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Two experiments were conducted with spray dried animal plasma (SDAP) in diets for piglets from 0 to 14 days post-weaning. Experiment 1 was conducted to determine a possible interaction between diet composition and SDAP in diets without anti-microbial growth promoter (AMGP). Experiment 2 was conducted to determine a possible interaction between SDAP and AMGP. Both experiments comprised four treatments in a 2×2 factorial arrangement. The respective factors were diet composition (simple vs. complex with more animal protein) and SDAP (0% vs. 4%) in experiment 1 and AMGP (0 vs. 40 ppm avilamycine) and SDAP (0% vs. 4%) in experiment 2. During the first week post-weaning SDAP improved feed intake by 14% in experiment 2 and average daily gain (ADG) and feed conversion ratio (FCR) by approximately 20% and 10%, respectively, in both experiments (P

Complexation of dietary phytate with cations is a major cause of reduced bioavailability of Zn and possibly Cu in pig diets. We conducted 2 studies with 2 treatments in young growing pigs (8 to 40 kg) to estimate potential contributions of phytase to availability and supply of Zn and Cu, respectively. Each treatment comprised 10 pens with 8 pigs each as experimental units. In Exp. 1, 500 phytase units (FTU)/kg of microbial phytase (Natuphos 5000G; BASF) was added to a diet containing 15 mg Zn from ZnSO(4) and 160 mg/kg Cu from CuSO(4) in addition to Cu and Zn from feed ingredients. In Exp. 2, 500 FTU/kg was added to a diet containing 45 mg Zn from ZnSO(4) without added CuSO(4). Feces were collected to determine nutrient digestibility, blood was collected, and pigs were killed to determine Cu and Zn in the liver. In both experiments, phytase inclusion increased (P < 0.001) Zn digestibility by on average 10% units, serum Zn level (P < 0.001) by 0.4 mg/L, and liver Zn content (P < 0.001) by 129 mg/kg DM. In Exp. 1 phytase increased (P = 0.03) Cu digestibility by 6% units but reduced (P = 0.04) liver Cu content by 35 mg/kg DM. In Exp. 2 phytase reduced (P < 0.001) Cu digestibility by 16% units without affecting liver Cu content. Results indicate that the effect of phytase on Cu availability depends on dietary Cu and Zn content and the response variable studied. In conclusion, the consistent effects of phytase on indices of Zn status allow a reduction of Zn inclusion in phytase-supplemented diets.

The livestock sector is in urgent need for more sustainable feed sources, because of the increased demand for animal-source food and the already high environmental costs associated with it. Recent developments indicate environmental benefits of rearing insects for livestock feed, suggesting that insect-based feed might become an important alternative feed source in the coming years. So far, however, this potential environmental benefit of waste-fed insects is unknown. This study, therefore, explores the environmental impact of using larvae of the common housefly grown on poultry manure and food waste as livestock feed. Data were provided by a laboratory plant in the Netherlands aiming to design an industrial plant for rearing housefly larvae. Production of 1 ton dry matter of larvae meal directly resulted in a global warming potential of 770 kg CO2 equivalents, an energy use of 9329 MJ and a land use of 32 m2, caused by use of water, electricity, and feed for flies, eggs and larvae. Production of larvae meal, however, also has indirect environmental consequences. Food waste, for example, was originally used for production of bio-energy. Accounting for these indirect consequences implies, e.g., including the environmental impact of production of energy needed to replace the original bio-energy function of food waste. Assuming, furthermore, that 1 ton of larvae meal replaced 0.5 ton of fishmeal and 0.5 ton of soybean meal, the production of 1 ton larvae meal reduced land use (1713 m2), but increased energy use (21,342 MJ) and consequently global warming potential (1959 kg CO2-eq). Results of this study will enhance a transparent societal and political debate about future options and limitations of larvae meal as livestock feed. Results of the indirect environmental impact, however, are situation specific, e.g. in this study food waste was used for anaerobic digestion. In case food waste would have been used for, e.g., composting, the energy use and related emission of greenhouse gases might decrease. Furthermore, the industrial process to acquire housefly larvae meal is still advancing, which also offers potential to reduce energy use and related emissions. Eventually, land scarcity will increase further, whereas opportunities exist to reduce energy use by, e.g., technical innovations or an increased use of solar or wind energy. Larvae meal production, therefore, has potential to reduce the environmental impact of the livestock sector.

Background Maternal diet during pregnancy can modulate skeletal muscle development of the offspring. Previous studies in pigs have indicated that a fat supplemented diet during pregnancy can improve piglet outcome, however, this is in contrast to human studies suggesting adverse effects of saturated fats during pregnancy. This study aimed to investigate the impact of a fat supplemented (palm oil) “high fat” diet on skeletal muscle development in a porcine model. Histological and metabolic features of the biceps femoris muscle obtained from 7-day-old piglets born to sows assigned to either a commercial (C, n = 7) or to an isocaloric fat supplementation diet (“high fat” HF, n = 7) during pregnancy were assessed. Results Offspring exposed to a maternal HF diet demonstrated enhanced muscular development, reflected by an increase in fractional growth rate, rise in myofibre cross-sectional area, increased storage of glycogen and reduction in lipid staining of myofibres. Although both groups had similar intramuscular protein and triglyceride concentrations, the offspring born to HF mothers had a higher proportion of arachidonic acid (C20:4n6) and a reduction in α-linolenic acid (C18:3n3) compared to C group offspring. The HF group muscle also exhibited a higher ratio of C20:3n6 to C20:4n6 and total n-6 to n-3 in conjunction with up-regulation of genes associated with free fatty acid uptake and biogenesis. Conclusion In conclusion, a HF gestational diet accelerates the maturation of offspring biceps femoris muscle, reflected in increased glycolytic metabolism and fibre cross sectional area, differences accompanied with a potential resetting of myofibre nutrient uptake.

Organic acids in pig diets in some cases enhance growth performance, regulate microbiota in the gastrointestinal tract (GIT), and reduce the occurrence of post-weaning diarrhoea. Organic acids may also increase nutrient digestibility by reducing gastric pH, increasing pepsin activity and reducing stomach passage rate. The objective of this study was to determine the effect of two blends of organic acids on apparent faecal digestibility in weaned piglets. Twenty-four piglets, weaned at 26 days of age, were used in this experiment, comprising three treatments: a control diet (C); C with inclusion of 5 kg/ton of a blend of formic acid and essential oils; C with 5 kg/ton of a blend of formic acid and other organic acids including propionic, lactic, citric and sorbic acid. Piglets were individually housed and fed the experimental diets from days 6 to 30 post-weaning. The combination of formic acid and other organic acids significantly increased faecal digestibility of crude fibre compared to the C (25.7% vs. 22.7%; P

Piglet neonatal mortality rates are high (~20%), so nutritional strategies to reduce this are highly desirable. Maternal fat substitution (FS) may promote the preweaning survival of piglets by improving their energy status. Therefore, the aim of the present study was to investigate the effects of FS throughout pregnancy on offspring viability, together with the gene expression of stress-related markers in the liver. Sixteen pregnant sows were randomly allocated to one of two isocaloric diets, control (C) or FS in the form of palm oil, fed from 0 to 110 days gestation. Glucose tolerance was examined on Day 108. Median and low birthweight offspring were allocated to tissue sampling at either 7 days or 6 months postnatal age. In response to a glucose tolerance test, FS sows exhibited a raised glucose area under the curve with no change in basal glucose. Average piglet mortality (up to Day 28) was increased fourfold in the FS group, with surviving median-sized piglets exhibiting significantly lower fatty acid binding protein 1 (FABP1) expression at 7 days. There were no effects on the abundance of any other stress- or metabolic-related genes examined. Thus, this study demonstrates that maternal FS throughout gestation causes maternal glucose intolerance that may be linked to the observed increase in piglet mortality. However, the surviving offspring do not exhibit any detectable differences in postnatal growth or hepatic gene profile in later life.

The present study evaluated the dose response of Buttiauxella phytase on apparent total tract digestibility (ATTD) of DM, Ca, and P in weaned pigs at 2 locations. Experimental diets fed to weaned pigs were a positive control (PC), a negative control (NC), and NC supplemented with increasing levels of Buttiauxella phytase. In Trial A, ATTD of P was 57.2% for PC, 32.5% for NC, and 59.4, 62.0, 63.8, 66.0, and 67.3% for 250, 500, 750, 1000, and 2000 phytase units (FTU) added to NC, respectively. In Trial B, ATTD of P was 45.2% for PC, 28.4% for NC, and 58.7, 64.1, 67.9, and 70.9% for 250, 500, 1000, and 2000 FTU added to NC, respectively. In both studies, the reduction in P in the NC diets reduced (P < 0.01) ATTD of P when compared with the PC diets. Phytase supplementation linearly and quadratically increased (P < 0.01) ATTD of P at all inclusion levels to the NC diet. In conclusion, the average digestible P increase from Buttiauxella phytase (vs. the NC diet) was 1.3, 1.5, 1.6, and 1.7 g digestible P/kg feed for 250, 500, 1000, and 2000 FTU/kg, respectively.

Feed ingredients, e.g. oil seed by-products, generally have undergone several processing steps before inclusion in animal diets. Processing of feed ingredients and diets can result in conformational changes of protein structure, the formation of Maillard reaction products and cross-links between amino acids (Bender, 1972). These changes affect protein quality and the latter two reactions especially reduce the amount of the essential amino acid lysine (Mauron, 1990). The effects on protein quality may result in an impaired pig performance presumably related to a decrease in protein, amino acid and lysine digestibility as found for processed diets (Gonzalez-Vega et al., 2011). The aims of this research were to determine the effects of processing of two oil seed by-products, i.e. soybean meal (SBM) and rapeseed meal (RSM), on apparent ileal crude protein, amino acid and lysine digestibility and performance in growing pigs and to derive criteria to evaluate protein quality in heat treated ingredients. First results of ileal crude protein digestibility are presented here.

The consequences of sub-optimal nutrition through alterations in the macronutrient content of the maternal diet will not simply be reflected in altered neonatal body composition and increased mortality, but are likely to continue into adulthood and confer greater risk of metabolic disease. One mechanism linking manipulations of the maternal environment to an increased risk of later disease is enhanced fetal exposure to glucocorticoids (GC). Tissue sensitivity to cortisol is regulated, in part, by the GC receptor and 11β-hydroxysteroid dehydrogenase (11β-HSD) types 1 and 2. Several studies have shown the effects of maternal undernutrition, particularly low-protein diets, on the programming of GC action in the offspring; however, dietary excess is far more characteristic of the diets consumed by contemporary pregnant women. This study investigated the programming effects of moderate protein supplementation in pigs throughout pregnancy. We have demonstrated an up-regulation of genes involved in GC sensitivity, such as GC receptor and 11β-HSD, in the liver, but have yet to detect any other significant changes in these piglets, with no differences observed in body weight or composition. This increase in GC sensitivity was similar to the programming effects observed following maternal protein restriction or global undernutrition during pregnancy.

Three experiments were conducted to evaluate the effects of temperature ( T ), pH, time of incubation and pepsin on the stability of intrinsic phytase of different cereals. Experiment one was conducted as a 3×4×3×3 factorial arrangement with three feed ingredients (rye, wheat and barley), four T (20, 38, 55 and 80 °C), three pH levels (3.0, 5.5 and 8.0) and three incubation times (30, 60 and 120 min), with two replicates per treatment. Stability was calculated as the ratio of the residual phytase activity of a treated sample to the non-treated (reference sample) times 100. Phytase activity of the reference samples of rye, wheat and barley was 3.14, 1.77 and 0.66 FTU/g, respectively. Overall, rye showed the highest and barley the lowest stability (0.58 vs. 0.47, P T to 0.07 at 80 °C across ingredients, pH levels and time of incubation (P vs. 0.34 across ingredients, T and time of incubation, P 0.90). Wheat and rye phytases were stable at pH 3.0 and 5.5 (20 and 38 °C). In experiment two, the effect of shorter period of time (10 and 30 min) was assessed on the stability of cereal phytases. Barley phytase showed the highest residual phytase activity among cereals at 65 °C. In experiment three, stability of intrinsic phytase of these feed ingredients was determined after incubation in different concentrations of pepsin (0, 5 and 10 mg/mL) at pH 2.0. Intrinsic phytases of wheat and rye were resistant to pepsin, but barley phytase was susceptible to pepsin and its stability decreased to 0.57 after pre-incubation for 60 min in 5 mg pepsin/mL. This knowledge regarding the effect of T , incubation time and pH on stability of cereal intrinsic phytases may contribute to optimize available P contents of diets, thereby reducing phosphorus excretion of monogastrics.

Increasing the dietary amount of fermentable carbohydrates (FC) may counteract the negative effects of protein fermentation in newly weaned piglets. To study this hypothesis, 272 newly weaned piglets were allotted to 4 dietary treatments in a 2 × 2 factorial arrangement with low and high FC (7.5 and 13.5%) and crude protein (CP, 15 and 22%) content as respective factors. Intestinal histology, enzyme activity, microbiota and fermentation products were determined in 8 pigs per treatment 7 days post-weaning. In the 4 wk experimental period, interactions between dietary CP and FC content were found for feed intake ( P = 0.022), daily gain ( P = 0.001), and gain:feed ( P = 0.033). The high-FC content reduced daily gain by 50 g/d in the low-CP diet, whereas the FC content did not affect growth performance in the high-CP diet. Over the 4 wk experimental period, daily gain (350 g/d) and feed intake (519 g/d) were highest for piglets on the low-CP low-FC diet. The high-FC content resulted in an increase in number of lactobacilli ( P = 0.047) and a decrease of total coliforms ( P = 0.06) in the small intestine. It increased the lactic acid content ( P = 0.08) and reduced the ammonia content ( P = 0.04) in the small intestine and increased the VFA content in the colon ( P = 0.009). The reduction in CP content reduced ammonia concentration in the small intestine ( P = 0.003). We concluded that dietary FC influenced microbial population and fermentation products in the gut. However, this was not reflected in an increased growth performance.

In this article the disturbing steps in the process leading to the Post Weaning Syndrome (PWS) in piglets and its prevention is discussed from a nutritional point of view. The proposed sequence of disturbing steps in the gut is the following: when weaning occurs at three or four weeks of age, gut development to digest a 'vegetable' diet is immature and the normal gut morphology is disturbed by stress, infection, and a low feed intake. The impaired absorption and digestion increases the amount of undigested substrate and may cause explosive growth of bacteria. The colonization resistance is disturbed and for example Escherichia coli can proliferate. Practical measures regarding feeding strategy and feed composition to counteract these disturbing steps in this process leading towards PWS are discussed., Une approche nutritionnelle pour la prévention du syndrome de post-sevrage chez le porcelet. Le syndrome du post-sevrage et sa prévention, chez le porcelet, sont discutés d'un point de vue nutritionnel dans cet article. Les troubles du tube digestif après le sevrage conduisant à ce syndrome sont les suivants : avec un sevrage à 3-4 semaines d'âge, le tube digestif est immature pour digérer les aliments végétaux, la morphologie intestinale est perturbée par le stress, les infections et le faible niveau d'ingestion alimentaire. La digestion et l'absorption des nutriments sont altérées, ce qui augmente la quantité de substrat indigestible et pourrait causer un développement bactérien explosif. La résistance à la colonisation est perturbée et les bactéries pathogènes, comme certaines souches d'Escherichia coli, peuvent proliférer. Des mesures pratiques concernant les stratégies alimentaires et la composition des aliments pour contrecarrer ces perturbations sont discutées.

In 36 growing pigs (30 to 60 kg), N balance and amino acid (AA) composition of weight gain were measured to evaluate the interactive effect of the ratio between N from essential amino acids (EAA(N)) to nonessential amino acids (NEAA(N)) and total N level (T(N)) in the diet on N retention and utilization of N, EAA(N), NEAA(N), and AA. Nine diets composed from ordinary feedstuffs and supplemented with crystalline AA were used (three EAA(N):NEAA(N) ratios of 38:62, 50:50, and 62:38 at three T(N) levels of 18.8, 22.9, and 30.0 g/kg). Pigs were fed restrictedly, at a level of 2.8 x energy for maintenance. In all diets, EAA (including arginine) supply was according to or slightly above the recommended ratios to lysine. Measurements were done in four blocks of nine pigs each. In a concomitant slaughter experiment, the AA composition of deposited body protein was determined to estimate AA utilization. The effects of T(N) and EAA(N):NEAA(N) and their interaction for N retention and utilization were significant. Nitrogen retention increased with higher T(N) in the diet. Increasing EAA(N):NEAA(N) from 38:62 to 50:50 improved N retention only at the two lower T(N) levels. Increasing EAA(N): NEAA(N) above 50:50 failed to improve N retention significantly at any of the three T(N) levels. Lowering T(N) improved the utilization of total and digested N and of EAA(N) and NEAA(N). The increase in EAA(N): NEAA(N) consistently resulted in a lower utilization of EAA(N), but this was compensated by a higher utilization of NEAA(N). The utilization of T(N) was improved by increasing EAA(N):NEAA(N) from 38:62 to 50:50 at the two lower T(N) levels and was relatively unaffected by EAA(N):NEAA(N) at the highest T(N). However, a lower utilization of N was observed at a ratio of 62:38 at a T(N) level of 22.9 g/kg. The effects were similar for utilization of individual EAA and NEAA. Utilization of alanine, aspartic acid, and glycine was close to or >100% at the highest EAA(N):NEAA(N), which was expected because all of these AA are synthesized in pigs. Also, the utilization of arginine was >100% in most of the treatments, which confirms the semiessential character of this AA for maintenance. We concluded that the required ratio of EAA(N):NEAA(N) for optimal N retention and utilization is approximately 50:50. The EAA(N):NEAA(N) is more important at lower dietary protein levels. This study indicates that EAA(N): NEAA(N) can be increased up to 70:30 without lowering the utilization of N. Thus, deaminated EAA(N) was efficiently utilized for the synthesis of NEAA(N).

Ninety-five female pigs from 20 to 45 kg body weight were used to elucidate the effects of energy and protein intake on the amino acid composition of the protein in the carcass, organs and empty body of growing pigs. In a 2 x 15 factorial arrangement, protein intake ranged from 127 to 350 g/d in 15 graduated steps; and the digestible energy allowances were 15.8 and 18.8 MJ/d. Whole-body amino acid contents (g/16 g nitrogen) were (means +/- SEM) lysine 6.64 +/- 0.028, methionine 2.11 +/- 0.012; threonine 3.62 +/- 0.016 and total essential amino acids 42.8 +/- 0.16. The organ fraction contained 14.8 and 15.8% (SEM 0.13) of whole-body protein at the low and high energy levels, respectively. The concentrations of essential amino acids were 41.8 +/- 0.19 and 48.4 +/- 0.13 g/16 g nitrogen in the carcass and organs, respectively. Concentrations of a number of amino acids (in carcass, organ and whole-body protein and in protein deposited between 20 and 45 kg, were affected by protein and/or energy intake. The amino acid pattern of the newly deposited protein was slightly different from that of the empty body protein. The changes in amino acid contents were presumably the result of effects of protein and energy intake on the proportions of muscle and non-muscle carcass tissues and on relative weights of blood and viscera. Consequences of these changes for the amino acid requirements are discussed.

Production of food has re-emerged at the top of the global political agenda, driven by two contemporary challenges: the challenge to produce enough nutritious food to feed a growing and more prosperous human population, and the challenge to produce this food in an environmentally sustainable way. Current levels of production of especially animal-source food (ASF), pose severe pressure on the environment via their emissions to air, water, and soil; and their use of scarce resources, such as land, water, and fossil energy. The livestock sector, for example, is responsible for about 15% of the global anthropogenic emissions of greenhouse gases and uses about 70% of global agricultural land. Many proposed mitigation strategies to feed the world sustainably, therefore, focus primarily on reducing the environmental impact of the livestock sector, so-called production-side strategies. Other strategies focus on changing consumption patterns by reducing consumption of ASF, or on shifting from ASF with a higher environmental impact (e.g. beef) to ASF with a lower environmental impact (e.g. pork or chicken), so called consumption-side strategies. Most of the environmental impact of production of ASF is related to production of feed. One production-side strategy to reduce the environmental impact is the use of products that humans cannot or do not want to eat, such as co-products, food-waste, and biomass from marginal lands for livestock feed (referred to as ‘leftover streams’ in this thesis). This strategy is effective, because feeding leftover streams to livestock transforms an inedible food stream into high-quality food products, such as meat, milk, and eggs. Two production-side strategies that use leftover streams as livestock feed were explored in this thesis: replacing soybean meal (SBM) in diets of growing pigs with either rapeseed meal (RSM) or with waste-fed larvae meal. Replacing SBM with RSM in growing-pig diets was assessed because RSM became increasingly available following an increase in bio-energy production in the EU. In this strategy, therefore, the RSM content in pig diets increased at the expense of SBM. SBM is an ingredient associated with a high environmental impact. It was expected, therefore, that replacing SBM with RSM in pig diets would lead to a decrease in the environmental impact of pork production. Replacing SBM with waste-fed larvae meal was assessed because recent developments show the environmental benefits of rearing insects as livestock feed. Insects have a low feed conversion ratio (kg feed/kg product) and can be consumed completely, without residual materials, such as bones or feathers. The nutritional value of insects is high, especially as a protein source for livestock. Insect-based feed products, therefore, can replace conventional feed ingredients, such as SBM. Altogether this strategy suggests that waste-fed larvae meal might become an important alternative feed source in the future. To gain insight into the status quo of the environmental impact of both mitigation strategies, replacing SBM with RSM or with waste-fed insects, we first used the attributional life cycle assessment (ALCA) method. Based on the ALCA method, results showed that each mitigation strategy was promising. Replacing SBM with RSM in growing pig diets hardly changed either global warming potential (GWP) or energy use (EU), but decreased land use (LU) up to 16% per kg body weight gain. As expected, feed production had the largest environmental impact, responsible for about 50% of the GWP, 60% of the EU, and 77% of the total LU. Feed production in combination with feed intake, were the most sensitive parameters; a small change in both these two parameters changed the results. Replacing SBM with waste-fed larvae meal in growing-pig diets showed that EU hardly changed, but GWP (29%) and LU (54%) decreased per kg body weight gain. Based on ALCA results, each mitigation strategy, therefore, seems to offer potential to reduce the environmental impact of pork production. An ALCA, however, has two disadvantages: it does not account for product-packages and it does not consider feed-food competition. The first disadvantage of ALCA was that the complexity of dealing with product-packages is not fully considered. ‘Product-package’ refers to a multiple-output situation. During the processing of sugar beet, for example, beet-pulp and molasses are produced in addition to sugar. Sugar, beet-pulp, and molasses together form a ‘package of products’ because they cannot be produced independently from each other. An ALCA does not account for the fact that the production volume of the co-product(s) depends on the demand for the determining product (e.g. sugar), which results in the limited availability of co-products. Increasing the use of co-products in animal feed, consequently, results in reducing use of a co-product in another sector, requiring them to be replaced with a different product. The environmental impact of increasing the use of a co-product or food-waste, therefore, depends on the net environmental impact. The net environmental impact refers to the environmental benefits of using the product in its new application minus the environmental cost of replacing the product in its old application. A consequential theoretical framework was developed to account for product-packages. The results, based on the consequential framework, contradicted standard ALCA results. The consequential LCA (CLCA) method we used for replacing SBM with RSM showed an increased GWP (up to 15%), EU (up to 12%), and LU (up to 10%) per kg body weight gain. Moreover, this CLCA method showed that replacing SBM with waste-fed larvae meal increased GWP (60%) and EU (90%), but decreased LU (73%) per kg body weight gain. Accounting for product-packages increased the net environmental impact of each strategy, replacing SBM with RSM or with waste-fed larvae meal. The difference in results between ALCA and CLCA was especially large in the strategy with waste-fed larvae meal. The difference was caused mainly by the use of food-waste. Food-waste fed to larvae was used initially to produce bio-energy via anaerobic digestion. In CLCA, the environmental impact related to replacing the bio-energy function of food-waste with fossil-energy was included. The net environmental impact became negative, because environmental benefits of replacing SBM with waste-fed larvae meal were less than environmental costs related to the marginal energy source, i.e. fossil energy, replacing the bio-energy. Results of the indirect environmental impact, however, are situation specific: if the marginal energy source were wind or solar energy, the net environmental impact of using waste-fed larvae meal might be positive. Waste-fed larvae meal, therefore, appears to be an interesting mitigation strategy only when energy from wind and solar energy are used more dominantly than energy from fossil sources. If results were based solely on ALCA, then these potentially negative impacts would have been overlooked. Consideration of the environmental consequences of product-packaging, therefore, is essential to determine total environmental costs. If policy makers or the feed industry want to assess the net environmental impact of a potential mitigation strategy, then we recommend to perform a CLCA instead of an ALCA. The framework developed in this thesis can be used to perform such an assessment. The second disadvantage of an LCA was that it does not take into account feed-food competition, e.g. competition for land between humans and animals. Most LCA studies focus on the total amount of land required to produce one kg ASF. LCA studies do not account for competition for land between humans and animals, or so-called feed-food competition. In other words, they do not include, differences in the consumption of human-edible products by various livestock species or differences in the suitability of land used for feed production as land to cultivate food-crops directly. Given the global constraints on land, it is more efficient to grow food directly for human consumption rather than for livestock. To address the contribution of livestock to a future sustainable food supply, a measure for land use efficiency was developed, called the land use ratio (LUR). The LUR accounts for plant productivity, efficiency of converting human-inedible feed into ASF, and suitability of land for crop cultivation. The LUR also has a life-cycle perspective. Results of the LUR illustrated that dairy cows on sandy soil, laying hens, and pig production systems in the Netherlands have a LUR >1.0. In terms of protein produced per m2, therefore, it is more efficient to use these soils for livestock production to produce crops for direct human consumption than to produce feed for livestock. Only dairy cows on peat soil produce human digestible protein (HDP) more efficiently than crops do, because peat is not suitable for crop production. The LUR allows identification of livestock production systems that are able to produce HDP more efficiently than crops do. Livestock systems with a LUR

The increasing world population and per capita income imposes a risk for protein scarcity. It is, therefore, necessary to use current ingredients more efficiently which includes the accurate assessment of protein quality before inclusion in animal diets. Protein quality is defined in this thesis as the capacity of a dietary protein to meet a pig’s requirement for nitrogen (N) and amino acids (AA) to meet a particular production target. Protein quality is influenced by processing applied to feed ingredients which may lead to the formation of Maillard reaction products (MRP) or cross-link products. The Maillard and cross-link reactions mainly involve lysine (Ly)s and their products may decrease ileal crude protein (CP) digestibility. During the acid hydrolysis step used to analyze AA, part of the early MRP revert back to Lys. This reverted Lys is not bioavailable for animals. Therefore, methods that specifically analyze Lys with a free ε-amino group (that is, not bound to other nutrients) have been developed. The guanidination reaction with O-methylisourea (OMIU) is one such method. The initial aim of this thesis was to evaluate the ileal digestible reactive Lys assay as a more accurate measure for protein quality of processed protein sources than the ileal digestible total Lys assay. Soybean meal (SBM) and rapeseed meal (RSM) were used as sole protein sources throughout this thesis. Processing of SBM and RSM by toasting at 95°C for 30 min in the presence of a sugar-rich lignosulfonate was used as model for over-processed protein sources. Digestibility, post-absorptive utilization, and pig growth performance In Chapter 2, protein quality in processed protein sources was determined using the content of AA, OMIU-reactive Lys, MRP, and lysinoalanine (LAL; as cross-link product), the standardized ileal digestibility (SID) of AA and OMIU-reactive Lys and pig growth performance. The SBM and RSM diets contained furosine and carboxymethyllysine (CML) as MRP, and LAL indicating that the Maillard and cross-link reactions had taken place in SBM and RSM, presumably during the oil extraction/desolventizing process. The amounts of furosine, CML, and LAL were elevated in the pSBM and pRSM diets due to further processing. Processing resulted in a reduction in total and OMIU-reactive Lys contents, a decreased pig growth performance as determined by the gain to feed ratio (G:F), and the SID of CP, AA, and OMIU-reactive Lys. The SID AA contents of the protein sources from Chapter 2 were used to formulate the diets of the main in vivo experiment (Chapters 3 and 4). In this experiment, six experimental diets were used of which four contained either SBM, pSBM, RSM, or pRSM as sole protein source. The remaining two experimental diets contained pSBM or pRSM and were supplemented with crystalline AA to the same SID AA levels as the SBM or RSM diet. These supplemented diets were used to verify that processing affected AA digestibility rather than post-absorptive AA utilization. The effects of processing on CP digestibility and N solubilization along the small intestine, metabolic load as assessed by organ weight, and nutrient composition of the empty body of growing pigs are described in Chapter 3. The small intestine was divided in three segments of similar length and digesta was collected from the last 100 cm of each segment. The amount of insoluble N as a fraction of N in digesta at each small intestinal segment was not affected by processing. Thus, the reduced SID of CP and AA reported in Chapter 2 was not caused by a reduced N solubility but by a general increase of N in digesta. Processing reduced the SID of CP, CP content in the empty body, and G:F. Supplementing crystalline AA to diets containing pSBM or pRSM increased the CP content and G:F to the level of the SBM and RSM diets. Processing also reduced the weight of several organs and supplementing crystalline AA restored organ weight. The effects of processing on whole body AA composition, nutrient retention, and post-absorptive utilization of AA in growing pigs are described in Chapter 4. Post-absorptive AA utilization was calculated as percentage of SID AA intake used for AA retention. Processing affected the AA composition of protein in the organ fraction (that is, empty organs and blood), carcass, and empty body. The Lys concentration in body protein was mainly reduced by processing. Supplementing crystalline AA restored the AA composition of body protein for SBM and RSM. Processing reduced AA retention and again supplementing crystalline AA restored AA retention for both SBM and RSM. Since crystalline AA were supplemented on an SID AA basis, the results indicated that processing affected AA digestibility but not post-absorptive AA utilization. Thus, correcting AA retention for SID AA intake would result in a similar post-absorptive AA utilization which was found for most AA for the RSM diets. However, the post-absorptive AA utilization was lower for the pSBM diet than for the SBM diet which might be related to an imbalanced AA supply after absorption in the first diet. The assessment of ileal digestibility and utilization is expensive and laborious. Therefore, two alternative in vitro methods for determining protein digestibility for processed protein sources were evaluated (Chapter 5). The protein digestibility determined using the pH-STAT method and a 2-step enzymatic method was compared with the in vivo SID of CP reported in Chapter 2. Initial pH and the degree of hydrolysis assessed in the pH-STAT method were positively correlated to SID of CP. Protein digestibility determined with the 2-step enzymatic method, simulating digestion in the stomach and small intestine, tended to correlate to SID of CP. Both the 2-step enzymatic method and pH-STAT method were suitable alternatives for the assessment of SID of CP. However, only four ingredients were tested. The suitability of the methods should be further studied using multiple (processed) feed ingredients before they can be used as alternatives for in vivo assays. Reactive Lys analysis O-methylisourea was reported to bind specifically to the ε-amino group of Lys. The results of Chapter 2, however, cast doubt on the specificity of OMIU to react only with the ε-amino group of Lys. A series of experiments was conducted to study this specificity (Chapter 6). Incubating crystalline L-Lys with OMIU under standard conditions (OMIU pH of 10.6, OMIU to AA ratio of 1000:1, and reaction time of 7 d) resulted in a low homoarginine (that is, Lys with OMIU bound to its ε-amino group) recovery. The reaction of OMIU with the α-amino group of Lys was confirmed by mass spectrometry analysis with double derivatized Lys being identified. Several reaction conditions (OMIU pH, OMIU to Lys ratio, and reaction time) were studied but none of these resulted in 100% recovery of homoarginine. Binding of OMIU to the α-amino group of Lys could result in an underestimation of the reactive Lys content when significant levels of Lys with a free α-amino group (that is, crystalline L-Lys (HCl), free and N-terminal Lys) are present in food/feed ingredients, diets, and ileal digesta. The free Lys content in food/feed ingredients was on average 1.3% of total Lys. The free Lys content can be substantial in certain diets and was reported to be 13% of total Lys in ileal digesta. The latter might result in an overestimation of the OMIU-reactive Lys digestibility. The reaction of OMIU with α-amino groups may necessitate analysis of free Lys to accurately quantify reactive lysine in samples containing a large proportion of Lys with a free α-amino group. The results presented in this thesis indicate that the effects of processing on SID of CP and AA, body composition, nutrient retention, post-absorptive AA utilization, and growth performance could be substantial. These effects should, therefore, be taken into account when using processed feed ingredients in diets for growing pigs. The extent of protein damage in feed ingredients can be assessed by the analysis of OMIU-reactive and total Lys, MRP, and cross-link products. However, OMIU-reactive Lys only provides accurate results when samples contain small levels of Lys with a free α-amino group (that is, crystalline L-Lys (HCl), free and N-terminal Lys). When samples contain significant levels of Lys with a free α-amino group, it is recommended to use standard guanidination conditions (OMIU pH of 10.6, OMIU to AA ratio of 1000:1, and reaction time of 7 d) to convert protein-bound Lys to homoarginine and to separately analyze such samples for free Lys.