Neurotransmitter receptor types mediating the generation of intracellular signals are of two types; ligand-gated ion channels and G protein coupled receptors. The effector enzyme phosphoinositide-specific phospholipase C (PLC) is modulated by stimulation of G protein coupled receptors, leading to an increased breakdown of inositol phospholipids ("Ptdlns breakdown").In recent years, the receptors in the brain coupled to PLC and modulation of such receptor-mediated Ptdlns breakdown have been characterised. One such modulation is the "potassium effect", whereby an increase in the assay [K+] from 6 to 18 mM potentiates the Ptdlns breakdown response to the muscarinic agonist carbachol in the rat brain. It has been speculated that this effect is one way of enhancing the signal :noise ratio of muscarinic neurotransmission. The mechanisms responsible for the potassium effect have been studied in this thesis.Initial methodological studies indicated that the temperature of the Krebs buffer used after tissue dissection was an important factor regulating the Ptdlns response to receptor stimulation. Expressing the Ptdlns breakdown response as a fraction of the total labelled phosphoinositides was more useful than other ways of expressing the data. Acid extraction of the Lipid fraction was also superior to neutral extraction.Miniprismspreparedfrompig striatum and hippocampus showed qualitative (but not quantitative) similarities with the rat with respect to stimulation by carbachol, noradrenaline and the potassium effect. Dopamine also stimulated Ptdlns breakdown, though probably via a noradrenergic mechanism.The enhancing actions of potassium appeared to be selective for muscarinic Ml-type receptors. Thus glutamate, quisqualate and NaF-stimulated Ptdlns breakdown are not affected by raised [K+].The potassium effect is brought about by two mechanisms. In calcium-free Krebs buffer, the effect could be mimicked by the calcium channel agonist BAY K-8644 and partially antagonised by verapamil. At an assay [Ca2*] of 2.52 mM, however, modulation of calcium uptake had little effect on carbachol-stimulated Ptdlns breakdown at either normal or raised [K+]. The synergy between potassium and carbachol at252 mM Ca?+ is not dependent upon tissue depolarisation perse, since other ways of depolarising the tissue did not enhance the response to carbachol. It is suggested that potassium might have a direct effect on the muscarinic Ml-type receptor - G protein - PLC complex.In order to investigate this possibility, the effect of fluoride ions (which activate G proteins via formation of AlF4) on basal and carbachol-stimulated Ptdlns breakdown was investigated. Fluoride ions inhibited the enhanced breakdown response to carbachol found at raised [K+]. However, this effect is secondary to effects of fluoride on PLC substrate availibility rather than on G protein function. / digitalisering@umu

Fine tuning a nutritional strategy by incorporating dietary fibre (DF) in pig diets can help to improve gut health. Fermentation of DF, especially the soluble fraction, in pig intestines yields short-chain fatty acids (SCFA) and lactic acid, which have been found to improve gut health by favouring the growth of health-promoting bacteria such as <i>Lactobacilli</i> and <i>Bifidobacteria</i>, at the expense of pathogenic ones like <i>Clostridium</i> or <i>Salmonella</i>, which may enhance the health of host species. The presence of fermentable fibre in the pig diet can also contribute to reducing nitrogen (N) excretion, which can have a positive impact on the environmental footprint, one of the main concerns of a modern commercial pork production.<p> The overall objective of this thesis project was to evaluate the fermentation characteristics of a selection of feedstuffs in the pig intestines and their potential impact on the gut environment and nitrogen excretion. The evaluation was performed by executing two projects using both <i>in vitro</i> and in <i>vivo studies</i>.<p> The first project focused on the fermentation characteristics of hulless barley in comparison to hulled barley and oats and their effects on the gut environment, especially the production of fermentation metabolites. The rate of fibre fermentation in the intestines was first studied by means of an in vitro gas production technique. The results demonstrated that hulless barleys have higher fermentability and produce higher amounts of SCFA than hulled barley and oats. An experiment carried out on pigs confirmed that the fermentation of the soluble fibre fraction of hulless barley in the gut leads to increased production of SCFA and lactic acid, which in turn contribute to the growth of potentially beneficial microbiota and decrease potentially harmful bacteria, an indicator of improved gut health. This finding shows that gut health parameters may be modulated. Thus gut health could potentially be improved through feed formulation by a judicious selection of feed ingredients with specific fibre fractions, not only by the addition of isolated fibres, which is commonly recommended at present.<p> The second project was executed to study the effect of some feedstuffs differing in their DF and protein content on fermentation characteristics and N excretion in pigs. The feedstuffs included wheat bran, wood cellulose, peas, pea hulls, pea inner fibre, sugar beet pulp, flax seed meal and corn distiller's dried grains with solubles. The results showed that peas and pea fibre-based diets produced higher amounts of SCFA and reduced N excreted, compared to others. In a parallel <i>in vitro</i> study, fermentation characteristics and bacterial protein synthesis was also studied using the same feed ingredients. The findings of the <i>in vitro</i> study corraborated the results of the <i>in vivo</i> experiment. These studies showed that peas and pea fibres have the potential to be used in pig diets in order to gain gut health-benefits and reduce N excretion.<p> From this thesis, it can be concluded that sources and type of dietary fibre have a significant effect on the production of fermentation metabolites in the pig intestine and on N excretion. Among the feed ingredients studied, hulless barley and pea fibres seem to have the greatest potential to be included in pig diets as a source of fermentable fibre to modulate the gut environment, which in turn, extend possibly health-promoting properties and reduce N excretion from pigs. However, further research is needed to understand the specific health benefits of these fibre sources and to quantify the specific fibre components required to achieve these benefits.

The nutritional value of flaxseed meal (FSM), a by-product of the flax crushing industry, has not been evaluated properly for use within swine rations. A series of experiments were conducted to determine the nutritional profile of this novel feed ingredient for pigs.<p>The analysis of FSM revealed that it contains, on a dry matter (DM) basis, 133 g/kg ether extract (EE), 345 g/kg crude protein (CP), 60 g/kg ash, 164 g/kg ADF, 250 g/kg NDF, 102 g/kg crude fibre, 14 g/kg starch and 9 g/kg phosphorus. The gross energy (GE) content of the meal was 5.2 Mcal/kg DM. The ether extract fraction was characterized by, as a percent of total fat, 46.6% á-linolenic acid, an omega-3 fatty acid. Palmitic, stearic, oleic and linoleic acids accounted for 9.5, 4.8, 20.7 and 18.4% of the total fat content respectively. The crude protein content was well balanced for all amino acids with the exception of lysine (4.1% of CP), the level of which falls below that of the requirements for growing pigs (5.3% of CP for pigs 20-50 kg). The apparent digestibility of DM, nitrogen, ash, EE and GE as well as determination of the DE and NE content of FSM was determined for both growing pigs (32 pigs, initial weight 70 ± 3 kg) and gestating sows (26 pigs, parities 2 4). Animals were fed wheat/barley based diets containing 0, 10, 20 or 30% FSM. Faecal grab samples were collected for 3 days after a dietary adaptation period. The apparent digestibility of nutrients in FSM was determined both by regression and by difference calculations. As calculated by difference, the apparent digestibility coefficients for DM, nitrogen, ash, and GE were 63.0, 60.8, 22.3 and 60.5% respectively for growing pigs. The values obtained for sows were 64.1, 58.8, 20.8, 94.9 and 65.4% for DM, nitrogen, ash, EE and GE respectively. The DE content was 3.37 Mcal/kg for growing pigs and 3.52 Mcal/kg for sows. Net energy was then estimated by use of a prediction equation to be 2.34 and 2.44 Mcal/kg for growing pigs and sows. <p>An experiment was conducted to evaluate the growth performances and carcass fatty acid profiles of pigs fed with graded levels of FSM. A total of 200 pigs (100 barrows, 100 gilts; initial weight 32 ± 4 kg) were blocked by gender and housed in groups of 5 pigs per pen. The experiment was divided into three phases for pigs 32-60 kg, 60-85 kg and 85-115 kg. Each group was assigned to one of four dietary treatments containing 0, 5, 10 or 15% FSM at the expense of wheat and soybean meal. At the time of market, 6 pigs per treatment group were randomly selected for carcass fatty acid analysis, and backfat and rib-end loin samples were collected. The average daily gains, average daily feed intakes and gain to feed ratios were not affected by dietary treatment (P > 0.05). Inclusion of 15% dietary FSM increased the ALA content from 11 to 47 (± 0.8) mg/g of backfat (P < 0.001) and from 5 to 10 (± 0.4) mg/g of loin tissue (P < 0.001). Increasing dietary FSM decreased the saturated fatty acid content of backfat (P < 0.01). <p> The final experiment was designed to determine the availability of phosphorus in semi-synthetic diets containing FSM, and to determine the effects of microbial phytase inclusion of this availability. Five treatment groups, 8 barrows (45 ± 4 kg initial weight) each, were fed a diet containing 30% FSM with increasing levels of phytase (0, 575, 1185, 2400 and 2570 FTU/kg). Apparent P digestibility increased from 20.6 to 61.3% with the inclusion of up to 2570 FTU/kg microbial phytase (P < 0.001), and followed a quadratic response pattern with an R2 value of 0.96. A broken-line analysis estimated the optimal phytase inclusion level to be 1415 FTU/kg of diet. Inclusion of just 575 FTU/kg accounted for half of the response, improving the apparent P digestibility by 20% and reducing P excretion by 850 mg/kg dry matter intake.

Lung dysfunction due to exposure to air in high intensity livestock barn operations is a common problem for workers in these facilities. Exposure to this air has been linked to disorders such as chronic bronchitis, occupational asthma, organic dust toxic syndrome, and chronic cough and phlegm. These symptoms have been linked to higher levels of endotoxins in air in chicken and swine barns. However, there are many other toxic molecules such as bacterial DNA and gases capable of inducing respiratory inflammation. Bacterial molecules are recognized through highly conserved pattern recognition molecules called Toll-like receptors (TLR). While lipopolysaccharides are recognized by TLR4, bacterial unmethylated DNA binds to and signals through TLR9. As a prelude to understanding the biology of TLR9 in lung inflammation, it is important to precisely clarify their in situ expression in the lung. I determined expression of TLR9 in intact lungs from cattle, pigs, dogs, horses, mice, and humans. Two samples from normal lungs of cattle, pigs, dogs, three from horses, and two from inflamed calf lungs were tested. Five normal mouse and three normal human lungs were similarly tested as well as 5 human lungs with diagnosis of asthma. The expression was determined with multiple methods such as Western blots, immunohistology, immunogold electron microscopy and in situ hybridization. Lungs from all the species showed TLR9 expression in the bronchial epithelium, vascular endothelium, alveolar septa, alveolar macrophages, and type-II alveolar epithelial cells. Immuno-electron microscopy detected TLR9 on the plasma membrane, cytoplasm and the nucleus of various cells including macrophages. In situ hybridization demonstrated TLR9 mRNA in the bronchial epithelium, vascular endothelium, alveolar septa, alveolar macrophages, and type-II alveolar epithelial cells of mouse and human. Asthmatic human lungs showed many more inflammatory cells expressing TLR9 compared to healthy lungs. In cattle and horses, pulmonary intravascular macrophages showed robust expression of TLR9. Depletion of pulmonary intravascular macrophages in horses resulted in significant reduction in total TLR9 mRNA in the lungs. Having determined that TLR9 expression is similarly expressed on many lung cell types in mice and humans, I determined the role of TLR9 in barn air induced lung inflammation by exposing TLR9-/- and wild-type mice (6 per group) to single or multiple days (5 and 20) in a chicken barn. Each exposure was of 8 hours/day duration. The TLR9-/- mice exposed 5 and 20 times showed significant reductions in TNF-alpha and IFN-gamma expression in lung lavages as well as cellular changes consistent with reduced lung inflammation such as reductions in the number of lung neutrophils. This suggests that barn dust DNA, acting through TLR9, contributes to lung inflammation seen in response to exposure to chicken barn air. These fundamental data advance our knowledge on the cell-specific expression of TLR9 across a range of species including the humans and demonstrate that TLR9-/- partially regulates lung inflammation induced following exposure to chicken barn air.

Lung dysfunction due to exposure to air in high intensity livestock barn operations is a common problem for workers in these facilities. Exposure to this air has been linked to disorders such as chronic bronchitis, occupational asthma, organic dust toxic syndrome, and chronic cough and phlegm. These symptoms have been linked to higher levels of endotoxins in air in chicken and swine barns. However, there are many other toxic molecules such as bacterial DNA and gases capable of inducing respiratory inflammation. Bacterial molecules are recognized through highly conserved pattern recognition molecules called Toll-like receptors (TLR). While lipopolysaccharides are recognized by TLR4, bacterial unmethylated DNA binds to and signals through TLR9. As a prelude to understanding the biology of TLR9 in lung inflammation, it is important to precisely clarify their in situ expression in the lung. I determined expression of TLR9 in intact lungs from cattle, pigs, dogs, horses, mice, and humans. Two samples from normal lungs of cattle, pigs, dogs, three from horses, and two from inflamed calf lungs were tested. Five normal mouse and three normal human lungs were similarly tested as well as 5 human lungs with diagnosis of asthma. The expression was determined with multiple methods such as Western blots, immunohistology, immunogold electron microscopy and in situ hybridization. Lungs from all the species showed TLR9 expression in the bronchial epithelium, vascular endothelium, alveolar septa, alveolar macrophages, and type-II alveolar epithelial cells. Immuno-electron microscopy detected TLR9 on the plasma membrane, cytoplasm and the nucleus of various cells including macrophages. In situ hybridization demonstrated TLR9 mRNA in the bronchial epithelium, vascular endothelium, alveolar septa, alveolar macrophages, and type-II alveolar epithelial cells of mouse and human. Asthmatic human lungs showed many more inflammatory cells expressing TLR9 compared to healthy lungs. In cattle and horses, pulmonary intravascular macrophages showed robust expression of TLR9. Depletion of pulmonary intravascular macrophages in horses resulted in significant reduction in total TLR9 mRNA in the lungs. Having determined that TLR9 expression is similarly expressed on many lung cell types in mice and humans, I determined the role of TLR9 in barn air induced lung inflammation by exposing TLR9-/- and wild-type mice (6 per group) to single or multiple days (5 and 20) in a chicken barn. Each exposure was of 8 hours/day duration. The TLR9-/- mice exposed 5 and 20 times showed significant reductions in TNF-alpha and IFN-gamma expression in lung lavages as well as cellular changes consistent with reduced lung inflammation such as reductions in the number of lung neutrophils. This suggests that barn dust DNA, acting through TLR9, contributes to lung inflammation seen in response to exposure to chicken barn air. These fundamental data advance our knowledge on the cell-specific expression of TLR9 across a range of species including the humans and demonstrate that TLR9-/- partially regulates lung inflammation induced following exposure to chicken barn air.

To investigate whether piglets show different personality types due to teat position, a number ofexperiments were performed on 21 different litters, 9 to 31 days old. The tests included threefocal piglets in each litter with different teat position, anterior, middle and posterior, inobservations on suckling behaviour, undisturbed behaviours, a novel object test and a straw-testwhere thirteen behaviours were observed. The tests found an effect on teat position on frequencyof disputes between piglets during suckling and their teat position (P=0.018) and a tendency ofdifference in undisturbed playing behaviour between piglets at different teat position (P=0.057).All tests found differences between litters concerning behaviours. With a Principal componentanalysis, the four personality types curiosity, playful, food interest and straw interest wasestablished. No effect between different teat positions and personality type were found, but atendency of difference between personality trait playful and different teat positions (P=0.062). Inconclusion: no significant differences between teat position and personality type in piglets couldbe established.

The objective of this study was to determine whether piglets from different positions in the teat order differ inpersonality. Teat order is established within the first days after birth and from that time, piglets basically suckle fromthe same teat pair until weaning. Three piglets from each of 21 litters (n = 63) were chosen for their place in teatorder; one from anterior teats (A), one from middle teats (B), and one from rear teats (C). The piglets were observedduring suckling, undisturbed behaviour, a novel object test and a straw test. A total of 13 behaviours were registered.A significant difference (P &lt; 0.05) between A, B and C piglets was found in the frequency of disputes duringsuckling, where B piglets were most involved in disputes and C piglets least involved, and a tendency (P &lt; 0.1) wasfound in frequency of undisturbed playing and exploration. A piglets played and explored most, and C piglets least.Four interesting components were found with principal component analysis, explaining 62.4 % of the total variation.These represent personality traits tentatively named exploration, playfulness, food interest and straw interest. Therewere no significant differences in personality type between A, B and C piglets although a tendency that A pigletswere most playful and C piglets were least playful was found. Hence, this study suggests that there might be acorrelation between teat order position and personality.

The nutritional value of flaxseed meal (FSM), a by-product of the flax crushing industry, has not been evaluated properly for use within swine rations. A series of experiments were conducted to determine the nutritional profile of this novel feed ingredient for pigs.<p>The analysis of FSM revealed that it contains, on a dry matter (DM) basis, 133 g/kg ether extract (EE), 345 g/kg crude protein (CP), 60 g/kg ash, 164 g/kg ADF, 250 g/kg NDF, 102 g/kg crude fibre, 14 g/kg starch and 9 g/kg phosphorus. The gross energy (GE) content of the meal was 5.2 Mcal/kg DM. The ether extract fraction was characterized by, as a percent of total fat, 46.6% á-linolenic acid, an omega-3 fatty acid. Palmitic, stearic, oleic and linoleic acids accounted for 9.5, 4.8, 20.7 and 18.4% of the total fat content respectively. The crude protein content was well balanced for all amino acids with the exception of lysine (4.1% of CP), the level of which falls below that of the requirements for growing pigs (5.3% of CP for pigs 20-50 kg). The apparent digestibility of DM, nitrogen, ash, EE and GE as well as determination of the DE and NE content of FSM was determined for both growing pigs (32 pigs, initial weight 70 ± 3 kg) and gestating sows (26 pigs, parities 2 4). Animals were fed wheat/barley based diets containing 0, 10, 20 or 30% FSM. Faecal grab samples were collected for 3 days after a dietary adaptation period. The apparent digestibility of nutrients in FSM was determined both by regression and by difference calculations. As calculated by difference, the apparent digestibility coefficients for DM, nitrogen, ash, and GE were 63.0, 60.8, 22.3 and 60.5% respectively for growing pigs. The values obtained for sows were 64.1, 58.8, 20.8, 94.9 and 65.4% for DM, nitrogen, ash, EE and GE respectively. The DE content was 3.37 Mcal/kg for growing pigs and 3.52 Mcal/kg for sows. Net energy was then estimated by use of a prediction equation to be 2.34 and 2.44 Mcal/kg for growing pigs and sows. <p>An experiment was conducted to evaluate the growth performances and carcass fatty acid profiles of pigs fed with graded levels of FSM. A total of 200 pigs (100 barrows, 100 gilts; initial weight 32 ± 4 kg) were blocked by gender and housed in groups of 5 pigs per pen. The experiment was divided into three phases for pigs 32-60 kg, 60-85 kg and 85-115 kg. Each group was assigned to one of four dietary treatments containing 0, 5, 10 or 15% FSM at the expense of wheat and soybean meal. At the time of market, 6 pigs per treatment group were randomly selected for carcass fatty acid analysis, and backfat and rib-end loin samples were collected. The average daily gains, average daily feed intakes and gain to feed ratios were not affected by dietary treatment (P > 0.05). Inclusion of 15% dietary FSM increased the ALA content from 11 to 47 (± 0.8) mg/g of backfat (P < 0.001) and from 5 to 10 (± 0.4) mg/g of loin tissue (P < 0.001). Increasing dietary FSM decreased the saturated fatty acid content of backfat (P < 0.01). <p> The final experiment was designed to determine the availability of phosphorus in semi-synthetic diets containing FSM, and to determine the effects of microbial phytase inclusion of this availability. Five treatment groups, 8 barrows (45 ± 4 kg initial weight) each, were fed a diet containing 30% FSM with increasing levels of phytase (0, 575, 1185, 2400 and 2570 FTU/kg). Apparent P digestibility increased from 20.6 to 61.3% with the inclusion of up to 2570 FTU/kg microbial phytase (P < 0.001), and followed a quadratic response pattern with an R2 value of 0.96. A broken-line analysis estimated the optimal phytase inclusion level to be 1415 FTU/kg of diet. Inclusion of just 575 FTU/kg accounted for half of the response, improving the apparent P digestibility by 20% and reducing P excretion by 850 mg/kg dry matter intake.

Fine tuning a nutritional strategy by incorporating dietary fibre (DF) in pig diets can help to improve gut health. Fermentation of DF, especially the soluble fraction, in pig intestines yields short-chain fatty acids (SCFA) and lactic acid, which have been found to improve gut health by favouring the growth of health-promoting bacteria such as <i>Lactobacilli</i> and <i>Bifidobacteria</i>, at the expense of pathogenic ones like <i>Clostridium</i> or <i>Salmonella</i>, which may enhance the health of host species. The presence of fermentable fibre in the pig diet can also contribute to reducing nitrogen (N) excretion, which can have a positive impact on the environmental footprint, one of the main concerns of a modern commercial pork production.<p> The overall objective of this thesis project was to evaluate the fermentation characteristics of a selection of feedstuffs in the pig intestines and their potential impact on the gut environment and nitrogen excretion. The evaluation was performed by executing two projects using both <i>in vitro</i> and in <i>vivo studies</i>.<p> The first project focused on the fermentation characteristics of hulless barley in comparison to hulled barley and oats and their effects on the gut environment, especially the production of fermentation metabolites. The rate of fibre fermentation in the intestines was first studied by means of an in vitro gas production technique. The results demonstrated that hulless barleys have higher fermentability and produce higher amounts of SCFA than hulled barley and oats. An experiment carried out on pigs confirmed that the fermentation of the soluble fibre fraction of hulless barley in the gut leads to increased production of SCFA and lactic acid, which in turn contribute to the growth of potentially beneficial microbiota and decrease potentially harmful bacteria, an indicator of improved gut health. This finding shows that gut health parameters may be modulated. Thus gut health could potentially be improved through feed formulation by a judicious selection of feed ingredients with specific fibre fractions, not only by the addition of isolated fibres, which is commonly recommended at present.<p> The second project was executed to study the effect of some feedstuffs differing in their DF and protein content on fermentation characteristics and N excretion in pigs. The feedstuffs included wheat bran, wood cellulose, peas, pea hulls, pea inner fibre, sugar beet pulp, flax seed meal and corn distiller's dried grains with solubles. The results showed that peas and pea fibre-based diets produced higher amounts of SCFA and reduced N excreted, compared to others. In a parallel <i>in vitro</i> study, fermentation characteristics and bacterial protein synthesis was also studied using the same feed ingredients. The findings of the <i>in vitro</i> study corraborated the results of the <i>in vivo</i> experiment. These studies showed that peas and pea fibres have the potential to be used in pig diets in order to gain gut health-benefits and reduce N excretion.<p> From this thesis, it can be concluded that sources and type of dietary fibre have a significant effect on the production of fermentation metabolites in the pig intestine and on N excretion. Among the feed ingredients studied, hulless barley and pea fibres seem to have the greatest potential to be included in pig diets as a source of fermentable fibre to modulate the gut environment, which in turn, extend possibly health-promoting properties and reduce N excretion from pigs. However, further research is needed to understand the specific health benefits of these fibre sources and to quantify the specific fibre components required to achieve these benefits.

In sächsischen Schweinehaltungsbetrieben wurde ein objektives Bewertungssystem zur Tierhygiene und Tiergesundheit entwickelt und erprobt. Aus zahlreichen Untersuchungen in den Produktionseinheiten Ferkelerzeugung, Ferkelaufzucht, Jungsauenaufzucht und Mast resultieren Kennziffern zur Hygiene, Klinik und Gesundheit, die einen Vergleich verschiedener Bestände erlauben. Sie lassen stufenübergreifend auch eine Bewertung einzelner Produktionseinheiten zu. Mit regelmäßigen Untersuchungen ist zudem der zeitliche Verlauf der Kennziffern und damit des Hygiene- und Gesundheitsstatus möglich.