Browsing by Subject "maitohappobakteeri"

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  • Koljonen, Paula (Helsingfors universitet, 2017)
    Aflatoxins are harmful compounds found in food and feed. The literature review of this thesis looked at aflatoxin M1 (AFM1) found in milk and its occurrence, significance and prevention methods using lactic acid bacteria (LAB). Furthermore, the assay methods of aflatoxins and the factors affecting the analysis were discussed. The aim of the experimental work was 1) to investigate the ability of five LAB strains to bind AFM1 in vitro and 2) to investigate how different matrixes affect the assay of AFM1 and to evaluate the feasibility of the selected ELISA kit for the study. In the first phase of the study the AFM1 concentrations of three different matrixes were analyzed using ELISA when known concentrations of the AFM1-standardsolution were added (0, 20 ja 40 ppt). In the second phase of the study the abilities of five LAB strains to bind AFM1 in UHT skimmed milk and MRS-broth were investigated. The bacterial suspensions were incubated at +32 °C for one hour, and the AFM1 concentration in the matrixes were 50, 15 and 10 ppt. The unbound AFM1 concentrations of the supernatants were analyzed from the samples using ELISA. The unbound AFM1 concentrations were converted to the proportional portion of the bound AFM1. In this study, statistically significant differences were observed in the abilities of the LAB to bind AFM1. The viable cells of strain B2 27 (Lb. plantarum-, pentosus- or paraplantarum) were the best binders of AFM1. They removed 43.7 % AFM1 in UHT skimmed milk, where the AFM1 concentration was 15 ppt. Unlike in previous studies viable cells bound AFM1 better than the heat-killed cells. The used ELISA kit was a sensitive method for analyzing low concentrations of AFM1, but at higher concentrations the assay results were inaccurate. The nonspecific interaction due to the components of the matrixes had to be taken into account when the results were reviewed. In the future, it may be possible to utilize the LAB strains, such as B2 27 for reducing AFM1 the concentration in milk and probably in other foodstuffs. There is a need to develop a practical application which can be used in the binding of AFM1 using lactic acid bacteria and thus reduce the bioavailability of AFM1.
  • Viljanen, Janne (Helsingfors universitet, 2012)
    Exohomopolysaccharides (HOPS) are polysaccharides that are composed solely of glucose or fructose monomers and which are classified according to the linkage type between the monomers (e.g., dextran ?-[1?6]). Several different lactic acid bacteria (LAB) strains produce HOPS from sucrose. It is known that maltose can influence the synthesis of HOPS which can lead to formation of oligosaccharides. In fermented foods, HOPS and oligosaccharides can have both technological impact and prebiotic properties. The aim of this thesis was to investigate the synthesis of HOPS and oligosaccharides in mMRS broths and in oat bran gruel. Lactobacillus reuteri FUA 3048, Lb. reuteri LTH 5448, Weissella cibaria 10M and W. confusa DSM 20194 were used as starter cultures. HOPS and oligosaccharide yields in mMRS broths were analysed by ethanol precipitation and acid hydrolysis. An enzyme assisted method was used in order to quantify linear dextran content in freeze-dried oat gruels. Oligosaccharide profiles were analysed with high performance anion exchange chromatography with pulsed amperometric detetection. In mMRS broths, which contained 25 g/l sucrose (S-mMRS), LAB strains synthesised exosaccharides at concentrations of 0.9–2.3 g/l. Addition of maltose (34 g/l) significantly increased exosaccharide yield. This can be explained by strong malto-oligosaccharide (e.g., panose) synthesis. W. cibaria 10M produced 0.94 g/l linear dextran in S-mMRS broth and at 0.74 g/l in maltose containing broth and W. confusa DSM 20194 synthesised 1.7 g/l linear dextran in both broths. In oat gruel matrix (34 g/l maltose) W. confusa DSM 20194 synthesised 4.6 g/100 g dextran in freeze-dried gruel. In lower maltose concentrations dextran yield increased. Oligosaccharide profiles indicated that Weissella strains and the Lb. reuteri FUA 3048 strain synthesised gluco-oligosaccharides and Lb. reuteri LTH 5448 strain produced erlose. The results of this thesis give further insight in to role of maltose in HOPS synthesis in food matrices.
  • Puhakka, Laura (Helsingfors universitet, 2011)
    Tämän maisterin tutkielman tarkoituksena oli testata eri maitohappobakteerien ja niiden annostustason vaikutusta säilörehun käymislaatuun ja aerobiseen stabiilisuuteen. Biologisilla säilöntäaineilla eli ympeillä säilöttyjen rehujen laatua verrattiin kontrolleina toimiviin painorehuun ja muurahaishapolla tehtyyn rehuun. Koerehut tehtiin Helsingin yliopiston Maataloustieteiden laitoksella 7.6.2010. Timotei (Phleum pratense) - nurminata (Festuca pratensis) kasvusto korjattiin tähkimisen alkuvaiheessa D-arvon ollessa 711 g/kg ka. Nurmikasvuston kuiva-ainepitoisuus oli 170 g/kg heti niiton jälkeen määritettäessä ja neljän tunnin esikuivauksen jälkeen 208 g/kg. Rehuraaka-aine jaettiin kuuteen erään, joihin lisättiin säilöntäaine. Säilöntäainekäsittelyt olivat: 1) ei säilöntäainetta (painorehu), 2) muurahaishappo (100 %:na 4 l/t), 3) Lactobacillus plantarum ja Pediococcus acidilactici 1x10? pmy/g sekä pektinaasi-, ksylanaasi- ja sellulaasientsyymi, 4) L. plantarum 1x10? pmy/g, 5) L. plantarum 1x10? pmy/g ja 6) L. plantarum ja L. buchneri 2x10? pmy/g. Rehut säilöttiin laboratoriosiiloissa kolmena rinnakkaisena. Laboratoriosiilojen lisäksi rehua säilöttiin jokaisesta säilöntäainekäsittelystä kuuteen rinnakkaiseen minisiiloon säilönnän alkuajan fermentaation ja rehujen pH:n muutoksen seuraamiseksi. Minisiiloista seurattiin kaasuntuotantoa 21 päivän ajan. Raaka-aineen koostumus ja rehujen säilönnällinen laatu sekä aerobinen stabiilisuus määritettiin. Säilörehujen kuiva-ainepitoisuus oli suhteellisen pieni (n. 210 g/kg), jotta voitiin testata biologisten säilöntäaineiden tehoa märässä rehussa. Kosteassa rehussa biologisten säilöntäaineiden toiminnan onnistuminen on haastavaa. Koska raaka-aineen sokeripitoisuus oli kuitenkin erittäin suuri (196 g/kg ka), onnistui ymppirehujen säilöntä hyvin. Kaikkien koerehujen pH oli alle 4:n. Painorehu ei täyttänyt hyvän rehun kriteerejä ammoniumtypen osalta. Muurahaishapporehu oli laadultaan tyydyttävää etikkahapon pitoisuuden osalta. Rehussa tapahtui epätyypillistä etanolikäymistä hiivojen toimesta, jonka johdosta rehussa muodostui suuri määrä kaasua säilönnän aikana. Muurahaishapporehu ei kuitenkaan lämmennyt aerobisen stabiilisuuden mittauksen aikana, johtuen todennäköisesti suuresta etikkahapon määrästä ja toisaalta pienestä sokerin määrästä. Maitohappobakteerisäilöntäaineilla saatiin käymislaadultaan parempaa säilörehua verrattuna painorehuun, lukuun ottamatta maitohappobakteeri-entsyymirehua. Entsyymilisäyksestä ei todennäköisesti ollut hyötyä rehun säilöntälaadun kannalta. Raaka-aineen suuren sokeripitoisuuden johdosta säilörehussa oli koko säilönnän ajan tarpeeksi sokeria maitohappobakteerien käytettäväksi ja sokeri toimi siten substraattina haitallisille mikrobeille tuottaen suuren etikkahappopitoisuuden. Maitohappobakteeri-entsyymiseoksella tehty rehu oli etikkahappopitoisuuden perusteella heikkolaatuista. Lactobacillus plantarumin molemmilla annostustasoilla (1x10? ja 1x10? pmy/g rehua) saatiin laadultaan hyvää rehua. Suuri sokeripitoisuus molemmissa rehuissa johtui todennäköisesti raaka-aineen tavallista suuremmasta sokerin määrästä. Molempien rehujen maitohappo-etikkahappo-suhde oli melko korkea, viitaten homofermentatiiviseen maitohappokäymiseen. Lactobacillus buchneri –lisäyksellä rehu oli säilönnälliseltä laadultaan hyvää. Heterofermentatiivisen ympin lisäys nosti tyypillisesti rehun pH:ta sekä pienensi maitohappoetikkahappo-suhdelukua verrattuna homofermentatiiviseen ymppiin. L. buchneri –lisäys paransi hieman säilörehun aerobista stabiilisuutta verrattuna homofermentatiivisella maitohappobakteerilla säilöttyyn rehuun, mutta tulos ei ollut tilastollisesti merkitsevä.
  • Viinamäki, Miia (Helsingfors universitet, 2015)
    Ruisleipä on keskeinen elintarvike suomalaisten ravitsemuksessa. Sen valmistuksessa käytetään yhä perinteistä juuritaikinaa eli raskia, joka sisältää maitohappobakteereja ja mahdollisesti hiivoja. Ruis sisältää runsaasti fruktaaneja ja lisäksi raskin fermentoinnin aikana muodostuu mannitolia. Fruktaani ja mannitoli imeytyvät huonosti ohutsuolesta, mutta suoliston bakteerit fermentoivat ne nopeasti. Nopeasti fermentoituvat substraatit saattavat aiheuttaa vatsaoireita jopa yli 10 % väestöstä. Opinnäytetyössä tutkittiin erään teollisen raskin fruktaani- ja mannitolimetaboliaa, raskimikrobistoa sekä raskimikrobiston fruktaaninhajottamiskykyä. Tutkimuskohteeksi valittiin BF-raski. BF-raskin fruktaanin, mannitolin ja pelkistettyjen sokerien pitoisuutta mitattiin raskituksen edetessä. Mittausten jälkeen käyttövalmiista BF-raskista eristettiin maitohappobakteerit ja niiden fruktaanin hajottamiskykyä testattiin kromogeenisillä maljoilla ja puhdasviljelmistä tehdyillä fermentaatioilla. Lisäksi valmistettiin BF-raskin kaltainen spontaani raski pienessä mittakaavassa ja mitattiin sen fruktaanipitoisuutta. Tutkimuksessa havaittiin, että BF-raskin raskitusprosessin aikana fruktaani ja prosessissa syntynyt mannitoli hävisivät. Yhdellä raskin maitohappobakteereista havaittiin olevan kyky kuluttaa tehokkaasti fruktaania ja geenit, jotka koodaavat solunulkoista fruktaanihydrolaasi-entsyymiä sekä FOS/fruktoosi-kuljetussysteemiä. Lisäksi siltä löytyivät mannitolin käyttöön tarvittavat geenit. BF-raskin kaltainen fruktaania kuluttava raskisysteemi saatiin aikaan myös spontaanisti pienemmässä mittakaavassa. Fruktaanivapaan ruisraskin avulla voidaan valmistaa niukasti fruktaania ja mannitolia sisältävää ruisleipää. Tällainen ’vatsaystävällinen’ ruisleipä voisi sopia perinteisestä ruisleivästä vatsaoireita saaville kuluttajille.
  • Ahonen, Susanna (Helsingin yliopisto, 2018)
    Bacteriocins are ribosomally synthesized antimicrobial proteins or peptides that inhibit growth of other bacteria. Bacteriocins can be divided into three classes based on their characteristics. Bacteriocins from the genus Lactococcus have only been found from the classes I and II. These two classes include small heat resistant peptides. Class III includes larger heat sensitive proteins. Bacteriocins have different ways to inhibit bacteria. They can either kill the bacteria or prevent them from multiplying. Due to their safe properties, bacteriocins can be used in food production to prevent the growth of harmful bacteria. In this Master’s thesis, antimicrobial compound produced by Lactococcus lactis LAC460 isolated from idli batter, was studied. Based on inhibition tests it was found that the LAC460 prevented the growth of some lactic acid bacteria by forming an inhibition zone on indicator plate. Uncommon to bacteriocins, the zone expanded during several weeks of incubation. This indicates that the antimicrobial compound can also kill inactive non-growing target cells. Based on the enzymatic tests the antimicrobial compound turned out to be a protein, probably a bacteriocin. The LAC460 bacteriocin lost its activity already at 53 °C, as well as both in low and high pH. This indicates that the compound was not a peptide but rather a bigger protein. The strain LAC460 produced more bacteriocin when the growth media was strongly buffered at least to pH 7. The bacteriocin was produced only at later growth phase. Based on the results from the ribosomal 16S rDNA sequencing, API-test and whole genome sequencing, LAC460 turned out to be a mixed culture of two bacteria, L. lactis and Enterococcus faecium. These bacteria were separated from each other with selective media and it was found out that Lactococcus was the actual producer of the bacteriocin. No obvious bacteriocin gene was found from the genome sequence, but the results suggest that the LAC460 bacteriocin belongs to class III bacteriolysins. As Lactococcus strains have not been found to produce class III bacteriocins, the antimicrobial protein characterized in this study seems to be a new bacteriocin.
  • Lamminen, Marjukka (Helsingfors universitet, 2014)
    The aim of this study was to investigate the esilability of white lupin (Lupinus albus) and spring wheat (Triticum aestivum) bi-crop when ensiled as a whole-crop. There were two plant mixture ratios and two growth stages in the study. Silages made with different additives were compared with untreated silages, which were as a control. The experimental silages were made at the research farm of the University of Helsinki in Viikki on August 13th and 27th, 2012. At the first growth stage (K1) wheat was at the beginning of dough stage and the pods of white lupin were green and the seeds filled 50 % of space between septa. At the second growth stage (K2) wheat was at the end of dough stage and the seeds filled 75 % of space between septa. After mowing plant species were separated and two ratios were formed. The first ratio comprised 1/3 white lupin and 2/3 wheat and the second 2/3 white lupin and 1/3 wheat of fresh weight. The additive treatments were: 1) without additive (untreated), 2) formic acid (4 l/t as a 100% acid), 3) the mixture of sodium nitrite (0,75 kg/t) and hexamine (0,5 kg/t) and 4) Lactobacillus plantarum 1x106 cfu/g. Silages were ensiled in the laboratory scale silos in triplicate. Silos were opened after 100 days of ensiling. The chemical composition of raw material at harvest and the fermentation characteristics and aerobic stability of silages were determined in the experiment. The dry matter content of plant mixtures was about 220 g/kg in the mixtures with higher proportions of lupin and about 300 g/kg in the mixtures with higher proportions of wheat. The preservation of silages were impeded by the low dry matter content and high buffering capacity of white lupin, the low sugar content of the mixtures and clostridial contamination. As the proportion of lupin increased the crude protein and sugar content of the mixture increased but the ensilability of the mixture declined. Preservation without additive produced low quality silages in all silage batches. High amounts of ammonia, butyric acid and other fermentation acids typical of bad silage fermentation were detected in the untreated silages. The fermentation quality was good in silages made with Lactobacillus plantarum at the first growth stage when the raw material was dryer and the sugar content of mixtures were higher. Based on the butyric acid and ammonia concentrations the use of Lactobacillus plantarum resulted in poor quality silages at the second growth stage. The dose of formic acid in this experiment wasn’t probably high enough in regard to the poor ensiling properties of raw material and clostridial contamination. High amounts of butyric acid were detected in all silages treated with formic acid. At the second growth stage the ammonia concentrations were also high. The most effective additive in the experiment was the mixture of sodium nitrite and hexamine resulting in good fermentation quality in all silage batches. No butyric acid was detected from the mixtures with higher proportions of wheat and amounts were extremely low also in the mixtures with higher proportions of lupin. Most of the silages were aerobically stable during 13 days of the period of measurement. Only a few silos (5 out of 48) were aerobically deteriorated and those silos were very unstable as the warming was noticed within 2 days. Most of the deteriorated silos (4) were treated with biological additive. Based on the present results the ensiling of white lupin-wheat whole-crop silage is difficult. The composition of raw material suggests that silage should be made from the mixture where the proportion of lupin is higher and cut at the earlier growth stage if the maximization of crude protein and sugar content is wanted. However, the ensilability of raw material is more difficult on the basis of dry matter content and buffering capacity. The yield is also lower than later in the growth stage. Silage additive should always be used when ensiling white lupin. The mixture of sodium nitrite and hexamine was the most effective additive in the study.
  • Antila, Emmi (Helsingin yliopisto, 2021)
    Listeria monocytogenes on maailmanlaajuisesti yksi merkittävimmistä elintarvikevälitteistä tautia aiheuttavista bakteereista. L. monocytogenes -bakteeria esiintyy erityisesti raaoissa liha- ja kalavalmisteissa, pastöroimattomissa maitotuotteissa sekä tuoreissa kasviksissa. L. monocytogenes on ongelma elintarvikkeiden lisäksi myös tuotantolaitoksissa, joiden pinnoille muodostamissaan biofilmeissä bakteeria voi esiintyä saneerauksesta huolimatta jopa vuosia. Leuconostoc gelidum -maitohappobakteerit ovat ympäristössä esiintyviä bakteereja, jotka ovat yleisiä kylmäsäilytettyjen suojakaasu- ja tyhjiöpakattujen elintarvikkeiden valtalajeja viimeisenä käyttöpäivänä. Jotkut L. gelidum -kannat tuottavat pieniä antimikrobisia peptidejä, bakteriosiineja, joilla on osoitettu olevan L. monocytogenes -bakteerin kasvua estävä vaikutus. Tämän työn tavoitteena oli tutkia bakteriosiinia tuottavan ja tuottamattoman L. gelidum -kannan vaikutusta L. monocytogenes -bakteerin kasvuun kylmäsäilytetyissä elintarvikkeissa tarkkaillen samalla kantojen läsnäolon vaikutusta elintarvikkeen happamuuteen. Tutkimuksen hypoteesina oli, että I) L. gelidum- kantojen läsnäololla on L. monocytogenes -bakteerin kasvua estävä vaikutus ja II) bakteriosiinia tuottavan kannan estovaikutus on voimakkaampi. Estovaikutusta tutkittiin elintarvikkeissa (6 ºC), joihin oli siirrostettu L. monocytogenes ATCC 7644ᵀ -kantaa. Elintarvikkeista määritettiin L. monocytogenes -pesäkemäärä (pmy/g) useassa aikapisteessä L. monocytogenes -kannan kasvaessa yksin, yhdessä bakteriosiinia tuottavan L. gelidum Vvan 9 -kannan kanssa ja yhdessä bakteriosiinia tuottamattoman L. gelidum LMG 18811ᵀ -kannan kanssa. Kokeen aikana seurattiin myös mahdollisia muutoksia elintarvikkeiden pH-arvossa. Tulosten perusteella molemmilla L. gelidum -kannoilla oli L. monocytogenes ATCC 7644ᵀ -kannan kasvua hidastava vaikutus. Bakteriosiinia tuottavan Vvan 9 -kannan estovaikutuksen havaittiin kuitenkin olevan huomattavasti voimakkaampi kuin bakteriosiinia tuottamattoman LMG 18811ᵀ -kannan estovaikutuksen. Tämä viittaisi siihen, että hypoteesin mukaisesti Vvan 9 -kannan tuottamat bakteriosiinin kaltaiset yhdisteet tehostivat kannan estovaikutusta. L. gelidum -kantojen läsnäolo laski pH-arvoa matriiseissa, joissa ei ollut runsasta luonnollista mikrobistoa. Työ osoittaa, että L. gelidum Vvan 9 -kanta ja sen tuottamat bakteriosiinit ovat potentiaalinen keino torjua L. monocytogenes -bakteerin kasvua tyhjiö- ja suojakaasupakatuissa kylmäsäilytetyissä elintarvikkeissa. Kyseinen kanta sekä muut bakteriosiineja tuottavat L. gelidum -kannat tarjoavatkin jatkossa mielenkiintoisen tutkimuskohteen.
  • Alasalmi, Krista (Helsingin yliopisto, 2019)
    Sourdough baking is a process used for thousands of years and it’s still used to this day. Sourdough is a mixture of flour and water that has been fermented with lactic acid bacteria (LAB) and yeasts and it’s used to leaven the bread. Some of sourdough LAB produce exopolysaccharides (EPS) from sucrose in their metabolism. Dextran, which consists of α-(1→6) linked glucosyl units is the most common EPS. EPS produced by LAB have been widely studied to have different positive impacts on bread texture. The aim of this thesis was to screen the sourdough samples for EPS producing LAB and to analyze the structures of those EPS as accurately as possible. In the experimental part LAB were isolated from the sourdough samples and grown on MRS agar containing sucrose. All EPS forming colonies were then isolated from the plates and purified to obtain pure strains. The EPS produced by the strains were hydrolyzed enzymatically by dextranase and glucosidase after which their monosaccharides and enzyme resistant oligosaccharides were analyzed by HPAEC-PAD. The structures of the EPS were also analyzed with NMR. 13 EPS producing strains were isolated from the sourdough samples. Based on the HPAEC-PAD results all samples were found to be dextran because the enzymes were able to hydrolyze them. From the enzyme resistant oligosaccharide chromatograms it was seen that there were four different chromatographic profiles so there were four different EPS structures. NMR results confirmed that all EPS were dextrans. The NMR results also confirmed that there were four different structures among the EPS samples. All EPS had α-(1→3) linked branches. Two samples also had α-(1→2) linked branches. This research gave information about the EPS production by the sourdough’s LAB and also the structures of those EPS.
  • Tenhovirta, Santeri (Helsingin yliopisto, 2019)
    In the thesis, the effects of six different lactic acid bacteria (LAB) species on properties of sour beer was studied. As the fermentation by LAB may yield unexpected results, and as the prediction of specific organoleptic compounds present in LAB-fermented sour beers has proven to be challenging, sensory and chemical properties of sour beers fermented with selected LAB was assessed. The main research question was “are there statistically significant differences between the different sensory properties of the sour beers produced with different LAB species?”. Use of Lactobacillus plantarum, L. rhamnosus, L. brevis, L. buchneri, L. delbrueckii and previ-ously unreported in sour beer brewing, L. alimentarius, was assessed. Fermentation was carried over approximately six weeks, during which the changes in viable cell density, pH, refractive index, organic acid and ethanol (using PDA/RI) and sugar compositions (using HPAEC-PAD) were assessed. Sensory evaluation was performed in two parts: Overall preference rank test with untrained panellists and descriptive analysis with trained panellists. Differences were found in the results of viable cell count, organic acid composition and descriptive sensory analysis. Increase in lactic, acetic and succinic acid and ethanol over the fermentation period was detected. Decrease in citric acid, maltose, glucose and sucrose was detected. Novel L. alimentarius yielded fastest growth rate, with encouraging results from sensory analysis. L. delbrueckii did not grow in the substrate media, allowing possible infection to take place. L. brevis and L. buchneri yielded acetic acid concentrations detectable in sensory and chemical analysis. Increased lactic acid concentration was associated with increased overall rank, raspberry aroma and apple and vinous flavours, with negative correlation to butyric, rancid and yeasty flavours and bitter aftertaste. Further studies are suggested to assess the effects of fermentation temperature, O2 availability, wort composition and buffering capacity on the results of the LAB fermentations.