country foods health benefits in a changing canadian arctic · fluctuations of marine food in fat...

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Country FoodsÉ. Dewailly

ArcticNet Annual Research Compendium (2013-14)

Country Foods Health Benefits in a Changing Canadian Arctic

Project LeaderÉric Dewailly (Université Laval)

Network InvestigatorsMichel Lucas (Centre de Recherche du Centre hospitalier de l’Université Laval); André Marette (Institut des nutraceutiques et des aliments fonctionnels (INAF), Université Laval); Alain Cuerrier (Université de Montréal); Pierre Ayotte, Pierre Julien (Université Laval)

CollaboratorsMinnie Grey (Makivik Corporation); Peter Bjerregaard (National Institute of Public Health - Denmark); Amélie Bouchard, Serge Déry, Marie-Josee Gauthier, Elena Labranche (Nunavik Regional Board of Health and Social Services); Kue Young (University of Toronto)

Post-Doctoral FellowsMélanie Lemire (Centre de Recherche du Centre hospitalier de l’Université Laval); Cory Harris (Centre for Indigenous Peoples’ Nutrition and Environment)

Northern HQPMichael Kwan (Makivik Corporation)

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Abstract

To survive in the Arctic, Inuit had for centuries to rely on fish, mammals and some plants such as wild berries and seaweeds. However, since the 1990’s, the consumption of country food has decreased markedly, and the rapid food transition towards a western diet has led to excessive intake of sugar, salt and trans-fatty acids. Global environmental changes also disproportionally affect Inuit dietary patterns in many ways including the availability of local animal and plant species and/or the concentration of environmental contaminants. Once thought to be protected from diabetes and cardiovascular diseases, the current dietary and lifestyle transitions combined to the severe food insecurity context occurring in the Inuit population of Nunavik may change the situation in the near future. The traditional country food diet in Nunavik is very rich in key protective nutrients such as omega-3 polyunsaturated fatty acids and selenium. Wild berries, seaweed and other plants found in Nunavik may also provide plant-derived nutrients and secondary metabolites that also offer unique potential for the prevention or treatment of metabolic disease and associated cardiovascular complications and to offset some deleterious effects of environmental contaminants exposures. With a better understanding of the overall benefits of nutrients present in the different country foods consumed in Nunavik, we can better develop community-based interventions and public policies aiming at improving country food consumption and food security, promote Inuit culture and youth empowerment, minimize the risks from environmental contaminant exposure and the emergence of obesity, diabetes and cardiovascular diseases in this population and across the Arctic.

Key Messages

Our preliminary data on nutrients benefits in country foods show that:

• Beluga mattaaq and nikku (air-dried meat), walrus meat, marine mammal and caribou organs,

and fish eggs are exceptionally high in selenium (> 1 ug/g); beluga and seal meat, blue mussels and marine fish species are also good dietary sources of selenium (> 0.2 ug/g);

• Wild berries in Nunavik display among the highest polyphenols concentrations found in fruit. A tremendous richness and diversity of phenolic compounds are present in the five berry species despite significant intraspecific variation between geographically distinct samples. The berries are also low in simple sugar, fat, and salt yet offer a good source of micronutrients (e.g. Cr, Mn) with very low risk of heavy metal contamination.

• The animal study (complementary study) shows a mild but consistent effect of wild berries on weight loss and significant beneficial effects of cloudberries on insulin sensitivity, of bearberries on intestine integrity, and of both berries on triglycerides accumulation in the liver.

Our first outcomes with the community-based intervention are:

• In February 2013, seeking to translate the combined traditional Inuit knowledge and scientific support for wild berries into a community-kitchen initiative, we developed the “Purple Tongue Project” with Individual Path Learning (IPL) students in Tasiujaq and Kangiqsualujjuaq schools. This pilot-project is a new form of research-milieu collaboration with not-for-profit partners (Biopterre, Institut de technologie agroalimentaire), the Nunavik Regional Board of Health and Social Services and the Kativik School Board. Our objectives with this intervention are to mobilize TK and scientific support for wild berries, to improve their consumption and availability throughout the year, propose attractive and local healthy options to sweet beverages and snacks, and stimulate youth empowerment and employment. Two workshops with IPLs were conducted: in September in Tasiujaq and in November in Kangiqsualujjuaq. Along with berry picking, we conducted activities

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about TK of plants, their nutritional benefits and cooking berry products. We also developed a booklet with the berry recipes and other informations covered during the activities.

• The activities of the project, co-funded by the Kativik Regional Government (KRG), the NRBHSS and Air Inuit, had a great success. In September and October 2012, the IPL students collected up to 40kg of berries for the project. Nathalie Ross, Tasiujaq school director, mentioned: “For the first time, my IPL students were the ones having the greatest and funkiest project in the school. We broke a taboo that IPL students are just drop-off and not good for anything.” As reflected by student engagement and community feedback, investing in community initiatives to empower youth while generating social economic opportunities is invaluable to effective knowledge sharing and educational outreach. The next phase of the project is now in preparation.

Objectives

We will measure new biomarkers of Se status and selected phytochemicals, and investigate relations between country food consumption, nutrient biomarkers and emerging health issues such as obesity, diabetes and cardiovascular diseases risk factors in Inuit adults. We will also collect country foods and measure total Se, identify the forms of Se and measure other micro- and macronutrients (vitamins, minerals, fibres, proteins, lipids and phytochemicals) present in various traditional foods prepared and eaten by Inuit populations. We will also explore the traditional knowledge, health benefits (obesity, diabetes, cardiovascular health, and immune system), social and gender role, and economic aspects of different country foods found in Nunavik via a literature review and discussions with community stakeholders in different villages. From this preliminary research on health benefits of country foods, we will develop and pilot a community-based intervention project to eventually

expand in scale and scope. Building on this successful collaboration, we will submit a more comprehensive and interdisciplinary research/intervention proposal to the 2014 “Pathways to Heath Equity for Aboriginal People” grant competition administered by the Institute of Aboriginal Peoples’ Health (CIHR).

Specific objectives

1. Studies among the Inuit adult population from Nunavik:

1.1. Measure various new biomarkers of Se status [selenoproteins such as selenoprotein P (SelP) and glutathione peroxidases (GPx1, 2, 4), plasma total Se (P-Se), inorganic Se (Se+4 and Se+6), selenoneine, selenocysteine (SeCys), selenomethionine (SeMet) and selenoalbumin (SeAlb)] in archived blood samples from the 2004 Nunavik Inuit Health Survey (NIHS) (n=900) randomly selected adults, between 18 and 74 years old); (Status: to be completed in 2014-2015)

1.2. Investigate the associations between country food intakes data from the NIHS and the various new biomarkers of Se status as well as the different associations among Se biomarkers; (Status: to be completed in 2014-2015)

1.3. Examine the associations between the biomarkers of Se status and diabetes and CVD risk factors (e.g. fasting glucose and insulin, lipids profile, blood pressure, PON1 activity, inflammatory biomarkers) evaluated during the NIHS, taking into account co-variables, interactions with contaminants (MeHg, PCBs, other metals) and other potential confounders; (Status: to be completed in 2014-2015)

1.4. Examine the association between wild berry and seaweed consumption and obesity, diabetes and CVD risk factors in the NIHS; taking into account co-variables,

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interactions with contaminants (MeHg, PCBs, other metals) and other potential confounders; (status: to be completed in 2014-2015)

1.5. Measure selected phytochemicals (flavonoids, carotenoids, etc.) and their metabolites in archived blood samples from the NIHS focusing on a subsample of the study participants reporting high or low wild berry and seaweed consumption (n=125) to further explore the associations described in obj. 1.4.; (Status: to be completed before March 2014)

2. Studies involving Nunavik country foods

2.1. Collect selected country foods (raw, dried, aged: marine mammals, freshwater and marine fish, land mammals, wildfowl, wild berries, seaweed and other plants) from several Nunavik villages in collaboration with the Hunter support program, community members and the Nunavik Research Center of Makivik Corporation; (Status: completed)

2.2. Determine the age of mammals, fish, birds and seafood country foods samples; (Status: completed)

2.3. Measure total Se concentrations and Se chemical species in country food samples; (Status: completed with Se chemical species data forthcoming)

2.4. Characterize and quantify different phytochemicals (flavonoids, carotenoids, etc.), macronutrients, vitamins, minerals and environmental contaminants in wild berries, seaweed and other plants consumed by Inuit in Nunavik, and evaluate the effect of berry cooking processes (see obj. 3.1) on these compounds; (Status: completed with macronutrient data and cooking process data forthcoming)

2.5. Assess the bioactivity (evaluate the protective potential) of wild berry and

seaweed extracts in models pertinent to the prevention and treatment of chronic diseases (in vitro antioxidant and anti-inflammatory activities); (Status: completed with anti-inflammatory data forthcoming)

2.6. Conduct a literature review on bioactive phytochemicals, micronutrients and macronutrients possibly found in wild berries, seaweed and other plants species consumed in Nunavik, on nutritional and cardiovascular health benefits, Inuit traditional ecological and medicinal knowledge, social and gender roles associated gathering and consumption as well as environmental and climate change aspects related to these plants in Nunavik and elsewhere. (Status: completed)

2.7. Determine the seasonal variation of n-3 and total lipid content in Arctic char; (Status: half of the sampling conducted, to be completed in 2014-2015)

2.8. Characterise the protein profile of fish species most frequently consumed by Inuit (Arctic char, brook trout, lake whitefish, sculpin, lake trout), ringed seal, blue mussel, caribou, ptarmigan and Canada geese meat in Nunavik (n=50); (Status: to be completed before March 2014)

2.9. Conduct two experimental studies in order to evaluate the anti-diabetic effects of seal oil (misirak or aged seal blubber), arctic Char proteins and seal proteins; (Status: to be conducted in 2014-2015)

3. Studies and intervention projects with commu-nity members

3.1. Discuss with different stakeholders in the communities in order to explore the ecological, health, cultural and socio-economic aspects that could be of interest for the communities and how we could provide information, support and develop community-based intervention aiming

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to better promote the consumption, the distribution and eventually the commercialisation (ex. wild farming etc.) of berries, seaweed and other plants between communities, and if of interest, to other regions outside Nunavik. (Status: on-going within actively collaborating community organizations) [2014-15: scaling up to include additional communities and regional organizations]

3.2. Our Sea, Our Health: Discuss with community members from two villages of the Hudson Strait in Nunavik in order to explore and document the traditional use of marine foods and marine fats for medicinal and other properties, the seasonal fluctuations of marine food in fat content and related consumption as well as their importance for Inuit identity and culture. (Status: to be conducted in 2014-2015)

Introduction

To survive in the Arctic, Inuit had for centuries to rely on fish, mammals and plants such as wild berries. This traditional country food diet is essential in providing social cohesion and cultural continuity (1), and is believed as one of the main factors that protects Inuit from many chronic diseases (2). In Nunavik, northern Quebec, the prevalence of type 2 diabetes (T2D) and cardiovascular diseases (CVD) among Inuit has been low compared to that of general populations (3), and much lower than in most Aboriginal populations which are now struggling with T2D epidemics (4). However, the current dietary and lifestyle transitions combined to the severe food insecurity context occurring in Nunavik may change the situation in the near future (5). Country foods do not always appeal to Inuit youth and their actual consumption of sweet beverages are predictions for major obesity and T2D epidemics (5). Other global changes also disproportionally affect Inuit in many ways, including the availability of local animal and plant species and/or the long-range

transport of environmental contaminants to the Arctic (6).

Traditional Inuit Knowledge (TK) passed down through generations emphasizes the importance of wild animals and plants in the Inuit diet, medicine, and culture (1). Although some traditional marine foods have become primary vectors for MeHg exposure (6), our research endorses TK regarding health benefits of Inuit country foods. Exceptional concentrations of omega-3 (n-3) and selenium (Se) are found in the Inuit diet, and Nunavik populations present some of the highest n-3 and Se status in the world (7). Indeed, several marine fish and mammals are naturally rich in eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, while docosapentaenoic acid (DPA), uncommon in fish oils, is a substantial component of ringed seal and beluga blubbers as well as beluga mattaq, an Inuit delicacy made of beluga skin and blubber (8, 9). However, important seasonal variation in fat content may be found in marine foods. Beluga skin and mattaaq are also known as important sources of Se (8), but other sources may also be present in the Nunavik diet.

High n-3 intakes are associated with a number of health benefits in this population (7, 10-13). There is also evidence that high Se intake may counterbalance some toxic effects of MeHg exposure in adult populations living on fish consumption (7, 14-17). However, Se health benefits with respect to T2D and CVD remain contentious: plasma Se is both negatively and positively related to T2D and CVD risk factors in other populations (18). In addition to plasma Se, the most common biomarker of Se status, several other blood biomarkers (e.g. Se-enzymes, small Se-compounds) can provide additional insights (19, 20). Importantly, the chemical forms of Se found in different foods will influence Se absorption and bioavailability for subsequent selenoprotein synthesis with potential impacts on health (21).

With limited access to fruits and vegetables, wild berries, seaweed and other plants found in Nunavik may also provide otherwise scarce plant-derived

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nutrients and secondary metabolites that not only improve dietary quality but may help prevent or manage T2D and associated CVD complications. In addition to fibre and minerals, many plant-based foods are rich in antioxidants including vitamins C and E as well as polyphenols and carotenoids (22, 23). Several of these phytochemicals may reduce lipid peroxidation and LDL oxidation, improve insulin secretion and sensitivity, or act as vasoprotective and anti-inflammatory agents (22, 23). Beyond these bioactivities, certain phytochemicals also chelate heavy metal ions, potentially reducing the bioavailability and subsequent toxicity of dietary contaminants such as methylmercury (24, 25). Moreover, specific proteins found in marine country food may contribute to beneficial actions on insulin sensitivity, lipid metabolism and inflammation (26). Up to now, little information is available on plant phytochemicals and protein profile of fish species and other country foods in Nunavik.

With a better understanding of the overall benefits of nutrients present in the different country foods consumed in Nunavik, we can better develop community-based interventions aiming to improve country food consumption and food security, promote Inuit culture and youth empowerment, minimize the risks from environmental contaminant exposure and the emergence of obesity, T2D and CVD in this population and across the Arctic.

Activities

New biomarkers of Se analysis (Objective 1.1, 1.2, 1.3)

Blood analyses for Se-enzymes and small Se-compounds are currently being conducted by Dr. Pierre Ayotte research team at the Institut National de Santé Publique du Québec (co-funded by the Northern Contaminant Program (NCP), PI: Dr Ayotte, 2012-2014; 2014-2016 (submitted)). A new collaboration for selenoneine (new small Se-compound) speciation analysis in blood samples and country foods has also

been established with the Laboratoire de Chimie Analytique Bio-inorganique et Environnement (LCABIE) / CNRS – Université de Pau (PA), France in 2013 (PI: Dr Ayotte and Dr Bouyssières, FQRNT-CFQCU, 2013-2015). The complete new biomarkers of Se analysis results are expected for mid-2014. The statistical analyses (objective 1.3.) will be conducted in 2014-2015.

Wild berries and seaweeds health benefits (Objective 1.4 and 1.5)

Phenolic metabolites will be analyzed in a sub-sample of the NIHS study (n=125) at the laboratory of Dr. Yves Desjardins at ULaval by UPLC-MS/MS. The analyses are scheduled for February-March 2014.

Dr Lucas, Dr Lemire and a PhD student are in charge of the statistical analyses aiming to examine the association between wild berry and seaweed consumption and obesity, diabetes (fasting insulin, glucose, etc.) and CVD risk factors (inflammatory biomarkers, etc.) in the NIHS; taking into account co-variables, interactions with contaminants (MeHg, PCBs, other metals) and other potential confounders. These analyses will be conducted in 2014-2015.

Collection of country food samples (Objective 2.1)

Building on our collection of more than 300 land animal, wildfowl, marine mammal, fish, and plant food samples collected by Dr Kwan, Dr Lemire and Ashleigh Downing in 2012, we continued collecting country food samples in 2013.

Selenium total concentration and speciation, and age determination (Objective 2.2 and 2.3)

Total Se concentrations analysis and age determination (for animal samples only) were done by Dr Kwan at the Nunavik Research Center. Almost all samples were analysed for Se and most of the results shown in the Tables 1 and 2 are adjusted for age (results section). More results are expected for caribou nikku (air-dried),

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Table 1. Total Hg and Se results (age-adjusted) in country foods collected in Nunavik between 2008 and 2013

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Atlantic salmon and brook trout eggs over the coming months. Se speciation analysis will also be done by Dr Laurie Chan at Ottawa University over the coming months (co-funded by NCP, 2012-2014).

Wild berries, seaweed and other plants analysis and bioactivity (Objective 2.4 and 2.5)

Dr Harris is currently supervising the measurements of the different micronutrient, macronutrient and phytochemical concentrations in wild berries, seaweed and herbs at McGill, Ottawa University and the Institut des neutraceutiques et aliments fonctionnels (INAF) at Université Laval. Following up on our 2012-13 data

for five species of berries used in our in animal study (complementary study, see below), we extended our comprehensive nutrient and phytochemical analyses to multiple samples of each species collected from various regions and habitats. We similarly evaluated the chemistry and chemical variability of three and four species of seaweeds and herbs, respectively (Figure 1).

More analyses will be conducted over the coming weeks to evaluate the effect of the different cooking processes used in the Purple Tongue recipes (dehydration, low-temperature cooking, high-

Table 2. Hg and Se concentrations (age-adjusted) in country foods from Nunavik and elsewhere in the Arctic (continued)

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temperature sterilisation, etc.) on micronutrient, macronutrient and phytochemical concentrations.

For the bioactivity assessment, Dr Harris and two trainees assessed the in vitro radical scavenging (Yi Yong, MSc, McGill) and anti-inflammatory activity (Brendan Walshe-Roussel, PhD, University of Ottawa) of plant food extracts. The in vitro antioxidant are completed and the anti-inflammatory activities will be conducted over the coming weeks.

In 2013, in parallel to the present study on country food benefits, we conducted a complementary animal study under the supervision of Dr Marette (INAF, ULaval). This in vivo study investigated the effects of five different species above – the types most commonly consumed by the Inuit of Nunavik – on several metabolic and physiological endpoints associated not only with obesity but diabetes and CVD risk as well.

Results of both in vitro and in vivo experiments were then considered relative to the chemical composition of the each extract, all of which contained numerous bioactive compounds with reported health benefits, most notably phenolic metabolites.

Literature review on wild berries, seaweed and other plants (Objective 2.6)

Ashleigh Downing completed a comprehensive literature review and developed a searchable EndNote library spanning that included:

• nutritional value and phytochemistry of Nunavik food plants and closely related species;

• bioactivity, pharmacology, and toxicology of Nunavik food plants species and their metabolites;

• effects of climate change on the abundance, distribution, and contaminant burden of northern plant species;

• community-based interventions addressing food insecurity, chronic disease, and knowledge exchange within indigenous communities in Canada and abroad.

Dr Lemire, together with Dr Cuerrier, Dr Harris and Ashleigh Downing, combined insights from our experimental results and literature review to develop a knowledge sharing poster that was subsequently distributed to community partners and posted in schools and community centres. We similarly used these data and information for the development of our educational outreach and pilot intervention project (obj.3.1).

Seasonal variation of n-3 and total lipid content in Arctic char (Objective 2.7)

During fall (season when the fish are more fat), Dr Kwan collected over 50 samples of Arctic char meat, fat (belly), liver, skin and pitsik (air-dried meat and skin) from the Vachon River in Kangiqsuk. More samples will be collected at the end of the spring (season when the fish are lean) of 2014. Total fat and n-3 analysis will be undertaken over the coming weeks at Dr Julien laboratory at ULaval.

Figure 1. Wild berry, seaweed and other plant species included in the study

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Protein profile in country foods (Objective 2.8)

Country food samples were collected and the analyses are scheduled for the next weeks at Dr. André Marette laboratory.

Community based-intervention: The Purple Tongue Project (Objective 3.1)

In fall 2012, while visiting communities to collect native plants, community members expressed a desire to consume more berries, but also highlighted numerous obstacles, from economic (e.g. time constraints) and social (e.g. gender and generational issues) to physical (e.g. no storage space in freezers) and environmental (e.g. seasonality).

At the KRG meeting in Puvirnituq in December 2012, we were introduced by Helene Raymond, journalist at Radio-Canada, to Maxim Tardif from Biopterre (www.biopterre.com), a Centre collegial de transfert de technologie affiliated to the Institut de Technologie Agricole in La Pocatière, Quebec. Biopterre has a recognize expertise for the development of bioproducts made from non-timber forest products together with small enterprises and Cree municipalities, and in knowledge transfer, capacity building and implementation support to local partners in order to promote economic diversity of rural communities.

In February 2013, seeking to translate the combined traditional Inuit knowledge and scientific support for wild berries into a community-based initiative, we engaged community stakeholders, the NRBHSS, Kativik Regional Government (KRG), Kativik School Board (KSB), and not-for-profit partners (Biopterre) to develop the “Purple Tongue Project” in in Tasiujaq and Kangiqsualujjuaq schools (KRG, 50K; NRBHSS, 16K+in kind, KSB, in kind; Air Inuit, in kind; Biopterre, 50K+in kind; Nasivvik, 5K). With Biopterre, we developed novel wild berry products (baby puree, roll ups, dried berries, granola bars, sorbet and frozen yogurt) to be produced by Individual Path Learning (IPL) students in school.

Our objectives with this intervention project are to improve wild berry consumption, distribution and availability throughout the year, propose attractive, culturally-relevant and local healthy alternatives to soft drinks and snacks, and stimulate youth empowerment and employment.

New formulations of the berry products were first evaluated by community representatives, whose feedback inspired recipe refinement and alternative formats. The berry products had to present low sugar content, produced at low cost, using locally available food items and simple cooking techniques, attractive for the youth, and in accordance with Inuit taste and culture. In Tasiujaq in September 2013, we introduced our selection of berry products to again receive feedback – this time from the IPL students from the two villages. Along with berry picking, we conducted several activities about TK of plants, nutritional benefits of Nunavik berries, and berry patch mapping using GPS.

This first workshop in Tasiujaq with IPL students from the villages was a success. The students were present and attentive, tested the berry products, and participated in the outdoor activities. Everyone enjoyed the activity on TK of the plants with Susie Morgan, an elder from Kangiqsualujjuaq. As Nathalie Ross, Tasiujaq school director, mentioned: “For the first time, my IPL students were the ones having the greatest and funkiest project in the school. We broke a taboo that IPL students are just drop-off and not good for anything.” From this experience, Mrs. Ross suggested that the intervention project could become an integral part of the pedagogical guide of IPL students on a 3 years basis. Collectively, we decided to focus on practical hands-on and tool kit activities (berry mapping and harvest assessment, making a herbarium, cooking berry recipes, hygiene and safety tips in a kitchen, tool construction, the basics of project management) to better involve young men in the project. Prior to our second visit, IPL students had already collected more than 40 kg of berries in preparation for the next phase of the project.

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In addition to implementing the pilot project, we also developed a recipe book and information package both in English and French (English version in Apendix II) for the IPL students and the communities. The Inuktitut translated version is expected for the next month.

The second workshop was held in Kangiqsualujjuaq in November. We introduced the recipe book, conducted an hygiene and safety activity, presented and realized twice each recipe of the berry products with the students, went ice fishing, visit the museum at the Kurrurjuaq National Park, discussed with Kangiqsualujjuaq elders (Tivi Etok, Susie Morgan, Sarah Pasha) about TK. The IPL student had their berry products tasted with preschool children at the Day care and with other high school students in their respective schools.

For the evaluation of the berry products (1st workshop) and the evaluation of the project (2nd workshop) we used interactive voting devices (quantitative questionnaires). Semi-directed interviews are planned with IPL school teachers and school directors are scheduled for the

coming month. A scientific article on the design and preliminary outcomes of the pilot-project is already in preparation.

In the next phase (2014-2015) will aim to: refine berry recipes, develop new recipes with seaweeds, plants and mushrooms, study health, environmental and socio-economic related- issues, and develop a hands-on and educational pedagogical guide for IPL students across Nunavik with KSB and the NRBHSS (scale-up of the pilot-project). An additional collaboration with Nunavik Parks is also envisioned.

More recently, our team received funding from Nasivvik (30K) and applied to a CIHR planning grant (25K). With this funding, we aim to conduct in 2014 a large and intergenerational community consultation campaign in Nunavik villages to develop more partnerships and a collaborative research and capacity-building intervention agenda to address

many issues at once: promote Inuit culture, traditional knowledge and nutritional benefits of Nunavik country foods (including berries and marine diet), improve food security, and empower the youth to generate social economy opportunities, and to minimize MeHg exposure (particularly among women of childbearing age) and the emergence of obesity and diabetes in Nunavik.

Results

Objectives 2.1 – 2.3: Se analysis and age determination in Nunavik country foods (co-funded by the Northern Contaminant Program)

Overall, these results show that:

• Beluga meat (raw and nikku), marine mammal organs and lake trout present very high total Hg concentrations (> 0.5 µg/g), while beluga mattaaq and ringed seal meat are low-intermediate in Hg (0.2 – 0.5 µg/g). Some samples of marine bird eggs presented very high Hg concetrations (>1 µg/g);

• Very high total Hg is found in marine mammal organs, however, their MeHg concentrations are much lower (less than 25%);

• Almost all other country foods are low in Hg (< 0.2 µg/g);

• Beluga mattaaq and nikku, walrus meat, marine mammal and caribou organs, and fish eggs are exceptionally high in total Se (> 1 µg/g);

• Beluga and seal meat, blue mussels and marine fish species are also good dietary sources of Se (> 0.2 µg/g).

Objective 2.4: A. Phytochemical analysis of plant food extracts

We characterized:

• Anthocyanins in berries by HPLC-DAD (Figure 2)

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• Procyanidins by HPLC-fluorescence (Figure 3)

• Flavonoids and phenolic acids by UPLC-MS (Figure 4)

• Ellagitannins in berries by HPLC-DAD (Figure 5)

• Abscisic acid in berries by UPLC-MS (Figure 6)

These analyses demonstrate that:

• All berry species contain a significant quantity and diversity of phenolic metabolites.

» Alpine bearberry, blackberry, and particularly blueberry are rich in flavonoids, specifically quercetin derivatives with established bioactivities; quercetin content in blueberries exceeded that of any food

listed in the USDA Database for Flavonoid Content of Selected Foods (http://www.ars.usda.gov/SP2UserFiles/Place/12354500/Data/Flav/Flav3-1.pdf)

» Alpine bearberry and blackberry are rich in anthocyanins, metabolites often listed and standardized as active constituents in berry-based health products; contents in blackberry rank among the highest listed in the USDA Database for Flavonoid Content of Selected Foods (Note: anthocyanins are a sub-class of flavonoids)

» Redberry, blackberry, and alpine bearberry are rich in procyanidins – higher than found in most fruits (USDA Database for Procyanidin Content of Selected Foods, http://www.ars.usda.gov/SP2UserFiles/

Figure 2. Anthocyanin contents of Nunavik berries collected from different geographical locations. Quantitative results were obtained for >20 anthocyanin structures identified among the five species but data are expressed as total anthocyanins (mg) per 100g fresh weight.

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Figure 3. Procyanidin contents of Nunavik berries (A), seaweeds and herbs (B) collected from different geographical locations. Quantitative results were obtained for monomeric through decameric and larger polymeric structures but data are expressed as total procyanidins (mg) per 100g fresh weight.

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Figure 4. Flavonoid and phenolic acid contents of Nunavik berries (A), seaweeds and herbs (B) collected from different geographical locations. Quantitative results were obtained for up to 18 flavonoids and 34 phenolic acid derivatives monomeric through decameric and larger polymeric structures but data are expressed as total flavonoids/phenolic acids (mg) per 100g fresh weight.

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Place/12354500/Data/PA/PA.pdf); these compounds possess antioxidant and metal chelating potential and also effect gut microflora and gastrointestinal function. Whereas the majority of procyanidins in all species were mononers and small polymers, blueberry, blackberry, and bearberry contained significant amounts of large polymers as well.

» Only cloudberry and alpine bearberry (black and red ssp.) contain ellagitannins

» Blueberry and redberry contain abscisic acid, a plant hormone with putative anti-diabetic effects in humans

• Herbs used for tea are rich in flavonoids and procyanidins, contributing to taste and traditional medicinal properties

• Mountain sorrel, seabeach sandwort, and the seaweeds (plants used for spice and snacking) did not contain large quantities of phenolic metabolites. However, our phytochemical characterization is the first for many of these species.

• Phytochemical variation between different geographical samples of the same species can be significant, up to two fold for individual metabolites, such variability is only observed when concentrations in the species are consistently high relative to other species (e.g. blackberry anthocyanins are extremely variable but always higher than most other species).

Objective 2.4: B. Nutrient analysis of plant food extracts

We characterized:

• Simple sugars in berries by HPLC-DAD (Table 3)

• Major minerals by atomic absorption spectroscopy & trace minerals by ICP-MS (Tables 4-5)

Figure 5. Ellagitannin contents of three Nunavik berry species collected from different geographical locations. No ellagitannins were detected in other species. Quantitative results are expressed as total ellagitannins (mg) per 100g fresh weight.

Figure 6. Abscisic acid contents of Nunavik berry extracts used in the in vivo study. Quantitative results for abscisic acid associated derivatives are combined and expressed in μg per 100g fresh weight.

Table 3. Simple sugar contents of Nunavik berry collections used in our In vivo study. The presented values are the sum of detected glucose and fructose levels expressed in mg per 100g fresh fruit.

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Table 4. Major mineral contents of Nunavik berries collected from different geographical locations. Quantitative results were obtained for P, K, Ca, Mg, N; data for P and Mg presented. Servings calculated as 1 cup of berries (142g)

Table 5. Trace mineral contents of Nunavik berries collected from different geographical locations. Quantitative results were obtained for Cr, Fe, Mn, Zn, Cu, Ni, Se, As, Pb, Cd, Ba, Co, Al, V, Rb, Sr, and Y; data for Cr, Fe, Mn and Cu are presented. Servings calculated as 1 cup of berries (142g).

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• Vitamins C, E, and A (including beta-carotene) by HPLC-DAD (Table 6)

• Total protein, total fat, soluble and insoluble fibres (data forthcoming)

» Note: fatty acid profiles of the five berry species were completed in 2012

These analyses demonstrate that:

• Fresh wild Nunavik berries:

» Are low in sugar (3-5g/100g), low in fat (0.6-0.9g/100g; 2012 data), low in salt (< 4 mg/100g);

» Are very good sources of Cr, Mn & Cu;

» Contain low-to-moderate amounts of Mg, Fe and vitamins A, C & E;

» Contain only trace levels of potentially toxic metals (As, Pb, Cd, Hg) (data not shown but confirmed by ICP-MS analyses at both McGill (Dr. Harris) and Nunavik Research Centre (Dr. Michael Kwan). One blackberry sample (of >30 total samples) was high in a particular heavy metal (Cd)

» Are safe and nutritious!

• On a per gram basis, seaweeds are lower than berries in sugar and vitamins C & E but higher in minerals and heavy metals; herbs are generally higher in both vitamins and minerals (data not shown). Because the plants are – to our knowledge - rarely consumed in large amounts or steeped (rather than eaten), their dietary contributions are likely lower than those of berries. More research into how and when these foods are consumed is required.

Objective 2.5: A. In vitro bioactivity of wild berry extracts

We characterized:

• Antioxidant activity (in vitro) using the Oxygen Radical Absorbance Assay (ORAC) (Figure 7)

• Anti-inflammatory activity (in vitro) using tumour necrosis factor alpha (TNF-alpha) ELISA and nitric oxide release assays in a monocytic cell line. (Data forthcoming)

Table 6. Contents of vitamins C, E, and A in Nunavik berries collected from different geographical locations. Servings calculated as 1 cup of berries (142g).

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Objective 2.5: B. Wild berry effects using a diet-induced mouse model of obesity

(complementary study with funding support from Dr Marette and Lucas)

Following an 8-week study on the effects of five individual berry extracts on the development of diet-induced obesity in mice, our results demonstrated that:

• All extracts except blueberry reduced body weight (mild effect)

• Cloudberry extract increased insulin sensitivity

• Cloudberry & bearberry extracts reduced lipid accumulation in the liver (fatty liver) (Figure 8)

• Bearberry improved Intestine integrity

Figure 7. Representative oxygen radical scanvenging capacity (ORAC) values of extracts from five berry species (A). Activity varied within and between species but antioxidant activity was strongly correlated increasing total phenolic (B) and procyanidin (C) content.

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These two set of analyses demonstrate that:

1. All berry species exhibited one or more bioactivity relevant to the treatment or prevention of obesity, diabetes, and CVD:

• Berries that are richest in phenolic metabolites (blackberry, bearberry, redberry) offer the greatest antioxidant potential; issues of bioavailability require additional research to determine the significance in vivo;

• Berry extracts appear to elicit weak-to-moderate anti-inflammatory effects on immune-stimulated monocytes (preliminary data not shown);

• Although no extract completely protected mice from diet-induced obesity, most extracts reduced at least one important pathophysiological

parameter (e.g. insulin resistance, fatty liver, weight gain)

» Reduced fatty liver was observed only in the two species containing ellagitannins – potential mechanistic insight

» The multiple positive effects of cloudberry and bearberry warrant further investigation

2. Based only on in vitro data (not shown) the two Labrador tea species possessed greater antioxidant capacity than mountain sorrel and berries, which demonstrated similar activity. Seabeach sandworth and the seaweed species possessed only weak activity. Forthcoming results about non-berry anti-inflammatory activity will add insight into the potential health benefits of these plant foods.

Objective 2.7 - The Purple Tongue Project (Figure 9.)

See Activity section for more details.

Discussion

Most of these results are preliminary and will be integrated over the coming months. More results are expected soon (described in the 2013-2014 Activity section for the on-going projects, and Planned activities section for the new objectives). This will

Figure 8. Effects of high fat – high sugar (HFHS) diet on liver triglycerides (TG) and the effects of berry extracts on HFHS induced fatty liver. (n= 6, * p< 0.05 vs. CHOW; # p< 0.05 vs. HFHS)

Figure 9. Berry products developed in collaboration with Biopterre and consultation with community partners (after three rounds of feedback and recipe optimization).

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help us developing a global understanding of the overall benefits of country foods in Nunavik.

Conclusion

Better understanding the overall benefits of country foods consumed in Nunavik will support and orient community-based projects and public policies aiming to improve food security, promote Inuit culture and youth empowerment, minimize the risks from environmental contaminant exposure, and the emergence of obesity, diabetes and cardiovascular diseases in this population and across the Arctic.

Acknowledgements

We are thankful to the Nunavik Nutrition and Health committee, the Nunavik Regional Board of Health and Social Services, the Kativik Regional Government, the Kativik School Board, the Nunavik Research Centre for their support and participation to our projects. Dr Lemire is by a Banting-CIHR fellowship (2012-2014) and Dr Harris is also supported by a Banting-CIHR fellowship (2011-2013).

We are grateful to all Nunavimmiut who participated to the elaboration and implementation of the Purple Tongue Project. We deeply thank Nathalie Ross, Nancy Etok, Mark Brazau, Michelle Hallé, Daniel and Annie Annanack, Margaret, Susie Morgan, Sarah Pasha, Tivi Etok , France Brind’Amour, and Charlie Munick who readily jumped in with us on this adventure of developing products for Nunavik youth while having fun and learning together. A second special thanks to IPL students from Tasiujaq and Kangiqsualujjuaq who joined the project. They are inspiring.

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Publications

(All ArcticNet refereed publications are available on the ASTIS website (http://www.aina.ucalgary.ca/arcticnet/).

2014-01-22: Dewailly E, Ayotte P, Muckle G, Riva M, Lucas M, Lemire M, “Twenty-five years of scientific progress on contaminants and health in the Arctic”, 8th Arctic Frontiers conference, Norway, Tromso

2013-12-11: Lemire M, Ayotte P, Chan L, Dewailly E, Achouba A, Dumas P, Laird B, Vanié P, Kwan M, “Country foods and cardiovascular health in Nunavik: studying the complex balance between selenium and environmental contaminants”, ArcticNet’s 9th Annual Scientific Meeting, Canada, Nova Scotia, Halifax

2013-12-11: Harris C, Lemire M, Lucas M, Kwan M, Cuerrier A, Ayotte P, Desjardins Y, Marette A, Dudonné S, Pilon G, Gauthier MJ, Bouchard A, Labranche E, Déry S, Grey M, Dewailly E, “Health benefits of wild berries, seaweeds and other plants in Nunavik”, ArcticNet’s 9th Annual Scientific Meeting, Canada, Nova Scotia, Halifax

2013-11-25: Lemire M, Tardif M, Cuerrier A, “Wild berries, seaweeds and other plant health benefits, Research & Intervention project”, Kativik Ragional Government annual council, Canada, Quebec, Kuujjuaq

2013-10-10 : Lemire M “Les bénéfices et les risques de l’alimentation traditionnelle au Nunavik: les liens indissociables entre la santé, l’environnement et la culture Inuit”, Ministère du Développement Durable, Environnement, Faune et Parcs., Canada, Quebec, Quebec

2013-09-25: Lemire M, Harris CS, Lucas M, Kwan M, Cuerrier A, Ayotte P, Marette A, Tardif M, Gauthier MJ, Bouchard A, Labranche E, Déry S, Grey M, Dewailly E, “Wild berries, plants and seaweeds health benefits in a changing Canadian Arctic”, Northern Contaminants Program, 20th annual Results Workshop, Canada, Ontario, Ottawa

2013-06-06 : Lemire M, “Les bénéfices et les risques de l’alimentation traditionnelle au Nunavik: les liens indissociables entre la santé, l’environnement et la culture Inuit”, 17e Colloque du Chapitre Saint-Laurent, Canada, Quebec, Montreal

2013-06-05 : Lemire M, “Les bénéfices et les risques de l’alimentation traditionnelle au Nunavik”, Écosystèmes et enjeux globaux en santé publique vétérinaire (graduate course SPV6530), Faculté de médecine vétérinaire, Université de Montréal, Canada, Quebec, Saint-Hyacinthe

country foods health benefits in a changing canadian arctic · fluctuations of marine food in fat...

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