these consensus and joint research projects since 2004. SECTION 1 Research program 1.1 Research Team 1.1.1 Canadian team. A key strength and a major innovative aspect of this application is the unique interdisciplinary aspect of the research group, which links together experts in immunotoxicology (M. Fournier, (MF); P. Brousseau, (PB)), toxicology of neuroendocrine and reproductive systems (D. Cyr, (DC), F. Gagné, (FG); C. Vaillancourt, (CV)). Each of them represents key scientists in their respective field. They also shared expertise on complementary approaches such as field biology (Y. de Lafontaine, (YL)), controlled laboratory exposures (S. Masson, (SM); R. Vaillancourt, (RV)), biochemical tools (FG, P. Spear, (PS)), cellular (MF, PB, CV), and molecular methods (DC, A. Siah, (AS),S.B. Kilbenge (SBK); Y. St-Pierre, (YSP)) and chemistry (M. Lebeuf, (ML); L. Martel, (LM); M. Kwan (MK); F. Bellemare, (FB)). Moreover, many of them (DC, MF, PB, YL, AS, FG, ML, LM) have collaborations since at least 10 years, as proven by common production ( publications, communications, co-directions of students, [see CVs]). All of them, either co-applicants or collaborators, are members of Centre interinstitutionnel de recherche en écotoxicologie (CIRÉ) as well as the International network on bivalves (EcoBIM). They have all been involved in the proposal preparation. M. Fournier, as PI, will be responsible for the overall coordination of the project, management of the budget and will ensure that the milestones are met. He will also coordinate preparation of reports and communications between team members. 1.1.2 French team. The French multidisciplinary team is composed of researchers from five institutions according to ANR rules. From LEMAR (UBO) in Brest, M. Auffret (MA), PI of this team, will focus on strategy for immunotoxicological approach whereas, D. Moraga (DM), will focus on toxicogenomics and V. Pichereau (VP) on proteomics. From IAE (Université de Reims Champagne Ardenne), S. Betoulle (SB), will be involved in immunotoxicological approach and will assume the coordination of Kerguelen Islands campaigns. A. Bigot (AB) will focus on strategy for gene expression, whereas É. David (ED), scientific leader, will be responsible of gene expression. Finally, A. Geffard (AG) will assume strategy for field sampling and sites selection. From LEMA (Université du Havre), J.M. Danger (JMD) will be involved with molecular aspects. C Minier and F. Le Foll will respectively be involved in data treatment and immunotoxicity. From BioMea (Université de Caen), J.M. Lebel (JML), K.Costil, and A. Serpentini will act as specialists for genomics and reproductive state approaches. From LIEBE (Université de Reims Champagne Ardenne), L. Gamberini (LG) will supervise analysis of digestive lysosomes as well as gametogenesis, C. Cossu-Leguille (CCL) will monitor anti-oxidant enzyme activities in digestive gland, while S. Devin (SD) will be involved in data processing and statistics. Finally, S. Pain-Devin (SPD) will take in charge analysis of MXR and GST responses in digestive gland and /or gills of bivalves. All members are involved in data treatment and comprehensive analysis. A group of 4 students and 15 research professionals will assist these researchers. A great cohesion is existing within this team since they were at the origin of a multi- institutional group named IMOPHYS (impact of environment on mollusk physiology), between 2006 and 2009 and more recently in EXECO (impact of pollutants on coastal


3

environment). These two groups have traditionally key collaborations with Canadian researchers, members of CIRÉ. 1.1.3 Complementarity between Canadian and French teams. In addition of clear collaborations within each national team, we must emphasize that there is a tradition of collaboration between the teams as pointed out in the rationale (see Introduction). For instance, both PI are collaborating since 2004 in several research programs supported by ANR or IFREMER. Moreover, NSERC strategic program established in 2009 [1] puts in collaboration MF, AS and researchers from UBO (MA, DM) and Université du Havre (JMD), all of them being members of the French team submitting the ANR counterpart of the program. French collaborators are also members of CIRÉ. CIRÉ encompasses 150 professors, scientists and managers in Eastern Canada and Europe. There is a well- developed sense of collaboration among the various members allowing further opportunities for students to interact with other toxicologists. We can signal that in the past 5 years, almost 10 PhD students were co-directed by Canadian and French researchers involved in the actual program. These collaborative efforts also include methodological interlaboratory harmonizations. Canadian universities scientists involved in the project have strong training programs in environmental toxicology (INRS, UQAM) and oceanology (UQAR). Furthermore, the toxicology groups at INRS and UQAM recently moved into brand new facilities adding to an already strong research environment. 1.1.4 Complementary perspective between Canadian and French proposals. As described above, the present unique collaborative research program is the result of numerous exchanges. Both proposals share common objectives, research strategies and tasks. Complementarity will also be achieved by the involvement of many graduate students, co-supervised by Canadian and French scientists (each student being registered in both, French and Canadian universities). Similar methods and endpoints will be monitored in selected laboratories from each country. The NSERC proposal differs from the ANR proposal by the greater priority devoted to controlled open flow laboratory exposures (Task 1), to mimic habitat changes conditions (performed only in Canada, with involvement of French students). Because of the well-established expertise by the French team on global analysis (Task 7), this latter task is more developed in the ANR proposal. Implication of Canadians will be through a co-direction of one Post-Doctoral Fellow (PDF). Finally, whereas French scientists will have their own sites of concern along the French coast, Canadian will have their own too. Experiments in the Kerguelen Archipelago will be performed by researchers from both countries, with a special scientific and financial support from the Institut Polaire de France (IPEV). The total cost of the entire research program (NSERC / ANR) is: CAD $ 3 766 306 with the following ventilation: $ 7 3 2 , 2 7 5 requested at ANR; $ 690,366 requested at NSERC; o v e r 1 , 7 M $ provided by partners and collaborators and $ 256 200 provided by IPEV. 1.1.5 Communications between team members. Quarterly meetings between students, staff and collaborators will be held at regional levels. Furthermore, a bi-annual meeting with representatives of our partners will be held to describe progress and to discuss how the research and the developing approaches and technologies could be the best integrated into monitoring program. EcoBIM annual meeting (hold in alternation between France and Canada) will also represent a special occasion to discuss progress of the research with French collaborators. Videoconference and a Forum Discussion Group (webinars) will assure direct communications between all researchers and students, as frequently use in other collaborative programs performed in the past. 1.2 Objectives, tasks and work plan. In the scope of short and mid-term objectives described in the previous section, the working objectives of this strategic proposal are: • To assess the sensitivity of selected health biomarkers and tiered approach for a marine and a freshwater bivalves which react to changes in habitat conditions; • To validate the use of these biomarkers and tiered approach into sites susceptible to become sites of concern for the monitoring of:


4

contaminant effects; health status of bivalves as a food source; impact of climate changes;

To achieve these objectives, the following three-step research strategy has been elaborated:

1.2.1 Step 1: Assessment of habitat changes on physiological competence of bivalves. This first part of this research program has been designed to monitor the impacts of changes of various habitat conditions (water temperature, salinity, pH) on physiological systems. All the work performed by our network in the past decade clearly demonstrates that the immune, neuroendocrine and reproductive systems are influenced by certain abiotic factors such as the position of the bivalves on the beach, the density of organisms in relation to growth, etc.[2,3,4,5,6]. Changes of temperature, salinity and acidification can affect basic physiological parameters of bivalve like respiratory rate, filtration and clearance rate of particles [7,8]. Such perturbations will in turn affect growth and reproduction due to energy requirements in stressful habitats and decreasing energy storage available for gametes development. Studies also reported that changes in salinity and temperature affect neurosecretory function [9] and immune competency [10,11] which can also be impaired by acidification [12,13]. It is therefore imperative to determine the impact of habitat changes and related parameters on the physiology of bivalves. It will be based on health assessment of bivalves using two complementary approaches: (1) from controlled open flow laboratory experiments; (2) from indigenous animals collected at various sites. This second part will also serve to select the most relevant sites to perform the main validation experiment (Task 6). 1.2.1.1 Tasks 1a; 1b: Assessment of biomarkers to habitat changes. This part of the project will be performed with stocks of wild bivalves collected from two reference sites: Baie-de-Mitis and Canal Soulanges for Mytilus edulis and Dreissena polymorpha respectively. Bivalves will be collected at three time points during the year, which represent critical stages in the life cycle of these organisms: winter rest, growth and reproductive periods. The physico-chemical characteristics of water will be recorded at the sampling sites and the maintenance conditions in the laboratories will be adjusted (temperature, salinity, pH, dissolved O2) to reflect field conditions. Animals will be acclimatized in these conditions for one month. Following the acclimatization period (T0), bivalves will then be submitted to gradual changes in habitat conditions, of a single parameter, over a period of two months after which, the animals will remain in this condition for another two months, before testing (T1). A control group consisting of animals held under initial conditions will also be carried out. The selected water temperatures will be as field conditions, 5, 10, and 20°C. For M. edulis only four different water salinities will be also tested: field conditions, 15, 20 and 30 ppm (selected based on known limits of resistance of the species to salinity)[14,15]. For water pH, the actual pH of reference site being 8.0 ± 0.1, this pH will serve as a reference. The following pH, 6.8, 7.2, 7.6 will be tested as result of acidification of seas [16,17]. Experiments with M. edulis will be performed at Aquarium du Québec (Quebec City), while experiments with D. polymorpha will be done at Biodôme (Montreal). During the experiments, animals will be maintained and fed accordingly to previous establish standard procedures. At each time point (T0, T1), 10 animals per group will be studied using selected endpoints described in Tier 1, battery of assays that is designed to screen the impacts on physiological competence (hemocytes count, viability and phagocytosis as detailed in section 1.3). All tissues required to evaluate endpoints of Tier 2 (cellular and molecular targets) and Tier 3 (cellular and molecular mechanisms) will also be collected and stored under optimal conditions for further analysis. Based on results obtained in Task 1a, a companion experiment (Task 1b), in which health status of M. edulis and D. polymorpha will be evaluated using Tiers 1, 2 and 3 following an exposure to the combination of habitat changes (temperature, salinity, pH for M. edulis; temperature, pH for D. polymorpha), which have shown to modulate the physiological competence in each species.


5

Endpoints Tier 1

descriptors for physiological steady-state

Tier 2 descriptors for cellular & molecular targets

Tier 3 genomics & proteomics

General health status and stress

- condition index - growth - scope for growth

- lysosomal destabilization - oxidative stress and antioxidant defence system - cell stress response gene expression - genotoxicity markers

global approach for differential gene and protein expression

Immune status - immunopathology: hemocyte mortality, cell counts - phagocytosis function and related killing mechanisms

- hemocyte hydrolytic enzymes - apoptosis - lipidic composition of hemocyte membrane - immunity gene expression

Reproduction and endocrine status

- gonad weight - gametogenesis stages - vitellin-like contents

Serotonin, dopamine, sexual steroid (estradiol; testosterone, progesterone) level

Metabolism - metabolism gene expression

Pathology - microbial burden - leukemia markers

Table 1 : Tier‐based descriptors approach for experimental and observation tasks. 1.2.1.2 Task 2: Sites selection based on physiological competence. This Task 2 is designed to assess differences in physiological competence of bivalves according to sites location and history. We will therefore apply Tier 1 on bivalves collected on each site at two periods of the year for which reproduction does not interfere with physiological status (early spring and late summer). The selected sites are described in section 1.3. The Tier 1 assays will be performed on 10 animals directly at the collection sites with validated methods adapted for field conditions. At the time of collection, habitat characteristics a n d p h y s i c o -chemical characteristics of water (temperature, salinity, pH, dissolved O2) as well as food availability at each site will be documented. In addition, for the sites retained for Task 6, animals will also be collected to determine genetic diversity as well as incidence of leukemia. Moreover, 1 PhD student (4) will be involved for the work in Kerguelen Archipelago. 1.2.2 Step 2: Ecotoxicological evaluation. The following Tasks are designed to assess the cumulative impact of pollutants on sensitivity of hemocytes collected form organisms subjected to habitat changes (Task 1) or collected from different sites (Task 2). These experiments will make use of in vitro exposures to model toxicants. 1.2.2.1 Task 3: Assessment of sensitivity to in vitro model toxicants exposure. For this work package, we will benefit from the experiments described in Task 1. At each selected time-point, hemocytes will be collected and exposed in a dose-response manner to selected model toxicants (details in section 1.3.4). In order to define the toxicity threshold we intend to assess both viability and phagocytic activity. Because of the design of the experimental protocol in Task 1, this part of the study will allow us to determine the impacts of habitat characteristics on the sensitivity of hemocytes to contaminants following in vitro exposure. 1.2.2.2 Task 4: Assessment of sensitivity of endogenous organisms to in vitro exposure to model toxicants. This work package will benefit from the experiments described in Task 2. At both selected time points, hemocytes will be collected from 10 individuals from each site and exposed in a dose- response manner to model toxicants (details in section 1.3.4). Results will be used for the final selection of sites for field validation experiment (Task 6). This part of the project will be conducted by a research assistant (M. Fortier) and two PDF (M. Gélinas and A. Bruneau). 1.2.2.3 Task 5: Assessing the body burden of pollutants. For each proposed site (see section 1.3), enough bivalves to obtain the necessary mass of tissues, to perform the suite of chemical analyses, will be collected and adequately preserved until analysis. Metal burden will be performed by 1 PhD student (no. 5 {MK/MF/PB}: to be recruited) for M. edulis collected in Kuujjiak area, monitoring of metals in the water column will also be evaluated to define bioaccumulation. The work will also include


6

monitoring of some biomarkers such as viability and phagocytosis. Among chemicals that will be analyzed we can cite: model PAH, and a suite of PCB and PBDE congeners including DBE209. Chemical analyses will be performed according to currently existing methods in each laboratory and performed under the Spirit of the Good Laboratory Practices (see section 1.3.5). In addition, analytical method developments for the analysis of some pollutants and their metabolites will be performed. For one marine and one freshwater site, PhD student no. 6 will also assess the ecotoxicological risk (ERA) by studying the linkage between body burden and presence of pollutants in the exposure media (suspended particulate matter; water column sampled with a diode array detector {SPMD}) and their correlation with affected endpoints. The endocrine disruption potential of exposure media (water column) will also be assessed based on screening reference in vitro tests, in an attempt to integrate the multi toxic chemicals exposure in the ERA. 1.2.3 Step 3: Field validation. This final experimental part of the research program has been designed to evaluate and validate the sensitivity of selected markers under field conditions in both freshwater and marine ecosystems. The analysis of the results will enable us to establish a validated toolkit that can subsequently be used for sentinel species in reference sites. This approach and the methods used represent the basis for future monitoring of combined effects associated with habitat changes due to climate changes and anthropogenic activities including pollution. 1.2.3.1 Task 6: Field validation. This part of the project will involve an annual follow-up of stocks of wild bivalves collected from both marine and freshwater selected sites. At the time of sampling, habitat characteristics, physico-chemical characteristics of water (temperature, salinity, pH, dissolved O2) as well as food availability of each site will be documented. This field follow-up will be performed hopefully through the year, accordingly to accessibility to organisms. Indigenous organisms will be collected monthly and used to assess endpoints of Tiers 1, 2 and 3. In addition, sensitivity of hemocytes to additive stress to exposure to pollution will also be studied using in vitro exposures with model toxicants (see section 1.3.4). Comparisons of dose-response curves will enable the determination of the impact of habitat condition changes on the sensitivity to chemicals. 1.2.3.2 Task 7: Global analysis of data. Only a full set of biomarkers will allow the ecotoxicological approach to identify biological effects of a complex contamination scenario in a complex environment [18]. The main two objectives of this task through inter-laboratory approach are: i) identification and classification of the most relevant markers (individual and population levels) for the characterization of temporal evolution of the health status of populations under a combined water contamination and habitat changes stress; ii) assessment of sensitivity of species in terms of vulnerability to targeted anthropogenic stressors. For that purpose, multivariate statistical tests will be applied according to methods proposed in the ECOMAN project [18].

The ultimate objective of IPOC is to propose an adequate long-term surveillance program in suitable sites. To achieve this goal, we propose an integrated processing of data where selected biomarkers of exposure and effects would be combined to numerical values. In the last decade, various monitoring programs around marine hot-spots of pollution have provided ecotoxicologists with a substantial volume of data allowing to develop a set of integrated indices [19]. We intend here to apply and compare three methods already validated in marine mussels:


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1. Integrated Biomarker Response (IBR) [20], which originally included biochemical biomarkers of exposure. Since its elaboration, other authors have included biomarkers of effects and of general health [19] ; 2. Immunotoxicological index (II) [21] which focus on various cellular and biochemical responses detected in immune system of mollusks exposed to environmental stressors; 3. Biomarker Response Index (BRI) [22] which was developed to improve risk assessment of chemicals in Europe. By including this approach in various scenarii linked to global change, IPOC could provide relevant issues for environmental management in North-Atlantic region.

Months Task 0-6 6-12 12-18 18-24 24-30 30-36

1a M.Sc 1,2,3 PhD.1,2,3 M.Sc 1,2,3 PhD.1,2,3 M.Sc 1,2,3 PhD.1,2,3 1b M.Sc 1,2,3 PhD.1,2,3 M.Sc 1,2,3 PhD.1,2,3 2 M.Sc. 4,5 PhD. 4

PDF 1,2 M.Sc. 4,5 PhD. 4 PDF 1.2

M.Sc. 4,5 PhD. 4 PDF 1.2

3 PhD 2 PhD 2 PhD 2 PhD 2 PhD 2 4 PhD 1,2 PhD 1,2 PhD 1,2 5 MSc 6 PhD 5,6 MSc 6 PhD 5,6 MSc 6 PhD 5,6 MSc 6 PhD 5,6 PhD 5,6 6 MSc 2,3 PhD 1,2,3 MSc 2,3 PhD

1,2,3 MSc 2,3 PhD 1,2,3

7 SPD3 SPD3 Table 2. Flow chart of the different tasks and students involvement (experimental part, writing of thesis not considered). 1.3 Research approach, endpoints, methods, sites of concern and toxicants of interest 1.3.1 Endpoints. It is important at this step to review endpoints that can be monitored in bivalves and could be considered in the integrated approach we are proposing to develop. Special care has been given to endpoints relevant to physiological systems known to be the target of endocrine disruption. These endpoints have been monitored in bivalves capable of living in marine, brackish as well as in freshwater. We therefore will have a model that could be applied for marine and estuarine environments but also freshwater ecosystems such as rivers and lakes. In order to rationalize the use of these endpoints in the most efficient way, we propose to regroup them, using tier approaches, into three groups of endpoints: Tier 1, Screening of physiological competence, will regroup general endpoints on growth and welfare and selected parameters that can pinpoint the competence of targeted physiological systems; Tier 2, Identification of cellular and molecular targets, will regroup endpoints at cellular and molecular levels explaining impacts revealed by Tier 1; Tier 3, Elucidation of cellular and molecular mechanism, will focus on biochemical and genomic assessment of cellular and molecular mechanisms behind adverse effects. A similar Tier approach has been developed to assess side effects of drugs and has made a consensus among regulatory agencies [23,24,25,26,27]. 1.3.1.1 Endpoints assessing sites characteristics. In addition to traditional characteristics of the environment (pH, temperature, salinity, dissolved O2) we will also monitor more appropriately habitat characteristics such as sediment granularity, distance of clam beds from the shore, number of empty shells (predation), particulate material, chlorophyll a). Moreover, LuminoTox PEC and SAPS portable Test Kit biosensors, develop by Lab_Bell Inc., which indicate the presence of toxic chemicals in water, will also be used. Moreover, chemical analysis will be done for metals and organic chemicals. 1.3.1.2 Endpoints assessing general health status. Based on our experience, the most important endpoints to be considered as significant parameters to evaluate health status are: age, size, weight, oxygen consumption and clearance filtration rate (suspended particulate matters) [5,6,28]. All those endpoints have been shown to impact on heterogeneity of physiological responses. As far as possible, collection of organisms and their subsequent selection for testing will be based upon those parameters. Therefore, these parameters are included in Tier 1 and some parameters will allow also the determination of growth (Tasks 1a, 1b, 2). 1.3.1.3 Endpoints assessing immune system. The outstanding use of the immune system in the context of aquatic ecotoxicology relies on its exquisite sensitivity, intimately linked to the sophistication of its


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structure. Immune system in bivalves, like for all other organisms, are permanently exposed to antigenic stimulations. While in vertebrates, immune system includes acquired adaptive immunity and innate or natural immunity in invertebrates, protection of the organism is assured solely by natural immunity [29]. In bivalve, natural immunity is composed of cellular components (mostly hemocytes) and humoral components consisting of molecules circulating in hemolymph. Hemocytes, distributed in all tissues, represent the main immune cell population. They constitute a morphologically heterogeneous population in several species such as the scallop and the mussel. Hemocytes are capable of phagocytosis of foreign material. They can produce reactive oxygen species (i.e. peroxide) and nitric oxide [30]. They can also secrete antimicrobial substances [29,30,31]. Indeed, disruption of the delicate balance in the regulation of these responses could lead to various unintended responses. Among them, we can mention immunosuppression and immunostimulation. In this context, sensitivity to xenobiotics and other environmental stressors has particularly been studied by our group using multiple approaches (in vitro, in vivo controlled exposure; mesocosms, naturally exposed animals, caged animals) [30]. Immune system was found sensitive to a wide variety of chemicals such as metals following in vitro [32,33,34] and in vivo exposures [33,35], to organothins [36,37], to organic pollutants trapped in marine sediments [38] as well as PAH [39,40], to nanoparticles [41,42] and to municipal sewages using caged bivalves [43,44,45,46,47] or with animals naturally exposed [47]. In a multispecies research program that we undertook (over 30 species), the sensitivity of the immune system to metals, was monitored through a dose – effect manner and bivalves were found among the three most sensitive species [48,49]. We were therefore able to demonstrate the potential utility of bivalves in ecotoxicology [21,29,48,50,51]. Since the last few years, there is a growing body of evidence that immune system of bivalves is also sensitive to environmental stressors, other than chemicals, such as temperature shift, radiation, etc., suggesting that climate changes may represent an important threat to immune competence and resistance to infections in bivalves [2,4,5,6,52,53,54,55]. Affecting immune competence by decreasing resistance to infections can be of first importance, especially in stressed populations. Based on these considerations and observations we considered that hemocytes count and viability and phagocytosis are essential endpoints of Tier 1. All other endpoints will be part of Tier 2, whereas genomic approaches will be part of Tier 3. This last Tier is particularly relevant, since genomic tools were developed to detect the expression of 15 000 genes in hemocytes of M. edulis, during a bacterial challenge, under an NSERC strategic grant [1]. Methodological aspects. All methods have been developed and validated in our laboratories under the “Spirit of Good Laboratory Practice”. They have also been the object of many publications and books [28,30,48,49,56,57]. 1.3.1.4 Endpoints assessing neuroendocrine and reproductive systems. Contaminants alter the neuroendocrine regulation of reproductive system thus affecting sexual maturation and gonadal development. These impairments are closely linked to delays or dysfunctions of the reproductive process leading to decrease in targeted population. Any factor that can alter reproductive processes could then be suspected to be harmful to the ecosystem. Members of the team have shown that chronic exposure to sub-lethal levels of contaminants in the wild bivalves affect the sexual maturation despite the capacity of gonads to store energetic reserves, demonstrating impairment at the gonadal level [58,59,60]. This disruption was associated with a decrease in vitellogenin levels [61] a molecule essential for the development of oocytes, whose synthesis in several bivalves is under the control of steroids (e.g. estrogens). Bivalves have been used as a model for neuroactivity for several decades. Monoamines (serotonin, dopamine), which are good markers of neurotoxicity, are important mediators of gamete maturation and spawning [62]. The latter is activated by serotonin and prostaglandins released by the induction of arachidonic acid cyclooxygenase or COX. Moreover, sex differentiation and gametogenesis represent critical steps in the reproductive process and are subject to hormonal control by serotonin, dopamine and steroids such as 17β-estradiol and testosterone [63].The measurement of gonadal dopamine and serotonin was used to assess the stages of gametogenesis from early


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vitellogenesis to spawning [64,65]. Methodological aspects. General assessment of reproductive health (Tasks 1, 2 and 6) will be evaluate in Tier 1 by histological analysis of the gonad (D. polymorpha and M. edulis) and mantle (M. edulis) to determine the development stage of gametes, GSI determination, and vitellin-like content. Neuroendocrine characterization of gonad tissues will be further assessed (Tasks 1b, 6) in Tier 2 by measuring the following parameters: serotonin and dopamine 1) production, 2) transport and 3) degradation (MOA activity) respectively by LC/MS/MS, RIA transport assays and enzymatic assay (COX and monoamine oxidase activities). All methods are routinely used in our laboratories. 1.3.1.5 Endpoints assessing oxidative stress and antioxidant defense systems. Reactive oxygen species or ROS (e.g. •OH, O2

-, O2 , H2O2) are generated in all aerobic cells and are counteract by both indogenous antioxidants (e.g. vitamin E, carotenoids, reduced GSH) and antioxidant enzymes (e.g. superoxide dismutase, catalase, GSH-peroxidase). In M. edulis subjected to a transient high temperature, lipid peroxidation rate was increased in the digestive gland while antioxidants (reduced GSH, vitamin E) decreased [66]. However, in this tissue, the balance between pro- and antioxidant is potentially altered by many confounding factors including seasonal variation, reproductive cycles and changes in food quality [67]. Antioxidant enzymes also act as protective enzymes in immune cells and hemolymphe of bivalves in response to ROS production during the oxidative burst [68,69]. ROS production of hemocytes is also expected to increase with increasing habitat temperature as demonstrated in hemocytes from oysters [10,70]. Cold-adapted ectotherms generally are susceptible to oxidative stress due to the greater oxygen saturation in cold water and the greater proportion of polyunsaturated lipids required to maintain cell membrane fluidity, and presumably as a consequence, their requirement for vitamin E is known to be increased [71]. However, organohalogen contaminants have so far been shown to decrease vitamin E in fish and birds [72,73], while β-carotene does not appear to be altered [74,75,76]. Such effects have not been investigated previously in bivalves. The vertebrate literature (in the absence of research into bivalves), proves that adequate vitamin A (including carotenoid precursors) and vitamin E are essential to a competent immune system and related disease resistance [71] in addition to maintaining reproduction, embryo development, growth and other physiological processes [77]. Bivalve tissues contain a broad spectrum of carotenoids and xanthophylls as potential antioxidants, although their physiological role is only partly understood. In oyster hemocytes, it was shown that mitochondrial respiratory chain is strongly involved in the oxidative activity of hemocytes (ROS and RNS) [78]. Since restructuring of membrane composition (lipids and fatty acids) may play an important role on adjustments of mitochondria capacity in response to thermal variations[79]. ROS production in hemocytes of bivalves subjected to thermal variations should be investigated in close correlation to their mitochondria capacity. With respect to oxidative stress and antioxidant enzymes, bivalves exposed to various types of environmental contaminants have increased lipid peroxidation rates and commensurate decreased enzyme activities [3,80,81]. Methodological aspects. Lipid peroxidation (thiobarbituric acid method) and reduced GSH (fluorometric assay) will be analyzed in crude homogenates of the digestive gland. Antioxidant enzymes (superoxide dismutase, catalase, GSH-peroxidase) will be assessed by standard methods in the post-mitochondrial fraction of the digestive gland. The antioxidants vitamin E (α- tocopherol), major carotenoids and major xanthophylls will be extracted from the digestive gland by homogenization with organic solvents and quantified by reverse-phase HPLC (method under development in our laboratory). ROS production in hemocytes of mussels will also be determined on fresh cells by spectrophotometry [21] while antioxidant enzymes (SOD, CAT, GPx) will be further determined by spectrophotometry on hemocytes and hemolymphe properly preserved [82]. Mitochondria will also be extracted from preserved hemocytes to determine their capacity (ETS, CCO, CS) by standard methods used in our laboratories [83]. All these parameters will be part of Tier 2.


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1.3.1.6 Endpoints assessing mechanistic aspects. In order to understand the cellular and molecular effects of environmental contaminants on bivalves it is essential to understand low-level effects on intracellular signaling pathways involved in biochemical and physiological processes. We propose to use a genomics approach to first identify targeted pathways and to subsequently assess the consequences of alterations of down-regulated pathways in vitro. To identify specific genes/pathways whose expression is altered by treatment, we will examine gene expression profiles on RNA isolated from hemocytes. In a previous study, a cDNA library of hemocytes (M. edulis) leading to the annotation of more than 15,000 unique ESTs have been constructed and sequenced using the 454 Roche technology [84].These sequences will serve as reference transcriptome for the analysis of gene expression levels in the different conditions of exposure by using NGS technologies. Focus will be made on profiling gene expression in hemocytes submitted to a Tier combination of exposure to contaminants and/or abnormal temperature mimicking global changes. Quantitative PCR and NGS of transcriptomes of hemocytes will be performed in order to compare gene expression patterns and to extract single polymorphism data reflecting the “population status” of mussels. It will also be of particular interest to analyze these field data with those obtained with our in vitro experiments to eventually address the “transcriptomic state” of animals in the wild. Methodological aspects. Bioinformatic analysis will be performed by using the facilities offered at the GenomeQuest Server. Following quantitative PCR and NGS, the data will then be entered into the pathways analysis program (IPA, Ingenuity Software), to predict altered signalling pathways. We have recently used this approach to identify altered signalling pathways in fathead minnows exposed to municipal wastewater effluent. Since there are no databases for bivalve signalling pathways, we will use mammalian databases to identify altered signalling pathways. These analyses are based on the assumption that biochemical signalling pathways have remained largely conserved throughout evolution. There is no data now to support or refute this assumption. Therefore, we intend to test the consequences of alterations in these pathways by specifically inhibiting predicted pathways using either siRNA or morpholino antisense oligonucleotides (Gene Tools, LLC) and target specific genes implicated in the initiation of these signalling pathways. This approach will not only provide information regarding alterations in cellular signalling pathways following exposure, but will also enable us to demonstrate the consequences of these alterations on hemocyte physiology. 1.3.1.7 Endpoints assessing pathologies. Shellfish in all environments become stressed, as a result of environmental contaminant exposures that may translate into direct toxic action on tissues, alterations in homeostatic mechanisms or into a higher prevalence of disease such as haemic neoplasia. Growing evidence suggests that heavily contaminated environments correlate with haemic neoplasias, redefined as leukemias, in bivalves [85,86,88]. Among mollusks, mussels (M. edulis and M. galloprovincialis), and clams (M. arenaria and Mercenaria spp.) have the highest number of reported cases of leukemia [89,90]. Laboratory studies on soft shell clams [91,92] and blue mussels [93]

confirmed the progressive and fatal nature of the disease over a period of 3 to 6 months. Methodological aspects. Disseminated neoplasia is commonly recognized as tetraploid disorder [94,95]. This disorder is related to the sequestration of p53 protein in the cytoplasm [96], high levels of p53 gene expression [97] and mutations of this gene [98]. It is well accepted that specific genotypes of p53, a key factor in the cell cycle regulation, could be used as a checkpoint for when hemocytes become neoplastic. In our research, we have utilized a combination of microscopic examinations and monoclonal antibodies to document the induction of leukemia in M. edulis. These techniques are complemented with molecular approaches which aimed at identifying, by DNA sequencing, specific variations in the coding region of p53 associated with the development of haemic neoplasia. 1.3.2 Species selected. Mytilus edulis. Blue mussel, Mytilus, is a common bivalve forming large populations on coastal, natural and artificial hard substrates, including rocky shores and concrete wharfs in harbors. This mollusk has a particularly wide geographic distribution since it is found in the Atlantic Ocean, the North and Baltic seas and M. edulis desolationis is present in Kerguelen


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Archipelago. As a filter feeder, it has been widely used as sentinel species for the survey of chemical quality of waters and in ecotoxicological studies. The blue mussel is able to survive in artificial conditions and is then a suitable species for experimental studies to chemical contaminant exposure and habitat changes. Because of its ecological importance as well as its economic value, extensive research has been performed on its physiology [29,30,50]. Dreissena polymorpha. The ubiquitous nature of zebra mussels in Canada and Europe stimulated the interest to use it as potential sentinels in aquatic ecosystems, for pollution stress like municipal wastewaters [99], endocrine disruptors and bioenergetics studies [100,101,102,103]. Many experiments on the ecophysiological behaviour of this species were performed to understand its invasive properties. 1.3.3 Sites of concerns (for more informations refer to the section “environmental impacts” of the main document) 1.3.3.1 Marine sites Baie-de-Mitis. This reference site has been the object of an intensive scrutiny by our group in last two decades. The bay is well preserved from any industrial activity and the main group of mussels is not accessible to public. Laboratory facilities, provided by Parc de la Rivière Mitis are available for the research team, directly on the shore. Saguenay River. The Saguenay fjord has suffered of an important polyaromatic hydrocarbon (PAH) and metal contamination since the 1950’s. The fjord was also part of a network devoted to monitor bacterial quality of coastal zones in Eastern Quebec, under the supervision of Environment Canada. This site is under investigation by a research group involving many of the present collaborators since 1995. Work performed with Mya arenaria has been the object of numerous publications [5,6]. In the last three years, M. edulis has also been the object of research, especially at the mouth of the Saguenay River where salinity is at limit of habitat for the blue mussels.Baie-des-Chaleurs. The Baie des Chaleurs watershed is located in Northeastern New Brunswick and the Gaspésie peninsula in Quebec. It is a rich and diverse fresh and marine ecosystem with important river systems. It is one of the most important estuaries draining into the Gulf of Saint Lawrence. Baie des Chaleurs region is dealing with important erosion problems and sea level rise due to climate change. In addition, the Baie des Chaleurs region is now dealing with closing industries and a legacy of industrial contaminants [104] which can re-circulated to the aquatic environment by erosion of the coastline. Kuujjuak area. The coastline of the Nunavik Region of Northern Quebec encompasses the East Hudson Bay coast, the south coast of the Hudson Strait and the Ungava Bay. The 14 Inuit communities along this coast line all have accessible blue mussel beds where Inuit harvest the shellfish in both winter and summer as part of their country food diet. Largely, all the mussel beds are located in a remote and pristine sub-arctic environment and mussel beds near the communities of Kangiqsujuaq, Kangiqsualujjuaq and Kuujjuaq are suitable uncontaminated sampling sites for this study. At the opposite, the mussels bed in Deception Bay near the community of Salluit, where active nickel mining has been operational for the last three decades at the nearby Raglan mine organisms collected at this site have levated levels of various metals including nickel, chromium, cadmium and zinc detected in the soft tissues. Climate change, which is happening very rapidly in the North, has an important implication on water quality as well as a potential direct influence on the physiology of blue mussels. Prince Edward Island (PEI). Although PEI is the smallest province in Canada, it ranks first in terms of potato production. This agriculture requires the use of high quantities of agrichemicals such as pesticides and nitrate-based fertilizers (application rates estimated at 220 kg/ha/yr. of fertilizer (Government of PEI 2008). It was evaluated that PEI is the province with the most intensive use of pesticides in Canada, with total pesticide sales in 2000 of over one million kilogram [105]. The contaminated river waters flow out into the sea surrounding the Island, and then expose marine invertebrates inhabiting the intertidal zone to the contaminants. In addition, reduced flushing allows accumulation of nutrients thus enabling a high proliferation of macroalgae, which results in a general decrease in the oxygenation rates of seawater in particular under high temperature conditions. Consequently, high prevalence of pathologies was recorded among aquatic


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invertebrates in the area surrounding the Island. The main example is the development of disseminated neoplasia diagnosed in bivalve mollusks present in North River and Barbara Weit. An intensive monitoring program has been put in place since 2005 to follow health of soft-shell clams. [95].Kerguelen Archipelago. The Kerguelen Archipelago, part of the French Austral and Antarctic Territories, is located at the northern limit of the Southern Ocean. The Polar front runs in the North of archipelago. Moreover, the circumpolar stream arrives in the Western coasts. The associated important water stream is divided in two parts creating distinct hydrological areas in Kerguelen's coastal zone. Such hydrological differences between West and East are also related to major freshwater discharges by rivers in Northern and Eastern part of archipelago Moreover, water temperature is approximately +6°C in North and +4.5°C for the Southern coasts of Kerguelen Islands. In addition of such physicochemical characteristics, high cadmium accumulations were observed in several marine organisms of the Southern Ocean [106,107]. A recent analysis of M. edulis desolationis specimens confirmed this Cd contamination (Geffard, unpublished data). Besides these results on cadmium, Kerguelen's aquatic organisms are contaminated by POPs [108,109] in relation to atmospheric transports. Thus, by its strategically geographical position, Kerguelen mussel populations may constitute major sentinels of the effects of global change on aquatic ecosystems. 1.3.3.2 Freshwater sites. Lake Erie (Great Lakes) where zebra mussels were first reported in late 1980’s. This site is a lacustrine habitat where temperature and other environmental conditions (pH, calcium levels, alkalinity) are optimal for zebra mussel growth and reproduction. Specimens will be collected from sites subjected to minimal contamination influence. Canal Soulanges in the St. Lawrence River is a relatively pristine site with very dense populations of zebra mussels. Located upstream of Montreal Island, this site offers riverine conditions, typically observed in the St. Lawrence River, in terms of temperature and other basic water quality parameters. It will therefore represents our reference site for D. polymorpha. Montreal Harbor is the most important harbor in Eastern Canada and represents a highly perturbed and stressed environment with high levels of contamination, both in sediments and in biota living in the vicinity of the harbor. The harbor also receives wastewater overflow from municipal effluents. Zebra mussels are found in high abundance along the harbor piers and thus are exposed to various contamination stressors. The harbor waters are typically representative of the St. Lawrence River in terms of temperature regime, pH, alkalinity and calcium levels and a relatively high speed current which may provide lesser growth conditions to zebra mussels. Richelieu River is a tributary of St. Lawrence River and was recently invaded by zebra mussel coming from Lake Champlain [102]. The river is characterized by temperature regime similar to the St. Lawrence River but by low calcium levels (15-18 mg/L) which may become limiting to mussels shell growth. Specimens will be collected at two sites in the upper Richelieu River (St-Paul de l’Île-aux-Noix and Chambly). The St-Paul site is located upstream of major industrial cities along the Richelieu River and will represent less contaminated conditions, while the Chambly site, located downstream of urban input, will provide zebra mussels having been exposed to multiple sources and high levels of contamination. Quebec City represents actually the northern limit of zebra mussel distribution in the St. Lawrence River and in North America. This site located in the fluvial estuary is characterized by more severe growing conditions for zebra mussels, due to high-speed flow and tidal influence. Water temperature is on average 2oC lower than that measured in the St. Lawrence River, thus reducing the growth season for zebra mussels. Due to high flow, rapid water exchange and high turbidity of waters, feeding of zebra mussels is considered being not optimal at that site. Moreover, climate change scenarios predict that Quebec City waters should have major impacts, because of temperature increase and possible salinity increased due to reduced flow rate (river discharge) and saltwater intrusion from estuary. Effects on zebra mussels populations at this site are difficult to predict.


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1.3.4 Reference compounds. In order to assess the sensitivity of cells to toxicants as described above (see sections: 1.2.2.1, 1.2.2.2) we have selected five reference substances. For the five chemical selected, our team has studied in details their toxicity. Consequently, we are familiar with their utilization and we know their toxicity on bivalve’s cells. The first one, representative of metals, will be cadmium, which is a well-documented contaminant in several sites selected especially in Kujjuuak, St.Lawrence River, as well as in Baie des Chaleurs [104]. The second one, representative of nanoparticles, will be Quantum Dots with cadmium- tellurium for which, direct toxicity as well as mechanism of action have been studied in bivalves [41,42]. The third and fourth ones are representative of phytotoxins. Microcystin-LR and saxitoxin will be used for D. polymorpha and M. edulis respectively. Two other substances, ethinylestradiol as a representative of endocrine disruptor and benzo[a]pyrene as a representative of PAH will be included when enough cells will be obtained. For all these substances, in vitro exposures will consist of dose-response studies and the effects on parameters tested will be compared using the dose inducing 50% of suppression. 1.3.5 Quality control. Since the beginning of the work undertaken with bivalves, because of the involvement of many laboratories located in various sites, a major effort has been provided to successfully validate and transfert methodologies between different laboratories in order to standardize the tests, to get reproducible results. For endpoints, which require fresh material, all experiments will be conducted by respective laboratories under the Spirit of the Good Laboratory Practices. For endpoints that can be measured from frozen or fixed material, all the samples will be shipped to one reference laboratory to avoid any bias resulting from technical differences that may occur between laboratories. Moreover, tissues used for the monitoring of other endpoints will be preserved in case further analyses are required. SECTION 2 Training of graduate students. The proposed study is particularly well suited for training graduate students in environmental toxicology as well as in reproductive, cellular and molecular biology and ecology. The multidisciplinary nature and the multiple affiliations (academia, governmental, industrial) of the scientists involved in this research provide a unique opportunity for interactions with students in complementary research fields. We must not forget the positive aspect for students to be confronted to the French and Canadian way of thinking. Researchers (co-applicants as well as collaborators) involved in the project will work closely together on all aspects of the study, and graduate students will be co-supervised by at least two members of the team, in case of M.Sc and by three members for PhD. All of the investigators in this study strongly believe that graduate students should present their data at national and international meetings in order to acquire experience and initiate their scientific careers. We would therefore encourage students to submit and present their data at meetings such as the Society of Toxicology of Canada, the Society of Environmental Toxicology and Chemistry (SETAC), the Society of Toxicology (USA) and EcoBIM annual meeting. Students will be strongly encouraged to publish their results in scientific peer-reviewed journals. Efforts have been put forward to provide students with a Forum Discussion Group on the web (webinar), which will facilitate greatly exchanges between students and researchers.

M.Sc. Name Directors Ph.D. Name Directors 1 Lauriol, E. Vaillancourt, C., Gagné F. 1 Fraser, M. Vaillancourt, C., Gagné F., Cyr, D. 2 To be recruted Masson S., Vaillancourt, C. 2 Adam, C. Fournier, M., Cyr, D., Siah, A. 3 To be recruted Vaillancourt, R., Brouseau, P. 3 To be recruted Spear, P., Siah, A., Kibenge, F. 4 To be recruted La Fontaine, Y., Brousseau, P. 4 Casa, F. St-Pierre, Y. Betoulle, S. Fournier, M. 5 To be recruted Fournier, M., Brousseau, P. 5 To be recruted Kwan, M., Brousseau, P., Fournier, M. 6 To be recruted Lebeuf, M., Martel, L. 6 To be recruted Lebeuf, M., Fournier, M., Martel, L. 7 LeQuernic. A. Betoulle, S., Fournier, M., Brousseau, P. PDF 1. Gélinas, M. 2. Bruneau. A., 3. to be recruted 8 To be recruted Auffret, M., Danger, JM, Cyr, D.

Table 3 Student training (French student or director are in italics).


14

SECTION 3 Interactions with supporting organizations, intellectual property. The goal of this study is to generate novel approaches and tools that can be used to predict the combined effects of pollution and climate changes on marine and freshwater aquatic ecosystems. It will also pinpoint sites of concern particularly suitable to serve as benchmark sites. The development of a grid of markers, included in a tier approach, will serve ecotoxicologists and other scientists who study a variety of aspects of aquatic species biology. The partners in this project, such as Environment Canada and Fisheries and Oceans Canada, also represent end-users of the data, which can be applied towards risk assessment as well as for testing the consequences of pollutants and changes in habitats, related to climatic changes, on aquatic organisms in the St. Lawrence Basin. There is no issue about intellectual property in the present proposal because we aim to publish all the data obtained. SECTION 4 Deliverables. Regulating classical and emerging substances and assessing the toxicological risk of exposure to them of wildlife species and human, in combination with climatic changes is quite problematic largely due to a lack of knowledge. The research is undertaken to provide numerous deliverables: At the national level, deliverables will be: 1. Development of a diagnostic kit to assess health status of bivalves, such as blue mussels, a commercially important species; 2. Identification of suitable sites, which can serve as operational base for scientific long-term surveillance programs; 3. Production of comprehensive data, based on ecophysiological and ecotoxicological indices to provide useful quantitative informations to be used in global environmental analyses by scientists, fisheries and environmental managers 4. Formation of young scientists ready to contribute to Canadian society in the future; 5. Formation of highly trained personnel; The deliverables generated will benefit to the Canadian regulatory agencies (e.g., Environment Canada or Fisheries and Oceans) urged to set regulations/guidelines in order to protect wildlife and humans. Moreover, since 1990, shellfish farming represents a significant input to the economy of Atlantic Canada, especially in rural areas. The production and commercialization of mollusks and crustaceans are by far, the most lucrative activities of maritime fisheries. Shellfish farming in Quebec (mussels and scallops) includes 27 plants, approximately 150 annual and seasonal employees and represents an approximate value of $ 1 M (Statistic Canada, 2006, 2009). In the Maritimes region (NB, PEI, NS), commercial production of mussels increased in value from $4.3 to $26.7 million dollars from 1991 to 2009 (Statistics Canada, 2006, 2009). Among the obstacles, which can impede the growth of this industry we find issues associated with sites carrying capacity as well as problems associated with the health of organisms (massive mortality, bacterial and parasitic infections, exposure to toxic substances, reduction in product quality, post harvest). At the international level, deliverables will be: 1. Production of a comparative analysis of data generated in France and in Canada; 2. Recognition of the quality of Canadian research through transfer of knowledge to the International Council for the Exploration of the Sea (ICES) and OSPAR, Commission for the Protection of the Marine Environment of the North-East Atlantic in order to help those implementing guidelines. Members of these organizations are involved in this NSERC / ANR proposal. 3. Increment in methodological expertise of partnership will be further shared through joinedWorkshops. Harmonization and intercalibration of investigative methods applied to sentinel‐species has always been a major concern in our group. This was achieved through EcoBIM workshops held since 2005 for the development and validation of nondestructive immunological methods by flow cytometry. A comparable approach will be included in IPOC for biomarkers based on isolated cell analysis.

Parameters Methodologies Tissues Development stage Laboratories Person in charge Phagocytosis Flow cytometry Hemocytes Published LEMAR M. Auffret ROS Flow cytometry Hemocytes Published INRS-IAF M. Fournier MXR Flow cytometry Hemocytes Development needed LEMA C. Minier

Table 4 : Endpoints to be shared within the international consortium through thematically workshops.


1

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3

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Titre : Chaire de recherche du Canada en Immunotoxicologie de l’environnement Organisme(s) : Chaire de recherche du Canada

Chercheur(s) / Michel Fournier, INRS-Institut Armand-Frappier

Montant / Durée (en mois) : $1,400,000, 2010-2017

Résumé :

Le passage à l'ère industrielle s'est traduit par une augmentation phénoménale de la quantité et de

la diversité des produits de synthèse introduits dans l'environnement suite aux activités humaines.

La présence de ces substances peut constituer une menace sérieuse pour la santé humaine et pour

celle des écosystèmes. En effet, un consensus général se dégage parmi les intervenants en santé

environnementale: l'exposition à certains composés chimiques, pouvant agir sur les systèmes

endocrinien, reproducteur, nerveux et immunitaire des individus, peuvent provoquer des effets

quantifiables néfastes au niveau des populations humaines et de la faune.

A la lumière de ces observations, les objectifs du présent programme sont: 1) déterminer les

relations de cause-effets entre l’exposition aux mélanges de contaminants présents dans le

poisson et les problèmes de santé humaine tels qu’identifiés par les études épidémiologiques, afin

de permettre le développement d’outils efficaces de prévention, (normes d’exposition, table de

consommation, règlements, etc.) ; 2) comprendre les mécanismes cellulaires et moléculaires de

toxicité de ces substances afin, d’une part, d’établir des outils d’évaluation du potentiel

modulateur endocrinien de ces composés avant homologation, et d’autre part, d’identifier des

bio-marqueurs applicables au suivi des populations humaines (catastrophe écologique, évaluation

du succès des actions de prévention, etc.) ; 3) bien documenter la question de la plus haute

sensibilité à l’exposition aux toxiques telle que démontrée chez les organismes en développement

et en déterminer les causes.

Liens avec la présente demande : Le choix des substances retenues dans la présente demande

est basé sur les travaux effectués dans le cadre de la Chaire de recherche du Canada.

Titre : Influence du stade de développement sur l'immunocompétence et la sensibilité aux

infections chez les mollusques bivalves.

Organisme(s) : CRSNG Stratégiques Chercheur(s) / Michel Fournier, Gauthier-Clerc Sophie, Pellerin Jocelyne, Lemarchand Karine, Siah

Ahmed, Auffret Michel, Danger Jean-Michel, Moraga Dario.

Montant / Durée (en mois) : $470,000, 2006-2011

Résumé : Le projet a pour principal objectif de caractériser les défenses immunitaires des

mollusques bivalves durant leur développement larvaire et au stade adulte, et de mieux

comprendre leur efficacité en cas d’infection bactérienne. Les connaissances dans ce champs de

recherche sont encore parcellaires et les retombées de ce projet seront particulièrement

novatrices. La moule a été choisie comme « modèle biologique ». Elle est aussi le bivalve le plus

cultivé au Canada. Les ventes du secteur mytilicole présentent près de 32.8 millions de dollars.

Ce programme de recherche implique sept chercheurs de deux provinces de l’est du Canada et

trois instituts de recherche et contribuera à la formation de six personnes hautement qualifiées. Il

permettra également le développement de techniques et d’outils moléculaires de pointes. Un

transfert technologique depuis des laboratoires de recherche français hautement spécialisés en

génétique aura également lieu grâce à la collaboration trois chercheurs français. Deux approches

seront utilisées pour étudier les défenses immunitaires des différents stades larvaires et des

adultes ainsi que leur réactivité pour faire face à une infection. La première caractérisera les

différents composantes cellulaires et humorales de la capacité immunitaire des larves et des

adultes et la seconde se concentrera sur l’expression des gènes impliqués dans l’immunité. Les

outils de diagnostic de la santé des bivalves développés dans ce programme de recherche

permettront à l’industrie conchylicole d’optimiser la productivité des élevages en détectant de

manière précoce une dégradation de la santé des moules à cause de maladies ou de pratiques

zootechniques trop stressantes. Les connaissances acquises sur la capacité des larves de bivalves

à résister aux infections sera aussi un atout majeure pour développer des approches novatrices de

cultures larvaires capables d’optimiser la performance d’écloseries commerciales. Enfin, des

programmes de suivi de la qualité de l’environnement utilisent déjà la moule comme organisme

« sentinelle » au Canada dont Environnement Canada et le ministère des Pêches et Océans

Canada. Nos outils de diagnostic de la santé des moules seront donc particulièrement pertinents

pour y être intégrés.

Liens avec la présente demande :Plusieurs des paramètres proposés dans la présente demande,

dont le volet génomique, ont été mis au point grâce à cette stratégique.

Titre : Méthode alternative ou complémentaire au BeLPT

Organisme(s) : IRSST

Chercheur(s) / Michel Fournier, Claude Daniel, Jacques Bernier, Pauline Brousseau,

Montant / Durée (en mois) : 200,000 IRSST, 2010-2011

Problématique de santé et de sécurité du travail et objectifs : Le béryllium (Be) est connu

pour sa grande toxicité. Il peut provoquer suite à l’inhalation de poussières ou de vapeurs une

réaction de type allergique, la sensibilisation au béryllium (BeS) qui précèderait une

pneumopathie sévère dite maladie chronique du béryllium (MCB). En moyenne, 1 à 20% des

travailleurs exposés deviennent sensibilisés desquels 1 à 10 % développent la MCB. Il existe un

test de laboratoire couramment utilisé pour déceler la sensibilisation d’un travailleur au

béryllium; il s’agit du test de prolifération des lymphocytes en présence de béryllium le

"Beryllium Lymphocyte Proliferation Test" (BeLPT) Bien que ce test permette de déceler la

sensibilisation, il offre peu de valeur prédictive quant aux risques de développer une bérylliose.

En effet, un travailleur pourra avoir un BeLPT positif sans être nécessairement porteur d’une

bérylliose. Sachant qu’une bérylliose peut prendre jusqu’à 30 ans à se développer chez un

travailleur sensibilisé, il est évident que la comparaison BeLPT et bérylliose chronique n’est pas

acceptable (Kelleher et al., 2001). Le BeLPT est effectué couramment au Québec depuis 2002 et

certains résultats lors de suivi de travailleurs demeurent énigmatiques. Deux situations se sont

produites pour lesquelles nous n’avons pas encore d’explication. La première dans laquelle des

travailleurs porteurs de bérylliose mais dont les symptômes s’amélioraient avaient des tests de

BeLPT positifs, c’est-à-dire toujours la présence d’une sensibilisation confirmée or, au moment

du dernier test il s’est révélé négatif. La deuxième, dans laquelle des travailleurs porteurs de

bérylliose et dont les signes cliniques allaient en s’aggravant ont toujours eu des BeLPT négatifs.

Il devient donc primordial de développer un autre outil alternatif et/ou complémentaire au BeLPT

afin de déceler la sensibilisation au béryllium plus adéquatement, ce qui permettra d’une part de

rassurer les travailleurs quant à la détection plus efficace de la sensibilisation aider les

gestionnaires du dossier béryllium à mieux cerner l’ampleur du problème et d’autre part de faire

une étude scientifiquement plus acceptable de la valeur prédictive du BeLPT. Méthode : À partir

d’une cohorte de travailleurs pour laquelle nous avons comme objectif à priori d’obtenir 500

échantillons, chacun de ces échantillons sera utilisé dans cinq tests soit le BeLPT classique avec

incorporation de thymidine tritiée, l’immunoLPT en utilisant la cytométrie de flux, l’ÉLISPOT

avec lequel nous allons évaluer le nombre de cellules sécrétant spécifiquement de l’interféron-γ et

de l’interleukine-2 et un bilan sanguin par phénotypage et génotypage. Dans cette cohorte, nous

allons inclure des travailleurs non exposés au béryllium, exposés et non sensibilisés, exposés et

sensibilisés de même que des porteurs de bérylliose.

Liens avec la présente demande : Aucun

Titre : Centre Interinstitutionnel de recherche en écotoxicologie Organisme(s) : FQRNT Réseau stratégieue

Chercheur(s) / Michel Fournier, et 44 autres chercheurs.

Montant / Durée : $5,075,000, 2011-2017

Résumé :

La multiplicité des facteurs physiologiques et environnementaux susceptibles de moduler d'une

part la nature des agresseurs toxiques et d'autre part la réponse biologique à ceux-ci, rend

complexe la mise en évidence des effets multiples et insidieux directement reliés à leur présence

dans le système Saint-Laurent. Ces relations de cause à effet sont particulièrement difficiles à

identifier car elles sont reliées à des expositions chroniques et à des mélanges de contaminants.

De plus, il faut considérer qu'un certain nombre de manifestations néfastes peuvent être attribuées

à des actions indirectes de polluants sur les écosystèmes. Le CIRÉ s'est donné comme mission de

développer et de coordonner des programmes de recherche d'envergure, afin d'aborder cette

problématique avec une approche pluridisciplinaire. Les retombées de ces travaux, tout en

permettant de dégager une vision globale de la situation, fourniront les bases scientifiques à la

mise en place de solutions.

Cette mission englobe l'identification et la caractérisation de contaminants, le développement et

la validation de procédés et de stratégies d'intervention et la mise en place de programmes de

suivi. Plus spécifiquement, ces objectifs sont atteints par des activités relevant de plusieurs

domaines de recherche et d'ingénierie tels que l'identification et la caractérisation des

contaminants, l'étude de leur biodisponibilité, la modélisation de leur dispersion, la mise au point

et la validation de biomarqueurs cellulaires et moléculaires, le développement de procédés de

génie dans un contexte d'innocuité environnementale, etc. Ces activités sont réalisées de concert

avec des intervenants locaux, les divers paliers de gouvernements, ainsi que l'industrie. De plus,

le CIRÉ a mis sur pied de nombreux programmes de recherche à l'échelle internationale, afin

d'intéresser des chercheurs étrangers à nos problématiques de recherche et de leur donner accès à

de l'infrastructure particulière. Ces collaborations nous permettent aussi d'exporter notre savoir

faire et notre expertise.

Le CIRÉ, de par la structure de son fonctionnement et à travers l'interdisciplinarité, permet de lier

les producteurs et utilisateurs de données scientifiques et offre un cadre unique pour la formation

de personnel hautement qualifié. Le CIRÉ considère aussi que sa mission intègre la transmission

d'informations scientifiques vers le grand public.

La présente demande vise à assurer le financement d'infrastructure de recherche du Centre

interinstitutionnel de recherche en écotoxicologie ou CIRÉ. Le CIRÉ développe une

programmation de recherche qui cherche à comprendre les interactions entre les sources de

pollution, le transport et le devenir des polluants dans l'environnement ainsi que leurs impacts sur

les organismes vivants. Le CIRÉ travaille aussi au développement de procédés de traitement pour

prévenir et corriger les impacts négatifs des polluants. Ce centre regroupe les forces vives en

écotoxicologie et en toxicologie environnementale du Québec et de l'étranger. Il met à

contribution des équipes multidisciplinaires en provenance d'universités, de ministères, de

municipalités et de l'industrie permettant d'aborder les grands enjeux liés à la présence de

polluants dans l'environnement.

Liens avec la présente demande : Infrastructures de recherche

Titre : La bioindication adaptée à l'évaluation intégrée des cours d'eau en milieu agricole

Organisme(s) : CRSNG stratégique

Chercheur(s) / Catherine Jumarie et al.,

Montant / Durée : $747,000, 2011-20174

Résumé :

L'eau est une ressource naturelle essentielle à la vie dont la qualité et l'abondance sont affectées

par les activités anthropiques. Au Canada, 66% de la consommation d'eau est destinée aux

activités agricoles. Neuf des dix rivières les plus polluées du pays sont au Québec. Entre mai et

août, la rivière Yamaska rejette chaque jour dans le fleuve Saint-Laurent plus de 20 kg de

l'herbicide atrazine. Bien que les concentrations de pesticides mesurées dans l'eau soient

généralement en deçà des critères établis pour la protection de la vie aquatique, nous avons

observé que les grenouilles souffrent de graves problèmes de santé. C'est le cas d'invertébrés et de

poissons, ailleurs au Canada. Respecter ces critères ne permet donc pas de s'assurer de la qualité

de l'eau ni de souscrire au développement durable. Des éléments biotiques sont maintenant

intégrés aux programmes de surveillance, mais les bilans de santé des cours d'eau demeurent

incomplets. Notre objectif est de développer un ensemble d'outils permettant d'évaluer avec

justesse la qualité des cours d'eau en milieu agricole. Notre approche tiendra compte de la

contamination de l'eau, des diatomées, des invertébrés benthiques et des algues ainsi que de

l'expression de biomarqueurs mesurés chez des espèces sentinelles (grenouilles, moules,

poissons).Nos données générerons des outils de diagnostic tenant compte de la réelle pression

environnementale. Nous avons réuni des chercheurs universitaires, des experts provenant des

ministères impliqués dans la surveillance de la qualité de l'eau au Québec et au Canada, ainsi que

des organismes publics de bassins versants L'utilisation d'un éventail de biomarqueurs

judicieusement choisis et validés permettra aux scientifiques et décideurs en matière de

règlementation de mieux assurer la qualité de l'eau et la santé des écosystèmes aquatiques. Cela

se concrétisera par l'identification d'espèces plus sensibles et de biomarqueurs plus adaptés à la

pollution agricole diffuse et par le développement d'un modèle hydrologique prédictif. Nous

ferons aussi un bilan complet de l'état de santé actuel du bassin versant de la rivière Yamaska qui

dessert une importante population en eau potable et espérons expliquer le déclin de certaines

populations de poissons et d'amphibiens. .

Liens avec la présente demande : Aucun lien

RELATIONSHIP TO OTHER RESEARCH SUPPORT page

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I. CURRENT RESEARCH FUNDING 1. The human blood-epididymal barrier, an essential component of male fertility (CIHR Operating Grant; 2010-2015). D. Cyr (PI), and P. Chan. The objectives of this project are to assess the role of epididymal tight junctions in human male infertility. The project focuses on the development of human epididymal cell lines and assessing the pattern of expression of claudins in the epididymis. While there is no financial overlap between this medically oriented project on humans. There is no financial overlap with the present study. PROPOSAL ABSTRACT Approximately 12-15% of couples experience fertility problems and male infertility accounts for half of these cases. In patients with normal spermogram and diagnosed as idiopathic infertile, it is thought that post-testicular factors are responsible for the infertility, as the problem appears associated with the process of sperm maturation. While spermatozoa are formed in the testis via spermatogenesis, these spermatozoa do not have the ability to fertilize or swim. These functions are acquired in the epididymis. If we are to understand the normal and pathological conditions attributable to human epididymal function, we must establish the physiological, cellular and molecular regulation of the human epididymis, develop tools to characterize these functions and develop clinical strategies to improve treatment. This study will address two critical function of human epididymal physiology: the molecular regulation of the tight junctions which make up the blood-epididymis barrier, and the regulation of epididymal sperm maturation. Our hypothesis is that cellular and molecular regulation of proteins, including those which comprise epididymal tight junctions and essential secreted proteins, represent clinical targets that can be exploited to improve epididymal function and fertility in obstructive and non-obstructive azoospermic infertile patients following epididymostomy. To address this hypothesis we intend to understand how Cldns, the protein which make up tight junctions, are regulated in the human epididymis and using recently developed cell lines, understand epididymal sperm maturation. STUDENTS IMPLICATED IN PROJECT: Sebastien LeQuerrec, Berthes Youness (starting May 2011), Marion Mandon (starting May 2011) 2. FRSQ-RRSE Cell and Molecular Effects Unit (2009-2013). D. Cyr. The objective of this FRSQ Environmental Health Network (RRSE) service platform is to provide genomic, cell, and molecular support to members of this FRSQ Network. The unit offers equipment and expertise. The funding is part of a larger network funding which encompasses 4 different platforms. While there is no financial or intellectual overlap between the unit and the proposed research, much of the platform infrastructure will be put to contribution for this study. PROPOSAL ABSTRACT (1 page Summary)

L’objectif stratégique du RRSE consiste à approfondir les connaissances de l’impact de l’environnement sur la santé en intégrant des outils et des approches spécialisés pour aborder globalement des problématiques de santé environnementale. L’exposition constitue une étape essentielle qui détermine en bonne partie non seulement l’apparition, mais également la nature et l’intensité des effets néfastes à la santé qui sont associées aux contaminants chimiques. La structure primaire du RRSE a été développée autour de quatre regroupements stratégiques complémentaires, dont trois sont associés au thème de l’exposition aux contaminants, soit les biomarqueurs, la mesure et la modélisation de l’exposition et l’épidémiologie environnementale. Le quatrième regroupement, la toxicologie cellulaire et moléculaire, est axé vers la mesure de manifestations précoces de la toxicité qui est applicable à divers tissus et qui ouvre la voie à l’étude de diverses pathologies. Le RRSE propose de maintenir et de développer ses quatre regroupements et les infrastructures qui


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leurs sont associées, lesquelles apparaissent plus que jamais essentielles pour réaliser en synergie des activités de recherche convergentes dans le cadre des grands projets/programmes communs.

En s’appuyant sur son plan de développement, le RRSE propose d’appliquer les outils développés au cours des quatre dernières années à l’étude des liens entre l’exposition aux contaminants environnementaux et divers désordres et pathologies, dans le cadre de grandes études populationnelles qui sont déjà en cours ou qui démarreront sous peu telles CARTaGÈNE, l’IRNPQEO, MIREC-ID et l’Enquête canadienne sur les mesures de santé. Ces études comportent toutes des biobanques qui comprennent différents échantillons biologiques, lesquels serviront à caractériser l’exposition, de concert avec l’évaluation par questionnaire et la modélisation toxicocinétique. Les chercheurs du RRSE sont parmi les leaders mondiaux dans l’utilisation combinée de ces différentes approches pour permettre l’évaluation de l’exposition pendant différentes périodes cruciales de la vie en lien avec les effets toxiques étudiés. Les articles récents publiés par Verner et collègues dans la revue phare Environmental Health Perspectives en sont des exemples éloquents. Cette caractérisation poussée de l’exposition, jumelée à la mesure de biomarqueurs d’effets précoces dont l’expression génique dans des cellules sentinelles, permettront des avancées importantes en épidémiologie environnementale.

Notre stratégie s’appuie avant tout sur les acquis des quatre dernières années et permettra de mettre à contribution les expertises de nos quatre plateformes auxquelles viendront s’ajouter de nouveaux outils. Les grands projets seront une occasion unique d’intégrer les savoirs et compétences des membres du RRSE. La structure de nos activités permettra aussi d’assurer la formation des étudiants, dans une perspective multidisciplinaire, par le biais d’un programme de formation inter-plateformes qui leur permettront de se familiariser avec des approches différentes de celles utilisées à l’intérieur de leur propre projet. Même si les activités de recherche, soutenues par nos infrastructures, sont au centre de notre mission et visent à développer de nouvelles connaissances, nous sommes néanmoins conscients du rôle primordial que se doit de jouer le RRSE au niveau de la diffusion et du partage des connaissances. C’est pourquoi nous allons continuer à promouvoir le recours à notre site Internet et proposer à nos membres chercheurs et à nos étudiants des ateliers de formation. Certains ateliers pourront même être offerts à divers intervenants du milieu, notamment les gestionnaires et scientifiques des gouvernements provincial et fédéral. En résumé, le RRSE entend au cours des quatre prochaines années faire les efforts nécessaires pour promouvoir et assurer le développement de la recherche en santé environnementale au Québec et la diffusion des connaissances nouvelles, notamment par l’organisation de conférences de prestige et la préparation d’articles de vulgarisation. STUDENTS IMPLICATED IN PROJECT: None 3. Assessing the Impacts on aquatic organisms exposed to emerging contaminants in wastewater discharge. (2010-13; Canadian Water Network) C. Metcalfe (PI) and others. The objective of this project is to study the effects of municipal wastewater effluent on fish and molluscs in the St. Lawrence, Grand and Saskatchewan Rivers. Studies will define mixing zones and provide data on the effects of endocrine disrupting chemicals on DNA integrity, gene expression, chemical levels in these rivers and determine how aquatic organisms are impacted. There is no overlap with the present study. PROPOSAL ABSTRACT We have developed a proposal for the practical monitoring of emerging contaminants originating from sewage treatment plants with different waste management strategies and their effects on fish species in large rivers. Our main question is whether or not there is a direct link between


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effluent exposure and biological effects in free-living fish. This national research team is supported internationally and will develop an approach that specifically addresses the EC-1 topic defined by the Canadian Municipal Water Management Research Consortium (CMWMRC) and the Canadian Water Network (CWN). Collections and surveys of fish body weights and gonad sizes will be completed by Sept. 2010. By measuring the concentrations of natural and synthetic therapeutic estrogens and fluoxetine (the active ingredient in Prozac) we provide added value to existing pharmaceutical monitoring because we will determine if these contaminants bioconcentrate in sport fish (by May 2012). There is cause for concern and we have focused this proposal on these 2 pharmaceutical classes because they are model compounds and are representative contaminants of different solubilities. We already have strong data showing they impact fish reproduction so there is a possible ecosystem health risk. Our approach will provide an estimate of the time frame (weeks, months, one-year) over which individual fish will have been exposed to sewage effluents and EDCs. Isotope analyses common in ecological food-web studies are applied here for the first time to the problem of emerging contaminants to better estimate the extent over which fish populations are affected (completed by Dec. 2011). Biomarkers that assess the negative effects of pharmaceutical on fish reproduction will be validated (by May 2012). Isotope analysis involves inexpensive and simple tissue preparation (freezing and drying will suffice). This approach in combination with existing contaminant monitoring programs could therefore become a simple but powerful tool for monitoring ecosystem health. We will report our findings at workshops (Jan. 2011, June 2012) and prepare scientific articles for publication (by Sept. 2012). Streamlining of the contaminant and environmental monitoring programs will be important to municipal water managers and provincial regulators because they must balance the financial cost of new approaches with their long-term strategies to control operational costs. ` STUDENTS IMPLICATED IN PROJECT: Jennifer Airstakaitis 4. Cellular communication in the epididymis, an essential component of sperm maturation (2002-10; currently in renewal; NSERC Discovery Grant) D. Cyr (PI). A crucial aspect of sperm maturation is the fact that the epididymal epithelium must coordinate the different regions of epididymis in order to modify the luminal environment necessary for sperm maturation. The overall objectives of this research program are to understand the role and regulation of gap junctional communication in the epididymis and the role of this cellular communication in sperm maturation. There is no financial overlap with the present study. PROPOSAL ABSTRACT Gap junctions are essential structural components for direct communication between neighboring cells. Communication between adjacent cells is critical for coordinating cellular function within tissues. Spermatozoa are formed in the seminiferous tubules of the testis. In mammals, testicular spermatozoa cannot swim or fertilize. These functions are acquired in the epididymis during sperm maturation. A crucial aspect of sperm maturation is the fact that the epididymal epithelium must coordinate the different regions of epididymis in order to modify the luminal environment in which the sperm bathe and mature. The overall objectives of this research program are to understand the role and regulation of gap junctional communication in the epididymis and the role of cellular communication in sperm maturation. The current short-term objectives are to understand the regulation of the proteins responsible for intercellular communication, the connexins (Cxs) and pannexins (Panxs), in the epididymis. We first intend to identify the signalling pathways regulating Cxs during the differentiation of the epididymis, and determine if Cxs are critical to this process. We will then assess how Panxs are regulated in the epididymis and determine if RNA processing plays an essential role in this regulation. Finally, we will identify the factors that regulate the expression


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of epididymal Cx and Panx genes, and show how these factors are themselves regulated. By addressing these crucial fundamental aspects of cellular communication and regulation of gene expression in the epididymis we expect to develop to better understanding of the regulation of cellular communication, epididymal function and sperm maturation.

STUDENTS IMPLICATED IN PROJECT: Steven Jones (tech); two students to be recruited II. FUNDING CURRENTLY APPLIED FOR 5. Role of the ubiquitin system in sperm development and function. (2011-16; CIHR Operating Grant) S. Wing (PI). The ubiquitin-proteosome system has been shown to be important for regulating the pattern of protein expression during spermatogenesis. However the role of ubiquinating enzymes in spermatogenesis is poorly understood. The objective of this study is to characterize the role of ubiquinating enzyme, USP2, in spermatogenesis. Using USP2 knockout mice we intend to demonstrate the role of ubiqitination on spermatogenesis and male fertility. There is no overlap with the present application. PROPOSAL ABSTRACT Infertility is a common problem that is increasing, particularly in Western societies. Approximately 20-25% of infertility cases are due to the male partner and ~30% of additional cases are due to abnormalities in both partners. The causes of male infertility remain largely unknown. In order to fertilize eggs, sperm must undergo normal development and maturation. This involves the conversion of undifferentiated stem cells in the testis into mature elongated spermatids. These spermatids undergo further maturation in the epididymis and activation upon ejaculation into the female reproductive tract. Here the activated sperm acquire the necessary motility to move through the reproductive tract and then penetrate the outer layers of the egg structure to fuse with the egg. In order for proper development and maturation of spermatids to take place, proteins that are not required in the final sperm cells must be destroyed. My laboratory is studying the mechanisms by which the nonrequired proteins are degraded in developing spermatids. Defects in this breakdown of proteins may lead to abnormal development of sperm and infertility. Indeed we have recently observed that mice lacking an enzyme in this pathway of protein breakdown are almost completely infertile and their sperm have defective motility. Defects in sperm motility are found in the majority of infertile men and so this research may lead to new approaches to treating infertility or new contraceptive methods in men. STUDENTS IMPLICATED IN PROJECT: One to be recruited 6. Generation of new genetic tools for studying epididymal sperm maturation (2011-14. FQRNT Team Grant) D. Cyr (PÎ), R, Viger, N. Pilon. The objective of this study is to develop Cre-LoxP transgenic mice that specifically target different cell types of the epididymis. These transgenic animals could eventually be used to selectively knockout the expression of a gene in specific cell type and region of the murine epididymis. This study has no overlap with the present application. PROPOSAL ABSTRACT Les spermatozoïdes testiculaires ne peuvent nager ni féconder. Ces fonctions sont acquises dans l’épididyme. Même si nous connaissons l’importance de l’épididyme dans la maturation des spermatozoïdes, les aspects moléculaires de l’épididyme demeurent peu connus. L’absence de modèle animale pouvant servir à étudier les gènes de l’épididyme limite l’avancement des


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connaissances sur la maturation des spermatozoïdes. Afin d’adresser cette limitation, notre objectif est de développer des nouveaux outils génétique pour l’analyse de la maturation des spermatozoïdes dans l’épididyme. Nous proposons de : 1. Développer des lignées de souris transgénique pouvant cibler des gènes spécifiquement exprimé dans différentes régions de l’épididyme; 2. Identifier des gènes dans les cellules basales dont le promoteur servira à générer des souris transgénique. Notre stratégie vise à utiliser les promoteurs de deux gènes, la Lcn-5 (lipocaline-5) et la Lcn-8 (lipocaline-8), pour générer des souris transgéniques exprimant la Cre-recombinase sous le contrôle de ces promoteurs. La Lcn-5 est exprimée dans les cellules principales de la tête distale de l’épididyme tandis que la Lcn-8 est exprimée dans le segment initial. Pour cibler le transgène dans la région proximale de la tête de l’épididyme, nous utiliserons la région régulatrice du gène Cst8 (cystatin-related epididymal spermatogenic protein) dont nous remplacerons le 1er exon du gène par un ADNc Cre, afin que Cre soit régulé par le locus de gène Cst8. Finalement nous allons caractériser l’expression de gènes des cellules basales afin de développer une stratégie pour générer des souris transgéniques ciblant ces cellules. STUDENTS IMPLICATED IN PROJECT: Two to be recruited

SIAH, Ahmed

Title: Immuno-competency in Mytilus edulis from the larvae to the adult stage and its

sensitivity to bacterial challenge

Funding source: NSERC Strategic

Investigators: Michel Fournier (PI), Gauthier-Clerc Sophie, Pellerin Jocelyne,

Lemarchand Karine, Ahmed Siah, Auffret Michel, Danger Jean-Michel, Moraga Dario.

Budget/Duration: $470,000, 2009-2011

Abstract:

The main goal of this project is to identify the immune effectors involved in the control

of the immune system in the bivalve mollusk, Mytilus edulis. Specifically, this work aims

to identify the cellular and humoral immune defence in Mytilus edulis and its sensitivity

to a bacterial challenge. The expression of genes will be quantified in hemocytes from

healthy mussels and compared to challenged blue mussels to clearly detect the induction

of genes involved in immune defence.

Relation with current proposal: In this project, a cDNA library from blue mussel hemocytes was generated and

sequenced. More than 20,000 unique ESTs were annotated and will serve as a reference

transcriptome for Mytilus edulis' hemocytes. This project will provide the grassroots for

the genomic analyses suggested in the proposed project.

Title: The development of techniques and mitigation strategies for the management of

invasive tunicate species fouling aquaculture farms

Funding source: Atlantic Innovation Fund

Investigators: Russ Kerr (PI), Ahmed Siah, Jeffrey Davidson

Budget/Duration: $3,773,185/2008-2011

Abstract:

Monitoring is essential to the management of aquatic invasive species both in terms of

prevention and treatment. Our group of research is focusing on the development of an

effective tunicate diagnosis method for commercialization. In this project, we aim at

developing a diagnostic kit through microarray analysis, innovative molecular bead-based

technology and flow cytometry development. This new technology will enable a high

sensitivity and high specificity of detection, which is essential for a more accurate and

rapid monitoring of the presence of tunicate species in aquatic areas. As an early warning

detection, the kit will be field-tested and will assist the mussel farm industries to prevent

and treat tunicates more efficiently. The prevention and treatment are the goals for the

other two modules of the project.

Relation with current proposal: No direct link with the suggested proposal

Title: Innovative diagnostic tool for mussel health management

Funding source: PEI Provincial Innovation Fund

Investigators: Ahmed Siah (PI)

Budget/Duration: $32,000, June 2010-June 2011

Abstract: This project is aiming at developing a diagnostic kit, which would be the first

early-stage diagnostic tool for mussel health. The outcome of this project will be a set of

SIAH, Ahmed

genes coated on beads to be used on a flow cytometry platform. The new advances on

bead-based flow cytometry technology enable a multiplex analysis and then a screening

of a specific target at a large scale of the genome. This technology offers the advantage of

screening the messengers with a rapid, high sensitivity/specificity and high throughput

analysis. The targeted users fot this assay are government agencies such as the

Department of Fisheries and Oceans, Canadian Food Inspection Agency, the aquaculture

industry both in Atlantic Canada and abroad, the aquaculture associations, as well as

mollusk growers.

Relation with current proposal: No direct link with the proposed project

Title: Development of high-throughput diagnostic assay for simultaneous detection of

salmonid viruses using fluidic bead-based molecular assay technology.

Funding source: PEI Provincial Development and Discovery Fund

Investigators: Ahmed Siah (PI), Carmencita Yason (PI), David Groman

Budget/Duration: $450,000/2010-2012

Abstract: This project proposes to develop a high-throughput (up to 96 diagnostic

samples) multiplex assay for simultaneous detection of salmonid viruses (up to 100

molecular targets). The goal is to produce a molecular tool that will be able to

simultaneously detect economically important aquatic viruses for disease diagnosis, bio-

surveillance, and routine infectious agent monitoring with an acceptable turnaround time.

This assay will be validated as a diagnostic tool and its performance will be compared

with existing diagnostic tests such as PCR and virus isolation. The new product will be

offered as a test at AVC Diagnostic Services.

Relation with current proposal: No link with the proposed project

ST-PIERRE, Yves

Title : The role of galectin-7 in mammay tumorigenesis and metastasis.

Agency : CIHR

PI(s) / Yves St-Pierre, INRS-Institut Armand-Frappier

Amount/Period : $292, 899, 2008-2011

Abstract : This project aims to further our knowledge of the molecular determinants of invasion

and metastasis, which are critical steps in the progression of cancer. We will examine how

galectin-7 is implicated in metastasis of breast cancer and study its expression in human breast

tumor material. A particular attention is paid to p53 and the role of its mutant form in controlling

galectin-7 expression and other cancer genes. This information will increase our understanding of

the development of cancer and may yield better markers for the pathological assessment of

cancer.

Overlap with the current project: These studies help to better understand the frequency and the

nature of specific mutations in the p53 gene that have relevance for specific types of cancer.

Title : Functional relationship between egr-1 and MMP-9 during lymphoma progression.

Agency : Cancer Research Society of Canada

PI(s) / Yves St-Pierre, INRS-Institut Armand-Frappier.

Amount/Period : $119, 600, 2010-2012

Abstract : The general goal of our project is to better understand the molecular mechanisms used

by lymphoma cells to interact with stromal cells during during lymphoma progression, including

the growth of thymic lymphoma and dissemination of lymphoid tumors at distant sites. For this

purpose, we are using a well-characterized pre-clinical model of T-cell lymphoma combined with

genetically-engineered transgenic mouse models. We are specifically focusing on molecular

mechanisms that suppress the expression of metastatic genes that have been associated with

aggressive forms of human lymphomas, such as non-Hodgkin’s lymphoma (NHL). Some of

these genes, including mmp genes, have also been involved at different steps of metastasis in

other types of cancer. We thus believe that our project has high not only high relevance in the

field of lymphoid tumors, but it may also have relevance in the field of tumor progression in

general.

Overlap with the current project: None.

SPEAR, Philip

1. Titre : A complete solution for biological sample clean-up and solvent evaporation in

environmental contaminant analysis (J. Verreault demandeur) 2. Montant (annuel et durée de subvention)

Année : 2011 65 112 $ (demandé)

3. Objectifs et brève description de la recherche :

Demande d’équipement nécessaire pour la préparation d’échantillons biologiques

pour l’analyse des concentrations en contaminants organohalogènes. Un système

chromatographie en perméation de gel avec auto-échantillonneur et un appareil

d’évaporation rotative des solvants. 4. Nombre et statut d’étudiants impliqués

0 5. Lien (par ex. résultats obtenus servant de prémisse à la présente demande) ou absence de lien (pas de duplication entre les projets) avec la présente demande stratégique :

Aucun lien avec la présente demande.

______________________________________ 1. Titre : Renouvellement du Centre TOXEN : Infrastructure du centre de recherche

universitaire 2. Montant (annuel et durée de subvention) Année : 2011-2012 70 400 $ (demandé) Année : 2012-2013 70 400 $ (demandé) Année : 2013-2014 70 400 $ (demandé) 3. Objectifs et brève description de la recherche :

Subvention pour le fonctionnement du Centre de recherche TOXEN qui sert à

maintenir une technicienne, secrétariat et support pour les projets de recherche associés

au Centre. 4. Nombre et statut d’étudiants impliqués :

Plusieurs étudiants bénéficient indirectement de cette infrastructure. 5. Lien (par ex. résultats obtenus servant de prémisse à la présente demande) ou absence de lien (pas de duplication entre les projets) avec la présente demande stratégique

Cette subvention n’est pas directement reliée à la demande présente, mais est un

support général pour les activités du groupe. __________________________________ 1. Titre : Identifier les impacts toxiques et écosystèmiques des floraisons algales.

Centre interinstitutionnel de recherche en écotoxicologie (CIRÉ) – Responsable : Pauline Brousseau, Centre TOXEN et INRS-IAF

SPEAR, Philip

2. Montant (annuel et durée de subvention) Année : 2010-2011 58 000 $ (octroyé) Année : 2011-2012 60 000 $ (octroyé) Année : 2012-2013 60 000 $ (octroyé) 3. Objectifs et brève description de la recherche :

Recherche en collaboration avec Environnement Canada au sujet des floraisons

algales. La recherche comprend plusieurs volets dont le développement des méthodes

pour l’analyse des substances toxiques produits par les cyanobactéries, investigations sur

les conditions environnementaux affectant la synthèse des toxines, les modèles

toxicocinétiques de l’accumulation des toxines chez les poissons. 4. Nombre et statut d’étudiants impliqués :

une chercheure postdoctorale 5. Lien (par ex. résultats obtenus servant de prémisse à la présente demande) ou absence de lien (pas de duplication entre les projets) avec la présente demande stratégique.

Il n’y a pas de lien direct, cependant certains sites d’échantillonnage sur le

système Yamaska sont communs entre les deux projets. Ceci pourra économiser du temps

et les coûts sur le terrain. _________________________________

1. Titre : Centre interinstitutionnel de recherche en

écotoxicologie

Michel Fournier et 30 autres

FQRNT : Regroupements stratégiques 2. Montant (annuel et durée de subvention)

2011 845,000 (demandé)

2012 845,000 (demandé)

2013 845,000 (demandé)

2014 845,000 (demandé)

2015 845,000 (demandé)

3. Objectifs et brève description de la recherche : Cette subvention porte sur la création d’un centre de recherche FQRNT, le Centre

interinstitutionnel de recherche en écotoxicologie (CIRÉ). Parmi une cinquantaine de

projets identifiés dans cette demande, trois impliquent la pollution agricole. 4. Nombre et statut d’étudiants impliqués

La demande FQRNT ne comprend pas du support pour les étudiants, mais trois étudiants

à la maîtrise effectueront des projets de recherche associés à ces projets.

SPEAR, Philip

5. Lien (par ex. résultats obtenus servant de prémisse à la présente demande) ou absence de lien (pas de duplication entre les projets) avec la présente demande stratégique

La demande FQRNT est un compliment à la présente demande CRSNG Stratégique. Plus

spécifiquement, le CIRÉ supporterait des projets concernant l’évaluation des impacts

chez les poissons, les amphibiens et le bivalve, Elliptio sp. Le support FQRNT aiderait

donc à défrayer certains coûts associés aux salaires de techniciens, des déplacements sur

le terrain ainsi qu’à la collecte et à l’analyse des échantillons.

_________________________________

1. Titre : La bioindication adaptée à l'évaluation intégrée des cours d'eau en milieu

agricole

Catherine Jumarie et 11 autres CRSNG Stratégique 2. Montant (annuel et durée de subvention)

2011 180,000$

2012 180,000$

2013 180,000$ 3. Objectifs et brève description de la recherche

Ce projet vise une nouvelle stratégie qui intègrerait des réponses biologiques à plusieurs

niveaux trophiques dans l’évaluation des influences de l’agriculture. Cette stratégie serait

donc un support aux «normes» ou «critères» basés sur les concentrations dans l’eau de

substances individuelles (ex. : pesticides, nutriments). 4. Nombre et statut d’étudiants impliqués

Sont demandés : deux étudiants à la maîtrise et un technicien à temps partiel. 5. Lien (par ex. résultats obtenus servant de prémisse à la présente demande) ou absence de lien (pas de duplication entre les projets) avec la présente demande stratégique Il n’y a as de lien direct entre cette demande CRSNG Stratégique et la présente demande.

Les objectifs, les espèces d’animaux et les sites d’échantillonnage sont différents. _________________________________

Title: Placental melatonin system: functions and mechanisms of action

Organism(s): NSERC – discovery grant

PI: C Vaillancourt

Amount: $150 000 (2009-2014)

Abstract: The human placenta acts as a central regulator of maternal and fetal physiology. Although

the role of melatonin on fetal development has been the subject of a number of studies, data on its

role in placental physiology are scarce. We have demonstrated that human placenta expresses

melatonin receptors. We have also shown the presence and activity of the enzymes responsible for

melatonin synthesis and placental melatonin production in placental trophoblast cells. These results

strongly suggest an important paracrine, autocrine and/or intracrine role for melatonin and its

receptors in placental function and development and, consequently, in pregnancy and fetal health.

However, the role of melatonin in placental function and development is still obscure. Thus, our

research program aims to elucidate the role and cellular mechanism of melatonin and its receptors in

placental endocrine function as well as on trophoblast development. That is, to better understand the

role of this trypthophan-derived hormone and its receptors in placental health, and by consequence in

pregnancy well-being and fetal development and to develop a new human cellular model to study the

regulation and function of melatonin and its receptors on normal and abnormal cellular processes and

open the way for new preventive and therapeutic approaches in these areas for variety of disorders

including obstetric complications, depression, cancer, and neurodegenerative diseases. Overlaps with the present proposal: None

Title: The Iowa Flood Study: Perinatal and Infant Effects of a Natural Disaster

Organism(s): National Institute of Mental Health - RAPID grant post-disaster program

PI: M O'Hara (UIowa)

Co-PIs: C Vaillancourt + 7 others

Amount: $ 300 000 (2009-2011)

Abstract: Stressing pregnant female animals result in damage to the foetus, subsequently

affecting later development. Studying prenatal maternal stress (PNMS) in humans is hampered

by ethical constraints: researchers cannot randomly assign pregnant women to stress and no-

stress conditions. Disasters create natural experiments in which the degree of stress may be

randomly assigned to large numbers of pregnant women. In January 1998, Southern

Quebecexperienced the worst disaster in Canadian history, an ice storm which knocked out

electrical power to more than 3 million people for as long as 45 days. McGill researchers

recruited over 150 pregnant women in June 1998 and have been following the women and the

development of their children ever since. Results show that more severe in utero exposure to

PNMS is associated with worse cognitive, behavioural, motor, and physical development during

infancy and earlier childhood. Yet Project Ice Storm has weaknesses: there were no pre-disaster

measures on the women, the sample size has diminished to about 140 families 10 years later, and

no biological samples (i.e., fetal and maternal blood, placenta, umbilical cord), needed to test

hypotheses about the way PNMS effects the foetus, were obtained. In June 2008, Iowa

experienced the worst flooding in 50 years. Before the flooding, University of Iowa researchers

had already recruited 135 pregnant women and assessed their psychological functioning, coping,

and social support to study the effects of everyday stress on pregnancies. The goal of the current

grant is to increase our understanding of the effects of PNMS on pregnancy and fetal outcomes,

and on early childhood development. An additional 300 women will be recruited to participate.

The current grant will allow us to test biological markers to determine the manner in which

PNMS influences development. It will also allow for a thorough prospective follow-up of

mothers and children annually between the ages of 1 and 5 years

Overlaps with the present proposal: None

Title: The Iowa Flood Project: Effects of prenatal maternal stress on pregnancy outcomes and

infant development Organism(s): CIHR – operating grant PI: S King

Co-PIs: C Vaillancourt + 6 others

Amounts: $1 800 000 (2009-2013)

Abstract: Stressing pregnant female animals result in damage to the foetus, subsequently

affecting later development. Studying prenatal maternal stress (PNMS) in humans is hampered

by ethical constraints: researchers cannot randomly assign pregnant women to stress and no-

stress conditions. Disasters create natural experiments in which the degree of stress may be

randomly assigned to large numbers of pregnant women. In January 1998, Southern

Quebecexperienced the worst disaster in Canadian history, an ice storm which knocked out

electrical power to more than 3 million people for as long as 45 days. McGill researchers

recruited over 150 pregnant women in June 1998 and have been following the women and the

development of their children ever since. Results show that more severe in utero exposure to

PNMS is associated with worse cognitive, behavioural, motor, and physical development during

infancy and earlier childhood. Yet Project Ice Storm has weaknesses: there were no pre-disaster

measures on the women, the sample size has diminished to about 140 families 10 years later, and

no biological samples (i.e., fetal and maternal blood, placenta, umbilical cord), needed to test

hypotheses about the way PNMS effects the foetus, were obtained. In June 2008, Iowa

experienced the worst flooding in 50 years. Before the flooding, University of Iowa researchers

had already recruited 135 pregnant women and assessed their psychological functioning, coping,

and social support to study the effects of everyday stress on pregnancies. The goal of the current

grant is to increase our understanding of the effects of PNMS on pregnancy and fetal outcomes,

and on early childhood development. An additional 300 women will be recruited to participate.

The current grant will allow us to test biological markers to determine the manner in which

PNMS influences development. It will also allow for a thorough prospective follow-up of

mothers and children annually between the ages of 1 and 5 years.


Title: Integrating gender and sex in health and environment research :development of new

methodology

Organism(s): CIHR – Emerging team grant : gender, sex and Health

PI: D Mergler

Co-PIs: C Vaillancourt + 22 other

Amount: $1 000 000 (2009-2013)

Abstract: Although it is common knowledge that boys and girls, men and women are

biologically different and that their experiences bring them into contact differently with their

physical and social environments throughout their lifespan, research in environment and health

has been slow to translate this knowledge into methodologically sound studies that adequately

consider sex and gender. The paucity of information in this area has vital implications for health,

health services and preventive intervention strategies for children, adolescents, family members

involved in paid and non-paid work, and the elderly. The overall objective of this proposal is to

integrate gender/sex considerations into research in health and environment throughout the

lifespan. To achieve this goal, we propose to set up a pan-Canadian interdisciplinary research

tem, building on recognized Canadian expertise in environment and health and in quantitative

and qualitative methods to take account of gender and sex. The emerging team will (i) develop

and refine interdisciplinary quantitative and qualitative methods for integrating gender/sex

considerations in Canadian and international research in environment (including workplace and

health; (ii) establish strong collaborations with community-based and public health partners

working in environment and health, in order to come to a richer understanding of the real-life

mechanisms linking environmental health to sex and gender; (iii) train and mentor young

researchers; (iv)initiate multi-centre, multi-disciplinary research demonstrating the scientific

importance of integrating gender/sex considerations into research and intervention in health and

environment. We propose a participatory structure with partners working in the field of

environment and health and a 5-year progressive research and activities plan grounded in on-

going research in environment and health across Canada.


Title: Niveau de stress des femmes enceintes vivant en milieux linguistiques minoritaires et l'état

de santé du bébé

Organism(s): Ministère de l'éducation du QC - Coopération NB-QC

PI: C Vaillancourt

Coll.: J Normandeau (UdeMoncton)

Amount: $ 7 500 (2009-2011)

Abstract: L’ensemble des études qui ont examiné l’impact de la situation linguistique

minoritaire sur la santé des femmes enceintes et l’état de santé des bébés a surtout été mené aux

États-Unis, dans un contexte historique et socioculturel très différent des francophones du

Canada. Les femmes enceintes éprouvent le besoin de communiquer leurs émotions et leurs

sentiments durant les visites prénatales. Une étude révèle que les femmes qui ont indiqué

éprouver des difficultés de communication (ex. : due à la barrière de langue) avec les

professionnels de la santé en période prénatale ont un risque plus élevé de souffrir de dépression

postnatale. Ainsi, la langue revêt une dimension particulièrement grande lorsqu’il s’agit des

questions de santé où la relation patient-professionnel de la santé joue un rôle prépondérant. Les

communautés francophones en situation minoritaire représentent à peu près 3,18 % de la

population canadienne et 32 % des Néo-Brunswickois (Statistiques Canada, 2002). Alors que la

loi sur la santé prône l’accès universel à des soins de qualité, il n’est pas clair dans quelle mesure

les minorités de langue officielle peuvent recevoir des services de santé culturellement adaptés

dans leur langue. Une revue de la littérature faite pour Santé Canada suggère que les usagers du

système de soins qui ne peuvent pas communiquer dans leur langue n’ont pas le même accès ou

la même qualité de soins que leurs concitoyens. La barrière linguistique réduirait le recours aux

services préventifs, limiterait l’accès à tous les services basés sur la communication

(accompagnement de la grossesse, santé mentale, services sociaux…), et contribuerait à accroître

le recours aux services d’urgence et à des examens supplémentaires pour compenser les

problèmes de communication. Par exemple, l’étude coordonnée par la Fédération des

communautés francophones et acadiennes du Canada (2001) a révélé que les services de santé en

français dans divers lieux de prestations de services étaient de 3 à 7 fois moins accessibles aux

francophones dans 71 régions du pays où ils vivent en situation minoritaire. Dans ce contexte, ce

projet de coopération vise à recueillir de l'information sur les expériences de maternité des

femmes francophones du NB. Les résultats de cette étude seront utilisés afin d'aider à

l'amélioration des soins de santé et des informations à la disposition des femmes à cette époque

de leur vie.


Title: Comité de Programme TOXAIR: Lieu d’enfouissement technique (LET)-Lachenaie

Organism(s): FQRNT- CIRÉ

PI: C Vaillancourt

Amount: $20 235 (2009-2011)

Abstract: Le comité de programme regroupe scientifiques et gestionnaires et a pour mandat

: i) d’assurer la gestion et la réalisation des projets de recherche, en accord avec les besoins des

utilisateurs, ii) d’évaluer les projets de recherche des sous-comités.


Title: Système de PCR en temps réel en microplaque à haute

Organism(s): NSERC-OIR -1

PI: C Vaillancourt

Amount: $91 005 (2010-2011)

Abstract: La technique d'amplification en chaîne par polymérisation (PCR) en temps réel a

permis une avancée majeure de la recherche dans le domaine des sciences naturelles et

biomédicales. Cet appareil est indispensable pour effectuer des travaux de recherche compétitifs

dans le domaine de l'expression génique, de la signalisation cellulaire et des mécanismes

normaux ou pathologiques de l'activité cellulaire. Cette demande vise l'acquisition d'un système

de PCR en temps réel à haut débit, soit la technologie quantitative la plus performante sur le

marché pour mesurer le taux d'expression des gènes. De façon spécifique, cet appareil permettra

l'avancement de nos travaux de recherches dans les domaines de la reproduction, de la

toxicologie, de l'endocrinologie, du cancer, de la survie neuronale, de l'immunologie, de

l'inflammation et du stress oxydatif. Cette puissante technique permet de quantifier et de

comparer l'expression de différents gènes avec précision ou encore de quantifier leur nombre de

copies. En outre, ce système à haut débit nous permettra de prendre avantage des nombreux

nouveaux produits commerciaux (PCR Array) pour le criblage et l'étude approfondie des

modulations de l'expression génique. Le système de PCR en temps réel est indispensable pour la

réalisation de nos programmes de recherche, tous subventionnés par le CRNSG, qui exigent

l'évaluation des modulations génétiques des différents intervenants cellulaires et membranaires à

l'étude dans les différents systèmes biologiques : cellules trophoblastiques et placentas, follicules

ovariens, maladies infectieuses, neutrophiles, cellules cancéreuses, cellules T et dendritiques et

neurones. L'appareil demandé servira à de nombreux utilisateurs (au moins 8 équipes de

recherches, plus de 30 étudiants, post-doctorants, assistants de recherche et techniciens) et des

projets de recherche multidisciplinaires ; la rapidité, la sensibilité et la versatilité sont donc

primordiales. Cet appareil permettra d'obtenir des résultats plus rapidement et à nos étudiants

d'acquérir une formation à la fine pointe de la technologie. Le système de PCR en temps réel à

haut débit est donc de première importance pour l'ensemble de nos travaux et la formation de

personnel hautement qualifié.


APPENDIX A (Form 101)Environmental Impact )

to determine any potential environmental effects.The information contained in this form will be used by NSERC

(see the "Requirements for Certain Types of Research" in the NSERCPROTECTED WHEN COMPLETED


NOTE: There is a potential to generate several Appendices A.in the space provided in the upper right corner of the form. IF YOU FORESEE THE NEED FOR MORE THAN 3 (THREE)

(613) 995-8079, OR BY E-MAIL AT [email protected].

SEND ONEORIGINAL ONLY

DO NOT PHOTOCOPY

Complete this Appendix if you have checked the "YES" box under Certification/Requirements on page 1, Form 101. Include activities that will takeplace in CanadaAssessment Act

Title of proposal

Name of applicant's organization

Name of other participating organizations (if applicable)

Name of Location (Please complete an additional copy of Appendix A for EACH location at which research will be undertaken.)

(see the "Requirements for Certain Types of Research" in the NSERC

1. Main characteristics of the location (i.e., physical description & coordinates)

INRS


1) Marine sites :1,2,3,4,5,6) Freshwater sites: 7,8,9,10,11INRS, UQAM, U PEI, MPO, MAPAQ, EC, CEAEQ, Biodome, Aquarium du Québec, Makivic Corporation, Lab_Bell Inc.,IPEV

Baie de Mitis

Page 1 of 33

Family name of applicant Given name Initial(s) of all given names Personal identification no. (PIN)

Michel MFournier

This site has been the object of an intensive scrutiny by our group in last two decades. The Bay is well preserved from any industrial activity being bordered by research institution such as Maurice-Lamontagne Institute and Conservation Park (Parc de la Rivière Mitis, Mitis Garden). The main group of mussels is not accessible to public. The site represents a reference site (source of organisms, sediments, etc.) for many experiments performed by members of our team. Laboratory facilities, provided by Parc de la Rivière Mitis areavailable for the research team, directly on the shore.

only(Total Appendix A

Program Guide for Professors).

APPENDICES A, PLEASE CONTACT NSERC'S ENVIRONMENTAL ASSESSMENT UNIT BY TELEPHONE AT (613) 992-3612 OR


Continue on page 3 of this Form (if necessary).

and/or abroad.

Please ensure that all Appendix A pages are numbered consecutively

This information will assist NSERC in determining whether a screening is required under the Canadian Environmental

Form 101, Appendix A (2011 W)

14327Valid

APPENDIX A (Form 101) CONTINUED

Family name of applicant Page 2 of 33

PROTECTED WHEN COMPLETED Version française disponible

)


DO NOT PHOTOCOPYPersonal identification no. (PIN)

(Total Appendix A only14327 Fournier

Collection of M. edulis2. Principal activity(ies) and activity component(s).


No impacts expected


3. For each principal activity and activity component, list the environmental elements affected and provide a description of those effects.

No impacts expected4. Mitigation measures.



APPENDIX A (Form 101) CONTINUED ADDITIONAL INFORMATION

Page 3 of 33

Use this page to enter additional text from sections 1, 2, 3, and/or 4 (if necessary).

Form 101, Appendix A (2011 W) PROTECTED WHEN COMPLETED Version française disponible

No impacts expected

14327Family name of applicant

FournierPersonal identification no. (PIN)

(Total Appendix A only )


DO NOT PHOTOCOPY








DO NOT PHOTOCOPY


Title of proposal






INRS



Baie des Chaleurs

Page 4 of 33


Michel MFournier

The Baie des Chaleurs watershed is located in Northeastern New Brunswick and the Gaspésie peninsula in Quebec. It is a rich and diverse fresh and marine ecosystem with important river systems, such as the Restigouche and the Nepisiguit. It is one of the most important estuaries draining into the Gulf of St.Lawrence.It covers 1400 km2 and the economy of the Baie des Chaleurs is based on fishery, tourism, forestry and heavy industries. Baie des Chaleurs region is dealing with important erosion problems and sea level rise due to climate change. Direct effects of climate change will occur due to changing patterns of precipitation, longer ice-free season and rising sea level, which will affect hydrology and water quality. In addition, the Baie des Chaleurs region is now dealing with closing industries and a legacy of industrial contaminants [104].These contaminants can be re-circulated to the aquatic environment by erosion of the coastline. Impact of these contaminants on the water resource is now being studied, but adaptation and management strategies are severely needed.






and/or abroad.




14327Valid




)




Collection of M edulis2. Principal activity(ies) and activity component(s).


No impacts expected







Page 6 of 33



No impacts expected





DO NOT PHOTOCOPY








DO NOT PHOTOCOPY


Title of proposal






INRS



Canal Soulanges

Page 7 of 33


Michel MFournier

Canal Soulanges in the St. Lawrence River is a relatively pristine site with very dense populations of zebra mussels. Located upstream of Montreal Island, this site offers riverine conditions, typically observed in the St.Lawrence River, in terms of temperature and other basic water quality parameters. It will therefore represents our reference site for D. polymorpha.






and/or abroad.




14327Valid




)




Collection of D polymorpha2. Principal activity(ies) and activity component(s).


No impacts expected







Page 9 of 33



No impacts expected





DO NOT PHOTOCOPY








DO NOT PHOTOCOPY


Title of proposal






INRS



Kerguelen Archipelago

Page 10 of 33


Michel MFournier

The Kerguelen Archipelago, part of the French Austral and Antarctic Territories, is located at the northern limit of the Southern Ocean. The Polar front runs in the North of archipelago. Moreover, the circumpolar stream arrives in the Western coasts. The associated important water stream is divided in two parts creating distinct hydrological areas in Kerguelen's coastal zone. Such hydrological differences between West and East are also related to major freshwater discharges by rivers in Northern and Eastern part of archipelago. Moreover, water temperature is approximately +6°C in North and +4.5°C for the Southern coasts of KerguelenIslands. In addition of such physicochemical characteristics, high cadmium accumulations were observed in several marine organisms of the Southern Ocean (invertebrates, fishes, birds, mammals...). A recent analysis ofM. edulis desolationis specimens confirmed this Cd contamination. Besides these results on cadmium, Kerguelen's aquatic organisms are contaminated by Persistent Organochlorine Pollutants (i.e. polychlorobiphenyls) in relation to atmospheric transports. Thus, by its strategically geographical position, Kerguelen mussel populations may constitute major sentinels of the effects of global change (ice smelt, water pollution, variations in water temperature and salinity...) on aquatic ecosystems.






and/or abroad.




14327Valid




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No impacts expected







Page 12 of 33



No impacts expected





DO NOT PHOTOCOPY








DO NOT PHOTOCOPY


Title of proposal






INRS



Kuujjuak area

Page 13 of 33


Michel MFournier

The coastline of the Nunavik Region of Northern Quebec encompasses the East Hudson Bay coast, the south coast of the Hudson Strait and the Ungava Bay. The 14 Inuit communities along this coast line all have accessible blue mussel beds where Inuit harvest the shellfish in both winter and summer as part of their country food diet. Largely, all the mussel beds are located in a remote and pristine sub-arctic environment, a distance away from communities, and are free from any point source of contamination. In consideration of logistics, year-round accessibility and feasibility of transporting live samples to the laboratory of Nunavik Research Centre in Kuujjuaq, mussel beds near the communities of Kangiqsujuaq, Kangiqsualujjuaq and Kuujjuaq are suitable uncontaminated sampling sites for this study. At the opposite, the mussels bed in Deception Bay near the community of Salluit, opposite the Hudson Strait, where active nickel mining has beenoperational for the last three decades at the nearby Raglan mine. Elevated levels of various heavy metals including nickel, chromium, cadmium and zinc had been detected in the soft tissues of blue mussels collected from the Deception Bay mussel bed. Climate change, which is happening very rapidly in the North, has an important implication on water quality as well as a potential direct influence on the physiology of blue mussels.






and/or abroad.




14327Valid




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No impacts expected







Page 15 of 33



No impacts expected





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DO NOT PHOTOCOPY


Title of proposal






INRS



Lake Erie

Page 16 of 33


Michel MFournier

where temperature and other environmental conditions (pH, calcium levels, alkalinity) are optimal for zebramussel growth and reproduction. Specimens will be collected from sites subjected to minimal contaminationinfluence.(Total Appendix A onlyProgram Guide for Professors.)APPENDICES A, PLEASE CONTACT NSERC'S ENVIRONMENTAL ASSESSMENT UNIT BY TELEPHONE AT (613) 992-3612 ORProgram Guide for Professors.)Continue






and/or abroad.




14327Valid




)






No impacts expected







Page 18 of 33



No impacts expected





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DO NOT PHOTOCOPY


Title of proposal






INRS



Montreal harbor

Page 19 of 33


Michel MFournier

Montreal Harbor on St. Lawrence River is the most important harbor in Eastern Canada and represents ahighly perturbed and stressed environment with high levels of contamination, both in sediments and in biotaliving in the vicinity of the harbor. The harbor also receives wastewater overflow from municipal effluents.Zebra mussels are found in high abundance along the harbor piers and thus exposed to various contamination stressors. The harbor waters are typically representative of the St. Lawrence River in terms of temperature regime, pH, alkalinity and calcium levels and relatively high speed current ,which may provide lesser growth conditions to zebra mussels.






and/or abroad.




14327Valid




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No impacts expected







Page 21 of 33



No impacts expected





DO NOT PHOTOCOPY








DO NOT PHOTOCOPY


Title of proposal






INRS



Prince Edward Island

Page 22 of 33


Michel MFournier

PEI has a total land area of 566,560 hectares, of which 44% is used for agriculture (PEI DFAF 2008).Although PEI is the smallest province in Canada, it ranks first in terms of potato production. This agriculture requires the use of high quantities of agrichemicals such as pesticides and nitrate-based fertilizers (application rates estimated at 220 kg/ha/yr. of fertilizer; Government of PEI 2008). It was evaluated that PEI is the province with the most intensive use of pesticides in Canada, with total pesticide sales in 2000 of over one million kilogram [105]. The contaminated river waters flow out into the sea surrounding the Island, and then expose the marine invertebrates inhabiting the intertidal zone to the contaminants. In addition, reduced flushing allows accumulation of nutrients thus enabling a high proliferation of macroalgae, which results in a general decrease in the oxygenation rates of seawater in particular under high temperature conditions. Consequently, high prevalence of pathologies was recorded among aquatic invertebrates in the area surrounding the Island. The main example is the development of disseminated neoplasia diagnosed in bivalve mollusks present in North River and Barbara Weit. North River , located in Queens County in the central portion of Prince Edward Island, flows into Charlottetown Harbour and Barbara Weit flows North West into Malpeque Bay. An intensive monitoring program has been put in place since 2005 to follow health of soft-shell clams. To date, this program has shown a high prevalence of disseminated neoplasia in comparison to sites from neighbour provinces such as New Brunswick and Quebec [95].






and/or abroad.




14327Valid




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Page 24 of 33



No impacts expected





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DO NOT PHOTOCOPY


Title of proposal






INRS



Quebec city

Page 25 of 33


Michel MFournier

Quebec City represents actually the northern limit of zebra mussel distribution in the St. Lawrence River and in North America. This site located in the fluvial estuary is characterized by more severe growing conditions for zebra mussels, due to high-speed flow and tidal influence. Water temperature is on average 2oC lower thanthat measured in the St. Lawrence River, thus reducing the growth season for zebra mussels. Due to high flow,rapid water exchange and high turbidity of waters, feeding of zebra mussels is considered being not optimal at that site. Moreover, climate change scenarios predict that Quebec City waters should have major impacts, because of temperature increase and possible salinity increase due to reduced flow rate (river discharge) and saltwater intrusion from estuary. Effects on zebra mussel's populations at this site are difficult to predict.






and/or abroad.




14327Valid




)






No impacts expected







Page 27 of 33



No impacts expected





DO NOT PHOTOCOPY








DO NOT PHOTOCOPY


Title of proposal






INRS



Richelieu River

Page 28 of 33


Michel MFournier

Richelieu River is a tributary of St. Lawrence River and was recently invaded by zebra mussel coming fromLake Champlain [102]. The river is characterized by temperature regime similar to the St. Lawrence River but by low calcium levels (15-18 mg/L) which may become limiting to mussels shell growth. Specimens will be collected at two sites in the upper Richelieu River (St-Paul de l'Île-aux-Noix and Chambly). The St-Paul site islocated upstream of major industrial cities along the Richelieu River and will represent less contaminated conditions, while the Chambly site, located downstream of urban input, will provide zebra mussels having been exposed to multiple sources and high levels of contamination.






and/or abroad.




14327Valid




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No impacts expected







Page 30 of 33



No impacts expected





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DO NOT PHOTOCOPY


Title of proposal






INRS



Saguenay River

Page 31 of 33


Michel MFournier

The Saguenay fjord (Quebec, Canada) has suffered of an important polyaromatic hydrocarbon (PAH) andmetal contamination since the 1950's. The fjord was also part of a network devoted to monitoring of bacterial quality of coastal zones in Eastern Quebec, under the supervision of Environment Canada. This site is under investigation by a research group involving many of the present collaborators since 1995. Work performed with Mya arenaria including evaluation of the physiological fitness, immune, endocrine and reproductive competence has been the object of numerous publications [5,6]. In the last three years, M. edulis has also been the object of research, especially at the mouth of the Saguenay River where salinity is at limit of habitat for theblue mussels.






and/or abroad.




14327Valid




)






No impacts expected







Page 33 of 33



No impacts expected





DO NOT PHOTOCOPY

yes (Y), no (N) or unknown (U) by checking the appropriate box for EACH of the listed activities.Applicants are responsible for verifying whether permits are required for any of the activities listed below. Please indicate

DESCRIPTION OF ACTIVITY

Version française disponibleForm 101, Appendix B (2011 W) Page 1 of 2 PROTECTED WHEN COMPLETED

take place in Canada Complete this Appendix if you have checked the "YES" box under Certification/Requirements on page 1, Form 101. Include activities that will

Family name of applicant Given name


Initial(s) of all given names Personal identification no. (PIN)

Y N UPart 1. - Determination of Physical Work under the CEAA

APPENDIX B (Form 101)Canadian Environmental Assessment Act

Pre-Screening Checklist


DO NOT PHOTOCOPY

Fournier Michel M

Environmental Assessment Act

Part 2. - Determination of Assessable Activities under the CEAA

construction, operation, modification, decommissioning, abandonment or otherin relation to a built structure that has a fixed location and is not intended to be moved frequently?

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

Does any phase of the proposal involve the activity

Activity takes place in a National Park or National Nature Reserve in Canada

Activity takes place on First Nation lands

Activity takes place in the North (Yukon, Nunavut, or the Northwest Territories)

Activity takes place in or within 30 metres of the right-of-way of a power line, a natural gas line, or a railway line

Activity takes place in or adjacent to a water body, resulting in harmful alteration, disruption or destruction of fish habitat (includingthe removal or damaging of aquatic vegetation)

Destruction of fish other than by fishing

Sampling or prospecting for ores or minerals

Disposal of a prescribed nuclear substance other than in a laboratory equipped for such disposal

Deposit of a deleterious or other substance into the environment (in the earth, air, or water)

Any kind of remediation of contaminated land

Deposit of oil, oil wastes or any other substances harmful to migratory birds in waters or in areas frequented by migratory birds

Killing or removal of migratory birds, their nests, eggs, or carcasses or other physical activities that may require a permit or otherauthorisation under the

The removal or damaging of vegetation and/or the carrying on of agricultural activities or the disturbance or removal of soil in a wildlifearea that requires a permit under section 4 of the

Physical activities that are carried on in Canada and that are intended to threaten the continued existence of a biological population in anecodistrict, either directly or through the alteration of its habitat

X Establishment or operation of a field camp in a single location that will be used for 200 person-days or more within a calendar year

Seismic surveying involving more than 50 kg of chemical explosive in a single blast; or marine or freshwater seismic surveying, if duringthe survey the air pressure measured at a distance of one metre from the source would be greater than 275.79 kPa (40 lbs/sq in)X

INRS

Migratory Birds Regulations or Migratory Bird Sanctuary Regulations

Wildlife Area Regulations under the Canada Wildlife Act

and/or abroad. This information will assist NSERC in determining whether a screening is required under the Canadian(see the "Requirements for Certain Types of Research" in the NSERC Program Guide for Professors).

14327Valid


APPENDIX B (Form 101) continued

PROTECTED WHEN COMPLETEDForm 101, Appendix B (2011 W) Page 2 of 2


DO NOT PHOTOCOPY2


Are any authorizations, permits, or licences required to undertake any activity for any phase of the proposal? Ifwith the name of the issuing agency(ies). If

yes, list them below, alongno, please state "None required" and submit this page with the rest of your proposal.

Permit to sample M edulis in sites 1,3,4,5 : Fisheries and Ocean CanadaPermit to sample M edulis in site 2: Parc maritime du SaguenayPermit to sample M edulis in site 6 : requestied by the French teamPermit to sample D. polymorpha in all sites : Environnement Canada

Fournier14327Valid


DO NOT PHOTOCOPY

Date

2012/04/01Family name of applicant Given name Initial(s) of all given names Personal identification no. (PIN)

Fournier Michel MTitle of proposalInteraction between pollution and climate changes: development of improved monitoring strategy

3rd committee reviewer Personal identification no. (PIN)

2nd committee reviewer Personal identification no. (PIN)

NSERC reviewing committee 1st committee reviewer Personal identification no. (PIN)

EArea(s) of expertise

5

DArea(s) of expertise

4

CArea(s) of expertise

3

BArea(s) of expertise

2

AArea(s) of expertise

1hydrology, geographicsBorsellino (Marcelo)

Comission Regional del Rio Bermejo, Sede Centrale,Comandante Fernandez 755Presidencia RoqueSaenz Pena ARGENTINA54 (373) [email protected]

climate change, hydrology

Girard (Pierre)

Centro de Pesquisa do PantanalRua 9, no 305 Boa Esperança

Cuiaba, Mato Grosso BRAZIL55 (653) [email protected]

environmental healthGuimares (Jean Remy Davee)Lab de TracadoresInst. de Biofisica Carlos Chaga Filho/UFRJ,Bloco G, CCS, Iilha do Fundao

Rio de Janeiro BRAZIL55 (212) 561-5339

shellfish health, shellfish immunology, diagnostic

Allam (Bassem)School of Marine and Atmospheric SciencesStoney Brook UniversityNY 11794

Long Island, NY UNITED STATES 1179450001 (613) [email protected]

brackfish water assessment, integrated monitoring

Lehtonen (Kari K.)Department of Biological OceanographyFinnish Institute of Marine ResearchPOB 2,FI-00561Helsinki FINLAND358 (613) [email protected]

PIN

PIN

PIN

PIN

PIN Lang.

Lang.

Lang.

Lang.

Lang.

PROTECTED WHEN COMPLETED Version française disponibleForm 101, Appendix C (2011 W)

Complete Appendix C for all types of grants (except Discovery Grants, Research Tools and Instruments -Category 1, Major Resources Support Grants and Partnership Workshops Program). Read the instructions beforecompleting the appendix.

APPENDIX C Referee Suggestions

(Form 101)

14327Valid

environmental contamination and toxicology, biomarker

Cajaraville (Miren P.)Laboratory of celle biology and histologyUniversity of the Basque CountrySarriena z/g, 48940 Leioa

Basque Country SPAIN

[email protected]

immunotoxicology, pathology

De Guise (Sylvain)Department of PathologyUniversity of Connecticut61 North Eagleville RoadU-3089Storrs, CT UNITED STATES 062691 (860) [email protected]


DO NOT PHOTOCOPY

Date

2012/04/01Family name of applicant Given name Initial(s) of all given names Personal identification no. (PIN)

Fournier Michel MTitle of proposalInteraction between pollution and climate changes: development of improved monitoring strategy

3rd committee reviewer Personal identification no. (PIN)

2nd committee reviewer Personal identification no. (PIN)

NSERC reviewing committee 1st committee reviewer Personal identification no. (PIN)

GArea(s) of expertise

7

FArea(s) of expertise

6

PIN

PIN Lang.

Lang.

PROTECTED WHEN COMPLETED Version française disponibleForm 101, Appendix C (2011 W) Page 2 of 2

Complete Appendix C for all types of grants (except Discovery Grants, Research Tools and Instruments -Category 1, Major Resources Support Grants and Partnership Workshops Program). Read the instructions before completing the appendix.

APPENDIX C Referee Suggestions CONTINUED

(Form 101)

14327Valid