xx, 65 leaves : graphs ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Agricultural Biochemistry, 1979

Pharmacological levels of inorganic forms of copper (Cu) and zinc (Zn) are frequently used in diets for pigs to improve performance and control post-weaning colibacillosis. However, the use of such forms and levels causes mineral accumulation in the soil, and is a non-sustainable practice from an environmental perspective. Alternatively, organic complexes of copper (Cu) and zinc (Zn) have been proposed to be more available to pigs, and when included at lower levels than inorganic sources of these minerals have significantly reduced mineral excretion in faeces without compromising performance. However the effect of these organic minerals fed simultaneously at low levels of inclusion has not been well studied. The general hypothesis tested in this thesis was that concentrations of Cu and Zn in faecal material would be reduced when fed in an organic (Bioplex®) form without compromising performance or mineral homeostasis in growing/finishing pigs, in comparison to Cu and Zn fed in an inorganic (sulphate) form. Two experiments were performed to test this hypothesis: Experiment 1 was designed as a 2x2 factorial arrangement of treatments, with two mineral forms (Bioplex® and Sulphate) and two inclusion levels (High and Low). The “low” levels aimed at providing 80 mg/kg of dietary Cu and Zn, and the “high” levels aimed at providing 160 mg/kg of dietary Cu and Zn. Experiment 2 was designed as a 3x2 factorial arrangement of treatments, with two mineral forms (Bioplex® and Sulphate) and three inclusion levels (Low, Medium and High). The “low” levels aimed at providing 25 mg/kg of Cu and 40 mg/kg of Zn, the “medium” levels aimed at providing 80 mg/kg of both Cu and Zn, and the “high” levels aimed at providing 160 mg/kg of both Cu and Zn in the diet. Unfortunately the Medium sulphate grower diet was contaminated with excess Zn while manufacturing, which led to the exclusion of this treatment from the study. In Experiment 1, pigs fed LB (Low Bioplex®) or HS (High Sulphate) diets grew faster (P=0.014) and their carcasses were 3.5 kg heavier (P=0.020) than LS (Low Sulphate)- or HB (High Bioplex®)-fed pigs. Pigs fed LB or HS diets had lower (P=0.001) levels of Zn in plasma, a higher (P=0.029) concentration of Zn in the pancreas and a lower (P=0.020) concentration of Zn in bone than pigs fed LS or HB diets. The concentration of Cu in liver increased (P=0.017) with the concentration in the diet as did Cu and Zn levels in faeces (P<0.001) without any difference between mineral forms. Feed conversion ratio (FCR) tended to be improved (P=0.062) by the inclusion of Bioplex® in the diet. The inclusion of Bioplex® reduced (P=0.003) subcutaneous fat depth at the P2 site by 2.2 mm compared to the sulphate. In Experiment 2, there was no difference (P>0.05) in growth rate between experimental diets, but again there was an overall improvement (P=0.012) in FCR when Bioplex® were included. Blood and tissue Cu and Zn concentrations were within normal physiological ranges in all treatments, supporting a reduction of Cu and Zn levels in the diet. Only Zn level in plasma during the growing phase and Cu and Zn concentration in tissues increased (P<0.001) with the addition in the diet. None of the biomarkers of Cu or Zn status analysed in the pigs showed any difference between the inorganic and the Bioplex® forms. Copper and Zn concentrations in faecal material decreased (P<0.001) with their inclusion in the diet, and only in the finishing collection there was a further decrease of 10% in Zn faecal concentration when Bioplex® was included instead of the sulphate at similar low levels. Carcass and meat quality measures were independent of the Cu and Zn form or level, however a higher proportion of carcasses from LB-fed pigs had <14 mm subcutaneous fat depth at the P2 site. The inclusion of Bioplex® failed to have a significant effect on Cu excretion and its inclusion had an inconsistent effect on Zn excretion. The overall findings from this thesis partially supported the hypothesis that the inclusion of Bioplex® would reduce the concentration of Cu and Zn in faeces compared to the inclusion of inorganic forms at similar inclusion levels. Nevertheless, total Cu and Zn levels in growing/finishing pig diets could be reduced from 160 mg/kg of both Cu and Zn to 30 mg/kg Cu and 60 mg/kg Zn, in either the sulphate or the Bioplex® form, without negatively affecting performance or mineral homeostasis in the pigs and significantly reducing Cu and Zn excretion (between 50 and 80%). The advantage of including Bioplex® instead of sulphates was in the improvement in FCR. Carcass and meat quality were independent of the form and level of dietary Cu and Zn.

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Dans un contexte de développement durable, trouver des moyens de réduire l’impact environnemental de la production avicole est d’une grande importance. L’estimation des paramètres génétiques de nombreux caractères de rejets chez les lignées divergentes D+ et D- nous a permis de définir plusieurs critères pertinents à prendre en compte dans les schémas de sélection pour maîtriser l’excrétion des poulets. Nous avons également montré que l’utilisation de ces critères n’aurait pas de conséquences négatives sur les caractéristiques d’importance économique pour l’industrie avicole. Enfin, nous avons pu montrer que la morphologie du tractus gastro-intestinal était très fortement impliquée dans les variations d’excrétion des poulets. En effet, un important développement du proventricule et du gésier et une réduction du développement de l’intestin grêle sont associés aux animaux produisant moins de rejets et présentant une meilleure efficacité digestive. / The reduction of the environmental impact of poultry production is crucial to ensure its sustainability. The estimation of genetic parameters of several excretion characteristics in D+ and D- divergent lines allowed us to define pertinent criteria that could be included in selection schemes in view to reduce chicken excretion. We also showed that using these criteria would not have negative consequences on the traits of economic importance for poultry industry. Finally, we showed that the gastro-intestinal tract morphology is highly implied in the variations of excretion between chickens. Indeed, an important development of the proventriculus and gizzard and a reduction of the small intestine development were associated with birds producing less excreta and showing a higher digestive efficiency.

[ES] Los estudios sobre nutrición proteica en conejos son escasos en comparación con otras especies, el cálculo de las necesidades es relativamente antiguo, no están ofrecidos a nivel ileal y sus peculiares hábitos alimenticios complican su conocimiento. Además, en los últimos años, se ha establecido relación directa entre este nutriente con una de las principales enfermedades que afectan esta especie (la Enteropatía Epizoótica del Conejo), y como consecuencia, el contenido proteico de las dietas se ha reducido. Por otro lado, se ha comprobado una pérdida en la efectividad de la selección por velocidad de crecimiento en las líneas paternales la cual podría estar relacionada con una mala estimación de los requerimientos proteicos (especialmente en los animales con elevadas velocidades de crecimiento). Finalmente, el nitrógeno ureico plasmático (PUN) podría ser una buena herramienta para estimar el grado de aprovechamiento proteico del pienso, pero su uso no ha sido comprobado aún en esta especie. Con todo ello, la principal hipótesis de esta tesis es que existe una mala estimación de las necesidades de aminoácidos en conejos con elevadas velocidades de crecimiento y que otra combinación de aminoácidos podría maximizar su rendimiento. Para evaluar esta hipótesis se utilizaron un total de 1464 conejos de engorde (provenientes principalmente de la línea R, línea paternal seleccionada por velocidad de crecimiento, y, en mucha menor medida, de líneas maternales con menores velocidades de crecimiento), se formularon un total de 32 dietas (donde principalmente diferían el contenido de los tres aminoácidos típicamente limitantes; lisina, aminoácidos azufrados y treonina), se realizaron 69 y 71 ensayos de digestibilidad fecal e ileal, respectivamente, se valoró la retención de nutrientes de 126 animales durante su crecimiento y por último se analizaron los niveles de PUN de más de 2700 muestras. Con el primero de los trabajos obtuvimos la confirmación del primer indicio: las dietas actuales parecían presentar deficiencias en aminoácidos para los conejos con altas velocidades de crecimiento, comprobando que aquellos animales que crecían más depositaban proporcionalmente más energía y menos proteína (esto sugería una desviación de cierta proporción de proteína como fuente de energía, siendo poco eficiente). Antes de plantearnos qué combinación de aminoácidos mejoraría este problema, tendríamos que poner a punto la técnica para detectar dichas deficiencias. En el segundo trabajo no solo comprobamos que el PUN podría ser utilizado, en esta especie, sino que se estableció la mejor metodología: A las 08:00h (bajo una alimentación ad libitum) o a las 21:00h (bajo un ayuno de 10h y un restablecimiento del alimento a las 18:00h). Ahora ya, sabiendo que podría existir dicha deficiencia y que además podemos utilizar el PUN, seleccionamos de una matriz de 27 piensos (donde habían tres niveles de lisina, azufrados y treonina), aquel que optimizaba los valores de PUN y que previsiblemente mejoraría la asimilación proteica. De este experimento surge la Dieta MAB (8.1, 6.6 y 5.7 g/kg materia seca de lisina, aminoácidos azufrados y treonina, respectivamente) como candidato a sustituir a las actuales recomendaciones, Dieta MMM (8.1, 5.8 y 6.9 g/kg materia seca de lisina, aminoácidos azufrados y treonina, respectivamente). Pero hasta ahora solo podíamos decir que el MAB reduce los nivele de urea: ¿Está esto relacionado con unos mejores parámetros productivos? Finalmente en el último trabajo se comprobó la superioridad productiva de esta combinación, ya que mejoró significativamente tanto la velocidad de crecimiento como el índice de conversión. Tras la valoración digestiva, podemos aconsejar unos niveles de 5.3, 4.3 y 2.9 g/kg de materia seca para la lisina, aminoácidos azufrados y treonina a nivel ileal aparente, arrojando más información sobre la nutrición proteica de precisi� / [CAT] Els estudis sobre nutrició proteica en conills són escassos en comparació amb altres espècies, el càlcul de les necessitats és relativament antic, no estan oferts a nivell ileal i els seus peculiars hàbits alimentaris compliquen el seu coneixement. A més, en els últims anys, s'ha establert relació directa entre aquest nutrient amb una de les principals malalties que afecten aquesta espècie (la enteropatia epizoótica del Conill), i com a conseqüència, el contingut proteic de les dietes s'ha reduït. D'altra banda, s'ha comprovat una pèrdua en l'efectivitat de la selecció per velocitat de creixement en les línies paternals la qual podria estar relacionada amb una mala estimació dels requeriments proteics (especialment en els animals amb elevades velocitats de creixement). Finalment, el nitrogen ureic plasmàtic (PUN) podria ser una bona eina per estimar el grau d'aprofitament proteic del pinso, però el seu ús no ha estat comprovat encara en aquesta espècie. Amb tot això, la principal hipòtesi d'aquesta tesi és que hi ha una mala estimació de les necessitats d'aminoàcids en conills amb elevades velocitats de creixement i que una altra combinació d'aminoàcids podria maximitzar el seu rendiment. Per avaluar aquesta hipòtesi es van utilitzar un total de 1464 conills d'engreix (provinents principalment de la línia R, línia paternal seleccionada per velocitat de creixement, i, en molta menor mesura, de línies maternals amb menors velocitats de creixement), es van formular un total de 32 dietes (on principalment diferien el contingut dels tres aminoàcids típicament limitants; lisina, aminoàcids ensofrats i treonina), es van realitzar 69 i 71 assajos de digestibilitat fecal i ileal, respectivament, es va valorar la retenció denutrients de 126 animals durant el seu creixement i finalment es van analitzar els nivells de PUN de més de 2700 mostres. Amb el primer dels treballs vam obtenir la confirmació del primer indici: les dietes actuals semblaven presentar deficiències en aminoàcids per als conills amb altes velocitats de creixement, comprovant que aquells animals que creixien més dipositaven proporcionalment més energia i menys proteïna (això suggeria una desviació de certa proporció de proteïna com a font d'energia, sent poc eficient). Abans de plantejar- nos què combinació d'aminoàcids milloraria aquest problema, hauríem de posar a punt la tècnica per detectar aquestes deficiències. En el segon treball no només vam comprovar que el PUN podria ser utilitzat, en aquesta espècie, sinó que es va establir la millor metodologia: A les 08: 00h (sota una alimentació ad libitum) oa les 21: 00h (sota un dejuni de 10h i a un restabliment de l'aliment a les 18: 00h). Ara ja, sabent que podria existir aquesta deficiència i que a més podem utilitzar el PUN, seleccionem d'una matriu de 27 pinsos (on havien tres nivells de lisina, ensofrats i treonina), aquell que optimitzava els valors de PUN i que previsiblement milloraria l'assimilació proteica. D'aquest experiment sorgeix la Dieta MAB (8.1, 6.6 i 5.7 g / kg matèria seca de lisina, aminoàcids ensofrats i treonina, respectivament) com a candidat a substituir les actuals recomanacions, Dieta MMM (8.1, 5.8 i 6.9 g / kg matèria seca de lisina, aminoàcids ensofrats i treonina, respectivament). Però fins ara només podíem dir que el MAB redueix els anivelli d'urea: Està això relacionat amb uns millors paràmetres productius? Finalment en l'últim treball es va comprovar la superioritat productiva d'aquesta combinació, ja que va millorar significativament tant la velocitat de creixement com l'índex de conversió. Després de la valoració digestiva, podem aconsellar uns nivells de 5.3, 4.3 i 2.9 g / kg de matèria seca per la lisina, aminoàcids ensofrats i treonina a nivell ileal aparent, llançant més informació sobre la nutrició proteica de precisió i pretenent tallar la problemàtica que ens plantejàvem a l'inici de la tesi. / [EN] Studies on protein nutrition in rabbits are scarce compared with other species, the estimation of their requirements is relatively old and no data are reported at ileal level. Moreover, their peculiar feeding behaviour, including caecotrophy, complicates more this study. In addition, in recent years, a relationship has been established between dietary crude protein and Epizootic Rabbit Enteropathy (one of the most important disease that affects this species). As a consequence, the protein content of diets has been reduced. On the other hand, there has been a loss of effectiveness of the selection for growth rate in the paternal lines, which could be related with a possible nutritional deficit in high growth rate rabbits when using the current commercial diets. Plasma urea nitrogen (PUN) could be a good indicator of an amino acid imbalance, but its use has not yet been tested in rabbits. Thus, the main hypothesis of this thesis concerns that growing rabbits selected by growth rate could have amino acid requirements different from current recommendations, and that another amino acid combination would maximize productive traits. To evaluate this hypothesis, a total of 1464 growing rabbits were used (mainly from the R line, a paternal line selected by growth rate, and, to a much lesser extent, from some other maternal lines with lower growth rates), 32 diets were formulated differing in the content of the three typically limiting amino acids (lysine, sulfur amino acids and threonine). Faecal and ileal digestibility trials were performed with 69 and 71 animals, respectively. Nutrient retention during their growing period was evaluated in 126 animals. Finally, PUN was analyzed in more than 2700 samples. From the first trial,we obtained indication about the possible existence of some limiting amino acid when current moderate protein diets are used in growing rabbits with high growth rates, as retained protein to retained energy ratio was clearly lower for animals of the paternal lines (this suggested a deviation of a certain proportion of protein as a source of energy, which is inefficient). Before considering what combination of amino acids would improve their utilization, it was necessary to evaluate the possible interest of PUN to detect deficiencies in amino acids and establish the appropriate methodology to optimize its use for this purpose in growing rabbits. In the second study we verified the applicability of PUN in rabbits and checked the highest differences between diets: from 04:00h to12:00h in animals fed ad libitum and at three hours after refeeding (21:00h) when a fasting period of 10 h was applied. At this point, knowing that amino acid deficiency could exist and that PUN could be an indicator of it, we formulated a matrix of 27 diets (in a 3x3x3 factorial design, with three levels of the three amino acids mentioned above, using the current recommendations and increasing or decreasing them by 15%), searching for the combination minimizing PUN values and, hypothetically, improving the protein assimilation. From this trial, the diet MHL (8.1, 6.6 and 5.7 g/kg dry matter of lysine, sulphur amino acids and threonine, respectively) emerged as a candidate to replace the current recommendations (diet MMM; 8.1, 5.8 and 6.9 g/kg dry mater of lysine, sulphur amino acids and threonine, respectively). However a question arises: could this new combination improve productive traits? Finally, in the last study we verified the usefulness of this combination since it significantly improved both the growth rate and the feed conversion ratio. After digestibility assessment, we propose levels of 5.3, 4.3 and 2.9 g/kg dry matter of apparent ileal digestible lysine, sulphur amino acids and threonine, respectively. These data are intended to improve current knowledge on protein nutrition and move towards precision protein nutrition. / Marín García, PJ. (2019). Lysine, sulfur amino acids and threonine requirements of growing rabbits from a line selected by growth rate [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/132186 / TESIS

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京都大学 / 新制・課程博士 / 博士(人間健康科学) / 甲第23824号 / 人健博第95号 / 新制||人健||7(附属図書館) / 京都大学大学院医学研究科人間健康科学系専攻 / (主査)教授 十一 元三, 教授 林 悠, 教授 小林 恭 / 学位規則第4条第1項該当 / Doctor of Human Health Sciences / Kyoto University / DFAM

Protein has long been recognized as an essential dietary substance, necessary for the building and repair of body tissue. However, in recent years we have learned even more of the versatility of protein in body processes. In addition to the need of protein for growth and repair of tissue breakdown, protein compounds are now known to serve also as parts of hormones to regulate body processes, enzymes to digest foods, plasma albumin to maintain water balance and blood volume, globulins to resist infection, and hemoglobin to transport oxygen to the tissues. In times of food shortages, protein deficiency is more likely to occur than that of any other of the important dietary essentials. This is due to the fact that there is a shortage of the foods that furnish protein of high biological value, i.e., the proteins of meat, milk, eggs and cheese. At the present time, where food shortages exist, these foods are scarce and when available expensive. Because of this situation, research concerned with protein metabolism is especially significant today. Proteins differ from one another on the basis of the number and kind of amino acids of which they are made. Since there are nine (possibly 10) essential amino acids, that cannot be synthesized in the body in adequate amounts from other amino acids, they must be present in the food. These essential amino acids are found in protein of animal origin. Plant protein is good for supplementing other protein, but alone, it is inadequate for maintenance and growth in the body. With complete protein foods both scarce and expensive, a knowledge of the protein requirement, the factors that affect its utilization, and how the body needs are met are of prime importance. The protein requirement has been determined from the data obtained from protein (or nitrogen) balance experiments in man. However, due to the nature of the data and the many variable factors involved in the amino acid content of various dietaries, there have been disagreements as to the real requirement, some advocating a high intake, others suggesting a lower intake. Today a compromise of the two extremes is generally advocated. The National Research Council has suggested an adult protein allowance of 60 to 70 gm. per day, from a diet of mixed protein (both animal and plant foods). In view of the fact that the number of studies to determine the protein requirement of human beings has of necessity been small, this problem was originally undertaken to observe the effect of the variation of caloric intake on protein use by means of the nitrogen balance experiment. In developing a low protein diet for the above study, coffee was suggested by one of the subjects to make the diet more palatable. This raised the question of whether or not coffee might in some way affect protein utilization or nitrogen excretion. A search was made for data on this point. In reviewing the available literature it was found that workers between 1853 and 1912 had done some work on the effect of caffeine on the nitrogen in the urine of men and dogs, but their results were contradictory, and in most cases they had used varying and unspecified amounts of caffeine. Since that time this problem has apparently been neglected. In the hope of obtaining further information concerning the effect of coffee on nitrogen metabolism, this study was planned with the following purposes: To 1. Develop dietary and laboratory technique for the nitrogen balance experiment. 2. Attempt to reach the point of nitrogen equilibrium in subjects on a known protein and caloric intake. 3. Determine the effect, if any, of the ingestion of a known amount of coffee on the total nitrogen, urea and ammonia excretions. / Master of Science