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    Ageing, cognition and omega-3 fatty acids : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Psychology at Massey University, Wellington, New Zealand
    (Massey University, 2018) Mengelberg, Alexia
    The evidence for omega-3 polyunsaturated fatty acids (n-3 PUFA) in fish oil supplements being a safe therapeutic agent is steadily growing. However, there is still a lack of evidence around the cognitive effects of n-3 PUFAs in older adults with Mild Cognitive Impairment (MCI), the moderating effect of the Apolipoprotein E (APOE) ɛ4 allele on cognition and well-being, and the popularity of fish oil supplements in New Zealand (NZ). The primary aim of this thesis was to conduct a clinical trial to investigate the cognitive effects of a high dose docosahexaenoic acid (DHA) fish oil supplement in older adults with MCI, and to examine how the presence of the APOE ɛ4 allele affects the efficacy of fish oil. The trial involved a 12-month randomised, double-blind, placebo-controlled design with testing sessions at baseline, 6-months, 9-months and at the end of the trial. Seventy-two adults with MCI between the ages of 60 and 90 were recruited from Wellington, NZ. Participants were either given a DHA fish oil supplement containing 1491 mg of DHA and 351 mg of eicosapentaenoic acid (EPA) per day or a placebo supplement containing sunflower oil. Outcome measures included the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS), the National Adult Reading Test (NART), the Coin Rotation Task (CRT), the California Older Adult Stroop Test (COAST), the Trail Making Test A and B (TMT), Digit Span Backwards Test (DSBT), the Geriatric Depression Scale (GDS), the Geriatric Anxiety Inventory (GAI), the 36-Item Short Form Health Survey Questionnaire (SF-36), height, weight and blood pressure as well as red blood cell (RBC) fatty acid profiles. ANCOVAs, t-tests and chi-square tests were used to test for differences between the DHA and placebo groups. The secondary aim of this thesis was to conduct an online cross-sectional survey to investigate how popular fish oil supplements are within a sample population of New Zealanders, to determine why fish oil supplements are used, what dose fish oil users take, and where fish oil users store their supplements. The aim of the survey was to test the hypothesis that fish oil users are more likely to display healthier dietary and lifestyle habits. The final analysis (n = 60) of the trial found no evidence of a treatment effect using the cognitive measures, although it did find a treatment effect on systolic blood pressure (p = .03, ƞ2 = .08), and a treatment interaction for APOE ɛ4 carriers on depression (p = .04, ƞ2= .07) and anxiety (p = .02, ƞ2 = .09) scores in favour of the DHA group. The survey found that in a sample of 334 New Zealanders, 66.8% reported using supplements and 21.9% reported using fish oil supplements. The survey found that respondents who regularly eat oily fish are most likely to take fish oil supplements (p < .01), and that only 26% of fish oil users reported taking a dose that would meet dietary recommendations. Over half of fish oil users reported taking fish oils to ‘improve brain functioning’ and only 6.8% of fish oil users reported storing fish oil supplements in the refrigerator. The evidence from epidemiological studies suggests that fish consumption is protective against cognitive decline and dementia, and yet the evidence from clinical trials investigating the cognitive effects of n-3 PUFAs in older adults has been inconsistent and at times conflicting. It appears that the protective effect of fish may be more than the n-3 PUFA content, and that fish consumption may be part of an overall healthier diet and lifestyle, along with regular physical activity, strong social connections and an emphasis on educational attainment and engagement in cognitive activities.
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    The role of vitamin D and Omega-3 long chain polyunsaturated fatty acids in children with Autism Spectrum Disorder : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Nutritional Science, Massey University, Albany, New Zealand
    (Massey University, 2018) Mazahery, Hajar
    Background: The efficacy of vitamin D and omega-3 long chain polyunsaturated fatty acid (omega-3 LCPUFA), each individually, in Autism Spectrum Disorder (ASD) has been tested in a few trials and the results are inconclusive. Furthermore, several observational studies have observed low vitamin D and omega-3 LCPUFA status in populations with ASD. Children with ASD are susceptible to nutritional issues and poor diet quality due to sensory, behavioural and gastrointestinal issues associated with the condition, though no information regarding these children’s nutritional status is available in New Zealand. Also, no validated nutritional quality assessment tools are available for this population. Aim and Objectives: The overall aim of this study was to investigate the role of vitamin D (VID), omega-3 LCPUFA (OM), or both (VIDOM) in ASD in children through systematically reviewing literature and conducting an intervention trial with these nutrients. The primary objective was to investigate the efficacy of vitamin D, omega-3 LCPUFA, both on core symptoms and sensory issues after correcting major nutritional deficiencies and secondary objectives were to investigate the efficacy of intervention on irritability and hyperactivity, to study dietary adequacy/nutritional status of children with ASD, and to validate a dietary Index of Children’s Eating (DICE) questionnaire against 4-day estimated food record (4DFR). Methods/Design: New Zealand children with ASD (age 2.5-8.0 years) participated in a 12-month randomised, double-blind, placebo-controlled, 2x2 factorial trial. Prior to trial entry, children’s dietary adequacy and nutritional status were assessed by 4DFR, DICE questionnaire (designed based on New Zealand Ministry of Health Food and Nutrition guidelines), and nutritional biomarkers (25(OH)D, red blood cell fatty acids, iron, calcium, albumin, vitamin B12, and folate). Data regarding dietary supplement use and special/exclusion diet, demographics and anthropometrics (height and weight) were also collected. Children then were randomly assigned to one of four treatment groups; daily 2000IU vitamin D3, 722 mg docosahexaenoic acid (DHA), both supplements, or placebo, and behaviours were assessed. Core symptoms were assessed using Social Responsiveness Scale (SRS), sensory issues using Sensory Processing Measure (SPM), problem behaviours including irritability and hyperactivity using Aberrant Behaviour Checklist (ABC). Outcome measures were analysed pre- and post-intervention. Pair-wise mixed effects longitudinal models were used for data analysis. Results: 309 families registered their interest in the study, of whom 190 families were either excluded or not enrolled. The children of remaining families (n=119) were screened for nutritional deficiencies and high serum 25(OH)D concentrations, of whom two children were excluded. Overall, 62% (73/117) of children completed the trial (placebo 16, VID 19, OM 23, VIDOM 15). The mean serum 25(OH)D concentrations (nmol/L) increased in the VID (27±14, P<0.001) and VIDOM (36±17, P<0.001) groups and changed slightly in the OM (1.1±14, P>0.05) and placebo (8.9±23, P>0.05) groups. The median omega-3 index (%) increased in the OM [4.4 (3.3, 5.9), P<0.001] and VIDOM [4.0 (2.0, 6.0), P<0.001] groups and decreased in the VID [-0.2 (-1.0, 0.1)] and placebo [-0.5 (-0.9, -0.1), P>0.05] groups. Compared to placebo, a greater improvement in multiple outcomes in the intervention groups was observed: SRS-social awareness for OM (0.4±2.9 vs. -1.4±2.3, P=0.03) and VIDOM (0.4±2.9 vs. -1.7±3.5, P=0.03); SRS-social communicative functioning for VIDOM (-5.6±10 vs. -16±24, P=0.07); SRS-total for OM (-5.8±12 vs. -17±18, P=0.08); SPM-taste and smell for VIDOM (-0.3±1.7 vs. -2.5±4.3, P=0.06), SPM-balance and motion for OM (-0.1±4.7 vs. -2.6±4.3, P=0.09), ABC-irritability for VID (0.8±6.1 vs. -4.0±4.9 P=0.01) and OM (0.8±6.1 vs. -5.0±5.0, P=0.001); and ABC-hyperactivity for VID (-0.8±5.6 vs. -5.2±6.3, P=0.047). Out of 86 children whose food records were available, approximately 50% (39/86) reported taking dietary supplements and 15% (13/86) were on a special/exclusion diet. A large proportion of children had dietary intake for vitamin D below the Adequate Intake (AI, 96%), protein below the Average Macronutrient Distribution Range (AMDR, 65%), and iodine below the Estimated Average Requirement (EAR, 54%). Dietary intake of fibre (43%) and vitamin E (37%) was also below the AI by at least one third of children. All or most children exceeded the recommendations for sodium (100%), total saturated fat (80%) and sugar (52%). There was a significant and positive correlation (r=0.7; P<0.001) and good agreement (ĸ=0.6) between total scores from DICE (64±16) and 4DFR (58±11). Participants in the highest tertile of DICE had higher intakes of magnesium (P=0.02), vitamin A (P=0.03) and fibre (P=0.06). Conclusions: Vitamin D and omega-3 LCPUFA, each individually or together, improved some behavioural symptoms of ASD. However, large attrition rates and resultant loss of statistical power preclude definitive conclusion and warrant further trials. Also, the baseline assessment of nutrition confirms nutritional issues and poor diet quality in children with ASD. Given the importance of nutrition in growth and development and in the management of ASD, screening of the nutritional status of children with ASD for nutrient adequacy to reduce under- or over-consumption of nutrients is recommended. DICE is a valid tool for the assessment of diet quality in children with ASD living in New Zealand.
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    Identification of biomarkers of colitis to monitor effects of dietary omega-3 polyunsaturated fatty acids in the interleukin-10 gene-deficient mouse model of inflammatory bowel diseases : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Nutritional Science at Massey University, Manawatū, New Zealand
    (Massey University, 2016) Berger, Nadja
    Inflammatory bowel diseases (IBD) are characterised by chronic inflammation of the gastrointestinal tract including the colon (colitis). Increased dietary intake of salmon, which is rich in eicosapentaenoic acid (EPA), was well tolerated by IBD patients, leading to a perceived decrease in symptoms. However, better knowledge of the mechanisms by which EPA-rich diets affect IBD severity, and appropriate biomarkers for assessing these effects, are needed for potential targeted nutritional interventions. This dissertation aimed to determine the temporal effects (early (9 weeks of age) vs. established (12 weeks)) of a diet containing 3.7% EPA, and the dose-dependent effects (15% to 45%) of a salmon diet at 12 weeks of age, on the severity of colitis. Molecular responses in colon and/or liver of the interleukin-10 gene-deficient (Il10-/-) mouse model of IBD and healthy mice were assessed. Caecum digesta, urine and blood were mined to identify biomarkers (microbiota, metabolites and genes) of these responses. The EPA diet reduced the severity of colitis only in 12-week-old Il10-/- mice. This response was associated with changes in gene expression associated with lymphocyte function, eicosanoid signalling and peroxisome proliferator-activated receptor gamma signalling. The blood immune cell gene expression profile did not correlate with reduced colitis in these mice, but the urine metabolite profile was related to changes in colonic tryptophan metabolism. The effects of the salmon diets on colitis were dose-dependent in 12-week-old Il10-/- mice. The intermediate amount of salmon (30%) reduced the severity of colitis and lymphocyte-related gene expression, while enhancing genes in metabolic pathways. Tryptophan metabolism was not affected in these mice, but the urinary metabolite profile correlated with effects on hepatic tocopherol metabolism, as shown by reduced abundance of gamma-carboxyethylhydroxychroman glucoside. The abundances of V. akkermansia, Eubacterium spp., and an unclassified Rikenellaceae were further affected in these mice. This is the first report describing molecular responses in the colon and liver of Il10-/- mice fed a salmon diet associated with reduced colitis. Ultimately these responses could be validated for use in humans, and potentially enable management of IBD with diet.
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    Dietary intakes and food sources of omega-6 and omega-3 polyunsaturated fatty acids in pregnant women living in New Zealand : a thesis presented in partial fulfilment of the requirements for the degree of Masters of Science in Human Nutrition at Massey University, Albany, New Zealand
    (Massey University, 2015) Eickstaedt, Michele
    Background/Aims: Adequate intakes of omega-6 (n-6) and omega-3 (n-3) polyunsaturated fatty acids (PUFAs) are required for fetal growth, brain development and to support a healthy pregnancy. This study aimed to investigate dietary intakes and food sources of n-6 and n-3 PUFAs in a cohort of New Zealand (NZ) pregnant women. Method: Pregnant women (n=596) in their third trimester of pregnancy from throughout NZ completed an online validated FFQ to assess PUFA intakes over the past three months. Individual and combined intakes of the main PUFAs (linoleic acid, LA; alpha linolenic acid, ALA; arachidonic acid, AA; eicosapentaenoic acid, EPA; docosahexaenoic acid, DHA) were compared with dietary recommendations using frequency summary statistics. Results: Estimated median [25th, 75th percentile] intakes were: 11,580 [8,840, 15,760]mg/d LA (recommended 10,000mg/d), 1,300 [790, 2,120]mg/d ALA (recommended 1,000mg/d), 90 [60, 110]mg/d AA (upper limit 800mg/d), 180 [90, 460]mg/d total n-3 LC-PUFA (EPA plus DHA) (recommended 500mg/d), 60 [30, 190]mg/d EPA (recommended 220mg/d, and 110 [50, 250]mg/d DHA (recommended 200mg/d), with 30.9% of participants consuming more than 200mg/d DHA. Participants taking PUFA supplements (19.6%) had median intakes of 370 [210, 530]mg/d DHA, with 79.5% meeting DHA recommendations. Participants taking PUFA supplements were 16.5 times more likely to meet recommendations for DHA compared to participants not taking supplements. For participants not taking PUFA supplements (80.4%), DHA intakes were 90 [50, 160]mg/d and only 19% met the recommendations. Across all women fish and seafood were the main contributors of DHA (84.8%) and EPA (82.1%) intakes, yet only 9.5% and 12.2% of women consumed canned fish or fresh/frozen fish respectively at least twice per week. Over half of participants reported intakes of poultry (63.1%) and beef (60.8%) at least twice per week. Red meats and poultry (36.8%) alongside eggs (23.3%) were important sources of AA intakes. Fats and oils largely contributed to LA (43.2%) and ALA (55.7%) intakes. Conclusion: The majority of pregnant women did not meet the recommended intakes for DHA, which may be in part due to low fish/seafood intakes. Women taking PUFA supplements were more likely to meet these recommendations. These findings highlight the need for nutrition advice on the benefits of consuming n-3 LC-PUFA rich foods such as fish/seafood during pregnancy.
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    Investigating the effects of long chain omega-3 fatty acids on primary school achievement : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Education at Massey University, Albany, Auckland, New Zealand
    (Massey University, 2013) Lee, Kerry Maree
    Background: All parents are keen to support their child to learn and grow. A variety of studies have identified benefits to children's cognitive development with omega-3 (ω-3) PUFA supplementation. The majority of these studies however have involved children with learning or behavioural difficulties and have generally utilised highly specific cognitive tests. Few studies have involved healthy normally-achieving mainstream children and even fewer have used classroom tests to identify academic rather than cognitive changes. Aim: The aim of this study was to investigate whether supplementation with ω-3 PUFA (fish oil) affected the academic achievement of 8-13 year old general classroom children. Whether these children, their parents and teachers could detect changes in learning and behaviour attributed to this supplementation was also investigated. Methods: A double-blind randomised placebo controlled study over a 15 week period was undertaken with 209 children. Randomisation was stratified for age and gender. These were healthy normally-achieving mainstream children who attended the same school. Every school day the active group consumed 900 mg of omega-3 whilst the placebo group consumed 900 mg of vegetable oil. Changes to academic ability was investigated using the Thurstone Word Fluency Tests (testing fluency and spelling), the NZ generated asTTle reading test and maths basic facts tests. The daily consumption of foods enriched in -3 PUFA was assessed using food frequency questionnaires at baseline and recording the child’s intake of these foods every day at school for the duration of the study. Possible changes in behaviour and attitude were investigated using children, parent and teacher questionnaires. Findings: The food frequency questionnaire and intake records identified a low consumption of ω-3 PUFA rich foods. Fish oil treatment did not affect fluency and reading compared to placebo treatment. Significant improvements were identified with fish oil compared to placebo in subgroups of 8-9 year olds for an aspect of spelling and in highly numerate and literate children for division. Parents and teachers did not identify any significant differences between treatment groupings when completing the behaviour questionnaire. Children consuming fish oil reported at 4 and 15 weeks significant improvements related to getting along with the others compared to children in the placebo group. This trend was also reflected in the teacher questionnaires regarding child behaviour. Conclusions: Despite some significant improvements being evident, because of the fact that these were only in subgroups and potentially the result of multiple calculations, the notion that omega-3 can influence academic achievement cannot be accepted. These findings however strongly highlight the need for additional research.
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    Vitamin D and omega-3 fatty acid supplements in children with autism spectrum disorder: a study protocol for a factorial randomised, double-blind, placebo-controlled trial.
    (23/06/2016) Mazahery H; Conlon C; Beck KL; Kruger MC; Stonehouse W; Camargo CA; Meyer BJ; Tsang B; Mugridge O; von Hurst PR
    BACKGROUND: There is strong mechanistic evidence to suggest that vitamin D and omega-3 long chain polyunsaturated fatty acids (n-3 LCPUFAs), specifically docosahexaenoic acid (DHA), have the potential to significantly improve the symptoms of autism spectrum disorder (ASD). However, there are no trials that have measured the effect of both vitamin D and n-3 LCPUFA supplementation on autism severity symptoms. The objective of this 2 × 2 factorial trial is to investigate the effect of vitamin D, n-3 LCPUFAs or a combination of both on core symptoms of ASD. METHODS/DESIGN: Children with ASD living in New Zealand (n = 168 children) will be randomised to one of four treatments daily: vitamin D (2000 IU), n-3 LCPUFAs (722 mg DHA), vitamin D (2000 IU) + n-3 LCPUFAs (722 mg DHA) or placebo for 12 months. All researchers, participants and their caregivers will be blinded until the data analysis is completed, and randomisation of the active/placebo capsules and allocation will be fully concealed from all mentioned parties. The primary outcome measures are the change in social-communicative functioning, sensory processing issues and problem behaviours between baseline and 12 months. A secondary outcome measure is the effect on gastrointestinal symptoms. Baseline data will be used to assess and correct basic nutritional deficiencies prior to treatment allocation. For safety measures, serum 25-hydroxyvitamin D 25(OH)D and calcium will be monitored at baseline, 6 and 12 months, and weekly compliance and gastrointestinal symptom diaries will be completed by caregivers throughout the study period. DISCUSSION: To our knowledge there are no randomised controlled trials assessing the effects of both vitamin D and DHA supplementation on core symptoms of ASD. If it is shown that either vitamin D, DHA or both are effective, the trial would reveal a non-invasive approach to managing ASD symptoms. TRIAL REGISTRATION: Australian New Zealand Clinical Trial Registry, ACTRN12615000144516 . Registered on 16 February 2015.