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The physiological effects of oxygen on Limosilactobacillus reuteri DSM 17938 during cultivation and the ensuing properties of the freeze-dried probiotic product was investigated. On-line flow cytometry and k-means clustering gating was used to follow growth and viability in real time during cultivation. The bacterium tolerated aeration at 500 ml/min, with a growth rate of 0.74 ± 0.13 h-1 which demonstrated that low levels of oxygen did not influence the growth kinetics of the bacterium. Modulation of the redox metabolism was, however, seen already at non-inhibitory oxygen levels by 1.5-fold higher production of acetate and 1.5-fold lower ethanol production. A significantly higher survival rate in the freeze-dried product was observed for cells cultivated in presence of oxygen compared to absence of oxygen (61.8 ± 2.4 % vs 11.5 ± 4.3 %), coinciding with a higher degree of unsaturated fatty acids (UFA:SFA ratio of 10 for air sparged vs 3.59 for N2 sparged conditions.). Oxygen also resulted in improved bile tolerance and boosted 5’nucleotidase activity (370 U/L vs 240 U/L in N2 sparged conditions) but lower tolerance to acidic conditions compared bacteria grown under complete anaerobic conditions which survived up to 90 min of exposure at pH 2. Overall, our results indicate the controlled supply of oxygen during production may be used as means for probiotic activity optimisation of L. reuteri DSM 17938.

A Gram-stain-negative, rod-shaped strain, Braz8T, isolated from larvae ofAnopheles darlingiwas investigatedusing a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences showed that strainBraz8Twas related most closely to species of the genusThorsellia, with 95.6, 96.5 and 96.6% similarity to thetype strains ofThorsellia anophelis,Thorsellia kandunguensisandThorsellia kenyensis, respectively, andformed a separate branch in the phylogenetic tree next to the monophyletic cluster of the genusThorsellia.Chemotaxonomic data supported the allocation of the strain to the familyThorselliaceae. The major fatty acidswere C18:1!7c, C16:0and C14:0. The quinone system was composed of ubiquinones Q-8 and Q-7 (1:0.3),the predominant polar lipids were diphosphatidylglycerol and phosphatidylglycerol, and the polyamine patternshowed the major compound putrescine. However, qualitative and quantitative differences in the majorpolyamine, polar lipid profile and fatty acid patterns distinguished strain Braz8Tfrom species of the genusThorsellia. Phylogenetic analysis based on 16S rRNA gene sequences, average nucleotide identity, DNA–DNAhybridization, multilocus sequence analysis as well as physiological and biochemical tests distinguished strainBraz8Tboth genotypically and phenotypically from the threeThorselliaspecies but also showed its placementin the familyThorselliaceae. Thus, strain Braz8Tis considered to represent a novel species of a new genus mostclosely related to the genusThorsellia, for which the nameCoetzeea brasiliensisgen. nov., sp. nov. is proposed.The type strain ofCoetzeea brasiliensisis Braz8T(=LMG 29552T=CIP 111088T)

Two Gram-negative, rod-shaped strains, T2.1T and W5.1.1T, isolated from the larvae of the mosquito Anopheles arabiensis were investigated using a polyphasic approach. On the basis of 16S rRNA gene sequence similarity studies, strains T2.1T and W5.1.1T were shown to belong to the genus Thorsellia, both with 97.8% similarity to the type strain of T. anophelis, and with 98.1% similarity to each other. Chemotaxonomic data supported the allocation of the strains to the genus Thorsellia: their major fatty acids are C18:1ω7c, C16:0, and C14:0; they harbor a ubiquinone Q-8 quinone system and a polyamine pattern with the major compound 1,3-diaminopropane. Qualitative and quantitative differences in their polar lipid profile distinguish the two strains T2.1T and W5.1.1T from each other and from T. anophelis. Average nucleotide identity (ANI), DNA-DNA hybridization, Multi Locus sequence analysis (MLSA) as well as physiological and biochemical tests allowed to distinguish both genotypically and phenotypically T2.1T and W5.1.1T from each other and from the type strain of T. anophelis. Thus, we propose that these isolates represent two novel species of the genus Thorsellia, named Thorsellia kenyensis sp. nov. and Thorsellia kandunguensis sp. nov., respectively. The type strains are T2.1T (=CCM 8545 = LMG 28483T) and W5.1.1T (= LMG 28213T =CIP 110794T), respectively. Furthermore, phylogenetic analysis based on nearly full-length 16S rRNA gene sequences showed that the genus Thorsellia formed a separate branch, distinct from Enterobacteriaceae, Pasteurellaceae, and Orbaceae. As a consequence, a new family Thorselliaceae fam. nov. is proposed.

Cellular water can be removed to reversibly inactivate microorganisms to facilitate storage. One such method of removal is freeze-drying, which is considered a gentle dehydration method. To facilitate cell survival during drying, the cells are often formulated beforehand. The formulation forms a matrix that embeds the cells and protects them from various harmful stresses imposed on the cells during freezing and drying. We present here a general method to evaluate the survival rate of cells after freeze-drying and we illustrate it by comparing the results obtained with four different formulations: the disaccharide sucrose, the sucrose derived polymer Ficoll PM400, and the respective polysaccharides hydroxyethyl cellulose (HEC) and hydroxypropyl methyl cellulose (HPMC), on two strains of bacteria, P. putida KT2440 and A. chlorophenolicus A6. In this work we illustrate how to prepare formulations for freeze-drying and how to investigate the mechanisms of cell survival after rehydration by characterizing the formulation using of differential scanning calorimetry (DSC), surface tension measurements, X-ray analysis, and electron microscopy and relating those data to survival rates. The polymers were chosen to get a monomeric structure of the respective polysaccharide resembling sucrose to a varying degrees. Using this method setup we showed that polymers can support cell survival as effectively as disaccharides if certain physical properties of the formulation are controlled.

Freeze-drying of bacterial cells with retained viability and activity after storage requires appropriate formulation, i.e. mixing of physiologically adapted cell populations with suitable protective agents, and control of the freeze-drying process. Product manufacturing may alter the clinical effects of probiotics and it is essential to identify and understand possible factor co-dependencies during manufacturing. The physical solid-state behavior of the formulation and the freeze-drying parameters are critical for bacterial survival and thus process optimization is important, independent of strain. However, the maximum yield achievable is also strain-specific and strain survival is governed by e.g. medium, cell type, physiological state, excipients used, and process. The use of preferred compatible solutes for cross-protection of Lactobacilli during industrial manufacturing may be a natural step to introduce robustness, but knowledge is lacking on how compatible solutes, such as betaine, influence formulation properties and cell survival. This study characterized betaine formulations, with and without sucrose, and tested these with the model lactic acid bacteria Lactobacillus coryniformis Si3. Betaine alone did not act as a lyo-protectant and thus betaine import prior to freeze-drying should be avoided. Differences in protective agents were analyzed by calorimetry, which proved to be a suitable tool for evaluating the characteristics of the freeze-dried end products.

Background: Disaccharides are in general first choice as formulation compounds when freezedrying microorganisms. Although polysaccharides and other biopolymers are considered too large to stabilize and interact with cell components in the same beneficial way as disaccharides, polymers have been reported to support cell survival. In the present study we compare the efficiency of sucrose, the polymers Ficoll, hydroxyethylcellulose, hydroxypropylmethylcellulose and polyvinylalcohol to support survival of three bacterial strains during freeze-drying. The initial osmotic conditions were adjusted to be similar for all formulations. Formulation characterization was used to interpret the impact that different compound properties had on cell survival. Results: Despite differences in molecular size, both sucrose and the sucrose based polymer Ficoll supported cell survival after freeze-drying equally well. All formulations became amorphous upon dehydration. Scanning electron microscopy and X-ray diffraction data showed that the discerned differences in structure of the dry formulations had little impact on the survival rates. The capability of the polymers to support cell survival correlated with the surface activity of the polymers in a similar way for all investigated bacterial strains. Conclusion: Polymer-based formulations can support cell survival as effectively as disaccharides if formulation properties of importance for maintaining cell viability are identified and controlled.

The company is one of the largest in the world within its industry, producing and distributing its products in a global supply chain. The purpose of the project was to explore the potential of using Radio Frequency Identification (RFID) technology in the company global supply chain. The thesis students were working as project managers responsible for designing and implementing an RFID pilot in the company supply chain. The thesis concludes that RFID is a powerful technology that eventually will increase supply-chain visibility. However, there are several barriers to overcome before RFID will provide the operational benefits that the company are looking for, but also a number of distinct drivers of adoption of RFID in the supply-chain. Hence, there is a need for a structured approach to monitoring the development of RFID. The authors also present a framework for strategy under uncertainty enabling the company to stay up-to-date on the RFID development.

Venture capital-bolag tjänar sina pengar genom att investera i företags tidiga utvecklingsskeden, för att senare sälja sin del för en större summa än de investerade. Detta gör att tillfället då venture capital-bolaget säljer är väldigt kritiskt för hur mycket de skall tjäna på sin investering. I detta skede spelar självklart en mängd faktorer in på vilket pris som erhålls. Enligt samtal med flera venture capital-bolag har vi identifierat en faktor som verkar ge extra stora problem; huruvida de skulle genomföra en produktlansering av företagets produkt före de säljer eller om venture capital-bolaget skulle vänta med lanseringen och sälja först. Syftet med uppsatsen är att undersöka hur venture capital-bolags exit via trade sale påverkas av produktlansering i portföljföretag och därigenom ge svar på om venture capital företag ska göra exit före eller efter att portföljföretagets produkt lanserats. Uppsatsen är baserad på en kvalitativ undersökning av tre venture capital-bolag och sex portföljbolagsinvesteringar. En abduktiv ansats har använts som grund för arbetsprocessen och empiri är insamlad via upprepade öppna intervjuer. Den teoretiska referensramen utgörs av produktlanseringsteori, värderingsteori och beslutsteori. Slutsatsen utifrån den teori och empiri som behandlats är att en lyckad produktlansering i ett portföretag kan innebära att värdet på portföljföretaget minskar istället för det triviala - att värdet skulle öka. En produktlansering bör därför ej göras före trade sale. Venture capital-bolags möjligheter att påverka relationen produktlansering-trade sale är dessutom begränsade, mycket beroende på brist av köpare av portföljbolag innan genomförd produktlansering.

Syftet med uppsatsen var att utifrån ett projektarbete vid Malmö-Sturup Airport, lyfta fram och dra lärdom av grupprocesser och projektarbete. Viktiga lärdomar är att projektarbete ibland kräver förkunskaper inom process och metod snarare än branschspecifika kompetenser. En projektgrupp kan ha mer än en roll i arbetet och behöver då balansera dessa roller och relationer. En grupp med kompletterande personligheter når lättare effektivitet samtidigt som behovet av koordinator är stort. Styrningsverktyg är därför viktiga för att skapa struktur. En kaosartad problemformulering kan ge utrymme för stor kreativitet men ställer samtidigt högre krav på strukturerat arbete.