Dissertations / Theses: 'FISIOLOGIA VETERINARIA'

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Bovo, Rafael Parelli [UNESP]. "Fisiologia térmica e balanço hídrico em anfíbios anuros." Universidade Estadual Paulista (UNESP), 2015. http://hdl.handle.net/11449/134132.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Temperatura e disponibilidade de água são fatores que afetam importantemente a ecologia, a fisiologia e o comportamento de anfíbios. Nesse sentido, estes animais necessitam apresentar uma série de características ecofisiológicas que permita a eles lidarem potencialmente com aspectos ambientais desafiadores. Na presente tese, objetivei explorar diferentes tópicos (gradientes ecológicos, doenças e atividade) enfocando a fisiologia térmica e o balanço hídrico de anfíbios. O primeiro capítulo apresenta uma introdução geral abordando um dos temas centrais da presente tese, a fisiologia térmica de anfíbios. Entretanto, este capítulo também explora a inter-relação entre regulação da temperatura corpórea e balanço de água. O segundo capítulo investiga se parâmetros fisiológicos associados à tolerância térmica e ao balanço hídrico, em cinco espécies de anfíbios anuros, variam ao longo de gradientes altitudinais em duas cadeias montanhosas, Serra do Mar e Serra da Mantiqueira, da Floresta Atlântica. O terceiro capítulo examina os potenciais efeitos da quitridiomicose - uma doença emergente causada por fungo, considerada responsável por declínios populacionais e extinções de anfíbios em todo o mundo devido a alterações osmorregulatórias cutâneas - sobre o balanço hídrico e mortalidade de espécies de anfíbios anuros do Brasil. O quarto capítulo investiga se a atividade de dispersão diurna em juvenis do sapo Dermatonotus muelleri estaria correlacionada com parâmetros fisiológicos associados à fisiologia térmica e balanço hídrico
Temperature and water availability are factors that affect, importantly, the ecology, physiology, and behavior of amphibians. In this sense, these animals need to present a number of ecophysiological characteristics that allows them to cope with potentially challenging environmental features. In this thesis, I aimed to explore different topics (ecological gradients, diseases, and activity) focusing on the thermal physiology and water balance of anuran amphibians. The first chapter presents a general introduction focused on one of the central themes of this thesis, the thermal physiology of amphibians. However, this chapter also explores the inter-relationship between body temperature regulation and water balance. The second chapter investigates whether or not physiological traits associated to thermal tolerances and water balance, in five anuran amphibians, vary along elevational gradients along two mountain ranges, Serra do Mar and Serra da Mantiqueira, in the Brazilian Atlantic Forest. The third chapter examines the potential effects of chytridomycosis - an emerging fungal disease claimed to cause amphibian population declines and extinctions worldwide due to cutaneous osmoregulatory disorders - on the water balance and mortality of anuran species from Brazil. The fourth chapter investigates whether or not the diurnal activity of dispersing juveniles of the burrowing frog, Dermatonotus muelleri, might be correlated with physiological traits associated to thermal physiology and water balance
FAPESP: 10/20061-6

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2

Santini, Sujen Eleonora <1979&gt. "Il 2-metossiestradiolo: un potenziale inibitore fisiologico dell'angiogenesi follicolare." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2008. http://amsdottorato.unibo.it/809/1/Tesi_Santini_Sujen_Eleonora.pdf.

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3

Santini, Sujen Eleonora <1979&gt. "Il 2-metossiestradiolo: un potenziale inibitore fisiologico dell'angiogenesi follicolare." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2008. http://amsdottorato.unibo.it/809/.

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4

Turba, Maria Elena <1978&gt. "Studio, mediante applicazioni biotecnologiche, di tre diversi modelli spontanei o indotti di patologie del sistema nervoso." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2007. http://amsdottorato.unibo.it/31/1/Tesi_Dottorato_Turba_Maria_Elena.pdf.

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5

Turba, Maria Elena <1978&gt. "Studio, mediante applicazioni biotecnologiche, di tre diversi modelli spontanei o indotti di patologie del sistema nervoso." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2007. http://amsdottorato.unibo.it/31/.

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6

De, Andrea Ribeiro Luciana <1971&gt. "Angiogenesis and angioregression gene expression analyses in swine corpus luteum." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2007. http://amsdottorato.unibo.it/428/1/Tesi_dottorato_Luciana_de_Andrea_Ribeiro_-_XIX_ciclo.pdf.

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The corpus luteum (CL) lifespan is characterized by a rapid growth, differentiation and controlled regression of the luteal tissue, accompanied by an intense angiogenesis and angioregression. Indeed, the CL is one of the most highly vascularised tissue in the body with a proliferation rate of the endothelial cells 4- to 20-fold more intense than in some of the most malignant human tumours. This angiogenic process should be rigorously controlled to allow the repeated opportunities of fertilization. After a first period of rapid growth, the tissue becomes stably organized and prepares itself to switch to the phenotype required for its next apoptotic regression. In pregnant swine, the lifespan of the CLs must be extended to support embryonic and foetal development and vascularisation is necessary for the maintenance of luteal function. Among the molecules involved in the angiogenesis, Vascular Endothelial Growth Factor (VEGF) is the main regulator, promoting endothelial cells proliferation, differentiation and survival as well as vascular permeability and vessel lumen formation. During vascular invasion and apoptosis process, the remodelling of the extracellular matrix is essential for the correct evolution of the CL, particularly by the action of specific class of proteolytic enzymes known as matrix metalloproteinases (MMPs). Another important factor that plays a role in the processes of angiogenesis and angioregression during the CL formation and luteolysis is the isopeptide Endothelin-1 (ET-1), which is well-known to be a potent vasoconstrictor and mitogen for endothelial cells. The goal of the present thesis was to study the role and regulation of vascularisation in an adult vascular bed. For this purpose, using a precisely controlled in vivo model of swine CL development and regression, we determined the levels of expression of the members of VEGF system (VEGF total and specific isoforms; VEGF receptor-1, VEGFR-1; VEGF receptor-2, VEGFR-2) and ET- 1 system (ET-1; endothelin converting enzyme-1, ECE-1; endothelin receptor type A, ET-A) as well as the activity of the Ca++/Mg++-dependent endonucleases and gelatinases (MMP-2 and MMP-9). Three experiments were conducted to reach such objectives in CLs isolated from ovaries of cyclic, pregnant or fasted gilts. In the Experiment I, we evaluated the influence of acute fasting on VEGF production and VEGF, VEGFR-2, ET-1, ECE-1 and ET-A mRNA expressions in CLs collected on day 6 after ovulation (midluteal phase). The results indicated a down-regulation of VEGF, VEGFR-2, ET-1 and ECE-1 mRNA expression, although no change was observed for VEGF protein. Furthermore, we observed that fasting stimulated steroidogenesis by luteal cells. On the basis of the main effects of VEGF (stimulation of vessel growth and endothelial permeability) and ET-1 (stimulation of endothelial cell proliferation and vasoconstriction, as well as VEGF stimulation), we concluded that feed restriction possibly inhibited luteal vessel development. This could be, at least in part, compensated by a decrease of vasal tone due to a diminution of ET-1, thus ensuring an adequate blood flow and the production of steroids by the luteal cells. In the Experiment II, we investigated the relationship between VEGF, gelatinases and Ca++/Mg++-dependent endonucleases activities with the functional CL stage throughout the oestrous cycle and at pregnancy. The results demonstrated differential patterns of expression of those molecules in correspondence to the different phases of the oestrous cycle. Immediately after ovulation, VEGF mRNA/protein levels and MMP-9 activity are maximal. On days 5–14 after ovulation, VEGF expression and MMP-2 and -9 activities are at basal levels, while Ca++/Mg++-dependent endonuclease levels increased significantly in relation to day 1. Only at luteolysis (day 17), Ca++/Mg++-dependent endonuclease and MMP-2 spontaneous activity increased significantly. At pregnancy, high levels of MMP-9 and VEGF were observed. These results suggested that during the very early luteal phase, high MMPs activities coupled with high VEGF levels drive the tissue to an angiogenic phenotype, allowing CL growth under LH (Luteinising Hormone) stimulus, while during the late luteal phase, low VEGF and elevate MMPs levels may play a role in the apoptotic tissue and extracellular matrix remodelling during structural luteolysis. In the Experiment III, we described the expression patterns of all distinct VEGF isoforms throughout the oestrous cycle. Furthermore, the mRNA expression and protein levels of both VEGF receptors were also evaluated. Four novel VEGF isoforms (VEGF144, VEGF147, VEGF182, and VEGF164b) were found for the first time in swine and the seven identified isoforms presented four different patterns of expression. All isoforms showed their highest mRNA levels in newly formed CLs (day 1), followed by a decrease during mid-late luteal phase (days 10–17), except for VEGF182, VEGF188 and VEGF144 that showed a differential regulation during late luteal phase (day 14) or at luteolysis (day 17). VEGF protein levels paralleled the most expressed and secreted VEGF120 and VEGF164 isoforms. The VEGF receptors mRNAs showed a different pattern of expression in relation to their ligands, increasing between day 1 and 3 and gradually decreasing during the mid-late luteal phase. The differential regulation of some VEGF isoforms principally during the late luteal phase and luteolysis suggested a specific role of VEGF during tissue remodelling process that occurs either for CL maintenance in case of pregnancy or for noncapillary vessel development essential for tissue removal during structural luteolysis. In summary, our findings allow us to determine relationships among factors involved in the angiogenesis and angioregression mechanisms that take place during the formation and regression of the CL. Thus, CL provides a very interesting model for studying such factors in different fields of the basic research.

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7

De, Andrea Ribeiro Luciana <1971&gt. "Angiogenesis and angioregression gene expression analyses in swine corpus luteum." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2007. http://amsdottorato.unibo.it/428/.

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Abstract:

The corpus luteum (CL) lifespan is characterized by a rapid growth, differentiation and controlled regression of the luteal tissue, accompanied by an intense angiogenesis and angioregression. Indeed, the CL is one of the most highly vascularised tissue in the body with a proliferation rate of the endothelial cells 4- to 20-fold more intense than in some of the most malignant human tumours. This angiogenic process should be rigorously controlled to allow the repeated opportunities of fertilization. After a first period of rapid growth, the tissue becomes stably organized and prepares itself to switch to the phenotype required for its next apoptotic regression. In pregnant swine, the lifespan of the CLs must be extended to support embryonic and foetal development and vascularisation is necessary for the maintenance of luteal function. Among the molecules involved in the angiogenesis, Vascular Endothelial Growth Factor (VEGF) is the main regulator, promoting endothelial cells proliferation, differentiation and survival as well as vascular permeability and vessel lumen formation. During vascular invasion and apoptosis process, the remodelling of the extracellular matrix is essential for the correct evolution of the CL, particularly by the action of specific class of proteolytic enzymes known as matrix metalloproteinases (MMPs). Another important factor that plays a role in the processes of angiogenesis and angioregression during the CL formation and luteolysis is the isopeptide Endothelin-1 (ET-1), which is well-known to be a potent vasoconstrictor and mitogen for endothelial cells. The goal of the present thesis was to study the role and regulation of vascularisation in an adult vascular bed. For this purpose, using a precisely controlled in vivo model of swine CL development and regression, we determined the levels of expression of the members of VEGF system (VEGF total and specific isoforms; VEGF receptor-1, VEGFR-1; VEGF receptor-2, VEGFR-2) and ET- 1 system (ET-1; endothelin converting enzyme-1, ECE-1; endothelin receptor type A, ET-A) as well as the activity of the Ca++/Mg++-dependent endonucleases and gelatinases (MMP-2 and MMP-9). Three experiments were conducted to reach such objectives in CLs isolated from ovaries of cyclic, pregnant or fasted gilts. In the Experiment I, we evaluated the influence of acute fasting on VEGF production and VEGF, VEGFR-2, ET-1, ECE-1 and ET-A mRNA expressions in CLs collected on day 6 after ovulation (midluteal phase). The results indicated a down-regulation of VEGF, VEGFR-2, ET-1 and ECE-1 mRNA expression, although no change was observed for VEGF protein. Furthermore, we observed that fasting stimulated steroidogenesis by luteal cells. On the basis of the main effects of VEGF (stimulation of vessel growth and endothelial permeability) and ET-1 (stimulation of endothelial cell proliferation and vasoconstriction, as well as VEGF stimulation), we concluded that feed restriction possibly inhibited luteal vessel development. This could be, at least in part, compensated by a decrease of vasal tone due to a diminution of ET-1, thus ensuring an adequate blood flow and the production of steroids by the luteal cells. In the Experiment II, we investigated the relationship between VEGF, gelatinases and Ca++/Mg++-dependent endonucleases activities with the functional CL stage throughout the oestrous cycle and at pregnancy. The results demonstrated differential patterns of expression of those molecules in correspondence to the different phases of the oestrous cycle. Immediately after ovulation, VEGF mRNA/protein levels and MMP-9 activity are maximal. On days 5–14 after ovulation, VEGF expression and MMP-2 and -9 activities are at basal levels, while Ca++/Mg++-dependent endonuclease levels increased significantly in relation to day 1. Only at luteolysis (day 17), Ca++/Mg++-dependent endonuclease and MMP-2 spontaneous activity increased significantly. At pregnancy, high levels of MMP-9 and VEGF were observed. These results suggested that during the very early luteal phase, high MMPs activities coupled with high VEGF levels drive the tissue to an angiogenic phenotype, allowing CL growth under LH (Luteinising Hormone) stimulus, while during the late luteal phase, low VEGF and elevate MMPs levels may play a role in the apoptotic tissue and extracellular matrix remodelling during structural luteolysis. In the Experiment III, we described the expression patterns of all distinct VEGF isoforms throughout the oestrous cycle. Furthermore, the mRNA expression and protein levels of both VEGF receptors were also evaluated. Four novel VEGF isoforms (VEGF144, VEGF147, VEGF182, and VEGF164b) were found for the first time in swine and the seven identified isoforms presented four different patterns of expression. All isoforms showed their highest mRNA levels in newly formed CLs (day 1), followed by a decrease during mid-late luteal phase (days 10–17), except for VEGF182, VEGF188 and VEGF144 that showed a differential regulation during late luteal phase (day 14) or at luteolysis (day 17). VEGF protein levels paralleled the most expressed and secreted VEGF120 and VEGF164 isoforms. The VEGF receptors mRNAs showed a different pattern of expression in relation to their ligands, increasing between day 1 and 3 and gradually decreasing during the mid-late luteal phase. The differential regulation of some VEGF isoforms principally during the late luteal phase and luteolysis suggested a specific role of VEGF during tissue remodelling process that occurs either for CL maintenance in case of pregnancy or for noncapillary vessel development essential for tissue removal during structural luteolysis. In summary, our findings allow us to determine relationships among factors involved in the angiogenesis and angioregression mechanisms that take place during the formation and regression of the CL. Thus, CL provides a very interesting model for studying such factors in different fields of the basic research.

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8

Viggiani, Roberta <1974&gt. "La determinazione del cortisolo nel pelo per la valutazione del benessere animale." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2008. http://amsdottorato.unibo.it/810/1/Tesi_Viggiani_Roberta.pdf.

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La definizione di “benessere animale” e le modalità di determinazione di tale parametro sono ancora ampiamente dibattute. C’è, però, una generale concordanza sul fatto che una condizione di malessere dia origine a variazioni fisiologiche e comportamentali che possono essere rilevate e misurate. Tra i parametri endocrini, il più studiato è, senza dubbio, il cortisolo, in quanto connesso con l’attivazione dell’asse ipotalamico-pituitario-surrenale in condizioni di stress e quindi ritenuto indicatore ideale di benessere, benché debba essere utilizzato con cautela in quanto un aumento dei livelli di questo ormone non si verifica con ogni tipo di stressor. Inoltre, si deve considerare che la raccolta del campione per effettuare le analisi, spesso implica il confinamento ed il contenimento degli animali e può essere, quindi, essa stessa un fattore stressante andando ad alterare i risultati. Alla luce delle suddette conoscenze gli obiettivi scientifici di questa ricerca, condotta sul gatto e sul cane, sono stati innanzitutto validare il metodo di dosaggio di cortisolo dal pelo e stabilire se tale dosaggio può rappresentare un indicatore, non invasivo, di benessere dell’animale (indice di “stress cronico”). In seguito, abbiamo voluto individuare i fattori di stress psico-sociale in gatti che vivono in gattile, in condizioni di alta densità, analizzando i correlati comportamentali ed ormonali dello stress e del benessere in questa condizione socio-ecologica, ricercando, in particolare, l’evidenza ormonale di uno stato di stress prolungato e la messa in atto di strategie comportamentali di contenimento dello stesso e il ruolo della marcatura visivo-feromonale, inoltre abbiamo effettuato un confronto tra oasi feline di diversa estensione spaziale per valutare come varia lo stress in rapporto allo spazio disponibile. Invece, nel cane abbiamo voluto evidenziare eventuali differenze dei livelli ormonali tra cani di proprietà e cani di canili, tra cani ospitati in diversi canili e tra cani che vivono in diverse realtà familiari; abbiamo voluto valutare gli effetti di alcuni arricchimenti sui cani di canile ed, infine, abbiamo analizzato cani sottoposti a specifici programmi si addestramento. Il primo importante ed originale risultato raggiunto, che risponde al primo obiettivo della ricerca, è stato la validazione del dosaggio radioimmunologico di cortisolo in campioni di pelo. Questo risultato, a nostro avviso, apre una nuova finestra sul campo della diagnostica endocrinologica metabolica. Attualmente, infatti, il monitoraggio ormonale viene effettuato su campioni ematici la cui raccolta prevede un elevato stress (stress da prelievo) per l’animale data l'invasività dell'operazione che modifica l’attività di ipotalamo-ipofisi-surrene e, dunque, provoca repentine alterazioni delle concentrazioni ormonali. Questa metodica offre, quindi, il vantaggio dell’estrema semplicità di raccolta del campione e, in più, il bassissimo costo del materiale utilizzato. Dalle ricerche condotte sui gatti di gattile sono scaturite preziose indicazioni per future indagini sullo stress e sul comportamento sociale felino. I risultati dell’analisi congiunta del comportamento e delle concentrazioni ormonali hanno evidenziato che la disponibilità di postazioni di marcatura visivo-feromonale ha un effetto positivo sia sugli indicatori comportamentali, sia su quelli ormonali di stress. I risultati dell’analisi delle concentrazioni di cortisolo, derivanti dal confronto tra sette oasi feline di diversa estensione spaziale hanno permesso di evidenziare un aumento dei livelli dell’ormone inversamente proporzionale allo spazio disponibile. Lo spazio disponibile, però, non è l’unico fattore da prendere in considerazione al fine di assicurare il benessere dell’animale infatti, nelle colonie che presentavano instabilità sociale e variabilità territoriale il cortisolo aveva valori elevati nonostante le notevoli disponibilità di spazio. Infine, si è potuto costatare come anche lo stare appartati, aumenti proporzionalmente con l’aumentare dello spazio. Questo comportamento risulta essere molto importante in quanto mitiga lo stress ed è da prendere in considerazione nell’allestimento di colonie feline. Infatti, nelle colonie di dimensioni ridotte dove lo stress è già alto, l’impossibilità dei soggetti di appartarsi può contribuire a peggiorare la situazione; ecco perché si dovrebbero creare luoghi artificiali per fornire ai gatti la possibilità di appartarsi, magari sfruttando gli spazi sopraelevati (tetti, alberi, ecc.). Per quanto riguarda il confronto tra cani di proprietà e cani di canile non sono state evidenziate differenze significative nei livelli di cortisolo nel pelo mentre abbiamo rilevato che quest’ultimi sono influenzati dalla disponibilità di spazio: infatti sia i cani di proprietà che vivevano in giardino, sia i cani dei canili che praticavano lo sgambamento presentavano livelli di cortisolo nel pelo più bassi rispetto, rispettivamente, ai cani di proprietà che vivevano in appartamento o appartamento/giardino e a quelli di canile che non praticavano lo sgambamento. L’arricchimento ambientale fornito ai cani di canile ha esercitato un’influenza positiva riducendo i livelli di cortisolo e migliorando la docilità dei soggetti, favorendone un’eventuale adozione. Si è inoltre messo in luce che i programmi di addestramento, eseguiti con tecniche “gentili”, non comportano situazioni stressanti per l’animale e aiutano i cani ad esprimere doti di equilibrio che rimarrebbero altrimenti celate dagli aspetti più istintivi del carattere. D’altra parte, l’impegno agonistico prima di una competizione e il livello di addestramento raggiunto dai cani, influenzano le concentrazioni di cortisolo a riposo e durante l’esercizio fisico. Questi risultati possono sicuramente dare utili suggerimenti per la gestione e la cura di gatti e cani al fine di migliorarne le condizioni di benessere.

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9

Viggiani, Roberta <1974&gt. "La determinazione del cortisolo nel pelo per la valutazione del benessere animale." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2008. http://amsdottorato.unibo.it/810/.

Full text

Abstract:

La definizione di “benessere animale” e le modalità di determinazione di tale parametro sono ancora ampiamente dibattute. C’è, però, una generale concordanza sul fatto che una condizione di malessere dia origine a variazioni fisiologiche e comportamentali che possono essere rilevate e misurate. Tra i parametri endocrini, il più studiato è, senza dubbio, il cortisolo, in quanto connesso con l’attivazione dell’asse ipotalamico-pituitario-surrenale in condizioni di stress e quindi ritenuto indicatore ideale di benessere, benché debba essere utilizzato con cautela in quanto un aumento dei livelli di questo ormone non si verifica con ogni tipo di stressor. Inoltre, si deve considerare che la raccolta del campione per effettuare le analisi, spesso implica il confinamento ed il contenimento degli animali e può essere, quindi, essa stessa un fattore stressante andando ad alterare i risultati. Alla luce delle suddette conoscenze gli obiettivi scientifici di questa ricerca, condotta sul gatto e sul cane, sono stati innanzitutto validare il metodo di dosaggio di cortisolo dal pelo e stabilire se tale dosaggio può rappresentare un indicatore, non invasivo, di benessere dell’animale (indice di “stress cronico”). In seguito, abbiamo voluto individuare i fattori di stress psico-sociale in gatti che vivono in gattile, in condizioni di alta densità, analizzando i correlati comportamentali ed ormonali dello stress e del benessere in questa condizione socio-ecologica, ricercando, in particolare, l’evidenza ormonale di uno stato di stress prolungato e la messa in atto di strategie comportamentali di contenimento dello stesso e il ruolo della marcatura visivo-feromonale, inoltre abbiamo effettuato un confronto tra oasi feline di diversa estensione spaziale per valutare come varia lo stress in rapporto allo spazio disponibile. Invece, nel cane abbiamo voluto evidenziare eventuali differenze dei livelli ormonali tra cani di proprietà e cani di canili, tra cani ospitati in diversi canili e tra cani che vivono in diverse realtà familiari; abbiamo voluto valutare gli effetti di alcuni arricchimenti sui cani di canile ed, infine, abbiamo analizzato cani sottoposti a specifici programmi si addestramento. Il primo importante ed originale risultato raggiunto, che risponde al primo obiettivo della ricerca, è stato la validazione del dosaggio radioimmunologico di cortisolo in campioni di pelo. Questo risultato, a nostro avviso, apre una nuova finestra sul campo della diagnostica endocrinologica metabolica. Attualmente, infatti, il monitoraggio ormonale viene effettuato su campioni ematici la cui raccolta prevede un elevato stress (stress da prelievo) per l’animale data l'invasività dell'operazione che modifica l’attività di ipotalamo-ipofisi-surrene e, dunque, provoca repentine alterazioni delle concentrazioni ormonali. Questa metodica offre, quindi, il vantaggio dell’estrema semplicità di raccolta del campione e, in più, il bassissimo costo del materiale utilizzato. Dalle ricerche condotte sui gatti di gattile sono scaturite preziose indicazioni per future indagini sullo stress e sul comportamento sociale felino. I risultati dell’analisi congiunta del comportamento e delle concentrazioni ormonali hanno evidenziato che la disponibilità di postazioni di marcatura visivo-feromonale ha un effetto positivo sia sugli indicatori comportamentali, sia su quelli ormonali di stress. I risultati dell’analisi delle concentrazioni di cortisolo, derivanti dal confronto tra sette oasi feline di diversa estensione spaziale hanno permesso di evidenziare un aumento dei livelli dell’ormone inversamente proporzionale allo spazio disponibile. Lo spazio disponibile, però, non è l’unico fattore da prendere in considerazione al fine di assicurare il benessere dell’animale infatti, nelle colonie che presentavano instabilità sociale e variabilità territoriale il cortisolo aveva valori elevati nonostante le notevoli disponibilità di spazio. Infine, si è potuto costatare come anche lo stare appartati, aumenti proporzionalmente con l’aumentare dello spazio. Questo comportamento risulta essere molto importante in quanto mitiga lo stress ed è da prendere in considerazione nell’allestimento di colonie feline. Infatti, nelle colonie di dimensioni ridotte dove lo stress è già alto, l’impossibilità dei soggetti di appartarsi può contribuire a peggiorare la situazione; ecco perché si dovrebbero creare luoghi artificiali per fornire ai gatti la possibilità di appartarsi, magari sfruttando gli spazi sopraelevati (tetti, alberi, ecc.). Per quanto riguarda il confronto tra cani di proprietà e cani di canile non sono state evidenziate differenze significative nei livelli di cortisolo nel pelo mentre abbiamo rilevato che quest’ultimi sono influenzati dalla disponibilità di spazio: infatti sia i cani di proprietà che vivevano in giardino, sia i cani dei canili che praticavano lo sgambamento presentavano livelli di cortisolo nel pelo più bassi rispetto, rispettivamente, ai cani di proprietà che vivevano in appartamento o appartamento/giardino e a quelli di canile che non praticavano lo sgambamento. L’arricchimento ambientale fornito ai cani di canile ha esercitato un’influenza positiva riducendo i livelli di cortisolo e migliorando la docilità dei soggetti, favorendone un’eventuale adozione. Si è inoltre messo in luce che i programmi di addestramento, eseguiti con tecniche “gentili”, non comportano situazioni stressanti per l’animale e aiutano i cani ad esprimere doti di equilibrio che rimarrebbero altrimenti celate dagli aspetti più istintivi del carattere. D’altra parte, l’impegno agonistico prima di una competizione e il livello di addestramento raggiunto dai cani, influenzano le concentrazioni di cortisolo a riposo e durante l’esercizio fisico. Questi risultati possono sicuramente dare utili suggerimenti per la gestione e la cura di gatti e cani al fine di migliorarne le condizioni di benessere.

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10

Bucci, Diego <1981&gt. "Detection and localization of GLUTs in spermatozoa from different domestic species." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2353/1/Bucci_Diego_tesi.pdf.

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Abstract:

Sperm cells need hexoses as a substrate for their function, for both the maintenance of membrane homeostasis and the movement of the tail. These cells have a peculiar metabolism that has not yet been fully understood, but it is clear that they obtain energy from hexoses through glycolisis and/or oxidative phosphorylation. Spermatozoa are in contact with different external environments, beginning from the testicular and epididymal fluid, passing to the seminal plasma and finally to the female genital tract fluids; in addition, with the spread of reproductive biotechnologies, sperm cells are diluted and stored in various media, containing different energetic substrates. To utilize these energetic sources, sperm cells, as other eukaryotic cells, have a well-constructed protein system, that is mainly represented by the GLUT family proteins. These transporters have a membrane-spanning α-helix structure and work as an enzymatic pump that permit a fast gradient dependent passage of sugar molecules through the lipidic bilayer of sperm membrane. Many GLUTs have been studied in man, bull and rat spermatozoa; the presence of some GLUTs has been also demonstrated in boar and dog spermatozoa. The aims of the present study were - to determine the presence of GLUTs 1, 2, 3, 4 and 5 in boar, horse, dog and donkey spermatozoa and to describe their localization; - to study eventual changes in GLUTs location after capacitation and acrosome reaction in boar, stallion and dog spermatozoa; - to determine possible changes in GLUTs localization after capacitation induced by insulin and IGF stimulation in boar spermatozoa; - to evaluate changes in GLUTs localization after flow-cytometric sex sorting in boar sperm cells. GLUTs 1, 2, 3 and 5 presence and localization have been demonstrated in boar, stallion, dog and donkey spermatozoa by western blotting and immunofluorescence analysis; a relocation in GLUTs after capacitation has been observed only in dog sperm cells, while no changes have been observed in the other species examined. As for boar, the stimulation of the capacitation with insulin and IGF didn’t cause any change in GLUTs localization, as well as for the flow cytometric sorting procedure. In conclusion, this study confirms the presence of GLUTs 1, 2 ,3 and 5 in boar, dog, stallion and donkey spermatozoa, while GLUT 4 seems to be absent, as a confirmation of other studies. Only in dog sperm cells capacitating conditions induce a change in GLUTs distribution, even if the physiological role of these changes should be deepened.
Gli spermatozoi di mammifero necessitano di zuccheri per espletare le loro funzioni, come il mantenimento dell’omeostasi membranale ed il movimento. Queste cellule hanno un metabolismo particolare, che non è ancora stato del tutto compreso, anche se è chiaro che ottengano energia da esosi tramite il processo glicolitico e la fosforilazione ossidativa. Gli spermatozoi sono a contatto con ambienti esterni molto diversi: dai fluidi testicolare ed epididimale, per passare al plasma seminale ed infine alle secrezioni dell’apparato genitale femminile. Inoltre, con la diffusione delle biotecnologie riproduttive, il liquido seminale è diluito e conservato in svariati media contenenti diversi substrati energetici. Per sfruttare queste fonti energetiche gli spermatozoi, come le altre cellule eucariotiche, hanno un sistema proteico di membrana ben definito, rappresentato principalmente dalla famiglia dei GLUT. Queste proteine hanno una struttura transmembranale ad alfa elica e funzionano come una pompa enzimatica che permette un trasporto passivo veloce e secondo gradiente di concentrazione delle molecole di zucchero attraverso lo strato lipidico. Alcuni GLUT sono stati studiati negli spermatozoi di uomo, ratto e toro e la presenza di alcuni altri è stata dimostrata in cane e maiale. Gli scopi di questo studio sono stati: • determinare la presenza dei GLUT 1, 2, 3, 4 e 5 in spermatozoi di maiale, cane , stallone e asino e descrivere la loro localizzazione; • studiare eventuali cambi di localizzazione dovuti alla capacitazione o alla reazione acrosomiale in spermatozoi di maiale, cane e stallone; • valutare cambiamenti nella disposizione dei GLUT negli spermatozoi di maiale in seguito a capacitazione stimolata con insulina ed IGF; • valutare possibili cambiamenti di localizzazione dei GLUT a seguito del processo di “sessaggio” mediante citofluorimetro sorter. La presenza e la localizzazione dei GLUT 1, 2, 3 e 5 sono state dimostrate negli spermatozoi di maiale, asino, cavallo e cane mediante le tecniche di western blotting ed immunofluorescenza indiretta; una rilocalizzazione delle proteine dopo capacitazione è stata osservata solo negli spermatozoi di cane e nessun cambiamento è stato registrato nelle altre specie. Per quanto riguarda il maiale, non si sono rilevate rilocalizzazioni dei GLUT a seguito della capacitazione con stimolazione con insulina ed IGF e nemmeno a seguito del processo di “sex sorting”. Concludendo, questo studio conferma la presenza dei GLUT 1, 2, 3 e 5 negli spermatozoi di maiale, cane stallone e asino, mentre il GLUT 4 sembra essere assente, a conferma di alcuni studi precedenti. Solo negli spermatozoi di cane le condizioni capacitanti inducono un cambiamento nella distribuzione dei GLUT, anche se il ruolo fisiologico di questi cambiamenti deve essere ancora approfondito.

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11

Bucci, Diego <1981&gt. "Detection and localization of GLUTs in spermatozoa from different domestic species." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2353/.

Full text

Abstract:

Sperm cells need hexoses as a substrate for their function, for both the maintenance of membrane homeostasis and the movement of the tail. These cells have a peculiar metabolism that has not yet been fully understood, but it is clear that they obtain energy from hexoses through glycolisis and/or oxidative phosphorylation. Spermatozoa are in contact with different external environments, beginning from the testicular and epididymal fluid, passing to the seminal plasma and finally to the female genital tract fluids; in addition, with the spread of reproductive biotechnologies, sperm cells are diluted and stored in various media, containing different energetic substrates. To utilize these energetic sources, sperm cells, as other eukaryotic cells, have a well-constructed protein system, that is mainly represented by the GLUT family proteins. These transporters have a membrane-spanning α-helix structure and work as an enzymatic pump that permit a fast gradient dependent passage of sugar molecules through the lipidic bilayer of sperm membrane. Many GLUTs have been studied in man, bull and rat spermatozoa; the presence of some GLUTs has been also demonstrated in boar and dog spermatozoa. The aims of the present study were - to determine the presence of GLUTs 1, 2, 3, 4 and 5 in boar, horse, dog and donkey spermatozoa and to describe their localization; - to study eventual changes in GLUTs location after capacitation and acrosome reaction in boar, stallion and dog spermatozoa; - to determine possible changes in GLUTs localization after capacitation induced by insulin and IGF stimulation in boar spermatozoa; - to evaluate changes in GLUTs localization after flow-cytometric sex sorting in boar sperm cells. GLUTs 1, 2, 3 and 5 presence and localization have been demonstrated in boar, stallion, dog and donkey spermatozoa by western blotting and immunofluorescence analysis; a relocation in GLUTs after capacitation has been observed only in dog sperm cells, while no changes have been observed in the other species examined. As for boar, the stimulation of the capacitation with insulin and IGF didn’t cause any change in GLUTs localization, as well as for the flow cytometric sorting procedure. In conclusion, this study confirms the presence of GLUTs 1, 2 ,3 and 5 in boar, dog, stallion and donkey spermatozoa, while GLUT 4 seems to be absent, as a confirmation of other studies. Only in dog sperm cells capacitating conditions induce a change in GLUTs distribution, even if the physiological role of these changes should be deepened.
Gli spermatozoi di mammifero necessitano di zuccheri per espletare le loro funzioni, come il mantenimento dell’omeostasi membranale ed il movimento. Queste cellule hanno un metabolismo particolare, che non è ancora stato del tutto compreso, anche se è chiaro che ottengano energia da esosi tramite il processo glicolitico e la fosforilazione ossidativa. Gli spermatozoi sono a contatto con ambienti esterni molto diversi: dai fluidi testicolare ed epididimale, per passare al plasma seminale ed infine alle secrezioni dell’apparato genitale femminile. Inoltre, con la diffusione delle biotecnologie riproduttive, il liquido seminale è diluito e conservato in svariati media contenenti diversi substrati energetici. Per sfruttare queste fonti energetiche gli spermatozoi, come le altre cellule eucariotiche, hanno un sistema proteico di membrana ben definito, rappresentato principalmente dalla famiglia dei GLUT. Queste proteine hanno una struttura transmembranale ad alfa elica e funzionano come una pompa enzimatica che permette un trasporto passivo veloce e secondo gradiente di concentrazione delle molecole di zucchero attraverso lo strato lipidico. Alcuni GLUT sono stati studiati negli spermatozoi di uomo, ratto e toro e la presenza di alcuni altri è stata dimostrata in cane e maiale. Gli scopi di questo studio sono stati: • determinare la presenza dei GLUT 1, 2, 3, 4 e 5 in spermatozoi di maiale, cane , stallone e asino e descrivere la loro localizzazione; • studiare eventuali cambi di localizzazione dovuti alla capacitazione o alla reazione acrosomiale in spermatozoi di maiale, cane e stallone; • valutare cambiamenti nella disposizione dei GLUT negli spermatozoi di maiale in seguito a capacitazione stimolata con insulina ed IGF; • valutare possibili cambiamenti di localizzazione dei GLUT a seguito del processo di “sessaggio” mediante citofluorimetro sorter. La presenza e la localizzazione dei GLUT 1, 2, 3 e 5 sono state dimostrate negli spermatozoi di maiale, asino, cavallo e cane mediante le tecniche di western blotting ed immunofluorescenza indiretta; una rilocalizzazione delle proteine dopo capacitazione è stata osservata solo negli spermatozoi di cane e nessun cambiamento è stato registrato nelle altre specie. Per quanto riguarda il maiale, non si sono rilevate rilocalizzazioni dei GLUT a seguito della capacitazione con stimolazione con insulina ed IGF e nemmeno a seguito del processo di “sex sorting”. Concludendo, questo studio conferma la presenza dei GLUT 1, 2, 3 e 5 negli spermatozoi di maiale, cane stallone e asino, mentre il GLUT 4 sembra essere assente, a conferma di alcuni studi precedenti. Solo negli spermatozoi di cane le condizioni capacitanti inducono un cambiamento nella distribuzione dei GLUT, anche se il ruolo fisiologico di questi cambiamenti deve essere ancora approfondito.

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12

Tralli, Manuela <1979&gt. "Progetto Rivivere.Pet: il servizio on-line dell'Università di Bologna in supporto ai proprietari in lutto per la morte dell'animale." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2781/1/Tralli_Manuela_Tesi.pdf.

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13

Tralli, Manuela <1979&gt. "Progetto Rivivere.Pet: il servizio on-line dell'Università di Bologna in supporto ai proprietari in lutto per la morte dell'animale." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2781/.

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14

Francesconi, Mirko <1981&gt. "Edge perturbation in the transcription network of yeast." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2835/3/francesconi_mirko_tesi.pdf.

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15

Francesconi, Mirko <1981&gt. "Edge perturbation in the transcription network of yeast." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2835/.

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16

Raccagni, Ramona <1983&gt. "Aspetti fisiopatologici della prostata di cane." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3631/1/raccagni_ramona_tesi.pdf.

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Abstract:

Il lavoro eseguito comprende 3 differenti studi inerenti l’approfondimento di diversi aspetti della fisiopatologia della prostata, in particolare l’aspetto clinico, biochimico ed ormonale. Per quanto riguarda l’aspetto clinico è stato eseguito uno studio ecografico sull’accrescimento e sviluppo della ghiandola, a partire dai 4 giorni di vita all’anno di età del cane. L’aspetto biochimico dell’organo è stato indagato nel secondo studio, analizzando l’eventuali differenze nel profilo proteico e nel profilo dello zinco, valutato con spettrofotometria ad assorbimento atomico, su campioni di plasma seminale di cani sani o cani affetti da iperplasia prostatica benigna sintomatica od asintomatica. Lo studio è stato eseguito allo scopo di valutare se l’approccio biochimico ai problemi prostatici possa essere d’aiuto nella diagnosi della più comune patologia della prostata, l’iperplasia prostatica benigna. Il terzo studio inerente l’aspetto ormonale della prostata, completa lo studio precedente, analizzando le eventuali differenze ormonali (Testosterone, Estrogeni e Progesterone) riscontrabili fra due gruppi di cani: i cani sani e quelli affetti da iperplasia prostatica benigna.

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17

Raccagni, Ramona <1983&gt. "Aspetti fisiopatologici della prostata di cane." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3631/.

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Abstract:

Il lavoro eseguito comprende 3 differenti studi inerenti l’approfondimento di diversi aspetti della fisiopatologia della prostata, in particolare l’aspetto clinico, biochimico ed ormonale. Per quanto riguarda l’aspetto clinico è stato eseguito uno studio ecografico sull’accrescimento e sviluppo della ghiandola, a partire dai 4 giorni di vita all’anno di età del cane. L’aspetto biochimico dell’organo è stato indagato nel secondo studio, analizzando l’eventuali differenze nel profilo proteico e nel profilo dello zinco, valutato con spettrofotometria ad assorbimento atomico, su campioni di plasma seminale di cani sani o cani affetti da iperplasia prostatica benigna sintomatica od asintomatica. Lo studio è stato eseguito allo scopo di valutare se l’approccio biochimico ai problemi prostatici possa essere d’aiuto nella diagnosi della più comune patologia della prostata, l’iperplasia prostatica benigna. Il terzo studio inerente l’aspetto ormonale della prostata, completa lo studio precedente, analizzando le eventuali differenze ormonali (Testosterone, Estrogeni e Progesterone) riscontrabili fra due gruppi di cani: i cani sani e quelli affetti da iperplasia prostatica benigna.

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18

Li, Destri Nicosia Dora <1974&gt. "Equine and human mutual welfare: a whole subject? Critical aspects and possible strategies in equine-assisted activities and therapies." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3819/1/Li_Destri_Nicosia_Dora_tesi.pdf.

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General aim of the study is equine welfare, particularly concerning different husbandry methodic and inter-specific relational factors. Specific aim is the evaluation of possible mutual (to humans and to equines) benefits and the analysis of critical factors/strength points, of human-horse relationship within Therapeutic Riding context (TR). The peculiarities of human-horse relationship (compared to the bond with “Pet”) are analyzed, concerning their socio-anthropological, psychological, psycho-dynamic distinctive characteristics. 8 European representative therapeutic riding centers (TRC) were therefore selected (on the basis of their different animals’ husbandry criteria, and of the different rehabilitative methodologies adopted). TRC were investigated through 2 different questionnaires, specifically settled to access objective/subjective animal welfare parameters; the quality of human-horse relationship; technicians’ emotional experienced. 3 Centers were further selected, and behavioral (145 hours of behavioral recording) and physiological parameters (heart rate and heart rate variability) were evaluated, aimed to access equine welfare and horses’ adaptive responses/coping (towards general environment and towards TR job). Moreover a specific “handling-task” was ideated and experimented, aimed to measure the quality of TR technicians-horses relationship. We did therefore evaluate both the individual horses’ responses and the possible differences among Centers. Data collected highlight the lack of univocal standardized methodic, concerning the general animals’ management and the specific methodologies (aimed to improve animal welfare and to empower TR efficacy). Some positive and some critical aspects were detected concerning TR personnel-horse relationship. Another experimental approach did evaluate the efficacy (concerning the mutual benefits’ empowerment) of an “ethologically-fitted” TR intervention, aimed to educate children to and through the relationship with horses. Our data evidenced that the improvement of human horse relationship, through structured educational programs for TR personnel might have important consequences both to human and equine welfare.

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19

Li, Destri Nicosia Dora <1974&gt. "Equine and human mutual welfare: a whole subject? Critical aspects and possible strategies in equine-assisted activities and therapies." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3819/.

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Abstract:

General aim of the study is equine welfare, particularly concerning different husbandry methodic and inter-specific relational factors. Specific aim is the evaluation of possible mutual (to humans and to equines) benefits and the analysis of critical factors/strength points, of human-horse relationship within Therapeutic Riding context (TR). The peculiarities of human-horse relationship (compared to the bond with “Pet”) are analyzed, concerning their socio-anthropological, psychological, psycho-dynamic distinctive characteristics. 8 European representative therapeutic riding centers (TRC) were therefore selected (on the basis of their different animals’ husbandry criteria, and of the different rehabilitative methodologies adopted). TRC were investigated through 2 different questionnaires, specifically settled to access objective/subjective animal welfare parameters; the quality of human-horse relationship; technicians’ emotional experienced. 3 Centers were further selected, and behavioral (145 hours of behavioral recording) and physiological parameters (heart rate and heart rate variability) were evaluated, aimed to access equine welfare and horses’ adaptive responses/coping (towards general environment and towards TR job). Moreover a specific “handling-task” was ideated and experimented, aimed to measure the quality of TR technicians-horses relationship. We did therefore evaluate both the individual horses’ responses and the possible differences among Centers. Data collected highlight the lack of univocal standardized methodic, concerning the general animals’ management and the specific methodologies (aimed to improve animal welfare and to empower TR efficacy). Some positive and some critical aspects were detected concerning TR personnel-horse relationship. Another experimental approach did evaluate the efficacy (concerning the mutual benefits’ empowerment) of an “ethologically-fitted” TR intervention, aimed to educate children to and through the relationship with horses. Our data evidenced that the improvement of human horse relationship, through structured educational programs for TR personnel might have important consequences both to human and equine welfare.

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20

De, Cecco Marco <1984&gt. "Genetic Manipulation, Gene Silencing and Biomarker Development in Multiple Experimental Models." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2012. http://amsdottorato.unibo.it/4254/1/DeCecco_Marco_tesi.pdf.

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Abstract:

The thesis is set in three different parts, according to the relative experimental models. First, the domestic pig (Sus scrofa) is part of the study on reproductive biotechnologies: the transgenesis technique of Sperm Mediated Gene Transfer is widely studied starting from the quality of the semen, through the study of multiple uptakes of exogenous DNA and lastly used in the production of multi-transgenic blastocysts. Finally we managed to couple the transgenesis pipeline with sperm sorting and therefore produced transgenic embryos of predetermined sex. In the second part of the thesis the attention is on the fruit fly (Drosophila melanogaster) and on its derived cell line: the S2 cells. The in vitro and in vivo models are used to develop and validate an efficient way to knock down the myc gene. First an efficient in vitro protocol is described, than we demonstrate how the decrease in myc transcript remarkably affects the ribosome biogenesis through the study of Polysome gradients, rRNA content and qPCR. In vivo we identified two optimal drivers for the conditional silencing of myc, once the flies are fed with RU486: the first one is throughout the whole body (Tubulin), while the second is a head fat body driver (S32). With these results we present a very efficient model to study the role of myc in multiple aspects of translation. In the third and last part, the focus is on human derived lung fibroblasts (hLF-1), mouse tail fibroblasts and mouse tissues. We developed an efficient assay to quantify the total protein content of the nucleus on a single cell level via fluorescence. We coupled the protocol with classical immunofluorescence so to have at the same time general and particular information, demonstrating that during senescence nuclear proteins increase by 1.8 fold either in human cells, mouse cells and mouse tissues.

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21

De, Cecco Marco <1984&gt. "Genetic Manipulation, Gene Silencing and Biomarker Development in Multiple Experimental Models." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2012. http://amsdottorato.unibo.it/4254/.

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Abstract:

The thesis is set in three different parts, according to the relative experimental models. First, the domestic pig (Sus scrofa) is part of the study on reproductive biotechnologies: the transgenesis technique of Sperm Mediated Gene Transfer is widely studied starting from the quality of the semen, through the study of multiple uptakes of exogenous DNA and lastly used in the production of multi-transgenic blastocysts. Finally we managed to couple the transgenesis pipeline with sperm sorting and therefore produced transgenic embryos of predetermined sex. In the second part of the thesis the attention is on the fruit fly (Drosophila melanogaster) and on its derived cell line: the S2 cells. The in vitro and in vivo models are used to develop and validate an efficient way to knock down the myc gene. First an efficient in vitro protocol is described, than we demonstrate how the decrease in myc transcript remarkably affects the ribosome biogenesis through the study of Polysome gradients, rRNA content and qPCR. In vivo we identified two optimal drivers for the conditional silencing of myc, once the flies are fed with RU486: the first one is throughout the whole body (Tubulin), while the second is a head fat body driver (S32). With these results we present a very efficient model to study the role of myc in multiple aspects of translation. In the third and last part, the focus is on human derived lung fibroblasts (hLF-1), mouse tail fibroblasts and mouse tissues. We developed an efficient assay to quantify the total protein content of the nucleus on a single cell level via fluorescence. We coupled the protocol with classical immunofluorescence so to have at the same time general and particular information, demonstrating that during senescence nuclear proteins increase by 1.8 fold either in human cells, mouse cells and mouse tissues.

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22

Vallorani, Claudia <1983&gt. "Conservazione del seme sortato di suino." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2012. http://amsdottorato.unibo.it/4550/1/Vallorani_Claudia_tesi.pdf.

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Abstract:

Gli spermatozoi di suino sottoposti alla procedura di sessaggio mediante citofluorimetria presentano una serie di modificazioni morfo-funzionali che compromettono nel tempo la loro sopravvivenza e la capacità fecondante. Questi spermatozoi, inoltre, a causa della sensibilità ai danni indotti dalla crioconservazione, vengono solitamente conservati allo stato liquido a 15-17°C, con conseguente ulteriore peggioramento nel tempo della qualità delle cellule spermatiche sessate. Lo scopo della ricerca è stato quello di valutare le modificazioni di alcune caratteristiche morfo-funzionali degli spermatozoi in seguito a sex-sorting e conseguente conservazione. Successivamente si è cercato di migliorare i parametri qualitativi del seme sessato mediante l’aggiunta di sostanze antiossidanti e la messa a punto di una nuova metodica di conservazione. I risultati ottenuti hanno evidenziato che la procedura di sessaggio e la conseguente conservazione per 24-26 ore a 15°C hanno indotto un peggioramento significativo delle caratteristiche morfo-funzionali (vitalità, integrità acrosomiale, quantità e distribuzione dell’Hsp70, capacità fecondante). Mentre l’azione degli antiossidanti non si è rivelata efficace nel miglioramento della qualità degli spermatozoi durante le fasi di colorazione e passaggio attraverso il citofluorimetro, l’azione congiunta del plasma seminale e degli antiossidanti superossido-dismutasi ed epigallocatechina-3-gallato ha indotto un miglioramento significativo della vitalità degli spermatozoi. Per la conservazione del seme di suino è stata testata la tecnica di incapsulazione in membrane di alginato di bario che permette, durante l’inseminazione artificiale, un rilascio graduale degli spermatozoi e l’utilizzo di un quantitativo inferiore di materiale seminale. L’applicazione di tale tecnica per la conservazione degli spermatozoi di suino sessati non sembra provocare un calo significativo della vitalità, dell’integrità acrosomiale e dell’efficienza totale di fecondazione rispetto al seme sortato e conservato diluito suggerendo futuri studi in vivo. Una migliore conoscenza dei danni indotti da queste tecnologie e la loro minimizzazione potrà stimolare in futuro l’utilizzo su vasta scala del seme sessato nel suino.
Boar spermatozoa submitted to the sorting procedure show several morpho-functional modifications effective in compromising their survival and fertilization ability. Moreover ,boar spermatozoa, because of their susceptibility to damages induced by cryopreservation, are usually stored at 15-17°C after the sorting procedure; however, also the conservation at liquid state implies the worsening of semen quality. The aims of this research were: 1) to evaluate morpho-functional characteristics of sperm cells submitted to sex-sorting and consequent storage; 2) to try to improve the quality of sorted semen by the addition of antioxidants; 2) to set up a new storage method. Our results evidence a decreased quality of boar sorted-stored spermatozoa in term of: viability, acrosome integrity, amount and localization of Hsp70, fertilizing ability. During the staining step and the passage through the cytofluorimeter, antioxidants were not effective in improving sperm cells morpho-functional characteristics, while the addition of superoxide dismutase or epigallocatechin-3-gallate associated with seminal plasma induced an increase of viability of sorted boar spermatozoa stored 24 h at 15°C. Some researchers have utilized encapsulation in barium alginate membrane to store boar sperm cells. This technique allows a constant release of spermatozoa in sow reproductive system, avoiding the double/triple intervention of insemination and reducing the number of spermatozoa/insemination. The application of this technique in order to store boar sperm cells after sorting did not induce any impairment of sperm morpho-functional characteristics (viability, acrosome integrity, total efficiency of insemination) compared to sorted spermatozoa stored at liquid state, thus demonstrating the possibility to use this method to improve the reproductive performance of boar sorted semen.

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23

Vallorani, Claudia <1983&gt. "Conservazione del seme sortato di suino." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2012. http://amsdottorato.unibo.it/4550/.

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Abstract:

Gli spermatozoi di suino sottoposti alla procedura di sessaggio mediante citofluorimetria presentano una serie di modificazioni morfo-funzionali che compromettono nel tempo la loro sopravvivenza e la capacità fecondante. Questi spermatozoi, inoltre, a causa della sensibilità ai danni indotti dalla crioconservazione, vengono solitamente conservati allo stato liquido a 15-17°C, con conseguente ulteriore peggioramento nel tempo della qualità delle cellule spermatiche sessate. Lo scopo della ricerca è stato quello di valutare le modificazioni di alcune caratteristiche morfo-funzionali degli spermatozoi in seguito a sex-sorting e conseguente conservazione. Successivamente si è cercato di migliorare i parametri qualitativi del seme sessato mediante l’aggiunta di sostanze antiossidanti e la messa a punto di una nuova metodica di conservazione. I risultati ottenuti hanno evidenziato che la procedura di sessaggio e la conseguente conservazione per 24-26 ore a 15°C hanno indotto un peggioramento significativo delle caratteristiche morfo-funzionali (vitalità, integrità acrosomiale, quantità e distribuzione dell’Hsp70, capacità fecondante). Mentre l’azione degli antiossidanti non si è rivelata efficace nel miglioramento della qualità degli spermatozoi durante le fasi di colorazione e passaggio attraverso il citofluorimetro, l’azione congiunta del plasma seminale e degli antiossidanti superossido-dismutasi ed epigallocatechina-3-gallato ha indotto un miglioramento significativo della vitalità degli spermatozoi. Per la conservazione del seme di suino è stata testata la tecnica di incapsulazione in membrane di alginato di bario che permette, durante l’inseminazione artificiale, un rilascio graduale degli spermatozoi e l’utilizzo di un quantitativo inferiore di materiale seminale. L’applicazione di tale tecnica per la conservazione degli spermatozoi di suino sessati non sembra provocare un calo significativo della vitalità, dell’integrità acrosomiale e dell’efficienza totale di fecondazione rispetto al seme sortato e conservato diluito suggerendo futuri studi in vivo. Una migliore conoscenza dei danni indotti da queste tecnologie e la loro minimizzazione potrà stimolare in futuro l’utilizzo su vasta scala del seme sessato nel suino.
Boar spermatozoa submitted to the sorting procedure show several morpho-functional modifications effective in compromising their survival and fertilization ability. Moreover ,boar spermatozoa, because of their susceptibility to damages induced by cryopreservation, are usually stored at 15-17°C after the sorting procedure; however, also the conservation at liquid state implies the worsening of semen quality. The aims of this research were: 1) to evaluate morpho-functional characteristics of sperm cells submitted to sex-sorting and consequent storage; 2) to try to improve the quality of sorted semen by the addition of antioxidants; 2) to set up a new storage method. Our results evidence a decreased quality of boar sorted-stored spermatozoa in term of: viability, acrosome integrity, amount and localization of Hsp70, fertilizing ability. During the staining step and the passage through the cytofluorimeter, antioxidants were not effective in improving sperm cells morpho-functional characteristics, while the addition of superoxide dismutase or epigallocatechin-3-gallate associated with seminal plasma induced an increase of viability of sorted boar spermatozoa stored 24 h at 15°C. Some researchers have utilized encapsulation in barium alginate membrane to store boar sperm cells. This technique allows a constant release of spermatozoa in sow reproductive system, avoiding the double/triple intervention of insemination and reducing the number of spermatozoa/insemination. The application of this technique in order to store boar sperm cells after sorting did not induce any impairment of sperm morpho-functional characteristics (viability, acrosome integrity, total efficiency of insemination) compared to sorted spermatozoa stored at liquid state, thus demonstrating the possibility to use this method to improve the reproductive performance of boar sorted semen.

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24

Beghelli, Valentina <1982&gt. "Studio di alcuni tratti del temperamento nel cane." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6239/1/beghelli_valentina_tesi.pdf.

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Abstract:

Il temperamento può essere definito come l’attitudine che un cane esprime verso le persone e verso altri animali, la combinazione di tratti fisici e mentali, acquisiti e non, che determinano il comportamento del cane. Tale parametro delinea perciò il carattere di un individuo, inclinazioni e tendenze, eccitabilità, tristezza, rabbia e il modo caratteristico di comportarsi di un individuo, con particolare riferimento alle interazioni sociali. La presente tesi di Dottorato rappresenta uno studio su alcuni tratti del temperamento nel cane domestico elaborato in 3 progetti sperimentali. Nei primi due progetti sono state analizzate le differenze attitudinali tra alcune razze canine attraverso l’applicazione di un test di temperamento in cuccioli di 60 giorni e in cani adulti, per valutare e confrontarne il temperamento, la socialità ed identificare profili tipici di razza. Nel terzo progetto un campione di cani morsicatori di canile e di proprietà è stato confrontato con due rispettivi gruppi di controllo. Analizzando i risultati è stato possibile mettere in evidenza caratteristiche di razza omogenee nell’interazione con stimoli inanimati, nelle interazioni sociali e in relazione alla possessività e sono stati delineati profili di razza sia nei cuccioli sia negli adulti. Si sono tuttavia, osservate variabilità individuali, intra-razza e intra-cucciolata, a testimonianza dell’influenza complessa e multifattoriale delle componenti genetica e ambientale sul comportamento dei cani. Il confronto tra cani morsicatori di canile e di proprietà ha messo in luce interessanti differenze tra i soggetti in termini di reattività, socievolezza, propensione all’interazione con il proprietario o con un estraneo, comportamenti di evitamento e velocità di reazione agli stimoli presentati. Il test applicato è risultato un valido strumento per valutare il temperamento di cani dichiarati aggressivi che sono stati sottoposti a situazioni nuove e a stimoli sconosciuti per poter ottenere una migliore visione d’insieme del temperamento del soggetto.
Temperament has been defined as the attitude expressed by the dog toward people or other animals, the combination of physical and mental traits, acquired or not, which determine the behavior of the dog and therefore delineates the character of an individual, his inclinations, trends and the characteristic way of behaving, with particular reference to social interactions. This work represents a study of temperamental traits in the Domestic Dog, developed in 3 experimental projects. In the first two projects the attitudinal differences between some dog breeds are analyzed, through the application of a temperament test in puppies 60 days old and adult, in order to evaluate and compare their temperament and sociability and to identify specific breed profiles. In the third project, a sample of biters dogs coming from different kennel of and pet biters dogs were tested and compared with two respective control groups. Analyzing the results, it was possible to highlight consistent breed characteristics in the interaction with inanimate stimuli, in social interactions and in relation to possessivness and a first breed profile has been outlined for both puppies and adults. However, individual variabilities were observed, relative to breed and litter, reflecting the complex and multifactorial influence of genetic and environmental components on the behavior of the dog. Furthermore, the study allowed us to evaluate dogs biters and compare them with the control groups, on the base of temperament traits. The comparison between biters coming from kennels and pet ones underlined differences in terms of reactivity, sociability, attitude to interact with the owner or with a stranger, in avoiding trends and velocity of reaction to the stimuli presented . The temperament test can be considered a valid tool to assess the temperament of aggressive dogs, in order to get a better overview of the temperament of the subjects.

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25

Beghelli, Valentina <1982&gt. "Studio di alcuni tratti del temperamento nel cane." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6239/.

Full text

Abstract:

Il temperamento può essere definito come l’attitudine che un cane esprime verso le persone e verso altri animali, la combinazione di tratti fisici e mentali, acquisiti e non, che determinano il comportamento del cane. Tale parametro delinea perciò il carattere di un individuo, inclinazioni e tendenze, eccitabilità, tristezza, rabbia e il modo caratteristico di comportarsi di un individuo, con particolare riferimento alle interazioni sociali. La presente tesi di Dottorato rappresenta uno studio su alcuni tratti del temperamento nel cane domestico elaborato in 3 progetti sperimentali. Nei primi due progetti sono state analizzate le differenze attitudinali tra alcune razze canine attraverso l’applicazione di un test di temperamento in cuccioli di 60 giorni e in cani adulti, per valutare e confrontarne il temperamento, la socialità ed identificare profili tipici di razza. Nel terzo progetto un campione di cani morsicatori di canile e di proprietà è stato confrontato con due rispettivi gruppi di controllo. Analizzando i risultati è stato possibile mettere in evidenza caratteristiche di razza omogenee nell’interazione con stimoli inanimati, nelle interazioni sociali e in relazione alla possessività e sono stati delineati profili di razza sia nei cuccioli sia negli adulti. Si sono tuttavia, osservate variabilità individuali, intra-razza e intra-cucciolata, a testimonianza dell’influenza complessa e multifattoriale delle componenti genetica e ambientale sul comportamento dei cani. Il confronto tra cani morsicatori di canile e di proprietà ha messo in luce interessanti differenze tra i soggetti in termini di reattività, socievolezza, propensione all’interazione con il proprietario o con un estraneo, comportamenti di evitamento e velocità di reazione agli stimoli presentati. Il test applicato è risultato un valido strumento per valutare il temperamento di cani dichiarati aggressivi che sono stati sottoposti a situazioni nuove e a stimoli sconosciuti per poter ottenere una migliore visione d’insieme del temperamento del soggetto.
Temperament has been defined as the attitude expressed by the dog toward people or other animals, the combination of physical and mental traits, acquired or not, which determine the behavior of the dog and therefore delineates the character of an individual, his inclinations, trends and the characteristic way of behaving, with particular reference to social interactions. This work represents a study of temperamental traits in the Domestic Dog, developed in 3 experimental projects. In the first two projects the attitudinal differences between some dog breeds are analyzed, through the application of a temperament test in puppies 60 days old and adult, in order to evaluate and compare their temperament and sociability and to identify specific breed profiles. In the third project, a sample of biters dogs coming from different kennel of and pet biters dogs were tested and compared with two respective control groups. Analyzing the results, it was possible to highlight consistent breed characteristics in the interaction with inanimate stimuli, in social interactions and in relation to possessivness and a first breed profile has been outlined for both puppies and adults. However, individual variabilities were observed, relative to breed and litter, reflecting the complex and multifactorial influence of genetic and environmental components on the behavior of the dog. Furthermore, the study allowed us to evaluate dogs biters and compare them with the control groups, on the base of temperament traits. The comparison between biters coming from kennels and pet ones underlined differences in terms of reactivity, sociability, attitude to interact with the owner or with a stranger, in avoiding trends and velocity of reaction to the stimuli presented . The temperament test can be considered a valid tool to assess the temperament of aggressive dogs, in order to get a better overview of the temperament of the subjects.

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26

Peric, Tanja <1982&gt. "Hair: a tool to evaluate the HPA axis activity." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6253/1/WHOLE_THESIS_PERIC.pdf.

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Abstract:

Hair cortisol is a novel marker to measure long-term secretion cortisol free from many methodological caveats associated with other matrices such as plasma, saliva, urine, milk and faeces. For decades hair analysis has been successfully used in forensic science and toxicology to evaluate the exposure to exogenous substances and assess endogenous steroid hormones. Evaluation of cortisol in hair matrix began about a decade ago and have over the past five years had a remarkable development by advancing knowledge and affirming this method as a new and efficient way to study the hypothalamic-pituitary-adrenal (HPA) axis activity over a long time period. In farm animals, certain environmental or management conditions can potentially activate the HPA axis. Given the importance of cortisol in monitoring the HPA axis activity, a first approach has involved the study on the distribution of hair cortisol concentrations (HCC) in healthy dairy cows showing a physiological range of variation of this hormone. Moreover, HCC have been significantly influenced also by changes in environmental conditions and a significant positive correlation was detected between HCC and cows clinically or physiologically compromised suggesting that these cows were subjected to repeated HPA axis activation. Additionally, Crossbreed F1 heifers showed significantly lower HCC compared to pure animals and a breed influence has been seen also on the HPA axis activity stimulated by an environmental change showing thus a higher level of resilience and a better adaptability to the environment of certain genotypes. Hair proved to be an excellent matrix also in the study of the activation of the HPA axis during the perinatal period. The use of hair analysis in research holds great promise to significantly enhance current understanding on the role of HPA axis over a long period of time.

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27

Peric, Tanja <1982&gt. "Hair: a tool to evaluate the HPA axis activity." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6253/.

Full text

Abstract:

Hair cortisol is a novel marker to measure long-term secretion cortisol free from many methodological caveats associated with other matrices such as plasma, saliva, urine, milk and faeces. For decades hair analysis has been successfully used in forensic science and toxicology to evaluate the exposure to exogenous substances and assess endogenous steroid hormones. Evaluation of cortisol in hair matrix began about a decade ago and have over the past five years had a remarkable development by advancing knowledge and affirming this method as a new and efficient way to study the hypothalamic-pituitary-adrenal (HPA) axis activity over a long time period. In farm animals, certain environmental or management conditions can potentially activate the HPA axis. Given the importance of cortisol in monitoring the HPA axis activity, a first approach has involved the study on the distribution of hair cortisol concentrations (HCC) in healthy dairy cows showing a physiological range of variation of this hormone. Moreover, HCC have been significantly influenced also by changes in environmental conditions and a significant positive correlation was detected between HCC and cows clinically or physiologically compromised suggesting that these cows were subjected to repeated HPA axis activation. Additionally, Crossbreed F1 heifers showed significantly lower HCC compared to pure animals and a breed influence has been seen also on the HPA axis activity stimulated by an environmental change showing thus a higher level of resilience and a better adaptability to the environment of certain genotypes. Hair proved to be an excellent matrix also in the study of the activation of the HPA axis during the perinatal period. The use of hair analysis in research holds great promise to significantly enhance current understanding on the role of HPA axis over a long period of time.

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28

Montillo, Marta <1986&gt. "Hormonal parameters in foal hair: from birth to post weaning." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6254/1/Hormonal_parameters_in_foal_hair_from_birth_to_post_weaning.pdf.

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Abstract:

The aim of this study was to investigate cortisol and progesterone (P4) trends in hair from birth up to postweaning in Italian trotter foals. Hair sampling is non-invasive and hair concentrations provide retrospective information of integrated hormone secretion over periods of several months. Samples were collected at birth and at a distance of 30 days, collecting only regrowth hair, up to post weaning. From birth to 3 months, foals cortisol falls from 47.64±5.6 to 4.9±0.68 pg/mg (mean±standard error), due to the interruption of foetal-placental connection and progressive adaptation to extrauterine life. From the third month of life to post weaning concentrations don’t vary significantly, underlining a non-chronic activation of the HPA axis. Hair P4 significantly decreases in the first two samples (from 469.68±72,54 to 184.65±35.42 pg/mg). At 2 (111.78±37.13 pg/mg) and 3 months (35.96±6.33 pg/mg) hair concentrations don’t show significant differences. These concentrations are not due to interactions of the utero-placental tissues with foals, animals are still prepuberal and P4 isn’t produced by adrenals as a result of high stress. We could therefore hypothesize that the source of foal hair P4 could be milk, suckled from mares. The high individual variability in hair at 2 and 3 months is due to a gradual and subjective change in foal diet, from milk to solid food, and to the fact that mares do not allow to suckle. From fourth month to post weaning P4 concentration in hair remains around 37.56±6.45 pg/mg. In conclusion, hair collected at birth, giving information about last period of gestation, could be used along with traditional matrices, to evaluate foals maturity. Hair cortisol could give indications about foals capacity to adapt to extra-uterine life. Finally milk, configuring as a bringer of nutrients and energy and assuming the characteristic of a nutraceutical, could give fundamental information about parental care.

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29

Montillo, Marta <1986&gt. "Hormonal parameters in foal hair: from birth to post weaning." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6254/.

Full text

Abstract:

The aim of this study was to investigate cortisol and progesterone (P4) trends in hair from birth up to postweaning in Italian trotter foals. Hair sampling is non-invasive and hair concentrations provide retrospective information of integrated hormone secretion over periods of several months. Samples were collected at birth and at a distance of 30 days, collecting only regrowth hair, up to post weaning. From birth to 3 months, foals cortisol falls from 47.64±5.6 to 4.9±0.68 pg/mg (mean±standard error), due to the interruption of foetal-placental connection and progressive adaptation to extrauterine life. From the third month of life to post weaning concentrations don’t vary significantly, underlining a non-chronic activation of the HPA axis. Hair P4 significantly decreases in the first two samples (from 469.68±72,54 to 184.65±35.42 pg/mg). At 2 (111.78±37.13 pg/mg) and 3 months (35.96±6.33 pg/mg) hair concentrations don’t show significant differences. These concentrations are not due to interactions of the utero-placental tissues with foals, animals are still prepuberal and P4 isn’t produced by adrenals as a result of high stress. We could therefore hypothesize that the source of foal hair P4 could be milk, suckled from mares. The high individual variability in hair at 2 and 3 months is due to a gradual and subjective change in foal diet, from milk to solid food, and to the fact that mares do not allow to suckle. From fourth month to post weaning P4 concentration in hair remains around 37.56±6.45 pg/mg. In conclusion, hair collected at birth, giving information about last period of gestation, could be used along with traditional matrices, to evaluate foals maturity. Hair cortisol could give indications about foals capacity to adapt to extra-uterine life. Finally milk, configuring as a bringer of nutrients and energy and assuming the characteristic of a nutraceutical, could give fundamental information about parental care.

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30

Scorrano, Fabrizio <1985&gt. "Validation of a new method to assess the long-term Hypothalamic-Pituitary-Adrenal Activity using hair glucocorticoids as biomarkers. - Studies on pigs and laboratory Sprague-Dawley Rats." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6276/1/Fabrizio_Scorrano_TESI.pdf.

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Abstract:

The evaluation of chronic activity of the hypothalamic-pituitary-adrenal (HPA) axis is critical for determining the impact of chronic stressful situations. The potential use of hair glucocorticoids as a non-invasive, retrospective, biomarker of long term HPA activity is of great interest, and it is gaining acceptance in humans and animals. However, there are still no studies in literature examining hair cortisol concentration in pigs and corticosterone concentration in laboratory rodents. Therefore, we developed and validated, for the first time, a method for measuring hair glucocorticoids concentration in commercial sows and in Sprague-Dawley rats. Our preliminary data demonstrated: 1) a validated and specific washing protocol and extraction assay method with a good sensitivity in both species; 2) the effect of the reproductive phase, housing conditions and seasonality on hair cortisol concentration in sows; 3) similar hair corticosterone concentration in male and female rats; 4) elevated hair corticosterone concentration in response to chronic stress manipulations and chronic ACTH administration, demonstrating that hair provides a good direct index of HPA activity over long periods than other indirect parameters, such adrenal or thymus weight. From these results we believe that this new non-invasive tool needs to be applied to better characterize the overall impact in livestock animals and in laboratory rodents of chronic stressful situations that negatively affect animals welfare. Nevertheless, further studies are needed to improve this methodology and maybe to develop animal models for chronic stress of high interest and translational value in human medicine.

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31

Scorrano, Fabrizio <1985&gt. "Validation of a new method to assess the long-term Hypothalamic-Pituitary-Adrenal Activity using hair glucocorticoids as biomarkers. - Studies on pigs and laboratory Sprague-Dawley Rats." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6276/.

Full text

Abstract:

The evaluation of chronic activity of the hypothalamic-pituitary-adrenal (HPA) axis is critical for determining the impact of chronic stressful situations. The potential use of hair glucocorticoids as a non-invasive, retrospective, biomarker of long term HPA activity is of great interest, and it is gaining acceptance in humans and animals. However, there are still no studies in literature examining hair cortisol concentration in pigs and corticosterone concentration in laboratory rodents. Therefore, we developed and validated, for the first time, a method for measuring hair glucocorticoids concentration in commercial sows and in Sprague-Dawley rats. Our preliminary data demonstrated: 1) a validated and specific washing protocol and extraction assay method with a good sensitivity in both species; 2) the effect of the reproductive phase, housing conditions and seasonality on hair cortisol concentration in sows; 3) similar hair corticosterone concentration in male and female rats; 4) elevated hair corticosterone concentration in response to chronic stress manipulations and chronic ACTH administration, demonstrating that hair provides a good direct index of HPA activity over long periods than other indirect parameters, such adrenal or thymus weight. From these results we believe that this new non-invasive tool needs to be applied to better characterize the overall impact in livestock animals and in laboratory rodents of chronic stressful situations that negatively affect animals welfare. Nevertheless, further studies are needed to improve this methodology and maybe to develop animal models for chronic stress of high interest and translational value in human medicine.

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32

Mattioli, Michela <1975&gt. "Studio longitudinale sulle capacità cognitive del cane: discriminazione di quantità ed apprendimento per imitazione in relazione all'attaccamento." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6614/1/MATTIOLI_MICHELA_TESI.pdf.

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Abstract:

Il presente studio ha indagato e valutato alcune abilità cognitive del cane: la capacità di discriminare quantità e le capacità di apprendimento mediante imitazione; quest’ultima è poi stata messa in relazione con l’attaccamento nei confronti del proprietario. Per l’esecuzione della prima indagine sono stati messi appunto due test: il primo si è basato esclusivamente sulla presentazione di uno stimolo visivo: diversi quantitativi di cibo, differenti tra loro del 50%, sono stati presentati al cane; la scelta effettuata dai soggetti testati è stata premiata con differenti tipi di rinforzo differenziale o non differenziale. Il secondo test è stato diviso in due parti: sono stati presentati al cane diversi quantitativi di cibo sempre differenti tra loro del 50% ma nella prima parte del test l’input sensoriale per il cane è stato esclusivamente uditivo mentre nella seconda parte è stato sia uditivo che visivo. Ove è stato possibile è stato applicato ai cani un cardiofrequenzimetro al fine di eseguire una valutazione delle variazioni della frequenza cardiaca nel corso del test. Lo scopo è stato quello di valutare se i soggetti testati erano in grado di discriminare la quantità maggiore. La seconda indagine ha analizzato le capacità di apprendimento di 36 soggetti che sono stati suddivisi in cani da lavoro e pet. I soggetti protagonisti dello studio hanno eseguito il Mirror Test per la valutazione dell’apprendimento per imitazione. I soggetti presi in considerazione, sono stati sottoposti a scansione termografica all’inizio ed al termine del test ed è stata rilevata la loro frequenza respiratoria nella fase iniziale e finale del test. In 11 soggetti che hanno eseguito il precedente test è stato possibile eseguire anche il Strange Situation Test per la valutazione dell’attaccamento al proprietario; i test in questione sono stati videoregistrati ed analizzati per mezzo di un software preposto (OBSERVER XT 10).
The present study investigated and evaluated some cognitive abilities of the dog: the ability to discriminate quantity and to learn through imitation; the latter kind of learning was related to the dog’s attachment to the owner. In the first analysis two tests were applied: the first is based on the presentation of a visual stimulus: different amounts of food, different from each other by 50% , were presented to the dog; the choice of the dogs was rewarded with different types of differential or non-differential reinforcement. The second test was divided into two parts: different quantities of food (different from each other by 50 %) were submitted to the dogs; in the first part of the test, the sensory input was exclusively auditory, while in the second part both auditory and visual. Where feasible, a heart rate monitor was applied to the dogs, in order to perform an assessment of the heart rate changes during the test. The aim of the test was to evaluate if the subjects were able to discriminate the larger amount of food. The second study analyzed the learning ability of 36 subjects, divided into working dogs and pets. The subjects performed the Mirror Test for the assessment of learning by imitation. In addition the subjects were subjected to thermal scanning at the beginning and at the end of the test and their respiratory rate was detected before and after the test. Finally, in 11 of these subjects the Strange Situation Test for the assessment of attachment to the owner, was performed. All tests were videotaped and then analyzed using a dedicated software (OBSERVER XT 10).

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33

Mattioli, Michela <1975&gt. "Studio longitudinale sulle capacità cognitive del cane: discriminazione di quantità ed apprendimento per imitazione in relazione all'attaccamento." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2014. http://amsdottorato.unibo.it/6614/.

Full text

Abstract:

Il presente studio ha indagato e valutato alcune abilità cognitive del cane: la capacità di discriminare quantità e le capacità di apprendimento mediante imitazione; quest’ultima è poi stata messa in relazione con l’attaccamento nei confronti del proprietario. Per l’esecuzione della prima indagine sono stati messi appunto due test: il primo si è basato esclusivamente sulla presentazione di uno stimolo visivo: diversi quantitativi di cibo, differenti tra loro del 50%, sono stati presentati al cane; la scelta effettuata dai soggetti testati è stata premiata con differenti tipi di rinforzo differenziale o non differenziale. Il secondo test è stato diviso in due parti: sono stati presentati al cane diversi quantitativi di cibo sempre differenti tra loro del 50% ma nella prima parte del test l’input sensoriale per il cane è stato esclusivamente uditivo mentre nella seconda parte è stato sia uditivo che visivo. Ove è stato possibile è stato applicato ai cani un cardiofrequenzimetro al fine di eseguire una valutazione delle variazioni della frequenza cardiaca nel corso del test. Lo scopo è stato quello di valutare se i soggetti testati erano in grado di discriminare la quantità maggiore. La seconda indagine ha analizzato le capacità di apprendimento di 36 soggetti che sono stati suddivisi in cani da lavoro e pet. I soggetti protagonisti dello studio hanno eseguito il Mirror Test per la valutazione dell’apprendimento per imitazione. I soggetti presi in considerazione, sono stati sottoposti a scansione termografica all’inizio ed al termine del test ed è stata rilevata la loro frequenza respiratoria nella fase iniziale e finale del test. In 11 soggetti che hanno eseguito il precedente test è stato possibile eseguire anche il Strange Situation Test per la valutazione dell’attaccamento al proprietario; i test in questione sono stati videoregistrati ed analizzati per mezzo di un software preposto (OBSERVER XT 10).
The present study investigated and evaluated some cognitive abilities of the dog: the ability to discriminate quantity and to learn through imitation; the latter kind of learning was related to the dog’s attachment to the owner. In the first analysis two tests were applied: the first is based on the presentation of a visual stimulus: different amounts of food, different from each other by 50% , were presented to the dog; the choice of the dogs was rewarded with different types of differential or non-differential reinforcement. The second test was divided into two parts: different quantities of food (different from each other by 50 %) were submitted to the dogs; in the first part of the test, the sensory input was exclusively auditory, while in the second part both auditory and visual. Where feasible, a heart rate monitor was applied to the dogs, in order to perform an assessment of the heart rate changes during the test. The aim of the test was to evaluate if the subjects were able to discriminate the larger amount of food. The second study analyzed the learning ability of 36 subjects, divided into working dogs and pets. The subjects performed the Mirror Test for the assessment of learning by imitation. In addition the subjects were subjected to thermal scanning at the beginning and at the end of the test and their respiratory rate was detected before and after the test. Finally, in 11 of these subjects the Strange Situation Test for the assessment of attachment to the owner, was performed. All tests were videotaped and then analyzed using a dedicated software (OBSERVER XT 10).

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34

Zaniboni, Andrea <1986&gt. "The porcine animal model goes through the 3Rs: development of in vitro and ex vivo system to study vascular biology." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amsdottorato.unibo.it/6834/1/Zaniboni_Andrea_tesi.pdf.

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Since the publication of the book of Russell and Burch in 1959, scientific research has never stopped improving itself with regard to the important issue of animal experimentation. The European Directive 2010/63/EU “On the protection of animals used for scientific purposes” focuses mainly on the animal welfare, fixing the Russell and Burch’s 3Rs principles as the foundations of the document. In particular, the legislator clearly states the responsibility of the scientific community to improve the number of alternative methods to animal experimentation. The swine is considered a species of relevant interest for translational research and medicine due to its biological similarities with humans. The surgical community has, in fact, recognized the swine as an excellent model replicating the human cardiovascular system. There have been several wild-type and transgenic porcine models which were produced for biomedicine and translational research. Among these, the cardiovascular ones are the most represented. The continuous involvement of the porcine animal model in the biomedical research, as the continuous advances achieved using swine in translational medicine, support the need for alternative methods to animal experimentation involving pigs. The main purpose of the present work was to develop and characterize novel porcine alternative methods for cardiovascular translational biology/medicine. The work was mainly based on two different models: the first consisted in an ex vivo culture of porcine aortic cylinders and the second consisted in an in vitro culture of porcine aortic derived progenitor cells. Both the models were properly characterized and results indicated that they could be useful to the study of vascular biology. Nevertheless, both the models aim to reduce the use of experimental animals and to refine animal based-trials. In conclusion, the present research aims to be a small, but significant, contribution to the important and necessary field of study of alternative methods to animal experimentation.

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Zaniboni, Andrea <1986&gt. "The porcine animal model goes through the 3Rs: development of in vitro and ex vivo system to study vascular biology." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amsdottorato.unibo.it/6834/.

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Since the publication of the book of Russell and Burch in 1959, scientific research has never stopped improving itself with regard to the important issue of animal experimentation. The European Directive 2010/63/EU “On the protection of animals used for scientific purposes” focuses mainly on the animal welfare, fixing the Russell and Burch’s 3Rs principles as the foundations of the document. In particular, the legislator clearly states the responsibility of the scientific community to improve the number of alternative methods to animal experimentation. The swine is considered a species of relevant interest for translational research and medicine due to its biological similarities with humans. The surgical community has, in fact, recognized the swine as an excellent model replicating the human cardiovascular system. There have been several wild-type and transgenic porcine models which were produced for biomedicine and translational research. Among these, the cardiovascular ones are the most represented. The continuous involvement of the porcine animal model in the biomedical research, as the continuous advances achieved using swine in translational medicine, support the need for alternative methods to animal experimentation involving pigs. The main purpose of the present work was to develop and characterize novel porcine alternative methods for cardiovascular translational biology/medicine. The work was mainly based on two different models: the first consisted in an ex vivo culture of porcine aortic cylinders and the second consisted in an in vitro culture of porcine aortic derived progenitor cells. Both the models were properly characterized and results indicated that they could be useful to the study of vascular biology. Nevertheless, both the models aim to reduce the use of experimental animals and to refine animal based-trials. In conclusion, the present research aims to be a small, but significant, contribution to the important and necessary field of study of alternative methods to animal experimentation.

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Giaretta, Elisa <1985&gt. "Effect of antioxidant supplementation on pig and horse gamete storage." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amsdottorato.unibo.it/6950/1/Giaretta_Elisa_tesi.pdf.

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According to recent studies, antioxidant supplementation on gamete processing and/or storage solutions improvesgamete quality parameters, after cooling or storage at sub zero temperature. The aim of the present study was to investigate the effects of antioxidant supplementation on pig and horse gamete storage. The first study aimed to determine the effects of resveratrol (RESV) on the apoptotic status of porcine oocytes vitrified by Cryotop method, evaluating phosphatidylserine (PS) exteriorization and caspases activation. RESV(2µM) was added during: IVM (A); 2 h post-warming incubation (B); vitrification/warming and 2 h post-warming incubation (C); all previous phases (D). The obtained data demonstrate that RESV supplementation in the various steps of IVM and vitrification/warming procedure can modulate the apoptotic process, improving the resistance of porcine oocytes to cryopreservation-induced damage. In the second work different concentrations of RESV (10, 20, 40, and 80µM) were added during liquid storage of stallion sperm for 24 hours at either 10°C or 4°C, under anaerobic conditions. Our findings demonstrate that RESV supplementation does not enhance sperm quality of stallion semen after 24 hours of storage. Moreover, the highest RESV concentrations tested (40 and 80µM) could damage sperm functional status, probably acting as pro-oxidant. Finally, in the third work other two antioxidants, ascorbic acid (AA) (100 µM) and glutathione (GSH) (5mM) were added on boar freezing and/or thawing solutions. In our study different sperm parameters were evaluated before freezing and at 30 and 240 minutes after thawing. Our results showed that GSH and AA significantly improved boar sperm cryotolerance, especially when supplemented together to both freezing and thawing media. This improvement was observed in sperm viability and acrosome integrity, sperm motility, and nucleoprotein structure. Although ROS levels were not much increased by freeze-thawing procedures, the addition of GSH and AA to both freezing and thawing extenders significantly decreased intracellular peroxide levels.

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Giaretta, Elisa <1985&gt. "Effect of antioxidant supplementation on pig and horse gamete storage." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amsdottorato.unibo.it/6950/.

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According to recent studies, antioxidant supplementation on gamete processing and/or storage solutions improvesgamete quality parameters, after cooling or storage at sub zero temperature. The aim of the present study was to investigate the effects of antioxidant supplementation on pig and horse gamete storage. The first study aimed to determine the effects of resveratrol (RESV) on the apoptotic status of porcine oocytes vitrified by Cryotop method, evaluating phosphatidylserine (PS) exteriorization and caspases activation. RESV(2µM) was added during: IVM (A); 2 h post-warming incubation (B); vitrification/warming and 2 h post-warming incubation (C); all previous phases (D). The obtained data demonstrate that RESV supplementation in the various steps of IVM and vitrification/warming procedure can modulate the apoptotic process, improving the resistance of porcine oocytes to cryopreservation-induced damage. In the second work different concentrations of RESV (10, 20, 40, and 80µM) were added during liquid storage of stallion sperm for 24 hours at either 10°C or 4°C, under anaerobic conditions. Our findings demonstrate that RESV supplementation does not enhance sperm quality of stallion semen after 24 hours of storage. Moreover, the highest RESV concentrations tested (40 and 80µM) could damage sperm functional status, probably acting as pro-oxidant. Finally, in the third work other two antioxidants, ascorbic acid (AA) (100 µM) and glutathione (GSH) (5mM) were added on boar freezing and/or thawing solutions. In our study different sperm parameters were evaluated before freezing and at 30 and 240 minutes after thawing. Our results showed that GSH and AA significantly improved boar sperm cryotolerance, especially when supplemented together to both freezing and thawing media. This improvement was observed in sperm viability and acrosome integrity, sperm motility, and nucleoprotein structure. Although ROS levels were not much increased by freeze-thawing procedures, the addition of GSH and AA to both freezing and thawing extenders significantly decreased intracellular peroxide levels.

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38

Mandelli, Michaela <1977&gt. "Studies on the reproductive Physiology of two critically endangered species of the North Adriatic Sea: Adriatic sturgeon (Acipenser naccarii) and European eel (Anguilla anguilla)." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2016. http://amsdottorato.unibo.it/7459/1/thesisdone.pdf.

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Reproductive physiology has many applications for a successful management of fish population in aquaculture. In particular the stock management of endangered species might take advantage of the knowledge concerning the reproductive cycle in order to improve the protocols and restore the population for restock activities. The Adriatic sturgeon (Acipenser naccarrii) and the European eel (Anguilla anguilla) are two endemic species of the North Adriatic Sea and are both listed in the the IUCN Red List of threatened species as “critically endangered”. Both the species nowadays rely on the experimental activities and new technologies that try to implement the farming conditions for an improvement of the population management for restocking purposes. In the present study the onset of puberty was investigated through plasma Testosterone analysis in A. naccarii. With reference to A. Anguilla the effect of photoperiod was assessed on the endocrine profiles during hormonal induction. Furthermore a new tank design has lead to spontaneous spawning that was then compared to the manual stripping protocol.

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Mandelli, Michaela <1977&gt. "Studies on the reproductive Physiology of two critically endangered species of the North Adriatic Sea: Adriatic sturgeon (Acipenser naccarii) and European eel (Anguilla anguilla)." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2016. http://amsdottorato.unibo.it/7459/.

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Reproductive physiology has many applications for a successful management of fish population in aquaculture. In particular the stock management of endangered species might take advantage of the knowledge concerning the reproductive cycle in order to improve the protocols and restore the population for restock activities. The Adriatic sturgeon (Acipenser naccarrii) and the European eel (Anguilla anguilla) are two endemic species of the North Adriatic Sea and are both listed in the the IUCN Red List of threatened species as “critically endangered”. Both the species nowadays rely on the experimental activities and new technologies that try to implement the farming conditions for an improvement of the population management for restocking purposes. In the present study the onset of puberty was investigated through plasma Testosterone analysis in A. naccarii. With reference to A. Anguilla the effect of photoperiod was assessed on the endocrine profiles during hormonal induction. Furthermore a new tank design has lead to spontaneous spawning that was then compared to the manual stripping protocol.

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40

Ventrella, Domenico <1987&gt. "The Piglet as Biomedical Model: Physiological Investigations, New Techniques and Future Applications." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2017. http://amsdottorato.unibo.it/7852/1/ventrella_domenico_tesi.pdf.

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By the analysis of the literature, it looks like the strive for new piglets’ models of disease have prevaricated the necessity for a deeper knowledge of the physiology of the animal. It is well known how difficult it is to interpret obtained data when poor to none reference standards are provided and how hard it is to apply techniques borrowed from other models, no matter how similar they can look. The aim of the present work was to collect knowledge and information regarding the piglets by studying its physiology and to validating new techniques. The experiments can be divided into two categories: physiological investigations and new techniques and future applications. Blood and Cerebrospinal fluid were analyzed in a quali-quantitative manner in piglets. The studies, thanks to the extensive statistical analyses and the high number of sampled population, provide with important reference intervals that will allow for better understanding of several metabolic processes. For the second category, technical experiments aimed to find easier and relatively pain free procedures for the collection of Cerebrospinal fluid and intrathecal administration in piglets, were performed. Operators’ skills often are a limiting factor for the feasibility of experimental protocols, and easier techniques are the best way to break down these walls. Moreover, when leading to lower mortality and higher welfare, those techniques allow for better results and higher ethical standards. The last experiment aimed to create a comprehensive map of CNS transduction upon intrathecal administration of Adeno-Associated Viral vector in piglets, and to evaluate their potential toxicity. The obtained result will help choosing the right serotype depending on the targeted cell population, thus avoiding preliminary studies reducing the number of enrolled animals. In conclusion, this thesis represents an additional step toward the standardization of the physiological piglet model and its refinement and reduction in experimental protocols.

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Borjini, Nozha <1988&gt. "Early Markers of Microglia Activation in Inflammatory Diseases." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2017. http://amsdottorato.unibo.it/7895/1/Borjini_Nozha_Tesi.pdf.

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The last two decades has witnessed many achievements in our understanding of the molecular-mechanisms underlying various neuroinflammatory-disorders. Microglia activation is thought to be a driving force of neurodegeneration that follows neuroinflammation in many neurological disorders, but confirmatory evidence is still elusive. In particular, the possible relationship between cause and consequence for microglia activation and pathological landmarks, such as neuronal demyelination and cell death in adult vs neonatal age is still disputed. In this thesis we tried to highlight the potential of early biomarkers for microglia-activation, using two rat models of diseases where microglia activation and neurodegeneration interact. In the paper included in chapter I, we performed a time-course investigation of neuroinflammation and demyelination biomarkers in the spinal cord, cerebrospinal fluid and blood in EAE induced in Dark-Agouti female rats compared with controls and adjuvant, focusing on the time-course between immunization and clinical-onset. We demonstrate that CSF1 was the first up-regulated protein at 1 DPI, in blood, cerebrospinal fluid and spinal cord. A treatment with GW2580, a selective CSF1R inhibitor, slowed the disease progression, significantly reduced the severity and prevented the relapse phase. Moreover, both pro-and ant-inflammatory cytokines were regulated starting from 8 DPI. In the manuscript included in chapter II, we investigated the effect of GW2580 on blood brain barrier disruption with the temporal evolution of EAE. We demonstrated that GW2580 treatment had a therapeutic effect in EAE rats, through reduction of BBB leakage by inhibiting activities of MMP-9 and consequent reduction of microglia activation, IgG-extravasation, and T-cell infiltration. In the manuscript included in chapter III, we investigated plasma and CSF-contents of inflammatory biomarkers after neonatal-HI on acute and chronic phases and their correlation with neurological disorders in rat model of HI. Our data revealed that several inflammatory modulators were most affected at the acute-phase and stabilized at the chronic-phase

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Barone, Francesca <1984&gt. "The Pig in Translational Medicine: Visual and Gastro-Intestinal Systems Applications." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2017. http://amsdottorato.unibo.it/8097/1/Barone_Francesca_tesi.pdf.

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Scientists are becoming every day more aware that a deep knowledge of an animal used in biomedical science can lead to more reliable data. The translational value of animal models is becoming an important topic of debate due to the high frequency of fail when animal experiments are replicate on humans. The first aim of this work was to better characterize the role of nutrition on the gut microbiota in the pig model. The second aim of my work was to investigate a chemically inducible retinal degeneration pig model. The results lead to a thorough understanding of pig gut microbiota and its modulation to nutritional changes, these understandings are fundamental for the interpretation of data when experiments are conducted on animals. The potential role in reduction and refinement is clear especially considering the pig as the emerging gold standard model for gastro intestinal research. The microencapsulation of iron was found to be protective against the side effects of iron dietary supplementation maintaining the healing effect, opening a new prospective on iron fortified foods. New tools have been established for the assessment of vision in a pig model. An easy visual evaluation behavioral test has been validated, giving to researchers and clinicians the opportunity to assess vision in a sentient pig model. Moreover the pattern electroretinogram (pERG) was recorded for the first time in pigs. The pERG is a sensitive indicator of dysfunction within the macular region and it reflects the integrity of the optics, photoreceptors, bipolar cells and retinal ganglion cells. Hopefully this work will contribute to the translational value of the pig model, which in my opinion is the future gold standard model for gastro-intestinal and vision systems and will completely replace the use of dogs and non-humane primates in biomedical science.

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Gadani, Beatrice <1989&gt. "Methods for improving boar and stallion semen quality." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2018. http://amsdottorato.unibo.it/8386/1/Gadani_Beatrice_tesi.pdf.

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Handling of semen is a very important tool for a successful artificial insemination in both pig and horse farming. One of the main problems during liquid and frozen semen storage is the loss of fertilizing potential mainly due to a sudden reduction of temperature, an excessive production of reactive oxygen species and alterations in the antioxidant defense systems. Supplementation of sperm preparation with different antioxidants gave interesting and promising results and, during recent years, use of plant antioxidants has been gaining the attention of several research groups. Moreover, an increase in the fertilising ability of boar sperm has been demonstrated after photo-stimulation. On these bases, the objective of the present thesis was to assess whether: 1) Resveratrol (RESV) or Epigallocatechin-3-gallate (EGCG) supplementation of thawing boar semen extender is effective in influencing sperm quality parameters and in vitro fertilization ability; 2) EGCG and green tea extract polyphenols improve stallion semen parameters during cooling at 4°C; 3) Photo-stimulation improves frozen-thawed boar semen quality. The results obtained demonstrate that the addition of RESV or EGCG, alone or in combination, to thawed boar semen positively affects in vitro penetration rate while no positive effects were registered in cooled stallion semen up to 48 h storage; as a consequence, the addition of these substances in cooling medium for stallion sperm storage is not useful. Therefore, it is evident that the effects of antioxidants may vary depending on species as well as on the way semen is processed for preservation (cooled vs frozen-thawed). Moreover, photo-stimulation may be considered as a potential tool to increase the cryotolerance of poor freezability ejaculates (PFE) but more researches on the mechanisms underlying the detrimental effects on sperm motility are required; they may allow to understand whether such increase in cryotolerance has a significant impact upon the fertilizing ability of PFE.

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Petrulli, Carmen Adele <1982&gt. "Custodi erranti: il mio viaggio tra cani, pecore e lupi. Valutazione comportamentale di Cani da Pastore Abruzzesi utilizzati come strumento di conservazione della biodiversità e mitigazione del conflitto zootecnico in Toscana." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2018. http://amsdottorato.unibo.it/8713/13/CUSTODI%20ERRANTI%20IL%20MIO%20VIAGGIO%20TRA%20CANI%2C%20PECORE%20E%20LUPI.pdf.

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Il lavoro da me svolto da dicembre 2014 a gennaio 2016, ha avuto lo scopo di valutare sperimentalmente nei cani da guardianìa (LGD), l’attitudine alla protezione delle greggi e l’efficacia di tale metodo di prevenzione nei confronti dei danni da predazione al bestiame nelle aziende oggetto di studio.
This study took place from December 2014 to January 2016. The aim of the study was to evaluate in Abruzzi Sheepdogs their aptitude towards livestock protection and the effictiveness of this preventive method towards predator's damage to the sheeps of the farms studied.

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Tubon, Usca Irvin Ricardo <1988&gt. "Anti-inflammatory activity of plant extracts: an effective in vitro model of study based on primary culture of porcine endothelial cells." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2019. http://amsdottorato.unibo.it/8935/1/Tubon_Irvin_Tesi.pdf.

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Inflammation is a protective response exerted by the organism to eliminate an injurious stimulus. Endothelial cells play a fundamental role in different physiological processes since, changing their phenotype, modulate the complement and coagulation cascades, inflammation and innate and adaptive immunity. Medicinal plants have been the main remedy to treat different ailments and their study has permitted the development of many drugs. The aim of the present research was to study the phytochemical composition and the anti-inflammatory activity of four medicinal plants (Cucumis sativus L, Boswellia serrata Roxb. Colebr, Salvia sagittata Ruiz & Pav and Clinopodium tomentosum Kunth), traditionally used to treat different diseases, on primary culture of porcine aortic endothelial cells (pAECs) stimulated with Lipopolysaccharide (LPS). This approach is relevant in terms of screening possible positive activities of medicinal plants in relation to the Replacement and Reduction principles in agreement with the European Directive 2010/63/EU on the protection of animals used for scientific purposes and its implementation in Italy through “Decreto Legislativo“ n. 26/2014. Different extraction processes and techniques have been employed to verify their phytochemical and biological effects (HPLC, RT-PCR, Western blot). The phytochemical analysis revealed the presence of different bioactive compounds (11-keto-β-boswellic acid and β-boswellic acid in Boswellia serrata extracts, different phenolic acids and flavonoids with rosmarinic acid as the most abundant component in Salvia sagittata and Clinopodium tomentosum extracts). In the biological analysis, Cucumis sativus, Boswellia serrata, Salvia sagittata and Clinopodium tomentosum reduced the LPS-induced cytotoxicity with the decrement of different inflammatory cytokines and increment of protective molecules and anti-inflammatory cytokines. In conclusion, our results have clearly demonstrated that the medicinal plants studied have attenuated lipopolysaccharide induced inflammatory response in endothelial cells. Moreover the studies confirm the relevance of primary culture of pAECs as an interesting screening model to explore the huge variety of Ecuadorian medicinal plants.

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Postiglione, Gabriella <1986&gt. "Practical Management of the Endangered African Wild Dog (Lycaon Pictus): Implementation of Research Methodologies, Management Tools and Validation of Non-Invasive Endocrinology Applications." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2019. http://amsdottorato.unibo.it/9092/1/Postiglione%20PhD%20thesis%20-%2029.09.19%20FRONTESPIZIO%20COMPLETA.pdf.

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The porpoise of this study was to implement research methodologies and assess the effectiveness and impact of management tools to promote best practices for the long term conservation of the endangered African wild dog (Lycaon pictus). Different methods were included in the project framework to investigate and expand the applicability of these methodologies to free-ranging African wild dogs in the southern African region: ethology, behavioural endocrinology and ecology field methodologies were tested and implemented. Additionally, research was performed to test the effectiveness and implication of a contraceptive implant (Suprenolin) as a management tool for the species of a subpopulation hosted in fenced areas. Attention was especially given to social structure and survival of treated packs. This research provides useful tools and advances the applicability of these methods for field studies, standardizing and improving research instruments in the field of conservation biology and behavioural endocrinology. Results reported here provide effective methodologies to expand the applicability of non-invasive endocrine assessment to previously prohibited fields, and validation of sampling methods for faecal hormone analysis. The final aim was to fill a knowledge gap on behaviours of the species and provide a common ground for future researchers to apply non-invasive methods to this species research and to test the effectiveness of the contraception on a managed metapopulation.

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Elmi, Alberto <1988&gt. "Essential oils and swine reproduction: new frontiers in antibiotic replacement in seminal doses." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2020. http://amsdottorato.unibo.it/9207/1/TESI_ELMI_repository.pdf.

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The actual scenario regarding antibiotic resistance is extremely worrying and the scientific community invest time and efforts in the search for solutions. Essential oils (EOs) are products of the secondary metabolism of aromatic plants and are complex mixtures of several compounds. EOs show a wide variety of biological activities, that may suggest interesting applications in the reproduction field, as cryo-preservatives or substitutes for antibiotics, but studies regarding their effects on spermatozoa are lacking. The present research aimed at evaluating the opportunity to use essential oils in refrigerated swine seminal doses as a potential alternative to antibiotics. The preliminary step was about the study of swine seminal parameters in farm setting using advanced statistical methods, to better predict in vivo fertility on the basis of semen morpho-functional evaluations. In the first step some specific EOs were selected and the cytotoxic effects on swine spermatozoa were evaluated to identify non-spermicidal concentrations. The lower concentrations of Rosmarinus officinalis and Melaleuca alternifolia EOs not impacted the semen morpho-functinal parameters. The last step was the evaluation of antibacterial effects of the previously tested concentrations on the liquid phase of swine refrigerated seminal doses. In order to obtain standardized results, an in vitro model with the addition of E. coli was set up. Analyses, performed at 24 and 120 hours, included optical density evaluation, bacterial DNA quantification by qPCR, and colony count. The results demonstrated that both EOs, at a concentration of 0.4 mg/ml, are capable of delivering similar effects to ampicillin. At the lower concentration, M. alternifolia EO seemed more effective when compared to R. officinalis. In conclusion, this project showed the potential of some EOs in the field of swine artificial insemination, but also highlighted how it is necessary, before claiming natural compounds as future “miracle-workers”, to investigate their safety and mechanisms of action.

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Manservisi, Fabiana <1981&gt. "Reproductive and developmental toxicity study using Sprague-Dawley rats exposed under various calendars to the weedkiller Glyphosate and commercial formulations Glyphosate-based." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2021. http://amsdottorato.unibo.it/9579/1/Manservisi_Fabiana_tesi.pdf.

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Glyphosate-based herbicides (GBHs) are the most globally used herbicides raising the risk of environmental exposition. Carcinogenic effects are only one component of the multiple adverse health effects of Glyphosate and GBHs that have been reported. Questions related to hazards and corresponding risks identified in relation to endocrine disrupting effects are rising. The present study investigated the possible reproductive/developmental toxicity of GBHs administered to male and female Sprague-Dawley rats under various calendar of treatment. Assessments included maternal and reproductive outcome of F0 and F1 dams exposed to GBHs throughout pregnancy and lactation and developmental landmarks and sexual characteristics of offspring. The study was designed in two stages. In the first stage Glyphosate, or its commercial formulation Roundup Bioflow, was administered to rats at the dose of 1.75 mg/kg bw/day (Glyphosate US Acceptable Daily Intake) from the prenatal period until adulthood. In the second stage, multiple toxicological parameters were simultaneously assessed, including multigeneration reproductive/developmental toxicity of Glyphosate and two GBHs (Roundup Bioflow and Ranger Pro). Man-equivalent doses, beginning from 0.5 mg/kg bw/day (ADI Europe) up to 50 mg/kg bw/day (NOAEL Glyphosate), were administered to male and female rats, covering specific windows of biological susceptibility. The results of stage 1 and preliminary data from stage 2 experiments characterize GBHs as probable endocrine disruptors as suggested by: 1) androgen-like effects of Roundup Bioflow, including a significant increase of anogenital distances in both males and females, delay of first estrous and increased testosterone in females; 2) slight puberty onset anticipation in the high dose of Ranger Pro group, observed in the F1 generation treated from in utero life until adulthood; 3) a delayed balano-preputial separation achievement in the high dose of Ranger Pro-treated males exposed only during the peri-pubertal period, indicating a direct and specific effect of GBHs depending on the timing of exposure.

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Nerozzi, Chiara <1992&gt. "Effects of glyphosate and Roundup upon mammalian gametes." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2021. http://amsdottorato.unibo.it/9686/1/Tesi%20Dottorato%20Dott.ssa%20Chiara%20Nerozzi.pdf.

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The wide use of glyphosate-based herbicides (GBHs) has become a controversial issue due to the potential harmful effects on human health. Commercial formulations, among which Roundup is the most famous one, contain a number of adjuvants inside; most of these are patented and not publicly known, therefore, they can act differently from glyphosate alone and might strengthen its toxic effect. Our study is focused on GBHs reproductive toxicity with a special regard to glyphosate and Roundup impact on male and female mammalian gametes after exposure to concentrations ranging from the one recommended for agricultural use (0.1% Roundup, containing 360 µg/mL glyphosate) to 70-fold lower or more. Sperm quality analysis, either on boar and stallion, showed that Roundup has much more detrimental impact than glyphosate at equivalent concentrations on spermatozoa function and survival. Basing on our results, the toxic effect of these pesticides on spermatozoa may be linked to an impairment in mitochondrial activity and a subsequent decrease in ATP production and/or alterations in the redox balance, which impact cell motility and plasma membrane stability. Moreover, a different species sensitivity to GBHs may exists as high doses of glyphosate affected sperm quality only in boar and not in stallion; furthermore, Roundup had deleterious effects at lower doses in the first compared to the latter. With regard to female gametes, we found that glyphosate and Roundup exposure during IVM detrimentally affect the subsequent developmental ability of swine embryos, providing further evidence of their potential toxic effect on female reproductive system. In addition, Roundup altered steroidogenesis and increased oocyte ROS levels. Therefore, according to our results, we can conclude that GBHs exert a negative impact on both male and female gametes and that Roundup adjuvants enhance glyphosate toxic effects and/or are biologically active in their side-effect.

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Fuentes, Rubio María. "Estudios sobre alfa-amilasa en saliva como marcador de estrés en la especie humana y otras especies animales= Studies on salivary alpha-amylase as a maker of stress in human and different animal species." Doctoral thesis, Universidad de Murcia, 2014. http://hdl.handle.net/10803/277100.

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Tesis por compendio de publicaciones
OBJETIVOS 1. Evaluar el uso de la alfa amilasa salivar como marcador de estrés usando técnicas tradicionales (ensayos enzimáticos) en humana, usando como modelo experimental a estudiantes de veterinaria que tenían que realizar una presentación oral de un trabajo en lengua extranjera. 2. Evaluar el uso de la alfa amilasa salivar como marcador de estrés usando técnicas tradicionales (ensayos enzimáticos) en cerdos. 3. Implementar nuevos enfoques para la medición de la alfa-amilasa en muestras de saliva de varias especies (incluidos los seres humanos) mediante la cuantificación directa de la cantidad de la alfa-amilasa salival en lugar de mediante la medición de su actividad 4. Detectar e identificar nuevos biomarcadores de estrés en la saliva mediante técnicas de proteómica. Una técnica avanzada que permite la separación e identificación de múltiples proteínas. METODOLOGÍA Esta tesis doctoral es un compendio de los siguientes artículos publicados: 1) Assessment of Stress Associated with an Oral Public Speech in Veterinary Students by Salivary Biomarkers. Tecles F, Fuentes-Rubio M, Tvarijonaviciute A, Martínez-Subiela S, Fatjó J, Cerón JJ. (2014) J Vet Med Educ. Jan 21:1-7. [Epub ahead of print]. 2) Validation of an automated method for salivary alpha-amylase measurements in pigs (Sus scrofa domesticus) and its application as a stress biomarker. Fuentes M, Tecles F, Gutiérrez A, Otal J, Martínez-Subiela S, Cerón JJ. (2011) J Vet Diagn Invest. Mar;23(2):282-7. 3) Porcine salivary analysis by two-dimensional gel electrophoresis in three acute stress models: a pilot study. Fuentes-Rubio M, Cerón J, de Torre C, Escribano D, Gutiérrez AM, Tecles F. (2014) Can J Vet Res. Apr;78(2):127-32. En el primer artículo, se midió el cortisol salival y la actividad alfa-amilasa para evaluar el estrés asociado con una presentación oral pública de los estudiantes de Patología Clínica Veterinaria. Se recogieron muestras de saliva antes y directamente después de la presentación oral de 5 minutos y a los 20 y 35 minutos después del comienzo de la misma. El cortisol alcanzó su punto máximo 20 minutos después del comienzo de la presentación, mientras que la alfa-amilasa salival alcanzó su máximo justo al final del discurso. No existió relación entre estos cambios y el nivel de estrés que se indica por el estudiante en un cuestionario, ni con el sexo del estudiante, o la calidad de la presentación. El objetivo del segundo estudio fue validar un método espectrofotométrico automatizado para la medición de alfa-amilasa salival en cerdos y evaluar su posible aplicación como biomarcador de estrés no invasivo . La validación analítica incluyó la precisión intra- e inter- ensayo, linealidad bajo dilución, y límite de detección. Además, para estudiar el posible uso de alfa- amilasa salival como marcador de estrés, 12 cerdos en crecimiento de 3-4 meses de edad fueron sometidos a estrés por inmovilización durante al menos 1 min, obteniéndose muestras de saliva en diferentes puntos de tiempo. Los resultados de la validación analítica indicaron que el método era preciso y capaz de medir la alfa-amilasa de una manera lineal. Los resultados obtenidos en la prueba de esfuerzo mostraron un aumento significativo en la actividad salival alfa amilasa. El propósito del artículo 3 fue estudiar los cambios en el proteoma salival de cerdos sanos en situaciones de estrés para identificar cualquier posible nuevo biomarcador salival de estrés. Tres grupos de animales fueron sometidos a 3 modelos de estrés: restricción seguido de muestreo simulado de sangre de la vena cava; transporte breve por carretera; restricción de movimiento en una jaula digestibilidad. La saliva se obtuvo de cada animal antes, 15 y 30 min después de la inducción de estrés. Las muestras de los animales que mostraron el mayor incremento en la concentración de cortisol salivar se agruparon y se realizó una electroforesis en geles bidimensionales. Se tiñeron con Coomassie Brilliant Blue R-250 para la detección de spots y seleccionarlos para su identificación mediante espectrometría de masas. Los análisis estadísticos demostraron que 2 proteínas tenían diferencias significativas en la expresión antes y después de la inducción de estrés y fueron identificadas como fragmentos de albúmina y proteína de unión odorizante. CONCLUSIONES 1. La actividad enzimática de la alfa amilasa salivar puede ser usada como biomarcador de estrés en estudiantes de Veterinaria tras someter a los individuos a una presentación pública oral. 2. La actividad enzimática de la alfa amilasa salivar puede ser usada como biomarcador de estrés en cerdos tras someter a los individuos a un protocolo de restricción de movimientos. 3. La concentración de alfa amilasa en saliva de humanos, cerdos, caballos y ovejas puede ser cuantificada mediante un ensayo de inmunofluorometría a tiempo resuelto. Este ensayo tiene una variación inter-individual baja y es más sensible que los ensayos enzimáticos para detectar estrés. 4. El proteoma salivar de los cerdos ha revelado diferencias entre animales en condiciones basales y bajo condiciones de estrés. Esto se reveló como una herramienta útil para buscar posibles nuevos biomarcadores de estrés in estos animales tales como la proteína de unión odorizante.
OBJECTIVES 1. To evaluate the use of salivary alpha-amylase as a marker of stress using traditional techniques (enzymatic assays) in human using a model in which veterinary students had to make an oral presentation in a foreing language. 2. To evaluate the use of salivary alpha-amylase as a marker of stress using traditional techniques (enzymatic assays) in pigs. 3. To implement new approaches for measuring alpha-amylase in saliva samples of several species (including human beings) by direct quantification of the amount of the salivary alpha-amylase instead of by measuring its activity (Article 3, Article 4, Article 5, Article 6). 4. To detect and identify new biomarkers of stress in saliva by proteomics. An advanced technique that allows separation and identification of multiple proteins. METHODOLOGY This PhD thesis is a compilation of the following published papers: 1) Assessment of Stress Associated with an Oral Public Speech in Veterinary Students by Salivary Biomarkers. Tecles F, Fuentes-Rubio M, Tvarijonaviciute A, Martínez-Subiela S, Fatjó J, Cerón JJ. (2014) J Vet Med Educ. Jan 21:1-7. [Epub ahead of print]. 2) Validation of an automated method for salivary alpha-amylase measurements in pigs (Sus scrofa domesticus) and its application as a stress biomarker. Fuentes M, Tecles F, Gutiérrez A, Otal J, Martínez-Subiela S, Cerón JJ. (2011) J Vet Diagn Invest. Mar;23(2):282-7. 3) Porcine salivary analysis by two-dimensional gel electrophoresis in three acute stress models: a pilot study. Fuentes-Rubio M, Cerón J, de Torre C, Escribano D, Gutiérrez AM, Tecles F. (2014) Can J Vet Res. Apr;78(2):127-32. In the first article, salivary cortisol and alpha-amylase activity were measured to assess stress associated with a public oral presentation in Veterinary Clinical Pathology students. Stimulated saliva samples were collected before and directly after a 5-minute oral presentation and at 20 and 35 minutes after the beginning of the presentation. Cortisol peaked 20 minutes after the beginning of the presentation, whereas salivary alpha-amylase (sAA) peaked at 5 minutes, just at the end of the speech. These changes were not related to the level of stress that was indicated by the student in a questionnaire, the student’s sex, or the quality of the presentation. The aim of the second study was to validate an automated spectrophotometric method for salivary alpha-amylase measurement in pigs and evaluate its possible application as a noninvasive stress biomarker. The analytical validation included intra- and interassay precision, linearity under dilution, and limit of detection. In addition, to study the possible use of salivary alpha-amylase as a possible stress marker, 12 crossbred growing pigs of 3-4 months of age were subjected to restraint stress by a nasal snare for at least 1 min, and saliva samples were obtained at different time points. The results of analytical validation indicated that the method was precise and able to measure alpha-amylase in a linear manner. The results obtained in the stress test showed a significant increase in salivary alpha-amylase activity. The purpose of the research of article 3 was to study changes in the salivary proteome of healthy pigs in stressful situations to identify any potential new salivary biomarker of stress. Three groups of animals were subjected to 3 stress models: snaring restraint followed by simulated sampling of vena cava blood; brief transport by road; and restriction of movement in a digestibility cage. Saliva was obtained from each animal before and 15 and 30 min after the induction of stress. The samples from the animals that showed the greatest increase in salivary cortisol concentration were pooled and run on 2-dimensional gels. Coomassie Brilliant Blue R-250 was used for spot detection and mass spectrometry for spot identification. Statistical analyses showed that 2 proteins had significant differences in expression before and after the induction of stress. These proteins were identified as odorant-binding protein and fragments of albumin. CONCLUSIONS 1. The enzymatic activity of salivary alpha-amylase can be used as biomarker of stress in Veterinary students after subjecting the individuals to a public oral presentation. 2. The enzymatic activity of salivary alpha-amylase can be used as biomarker of stress in pigs after subjecting the individuals to restraint of movement protocol. 3. Alpha-amylase concentration in saliva of humans, pigs, horses and sheep can be quantified by TR-IFMA This assay has a lower inter-individual variation and is more sensitive than enzymatic assays to detect stress. 4. Salivary proteome of pigs has revealed differences between animals in basal status and under stress conditions. It was shown as a useful tool to search possible new salivary biomarkers of stress in these animals such as odorant-binding protein.

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