Process Analytical Technology Approach on Fluid Bed Granulation and Drying : Identifying Critical Relationships and Constructing the Design Space

Fluid bed granulation is a key pharmaceutical process which improves many of the powder properties for tablet compression. Dry mixing, wetting and drying phases are included in the fluid bed granulation process. Granules of high quality can be obtained by understanding and controlling the critical process parameters by timely measurements. Physical process measurements and particle size data of a fluid bed granulator that are analysed in an integrated manner are included in process analytical technologies (PAT). Recent regulatory guidelines strongly encourage the pharmaceutical industry to apply scientific and risk management approaches to the development of a product and its manufacturing process. The aim of this study was to utilise PAT tools to increase the process understanding of fluid bed granulation and drying. Inlet air humidity levels and granulation liquid feed affect powder moisture during fluid bed granulation. Moisture influences on many process, granule and tablet qualities. The approach in this thesis was to identify sources of variation that are mainly related to moisture. The aim was to determine correlations and relationships, and utilise the PAT and design space concepts for the fluid bed granulation and drying. Monitoring the material behaviour in a fluidised bed has traditionally relied on the observational ability and experience of an operator. There has been a lack of good criteria for characterising material behaviour during spraying and drying phases, even though the entire performance of a process and end product quality are dependent on it. The granules were produced in an instrumented bench-scale Glatt WSG5 fluid bed granulator. The effect of inlet air humidity and granulation liquid feed on the temperature measurements at different locations of a fluid bed granulator system were determined. This revealed dynamic changes in the measurements and enabled finding the most optimal sites for process control. The moisture originating from the granulation liquid and inlet air affected the temperature of the mass and pressure difference over granules. Moreover, the effects of inlet air humidity and granulation liquid feed rate on granule size were evaluated and compensatory techniques used to optimize particle size. Various end-point indication techniques of drying were compared. The ∆T method, which is based on thermodynamic principles, eliminated the effects of humidity variations and resulted in the most precise estimation of the drying end-point. The influence of fluidisation behaviour on drying end-point detection was determined. The feasibility of the ∆T method and thus the similarities of end-point moisture contents were found to be dependent on the variation in fluidisation between manufacturing batches. A novel parameter that describes behaviour of material in a fluid bed was developed. Flow rate of the process air and turbine fan speed were used to calculate this parameter and it was compared to the fluidisation behaviour and the particle size results. The design space process trajectories for smooth fluidisation based on the fluidisation parameters were determined. With this design space it is possible to avoid excessive fluidisation and improper fluidisation and bed collapse. Furthermore, various process phenomena and failure modes were observed with the in-line particle size analyser. Both rapid increase and a decrease in granule size could be monitored in a timely manner. The fluidisation parameter and the pressure difference over filters were also discovered to express particle size when the granules had been formed. The various physical parameters evaluated in this thesis give valuable information of fluid bed process performance and increase the process understanding.

Vikuroivia vilkaisuja : Ruumis, sukupuoli, seksuaalisuus ja visuaalisen kulttuurin tutkimus

The PhD dissertation "Bucking Glances: On Body, Gender, Sexuality and Visual Culture Research" consists of theoretical introduction and five articles published between 2002-2005. The articles analyze the position of visual representations in the processes of knowledge production on acceptable genders, bodies, and sexualities in contemporary Wes¬tern societies. The research material is heterogeneous, consisting of representations of contemporary art, advertisements, and fashion images. The ideological starting point of the PhD dissertation is the politics of the gaze and the methods used to expose this are the concepts of oppositional gaze, close reading, and resisting reading. The study situates visual representations in dialogue with the concepts of the grotesque and androgyny, as well as with queer-theory and theories of the gaze. The research challenges normative meanings of visual representations and opens up space for more non-conventional readings attached to femininity and masculinity. The visual material is read as troubling the prevailing heteronormative gender system. The dissertation also indicates how visual culture research utilizing the approach of queer theory can be fruitful in opposing and re-visioning changes in the repressive gender system. The article "A Heroic Male and A Beautiful Woman. Teemu Mäki, Orlan and the Ambivalence of the Grotesque Body" problematizes the concept of heroic masculinity through the analysis of the Finnish artist Teemu Mäki's masochistic performance The Good Friday (1989). It also analyzes cosmetic surgery, undertaken by the French artist Orlan, as a cultural tool in constructing and visualizing the contemporary, com¬mercial ideals of female beauty. The article "Boys Will Be Girls Will Be Boys Will Be Girls. The Ambivalence of Androgyny in Calvin Klein' Advertisements" is a close reading of the Calvin Klein perfume advertisement One (1998) in reference to the concept of androgyny. The critical point of the article is that androgynous male bodies allow the extension of the categorical boundaries of masculinity and homosexuality, whereas representations of androgynous women feed into the prevailing stereotypes of femininity, namely the fear of fat. The article "See-through Closet: Female Androgyny in the 1990s Fashion Images, New Woman and Lesbian Chic" analyzes the late 1990s fashion advertisements through the concept of female androgyny. The article argues that the figures of the masculine female androgynes in the late 1990s fashion magazines do not problematize the dichotomous gender binary. The women do not pass as men but produce a variation of heterosexual desirability. At the same time, the representations open up space for lesbian gazing and desiring. The article "Why are there no lesbian advertisements?" addresses the issue of femme gaze and desire in relation to heterosexual fashion advertisements from the British edition of the mainstream fashion magazine Vogue. The article considers possibilities for resistant femme visibility, identification, and desire. The article "Woman, Food, Home. Pirjetta Brander's and Heidi Romo's Works as Bucking Representations of Femininity" analyses the production and queering of heteronormative femininity and family through the analysis of art works. The article discusses how the term queer has been translated into Finnish. The article also introduces a new translation for the term queer: the noun vikuuri, i.e. faulty form and the verb vikuroida, i.e. to buck. In Finnish, the term vikuuri is the vernacular or broken form of the term figure, i.e. figuuri. Vikuuri represents all forms situated outside the norm and the normative.

Molecular Background of Common Dyslipidemias

The leading cause of death in the Western world continues to be coronary heart disease (CHD). At the root of the disease process is dyslipidemia an aberration in the relevant amounts of circulating blood lipids. Cholesterol builds up in the arterial wall and following rupture of these plaques, myocardial infarction or stroke can occur. Heart disease runs in families and a number of hereditary forms are known. The leading cause of adult dyslipidemia presently however is overweight and obesity. This thesis work presents an investigation of the molecular genetics of common, hereditary dyslipidemia and the tightly related condition of obesity. Familial combined hyperlipidemia (FCHL) is the most common hereditary dyslipidemia in man with an estimated population prevalence of 1-6%. This complex disease is characterized by elevated levels of serum total cholesterol, triglycerides or both and is observed in about 20% of individuals with premature CHD. Our group identified the disease to be associated with genetic variation in the USF1 transcription factor gene. USF1 has a key role in regulating other genes that control lipid and glucose metabolism as well as the inflammatory response all central processes in the progression of atherosclerosis and CHD. The first two works of this thesis aimed at understanding how these USF1 variants result in increased disease risk. Among the many, non-coding single-nucleotide polymorphisms (SNPs) that associated with the disease, one was found to have a functional effect. The risk-enhancing allele of this SNP seems to eradicate the ability of the important hormone insulin to induce the expression of USF1 in peripheral tissues. The resultant changes in the expression of numerous USF1 target genes over time probably enhance and accelerate the atherogenic processes. Dyslipidemias often represent an outcome of obesity and in the final work of this thesis we wanted to address the metabolic pathways related to acquired obesity. It is recognized that active processes in adipose tissue play an important role in the development of dyslipidemia, insulin resistance and other pathological conditions associated with obesity. To minimize the confounding effects of genetic differences present in most human studies, we investigated a rare collection of identical twins that differed significantly in the amount of body fat. In the obese, but otherwise healthy young adults, several notable changes were observed. In addition to chronic inflammation, the adipose tissue of the obese co-twins was characterized by a marked (47%) decrease in amount of mitochondrial DNA (mtDNA) a change associated with mitochondrial dysfunction. The catabolism of branched chain amino acids (BCAAs) was identified as the most down-regulated process in the obese co-twins. A concordant increase in the serum level of these insulin secretagogues was identified. This hyperaminoacidemia may provide the feed-back signal from insulin resistant adipose tissue to the pancreas to ensure an appropriately augmented secretory response. The down regulation of BCAA catabolism correlated closely with liver fat accumulation and insulin. The single most up-regulated gene (5.9 fold) in the obese co-twins was osteopontin (SPP1) a cytokine involved in macrophage recruitment to adipose tissue. SPP1 is here implicated as an important player in the development of insulin resistance. These studies of exceptional study samples provide better understanding of the underlying pathology in common dyslipidemias and other obesity associated diseases important for future improvement of intervention strategies and treatments to combat atherosclerosis and coronary heart disease.

Concentrate feeding strategies for growing and finishing dairy bulls offered grass silage-based diets

The first aim of this thesis was to produce data for evaluating, developing and recommending biologically and economically efficient energy and protein feeding strategies for growing and finishing dairy bulls offered grass silage-based diets. The second aim was to calculate the energy and protein supplies of the dairy bulls fed different grass silage-cereal-based diets and, based on this, to estimate the possible need to revise the current Finnish energy and protein recommendations for growing dairy bulls. The third aim was to demonstrate the phosphorus supply of dairy bulls fed grass silage-cereal-based diets with or without protein supplementation in relation to current feeding recommendations for phosphorus. The results indicate that protein supplement is not needed for finishing dairy bulls (live weight more than 250 kg) when they are fed good-quality grass silage (digestible organic matter more than 650 g/kg dry matter, restricted fermentation with low concentrations of fermentation acids and ammonia N) and grain-based concentrate with a moderate (300-700 g/kg dry matter) concentrate level. The results also suggest that with total mixed ration feeding it is possible to use rather high concentrate proportions (700 g/kg dry matter) in feeding dairy bulls. According to this study, barley fibre is a suitable energy supplement with good-quality silage for growing dairy bulls. The results suggest that 50% of barley grain can be replaced with barley fibre without affecting growth. Also oats is a suitable energy supplement for dairy bulls. However, as a consequence of decreased energy intake, the gain and feed conversion of the bulls were slightly reduced in this study when barley grain was replaced by oats in the diet. Ultimately, the rationality of the use of barley fibre and oats in the future will depend on the price in relation to other concentrates. During the feeding experiments the calculated supply of energy was 10% higher than in the Finnish feeding recommendations for the present growth rate. This indicates that there is a need to update the Finnish feeding recommendations for dairy-breed growing bulls, and further calculations are needed for the energy supply of growing dairy bulls. The calculated supply of AAT (amino acids absorbed from the small intestine) was 38% higher than in the Finnish feeding recommendations for the present growth. Possibly, the present AAT-PBV system is not an optimal protein evaluation system for growing dairy bulls more than 250 kg live weight. The calculations based on the feeding experiments and the Finnish feeding recommendations indicate that in most cases the dairy bulls (live weight more than 250 kg) received enough P from the basic grass silage cereal-based diets without additional mineral feeds. Therefore there is no need to add P in the form of mineral mixtures.

Egg production in furnished cages

In the European Union, conventional cages for laying hens will be faded out at the beginning of 2012. The rationale behind this is a public concern over animal welfare in egg production. As alternatives to conventional cages, the European Union Council Directive 1999/74/EC allows non-cage systems and enriched (furnished) cages. Layer performance, behavior, and welfare in differently sized furnished cages have been investigated quite widely during recent decades, but nutrition of hens in this production system has received less attention. This thesis aims to compare production and feed intake of laying hens in furnished and conventional cages and to study the effects of different dietary treatments in these production systems, thus contributing to the general knowledge of furnished cages as an egg production system. A furnished cage model for 8 hens was compared with a 3-hen conventional cage. Three consecutive experiments each studied one aspect of layer diet: The first experiment investigated the effects of dietary protein/energy ratio, the second dietary energy levels, and the third the effects of extra limestone supplementation. In addition, a fourth experiment evaluated the effects of perches on feed consumption and behavior of hens in furnished cages. The dietary treatments in experiments 1 3 generally had similar effects in the two cage types. Thus, there was no evidence supporting a change in nutrient requirements for laying hens when conventional cages are replaced with small-group furnished cages. Moreover, the results from nutritional experiments conducted in conventional cages can be applied to small-group furnished cage systems. These results support the view that production performance comparable with conventional cages can be achieved in furnished cages. All of the advantages of cages for bird welfare are sustained in the small-group furnished cages used here. In addition, frequent use of perches and nests implies a wider behavioral repertoire in furnished cages than in conventional cages. The increase observed in bone ash content may improve bird welfare in furnished cages. The presence of perches diminished feed consumption during the prelaying period and enhanced the feed conversion ratio during the early laying period in furnished cages. However, as the presence or absence of perches in furnished cages had no significant effect on feed consumption after the prelaying period, the lower feed consumption observed in furnished cages than in conventional cages could be attributed to other factors, such as the presence of wood shavings or a nest box. The wider feed trough space per hen in conventional than in furnished cages may partly explain the higher feed consumption observed in conventional cages.

Northern challanges for profitable production of quality naked oat

Naked oat (Avena sativa f.sp. nuda L.) is the highest quality cereal in northern growing conditions. However the cultivation area of naked oat is remarkably small. Major challenges for naked oat production are to observe its nakedness. The caryopsis of naked oat is sensitive to mechanical damage at harvest, especially at high grain moisture content. The greater the grain moisture content of naked oat at harvest, the more loses of germination capacity was caused by threshing. For producing high quality naked oat seed, it is recommended that harvesting be done at as low grain moisture content as possible. However, if this is not possible, better germination can be ensure with gentle harvest by reducing the cylinder speed. In spite of conventional oat s excellent fat and amino acid composition in animal feed use, as far as nutritional value is concerned, the total energy yield of oat is weaker than other cereals because of the hulls. Also with naked oat the dehulling is not complete, while hull content on different cultivars mostly varied between one to six percent. In addition to genotype, environmental conditions markedly control the expression of nakedness. Thresher settings had only limited effects on hull content. The function of hulls is to protect the groat, but this was confirmed only for Finnish, small grain, cultivar Lisbeth. The oat kernel is generally covered with fine silky hairs termed trichomes. The trichomes of naked oat are partly lost during threshing and handling of grains. Trichomes can cause itchiness in those handling the grains and also accumulate and form fine dust and can block-up machinery. The cultivars differed considerably in pubescence. Some thresher settings, including increased cylinder speed, slightly increased grain polishing such that grains had some areas completely free of trichomes. Adjusting thresher settings was generally not an efficient means of solving the problems associated with naked oat trichomes. The main differences in cultivation costs between naked and conventional oat lie in the amount of seeds required and the drying costs. The main differences affecting the economic result lie in market prices, yield level and feed value. The results indicate that naked oat is financially more profitable than conventional oat, when the crop is sold at a specific price at all yield levels and when the crop is used as feed at highest yield level. At lower yield levels, conventional oat is, in spite of its lower feed value, the more profitable option for feed use. Dehulled oat did not achieve the same economic result as naked oat, as the cost of dehulling, including the hull waste, was considerable. According to this study naked oat can be cultivated successfully under northern conditions, when taking into consideration the soft, naked grain through cultivation chain.

Influence of harvesting strategy on nutrient supply and production of dairy cows consuming diets based on grass and red clover silage

The main objective of this thesis was to elucidate the effects of regrowth grass silage and red clover silage on nutrient supply and milk production of dairy cows as compared with primary growth grass silages. In the first experiment (publication I), two primary growth and four regrowth grass silages were harvested at two stages of growth. These six silages were fed to 24 lactating dairy cows with two levels of concentrate allowance. Silage intake and energy corrected milk yield (ECM) responses, and the range in these response variables between the diets, were smaller when regrowth silages rather than primary growth silages were fed. Milk production of dairy cows reflected the intake of metabolizable energy (ME), and no differences in the ME utilization were found between the diets based on silages harvested from primary growth and regrowth. The ECM response to increased concentrate allowance was, on average, greater when regrowth rather than primary growth silages were fed. In the second experiment (publication II), two silages from primary growth and two from regrowth used in I were fed to rumen cannulated lactating dairy cows. Cows consumed less feed dry matter (DM), energy and protein, and produced less milk, when fed diets based on regrowth silages rather than primary growth silages. Lower milk production responses of regrowth grass silage diets were mainly due to the lower silage DM intake, and could not be accounted for by differences in energy or protein utilization. Regrowth grass silage intake was not limited due to neutral detergent fibre (NDF) digestion or rumen fill or passage kinetics. However, lower intake may be at least partly attributable to plant diseases such as leaf spot infections, dead deteriorating material or abundance of weeds, which are all higher in regrowth compared with primary growth, and increase with advancing regrowth. In the third experiment (publications III and IV), red clover silages and grass silages harvested at two stages of growth, and a mixed diet of red clover and grass silages, were fed to five rumen cannulated lactating dairy cows. In spite of the lower average ME intake for red clover diets, the ECM production remained unchanged suggesting more efficient utilisation of ME for red clover diets compared with grass diets. Intake of N, and omasal canal flows of total non-ammonia N (NAN), microbial and non-microbial NAN were higher for red clover than for grass silage diets, but were not affected by forage maturity. Delaying the harvest tended to decrease DM intake of grass silage and increase that of red clover silage. The digestion rate of potentially digestible NDF was faster for red clover diets than for grass silage diets. Delaying the harvest decreased the digestion rate for grass but increased it for red clover silage diets. The low intake of early-cut red clover silage could not be explained by silage digestibility, fermentation quality, or rumen fill but was most likely related to the nutritionally suboptimal diet composition because inclusion of moderate quality grass silage in mixed diet increased silage DM intake. Despite the higher total amino acid supply of cows fed red clover versus grass silage diets, further milk production responses on red clover diets were possibly compromised by an inadequate supply of methionine as evidenced by lower methionine concentration in the amino acid profile of omasal digesta and plasma. Increasing the maturity of ensiled red clover does not seem to affect silage DM intake as consistently as that of grasses. The efficiency of N utilization for milk protein synthesis was lower for red clover diets than for grass diets. It was negatively related to diet crude protein concentration similarly to grass silage diets.

Rearing strategies of young dairy calves in relation to production, behaviour and welfare

Vasikoiden kasvatus yksilökarsinoissa, imemismahdollisuuden puute maitojuoton yhteydessä sekä pienet juomamäärät ovat tekijöitä, jotka mahdollisesti voivat vähentää vasikoiden hyvinvointia. Vasikoiden kasvatukseen etsitäänkin uusia tapoja, joissa eläinten käyttäytymistarpeet ja hyvinvointi otetaan entistä paremmin huomioon. Tässä väitöskirjassa vasikoiden kasvatusta on tarkasteltu sekä tuotannon että vasikoiden käyttäytymisen ja hyvinvoinnin kannalta. Väitöskirja koostuu kolmesta kokeesta, joista ensimmäisessä tutkittiin vasikoiden kasvatusta ryhmäkarsinoissa ulkona tai sisällä ja vasikoiden kasvatusta sisällä ryhmä- tai yksilökarsinoissa. Toisessa kokeessa vasikoiden annettiin imeä emiään rajoitetusti lypsyn jälkeen viiden tai kahdeksan viikon ajan ja kolmannessa selvitettiin vasikoiden veden juontia, kun vasikat saivat juomarehua vapaasti. Lisäksi kokeiden yhdistetystä aineistosta analysoitiin eri rehujen syöntimäärien suhdetta sekä rehujen vaikutusta kasvuun ennen ja jälkeen maidosta vieroituksen. Tutkimuksessa todettiin, että vasikoita voi ryhmässä kasvattaa kylmissä ja vaihtelevissa sääoloissa ulkona, kunhan ne hoidetaan ja ruokitaan erittäin huolellisesti. Kylmällä ilmalla vasikat saattavat kuitenkin syödä väkirehua vähemmän varsinkin, jos ruokailupaikka on ulkona ja makuualue sisällä. Ryhmässä kasvaneet vasikat aloittivat sekä kuivien rehujen syönnin että märehtimisen nuorempina kuin yksilökarsinassa kasvaneet. Ryhmissä esiintyvää käyttäytymisongelmaa, toisten vasikoiden imemistä, voidaan vähentää hoito- ja ruokintamenetelmillä. Annettaessa vasikoiden imeä emiään rajoitetusti lypsyn jälkeen vasikat oppivat imemään emiään hyvin nopeasti. Lypsytyö vaikeutui muutamien lehmien kohdalla, sillä ne pidättivät maitoa lypsettäessä. Saadessaan imeä rajoitetusti vasikat imivät suurehkoja maitomääriä kerrallaan. Vieroittaminen suurilta maitomääriltä viiden viikon iässä oli kuitenkin liian aikaista, koska vasikat eivät vielä syöneet riittävästi kuivia rehuja. Vieroitus emästä niin viiden kuin kahdeksankin viikon iässä aiheutti vasikoissa levottomuutta ja ääntelyn lisääntymistä. Saadessaan hapatettua juomarehua vapaasti vasikat joivat keskimäärin vain vähän vettä, olipa vesilähteenä avoin ämpäri tai vesinippa. Vasikoiden välillä oli suurta vaihtelua veden juontimäärissä. Viikkoa ennen maidosta vieroitusta vasikat joivat 0-3 l vettä päivässä. Vasikat joivat nipasta kerrallaan vähemmän vettä kuin ämpäristä, ja käyttivät enemmän aikaa päivässä veden juomiseen kuin vesiämpäristä juoneet vasikat. Suurin osa vasikoista joi vettä juomanipoista erikoisella tavalla esimerkiksi painamalla nippaa otsalla ja juomalla tippuvaa vettä. Vesinipat voivat olla siis vasikoille joko vaikeita tai epämukavia käyttää. Vasikoiden juoman maitomäärän lisääntyessä kasvu lisääntyy selvästi. Runsas maidon juominen vähentää kuitenkin kuivien rehujen syöntiä ja vieroitusvaiheessa kasvu voi hidastua. Vasikat olisikin tärkeää vieroittaa vähitellen, ettei muutoksia kasvuun tulisi. Syönti- ja kasvutulokset eivät aina anna oikeaa kuvaa kasvatusmenetelmien eroista eläinten hyvinvoinnin kannalta. Käyttäytyminen on herkkä hyvinvoinnin mittari ja se tulisikin aina huomioida eri kasvatusmenetelmiä arvioitaessa. .

Winter feeding strategies for suckler cows in cold climatic conditions

In Finland, suckler cow production is carried out in circumstances characterized by a long winter period and a short grazing period. The traditional winter housing system for suckler cows has been insulated or uninsulated buildings, but there is a demand for developing less expensive housing systems. In addition, more information is needed on new winter feeding strategies, carried out in inexpensive winter facilities with conventional (hay, grass silage, straw) or alternative (treated straw, industrial by-product, whole-crop silage) feeds. The new feeding techniques should not have any detrimental effects on animal welfare in order to be acceptable to both farmers and consumers. Furthermore, no official feeding recommendations for suckler cows are available in Finland and, thus, recommendations for dairy cows have been used. However, this may lead to over- or underfeeding of suckler cows and, finally, to decreased economic output. In Experiment I, second-calf beef-dairy suckler cows were used to compare the effects of diets based on hay (H) or urea-treated straw (US) at two feeding levels (Moderate; M vs. Low; L) on the performance of cows and calves. Live weight (LW) gain during the indoor feeding was lower for cows on level L than on level M. Cows on diet US lost more LW indoors than those on diet H. The cows replenished the LW losses on good pasture. Calf LW gain and cow milk production were unaffected by the treatments. Conception rate was unaffected by the treatments but was only 69%. Urea-treated straw proved to be a suitable winter feed for spring-calving suckler cows. Experiment II studied the effects of feeding accuracy on the performance of first- and second-calf beef-dairy cows and calves. In II-1, the day-to-day variation in the roughage offered ranged up to ± 40%. In II-2, the same variation was used in two-week periods. Variation of the roughages offered had minor effects on cow performance. Reproduction was unaffected by the feeding accuracy. Accurate feeding is not necessary for young beef-dairy crosses, if the total amount of energy offered over a period of a few weeks fulfills the energy requirements. Effects of feeding strategies with alternative feeds on the performance of mature beef-dairy and beef cows and calves were evaluated in Experiment III. Two studies consisted of two feeding strategies (Step-up vs. Flat-rate) and two diets (Control vs. Alternative). There were no differences between treatments in the cow LW, body condition score (BCS), calf pre-weaning LW gain and cow reproduction. A flat-rate strategy can be practised in the nutrition of mature suckler cows. Oat hull based flour-mill by product can partly replace grass silage and straw in the winter diet. Whole-crop barley silage can be offered as a sole feed to suckler cows. Experiment IV evaluated during the winter feeding period the effects of replacing grass silage with whole-crop barley or oat silage on mature beef cow and calf performance. Both whole-crop silages were suitable winter feeds for suckler cows in cold outdoor winter conditions. Experiment V aimed at assessing the effects of daily feeding vs. feeding every third day on the performance of mature beef cows and calves. No differences between the treatments were observed in cow LW, BCS, milk production and calf LW. The serum concentrations of urea and long-chain fatty acids were increased on the third day after feeding in the cows fed every third day. Despite of that the feeding every third day is an acceptable feeding strategy for mature suckler cows. Experiment VI studied the effects of feeding levels and long-term cold climatic conditions on mature beef cows and calves. The cows were overwintered in outdoor facilities or in an uninsulated indoor facility. Whole-crop barley silage was offered either ad libitum or restricted. All the facilities offered adequate shelter for the cows. The restricted offering of whole-crop barley silage provided enough energy for the cows. The Finnish energy recommendations for dairy cows were too high for mature beef breed suckler cows in good body condition at housing, even in cold conditions. Therefore, there is need to determine feeding recommendations for suckler cows in Finland. The results showed that the required amount of energy can be offered to the cows using conventional or alternative feeds provided at a lower feeding level, with an inaccurate feeding, flat-rate feeding or feeding every third day strategy. The cows must have an opportunity to replenish the LW and BCS losses at pasture before the next winter. Production in cold conditions can be practised in inexpensive facilities when shelter against rain and wind, a dry resting place, adequate amounts of feed suitable for cold conditions and water are provided for the animals as was done in the present study.

Digestion capasity, nutrient digestibilities and physico-chemical conditions in the intestine influenced by the age of growing turkeys

Six experiments have been conducted to examine digestibility and feeding value of domestic Finnish fibre-rich cereals (barley and oats as compared to maize and wheat) and protein sources (rapeseed meal and cake, peas, faba beans, lupin seeds) for growing turkeys and to investigate effects of age of the birds (from 3 to 12 weeks of age) on digestion process and estimated nutrient digestibility and energy values. Besides, an objective of the study was to test applications of digestibility research methodology for turkeys. Total tract digestibility and apparent metabolizable energy (AME) was assayed in experimental cages using excreta collection, and a slaughter method was applied to sample small intestinal digesta for determination of apparent ileal crude protein digestibility (AICPD), jejuno-duodenal digesta viscosity and caecal volatile fatty acid (VFA) concentration. Digesta viscosity decreased and caecal VFA production increased with age of growing turkeys. Digesta retention times in the small intestine were generally longer in the older birds than in the younger ones. Crude fat digestibility and AME increased with age of growing turkeys, especially with viscous diets. AICPD seemed to decrease with age in most cases. Supplementation with β-gucanase-xylanase decreased viscosity, improved crude fat digestibility and metabolizable energy value and increased VFA production especially in barley-fed turkeys and especially in the young birds. Poor protein digestibility and low energy value of rapeseed meal and rapeseed cake decreased their feeding value for turkeys. In addition, a typical goitrogenic effect of rapeseed feeding was detected. Use of legume seeds as feed for growing turkeys is limited mostly by the low energy value in lupin seeds and the low ileal protein and amino acid digestibility in faba beans. Digestibility of fibre-rich protein sources was not improved with age of the turkeys. Euthanizing the turkeys for AICPD determination by carbon dioxide and bleeding led to lower digestibility values than mechanical stunning and cervical dislocation, suggesting inferiority of carbon dioxide stunning in experimental use. Comparison of AICPD and AME results obtained using different markers showed that considerable differences may occur, especially on total tract level, when acid-insoluble ash gave considerably lower AME values than titanium dioxide and chromic oxide.

Bacillus cereus spores and cereulide in food-borne illness

B. cereus is a gram-positive bacterium that possesses two different forms of life:the large, rod-shaped cells (ca. 0.002 mm by 0.004 mm) that are able to propagate and the small (0.001 mm), oval shaped spores. The spores can survive in almost any environment for up to centuries without nourishment or water. They are insensitive towards most agents that normally kill bacteria: heating up to several hours at 90 ºC, radiation, disinfectants and extreme alkaline (≥ pH 13) and acid (≤ pH 1) environment. The spores are highly hydrophobic and therefore make them tend to stick to all kinds of surfaces, steel, plastics and live cells. In favorable conditions the spores of B. cereus may germinate into vegetative cells capable of producing food poisoning toxins. The toxins can be heat-labile protein formed after ingestion of the contaminated food, inside the gastrointestinal tract (diarrhoeal toxins), or heat stable peptides formed in the food (emesis causing toxin, cereulide). Cereulide cannot be inactivated in foods by cooking or any other procedure applicable on food. Cereulide in consumed food causes serious illness in human, even fatalities. In this thesis, B. cereus strains originating from different kinds of foods and environments and 8 different countries were inspected for their capability of forming cereulide. Of the 1041 isolates from soil, animal feed, water, air, used bedding, grass, dung and equipment only 1.2 % were capable of producing cereulide, whereas of the 144 isolates originating from foods 24 % were cereulide producers. Cereulide was detected by two methods: by its toxicity towards mammalian cells (sperm assay) and by its peculiar chemical structure using liquid-chromatograph-mass spectrometry equipment. B. cereus is known as one of the most frequent bacteria occurring in food. Most foods contain more than one kind of B. cereus. When randomly selected 100 isolates of B. cereus from commercial infant foods (dry formulas) were tested, 11% of these produced cereulide. Considering a frequent content of 103 to 104 cfu (colony forming units) of B. cereus per gram of infant food formula (dry), it appears likely that most servings (200 ml, 30 g of the powder reconstituted with water) may contain cereulide producers. When a reconstituted infant formula was inoculated with >105 cfu of cereulide producing B. cereus per ml and left at room temperature, cereulide accumulated to food poisoning levels (> 0.1 mg of cereulide per serving) within 24 hours. Paradoxically, the amount of cereulide (per g of food) increased 10 to 50 fold when the food was diluted 4 - 15 fold with water. The amount of the produced cereulide strongly depended on the composition of the formula: most toxin was formed in formulas with cereals mixed with milk, and least toxin in formulas based on milk only. In spite of the aggressive cleaning practices executed by the modern dairy industry, certain genotypes of B. cereus appear to colonise the silos tanks. In this thesis four strategies to explain their survival of their spores in dairy silos were identified. First, high survival (log 15 min kill ≤ 1.5) in the hot alkaline (pH >13) wash liquid, used at the dairies for cleaning-in-place. Second, efficient adherence of the spores to stainless steel from cold water. Third, a cereulide producing group with spores characterized by slow germination in rich medium and well preserved viability when exposed to heating at 90 ºC. Fourth, spores capable of germinating at 8 ºC and possessing the psychrotolerance gene, cspA. There were indications that spores highly resistant to hot 1% sodium hydroxide may be effectively inactivated by hot 0.9% nitric acid. Eight out of the 14 dairy silo tank isolates possessing hot alkali resistant spores were capable of germinating and forming biofilm in whole milk, not previously reported for B. cereus. In this thesis it was shown that cereulide producing B. cereus was capable of inhibiting the growth of cereulide non-producing B. cereus occurring in the same food. This phenomenon, called antagonism, has long been known to exist between B. cereus and other microbial species, e.g. various species of Bacillus, gram-negative bacteria and plant pathogenic fungi. In this thesis intra-species antagonism of B. cereus was shown for the first time. This brother-killing did not depend on the cereulide molecule, also some of the cereulide non-producers were potent antagonists. Interestingly, the antagonistic clades were most frequently found in isolates from food implicated with human illness. The antagonistic property was therefore proposed in this thesis as a novel virulence factor that increases the human morbidity of the species B. cereus, in particular of the cereulide producers.

Lactic acid bacteria and their antimicrobial peptides : Induction,detection,partial characterization, and potential applications

Bacteriocin-producing lactic acid bacteria and their isolated peptide bacteriocins are of value to control pathogens and spoiling microorganisms in foods and feed. Nisin is the only bacteriocin that is commonly accepted as a food preservative and has a broad spectrum of activity against Gram-positive organisms including spore forming bacteria. In this study nisin induction was studied from two perspectives, induction from inside of the cell and selection of nisin inducible strains with increased nisin induction sensitivity. The results showed that a mutation in the nisin precursor transporter NisT rendered L. lactis incapable of nisin secretion and lead to nisin accumulation inside the cells. Intracellular proteolytic activity could cleave the N-terminal leader peptide of nisin precursor, resulting in active nisin in the cells. Using a nisin sensitive GFP bioassay it could be shown, that the active intracellular nisin could function as an inducer without any detectable release from the cells. The results suggested that nisin can be inserted into the cytoplasmic membrane from inside the cell and activate NisK. This model of two-component regulation may be a general mechanism of how amphiphilic signals activate the histidine kinase sensor and would represent a novel way for a signal transduction pathway to recognize its signal. In addition, nisin induction was studied through the isolation of natural mutants of the GFPuv nisin bioassay strain L. lactis LAC275 using fl uorescence-activated cell sorting (FACS). The isolated mutant strains represent second generation of GFPuv bioassay strains which can allow the detection of nisin at lower levels. The applied aspect of this thesis was focused on the potential of bacteriocins in chicken farming. One aim was to study nisin as a potential growth promoter in chicken feed. Therefore, the lactic acid bacteria of chicken crop and the nisin sensitivity of the isolated strains were tested. It was found that in the crop Lactobacillus reuteri, L. salivarius and L. crispatus were the dominating bacteria and variation in nisin resistance level of these strains was found. This suggested that nisin may be used as growth promoter without wiping out the dominating bacterial species in the crop. As the isolated lactobacilli may serve as bacteria promoting chicken health or reducing zoonoosis and bacteriocin production is one property associated with probiotics, the isolated strains were screened for bacteriocin activity against the pathogen Campylobacter jejuni. The results showed that many of the isolated L. salivarius strains could inhibit the growth of C. jejuni. The bacteriocin of the L. salivarius LAB47 strain, with the strongest activity, was further characterized. Salivaricin 47 is heat-stable and active in pH range 3 to 8, and the molecular mass was estimated to be approximately 3.2 kDa based on tricine SDS-PAGE analysis.

A dynamic programming model for optimising feeding and slaughter decisions regarding fattening pigs

Costs of purchasing new piglets and of feeding them until slaughter are the main variable expenditures in pig fattening. They both depend on slaughter intensity, the nature of feeding patterns and the technological constraints of pig fattening, such as genotype. Therefore, it is of interest to examine the effect of production technology and changes in input and output prices on feeding and slaughter decisions. This study examines the problem by using a dynamic programming model that links genetic characteristics of a pig to feeding decisions and the timing of slaughter and takes into account how these jointly affect the quality-adjusted value of a carcass. The model simulates the growth mechanism of a pig under optional feeding and slaughter patterns and then solves the optimal feeding and slaughter decisions recursively. The state of nature and the genotype of a pig are known in the analysis. The main contribution of this study is the dynamic approach that explicitly takes into account carcass quality while simultaneously optimising feeding and slaughter decisions. The method maximises the internal rate of return to the capacity unit. Hence, the results can have vital impact on competitiveness of pig production, which is known to be quite capital-intensive. The results suggest that producer can significantly benefit from improvements in the pig's genotype, because they improve efficiency of pig production. The annual benefits from obtaining pigs of improved genotype can be more than €20 per capacity unit. The annual net benefits of animal breeding to pig farms can also be considerable. Animals of improved genotype can reach optimal slaughter maturity quicker and produce leaner meat than animals of poor genotype. In order to fully utilise the benefits of animal breeding, the producer must adjust feeding and slaughter patterns on the basis of genotype. The results suggest that the producer can benefit from flexible feeding technology. The flexible feeding technology segregates pigs into groups according to their weight, carcass leanness, genotype and sex and thereafter optimises feeding and slaughter decisions separately for these groups. Typically, such a technology provides incentives to feed piglets with protein-rich feed such that the genetic potential to produce leaner meat is fully utilised. When the pig approaches slaughter maturity, the share of protein-rich feed in the diet gradually decreases and the amount of energy-rich feed increases. Generally, the optimal slaughter weight is within the weight range that pays the highest price per kilogram of pig meat. The optimal feeding pattern and the optimal timing of slaughter depend on price ratios. Particularly, an increase in the price of pig meat provides incentives to increase the growth rates up to the pig's biological maximum by increasing the amount of energy in the feed. Price changes and changes in slaughter premium can also have large income effects. Key words: barley, carcass composition, dynamic programming, feeding, genotypes, lean, pig fattening, precision agriculture, productivity, slaughter weight, soybeans

Liikennepolttoaineeksi tarkoitettu ohraetanoli, Suomen ohramarkkinat ja ilmastonmuutos

The aim of the thesis was to analyze the use of barley as an input for bioethanol production and the impacts the use has on the Finnish barley markets. Two main research questions were formulated. First, privately and socially optimal bioethanol production levels were examined. In the social optimum, the climate benefits of bioethanol production were considered. It was calculated that the production and use of bioethanol created smaller CO2 -emissions when compared with the production and use of gasoline. Second, the impacts of bioethanol production on farmland allocation and agricultural production were analyzed. In more detail, the second aim was to analyze the farmland allocation between wheat and barley cultivation and green set aside in the private and social optimum. An analytical model was produced to analyze the barley markets in Finland. To provide an empirical counterpart to this model, existing research data on bioethanol production and barley cultivation was used. The aim of the model was to analyze the supply and the demand as well as market equilibrium of barley. Furthermore, the model provided a framework for analyzing the differences between the private optimum and social optimum of bioethanol production in Finland. The demand for barley consists of animal feed demand and bioethanol demand. On the supply side, a heterogeneous model of farmland quality was used. With this framework, it is possible to analyze farmland allocation between barley and wheat cultivation and green set aside and how the climate benefits of bioethanol production affects the allocation. Moreover, the relative changes in barley price between the private and social optimum were analyzed. Based on the empirical analysis, the private optimum for barley based bioethanol production is 58 691 metric tons. However, the social optimum for barley based bioethanol production is 72 736 metric tons. The portion of farmland that is allocated to barley cultivation is increased if the climate benefits of bioethanol production are considered. In the private optimum, 1/19 of the total farmland is allocated to barley cultivation whereas in social optimum the share increases to 7/19. Furthermore, the increase in barley price between private and social optimum is rather modest. Total increase in price is only about 1,8 percent.