Browsing by study line "ei opintosuuntaa"

The field of food control is currently facing challenges due to phenomena such as climate change, globalization of food, and scarcity of official control resources. One important approach in improving efficient control of the safety and quality of food chains is risk-based food control. In this study, data-driven approaches were utilized to provide insights into some of the factors that have affected Finnish food business operators’ (FBOs’) compliance with food safety requirements in recent years. Qlik Sense, a data analytics solution built on the concept of visual analytics, and Python, a programming language suitable for data analysis, were used to analyze food inspection data recorded by local food safety authorities. Interactive data visualizations built in Qlik Sense aimed in supporting open government data (OGD) policies and risk-based control of the Finnish food chain, and so far, received good feedback from end-users. Additional insights into FBOs’ food safety compliance were gained by further analyzing the inspection data in Python along with municipal data. A logistic regression model fit to a subset of the study data found multiple statistically significant predictor variables from both datasets, but its performance was weak. The factors that affected food safety compliance most significantly were the number of years an FBO had operated and the basis for conducting an inspection. Operating years showed a positive correlation with compliance, while a negative relationship was observed with a variety of unplanned inspections, especially when they were conducted based on food poisoning suspects, inspection requests, or some other forms of contact. Out of all inspected food sectors, the one that increased the odds of compliance the most was the food transportation sector. The results of the study advocate for the potential of data-driven approaches in improving risk-based food control, as they are an effective way to gain insights into factors affecting the safety and quality of complex food chains.

In instantization milk powders’ reconstitution properties are improved. Typical instantization processes are agglomeration, in which particle size and porosity are increased, and lecithination, in which powder particle surface is made less hydrophobic. Both instantization methods have some drawbacks: Agglomeration is not sufficient for instantizing high fat milk powders, while lecithination tends to decrease powder’s flowability. The aim of this study was to review suitability of agglomeration in instantizing milk powder (SMP) and whole milk powder (WMP). Powders were agglomerated on fluid bed agglomerator by using different binder liquids. The physical properties, like wettability, flowability and particle size, of the agglomerated powders were characterized and compared to the unprocessed raw materials and reference powders (commercial instantized milk powders). According to the results reconstitution properties were improved more when using other binder liquids than only water. The fastest wetting on agglomerated SMP was achieved with sugar binder liquids. However, sugar also impaired powders’ flowability. On agglomerated WMPs fastest wetting occurred with phospholipid fraction as a binder liquid. Future research could investigate usage of different binder liquids in combination and optimizing agglomeration parameters for each binder liquid.

Phage lysins are enzymes that degrade bacterial cell wall. A wild-type Lactococcus lactis strain LAC460 secretes three phage lysins, LysL, LysP, and LysT, encoded by three different prophages. Unlike common phage lysins, these enzymes do not break down the host's cell wall. Therefore, these lysins can attack other L. lactis strains and behave like bacteriocins, antimicrobial proteins. The binding of a phage lysin to bacterial cell wall requires a specific cell wall binding domain (CBD) in the lysin. However, nothing about the CBDs of LysL, LysP and LysT is known. This study aimed to determine the CBDs of these three lysins and the target specificity of the lysins. Putative CBD regions of the lysins were fused with green fluorescent protein (GFPuv). GFPuv-CBD-LysL and GFPuv-CBD-LysT were ligated into the pASG-IBA4 vector and cloned in Escherichia coli DH5α. After all, only the construction of the GFPuv-CBD- LysL was successful resulting in fluorescent transformants. To analyse the binding of GFPuv- CBD-LysL to cells of different L. lactis strains, the fusion proteins were mixed with the LysL sensitive L. lactis MG1614, LysL resistant L. lactis LM0230, and the LysL producing LAC460 cells. With fluorescence microscope it could be seen that the GFPuv-CBD-LysL decorated the cell surface of L. lactis MG1614 with green fluorescence, but LM0230 and LAC460 cells remained non-fluorescent. The fluorescence of the cells was also measured with a fluorometer, showing strong fluorescence from MG1614, but nothing from the other two strains. This showed that the fusion protein specifically bound to the MG1614 cell surface, but it did not bind to the LysL resistant strain LM0230 or the LysL producer LAC460 cell. In conclusion, the results demonstrate that the C-terminus of LysL contains a specific cell wall binding domain. In addition, the results provide an explanation for how LAC460 can secrete LysL without autolysis, as phage lysins not able to bind onto peptidoglycan are unable to lyse cells.

3-Chloro-1,2-propanediol (3-MCPD), 2-chloro-1,3-propanediol (2-MCPD) and 2,3-epoxy-1-propanol (glycidol) and their fatty acid esters are contaminants formed during processing fat containing foodstuffs at high temperatures. Mainly MCPD and glycidyl esters have been found to be formed in the deodorization process of oils, and in vegetable oils, such as palm oil, they have been measured at high concentrations. In accordance with the restrictions imposed by the European Commission, the levels of glycidyl esters must be especially monitored, as they have been identified as potentially carcinogenic compounds. The aim of the study was to introduce and validate a gas chromatographic analysis method for glycidyl esters and MCPD esters for the Customs Laboratory. The method was validated for two matrices: first for oils and then for powdered infant formulas. In addition, the success of the validation was examined by analyzing various oil samples previously received by the Customs Laboratory. The Customs Laboratory is also involved in the activities of the European Union Reference Laboratory, for which it was intended to participate in the reference measurement organized by the EU Reference Laboratory. The method for the determination of 3-MCPD, 2-MCPD and glycidyl esters in oils and infant formulas was carried out according to the guidelines of the European Union Reference Laboratory for Contaminants (EURL-PC). Determination of MCPD and glycidyl ester concentrations in oils and infant formulas included the following steps: fat extraction by liquid-liquid extraction (for infant formulas), addition of standards, solid-phase extraction, conversion of glycidyl esters to 3-MBPD esters, transesterification, neutralization, salting out, derivatization and analysis with gas chromatography-mass spectrometry system. Concentrations were determined using internal standard method. The method was validated for the following parameters: specificity, selectivity, limit of detection and quantitation, reproducibility, repeatability, trueness, linearity and working range, stability and measurement uncertainty. The analytical method developed for the determination of MCPD and glycidyl esters was successfully validated for oils and powdered infant formulas. The developed method proved to be specific and selective. The limit of determination was found to be 6.3 µg/kg, 1.3 µg/kg and 0.8 µg/kg for the oil matrix 3-MCPD, 2-MCPD and glycidyl esters. The limits of determination for the infant formula were 5.4 µg/kg, 3.0 µg/kg and 1.6 µg/kg for 3-MCPD, 2-MCPD and glycidyl esters. Recoveries for MCPD and glycidyl esters in the oil and powdered infant formulas were 83-105%. R2 for calibration lines were greater than 0.99, and the lines were linear over the entire measurement range of 2-1000 µg/kg. The relative standard deviation of repeatability and reproducibility was less than 20% for both matrices. The expanded measurement uncertainty for the MCPD and glycidyl esters of the oil and powdered infant formula was less than 50%. For all parameters, the requirements set by the Customs Laboratory and the performance requirements of Regulation (EU) 1881/2006 were met. A method validated for two matrices can then be accredited. The customs laboratory may use the developed method in the future to control 3-MCPD, 2-MCPD and glycidyl esters levels of oils and powdered infant formulas. In the future, the method could also be validated for new matrices, such as liquid infant formulas.

Lactic acid bacteria have a long history of use in food industry due to their favorable metabolic properties and health benefits for human health. Therefore, they are generally recognized as safe (GRAS) by FDA (U.S Food and Drug Administration) and have QPS (Qualified Presumption of Safety) status granted by EFSA (European Food Safety Authority). Nowadays, antimicrobial resistance (AMR) is a serious global risk and due to the increasing AMRs, more and more microbial infections have become more difficult to treat with antibiotics. AMR has mainly been of concern in relation to pathogenic microbes. However, since fermented foods are favorable environments for AMR gene transfer it should also be considered in the context of beneficial bacteria and their potential to spread AMR genes into pathogenic microbes. The aim of this study was to determine antibiotic susceptibilities of Lactobacillus plantarum, Lactobacillus rhamnosus, Leuconostoc sp. and Weissella sp. strains by E-test method and to detect selected specific antibiotic resistance genes by PCR. In addition, the goal was to define new cut-off values for Weissella strains since, so far, these have not been defined by EFSA. Antibiotic susceptibilities were determined against eight antibiotics: ampicillin, chloramphenicol, clindamycin, erythromycin, gentamicin, kanamycin, streptomycin and tetracycline. The detected AMR genes were blaZ, mecA, cat, lnuA, tetK and tetM. Most of the determined strains were observed to exhibit a notable resistance to kanamycin. Several Leuconostoc sp. and L. rhamnosus strains showed also resistance to chloramphenicol. Interestingly, one L. rhamnosus strain was observed to exhibit multiresistance to chloramphenicol and clindamycin. Moreover, 48% Leuconostoc strains had higher MIC value for streptomycin than the cut-off value defined by EFSA. Any of the selected AMR genes were not detected even though a notable resistance during the phenotypic testing was observed. However, this might be explained by the small amount of detected AMR genes. The results obtained in the present study provided more information about the antibiotic susceptibility and the safety of L. plantarum, L. rhamnosus, Leuconostoc sp. and Weissella sp. strains. Moreover, new cut-off values were proposed for Weissella sp. strains.

Alzheimer’s disease (AD) is the seventh leading cause of death worldwide. One hallmark of AD includes the amyloid beta (Aβ1-42) peptide that accumulates into oligomers, fibrils, and plaques. Aβ1-42 has been shown to be structurally and functionally similar to antimicrobial peptides (AMPs). Publications have reported that Borrelia burgdorferi can be found in the brain of AD patients. B. burgdorferi and B. garinii cause Lyme disease (LD). B. duttonii is responsible for relapsing fever (RF), a disease characterized by recurrent episodes of high fever. The aim of this research was to study whether synthetic Aβ1-42 binds to LD and RF Borrelia sp. and several bacterial molecules important for their virulence, and whether Borrelia sp. have evolved strategies to evade Aβ1-42-mediated killing. Binding of Aβ1-42 to B. burgdorferi, B. garinii and B. duttonii and several microbial molecules was studied by ELISA and immunoblotting. Bacterial culturing and microscopy were used to study survival, agglutination, and phagocytosis of Borrelia sp. in the presence of Aβ1-42 and microglia. In this research, Aβ1-42 was able to bind and agglutinate all of the three studied Borrelia sp. However, Aβ1-42 reduced the survival and increased the phagocytosis of B. duttonii. while B. burgdorferi and B. garinii were unaffected. In addition, potential Aβ1-42 binding molecules were detected from several bacterial species, including FhbA expressed by B. duttonii. In conclusion, this study suggests that some restricted species of bacteria may evade Aβ1-42 entrapment and thus may be involved in the ability of the species to invade the CNS that may trigger neuroinflammation related to AD.

In recent years, consumer interest for probiotic products has significantly grown due to their health benefits, however challenges regarding viability and controlled release of probiotic bacteria during their processing and storage still exist. Microencapsulation of probiotics by spray drying with a suitable protective material could alleviate these challenges. This research aims to assess the viability of utilizing wood hemicelluloses recovered from forest industry side-streams, specifically galactoglucomannans (GGM) and glucuronoxylans (GX), as protective agents for the probiotic strain Lacticaseibacillus rhamnosus GG (LGG) during spray drying. The study results were compared to those of maltodextrins, considering varying solid concentrations (15 and 20%) and inlet air drying temperatures (105 and 140 oC). Feed dispersion properties including viscosity, pH, particle diameter and physical stability were determined. The probiotic's viability pre- and post-spray drying was evaluated, alongside powder characterization for moisture content, water activity, particle size, morphology and structure. Results indicated that both GGM and GX effectively shield LGG from heat impact during spray drying, yielding microcapsule powders with desirable attributes such as amorphous structure and low water activity. High encapsulation efficiency (>90%) comparable to maltodextrins suggested hemicelluloses as sustainable alternatives for conventionally used wall materials. Inlet air temperature or solid concentration did not affect encapsulation efficiency of hemicelluloses. Probiotic counts met a recommended level for probiotic products, signifying potential applications in food and pharmaceuticals. Powder yield, which varied between 35 and 58%, was significantly influenced by the encapsulating material. Morphological studies demonstrated well-formed, spherical particles at a specific drying temperature. The study proposes the potential use of wood hemicelluloses for effective probiotic protection, offering new possibilities for synbiotic powder applications in diverse industries due to their prebiotic properties which have been well reported in literature. Despite promising results, long-term stability and process optimization to improve the process yield and achieve lower moisture content for the microcapsule powders require further investigation. Adjusting feed dispersion parameters and exploring varied concentrations of hemicelluloses could enhance product yield. Meanwhile, increasing outlet air temperature possibly reduces the moisture content. This research fosters sustainable development in the forest industry, presenting a novel avenue for natural functional ingredient production.

Aspergillus niger is a filamentous fungus that is known for its ability to degrade plant biomass polysaccharides. A total of 86 sugar transporters have been identified in A. niger, but only 10 of them have been thoroughly characterized. Sugar transporter proteins are crucial for fungi as they enable efficient utilization of sugars in their metabolism and therefore breakdown of plant biomass. Additionally, sugar transporters can be used in various biotechnological applications. L-arabinose is a pentose sugar present in plant biomass and A. niger can utilize it through the pentose catabolic pathway (PCP). Recently, a sugar transporter LatA was identified from A. niger, capable of transporting the PCP intermediate product L-arabitol into fungal cells. L-arabitol is a polyol similar to xylitol and can be used as a low-calorie sweetener in food and beverage industries. Although A. niger LatA has previously been shown to be specific to L-arabitol in vivo, its in vitro functional activity has not yet been described. This study aimed to in vitro characterize two potential L-arabitol transporters from A. niger, LatA and unpublished 9364, using the yeast Saccharomyces cerevisiae. As a platform strain, we used S. cerevisiae IMK1010 that is devoid of all hexose and disaccharide transporters, as well as disaccharide hydrolases. In addition, we used a disaccharide-polyol and a pentose metabolic strain which were generated from the IMK1010 strain. The metabolic strains carried pathways for maltose, saccharose, sorbitol and mannitol, and xylose and arabinose, respectively. This provided a controlled research environment for studying A. niger LatA and 9364 transporters. The sugar specificity of the transporters was tested through two different growth experiments on solid media with all the strains and in liquid media with IMK1010 strains. The tested sugars included D-glucose, D-fructose and D-mannose hexoses, D-xylose and L-arabinose pentoses, maltose and sucrose disaccharides, and D-mannitol and D-sorbitol polyols. In addition, LatA was examined through a disappearance assay, measuring the loss of sugar from the liquid growth medium. Altogether four different combination gene constructs, green fluorescent protein (GFP) gene fusions and plain sugar transporter gene constructs were successfully engineered and 21 different transformant yeast strains produced for this study. GFP gene fusions, were in addition to growth experiments, used to study the localization of the sugar transporters to the cell membrane. In strains containing combination gene constructs encoding sugar transporters and GFP, the sugar transporters were successfully localized to the cell membrane, showing already that the transporters potentially have transport activity in the heterologous expression system. Based on the results, A. niger 9364 transported the tested hexoses and maltose in the growth experiments but did not transport tested pentoses, disaccharides or D-mannitol and D-sorbitol polyols. As expected, A. niger LatA did not transport any of the tested sugars, confirming its specificity to L-arabitol polyol. However, in the disappearance assay LatA unexpectedly did not transport L-arabitol. This might be due to the possible toxicity of the polyols in high concentrations to yeast cells and many of them also serve as regulators of osmotic pressure in cells, which may lower the transport capacity of the sugar transporters. In the future the function of the transporters can be tested in different sugar concentrations and pH in disappearance assay. Alternatively, a L-arabitol metabolic strain could be constructed to investigate sugar specificity using the growth experiment instead of the disappearance assay. The study provided new information of A. niger 9364 and further insights into the sugar specificity of A. niger LatA. These sugar transporters could be used in various biotechnological applications in the future.

Infants´ and children´s diet influences normal development and overall health. A balanced diet providing essential nutrients is crucial. Recent research has examined the dietary patterns of children and infants, exploring potential associations between food components and the emergence of illnesses. Notably, investigations into relationships between dietary factors and metabolite have gained prominence. Metabolomics offers a means to investigate individual´s nutrition, health status, illnesses, and the interaction of medications and contaminants. This study aimed to elucidate the connections between diet and the serum metabolic profiles of 1-year-old Finnish children. This master´s thesis used data from Finnish infants (n=439) collected by 3-day food record and questionnaires, in conjunction with metabolite assessments from blood samples collected at the age of 12 months. The investigation particularly focused on cow´s milk products and breast milk. Spearman correlation coefficient served as the primary statistical tool utilising data derived from the DIPP Nutrition study. Infant diets´ primarily comprised various cow´s milk products, milks and infant formulas. Noteworthy findings revealed that distinct lipids and free fatty acids, significantly associated with cow´s milk product consumption and breastfeeding. In the future, this study holds potential for enhancing comprehension of diet-related disease development by employing metabolites as markers to dissect dietary impacts.

Tämän tutkielman tavoitteena oli tuottaa Propionibacterium freduenreichii -bakteerin ja Rhizopus -homesienen yhteisfermentaatiolla B12-vitamiinia härkäpavusta valmistettuun tempeen. Samalla tutkittiin, mitä aminohappoja ja orgaanisia happoja tempeissä vapautui fermentaation aikana ja vaikuttivatko ne B12-vitamiinin tuottoon. B12-vitamiinia sisältävien kasviperäisten elintarvikkeiden tuottamisen tutkiminen on tärkeää, sillä se helpottaisi vegaanista ruokavaliota noudattavien mahdollisuuksia saada B12-vitamiinia ravinnostaan. Hypoteesina oli, että B12-vitamiinia saataisiin härkäpaputempeihin, mutta oletettavasti sen pitoisuuteen valmiissa tempeissä vaikuttaisi käytetty propionibakteerikanta. Ensin kasvatettiin fermentointiin käytetyt propionibakteerikannat ja valmistettiin tempenäytteet härkäpavusta. Valmiista tempeistä määritettiin pH, kosteuspitoisuus, B12-vitamiini, vapaat aminohapot, orgaaniset hapot ja ammoniakki sekä propionibakteerien ja enterobakteerien määrät. Neljästä propionibakteerikannasta kaksi tuottivat tempeen huomattavia määriä B12-vitamiinia (1 µg/g kuivapainoa kohti). Fermentaation aikana vapaiden aminohappojen pitoisuus lisääntyi kaikilla kannoilla valmistetuissa tempeissä selvästi. Eri aminohappoja vapautui kuitenkin eri kannoilla hyvin eri määriä. Maitohappo kului näytteistä fermentoinnin aikana. Pelkällä Rhizopus -homesienellä valmistetussa näytteessä maitohapon määrä taas lisääntyi. Tämä oli tiedettävästi ensimmäinen kerta, kun tutkittiin härkäpaputempen valmistusta Rhizopus -homesienen ja propionibakteerin yhteisfermentaatiolla B12-vitamiinin tuottamiseksi. Tutkimus antoi lupaavia tuloksia B12-vitamiinipitoisen tempen valmistuksesta, sillä härkäpaputempejen vitamiinipitoisuudet olivat huomattavasti suurempia kuin mitä aikaisemmissa tutkimuksissa soija- ja lupiinitempeillä homesienen ja propionibakteerin yhteisfermentaatiossa saadut vitamiinipitoisuudet.