To diagnose patients suffering from blood clotting disorders latex immununoassays (LIA) can be used. A time consuming manual tangential flow filtration (TFF) process suggests the implementation of an automatic TFF system to improve the efficiency, profitability, and expandability of the production facility of LIA at Nordic Biomarker. Tests were made of the automatic TFF system's ability to perform the desired steps of concentration, dilution and diafiltration, both with purified water and mimicked product. The mimicked product of micro particles (MP) mixed with monoclonal antibodies (mAb) was also used to further test the system's pressure control, safety alarms and stops, and to determine a permeate flux by a critical flux experiment. The results imply a functional TFF system able to automatically concentrate the process fluid and maintain a stable volume during diafiltration, although an additional permeate pump was ordered to be able to attain a fully functional performance of the automatic TFF process. The final part of the implementation was to initiate a validation draft including a risk assessment, OQ plan and PQ plan that resulted in a plan of the main tests to be performed. To conclude, the essential part of the implementation of a high quality and efficient automatic TFF process was conducted to facilitate future expansion of the production of LIA.

Under covid-19-pandemin har avloppsvattenbaserad epidemiologi (WBE) använts i stor utsträckning som ett komplement till kliniska tester över många delar av världen. Detta projekt syftade till att detektera och kvantifiera belastningen av Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) i avloppsvattenprover med hjälp av Revers transkriptas kvantitativ polymeraskedjereaktion (RT-qPCR). De analyserade proverna kom från fyra olika avloppsreningsverk i Sverige, under perioden november 2022 till maj 2023. Studien omfattade en översikt över olika provtagnings- och analytiska tekniker och normaliseringsmetoder som används i WBE-studier, vilket betonade vikten av metodval. SARS-CoV-2-RNA upptäcktes i alla analyserade prover och infektionstrender kunde identifieras effektivt, inklusive COVID-19-vågen som observerades under semesterperioden. De dominerande varianterna som upptäcktes under denna övervakningsperiod var omikron variantens undergrupper, BA.2. och BA.2.75. Den veckovisa kvantifierade SARS-CoV-2-belastningen i avloppsvattenproverna visade en signifikant positiv korrelation till de kliniska fall som rapporterats i motsvarande avrinningsområden. Denna associering förstärktes ytterligare genom att normalisera SARS-CoV-2-innehållet med fekal biomarkör peppar milt fläckvirus (PMMoV). Dessutom har två metoder för tidig varning, nämligen medelvärdet plus två standardavvikelser (MSD) och positiv procentuell förändring (PPC), implementerats på avloppsvattendata, vilket pekar på vikten av att tillämpa sådana varningsmetoder för att ge förståeliga och tolkbara resultat. Denna studie ger värdefulla insikter om övervakning och analys av SARS-CoV-2 i avloppsvatten, vilket bidrar till utvecklingen av robusta system för tidig varning och folkhälsostrategier. / During the COVID-19 pandemic, wastewater-based epidemiology (WBE) has been applied extensively as a complementary tool to clinical testing across many parts of the globe. This project aimed to detect and measure the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) load in wastewater samples using Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). The analyzed samples were from four different wastewater treatment plants (WWTPs) in Sweden, covering the period from November 2022 through May 2023. The study encompassed an overview of various sampling and analytical techniques and normalization approaches employed in WBE studies, highlighting the importance of method selection. SARS-CoV-2 RNA was detected in all the samples analyzed, and infection trends could be identified effectively, including the COVID-19 peak observed during the holiday season. The dominant variants detected during this monitoring period were the omicron variants; omicron BA.2. and omicron BA.2.75. The weekly quantified SARS-CoV-2 load in the wastewater samples showed a significant positive correlation to the clinical cases reported in the corresponding catchment areas. This association was further enhanced by normalizing SARS-CoV-2 content with the fecal biomarker pepper mild mottle virus (PMMoV). Furthermore, two early warning methods, namely the mean plus two standard deviations (MSD) and positive percentage change (PPC), were implemented on the wastewater data pinpointing the importance of applying such warning methods to provide understandable and interpretable results. This study provides valuable insights into the monitoring and analysis of SARS-CoV-2 in wastewater, contributing to the development of robust early warning systems and public health strategies.

Consumption of citrus fruits and citrus juice production creates wastes, which could be valorized by using it for cultivating fungi. Before cultivation, the medium needs to be sterilized though autoclavation. Larger volumes used when autoclaving requires longer heating cycles and therefore runs the risk of degrading the medium to a greater extent. This research examines the effects of the volume lemon waste medium used while sterilizing. The aim is to find the largest volume still providing good growth for the filamentous fungus used, Rhizopus Delemar. Lemon waste was provided by Herrljunga Musteri AB and was pre-treated at 45°C for 2h. The liquid was strained and autoclaved in different volumetric series ranging from 200 – 10 000 mL, that was then used in 200 mL shake flask cultivations.  A scale up in two 3,5 L bubble column reactors was also performed from the 10 000 mL autoclaved medium, after not observing severe impacts on growth. Testing was done by weighing biomass and HPLC analysis of sugars. The yield of the biomass in the shake flasks ranged from 0,11 – 0,14 g/g sugars and the biomass concentration ranged between 2,4 - 3,0 g/L. Overall, the volume of autoclavation seems to not too be of great concern when cultivating R. Delemar on lemon waste medium in the analyzed ranges.

I detta projekt har uttrycket av Affibodymolekyler i bioreaktorer av olika volymetriska skalor jämförts för att fastställa om ett tillförlitligt samband mellan de olika bioreaktorernas prestanda kan etableras för att möjliggöra utvecklingen av en nedskalad produktionsmodell för Affibodymolekyler. Baslinjen för jämförelsen i denna studie har varit en enliters- stirred tank reactor (STR) som de andra (mindre) bioreaktorernas prestanda jämfördes med. Jämförelsen av prestanda gjordes genom odling och uttryck av sex olika Affibodymolekyler i replikat i varje bioreaktorstorlek. Prestandan i detta fall hänvisar till produktionen av Affibodymolekyler (mg/L Cellodling) under odlingen, som fastställdes genom ett protokoll för proteinrening av lösligt intracellulärt protein genom affinitetskromatografi och kvantifiering genom absorbans vid 280 nm. De sex olika Affibodymolekyler som har studerats i detta projekt hade tidigare visat sig ha olika uttrycksnivåer, och har här jämförts med varandra i de olika bioreaktorskalorna. De två olika bioreaktorstorlekarna som bedömdes var en 300 mL skakkolv med 50 mL arbetsvolym och en mikrotiterplatta (MTP) med 3 mL arbetsvolym. Dessutom innebar studien en bedömning av två system för en långsam frisättning av kolkälla i odlingarna, ett i varje nedskalad bioreaktorstorlek. Detta utfördes för att delvis efterlikna den kontrollerade fed-batch-kulturen i STR, där koltillförsel kontrollerades med hjälp av en feedprofil. Resultaten visade att proteinuttrycket av metoderna med en långsam frisättning av kolkällor överensstämde närmast med proteinuttrycket i STR. Anpassningen av dessa resultat mot proteinuttrycket hos STR gav i en linjär regression ett R2 på 99,69 % i 3 ml MTP och 97,46 % i 50 ml skakkolven. Slutsatserna som drogs var att SMFP08003 FeedPlate var den bästa kandidaten för en nedskalad modellering av uppströmsprocessen för produktion av Affibodymolekyler. Samt att faktorn att använda en fed-batch-process istället för en batch-process har en större inverkan på proteinuttrycket än skalan av processerna. / In this project the expression of Affibody® molecules in bioreactors of different volumetric scales has been compared, to determine if a reliable relation between the performance of the different bioreactors can be established to allow for the development of a scale-down Affibody® molecule production protocol. The baseline of comparison in this study has been a one litre Stirred Tank Reactor (STR) to which the other (smaller) bioreactors' performance were compared. The performance comparison was achieved by the cultivation and subsequent expression of six different Affibody® molecules in replicates in each bioreactor size. Performance in this case refers to Affibody® molecule production (mg/L culture) during the cultivation, which is assessed by a protocol of protein purification of soluble intracellular protein by affinity chromatography and quantification by 280 nm absorbance. The six different Affibody® molecules studied in this project had previously been found to have different expression levels, and were in this project compared to each other in the different bioreactor scales. The two different bioreactor sizes which were assessed were a 300 mL shake flask with 50 mL working volume and a 3 mL working volume microtiter plate (MTP). In addition, the study involved an assessment of the use of two systems for a slow carbon source release in the cultivations, one in each scale-down bioreactor size. This was performed to partly mimic the controlled feed systems in STRs. The results showed that the protein expression of the methods with a slow carbon source release corresponded most closely with the protein expression in the STR. The fit of these results onto the protein expression of the STR yielded a R2 of 99.69% in the 3 mL MTP and 97.46% in the 50 mL shake flask. The conclusions drawn were that SMFP08003 FeedPlate was the best candidate for scale-down modelling of the upstream process for production of Affibody® Molecules and that the factor of using a fed-batch process instead of a batch process has a larger impact on the protein expression than the scale of either process.

Attention-deficit/hyperactivity disorder (ADHD), is a behaviour disorder, usually first diagnosed in childhood, that is characterized by inattention, impulsivity, and hyperactivity. ADHD is often associated with co-morbid disorders like bipolar disorder, anxiety, depression, and substance abuse. The diagnosis of ADHD is clinically established by a review of symptoms and impairment from the child’s young age. There are numerous assistive technologies that exist for people suffering from ADHD but there exists a research gap in developing self-reflective tools for people with neurodevelopmental disorders. This paper bridges this research gap for students at Malmö University. This project will focus on developing a personalized interactive AI-based system that captures contextual data, analyses it to find relevant patterns in user’s behaviour, and visualizes it effectively to provide students with ADHD with insights into the parameters influencing the nature of their disorder. The project is performed under a Double Diamond method which allows for iteration. The methods used mostly comprise co-design methods to ensure the concept caters to the user’s needs. The project is based on learnings from three key areas: Interactive AI, Personal Informatics and Systems as dialogue partners.

Syftet med projektet var att studera effekten av olika antimikrobiella produkter på bakterietillväxt. Resultatet visade att Micrococcus luteus inte hade någon aktivitet i det 10% tunndrank-baserade LB-mediet (specifikt odlingsmedium för bakterien där tunndrank, en biprodukt från etanolindustri, har legat vid 10% koncentration) som användes under experimentet, medan antimikrobiellen Fermasure i höga koncentrationer nämligen 1600 ppm var effektivt mot bakterien Enterococcus faecium. Antimikrobiellen Vitahop vid koncentration av 25 ppm, hade framgång mot tillväxten av bakterien. Dessa antimikrobiella produkter kan användas vid odlingen av svamp för att inhibera tillväxten av bakterier. Resultaten kan hjälpa till att fastställa doseringen av antimikrobiella produkter under svamptillväxt i stor skala där kontamination kan vara ett problem. / The aim of the project was to study the effect of various antimicrobial products on bacterial growth. The results showed that Micrococcus luteus had no activity in the 10% thin-rank-based LB medium (specifically culture medium for the bacterium where thin draff, a by-product of the ethanol industry, has been at 10% concentration) used during the experiment, while the antimicrobial Fermasure in high concentrations namely 1600 ppm was effective against the bacterium Enterococcus faecium. The antimicrobial Vitahop at concentration of 25 ppm, had success against the growth of the bacterium. These antimicrobial products can be used in the cultivation of fungus to inhibit the growth of bacteria. The findings could help determine the dosage of antimicrobial products during fungal growth on a large scale where contamination can be a problem.

Med nya utmaningar för att tillgodose behoven av tjänligt dricksvatten hos konsumenter över hela världen krävs innovativa tekniker för övervakning av vattenkvalitet. I det här projektet undersöktes ett nytt instrument som detekterar och klassificerar partiklar i dricksvatten med hjälp av maskininlärda modeller. Målet var att utvärdera dess användbarhet som ett onlinesystem för tidig varning på Norrvattens reningsverket och ledningsnät. Utvärderingen utfördes som två separata delar: (1) en översiktlig analys av data som tidigare samlats in av tre instrument placerade på Norrvattens reningsverk och ledningsnät med målet att hitta trender och definiera tröskelnivåer, och (2) genom att utföra spikningsexperiment i instrumentet med kända föroreningar i laboratoriemiljö. Föroreningarna som undersöktes var E. coli, B. megaterium, humussyror, cyanobakterier av stam Synechocystis PCC 6803, och biofilm. Flödescytometri genomfördes på samma föroreningar för att möjliggöra jämförelser. Dataanalysen visade att instrumentet kan upptäcka säsongsvariationer i partikelnivåer. Dessutom har det partikelklasser som inte varierade med dessa fluktuationer vilket gör dem lovande som oberoende parametrar i ett varningssystem. Det fanns dock indikationer på att instrumentet kan göra oförutsedda klassifikationer av partiklar utifrån skillnader i sammansättningen mellan träningsdatats vatten och vattnet på Norrvatten. De laborativa experimenten visade att instrumentet kunde detektera alla föroreningar som testades, även vid cellantal på några få hundra celler/mL. Det kunde jämföras med flödescytometern, där det nya instrumentet kunde upptäcka halter av cyanobakterier under detektionsgränsen för flödescytometern, vilket indikerar en hög känslighet. Därför drogs slutsatsen att instrumentet har potential som tidigt varningssystem, men dess användbarhet hos Norrvatten är begränsad i dess nuvarande tillstånd på grund av de oförutsedda klassificeringarna av partiklar i deras vatten. / With emerging challenges in ensuring safe supplies of drinking water to consumers worldwide, there is a need for innovative technologies for water quality monitor- ing. In this project, an instrument which can detect and classify particles in drinking water using machine-learned models was investigated. The aim was to assess its usefulness as an online, early warning system in the drinking water treatment plant and distribution system at Norrvatten. The assessment was conducted as two separate parts: (1) an analysis of data previously collected by three instruments located around Norrvatten’s plant and distribution system, with the aim of finding trends and creating baselines, and (2) by conducting spiking experiments in the instrument using known contaminants in a lab environment. The contaminants tested were E. coli, B. megaterium, humic acids, cyanobacteria Synechocystis PCC 6803, and biofilm. Flow cytometry was performed on the same contaminants to enable comparison. It could be concluded from the data analysis that the instrument can detect seasonal trends in particle levels. In addition, there are classes of particles which are not subject to these fluctuations, making them promising as independent parameters in a warning system. There were however indications that the instrument can make unexpected classifications of particles due to differences in composition in the training data water and the water at Norrvatten. The lab experiments showed that the instrument could detect all contaminants tested, even cell numbers of a few hundred cells/mL. Comparing with flow cytometry, the novel instrument could detect concentrations of cyanobacteria below the detection limit of the flow cytometer, indicating a high sensitivity. It was concluded that the instrument has properties desired in an early warning system, but its usefulness at Norrvatten is limited in its current state due to the unexpected classifications of particles in their water.

This thesis investigates a newly developed and patented technology for its ability to inactivate spore- forming bacteria and non-spore-forming microorganisms. This new technology “Pressure Assisted Thermal Sterilization©” (PATS) is based on the theory of the thermal expansion of liquids. The efficiency of inactivating spore-forming and non-spore-forming microorganisms by PATS was compared with the thermal treatment alone. A combination treatment consisting of high pressure processing and gaseous carbon dioxide was also investigated for its ability to inactivate bacterial spores in model and real food matrices. The structural damage caused by treatments to the spores and non-spore-forming bacteria was assessed by scanning electron microscopy. Geobacillus stearothermophilus spores suspended in Milli-Q water, UHT milk and pumpkin soup, treated by PATS were found to have significantly lower decimal reduction times (D values) compared with the thermal treatment alone. Spores suspended in UHT milk were more heat resistant compared with those in Milli-Q water and pumpkin soup. Bacillus cereus spores suspended in Milli-Q water and pumpkin soup treated with PATS were more effectively inactivated compared with spores treated by the thermal treatment alone. Clostridium botulinum spores in saline buffer subjected to PATS treatment were inactivated more effectively compared with the thermal treatment alone. Overall, the results show that PATS was a better processing technique for inactivation of bacterial spores compared with thermal treatment alone. However, PATS had no added benefit in inactivating the non-spore-forming bacteria Escherichia coli and Saccharomyces cerevisiae cells compared with the thermal treatment. A shelf life study showed that B. cereus spores in pumpkin soup retained a low spore count (<5 LogCFU/mL) for approximately 40 days in 30oC storage after treatment with PATS. No additional degradation of colour pigments of pumpkin soup and model pumpkin juice was observed following PATS compared with the thermal treatment. Spore-forming microorganisms can be resistant to pressure treatment alone, which limits the application of high pressure processing (HPP). Therefore, a combination approach was investigated. The mechanism of inactivating spores by combining HPP with other treatments is that the pressure assists in spore germination. Then a secondary treatment (thermal or CO2 gas) can be used to inactivate the germinated spores. A combined application of HPP and a consecutive CO2 treatment was investigated for the efficiency of spore inactivation. Results showed that HPP (200 MPa for 30 min) followed by a CO2 treatment inactivated Bacillus subtilis 168 in nutrient broth, tomato juice and liquid whole egg by 2.5, 1.0 and 1.5 LogCFU/mL respectively. These results indicated that this technique is inadequate for practical use. Scanning electron micrographs showed that pressure processing of B. subtilis 168 and B. subtilis natto spores resulted in deformation of the spore structure. This structural deformation of spores may have been due to water absorption during HPP and subsequent release upon decompression. PATS treated G. stearothermophilus and B. cereus spores were more severely damaged compared with the same spores which underwent thermal treatment alone. However, the extent to which E. coli and S. cerevisiae cells were damaged by both PATS and thermal treatment was similar.

This thesis investigates a newly developed and patented technology for its ability to inactivate spore- forming bacteria and non-spore-forming microorganisms. This new technology “Pressure Assisted Thermal Sterilization©” (PATS) is based on the theory of the thermal expansion of liquids. The efficiency of inactivating spore-forming and non-spore-forming microorganisms by PATS was compared with the thermal treatment alone. A combination treatment consisting of high pressure processing and gaseous carbon dioxide was also investigated for its ability to inactivate bacterial spores in model and real food matrices. The structural damage caused by treatments to the spores and non-spore-forming bacteria was assessed by scanning electron microscopy. Geobacillus stearothermophilus spores suspended in Milli-Q water, UHT milk and pumpkin soup, treated by PATS were found to have significantly lower decimal reduction times (D values) compared with the thermal treatment alone. Spores suspended in UHT milk were more heat resistant compared with those in Milli-Q water and pumpkin soup. Bacillus cereus spores suspended in Milli-Q water and pumpkin soup treated with PATS were more effectively inactivated compared with spores treated by the thermal treatment alone. Clostridium botulinum spores in saline buffer subjected to PATS treatment were inactivated more effectively compared with the thermal treatment alone. Overall, the results show that PATS was a better processing technique for inactivation of bacterial spores compared with thermal treatment alone. However, PATS had no added benefit in inactivating the non-spore-forming bacteria Escherichia coli and Saccharomyces cerevisiae cells compared with the thermal treatment. A shelf life study showed that B. cereus spores in pumpkin soup retained a low spore count (<5 LogCFU/mL) for approximately 40 days in 30oC storage after treatment with PATS. No additional degradation of colour pigments of pumpkin soup and model pumpkin juice was observed following PATS compared with the thermal treatment. Spore-forming microorganisms can be resistant to pressure treatment alone, which limits the application of high pressure processing (HPP). Therefore, a combination approach was investigated. The mechanism of inactivating spores by combining HPP with other treatments is that the pressure assists in spore germination. Then a secondary treatment (thermal or CO2 gas) can be used to inactivate the germinated spores. A combined application of HPP and a consecutive CO2 treatment was investigated for the efficiency of spore inactivation. Results showed that HPP (200 MPa for 30 min) followed by a CO2 treatment inactivated Bacillus subtilis 168 in nutrient broth, tomato juice and liquid whole egg by 2.5, 1.0 and 1.5 LogCFU/mL respectively. These results indicated that this technique is inadequate for practical use. Scanning electron micrographs showed that pressure processing of B. subtilis 168 and B. subtilis natto spores resulted in deformation of the spore structure. This structural deformation of spores may have been due to water absorption during HPP and subsequent release upon decompression. PATS treated G. stearothermophilus and B. cereus spores were more severely damaged compared with the same spores which underwent thermal treatment alone. However, the extent to which E. coli and S. cerevisiae cells were damaged by both PATS and thermal treatment was similar.

This thesis investigates a newly developed and patented technology for its ability to inactivate spore- forming bacteria and non-spore-forming microorganisms. This new technology “Pressure Assisted Thermal Sterilization©” (PATS) is based on the theory of the thermal expansion of liquids. The efficiency of inactivating spore-forming and non-spore-forming microorganisms by PATS was compared with the thermal treatment alone. A combination treatment consisting of high pressure processing and gaseous carbon dioxide was also investigated for its ability to inactivate bacterial spores in model and real food matrices. The structural damage caused by treatments to the spores and non-spore-forming bacteria was assessed by scanning electron microscopy. Geobacillus stearothermophilus spores suspended in Milli-Q water, UHT milk and pumpkin soup, treated by PATS were found to have significantly lower decimal reduction times (D values) compared with the thermal treatment alone. Spores suspended in UHT milk were more heat resistant compared with those in Milli-Q water and pumpkin soup. Bacillus cereus spores suspended in Milli-Q water and pumpkin soup treated with PATS were more effectively inactivated compared with spores treated by the thermal treatment alone. Clostridium botulinum spores in saline buffer subjected to PATS treatment were inactivated more effectively compared with the thermal treatment alone. Overall, the results show that PATS was a better processing technique for inactivation of bacterial spores compared with thermal treatment alone. However, PATS had no added benefit in inactivating the non-spore-forming bacteria Escherichia coli and Saccharomyces cerevisiae cells compared with the thermal treatment. A shelf life study showed that B. cereus spores in pumpkin soup retained a low spore count (<5 LogCFU/mL) for approximately 40 days in 30oC storage after treatment with PATS. No additional degradation of colour pigments of pumpkin soup and model pumpkin juice was observed following PATS compared with the thermal treatment. Spore-forming microorganisms can be resistant to pressure treatment alone, which limits the application of high pressure processing (HPP). Therefore, a combination approach was investigated. The mechanism of inactivating spores by combining HPP with other treatments is that the pressure assists in spore germination. Then a secondary treatment (thermal or CO2 gas) can be used to inactivate the germinated spores. A combined application of HPP and a consecutive CO2 treatment was investigated for the efficiency of spore inactivation. Results showed that HPP (200 MPa for 30 min) followed by a CO2 treatment inactivated Bacillus subtilis 168 in nutrient broth, tomato juice and liquid whole egg by 2.5, 1.0 and 1.5 LogCFU/mL respectively. These results indicated that this technique is inadequate for practical use. Scanning electron micrographs showed that pressure processing of B. subtilis 168 and B. subtilis natto spores resulted in deformation of the spore structure. This structural deformation of spores may have been due to water absorption during HPP and subsequent release upon decompression. PATS treated G. stearothermophilus and B. cereus spores were more severely damaged compared with the same spores which underwent thermal treatment alone. However, the extent to which E. coli and S. cerevisiae cells were damaged by both PATS and thermal treatment was similar.