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    Bioaccessibility and associated concepts: Terminology in the context of in vitro food digestion studies
    (Elsevier Ltd, 2025-09-01) Grundy MM-L; Deglaire A; Le Feunteun S; Reboul E; Moughan PJ; Wilde PJ; McClements DJ; Marze S
    In vitro gastrointestinal models are widely used to study food digestion, in combination with analytical methods to determine the physicochemical and biochemical fate of food compounds. The in vitro bioaccessibility determined with these models is often used as an indicator of the in vivo bioavailability. However, the bioaccessibility concept is not used consistently within the scientific literature, leading to confusion and making it difficult to compare the results from different studies. The aim of this article is to provide standardized definitions of in vitro digestibility and bioaccessibility, detailing the main processes involved, including physical release, solubilization, and biochemical/metabolic reactions. The terminology of complementary cellular, ex vivo, and animal/human in vivo experiments is also given. Application of the in vitro terminology to different nutrients is discussed, including lipids, proteins, carbohydrates, vitamins, and other bioactive compounds. The proposed definitions unify most concepts related to the gastrointestinal fate of ingested food compounds.
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    Effect of age on the standardized ileal amino acid digestibility of soybean meal and canola meal in broilers
    (Elsevier B.V. on behalf of KeAi Communications Co., Ltd., 2023-12-02) Barua M; Abdollahi MR; Zaefarian F; Wester TJ; Girish CK; Chrystal PV; Ravindran V
    Standardized ileal digestibility coefficients (SIDC) of nitrogen (N) and amino acids (AA) in two protein sources (soybean meal [SBM] and canola meal [CM]) were investigated at six broiler ages (d 7, 14, 21, 28, 35, and 42). Two assay diets were formulated to contain either SBM (413 g/kg) or CM (553 g/kg) as the sole dietary AA source. Titanium dioxide (5 g/kg) was added as an indigestible marker. A total of 696 male broilers at 1 d old were allotted to 12 replicate cages per age group. Each assay diet was offered to birds for 4 d prior to the ileal digesta collection on d 7 (14 birds/cage), 14 (12 birds/cage), 21 (10 birds/cage), 28 (8 birds/cage), 35 (8 birds/cage) and 42 (6 birds/cage), respectively. The apparent digestibility coefficients were standardized using age-specific basal endogenous AA flows. In the SBM group, though the SIDC of N tended to be influenced (quadratic; P = 0.075) by age, no linear or quadratic response of age effect was observed on the average SIDC of indispensable (IAA) and total AA (TAA). An age effect (quadratic; P < 0.05) was observed on the average SIDC of dispensable AA (DAA) in SBM with the highest value recorded at d 7, followed by a decrease from d 14 to 28, which increased beyond d 35. The SIDC of some individual AA (Arg, Thr, Trp, Cys, Pro) were affected (P < 0.05 or P < 0.001) in a quadratic manner by age. In the CM, the SIDC of N, average SIDC of IAA, DAA and TAA were influenced (quadratic; P < 0.05 or P < 0.001) by age. The SIDC of N and average SIDC of DAA and TAA were higher from d 7 to 14, declined at d 21, and then increased beyond d 28. The average SIDC of IAA was low between d 7 and 28 and increased thereafter. The SIDC of individual AA were affected (linear or quadratic; P < 0.05 or P < 0.001) by different magnitudes by age. The age influence on the SIDC AA was variable, depending on the protein source and AA. The results demonstrate that age-specific SIDC AA data might need consideration in broiler feed formulations.
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    In vitro digestibility of dietary proteins and in vitro DIAAS analytical workflow based on the INFOGEST static protocol and its validation with in vivo data
    (Elsevier Ltd, 2023-03-15) Sousa R; Recio I; Heimo D; Dubois S; Moughan PJ; Hodgkinson SM; Portmann R; Egger L
    The FAO recommends the digestible indispensable amino acid score (DIAAS) to determine protein quality in foods, preferably tested in vivo. Here, the INFOGESTin vitrodigestion protocol was applied and supplemented with an analytical workflow allowing the assessment of protein digestibility and DIAAS calculation. The protocol was applied to selected samples WPI, zein, collagen, black beans, pigeon peas, All-Bran®, and peanuts. The total protein digestibility, digestibility of individual amino acids (AA), and DIAAS values were established and compared with in vivo data for the same substrates. Total protein digestibility (total Nitrogen, r = 0.7, P < 0.05; primary amines (OPA), r = 0.6, P < 0.02; total AA, r = 0.6, P < 0.02) and digestibility of individual AA (r = 0.6, P < 0.0001) were in good agreement, between in vitro and in vivo, with a mean difference of 1.2 %. In vitro DIAAS was highly correlated with DIAAS obtained from in vivo true ileal digestibility values (r = 0.96, R2 = 0.89, P < 0.0001) with a mean difference of 0.1 %.
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    Protein quality as a complementary functional unit in life cycle assessment (LCA).
    (Springer Nature, 2022-12-28) McAuliffe GA; Takahashi T; Beal T; Huppertz T; Leroy F; Buttriss J; Collins AL; Drewnowski A; McLaren SJ; Ortenzi F; van der Pols JC; van Vliet S; Lee MRF
    GOAL AND THEORETICAL COMMENTARY: A number of recent life cycle assessment (LCA) studies have concluded that animal-sourced foods should be restricted-or even avoided-within the human diet due to their relatively high environmental impacts (particularly those from ruminants) compared with other protein-rich foods (mainly protein-rich plant foods). From a nutritional point of view, however, issues such as broad nutrient bioavailability, amino acid balances, digestibility and even non-protein nutrient density (e.g., micronutrients) need to be accounted for before making such recommendations to the global population. This is especially important given the contribution of animal sourced foods to nutrient adequacy in the global South and vulnerable populations of high-income countries (e.g., children, women of reproductive age and elderly). Often, however, LCAs simplify this reality by using 'protein' as a functional unit in their models and basing their analyses on generic nutritional requirements. Even if a 'nutritional functional unit' (nFU) is utilised, it is unlikely to consider the complexities of amino acid composition and subsequent protein accretion. The discussion herein focuses on nutritional LCA (nLCA), particularly on the usefulness of nFUs such as 'protein,' and whether protein quality should be considered when adopting the nutrient as an (n)FU. Further, a novel and informative case study is provided to demonstrate the strengths and weaknesses of protein-quality adjustment. CASE STUDY METHODS: To complement current discussions, we present an exploratory virtual experiment to determine how Digestible Indispensable Amino Acid Scores (DIAAS) might play a role in nLCA development by correcting for amino acid quality and digestibility. DIAAS is a scoring mechanism which considers the limiting indispensable amino acids (IAAs) within an IAA balance of a given food (or meal) and provides a percentage contribution relative to recommended daily intakes for IAA and subsequent protein anabolism; for clarity, we focus only on single food items (4 × animal-based products and 4 × plant-based products) in the current case exemplar. Further, we take beef as a sensitivity analysis example (which we particularly recommend when considering IAA complementarity at the meal-level) to elucidate how various cuts of the same intermediary product could affect the interpretation of nLCA results of the end-product(s). RECOMMENDATIONS: First, we provide a list of suggestions which are intended to (a) assist with deciding whether protein-quality correction is necessary for a specific research question and (b) acknowledge additional uncertainties by providing mitigating opportunities to avoid misinterpretation (or worse, dis-interpretation) of protein-focused nLCA studies. We conclude that as relevant (primary) data availability from supply chain 'gatekeepers' (e.g., international agri-food distributors and processors) becomes more prevalent, detailed consideration of IAA provision of contrasting protein sources needs to be acknowledged-ideally quantitatively with DIAAS being one example-in nLCA studies utilising protein as a nFU. We also contend that future nLCA studies should discuss the complementarity of amino acid balances at the meal-level, as a minimum, rather than the product level when assessing protein metabolic responses of consumers. Additionally, a broader set of nutrients should ideally be included when evaluating "protein-rich foods" which provide nutrients that extend beyond amino acids, which is of particular importance when exploring dietary-level nLCA.
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    Modifying quinoa protein for enhanced functional properties and digestibility: A review
    (Elsevier B V, 2023-10-05) Cui H; Li S; Roy D; Guo Q; Ye A
    Quinoa (Chenopodium quinoa Willd.) is a pseudocereal plant that originally came from South America. The trend of consuming quinoa is propelled by its well‒balanced amino acid profile compared to that of other plants. In addition, its gluten‒free nature makes quinoa a promising diet option for celiac disease patients. Protein accounts for approximately 17% of the quinoa seed composition and quinoa protein possesses excellent quality. Quinoa protein is mainly composed of 11S globulins (37%) and 2S albumins (35%), both of which are stabilized by disulfide bonds. To date, the alkaline extraction method is the most commonly used method to extract quinoa protein. The functional properties and digestibility of quinoa protein can be improved with the help of various modification methods, and as a result, the application of quinoa protein will be extended. In this review, the extraction method, modification of functional properties and digestibility of quinoa protein are thoroughly discussed, providing insights into the application of quinoa protein in plant‒based foods.
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    Hindgut digestibility in the dog (Canis familiaris) : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Animal Science at Massey University, Palmerston North, New Zealand
    (Massey University, 2006) Singh, Ishwani
    Digestibility trials are important tools used by nutritionists to establish the nutrient requirements of an animal. The most common method used is total faecal collection, which involves the total collection of faeces over a limited period of time. Digestibility trials can also use indigestible markers, such as chromium oxide and titanium oxide, which eliminate the need for a total faecal collection and instead uses sub-sampling methods. The major aim of this thesis was to compare the suitability of chromium oxide (Cr₂0₃) and titanium dioxide (Ti0₂) as indigestible markers in dogs. Due to constraints in the study design (limited space in the facility and therefore a requirement to house dogs in pairs), it needed to be established if Ti0₂ interfered with the chemical analysis of Cr₂0₃ before any animal trials were undertaken. Different concentrations of both markers were added to freeze dried dog faeces. The indigestible markers were then analysed for in the laboratory and recoveries calculated. lt was established that there was no interference of the analysis by either marker. After a pilot study confirmed that coprophagy did not occur in the dogs, the first study (Chapter 2) used 12 entire female Harrier hounds housed in pairs in 6 concrete floor pens. The dogs were fed twice daily with one of 4 treatment diets; a high nutrient diet containing Cr₂0₃ or Ti0₂ or a low nutrient diet containing Cr₂0₃ or Ti0₂. Daily intake was recorded for each dog. Each dog received each of the 4 diets over 4 consecutive evaluation periods of 14 days each. This study showed that 100% recovery of markers was not achieved in the dogs. Recoveries of Cr₂0₃ were 58% and 76% respectively for high and low nutrient diets, and recoveries of Ti0₂ were 80% and 74% respectively for the same two diets. These results suggest that Ti0₂ is currently the best indigestible marker for use in dogs. Study 2 (Chapter 3) used 5 adult female Beagle dogs with surgically prepared ileal cannulas. The ileal cannulation was conducted according to the method of Walker et al. (1994), and the dogs were housed individually in floor pens in temperature controlled rooms. The dogs were fed 5 commercial AAFCO approved diets with free access to water. Dogs were randomised in a replicated 5x 5 Latin-square design with 14 day periods, consisting of a 10 day adaptation period and a 4 day ileal and faecal collection period. lt was found that there were significant differences between ileal and faecal digestibility of dry matter, crude protein, organic matter and carbohydrates of the nutrients that we tested, indicating that there was significant metabolism of nutrients in the hindgut of the dog.
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    Nutritional evaluation of grain legumes for poultry : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Poultry Nutrition at Massey University, Palmerston North, New Zealand
    (Massey University, 2009) Nalle, Catootjie Lusje
    The nutritional value of faba beans (Vicia faba), Australian sweet lupins (Lupinus angustifolius), white lupins (Lupinus albus) and peas (Pisum sativum) grown in New Zealand for broilers were evaluated in terms of their nutritional characteristics, protein quality (protein efficiency ratio), apparent metabolisable energy, apparent ileal digestibility coefficient of amino acids and the effects on bird performance. The effects of dehulling and extrusion cooking on the nutritive value of legumes were also investigated. The first experiment discussed in Chapter 3 evaluated the effect of cultivars on the nutrient profile and protein quality of chickpeas (Cicer arietinum), Australian sweet lupins, peas and soybeans (Glycine max). With the exception of white lupins, cultivars had no effect on the proximate and fibre composition of grain legumes. Starch was the primary carbohydrate component of chickpeas and peas, whilst non-starch polysaccharides were the major carbohydrates in lupins. The legume proteins were deficient in lysine, methionine, cystine and threonine. No differences were found in protein quality between cultivars of the different grain legume species. The lowest weight gain and protein efficiency ratio, in addition to the highest relative pancreatic weight and mortality rate was found in raw soybeans, suggesting that soybeans contained high a concentration of anti-nutritional factors, such as protease inhibitors. Birds fed chickpeas, lupins and peas had a low mortality rate and relative pancreatic weight, confirming that the level of anti-nutrients in these legume seeds was low. The apparent metabolisable energy and apparent ileal digestibility coefficient of amino acids of faba beans, Australian sweet lupins, white lupins and peas were determined in the second experiment (Chapter 4). Cultivar effect on the apparent metabolisable energy values was observed only for faba beans and white lupins. Faba beans, white lupins and peas had comparable apparent metabolisable energy values, but these values were higher than those of Australian sweet lupins, and lower than that of soybean meal. No cultivar differences were found in the apparent ileal digestibility coefficient of amino acids of grain legumes. The apparent ileal digestibility coefficient of amino acids of both lupin species was found to be comparable to that of soybean meal. The effects of feeding diets containing 200 g/kg faba beans, lupins or peas on the performance, digestive tract development and litter quality of broilers were investigated in the third and fourth trials. In the cage trial (Chapter 5), the results showed that the weight gain of birds fed diets containing grain legumes was similar to that of control diet. Feed intake and feed per gain of birds fed diets containing the majority of grain legume cultivars did not differ from those fed the maize-soy diet. Birds fed diets containing faba beans had more dry and friable excreta compared to other treatment diets. The performance of birds fed diets containing 200 g/kg grain legumes during the 35 d grow-out period, in the floor pen trial (Chapter 6), confirmed the results of the cage trial. In this trial, weight gain and feed per gain of birds fed diets without meat meal were superior to those with meat meal. In cage trials, the modification of some segments of digestive tract development was probably due to the dietary NSP. Whilst in floor pen trial, digestive tract development was not influenced by the inclusion of grain legumes. The effect of methodology of determination (direct vs. difference method) on the apparent ileal digestibility coefficient of amino acids of wheat, maize, Australian sweet lupins, peas and soybean meal for broilers was evaluated in the fifth study (Chapter 7). The influence of methodology on apparent ileal digestibility coefficient of amino acids was found to vary amongst the feed ingredients. In general, the apparent ileal digestibility coefficient of amino acids of test ingredients determined by the difference method was higher than those determined by the direct method, suggesting that the use of the direct method may underestimate the apparent ileal digestibility coefficient of amino acids in low and medium protein ingredients. Data reported in Chapter 8 shows that dehulling increased the apparent metabolisable energy values of faba beans and Australian sweet lupins, but it had no beneficial effect on peas. The increase of apparent metabolisable energy values may be attributed to the decrease in non-starch polysaccharides of these legume seeds after dehulling. The removal of hulls increased the amino acid concentrations, but it had no effect on the apparent ileal digestibility coefficient of most amino acids. These results suggest that dehulling of grain legumes would be nutritionally beneficial and, likely to be economical in view of the improved amino acid concentrations and energy values. The final experiment (Chapter 9) demonstrated that extrusion of peas markedly influenced the content of crude protein, non-starch polysaccharides, starch, and trypsin inhibitors. The soluble non-starch polysaccharides and trypsin inhibitor contents of the majority of extruded pea samples were higher than those of raw peas, but insoluble and total non-starch polysaccharides decreased with extrusion. Extrusion had no effect on the apparent ileal protein digestibility and the apparent metabolisable energy of peas, but it increased ileal starch digestibility.
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    Utilization of sweet potato starch, flour and fibre in bread and biscuits : physico-chemical and nutritional characteristics : a thesis submitted in partial fulfilment of the requirements for the degree of Master of Technology in Food Technology, Massey University
    (Massey University, 2008) Mais, Anton
    Sweet-potato contains a limited amount of protein, although rich in dietary fibre content and carbohydrate, so a successful combination with wheat flour for bread and biscuit production would be nutritionally advantageous. In particular, the role of these ingredients in relating to acceptability of breads and biscuit with higher percentage of sweet potato starch, flour in wheat flour. In this study, starch, flour and residue fibre of three sweet-potato varieties (red, orange and white -types) were studied. The 5 -10% combination levels for biscuit-making were found to be acceptable, without affecting the quality of the biscuit (combination of texture and biscuit size). In bread, bread containing 15% red and white replacement starches and orange replacement flour was found to be acceptable level, without affecting the quality of the bread, in an attempt to replace wheat at higher per cent level. The physicochemical study was complemented with a nutritional study to determine beneficial effects of food rich in dietary fibre and starches, in the context of improving diet related problems. RVA results showed sweet-potato ingredients affected differently the pasting temperature, peak viscosity and final viscosity of the normal wheat flour (p<0.05). Fibre inclusion showed large reduction in viscosity and swelling of sweet potato starch. Biscuits and breads containing sweet-potato starch and flour are low in amylose, and digest slowly because of lowly oriented and ‘crystalline’ areas within the granules enable to swell or to ungelatinised starch granules, whereas wheat control biscuit was able to gelatinised starch and exerted a greater effect upon digestibility. There are many other factors that need to be considered when analysing the in vitro starch digestibility such including amylose content, amylopectin structure and presence of fibre and gelatinising. Sweet-potato starch, flour and fibre addition show least effect on bread texture and size and starch, flour and fibre replacement. However, in in vitro starch digestibility test higher values RSS was recorded for starch addition followed by flour addition.
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    Exploring rumen microbe-derived fibre degrading activities for improving feed digestibility
    (Australasian Dairy Science Symposium, 2014) Moon, CD; Gagic, D; Ciric, M; Noel, S; Summers, EL; Li, D; Atua, RML; Perry, R; Sang, C; Zhang, YL; Schofield, LR; Leahy, SC; Altermann, E; Janssen, PH; Arcus, VL; Kelly, WJ; Waghorn, GC; Rakonjac, J; Attwood, GT; Roche, J
    crucial for optimal dairy productivity. Enzymes produced by rumen microbes are primarily responsible for degrading the complex structural polysaccharides that comprise fibre in the plant cell walls of feed materials. Because rumen microbes have evolved with their ruminant hosts over millions of years to perform this task, their enzymes are hypothesised to be optimally suited for activity at the temperature, pH range, and anaerobic environment of the rumen. However, fibre-rich diets are not fully digested, which represents a loss in potential animal productivity. Thus, there is opportunity to improve fibre utilisation through treating feeds with rumen microbe-derived fibrolytic enzymes and associated activities that enhance fibre degradation. This research aims to gain a better understanding of the key rumen microbes involved in fibre degradation and the mechanisms they employ to degrade fibre, by applying cultivation-based and culture-independent genomics approaches to rumen microbial communities of New Zealand dairy cattle. Using this knowledge, we aim to identify new opportunities for improving fibre degradation to enhance dairy productivity. Rumen content samples were taken over the course of a year from a Waikato dairy production herd. Over 1,000 rumen bacterial cultures were obtained from the plant-adherent fraction of the rumen contents. Among these cultures, two, 59 and 103 potentially new families, genera and species of rumen bacteria were identified, respectively. Many of the novel strains are being genome sequenced within the Hungate 1000 rumen microbial reference genome programme, which is providing deeper insights into the range of mechanisms used by the individual strains for fibre degradation. This information has been used to guide the selection of rumen bacterial strains with considerable potential as fibrolytic enzyme producers in vitro, with the intent of developing the strains so that their enzymes may be used as feed pre-treatments for use on farm. Culture-independent metagenomic approaches were also used to explore the activities involved in fibre degradation from the rumen microbial communities. Functional screening has revealed a range of novel enzymes and a novel fibre disrupting activity. Enrichment for the cell-secreted proteins from the community revealed evidence of a diverse range of cellulosomes, which are cell-surface associated multi-enzyme complexes that efficiently degrade plant cell wall polysaccharides. Biochemical and structural characterisation of these proteins has been conducted. In conclusion, cultivation and culture-independent genomic approaches have been applied to New Zealand bovine rumen microbial communities, and have provided considerable new insights into ruminal fibre degradation processes. Novel activities and bacterial species that display desirable activities on fibrous substrates in vitro are now being explored for their potential to improve ruminal fibre degradation, to allow the development of new technologies that will enhance dairy productivity