Browsing by Subject "aminohappo"

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  • Lahtinen, Sonja (Helsingin yliopisto, 2020)
    The purpose of the study was to determine the possibilities of utilizing the by-product streams of fish processing industry in the production of fish oil and silage. In this thesis the effect of different antioxidants on the preservation of salmon scraps was studied and the utilisation of the dregs from which the oil was extracted as feed was considered. The scarps preserved in this study were from farmed salmon and consisted of internal organs and roe. The study had two experiments. The first one lasted for 65 days and in it formic acid was added to the scraps for a final acid content of 3 %. The scraps with only the formic acid acted as the control and the four different antioxidants studied were butylated hydroxyanisole (BHA), tert-butylhydroquinone (TBHQ), ethoxyquin mixture (Helmlox) and butylated hydroxytoluene (Ionol). The amount of Ionol was 100 ppm and for BHA, TBHQ and Helmlox there were two different amounts, 50 and 100 ppm. The second experiment lasted for 29 days and the formic acid content was 1 %. BHA and Helmlox antioxidants were used with an amount of 10 ppm. The oil and sediment parts from the experiments were analyzed separately. Based on the analyses the antioxidants or their amounts did not have an effect on the values extracted from the sediment. In the first experiment the analyses from the oil gave mostly highly significant (p<0,001) differences between the control and the antioxidants. There were also clear differences between the antioxidants but the different amounts did not have an effect on the AV-, PV- and TOTOX-values. In the second experiment the analyses from the oil gave only minor differences between the control and the antioxidants, the differences between different antioxidants being larger. The peroxide value of BHA was clearly larger than that of the control and Helmlox and therefore also the TOTOX-value was larger, which contradicts the results of the first experiment. In the second experiment also the measured amino acid contents diminished when the antioxidants were added which contradicts previous research. TBHQ worked best as an antioxidant, then BHA, Ionol and Helmlox in order. The usage of the dregs from the scraps as feed for pigs and chicken is limited by its high fish oil content. However for fur animals salmon scraps are suitable as feed products if the amino acid deficiencies are compensated with other feed products.
  • Marttinen, Eeva (Helsingfors universitet, 2010)
    Nitrogen is usually the growth limiting nutrient in boreal forest soils. Most of the nitrogen is bound to organic fraction, and low bioavailability of nitrogen delimits plant growth in boreal forest soils. Amino acids are easily available nitrogen compounds and thus they are important nitrogen sources for soil microorganisms. Almost all boreal forest trees form mycorrhizal assoociations with fungi. Mycorrhizal fungi produce wide variety of enzymes which break down organic nitrogen compounds. So far there is little knowledge of amino acid mineralization mechanisms of ectomycorrhizal fungi. L-amino acid oxidase (LAO) catalyses the mineralization of amino acids to ammonium. The ectomycorrhizal fungi Hebeloma spp. and Laccaria spp. have been shown to possess LAO enzyme activities. It has been proposed that LAO is one of the nitrogen mineralization mechanisms in ectomycorrhizal fungi, but so far no LAO genes have been described from basidiomycete fungi. In this study the first LAO gene sequences from the basidiomycete fungus Hebeloma cylindrosporum was described. The RACE-PCR -method was used to determine the 3´ and 5´ end sequences of the cDNA of the LAO1 gene. Based on the obtained sequences, primers to isolate the genomic DNA and cDNA sequences of the LAO1 gene were designed. The structure of the LAO1 gene, which is composed of five exons and four introns, was determined. Binding site of nitrogen regulating protein was found from upstream region of LAO1-gene. The partial genomic DNA sequence of gene adjacent to LAO1-gene was also measured. In the L. bicolor genome the gene preceding the LAO1 gene has been annotated as a putative pyruvate decarboxylase. In this study the partial cDNA sequence of another LAO-homolog of H. cylindrosporum was also determined. The LAO gene from another basidiomycete fungus, Laccaria bicolor, was also recognised. The gene model of LAO gene of L. bicolor was unannotated in the NCBI database. Based on the phylogenetic tree of LAO-related protein sequences, the ancestral form of LAO gene has been duplicated. This study provides molecular biological information on the catabolic mechanisms of amino acids in ectomycorrhizal fungi. Ammonium ions, produced by ectomycorrhizal fungi, might be a significant source of nitrogen for plants and other soil microbes. It is possible that LAO is an important factor of nitrogen cycle in soils of boreal forests.
  • Termonen, Tytti (Helsingin yliopisto, 2015)
    Tämän maisterintutkielman tavoitteena oli selvittää härkäpapu-vehnäsäilörehun vaikutus lypsylehmän maitotuotokseen ja aineenvaihduntaan, kun puolet nurmisäilörehun kuiva-aineesta korvataan härkäpapu-kevätvehnäsäilörehulla. Lisäksi tavoitteena oli selvittää väkirehun valkuaistason vaikutus lisättäessä rypsirehun määrää ruokinnassa. Koe suoritettiin Viikin opetus- ja tutkimustilan navetassa keväällä 2014. Kokeessa oli mukana kahdeksan vähintään kaksi kertaa poikinutta ay-lehmää, joiden poikimisesta oli kulunut kokeen alkaessa keskimäärin 100 päivää. Koemallina oli rinnakkain toistettu 4x4 latinalainen neliö. Koekäsittelyinä olivat nurmisäilörehu (D-arvo 678 g/kg ka) täydennettynä väkirehulla, jossa oli raakavalkuaista joko 175 g/kg ka tai 200 g/kg ka, sekä härkäpapu-vehnäsäilörehun ja nurmisäilörehun seos (1:1) (D-arvo 642 g/kg ka) täydennettynä edellä mainituilla väkirehun valkuaistasoilla. Molemmat säilörehut olivat käymislaadultaan hyviä. Syönnissä ja maitotuotoksessa ei havaittu merkitsevää eroa säilörehujen tai väkirehun valkuaistasojen välillä. Kuidun sulavuudessa havaittiin yhdysvaikutus säilörehun kasvilajikoostumuksen ja väkirehun valkuaispitoisuuden välillä. Härkäpapu-vehnäsäilörehua sisältävällä ruokinnalla kuidun sulavuus parani rypsimäärän lisääntyessä, mutta nurmisäilörehuruokinnalla sulavuus heikkeni. Kuidun sulavuus oli parempi nurmisäilörehuruokinnalla kuin härkäpapu-vehnäsäilörehua sisältävällä ruokinnalla. Väkirehun valkuaispitoisuuden suurentuessa maidon rasvapitoisuus ja rasvatuotos pienenivät, jolloin myös energiakorjattu maitotuotos pieneni. Typen hyväksikäyttö maidontuotantoon heikkeni väkirehun valkuaispitoisuuden suurentuessa. Plasman vapaiden rasvahappojen ja haaraketjuisten aminohappojen pitoisuus plasmassa oli merkitsevästi suurempi härkäpapu-vehnäsäilörehua sisältävillä ruokinnoilla kuin nurmisäilörehuruokinnoilla. Plasman insuliinipitoisuus suureni väkirehun valkuaispitoisuuden suurentuessa molemmilla säilörehuruokinnoilla. Tulosten perusteella ensimmäisen sadon nurmisäilörehun korvaaminen härkäpapu-vehnäsäilörehulla ei aiheuta tuotannon heikkenemistä, kun nurmisäilörehusta korvataan puolet härkäpapu-vehnäsäilörehulla. Siten härkäpapu-vehnäsäilörehu osana karkearehua sopii hyvin lypsylehmien ruokintaan. Väkirehun raakavalkuaispitoisuuden lisääminen 175 g:sta/kg ka 200 g:n/kg ka ei lisännyt maitotuotosta, mutta vähensi maidon rasvapitoisuutta molempia säilörehuja käytettäessä.
  • Virtanen, Katri (Helsingfors universitet, 2014)
    The aim of this study was to examine how immunocastration effects on pigs growth rate, feed efficiency and carcass traits and how high amounts of ileal digestible amino acids can pigs utilize. Another aim of this study was to examine if immunocastrating causes pain or stress to pigs in short period of time. This experiment was carried out in research piggery of MTT Hyvinkää. Pigs were splitted in six test groups by their gender (immunocastrated and surgically castrated males) and the level of standardized ileal digestible amino acids (low level, recommended level and high level). Growth and feed efficiency were measured by weighting pigs in the beginning of first growing phase, during growing weeks 6 and 9 and at the end of the trial. Carcass traits and meat quality were measured immediately after slaughter by Autofom-equipment. Pigs were videotaped for three days during each injection and their behavior was recorded. Statistical analysis of growth features and carcass traits were accomplished by SAS 9.3 GLM-procedure and behavioral changes by SPSS-software. The level of ileal digestible amino acids improved average daily gain (ADG) on both genders. Surgically castrated males achieved their best ADG during growing weeks 6-8 while immunocas trated males achieved their best results between growing week 9 and slaughter and at that time their results were best of all test groups. Increase of ileal digestible amino acids improved feed efficiency in all test groups. Stress indicating behavioral changes like standing, being alone and changes in activity, was noticed especially during first injection at the same day the injection was given. Second injection increased behavioral changes more the day after injection. Based on these results it seems that immunocastrating causes stress to pigs but it lasts only for a short time.
  • Laukkanen, Salla (Helsingfors universitet, 2017)
    Microalgae are unicellular organisms with excellent nutritional composition, ability to efficiently produce biomass and low environmental demands. The use of microalgae in animal feeds is common in aquaculture and newly introduced to animal husbandry. There is a growing need for alternative protein feeds to diminish the environmental cost of feed production and competition with food production. This research examined if soy protein can be replaced with microalgal protein in concentrate feeding of dairy cows. The effect of protein source on feed intake, milk production, milk composition, amino acid intake and use in mammary gland as well as plasma metabolites were evaluated. The feeding experiment was conducted in the research farm of the University of Helsinki in the summer of 2014. The study design was a 4x4 Latin square with four multiparous ayrshire dairy cows and four different experimental diets (isonitrogenously soybean meal (Glycine max), Spirulina platensis, Chlorella vulgaris or 1:1 mixture of Chlorella and Nannochloropsis gaditana as protein feed). The experimental concentrates (12.5 kg/d) were based on cereals and molassed sugarbeet pulp. The cows were given grass silage ad libitum. The physiological feeding experiment lasted for 12 weeks, with four experimental periods of three weeks. Feed intake was recorded and samples of feed, milk, feces and blood were taken to determine the effect of the experimental feeds on the cows. Inclusion of microalgae lowered the intake of concentrate feeds, but overall dry matter intake remained unchanged as the intake of grass silage was increased. The only effect on milk production and milk composition was the slightly higher fat concentration of milk when microalgal feeds were fed. In plasma, acetic acid and free fatty acid concentrations were higher and insulin concentrations lower when feed included microalgae, and also the mammary metabolism of these metabolites was affected by the experimental diets. The results refer to slight changes in rumen fermentation and mammary gland metabolism when microalgae replaced soy in the feeds. The effects of different feeds on amino acid metabolism were minor. Based on mammary uptake-output ratio, the most limiting amino acid in all diets seemed to be methionine. Based on the results of this experiment, microalgal feeds are equal or even slightly superior to soy as a protein feed of dairy cows when it comes to nutritional composition and productive responses. Inferior palatability of microalgae compared to soy, high production costs of microalgal feeds and lack of systematic scientific research are nevertheless hindering the large-scale commercial use of microalgae in domestic animal feeds.
  • Ketonen, Krista (Helsingfors universitet, 2014)
    Variation of the protein and amino acid content of barley, wheat and oats were studied. Diets based on grain samples of different protein content were optimized for pigs and poultry.The study went on to optimize diets for pigs and poultry with grains of different protein contents. The amino acid and raw protein analysis was undertaken on 38 grain samples. Correlations were calculated between different variables in grain samples and linear regression analysis was conducted between the protein and amino acid composition. The best estimate for amino acid concentrations of cereals was the protein content. The relative content of amino acids decreases as protein content increases and especially so in barley and wheat. Most reliable regression equations between amino acid and protein content were made for barley and wheat samples. For oat reliable regression equations could not be made. Oats also differed by other features from barley and wheat as it correlated with different variables compared to barley and wheat. Amount of needed protein concentrate levels decreased when barley and wheat protein and amino acid contents were considered in optimization. Protein concentrate levels did not decrease when used oat sample with highest protein content.
  • Hassan, Ghada (Helsingfors universitet, 2016)
    The aim of this study was to synthesize antimicrobial and anti-biofilm agents using abietic (AA) and dehydroabietic acids (DHAA). Bacterial biofilms are formed when bacteria cells cluster together within a self-produced extracellular matrix. This lifestyle makes bacteria highly resilient to different environmental stresses and conventional antibiotics when compared to single-cell bacteria. Currently, there are no approved anti-biofilm agents as drugs and only a few number of compounds can selectively target biofilms and eradicate them at low concentrations. Potent drugs targeting them are needed. AA and DHAA are abietane-type diterpenoids found in the resin of conifer trees. Antibacterial effects of resin acids have been widely studied, specifically against methicillin-resistant Staphylococcus aureus strain (MRSA). Through the combination of DHAA with different amino acids, Manner et al. (2015) discovered a new class of hybrid compounds that target both planktonic and biofilm bacteria in Staphylococcus aureus. The study group also discovered two of the most potent abietane-type anti-biofilm agents reported so far in literature. This thesis followed the work of the research group by designing and synthesizing additional AA and DHAA derivatives to target bacterial biofilms. Rings A, B and C of the diterpenoid core were modified and 24 derivatives were successfully synthesized. Amino acids were attached to the compounds either before or after ring modification. Standard structural elucidation techniques were used to confirm the structure of the synthesized compounds.
  • Päkkilä, Liisa-Maria (Helsingfors universitet, 2016)
    The aim of this study was to investigate effect of faba bean on amino acid, glucose, NEFA, BHBA, acetate and insulin concentration in plasma. In addition, AV-difference, extraction, and uptake of nutrients by mammary gland was investigated. The primary purpose of this study was to examine suitability of faba bean as protein feed for dairy cows. Firstly, faba bean was compared to grain based control feed, which did not contain any protein supplement, and it was fed with restrictively fermented silage. Secondly, faba bean was compared to rapeseed meal and mixture of rapeseed meal and faba bean as isonitrogenous and isoenergetic diets. Eight multiparous cows in the experiment were divided into two blocks and replicated 4 × 4 latin square design was used. Concentrates were fed at a rate of 14 kg/d and silage was fed ad libitum. Cows were approximately 100 days in milk in the beginning of the experiment. Rapeseed meal did not increase silage intake as much as expected according to results of earlier experiments. When rapeseed meal was replaced with faba bean, milk production, energy corrected milk production, protein production and fat production increased quadratically. When protein content of the concentrate increased, concentration of essential amino acids increased in plasma. When rapeseed meal was replaced with faba bean, concentration of non-essential amino acids increased linearly. Concentration of histidine in plasma was high on all treatments compared to results of earlier experiments. Concentration of methionine in plasma was at the same level as results of earlier experiments. Extraction of methionine was exceptionally high when rapeseed meal was fed, but uptake rate of methionine by mammary gland was bigger than output rate. Concentration of glucose in plasma was high compared to results of earlier experiments on every treatment. Relative and absolute uptake of glucose by mammary gland was especially high when mixed protein feed was fed. Supply of energy and amino acid profile corresponded well to cows needs with mixed protein feed.
  • Moslova, Karina (Helsingin yliopisto, 2020)
    Ribonukleiinihapot (RNA) toimivat kriittisissä biologisissa tehtävissä geneettisen tiedon välittäjinä solun sisällä. Sen lisäksi niillä on merkittävä rooli perintöaineksen säilymisessä ja siirtymisessä sekä katalyyttisten toimintojen ylläpitämisessä. RNA-modifikaatioiden on katsottu toimivan ylimääräisenä tiedonvälittäjäkerroksena solufysiologian säätelemisessä nukleotidisekvenssien koodauksessa. Tämän Pro gradun kirjallisuusosassa esitellään viimeisten viiden vuoden ajalta (2014-2019) vakiintuneet tekniikat sekä ohjelmistot RNA-modifikaatioiden tutkimuksissa. RNA-modifikaatioiden monimuotoisten biologisten toimintojen selvittäminen perustuu tarkkaan detektointiin, kvantitointiin, ja modifikaatioiden kartoitukseen. Näytteen esikäsittely mm. kemiallinen leimaaminen on tärkeässä roolissa onnistuneen analyysin kannalta. Työssä kerrotaan mm. massaspektrometrisistä (MS) identifioinneista ja rakennetutkimuksista sekä sekvenssointtitekniikoiden hyödyntämisestä. Useiden viime vuosien aikana menetelmien kehittämisessä on edistytty merkittävästi. Erityisesti MS datan käsittelyyn soveltuvia ohjelmistoja on luotu tandemmassaspektrien tulkitsemiseksi. Massaspektrien käsittelyn monipuolistaminen on johtanut uusiin lähestymistapoihin. joista voidaan saada kvalitatiivista ja kvantitatiivista tietoa RNA-modifikaatioista, usein jopa sekvenssispesifisyyden tasolla. Koska RNA-tutkimus on vielä varhaisessa vaiheessa, se vaatii paljon tutkimustyötä. Uusien menetelmien, tekniikoiden ja ohjelmistojen kehittymisen myötä ymmärrys RNA-modifikaatioiden olemassaolosta, tarkoituksesta, toiminnasta ja vaikutuksesta elämään kasvaa. Työn kokeellisen osion aiheena oli derivatisoida lähettiribonukleiinihappo (mRNA) fluorisoivaksi ja kehittää analyyttinen erotusmenetelmä kapillaarielektroforeesilla (CE) laserindusoitua fluoresenssidetektoria (LIF) käyttäen. Fluoreskeiini-5-isotiosyanaatti (FITC) valittiin derivatisointireagenssiksi sen monipuolisen käytettävyyden vuoksi ja helpon saatavuuden takia. Edellisestä huolimatta on yleisesti tiedossa FITC:n nopea hajoaminen, mutta tähän asti ei ole raportoitu, kuinka nopeasti FITC todellisuudessa muuttuu reagointikyvyttömäksi. Menetelmän kehitysvaiheessa käytettiin RNA-tyyppisiä biologisia molekyylejä, kuten BSA ja aminohappoja. Työssä testattiin erilaisia puskureita eri pH-arvoilla ja pitoisuuksilla. Lisäksi optimoitiin injektiopaine ja -aika sekä virta käyttäen CE-laitteistoa varustettuna diodirividetektorilla (CE-UV). Koska työn päätavoitteena oli erotusmenetelmän kehittäminen, derivatisointireaktion toimivuuteen ei heti kiinnitetty tarpeeksi huomiota. Menetelmä optimoitiin leimatuille aminohapoille ja BSA:lle. Lopuksi siirryttiin viimeistelemään menetelmä CE-LIF:llä. Derivatisointituotteiden tunnistus tehtiin ESI-TOF-MS:lla. Siinä yhteydessä huomattiin FITC:n nopea hajoaminen. Monien haasteellisten kokeilujen jälkeen päädyttiin keskittyä FITC:n stabiiliuden selvittämiseen. FITC:ä myydään samalla cas-numerolla kahta eri konformeeria ja tämän lisäksi vielä eri cas-numerolla kahta isomeeria. Näillä kaikilla on hieman erilaiset fysikaaliset ominaisuudet, jotka selvitettiin tässä työssä. Tutkittiin FITC:n eri konformeerien stabiilisuutta erilaisissa liuottimissa ja kahdessa lämpötilassa kymmenen päivän ajan käyttäen ESI-TOF-MS-laitetta sekä positiivisella että negatiivisella polariteetilla. Tutkimuksessa standardoidulla derivatisointiproseduurilla testattiin FITC-kemikaalin avulla LIF-detektorin toimintaa. Kuitenkin mRNA-derivatisointiin vaaditaan kalliit leimauskitit, joita ei tässä vaiheessa ollut mahdollista hankkia. Pysyminen alkuperäisessä suunnitelmassa ja optimoida CE-LIF -metodi fluoresoivalla FITC-reagenssilla lähetti-RNA:n leimaamiseen ei aikataulullisesti onnistunut. Siten projekti painottui tämän kokeellisen työn osalta derivatisoinnin optimointiin ja FITC-kemikaalien kinetiikkaan. Sen vuoksi projektissa tehtiin perusteellinen pohjatyö tuleville tutkimuksille. Kokeellisesti onnistuttiin leimaamaan nukleotidiemäksiä, jotka ovat lähetti-RNA:n perusosia. Selvitettiin FITC:n hajoamisnopeus ja liuottimet, joita FITC-tutkimuksissa on käytetty ja joita tulee käyttää, jotta reaktio onnistuu. Leimaus onnistui p-kinoidille. Saatujen tulosten perusteella on mahdollista laskea FITC:n optimaalinen määrä onnistuneen reaktion kannalta. Lisäksi osoitettiin, että leimausreaktio on herkkä, kun käytetään optimaalista puskuria ja optimipitoisuuksia. Jatkotutkimuksissa myös FITC-liuoksen valmistamiseen pitää kiinnittää paljon huomiota.