Please read the abstract in the dissertation. / Dissertation (MSc Agric)--University of Pretoria, 2013. / gm2014 / Animal and Wildlife Sciences / unrestricted

Consumption of endophyte-infected tall fescue results in a syndrome of negatively altered physiological systems, collectively known as fescue toxicosis. Another challenge to endophyte-infected tall fescue -based beef cattle operations is that the soils often are selenium (Se) poor, necessitating the need to provide supplemental Se. To test the general hypothesis that different forms of supplemental Se would ameliorate the negative effects of fescue toxicosis, predominately-Angus steers (BW = 183 ± 34 kg) were randomly selected from herds of fall-calving cows grazing an endophyte-infected tall fescue pasture and consuming vitamin-mineral mixes that contained 35 ppm Se as sodium selenite (ISe), SELPLEX (OSe), or an 1:1 blend of ISe and OSe (MIX). Steers were commonly weaned and depleted of Se for 98 d. Steers were assigned (n = 8 per treatment) to the same Se-form treatments upon which they were raised and subjected to summer-long common grazing of an endophyte-infected tall fescue pasture (0.51 ppm ergot alkaloids: ergovaline plus ergovalinine; 10.1 ha). Selenium treatments were administered by daily top-dressing 85 g of vitamin-mineral mix onto 0.23 kg soyhulls, using in-pasture Calan gates. The first project objective was to determine the effect of forms of supplemental Se on whole blood Se, serum prolactin, liver glutamine synthetase (GS) activity, carcass parameters, and growth performance (Experiment 1). In Experiment 1, whole blood Se increased for all treatments from day 0 to 22 and then did not change. Across periods, MIX and OSe steers had greater whole blood Se than ISe steer. Compared to ISe steers, MIX and OSe steers had more serum prolactin. Liver GS mRNA, protein content, and activity were greater in MIX and OSe steers than ISe steers. However, the ADG and carcass parameters were not affected by Se treatments. The second project objective was to determine the effect of forms of supplemental Se on serum clinical parameters of Experiment 1 steers (Experiment 2). In Experiment 2, across periods, MIX steers had more serum albumin than OSe, and ISe steers, respectively. Serum alkaline phosphatase (ALP) activity was greater in MIX and OSe steers. In addition, blood urea nitrogen (BUN), serum sodium, phosphorus, and magnesium concentration were affected by Se treatments. Partial correlation analysis revealed that serum albumin, BUN, and ALP activity were correlated with whole blood Se concentration. The third project objective was to evaluate the hepatic transcriptome profiles of Experiment 1 steers using microarray and targeted RT-PCR analyses (Experiment 3). In Experiment 3, bioinformatic analysis of microarray data indicated that hepatic glutamate/glutamine, proline, arginine, and citrulline metabolism was affected by different forms of supplemental Se. The mRNA expression of critical proteins involved in glutamate/glutamine (GLS2, GLUD1, GLUL), proline (PYCR1, ALDH18A1), and urea (ARG1, ARG2, OAT, NAGS, OTC, ORNT1) metabolism were differentially expressed by Se treatments. Collectively, we conclude that consumption of 3 mg Se/d as OSe or MIX forms of Se in vitamin-mineral mixes 1) increased whole blood Se content, an indicator of greater whole-body Se assimilation; 2) increased serum prolactin, albumin, and ALP, the reduction of which are hallmarks of fescue toxicosis; and 3) altered hepatic nitrogen metabolism, as indicated by changes in key enzymes of glutamate/glutamine, proline, and urea metabolism. However, 4) these positive effects on metabolic parameters were not accompanied by increased growth performance.

Aktuell gibt es für Selen (Se) keine Bedarfsempfehlungen für das Damwild (Dama dama) in Gehegehaltung. Diese Studie soll die typische Se-Aufnahme bei Gehegehaltung von Damwild ermitteln und klären, welche Parameter sich eignen, um die Se-Versorgung des Damwildes zu reflektieren. Dazu wurden 19 Damhirsche in zwei Gruppen unter identischen Bedingungen gehalten. Die Tiere ernährten sich von dem natürlichen Grasaufwuchs und Mischfutter (0,15 mg/kg TS bzw. 1,07 - 1,91 mg/kg TS). In Blut, Plasma und die Organen wurden der Se-Gehalt, die Aktivität der Se-abhängigen Glutathionperoxidase (GPx) sowie teilweise die Gesamt-GPx-Aktivität (gesGPx), die Aktivität der Glutathion-S-Transferase (GST) und die Expression verschiedener GPx analysiert. Durch die Se-Supplementierung wurden ein signifikanter Anstieg des Plasma-Se in der Versuchsgruppe und ein moderater Unterschied der Vollblut-Se-Konzentration sowie der Vollblut-GPx-Aktivität zwischen der Kontroll- und der Versuchsgruppe beobachtet (p = 0,08). Außerdem wurde in allen Organen der Versuchsgruppe ein höherer Se-Gehalt als in der Kontrollgruppe festgestellt. In der Hierarchie der untersuchten Organe ist die Niere am höchsten angeordnet, absteigend folgen der Herz- und Skelettmuskel, die Milz und die Leber. Eine Se-Aufnahme von 0,05 - 0,08 mg/kg TS führt beim Damwild nicht zur Ausprägung von Se-Mangelsymptomen und stellt daher eine ausreichende Se-Versorgung dar. Die Empfehlungen zur Se-Versorgung für Damwild sind somit nicht von Hauswiederkäuern zu übernehmen. Im Plasma und im Vollblut scheinen Se-Konzentrationen von 28 - 64 µg/l und 81 - 200 µg/l für eine ausreichende Se-Versorgung zu sprechen, in der Leber Se-Konzentrationen von 270 - 663 µg/kg TS.

Aktuell gibt es für Selen (Se) keine Bedarfsempfehlungen für das Damwild (Dama dama) in Gehegehaltung. Diese Studie soll die typische Se-Aufnahme bei Gehegehaltung von Damwild ermitteln und klären, welche Parameter sich eignen, um die Se-Versorgung des Damwildes zu reflektieren. Dazu wurden 19 Damhirsche in zwei Gruppen unter identischen Bedingungen gehalten. Die Tiere ernährten sich von dem natürlichen Grasaufwuchs und Mischfutter (0,15 mg/kg TS bzw. 1,07 - 1,91 mg/kg TS). In Blut, Plasma und die Organen wurden der Se-Gehalt, die Aktivität der Se-abhängigen Glutathionperoxidase (GPx) sowie teilweise die Gesamt-GPx-Aktivität (gesGPx), die Aktivität der Glutathion-S-Transferase (GST) und die Expression verschiedener GPx analysiert. Durch die Se-Supplementierung wurden ein signifikanter Anstieg des Plasma-Se in der Versuchsgruppe und ein moderater Unterschied der Vollblut-Se-Konzentration sowie der Vollblut-GPx-Aktivität zwischen der Kontroll- und der Versuchsgruppe beobachtet (p = 0,08). Außerdem wurde in allen Organen der Versuchsgruppe ein höherer Se-Gehalt als in der Kontrollgruppe festgestellt. In der Hierarchie der untersuchten Organe ist die Niere am höchsten angeordnet, absteigend folgen der Herz- und Skelettmuskel, die Milz und die Leber. Eine Se-Aufnahme von 0,05 - 0,08 mg/kg TS führt beim Damwild nicht zur Ausprägung von Se-Mangelsymptomen und stellt daher eine ausreichende Se-Versorgung dar. Die Empfehlungen zur Se-Versorgung für Damwild sind somit nicht von Hauswiederkäuern zu übernehmen. Im Plasma und im Vollblut scheinen Se-Konzentrationen von 28 - 64 µg/l und 81 - 200 µg/l für eine ausreichende Se-Versorgung zu sprechen, in der Leber Se-Konzentrationen von 270 - 663 µg/kg TS.:Inhaltsverzeichnis Inhaltsverzeichnis I Abbildungsverzeichnis V Tabellenverzeichnis VI Verzeichnis der Anhangstabellen VIII Abkürzungsverzeichnis IX 1. Einleitung 1 2. Literaturübersicht 2 2.1 Se als chemisches Element 2 2.2 Geschichte des Se und seiner Proteine 3 2.3 Se-Gehalte in Boden, Pflanzen, Nahrungs- und Futtermitteln 4 2.3.1 Se-Gehalte im Boden 4 2.3.2 Se-Gehalte in Pflanzen 5 2.3.3 Se-Gehalte in Nahrungsmitteln 7 2.3.4 Se-Gehalte in Futtermitteln 8 2.4 Se im Stoffwechsel 9 2.4.1 Resorption 9 2.4.2 Transport, Metabolismus und Speicherung 10 2.4.2.1 Transport 10 2.4.2.3 Speicherung 11 2.4.3 Versorgung über Plazenta und Milch 12 2.4.4 Exkretion 14 2.5 Biologische Funktionen des Se 16 2.5.1 SeP 16 2.5.2 Funktionen 18 2.5.2.1 Spezielle Funktionen der GPx 18 2.5.2.2 Weitere Funktionen der Selenoenzyme 20 2.6 Damwild (Dama dama) 21 2.6.1 Systematische und historische Einordnung des Damwildes 21 2.6.2 Physiologie und Ernährung des Damwildes 22 2.6.3 Se-Status bei Cerviden 23 2.7 Se-Bedarf 24 2.8 Se- und Enzymwerte im Organismus 25 2.8.1 Se-Gehalte im Blut 25 2.8.2 Se-Gehalte in verschiedenen Organen 28 2.9 Se und Erkrankungen 30 2.9.1 Se-Mangel assoziierte Erkrankungen 30 2.9.2 Se-Toxizität 31 2.9.2.1 Die akute Se-Intoxikation 32 2.9.2.2 Die subakute Se-Intoxikation 33 2.9.2.3 Die chronische Se-Intoxikation 33 3 Tiere, Material und Methoden 35 3.1 Versuchsziel 35 3.2 Tiere 35 3.3 Haltung 35 3.4 Fütterung und Supplementierung 35 3.5 Versuchsablauf 37 3.6 Probenentnahmen 38 3.6.1 Blutproben 38 3.6.2 Organ- und Gewebeproben 38 3.6.3 Wiegen 40 3.6.4 Futterproben 41 3.7 Versuchsparameter 43 3.8 Analytische Methoden 43 3.8.1 Futteranalyse 43 3.8.1.1 TS 43 3.8.1.2 Rohasche (Ra) 44 3.8.1.3 Organische Substanz (oS) 44 3.8.1.4 Rohprotein (Rp) 44 3.8.1.5 Rohfett (Rfe) 44 3.8.1.6 Rohfaser (Rfa) 44 3.8.1.7 N-freie Extraktstoffe (NfE) 45 3.8.1.8 Spurenelemente: Se, Cu, Zn 45 3.8.2 Vollblut-, Plasma-, Organ- und Gewebeanalyse 45 3.8.2.1 Histologie der Skelettmuskulatur 45 3.8.2.2 Se-Gehalt 46 3.8.2.3 TS-Gehalt 47 3.8.2.4 GPx-Aktivität 47 3.8.2.5 Proteingehalt 48 3.8.2.6 Hämoglobingehalt 49 3.8.2.7 GPx-mRNA-Expression 49 3.8.2.8 α -Glutathion-S-Transferase-Aktivität (GST) 53 3.9 Statistische Auswertung 54 4. Ergebnisse 55 4.1 KM der Tiere und Gewichte der Schlachtkörperhälften 55 4.1.1 KM der Tiere zu Versuchsbeginn 55 4.1.2 Gewichte der Schlachtkörperhälften zu Versuchende 55 4.2 Histologie der Skelettmuskulatur 56 4.3 Se-Gehalte in Plasma, Vollblut und Organen 58 4.3.1 Se-Gehalte in Plasma und Vollblut 58 4.3.2 Se-Gehalte in verschiedenen Organen 59 4.4 Se-abhängige und -unabhängige Enzyme 60 4.4.1 GPx-Aktivitäten in Plasma und Vollblut 60 4.4.2 GPx-Aktivität in verschiedenen Organen 62 4.4.3 GPx-mRNA-Expression 63 4.4.4 α-GST-Aktivität 64 5. Diskussion 67 5.1 Kritik der Methoden 67 5.1.1 Se-Supplementierung der Tiere 67 5.1.1.1 Futteraufnahme 67 5.1.1.2 Höhe der Se-Supplementierung 67 5.1.1.3 Dauer der Se-Supplementierung 68 5.1.1.4 Art der Se-Supplementierung 68 5.1.2 Probengewinnung 69 5.1.3 Untersuchungsparameter 69 5.1.4 Vitamin E 69 5.2 Diskussion der Versuchsergebnisse 70 5.2.1 Einschätzung der Se-Versorgung vor Se-Supplementierung 70 5.2.2 Einschätzung der Se-Versorgung nach unterschiedlicher Se-Supplementierung 72 6. Zusammenfassung 81 7. Summary 83 8. Literaturverzeichnis 85 9. Anhang……………………………………………………………………..110 Danksagung 117

A formulação de dietas para ruminantes pode alterar o perfil de ácidos do leite, potencializando o valor nutritivo e nutracêutico do mesmo. Estas alterações podem, no entanto, aumentar a susceptibilidade do leite à oxidação, perdendo qualidade em sabor e tempo de prateleira. O objetivo deste trabalho consistiu em avaliar o efeito do fornecimento de dieta com alto teor de soja integral extrusada, associada ou não à suplementação com selênio orgânico, sobre a produção, a composição, a concentração de CLA, o perfil de ácidos graxos e a estabilidade oxidativa do leite. Vinte e quatro vacas holandesas foram agrupadas em três blocos, de acordo com o número de lactações e alimentadas com dieta total contendo 56% de volumoso e 44% de concentrado. Os animais foram distribuídos aleatoriamente nos seguintes tratamentos: A) dieta controle, B) dieta contendo 21% de soja extrusada e C) dieta contendo 21% de soja extrusada + 5 mg de selênio orgânico. As dietas foram balanceadas para obtenção dos mesmos níveis de energia e proteína. O fornecimento de soja extrusada não afetou o consumo de matéria seca, o peso vivo dos animais e a produção de leite. As vacas alimentadas com soja extrusada produziram leite com menor (P<0,01) porcentagem de gordura, menor (P<0,01) concentração de ácidos graxos de cadeias curta e média e maior (P<0,01) concentração de ácidos graxos de cadeia longa, quando comparadas às vacas alimentadas com a dieta controle. Com relação ao teor de saturação, produziram leite com menor (P<0,01) concentração de ácidos graxos saturados e maior (P<0,01) concentração de ácidos graxos poliinsaturados e de ácido linoléico conjugado (CLA). A concentração de colesterol total foi maior (P<0,01) no soro sanguíneo das vacas alimentadas com soja extrusada, quando comparado ao controle. A suplementação com selênio aumentou (P<0,05) significativamente a concentração do mineral no leite e retardou a oxidação do mesmo, avaliada pelo índice de peróxido (P<0,01). O fornecimento da dieta contendo 21% de soja alterou o perfil de ácidos graxos do leite, diminuindo a concentração de ácidos graxos saturados e aumentando a concentração de ácidos graxos poliinsaturados e ácido linoléico conjugado, no entanto, produziu um leite mais susceptível à oxidação. O enriquecimento do leite com selênio conseguiu minimizar este efeito influenciando positivamente a estabilidade do leite. / The manipulation of ruminant diet can modify the fatty acid profiles increasing its nutritive and therapeutic value. However, this change in fatty acid composition can increase milk oxidation and can affect its flavor and shelf life. The objective of this study was to evaluate the effect of extruded soybean diet and organic selenium supplementation on milk composition, CLA content, fatty acid profiles and oxidative stability of milk. Twenty four Holstein cows were grouped into three blocks according to number of parturition and were fed diets containing 56% forage and 44% concentrate. The animals were allocated to three treatments: A) control diet, B) 21% extruded soybean diet and C) 21% extruded soybean diet + 5 mg of organic selenium. The diets were formulated to provide the same energy and protein levels. The extruded soybean did not affect the dry matter intake, body weight and milk production. The cows fed extruded soybean diet produced milk with lower (P<0,01) concentration of fat, lower (P<0,01) concentration of short and medium chain fatty acids and higher (P<0,01) concentration of long chain fatty acids when compared to control group. Regarding to saturation level, those animals produced milk with lower (P<0,01) concentration of saturated fatty acids and higher (P<0,01) concentration of polyunsaturated fatty acids and CLA content. The serum concentration of cholesterol was higher (P<0,01) for treatments containing extruded soybean when compared to control. The organic selenium supplementation increased (P<0,05) milk selenium concentration and delayed milk oxidation measured by peroxide value (P<0,01). The inclusion of 21% of extruded soybean in dairy cattle diet resulted in production of milk fat with modified fatty acid profile with lower concentration of saturated fatty acids and higher concentration of polyunsaturated fatty acids and CLA content. However the milk fat was more susceptible to oxidation. The organic selenium supplementation was efficient to overcome this problem and therefore could maintain the stability of milk.

A formulação de dietas para ruminantes pode alterar o perfil de ácidos do leite, potencializando o valor nutritivo e nutracêutico do mesmo. Estas alterações podem, no entanto, aumentar a susceptibilidade do leite à oxidação, perdendo qualidade em sabor e tempo de prateleira. O objetivo deste trabalho consistiu em avaliar o efeito do fornecimento de dieta com alto teor de soja integral extrusada, associada ou não à suplementação com selênio orgânico, sobre a produção, a composição, a concentração de CLA, o perfil de ácidos graxos e a estabilidade oxidativa do leite. Vinte e quatro vacas holandesas foram agrupadas em três blocos, de acordo com o número de lactações e alimentadas com dieta total contendo 56% de volumoso e 44% de concentrado. Os animais foram distribuídos aleatoriamente nos seguintes tratamentos: A) dieta controle, B) dieta contendo 21% de soja extrusada e C) dieta contendo 21% de soja extrusada + 5 mg de selênio orgânico. As dietas foram balanceadas para obtenção dos mesmos níveis de energia e proteína. O fornecimento de soja extrusada não afetou o consumo de matéria seca, o peso vivo dos animais e a produção de leite. As vacas alimentadas com soja extrusada produziram leite com menor (P<0,01) porcentagem de gordura, menor (P<0,01) concentração de ácidos graxos de cadeias curta e média e maior (P<0,01) concentração de ácidos graxos de cadeia longa, quando comparadas às vacas alimentadas com a dieta controle. Com relação ao teor de saturação, produziram leite com menor (P<0,01) concentração de ácidos graxos saturados e maior (P<0,01) concentração de ácidos graxos poliinsaturados e de ácido linoléico conjugado (CLA). A concentração de colesterol total foi maior (P<0,01) no soro sanguíneo das vacas alimentadas com soja extrusada, quando comparado ao controle. A suplementação com selênio aumentou (P<0,05) significativamente a concentração do mineral no leite e retardou a oxidação do mesmo, avaliada pelo índice de peróxido (P<0,01). O fornecimento da dieta contendo 21% de soja alterou o perfil de ácidos graxos do leite, diminuindo a concentração de ácidos graxos saturados e aumentando a concentração de ácidos graxos poliinsaturados e ácido linoléico conjugado, no entanto, produziu um leite mais susceptível à oxidação. O enriquecimento do leite com selênio conseguiu minimizar este efeito influenciando positivamente a estabilidade do leite. / The manipulation of ruminant diet can modify the fatty acid profiles increasing its nutritive and therapeutic value. However, this change in fatty acid composition can increase milk oxidation and can affect its flavor and shelf life. The objective of this study was to evaluate the effect of extruded soybean diet and organic selenium supplementation on milk composition, CLA content, fatty acid profiles and oxidative stability of milk. Twenty four Holstein cows were grouped into three blocks according to number of parturition and were fed diets containing 56% forage and 44% concentrate. The animals were allocated to three treatments: A) control diet, B) 21% extruded soybean diet and C) 21% extruded soybean diet + 5 mg of organic selenium. The diets were formulated to provide the same energy and protein levels. The extruded soybean did not affect the dry matter intake, body weight and milk production. The cows fed extruded soybean diet produced milk with lower (P<0,01) concentration of fat, lower (P<0,01) concentration of short and medium chain fatty acids and higher (P<0,01) concentration of long chain fatty acids when compared to control group. Regarding to saturation level, those animals produced milk with lower (P<0,01) concentration of saturated fatty acids and higher (P<0,01) concentration of polyunsaturated fatty acids and CLA content. The serum concentration of cholesterol was higher (P<0,01) for treatments containing extruded soybean when compared to control. The organic selenium supplementation increased (P<0,05) milk selenium concentration and delayed milk oxidation measured by peroxide value (P<0,01). The inclusion of 21% of extruded soybean in dairy cattle diet resulted in production of milk fat with modified fatty acid profile with lower concentration of saturated fatty acids and higher concentration of polyunsaturated fatty acids and CLA content. However the milk fat was more susceptible to oxidation. The organic selenium supplementation was efficient to overcome this problem and therefore could maintain the stability of milk.

The theoretical part is concerned with the functional application of selenium and iodine supplementation methods, risks disproportionate to their income, including changes in biochemical and hematological parameters and evaluation methods of adequate saturation. The practical part analyzes the results of experiments on sheep with a defined content of selenium and iodine in the ration evaluation of selected indicators of the metabolic profile of ewes and their lambs. The results are processed statistically. It reflected the dynamics and interactions of monitored parameters.