The a-linolenic acid content of green vegetables commonly available in Australia

Green vegetable consumption has long been considered to have health benefits mainly due to the vitamins, minerals and phytonutrients (such as vitamin C, folate, antioxidants etc) contained in a vegetable-rich diet. Additionally, green vegetables are known to contain a relatively high proportion of omega-3 polyunsaturated fatty acids (PUFAs), primarily in the form of alpha-linolenic acid (18:3n-3). However, there are no data available on the fatty acid composition and concentration of green vegetables commonly consumed in Australia. The present study determined the fatty acid content of 11 green vegetables that are commonly available in Australia. The total fatty acid concentrations of the vegetables under study ranged from 44 mg/100 g wet weight in Chinese cabbage to 372 mg/100 g in watercress. There were three PUFAs in all vegetables analyzed; these were 16:3n-3, 18:2n-6, and 18:3n-3 fatty acids. Sample vegetables contained significant quantities of 16:3n-3 and 18:3n-3, ranging from 23 to 225 mg/100 g. Watercress and mint contained the highest amounts of 16:3n-3 and 18:3n-3, and parsley had the highest amount of 18:2n-6 in both percentage composition and concentration. Mint had the highest concentration of 18:3n-3 with a value of 195 mg/100 g, while watercress contained the highest concentration of 16:3n-3 at 45 mg/100 g. All 11 green vegetables contained a high proportion of PUFAs, ranging from 59 to 72% of total fatty acids. The omega-3 PUFA composition ranged from 40 to 62% of total fatty acids. Monounsaturated fatty acid composition was less than 6% of total fatty acids. The proportion of saturated fatty acids ranged from 21% in watercress and mint to 32% of total fatty acids in Brussels sprouts. No eicosapentaenoic and docosahexaenoic acids were detected in any of the samples. Consumption of green vegetables could contribute to 18:3n-3 PUFA intake, especially for vegetarian populations.

Lipid and fatty acid digestibility of three oil types in the Australian shortfin eel, Anguilla australis

The lipid and fatty acid digestibilities of three semi-purified, isonitrogenous (48.9–50.8% protein) and isocalorific (19.1–20.8 kJ g⁻¹) diets, in which the lipid source was either cod liver oil (CLO), linseed oil (LO) or sunflower oil (SFO), were estimated in the Australian shortfin eel (Anguilla australis) using chromic oxide as an external marker. Apparent percent protein and energy digestibilities of the diets were not significantly (P>0.05) affected by the lipid source, but the lipid digestibility was. The percent apparent lipid digestibility was lowest in the LO diet (90.2±0.6) and highest in the CLO diet (95.6±0.2).

Not all the fatty acids present in any one diet were recovered in the faecal samples. In diets with CLO, only three saturates (out of five), five monoenes and six (out of 11) PUFAs were detected in faecal samples. With all the diets, 20:0 and 22:0, and none of the n−6 HUFA were detected in the faecal samples. The digestibility of all the fatty acids, except 18:3n−3, was lowest in the diet with LO, and significantly so (P>0.05) from the other diets.

In shortfin eel, there was a trend for the digestibility of saturated fatty acids of diets with the animal oil as the lipid source to decrease with increasing chain length, and in diets with vegetable oil to increase initially and then decrease. A somewhat comparable trend was also evident in respect of monoenes.

When the digestibility of different categories of fatty acids is considered, the digestibility of saturates, monoenes, unsaturates, n−6, PUFA, HUFA and total fatty acid digestibilities of LO diet were the lowest, and differed significantly (P<0.05) from those of the CLO and SFO diets, except in the case of n−3 fatty acids.

Variation in the fatty acid composition of the blubber in Cape fur seals (Arctocephalus pusillus pusillus) and the implications for dietary interpretation

Analysis of the fatty acid (FA) composition of blubber is a valuable tool in interpreting the diet of marine mammals. This technique is based on the principle that particular FA present in prey can be incorporated largely untransformed into predator adipose tissue stores, thereby providing biochemical signatures with which to identify prey species. Several studies of phocid seals and cetaceans have documented vertical stratification in the FA composition of blubber such that inferences about diet may vary greatly depending on the layer of the blubber that is analysed. It is not known whether blubber in otariid seals (fur seals and sea lions) also displays vertical stratification in FA composition. Furthermore, it is not known whether the FA composition of blubber is uniform in these species. In the present study, the vertical and regional variation in FA composition of blubber was investigated in seven adult female Cape fur seals (Arctocephalus pusillus pusillus). The proportion of monounsaturated fatty acids (MUFA) was greater in the outer (43.6±1.3%) than inner portion (40.9±1.2%; t20=5.59, P<0.001) whereas the proportions were greater in the inner than outer portions for saturated fatty acids (23.6±0.5% and 21.9±0.6%, respectively, t20 = 5.31, P<0.001) and polyunsaturated fatty acids (PUFA, 35.5±0.7% and 34.5±0.7%, respectively, t20 = 3.81, P < 0.001). There was an inverse relationship between MUFA and PUFA in the blubber, independent of sampling location. In addition, with the exception of the inner portion from non-lactating females, blubber from the mammary area had the highest proportions of 18:1ω9c and total MUFA, followed by blubber from the rump and neck, suggesting that the deposition and mobilisation of blubber lipids may not be uniform around the body in otariid seals. These results support the need for blubber tissue to be sampled from the same site on animals, and to the full depth of the blubber layer, to minimise variation in FA profiles that could occur if different sites and depths were sampled. Such standardisation of sampling will further aid in interpreting diet in otariid seals using the FA Signature Analysis approach.

Effects of dietary oil source on growth and fillet fatty Acid composition of Murray cod, Maccullochella peelii peelii

The Murray cod, an Australian native freshwater fish, supports a relatively small but increasing aquaculture industry in Australia. Presently, there are no dedicated commercial diets available for Murray cod; instead, nutritionally sub-standard feeds formulated for other species are commonly used. The aim of the present investigation was to assess the suitability of two plant based lipid sources, canola oil (CO) and linseed oil (LO), as alternatives to fish oil for juvenile Murray cod. Five iso-nitrogenous, iso-calorific, iso-lipidic semi-purified experimental diets were formulated with 17% lipid originating from 100% cod liver oil (FO), 100% canola oil, 100% linseed oil and 1 : 1 blends of canola and cod liver oil (CFO) and 1 : 1 blends of linseed and cod liver oil (LFO). Each of the diets was fed to apparent satiation twice daily to triplicate groups of 50 Murray cod with initial mean weights of 6.45 ± 1.59 g for 84 days at 22 °C. Final mean weight, specific growth rate and daily feed consumption were significantly higher for the FO and LFO treatments compared to the LO treatment. Feed conversion and protein efficiency ratios were not significantly different amongst treatments. Experimental diets containing vegetable oil and vegetable oil blend(s) had significantly higher concentrations of n-6 fatty acids, predominantly in the form of linoleic acid (LA), while n-3 fatty acids were present in significantly higher concentrations in LO and LFO treatments. The fatty acid composition of Murray cod fillet was reflective of the dietary lipid source. Fillet of fish fed the FO was highest in EPA (20:5n-3), ArA (20:4n-6) and DHA (22:6n-3). Fish fed the CO diet had high concentrations of oleic acid (OlA) (192.2 ± 10.5 mg g lipid− 1), while the fillet of Murray cod fed the LO diet was high in α-linolenic acid (LnA) (107.1 ± 6.7 mg g lipid− 1). The present study suggests that fish oil can be replaced by up to 100% with canola oil and by up to 50% with linseed oil in Murray cod diets with no significant effect on growth.

Growth performance, feed efficiency and fatty acid composition of juvenile Murray cod, Maccullochella peelii peelii, fed graded levels of canola and linseed oil

n two independent experiments, the effects of dietary inclusion of canola and linseed oil were evaluated in juvenile Murray cod (Maccullochella peelii peelii, Mitchell) over a 112-day period. In each experiment, fish received one of five semi-purified diets in which the dietary fish oil was replaced with canola oil (Experiment A) or linseed oil (Experiment B) in graded increments of 25% (0–100%). Murray cod receiving the graded canola and linseed oil diets ranged in final weight from 112.7 ± 7.6 to 73.8 ± 9.9 g and 93.9 ± 3.6 to 74.6 ± 2.2 g, respectively, and exhibited a negative trend in growth as the inclusion level increased. The fatty acid composition of the fillet and liver were modified extensively to reflect the fatty acid composition of the respective diets. Levels of oleic acid (18:1 n-9) and linoleic acid (18:2 n-6) increased with each level of canola oil inclusion while levels of α-linolenic acid (18:3 n-3) increased with each level of linseed oil inclusion. The concentration of n-3 highly unsaturated fatty acids in the fillet and liver decreased as the amount of vegetable oil in the diets increased. It is shown that the replacement of fish oil with vegetable oils in low fish meal diets for Murray cod is possible to a limited extent. Moreover, this study reaffirms the suggestion for the need to conduct ingredient substitution studies for longer periods and where possible to base the conclusions on regression analysis in addition to anova.

Gas chromatography-chemical ionization-mass spectrometric fatty acid analysis of a commercial supercritical carbon dioxide lipid extract from New Zealand green-lipped mussel (Perna canaliculus)

Supercritical fluid extracts of New Zealand green-lipped mussels (NZGLM) have been suggested to have therapeutic properties related to their oil components. The large number of minor FA in NZGLM extract was characterized by a GC-CIMS/MS method that excels at identification of double-bond positions in FAME. The extract contained five major lipid classes: sterol esters, TAG, FFA, sterols, and polar lipids. The total FA content of the lipid extract was 0.664 g/mL. Fifty-three unsaturated FA (UFA) were fully identified, of which 37 were PUFA, and a further 21 UFA were detected for which concentrations were too low for assignment of double-bond positions. There were 17 saturated FA, with 14∶0, 16∶0, and 18∶0 present in the greatest concentration. The 10 n−3 PUFA detected included 20∶5n−3 and 22∶6n−3, the two main n−3 FA; n−3 PUFA at low concentrations were 18∶3, 18∶4, 20∶3, 20∶4, 21∶5, 22∶5, 24∶6, and 28∶8. There were 43 UFA from the n−4, n−5, n−6, n−7, n−8, n−9, n−10, n−11 families, with 16∶2n−4, 16∶1n−5, 18∶1n−5, 18∶2n−6, 20∶4n−6, 16∶1n−7, 20∶1n−7, 16∶1n−9, 18∶1n−9, and 20∶1n−9 being the most abundant. In general, we estimated that FAME concentrations greater than 0.05% (w/w) were sufficient to assign double-bond positions. In total, 91 FA were detected in an extract of the NZGLM, whereas previous studies of fresh flesh from the NZGLM had reported identification of 42 FA. These data demonstrate a remarkable diversity of NZGLM FA.

Omega-3 polyunsaturated fatty acid contents of canned meats avaialbe in Australia

Total lipid content of 20 species of canned meats available in Australia ranged from 2% in chicken (Hormel, USA) to 41% in stewed pork (Ma Ling, China). Total n-3 polyunsaturated fatty acids (PUFA) ranged from 30 in canned chicken (Hormel) to 659 mg/100 g in chicken hot dog (Tulip, Denmark). The 18:2n-6 was the predominant PUFA, ranging from 187 in corned beef (Hamper, Australia) to 2832 mg/100 g in chicken luncheon meat (Tulip). Other main PUFA, in order of concentration, were 18:3n-3, ranging from 14 in canned chicken (Hormel) to 590 mg/100 g in chicken hot dog (Tulip); conjugated 18:2n-6 (CLA) from 1 in chicken (Hormel) to 135 mg/100 g in corned mutton (Colonial, Australia); 20:4n- 6 from 11 in camp pie (Tom Piper, Australia) to 73 mg/100 g in spiced ham (Hormel); and 22:5n-3 from 5 in chicken (Hormel) and chicken luncheon (Almaraai, Jordan) to 45 mg/100 g in stewed pork (Ma Ling). Total saturated fatty acids (SFA) ranged from 598 to 14 660 mg/100 g, with 16:0 predominant followed by 18:0. Total monounsaturated fatty acid concentration ranged between 813 to 20 218 mg/100 g with 18:1 the major fatty acid. Trans 18:1 ranged from 10 to 698 mg/100 g. The canned meats contained 20 and 22-carbon long chain n-3 PUFA at levels comparable with or greater than those in fresh lean meat.

Fatty acid composition of habitual omnivore and vegetarian diets

High-fat diets are implicated in the onset of cardiovascular disease (CVD), cancer, and obesity. Large intakes of saturated and trans FA, together with low levels of PUFA, particularly long-chain (LC) omega-3 (n−3) PUFA, appear to have the greatest impact on the development of CVD. A high n−6∶n−3 PUFA ratio is also considered a marker of elevated risk of CVD, though little accurate data on dietary intake is available. A new Australian food  composition database that reports FA in foods to two decimal places was used to assess intakes of FA in four habitual dietary groups. Analysis using the database found correlations between the dietary intakes of LC n−3 PUFA and the plasma phospholipid LC n−3 PUFA concentrations of omnivore and vegetarian subjects. High meat-eaters (HME), who consumed large  amounts of food generally, had significantly higher LC n−3 PUFA intakes (0.29 g/d) than moderate meat-eaters (MME) (0.14 g/d), whose intakes in turn were significantly higher than those of ovolacto-vegetarians or vegans (both 0.01 g/d). The saturated FA intake of MME subjects (typical of adult male Australians) was not different from ovolacto-vegetarian intakes, whereas n−6∶n−3 intake ratios in vegetarians were significantly higher than in omnivores. Thus, accurate dietary and plasma FA analyses suggest that regular moderate consumption of meat and fish maintains a plasma FA profile possibly more conducive to good health.

Interacting amino acid preferences of 3D pattern pairs at the binding sites of transient and obligate protein complexes

To assess the physico-chemical characteristics of protein-protein interactions, protein sequences and overall structural folds have been analyzed previously. To highlight this, discovery and examination of amino acid patterns at the binding sites defined by structural proximity in 3-dimensional (3D) space are essential. In this paper, we investigate the interacting preferences of 3D pattern pairs discovered separately in transient and obligate protein complexes. These 3D pattern pairs are not necessarily sequence-consecutive, but each residue in two groups of amino acids from two proteins in a complex is within certain °A threshold to most residues in the other group. We develop an algorithm called AA-pairs by which every pair of interacting proteins is represented as a bipartite graph, and it discovers all maximal quasi-bicliques from every bipartite graph to form our 3D pattern pairs. From 112 and 2533 highly conserved 3D pattern pairs discovered in the transient and obligate complexes respectively, we observe that Ala and Leu is the highest occuring amino acid in interacting 3D patterns of transient (20.91%) and obligate (33.82%) complexes respectively. From the study on the dipeptide composition on each side of interacting 3D pattern pairs, dipeptides Ala-Ala and Ala-Leu are popular in 3D patterns of both transient and obligate complexes. The interactions between amino acids with large hydrophobicity difference are present more in the transient than in the obligate complexes. On contrary, in obligate complexes, interactions between hydrophobic residues account for the top 5 most occuring amino acid pairings.

Abscisic acid regulation of plant defence responses during pathogen attack

The plant hormone, abscisic acid (ABA), has previously been shown to have an impact on the resistance or susceptibility of plants to pathogens. In this thesis, it was shown that ABA had a regulatory effect on an extensive array of plant defence responses in three different plant and pathogen interaction combinations as well as following the application of an abiotic elicitor. In unique studies using ABA deficient mutants of Arabidopsis, exogenous ABA addition or ABA biosynthesis inhibitor application and simulated drought stress, ABA was shown to have a profound effect on the outcome of interactions between plants and pathogens of differing lifestyles and from different kingdoms. The systems used included a model plant and an important agricultural species: Arabidopsis thaliana (Arabidopsis) and Peronospora parasitica (a biotrophic Oomycete pathogen), Arabidopsis and Pseudomonas syringae pathovar tomato (a biotrophic bacterial pathogen) and an unrelated plant species, soybean (Glycine max) and Phytophthora sojae (a hemibiotrophic Oomycete pathogen), Generally, a higher than basal endogenous ABA concentration within plant tissues at the time of avirulent pathogen inoculation, caused an interaction shift towards what phenotypically resembled susceptibility. Conversely, a lower than basal endogenous ABA concentration in plants inoculated with a virulent pathogen caused a shift towards resistance. An extensive suppressive effect of ABA on defence responses was revealed by a range of techniques that included histochemical, biochemical and molecular approaches. A universal effect of ABA on suppression or induction of the phenylpropanoid pathway via regulation of the key entry point gene, phenylalanine ammonia-lyase (PAL), when stimulated by biotic or abiotic elicitors was shown. ABA also influenced a wide variety of other defence-related components such as: the development of a hypersensitive response (HR), the accumulation of the reactive oxyden species, hydrogen peroxide and the cell wall strengthening compounds lignin and callose, accumulation of SA and the phytoalexin, glyceollin and the transcription of the SA-dependent pathogenesis- related gene (PR-1). The near genome-wide microarray gene expression analysis of an ABA induced susceptible interaction also revealed an yet unprecedented insight into the great diversity of defence responses that were influenced by ABA that included: disease resistance like proteins, antimicrobial proteins as well as phenylpropanoid and tryptophan pathway enzymes. Subtle differences were found in the number and type of defence responses that were regulated by ABA in each type of plant and pathogen interaction that was studied. This thesis has clearly identified in plant/pathogen interactions previously unknown and important roles for ABA in the regulation of many defence responses.

Hypertension induced by ω-3 polyunsaturated fatty acid deficiency is alleviated by α-linolenic acid regardless of dietary source

Ω-3 polyunsaturated fatty acid deficiency, particularly during the prenatal period, can cause hypertension in later life. This study examined the effect of different sources of α-linolenic acid (canola oil or flaxseed oil) in the prevention of hypertension and other metabolic symptoms induced by an ω-3 fatty acid-deficient diet. Dams were provided one of three experimental diets from 1 week before mating. Diets were either deficient (10% safflower oil-DEF) or sufficient (7% safflower oil+3% flaxseed oil-SUF-F; or 10% canola oil-SUF-C) in ω-3 fatty acids. The male offspring were continued on the maternal diet from weaning for the duration of the study. Body weight, ingestive behaviors, blood pressure, body composition, metabolic rate, plasma leptin and brain fatty acids were all assessed. The DEF animals were hypertensive at 24 weeks of age compared with SUF-F or SUF-C animals; this was not evident at 12 weeks. These results suggest that different sources of ALA are effective in preventing hypertension related to ω-3 fatty acid deficiency. However, there were other marked differences between the DEF and, in particular, the SUF-C phenotype including lowered body weight, adiposity, leptin and food intake in SUF-C animals. SUF-F animals also had lower, but less marked reductions in adiposity and leptin compared with DEF animals. The differences observed between DEF, SUF-F and SUF-C phenotypes indicate that body fat and leptin may be involved in ω-3 fatty acid deficiency hypertension.

Estimating the impact of mandatory folic acid fortification on the folic acid intake of Australian women of childbearing age

Objective: The primary aim of this study was to estimate the impact of mandatory folic acid (FA) fortification of bread-making flour on the FA intake of Australian women of childbearing age (16-44 years). The secondary objective was to investigate the relationship between estimated FA intake and socio-economic status (SES) and age.

Method: Dietary modelling was used to estimate FA intake under four mandatory fortification scenarios – no supplement use, supplement use unrelated to FA intake, supplement use only among the highest consumers of bread, and increased supplement use. Data were obtained from the 1995 National Nutrition Survey for food intake patterns, the 2007 Victorian Population Health Survey for FA supplement use, and a marketplace survey.

Results: It is estimated that the National Health and Medical Research Council (NHMRC) recommendation for an additional 400 mg/day FA will be achieved by a minimum of 3.9, 25.4, 21.7 and 30% of the target population under scenarios 1-4, respectively. The FA upper level of intake is exceeded by a maximum of 0.1, 1.7, 6.1 and 4.1% of the target population for scenarios 1-4, respectively.

Conclusions: Mandatory FA fortification is not sufficient for the NHMRC recommendations for minimum and maximum intakes to be met by all of the target population under a number of plausible behaviour scenarios.

Implications: Targeted nutrition education campaigns are needed for SES and age sub-groups and research of this nature should be extended to other population groups. Monitoring and evaluation of this policy will be important to ensure appropriate FA intake.

Effects of Dietary Vitamin B6 Supplementation on Fillet Fatty Acid Composition and Fatty Acid Metabolism of Rainbow Trout Fed Vegetable Oil Based Diets

Fish oil replacement in aquaculture feeds results in major modifications to the fatty acid makeup of cultured fish. Therefore, in vivo fatty acid biosynthesis has been a topic of considerable research interest. Evidence suggests that pyridoxine (vitamin B₆) plays a role in fatty acid metabolism, and in particular, the biosynthesis of LC-PUFA has been demonstrated in mammals. However, there is little information on the effects of dietary pyridoxine availability in fish fed diets lacking LC-PUFA. This study demonstrates a relationship between dietary pyridoxine supplementation and fatty acid metabolism in rainbow trout. In particular, the dietary pyridoxine level was shown to modulate and positively stimulate the activity of the fatty acid elongase and Δ-6 and Δ-5 desaturase enzymes, deduced by the whole-body fatty acid balance method. This activity was insufficient to compensate for a diet lacking in LC-PUFA but does highlight potential strategies to maximize this activity in cultured fish, especially when fish oil is replaced with vegetable oils.