Lo scopo del presente lavoro è stato quello di esaminare il potenziale di idrolisi e degradabilità dell’amido dei cereali e co-prodotti e di valutare gli aspetti collegati. L’approccio utilizzato comprende sia test in vitro che in vivo. In particolare: - Una serie d’indagini in vitro ha fornito dati sul potenziale di degradabilità enzimatica dell’amido dei cereali e co-prodotti considerando sia l’influenza della struttura dell’amido, del tipo di cereale, del rapporto amilosio:amilopectina, del processo termico, dell’insilamento, dello stadio di maturazione e delle pratiche agronomiche. - Una valutazione funzionale basata sul calcolo del potenziale di degradabilità enzimatica dell’amido tramite la stima dell’indice glicemico in vitro, combinata con un approccio matematico di modellizzazione dei dati ottenuti, è stata proposta per fornire un metodo efficace di screening dei materiali amidacei utilizzati comunamente nella formulazione per diete destinate ai suini. - Un esperimento in vitro è stato condotto per investigare la risposta glicemica post-prandiale in suini alimentati con diete che differivano nel potenziale di idrolisi enzimatica. Infine, considerando la crescente attenzione dei nutrizionisti sulla matrice proteica dei cereali (con particolare importanza data alle prolammine), i metodi comunemente utilizzati per le tecniche di estrazione delle prolammine sono stati confrontati e ri-investigati. / In the present work, it has been attempted to examine the variability in starch hydrolysis and digestion from cereal grains and co-products and to study aspects related, in order to give a better understand of the science behind the differences in grains/feeds starch digestion potential. The general approach was to use a combination of in vitro and in vivo techniques. In particular: - A series of in vitro investigations has provided data on the degradation potential of starch from cereal grains and co-products considering the influence of starch structure, cereal type, amylose-amylopectin ratio, heat processing, ensiling, stage of maturity and of agricultural practices. - A laboratory-based functional evaluation of starch value of cereal grains based on a predicted glycemic index approach, combined with a mathematical first-order exponential model, has been proposed in order to allow an efficient screening of starchy-materials entering in the pig diet formulation. - An in vivo experiment has been undertaken to investigate the postprandial plasma glucose response in pigs fed diets containing starch with a wide range in in vitro digestion patterns. Lastly, taking into account the increasing attention of swine nutritionists on prolamin proteins, the most commonly used prolamin extraction procedures were compared and reinvestigated, in order to get useful information for routine lab analysis.

La present Tesi Doctoral s’emmarca dins d’una línia d’investigació dedicada a l’estudi de les bases genètiques del metabolisme dels greixos en relació a la producció de carn de porcí d’alta qualitat i saludable. L’objectiu final és la identificació de polimorfismes i mecanismes de regulació responsables de la variabilitat genètica d’aquests caràcters complexes en l’espècie porcina. En aquest context, s’han utilitzat mètodes de genòmica estructural (mapes de QTL d’expressió (eQTL); estudis de gens candidats) i funcional (estudis d’expressió gènica) a més a més d’anàlisis proteics i cel·lulars en mostres de múscul i greix de porcs seleccionats per les seves característiques de qualitat de carn. Així, mitjançant la tècnica de microarrays s’ha analitzat el patró d’expressió d’ARNm en mostres de múscul gluteus medius obtingudes a partir de porcs d’una població comercial Duroc amb fenotips divergents per diversos paràmetres relacionats amb la deposició dels lípids. Com a resultat, s’han observat nombrosos gens diferencialment expressats entre els animals amb perfils divergents d’engreixament. Un estudi ontològic/funcional va revelar que aquests gens estaven particularment relacionats amb el metabolisme lipídic, el creixement i la diferenciació muscular, la immunitat i la captació de glucosa en la ruta de la insulina. D’altra banda, les anàlisis d’eQTL han revelat l’existència de regions genòmiques responsables de la variació de l’expressió gènica en el múscul gluteus medius porcí; algunes de les quals mostren una concordança posicional amb varis QTL per caràcters de qualitat de cran i engreixament detectats prèviament a la mateixa població Duroc. Complementàriament, s’ha realitzat un estudi més exhaustiu de quatre gens candidats (ACACA, HMGCR, SCD i 6D), directament implicats en caràcters relacionats amb la qualitat de la carn al ser els principals responsables de la síntesis i dessaturació d’àcids grassos i colesterol. Combinant els resultats de l’anàlisi d’expressió gènica, mapes d’eQTL i els gens candidats estudiats s’ha elaborat una llista de gens candidats funcionals i posicionals que serà la base de futures investigacions cap a l’establiment de les xarxes gèniques i els mecanismes moleculars implicats en el metabolisme dels lípids musculars i els caràcters relacionats amb la qualitat de la carn en porcí. / La presente Tesis Doctoral se enmarca en una línea de investigación dedicada al estudio de las bases genéticas del metabolismo de las grasas en relación a la producción de carne de porcino de alta calidad y saludable. El objetivo final es la identificación de polimorfismos y mecanismos de regulación responsables de la variabilidad genética de estos caracteres complejos en porcino. Para ello se han utilizado métodos de genómica estructural (mapas de QTL de expresión (eQTL); estudio de genes candidatos) y funcional (estudios de expresión génica) y también de análisis proteico y celular en muestras de músculo y grasa de cerdos seleccionados por sus características de calidad de carne. Así, mediante la técnica de microarrays se ha analizado el patrón de expresión de ARNm en muestras de músculo gluteus medius obtenidas a partir de cerdos de una población comercial Duroc con fenotipos divergentes para varios parámetros relacionados con la deposición de los lípidos. Como resultado, se han observado numerosos genes diferencialmente expresados entre los animales con perfiles divergentes de engorde. Un estudio ontológico/funcional mostró que estos genes estaban particularmente relacionados con el metabolismo lipídico, el crecimiento y la diferenciación muscular, la inmunidad, y la captación de glucosa en la ruta de la insulina. Por otra parte, un análisis de eQTL ha revelado la existencia de regiones genómicas responsables de la variación de la expresión génica en el músculo gluteus medius porcino, algunas de las cuales muestran una concordancia posicional con varios QTL para caracteres de calidad de carne y engorde detectados previamente en la misma población Duroc. Complementariamente, se ha realizado un estudio más exhaustivo de cuatro genes candidatos (ACACA, HMGCR, SCD y 6D) directamente implicados en caracteres relacionados con la calidad de la carne al ser los principales responsables de la síntesis y desaturación de ácidos grasos y colesterol. Combinando los resultados del análisis de expresión génica, mapas de eQTL y los genes candidatos estudiados se ha elaborado una lista de genes candidatos funcionales y posicionales que será la base de futuras investigaciones hacia el establecimiento de las redes génicas y los mecanismos moleculares implicados en el metabolismo de los lípidos musculares y los caracteres relacionados con la calidad de la carne en porcino. / This PhD is part of a line of research devoted to studying the genetic basis of lipid metabolism and fat deposition in pigs with a view to producing healthy and high quality meat. The main objective is the identification of polymorphisms and regulatory mechanisms responsible for the genetic variability of these complex characters in pigs. In this sense, we have used several methods in the fields of structural (expression QTL (eQTL) maps; candidate genes studies) and functional (gene expression studies) genomics and also protein and cell studies in muscle and fat samples from pigs selected by meat quality parameters. In this context, using microarrays we analyzed the mRNA expression pattern in gluteus medius muscle samples obtained from a commercial Duroc pig population with divergent phenotypes for several parameters related to lipid deposition. As a result, we have obtained a list of genes differentially expressed between animals with divergent profiles related to lipid deposition. The ontological/functional study showed that these genes were particularly related to lipid metabolism, growth and muscle differentiation, immunity and glucose uptake in the insulin pathway. Moreover, analysis of eQTL has revealed the existence of genomic regions responsible for the variation of gene expression in porcine gluteus medius muscle. Some of these eQTL show positional concordance with several QTL related to meat quality and fat deposition previously identified in the same Duroc population. Additionally, we have performed a comprehensive study of four candidate genes (ACACA, HMGCR, SCD and 6D) directly involved in traits related to meat quality, playing an important role in fatty acids and cholesterol synthesis and desaturation. Combining the results of gene expression analysis, eQTL maps and candidate genes studied have resulted in a list of functional and positional candidate genes representing a valuable contribution to the understanding of the genetic regulation of skeletal muscle individual gene expression in swine species. This is a first step towards disentangling gene networks and molecular mechanisms involved in muscular lipid metabolism and meat quality traits in pigs.

El objetivo del presente trabajo consistió en valorar el efecto de las células epiteliales del oviducto porcino y del cumulus oophorus sobre distintos parámetros relacionados con la fecundación in vitro en la especie porcina. Las variables estudiadas fueron el porcentaje de penetración, el número medio de espermatozoides por ovocito, el porcentaje de formación de pronúcleo masculino, el porcentaje de monospermia, el porcentaje de cigotos divididos a las 48 horas postfecundación, el porcentaje de blastocistos, el número medio de núcleos por blastocisto, la resistencia de la zona pelúcida a la digestión con pronasa y la densidad y distribución de los gránulos corticales en los diferentes grupos experimentales. / The objective of this experimental work was to study the effect of porcine epithelial oviductal cells and cumulus cells on several parameters related to in vitro fertilization in pigs. Studied variables were rate of penetration, mean number of sperm cells per penetrated oocyte, rate of male pronuclear formation, cleavage rate at 48h postinsemination, blastocyst formation rate, mean number of cells per blastocyst, resistance of zona pellucida to pronase digestion and density and distribution of cortical granules in the different experimental groups.

Con el presente trabajo se ha pretendido investigar la influencia de diversos factores relacionados con el cocultivo de los gametos porcinos, sobre los resultados de la fecundación" in vitro" (AV), fundamentalmente con la intención de mejorar la eficacia actual del sistema de AV en cuanto a la consecución de embriones viables (fecundaciones monospérmicas). Para ello, se han utilizado 129 hembras porcinas prepúberes a las que se indujo la ovulación mediante un tratamiento con 1250 U.I. de PMSG y 750 U.I. de HCG. El tratameinto empleado resultó eficaz para los fines perseguidos en un 80'62% de las hembras y el número medio de ovocitos recogidos fue de 19'07 + 1'52 por animal, utilizándose un total de 1984 ovocitos.En relación a las condiciones del sistema de fecundación, el primer factor investigado ha sido el tiempo de cocultivo, entendido como tiempo de contacto entre los gametos. En las dos experiencias realizadas, utilizando tiempos de 4, 6 u 8 horas (experiencia la), o de 1, 2, 3 ó 4 horas (experiencia lb), los mejores resultados se obtuvieron tras 4 horas de cocultivo, ya que los porcentajes de penetración de mantuvieron altos con respecto al máximo alcanzado a las 8 horas (82,68 vs. 93,96%), mientras que los de monospermia no disminuyeron excesivamente con respecto a los obtenidos con tiempos de cocultivo menores, teniendo en cuenta que la concentración de espermatozoides empleada fue intencionadamente elevada (12 x 105 esp/ml). El segundo factor analizado fue la concentración espermática. Se utilizaron concentraciones de 3, 6 y 12 x 105 esp vivos/ml, deduciéndose de los resultados que la mayor efectividad en nuestro sistema correspondía a la concentración de 6 x lO 5 esp/ml, ya que los porcentajes de penetración fueron significativamente diferentes a los obtenidos con la concentración espermática más alta (71'62% vs. 76'83%), y los porcentajes de monospermia tampoco se diferenciaron de los obtenidos con la concentración espermática más baja (62'26 vs. 68'08%). El tercer factor estudiado ha sido la influencia de la presencia o ausencia en el medio de cocultivo del "cumulus" expandido que acompaña al ovocito en la ovulación. Por los resultados obtenidos, se puede pensar que la presencia estas células junto con la correspondiente matriz intrecelular de ácido hialúrico es altamente beneficiosa para la mejora del rendimiento de la FIV debido a que los porcentajes de penetración en los ovocitos denudados (53'69 fueron menores (p<0'0l) que los obtenidos en los ovocitos con "cumulus" (69'10%), mientras que los porcentajes de monospermia fueron superios (p < 0'01) en el segundo caso (39'45% vs. 60'97%). Por último, se ha investigado el efecto de la reducción del volumen medio de cocultivo más comúnmente utilizado (2 mI) a otro menor (0'4 n obteniéndose resultados equivalentes en ambos casos para los porcentajes penetración, pero mayores porcentajes de monospermia (p<0'05) con volumen de 0'4 mI (57'53% vs. 78'12%). Del conjunto de los resultados se deduce que los porcentajes de penetración y polispermia en la AV porcina son consecuencia de la influencia de diferentes factores, entre los que se encuentran el tiempo de cocultivo, concentración espermática, la presencia del "cumulus oophorus" y el volumen de medio de cocultivo utilizados. / In the present work, we have investigated the influence of differen factors, related to porcine gametes coculture, on the results of "in vitro" fertilization (IVF). We have try to improve the efficiency of the current system to get viable embryos (monospermic fertilizations). 129 prepuberal gilts have been used after the induction of ovulation by administration of 1250 I.U. of PMSG followed, 55 hours later, by 750 I.U. of HCG. The results showed that the best moment for the recovery of oocytes was 44 h after HCC administration. In the same way, the treatment followed was effective for the required objectives in 80.62% of the studied females and the medium number of recovered oocytes was 19.07 + 1.52 per animal, giving a total number 01 1984 oocytes used. In relation with the conditions of the fertilization system, the first investigated factor was the coculture time, understanding it as contact time between gametes. To study the effect of this factor, two experiences were realized; fot the first one, 4, 6 or 8 hours of coculture time were used (experience lb) The best results were obtained at 4 hours of coculture, because the percentage of penetration was maintained high (82.68%) and, at the same time, the percentage of monospermy increased (p<O.Ol), although the sperm concentration employed was deliberately high (12 x lOS spx/ml). The second investigated factor was the sperm concentration. The results showed that, among the used concentrations (3, 6 and 12 x lO 5 alive spz/ml), the maximum effectiveness in our system was obtained for the concntration of 6 x lOS spz/ml, since the percentage of penetration was not signficatively different of that obtained with the highest sperm concentration The third studied factor was the influence of the presence or absence of the expanded ."cumulus", which is shed with the oocytes at the ovulation, in the coculture medium. The presence of these cells joined with the intercellular matrix of hialuronic acid was highly beneficious for the improvement of the IVF, because of the percentage of penetration with the denuded oocytes (53.69%) was lower (p<0.01) than that obtained with the .cumulus" enclosed oocytes (69.16%), and the percentage of monospermy was higher (p<0.01) at the second case (39.45% vs. 60.97%). Finally, the percentage of penetration and monspermy was investigated using two different coculture medium volume, one the commonly used by other authors (2 ml) and another minor volume (0.4 mI). The results showed that the percentage of monospermy was higher (p<0.01) with the 0.4 mI volume (57.53 vs.78312%). We may deduce from the total results that the percentages of and monospermy in porcine IVF are due to the influence of different factors, some of them being the coculture time, the sperm concentrarían, the presence of "cumulus". and the coculture medium volume.

Moderne Schlachtsysteme tragen ein Risiko lebende Tiere weiterzuverarbeiten (SCHÜTTE und BOSTELMANN 2001, TROEGER 2005 und TROEGER und MEILER 2006). Gründe für dieses ernst zu nehmende tierschutzrelevante Problem sind eine ineffiziente Betäubung und/oder der Mangel an einer ausreichenden und schnellen Entblutung der Schweine. Die europäische Kommission hat 2009 die Verordnung (EG) Nr. 1099/2009 über den Schutz von Tieren zum Zeitpunkt der Tötung implementiert (ANON. 2009). Die Studie schafft Grundlagen um eine automatisierte Methode zu entwickeln, welche die Abwesenheit von Lebenszeichen von Schlachtschweinen verifiziert. Die Hypothese hierbei ist die Annahme, dass Schweine, die auf einen schmerzhaften Reiz wie heißes Wasser mehrere Minuten nach der Entblutung reagieren mit dem Risiko eines zumindest teilweise funktionierenden Gehirns behaftet sind. Die Studie fand an drei kommerziellen Schlachthöfen in Deutschland statt, die verschiedene elektrische Betäubungs- (Kopf-zu-Herz-Durchströmung) und Stechverfahren verwendeten. Insgesamt wurden am Schlachtband 5.301 Mastschweine im Hauptversuch untersucht und mittels Videokameras aufgezeichnet. Um die Abwesenheit von Lebenszeichen am Ende der Nachentblutestrecke, das heißt vier bis fünf Minuten nach Entblutestich, zu überprüfen wurde ein Heißwasser-Reiz mit 65 °C verwendet. Die Dauer der Reizapplikation betrug fünf bzw. 15 Sekunden. Eine automatisierte Reizapplikations-Anlage, erbaut von der Firma BANSS Schlacht- und Fördertechnik GmbH (Biedenkopf), induzierte den Stimulus vor allem im Bereich des Gesichts der Schweine. Als Referenz zu den Beobachtungen der Bewegungen während der Reiz-Applikation wurden Gehirnnerven-Reflexe (Corneal- und Lidschlussreflexe) und Reaktionen auf einen Kniff in die Nasenscheidewand klinisch untersucht. Schweine mit positiven Befunden wurden mittels Bolzenschuss nachbetäubt bzw. getötet. Die Sensitivität des Heißwasser-Tests lag bei 99 %. Eines von 75 Tieren wies positive Corneal- und Lidschlussreflexe auf, obwohl dieses Schwein auf den Heißwasser-Reiz nicht reagiert hatte. Jedoch konnten deutlich erkennbare Spontanbewegungen jenes Tieres bereits vorher beobachtet werden. Die Spezifität des Heißwasser-Tests lag bei 98 %. Beinah jedes Schwein mit negativen Gehirnnerven-Befunden blieb während der Reizapplikation unauffällig. 3,8 % (n = 199) der untersuchten Schweine zeigten eine Reaktion auf den Heißwasser-Reiz. Es war kein Unterschied zwischen dem fünf bzw. 15 Sekunden anhaltendem Reiz zu ermitteln. Neben einer ineffizienten Entblutung kann die reversible Betäubung als ein weiterer möglicher Grund für dieses Ergebnis genannt werden. Die Elektrische Betäubung ist reversibel, solange kein Herzkammerflimmern sicher ausgelöst wird (HOENDERKEN et al. 1980 und VOGEL et al. 2010). Es kann geschlussfolgert werden, dass die elektrische Kopf-zu-Herz-Durchströmung, die in den hier dargestellten Schlachtbetrieben verwendet wurde, nicht immer zum Herzkammerflimmern geführt hatte. Die Anzahl der Reaktionen der Schweine auf das heiße Wasser war begrenzt. 92 % der Schweine, die den Kopf während der Reizapplikation geschüttelt hatten und 78 % derer, die eine aufrichtende Bewegung gezeigt hatten, wiesen mindestens einen positiven Gehirnnerven-Befund auf. Auffälligkeiten in den Vordergliedmaßen korrelierten zu 59 % und das Muster “Maul öffnen” zu 52 % mit positiven Gehirnnerven-Befunden. Bewegungen aus dem Becken bzw. den Hintergliedmaße heraus waren nur zu 21 % mit positiven Gehirnnerven-Befunden verbunden. Bei der Betrachtung der Bewegungsmusterkombinationen stellte die Autorin fest, dass nahezu keine Reaktion missachtet werden sollte. Lediglich das Muster „ausschließliche Bewegungen Becken/Hintergliedmaße“ korrelierte in keinem der 59 Fälle mit positiven Gehirnnerven-Befunden. Dieses Ergebnis deckt sich mit den Aussagen von GRANDIN (2013) und den Mitarbeitern des bsi Schwarzenbek (ANON. 2013a), die darauf hinwiesen, dass der caudale Körperabschnitt elektrisch betäubter Schlachtschweine zur Einschätzung ihres möglicherweise vorhandenen Bewusstseins keine Relevanz besitzt. Während der Untersuchungen wurden außerdem Spontanbewegungen der Schlachtschweine zwischen dem Stechen und dem Heißwasser-Test analysiert. Jedes Tier, das eine Reaktion auf die Heißwasser-Reiz-Applikation zeigte und mindestens einen positiven Befund in der Gehirnnerven-Untersuchung aufwies, hatte vorher das Spontanbewegungsmuster „Maul öffnen“ durchgeführt. Um dem Überwachungs- und Schlachthofpersonal zu vermitteln, welche Spontan-bewegungsmuster bzw. reizinduzierten Reaktionen bei der Beobachtung der Nachentblutestrecke entscheidend sind, wurden entsprechende Arbeitsanweisungen entwickelt. Bereits vorhandene Literatur wurde hierbei mit eingearbeitet (ANIL 1991, ATKINSON et al. 2012 und EFSA 2013). Als eine wirksame Lösung um den sicheren Tod von Schweinen vor weiteren Schlachtarbeiten sicherzustellen, scheint es sinnvoll entsprechende Spontanbewegungen zu beachten und die Implementierung einer abschließenden Untersuchung mittels eines Heißwasser-Reiz-Tests am Ende der Nachentblutestrecke zu verwirklichen. Selbstverständlich sollten weiterhin die Betäubung und Entblutung der Tiere möglichst sicher kontrolliert werden. Für die Nachbetäubung bzw. Tötung der betroffenen Schweine wird der Einsatz eines Bolzenschussgerätes von der EFSA (2004), dem bsi Schwarzenbek und dem Max Rubner-Institut empfohlen. / Modern slaughter regimes carry a risk of live animals being further processed (SCHÜTTE and BOSTELMANN 2001, TROEGER 2005 and TROEGER and MEILER 2006). This serious animal welfare problem may result from inefficient stunning and/or lack of complete and fast exsanguination of the pigs. In 2009, the European Commission implemented Council Regulation (EC) No. 1099/2009 on the protection of animals at the time of killing (ANON. 2009). The study lays groundwork for developing an automated method to verify the absence of signs of life in slaughter pigs. The hypothesis is that pigs that react to a painful stimulus, like hot water, several minutes after debleeding have the risk of a partly functional brains. The study took place at three commercial abattoirs in Germany using different electrical stunning (head-to-body) and bleeding methods. In the main part of the study a total of 5,301 finishing pigs was examined and videotaped on line. As a stimulus to check the absence of signs of life right before further processing, namely four to five minutes after sticking, a hot-water-stimulus at 65 °C was utilized. The residence time of the stimulus amounted either five or 15 seconds. An automated construction, built by the company BANSS Schlacht- und Fördertechnik GmbH (Biedenkopf/Germany), implemented the stimulus mainly within the faces of the pigs. As a reference to the observation of movements during the stimulation, brain stem reflexes (corneal and palpebral) and reactions to a nasal septum pinch were clinically examined. Pigs with any positive result were restunned or killed using a captive bolt device. The sensitivity of the hot-water-test was determined at 99 %. One out of 75 animals exhibited positive corneal- and palpebral-reflexes although this one pig did not show any reaction to the hot water stimulation. However, obvious spontaneous movements of this animal could be observed beforehand. The specificity of the hot-water-test was determined at 98 %. Almost every pig with negative brain stem results remained motionless during the stimulation. A share of 3.8 % (n = 199) of pigs showed movements during the hot water exposure. Es war kein Unterschied zwischen dem fünf bzw. 15 Sekunden anhaltendem Reiz zu ermitteln. No difference was estimated between the residence times of five versus 15 seconds. Besides inefficient bleeding one possible reason for this result is reversible stunning. Electrical stunning is reversible, unless effective cardiac arrest is caused (HOENDERKEN et al. 1980 and VOGEL et al. 2010). It may be assumed that after head-to-body electrical stunning used by the abattoirs displayed in this study cardiac arrest was not always achieved. The number of individual responses was limited. 92 % of pigs that shook their heads during the stimulation and 78 % that showed a righting reflex exhibited at least one positive brain stem result. Noticeable front leg activity correlated to 59 % and the movement “opening of the mouth” to 52 % with positive brain stem results. Hips or hind leg movements were only associated with positive brain stem results in 21 % of the cases. By looking at the combinations of movements the author found that nearly no reaction should be ignored. Merely exclusive hips or hind leg movements in none of the 59 cases correlated with brain stem results. This finding is supported by the statements made by GRANDIN (2013) and the staff of the bsi Schwarzenbek (ANON. 2013a), pointing out that the caudal body part of electrically stunned slaughter pigs possesses no relevance to evaluating possible consciousness. During the study additionally spontaneous movements of the slaughter pigs were analyzed between sticking and the hot water device. Every animal that eventually showed a reaction to the hot water stimulation and exhibited at least one positive result during the brain stem examination had shown spontaneous mouth opening. Appropriate working instructions for the monitoring personnel and the slaughter staff, in order for them to realize which spontaneous movements or stimulus induced reactions during the observations of the debleeding line are relevant, were designed. For this available expertise has additionally been taken into account (ANIL 1991, ATKINSON et al. 2012 and EFSA 2013). As a suitable solution for ascertaining death before further processing, the idea of paying attention to slaughter pigs that obviously show signs of recovery and the implementation of a “last check” by using a hot water test right before further processing seems reasonable. Of course the stunning and exsanguination should still be safely monitored. The use of a captive bolt device to restun or kill “suspicious” pigs is recommended by the EFSA (2004), the bsi Schwarzenbek and the Max Rubner-Institute.

No description available.

La capacit?? de r??g??n??ration des foies de mammif??res est consid??rable. Suite ?? une h??patectomie partielle, les h??patocytes entrent de fa??on synchrone dans un ??tat de prolif??ration dans lequel ils quittent simultan??ment la phase G0 pour entrer en phase G1. Nous nous sommes int??ress??s aux changements ??pig??n??tiques et transcriptionnels qui surviennent durant ce processus. Pour ce faire, nous avons g??n??r?? des donn??es de ChIP-seq pour diff??rents temps suivant cette chirurgie pour : - la polym??rase II (pol II), - la polym??rase III (pol III), - les marqueurs ??pig??n??tiques H3K4me3 et H3K36me3, - ainsi que le cofacteur de transcription HCF-1. Ces donn??es nous ont permis de suivre et d???explorer les modifications dans la transcription et les changements ?? travers le g??nome durant la r??g??n??ration h??patique. La chromatine a ??t?? pr??par??e ?? partir de huit points dans le temps diff??rents suivant l'h??patectomie partielle (1h, 10h, 20h, 28h, 36h, 44h, 48h et 60h). Les ??chantillons ont ??t?? s??quenc??s avec la technologie dite "paired-end" (chaque fragment donne lieu ?? deux lectures de 50 ou 100 nucl??otides, une pour chaque bout) permettant la localisation pr??cise et la longueur exacte de chaque fragment s??quenc?? sur le g??nome. En moyenne, 225 millions de s??quences ont ??t?? obtenues pour chaque ??chantillon, puis cartographi??es sur le g??nome murin (C57BL6/J, assemblage MGSCv37/mm9, juillet 2007). De fa??on consistante avec une synchronisation robuste du cycle de division cellulaire, les patrons de transcription des g??nes du cycle cellulaire (par exemple, les cdks et les cyclines) r??v??lent un ??tat actif ou inactif bien d??fini qui corr??le avec l'activit?? attendue du g??ne. Nous avons effectu?? une premi??re analyse globale de l'occupation par la pol II ?? travers les diff??rents points dans le temps et avons identifi?? 9 423 g??nes montrant un changement significatif dans la fixation de la polym??rase au promoteur. En groupant les g??nes ayant un profil similaire, nous avons remarqu?? une concordance significative entre le point dans le temps du cycle cellulaire pendant lequel la pol II est pr??sente de fa??on maximale au promoteur des g??nes et la fonction qui leur est reli??e. Nous continuons d???explorer g??ne par g??ne, les variations au niveau des marqueurs ??pig??n??tiques, en fonction du profil de fixation de la pol II. Aucune analyse n???a cependant encore d??but?? en ce qui concerne la pol III et le facteur de transcription HCF-1. L'analyse continue de la transcription des g??nes du cycle de division cellulaire durant ce processus de r??g??n??ration h??patique vous sera pr??sent??e.

The accumulation of androstenone in the fat of mature boars results in boar taint; the conjugation of androstenone would decrease this important meat quality problem by decreasing the accumulation and increasing the excretion of androstenone. Leydig cells and testis microsomes from mature boars were incubated with radiolabeled pregnenolone, and the free and conjugated metabolites were examined by HPLC. Sulfated androstenone with a mass of 367 m/z was directly identified by MS, with a novel tentative structure of 3-keto-4- sulfoxy-androstenone. Addition of enolase to the microsomal incubations increased the formation of 3-keto-4-sulfoxy-androstenone. Overexpression of SULT2A1 in HEK cells resulted in the sulfoconjugation of dehydroepiandrosterone, but not androstenone, suggesting that SULT2A1 may not be involved in sulfoconjugation of androstenone. This thesis describes the novel direct characterization of androstenone sulfate and the importance of enolase in its formation. The relevance to boar taint metabolism is discussed.

Bone discolouration of pig carcasses is a quality concern that has been observed in Ontario slaughter plants. The objectives of this study were to establish the prevalence of pig carcasses showing bone discolouration, its relationship with residues of tetracyclines in bones, and to investigate the use of tetracyclines in feeding programs for grower-finisher pigs as the main risk factor for discolouration. Abattoir data were examined to determine the extent of the problem and the prevalence of bone discolouration during 2006, 2008, 2009, and 2010 was found to be 0.13%, 0.22%, 0.26%, and 0.28%, respectively, indicating that the issue of bone discolouration was present at low levels over the entire period of the study. A controlled trial using feed, water, and injectable tetracycline products to investigate the effect of tetracyclines on residue and bone colour was conducted. Bones were assessed visually for signs of discolouration, and high performance liquid chromatography (HPLC) was used to measure the levels of tetracycline residues in the bones. Results from this trial demonstrated that discolouration could be produced with 660ppm of chlortetracycline (CTC) in feed for 12 weeks even when 33 days of withdrawal time was observed. It was also found that residues of tetracyclines can be present in bones in the absence of discolouration. A retrospective study was conducted to investigate tetracycline use in herds identified as having discoloured bones at slaughter. Positive shipments were associated with dosage and duration of CTC use as well as with length of withdrawal. In conclusion, discoloured bones of pig carcasses were identified at low levels in one large Ontario abattoir; however, further investigation is needed in order to determine the impact it may have on the swine industry. / Ontario Pork and the University of Guelph - Ontario Ministry of Agriculture (OMAFRA) Sustainable Production System Program

Terminal restriction fragment length polymorphism (T-RFLP) analysis and pyrosequencing were used to characterize the microbiota of the tonsil of the soft palate of 126 unfit and 18 healthy pigs. The T-RFLP analysis method was first optimized for the study of the pig tonsil microbiota and the data compared with culture-based identification of common pig pathogens. Putative identifications of the members of the microbiota revealed that the phyla Firmicutes, Proteobacteria and Bacteroidetes were the most prevalent. A comparison of the T-RFLP analysis results grouped into clusters to clinical conditions revealed paleness, abscess, PRRS virus, and Mycoplasma hyopneumoniae to be significantly associated with cluster membership. T-RFLP analysis was also used to select representative tonsil samples for pyrosequencing. These studies confirmed Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, and Proteobacteria to be the core phyla of the microbiota of the tonsil of the soft palate of pigs. / OMAFRA Animal Health Strategic Investment